Home

Weiteres zum Thema:

Medizinische Chemie

Molekulare Medizin

Buecher zum Thema

Themenbezogene Artikel

- Klinische Chemie

- Krebsforschung

- Medizinische Chemie

- Pharmazeutische Chemie

- Proteomik

Weitere Themenbereiche

English Version

Verwandte Themen:

Biochemie

Aktuelles

Chemie Nachrichten

Stellenmarkt Chemie

Chemie Konferenzen

Chemie A bis Z

Index Chemie

Chemikalien

Internetchemie Lexikon

Produkte und Firmen

About Internetchemie

Internetchemie

Impressum

Service

Molekulare Medizin - Neueste Forschungsartikel

 
Aktuelle Fachartikel zur Molekularen Medizin, sortiert nach Erscheinungsdatum.

Die Urheberrechte und Veroeffentlichungsrechte der in der nachfolgenden Liste aufgefuehrten Fachartikel liegen bei den jeweiligen Verlagen, die am Ende des jeweiligen Artikels als Quelle genannt werden. Diese sind auch fuer die Inhalte verantwortlich.

Hinweise zur Veroeffentlichung Ihrer Pressmitteilung unter Internetchemie.Info entnehmen Sie bitte der entsprechenden Info-Seite.

Diese Seite koennen Sie mit folgender Tastenkombination nach Stichwoertern durchsuchen: <STRG> und <F>.

Auf dieser Seite beruecksichtige naturwissenschaftliche Journale:

Aktuelle wissenschaftliche Fachartikel der genannten Journale:

Distinct overlapping sequences at the carboxy-terminus of merlin regulate its tumor suppressor and morphogenic activity Abstract The Neurofibromatosis 2 (NF2) gene product merlin is a tumor suppressor, which in addition to inhibiting cell proliferation regulates cell morphology. The morphogenic properties of merlin may play a role in tumor suppression, since patient-derived tumor cells demonstrate cytoskeletal abnormalities. However, it is still unclear how these functions are linked. The N-terminal FERM-domain of merlin is highly homologous to the oncogenic protein ezrin, while the C-termini are less conserved, suggesting that the opposite effect of the proteins on proliferation could be mediated by their distinct C-terminal regions. In this study we characterize the role of the most C-terminal residues of merlin in the regulation of proliferation, cytoskeletal organization, phosphorylation, and intramolecular associations. In addition to the two full-length merlin isoforms and truncating mutations found in patients, we focused on the evolutionally conserved C-terminal residues 545-547, also harboring disease causing mutations. We demonstrate that merlin induces cell extensions, which result from impaired retraction of protrusions rather than from increased formation of filopodia. The residues 538-568 were found particularly important for this morphogenic activity. The results further show that both merlin isoforms are able to equally inhibit proliferation, whereas C-terminal mutants affecting residues 545-547 are less effective in growth suppression. This study demonstrates that the C-terminus contains distinct but overlapping functional domains important for regulation of the morphogenic activity, intramolecular associations, and cell proliferation. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 10 Feb 2012 | 7:33 pm CET

New Paper in Press HMGB1 Impairs P. aeruginosa Clearance in CF Lungs Quelle: Molecular Medicine | 8 Feb 2012 | 4:57 pm CET

Genetic variants implicated in telomere length associated with left ventricular function in patients with hypertension and cardiac organ damage Abstract   Telomere length has emerged as a biological correlate for ageing, which in turn is a risk factor for the manifestation of cardiovascular diseases. This study investigated the relation between leucocyte telomere length (LTL) and its genetic background to cardiac structure and function in patients with arterial hypertension. We analysed a cohort of 1,106 treated hypertensive patients (83.3% males; mean age, 57.9 ± 9.8 years) with an ejection fraction (EF) over 40% and documented cardiovascular disease or target organ damage. LTL and genotypes of single nucleotide polymorphisms (SNPs), previously implicated in LTL, were determined by real-time PCR. The mean left ventricular mass index (LVMI) and EF were 51.8 ± 21.0 g/H2.7 and 61.1 ± 9.6%, respectively. In multivariate adjusted analysis, a 1.5-fold LTL was positively related with a 2.2% increase of LVMI (CI = 0.1% to 4.2%, p = 0.044) and an absolute increase in EF of 0.6% (CI = 0.1% to 1.1%, p = 0.028). One SNP near TERC (rs16847897) showed a significant absolute difference in EF dependent on allele status (rs16847897, G allele 2.7%; CI = 0.7% to 4.6%; p raw = 0.008, p mt = 0.048, after adjustment for multiple testing). This applied also for two SNPs in BICD1 (rs2630578, C allele −1.8%; CI = −2.8% to −0.7%; p raw = 0.002, p mt = 0.018; rs1151026, G allele −1.9%, CI = −3.0% to −0.8%; p raw < 0.001, p mt = 0.002) with the extension that a frequent haplotype in BICD1 showed an absolute −1.8% (CI = −3.0% to −0.7%; p raw = 0.002, p mt = 0.008) lower EF compared with those lacking this haplotype. Our results point to a role of genetic variants recently implicated in LTL for left ventricular function in hypertensive patients. Content Type Journal Article Category Original Article Pages 1-9 DOI 10.1007/s00109-012-0874-3 Authors Matthias Huber, Institute of Clinical Pharmacology and Toxicology, CharitéCentrum für Therapieforschung, Charité–Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany Andras Treszl, Department of Medical Biometry and Epidemiology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany Markus Wehland, Institute of Clinical Pharmacology and Toxicology, CharitéCentrum für Therapieforschung, Charité–Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany Ingke Winther, Institute of Clinical Pharmacology and Toxicology, CharitéCentrum für Therapieforschung, Charité–Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany Irina Zergibel, Institute of Clinical Pharmacology and Toxicology, CharitéCentrum für Therapieforschung, Charité–Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany Rona Reibis, Institute of Clinical Pharmacology and Toxicology, CharitéCentrum für Therapieforschung, Charité–Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany Juliane Bolbrinker, Institute of Clinical Pharmacology and Toxicology, CharitéCentrum für Therapieforschung, Charité–Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany Monika Stoll, Leibniz-Institute for Arteriosclerosis Research, University of Münster, Münster, Germany Gilbert Schönfelder, Institute of Clinical Pharmacology and Toxicology, CharitéCentrum für Therapieforschung, Charité–Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany Karl Wegscheider, Department of Medical Biometry and Epidemiology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany Heinz Völler, Institute of Clinical Pharmacology and Toxicology, CharitéCentrum für Therapieforschung, Charité–Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany Reinhold Kreutz, Institute of Clinical Pharmacology and Toxicology, CharitéCentrum für Therapieforschung, Charité–Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 7 Feb 2012 | 7:08 pm CET

Role of CD95 in pulmonary inflammation and infection in cystic fibrosis Abstract   Cystic fibrosis is caused by a defective expression of the cystic fibrosis transmembrane conductance regulator (Cftr) gene, which results in chronic pulmonary inflammation and infections. The pathophysiological mechanisms by which these changes are induced in the lungs of patients with cystic fibrosis require definition. This study found that Cftr deficiency in mice results in the upregulation and activation of CD95. CD95 activation is caused by increased ceramide concentrations in cystic fibrosis lungs, as revealed by genetic modifications that normalize pulmonary ceramide concentrations. The activation of CD95 in cystic fibrosis lungs further increases pulmonary ceramide levels and results in a vicious feedback cycle of CD95 activation and ceramide accumulation. Genetic studies reveal that CD95 is crucially involved in the induction of aseptic inflammation, an increase in the bronchial cell death rate, and an increased susceptibility to infection of Cftr-deficient mice. All of these pathologies are partially corrected by heterozygosity of CD95 in Cftr-deficient mice. These findings identify CD95 as an important regulator of lung functions in cystic fibrosis and suggest that CD95 may be a novel target for treating cystic fibrosis. Content Type Journal Article Category Original Article Pages 1-13 DOI 10.1007/s00109-012-0867-2 Authors Katrin Anne Becker, Department of Molecular Biology, University of Duisburg-Essen, Hufelandstrasse 55, 45122 Essen, Germany Brian Henry, Department of Molecular Biology, University of Duisburg-Essen, Hufelandstrasse 55, 45122 Essen, Germany Regan Ziobro, Department of Molecular Biology, University of Duisburg-Essen, Hufelandstrasse 55, 45122 Essen, Germany Burkhard Tümmler, Clinical Research Group, OE6710, and Pediatric Pulmonology, Allergology and Neonatology, Department of Pediatrics and Adolescent Medicine, Hannover Medical School, Hannover, Germany Erich Gulbins, Department of Molecular Biology, University of Duisburg-Essen, Hufelandstrasse 55, 45122 Essen, Germany Heike Grassmé, Department of Molecular Biology, University of Duisburg-Essen, Hufelandstrasse 55, 45122 Essen, Germany Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 7 Feb 2012 | 7:08 pm CET

Hydrogen sulfide: a gasotransmitter of clinical relevance Abstract   Though the existence of hydrogen sulfide (H2S) in biological tissues has been known for over 300 years, it is the most recently appreciated of the gasotransmitters as a physiologic messenger molecule. The enzymes cystathionine γ-lyase (CSE) and cystathionine β-synthase (CBS) had long been speculated to generate H2S, and inhibitors of these enzymes had been employed to characterize influences of H2S in various organs. Definitive evidence that H2S is a physiologic regulator came with the development of mice with targeted deletion of CSE and CBS. Best characterized is the role of H2S, formed by CSE, as an endothelial derived relaxing factor that normally regulates blood pressure by acting through ATP-sensitive potassium channels. H2S participates in various phases of the inflammatory process, predominantly exerting anti-inflammatory actions. Currently, the most advanced efforts to develop therapeutic agents involve the combination of H2S donors with non-steroidal anti-inflammatory drugs (NSAIDs). The H2S releasing moiety provides cytoprotection to gastric mucosa normally adversely affected by NSAIDs while the combination of H2S and inhibition of prostaglandin synthesis may afford synergistic anti-inflammatory influences. Content Type Journal Article Category Review Pages 1-9 DOI 10.1007/s00109-012-0873-4 Authors M. Scott Vandiver, The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA Solomon H. Snyder, The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 7 Feb 2012 | 7:08 pm CET

New Paper in Press HMGB1 Impairs P. aeruginosa Clearance in CF Lungs Quelle: Molecular Medicine | 6 Feb 2012 | 3:47 pm CET

New Paper in Press Microglia Stimulation of Glioblastoma Invasion Quelle: Molecular Medicine | 6 Feb 2012 | 3:45 pm CET

Drosophila as a Model to Study the Genetic Mechanisms of Obesity-Associated Heart Dysfunction. Abstract Obesity and cardiovascular disease are among the world's leading causes of death, especially in Western countries where consumption of high caloric food is commonly accompanied by low physical activity. This lifestyle often leads to energy imbalance, obesity, diabetes, and their associated metabolic disorders, including cardiovascular diseases. It has become increasingly recognized that obesity and cardiovascular disease are metabolically linked, and a better understanding of this relationship requires that we uncover the fundamental genetic mechanisms controlling obesity-related heart dysfunction, a goal that has been difficult to achieve in higher organisms with intricate metabolic complexity. However, the high degree of evolutionary conservation of genes and signaling pathways allows researchers to use lower animal models such as Drosophila, which is the simplest genetic model with a heart, to uncover the mechanistic basis of obesity-related heart disease and its likely relevance to humans. Here we discuss recent advances made by using the power of the Drosophila as a powerful model to investigate the genetic pathways by which a high fat diet may lead to heart dysfunction. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 4 Feb 2012 | 3:52 pm CET

The E3 ubiquitin ligase TRIM11 mediates the degradation of congenital central hypoventilation syndrome-associated polyalanine-expanded PHOX2B Abstract   Expansions of a polyalanine (polyA) stretch in the coding region of the PHOX2B gene cause congenital central hypoventilation syndrome (CCHS), a neurocristopathy characterized by the absence of adequate control of autonomic breathing. Expansion of polyA in PHOX2B leads to protein misfolding and accumulation into inclusions. The mechanisms that regulate mutant protein degradation and turnover have been poorly elucidated. Here, we investigate the regulation of degradation of wild-type and polyA-expanded PHOX2B. We show that expanded PHOX2B is targeted for degradation through the ubiquitin–proteasome system, resulting in lowered levels of the mutant protein relative to its wild-type counterpart. Moreover, we show that mutant PHOX2B forms ubiquitin-positive inclusions, which sequester wild-type PHOX2B. This sequestration correlates with reduced transcriptional activity of endogenous wild-type protein in neuroblastoma cells. Finally, we show that the E3 ubiquitin ligase TRIM11 plays a critical role in the clearance of mutant PHOX2B through the proteasome. Importantly, clearance of mutant PHOX2B by TRIM11 correlates with a rescue of PHOX2B transcriptional activity. We propose that CCHS is partially caused by a dominant-negative effect of expanded PHOX2B due to the retention of the wild-type protein in pathogenic aggregates. Our results demonstrate that TRIM11 is a novel modifier of mutant PHOX2B toxicity and represents a potential therapeutic target for CCHS. Content Type Journal Article Category Original Article Pages 1-11 DOI 10.1007/s00109-012-0868-1 Authors Sara Parodi, Laboratory of Molecular Genetics, G. Gaslini Institute, 16148 Genoa, Italy Eleonora Di Zanni, Laboratory of Molecular Genetics, G. Gaslini Institute, 16148 Genoa, Italy Simona Di Lascio, Department of Pharmacology, School of Medicine, University of Milan and CNR—Institute of Neuroscience, Milan, Italy Paola Bocca, Laboratory of Oncology, G. Gaslini Institute, Genoa, Italy Ignazia Prigione, Laboratory of Oncology, G. Gaslini Institute, Genoa, Italy Diego Fornasari, Department of Pharmacology, School of Medicine, University of Milan and CNR—Institute of Neuroscience, Milan, Italy Maria Pennuto, Department of Neuroscience and Brain Technologies, Italian Institute of Technology, Genoa, Italy Tiziana Bachetti, Laboratory of Molecular Genetics, G. Gaslini Institute, 16148 Genoa, Italy Isabella Ceccherini, Laboratory of Molecular Genetics, G. Gaslini Institute, 16148 Genoa, Italy Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 3 Feb 2012 | 6:52 pm CET

microRNAs in the regulation of dendritic cell functions in inflammation and atherosclerosis Abstract   Atherosclerosis has been established as a chronic inflammatory disease of the vessel wall. Among the mononuclear cell types recruited to the lesions, specialized dendritic cells (DCs) have gained increasing attention, and their secretory products and interactions shape the progression of atherosclerotic plaques. The regulation of DC functions by microRNAs (miRNAs) may thus be of primary importance in disease. We here systematically summarize the biogenesis and functions of miRNAs and provide an overview of miRNAs in DCs, their targets, and potential implications for atherosclerosis, with a particular focus on the best characterized miRNAs in DCs, namely, miR-155 and miR-146. MiRNA functions in DCs range from regulation of lipid uptake to cytokine production and T cell responses with a complex picture emerging, in which miRNAs cooperate or antagonize DC behavior, thereby promoting or counterbalancing inflammatory responses. As miRNAs regulate key functions of DCs known to control atherosclerotic vascular disease, their potential as a therapeutic target holds promise and should be attended to in future research. Content Type Journal Article Category Review Pages 1-9 DOI 10.1007/s00109-012-0864-5 Authors Martin Busch, Rudolf-Virchow-Center/DFG Research Center for Experimental Biomedicine, University of Würzburg, Josef-Schneider Str. 2, Haus D15, 97080 Würzburg, Germany Alma Zernecke, Rudolf-Virchow-Center/DFG Research Center for Experimental Biomedicine, University of Würzburg, Josef-Schneider Str. 2, Haus D15, 97080 Würzburg, Germany Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 3 Feb 2012 | 6:52 pm CET

Structural insights into a human anti-IFN antibody exerting therapeutic potential for systemic lupus erythematosus Abstract   Increasing evidences suggest that the type I interferon α (IFNα) plays a critical role in the etiopathogenesis of systemic lupus erythematosus (SLE), which makes it a promising therapeutic target for the treatment of the disease. By screening a large size non-immune human antibody library, we have developed a human single-chain antibody (ScFv) AIFNα1bScFv01 and corresponding whole antibody AIFNα1bIgG01 to human interferon α1b (IFNα1b) with high specificity and high affinity. The IgG antibody could down-regulate the expression of ISG15 and IFIT-1 induced by either recombinant IFNα1b or naïve IFNα from SLE patients’ sera, and reduced total serum IgG and IgM antibodies level in a pristane-primed lupus-like mouse model. The crystal structure of AIFNα1bScFv01-IFNα1b complex solved to 2.8 Å resolution revealed that both Pro26-Gln40 region in loop AB and Glu147-Arg150 region in helix E of IFNα1b contribute to binding with AIFNα1bScFv01. Four residues of above two regions (Leu30, Asp32, Asp35 and Arg150) are critical for the formation of antigen–antibody complexes. AIFNα1bScFv01 shares partial epitopes of IFNα1b with its receptor IFNAR2 but with much higher binding affinity to IFNα1b than IFNAR2. Thus, AIFNα1bIgG01 exhibits its neutralizing activity through competition with IFNAR2 to bind with IFNα and prevents the activation of IFNα-mediated signaling pathway. Our results highlight the potential use of the human antibody for modulating the activity of IFNα in SLE. Content Type Journal Article Category Original Article Pages 1-10 DOI 10.1007/s00109-012-0866-3 Authors Songying Ouyang, National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101 China Bin Gong, National Institute for Viral Diseases Control and Prevention, China CDC, Beijing, 102206 China Jin-Zhi Li, Shanghai Institute of Immunology, Institute of Medical Science, Shanghai Jiaotong University School of Medicine, Shanghai, 200025 China Li-Xia Zhao, National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101 China Wei Wu, National Institute for Viral Diseases Control and Prevention, China CDC, Beijing, 102206 China Fu-Shun Zhang, National Institute for Viral Diseases Control and Prevention, China CDC, Beijing, 102206 China Lina Sun, National Institute for Viral Diseases Control and Prevention, China CDC, Beijing, 102206 China Shu-Jun Wang, Shanghai Institute of Immunology, Institute of Medical Science, Shanghai Jiaotong University School of Medicine, Shanghai, 200025 China Meng Pan, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200025 China Chuan Li, National Institute for Viral Diseases Control and Prevention, China CDC, Beijing, 102206 China Wenguang Liang, National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101 China Neil Shaw, National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101 China Jie Zheng, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200025 China Guo-Ping Zhao, Shanghai-MOST Key Laboratory for Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, 201203 China Ying Wang, Shanghai Institute of Immunology, Institute of Medical Science, Shanghai Jiaotong University School of Medicine, Shanghai, 200025 China Zhi-Jie Liu, National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101 China Mifang Liang, National Institute for Viral Diseases Control and Prevention, China CDC, Beijing, 102206 China Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 3 Feb 2012 | 6:52 pm CET

Overexpression of mouse TTF-2 gene causes cleft palate Abstract In humans, mutations of the gene encoding for thyroid transcription factor-2 (TTF-2 or FOXE1) result in Bamforth syndrome. Bamforth syndrome is characterized by agenesis, cleft palate, spiky hair, and choanal atresia. TTF-2 null mice (TTF-2-/-) also exhibit cleft palate, suggesting its involvement in the palatogenesis. However, the molecular pathology and genetic regulation by TTF2 are largely unknown. In the present study, the recombinant expression vector pBROAD3-TTF-2 containing the promoter of the mouse ROSA26 gene was created to form the structural gene of mouse TTF-2 and was microinjected into the male pronuclei of fertilized ova. Sequence analysis confirmed that the TTF-2 transgenic mouse model was established successfully. The transgenic mice displayed a phenotype of cleft palate. In addition, we found that TTF-2 was highly expressed in the medial edge epithelium (MEE) from the embryonic day 12.5 (E12.5) to E14.5 in TTF-2 transgenic mice. These observations suggest that overexpression of TTF-2 during palatogenesis may contribute to formation of cleft palate. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 3 Feb 2012 | 6:41 pm CET

Osteoinduction and survival of osteoblasts and bone-marrow stromal cells in 3D biphasic calcium phosphate scaffolds under static and dynamic culture conditions Abstract In many tissue engineering approaches, the basic difference between in vitro and in vivo conditions for cells within three-dimensional (3D) constructs is the nutrition flow dynamics. To achieve comparable results in vitro, bioreactors are advised for improved cell survival, as they are able to provide a controlled flow through the scaffold. We hypothesize that a bioreactor would enhance long term differentiation conditions of osteogenic cells in 3D scaffolds. To achieve this either primary rat osteoblasts or bone marrow stromal cells (BMSC) were implanted on uniform-sized biphasic calcium phosphate (BCP) scaffolds produced by a 3D printing method. Three types of culture conditions were applied: static culture without osteoinduction (Group A); static culture with osteoinduction (Group B); dynamic culture with osteoinduction (Group C). After three and six weeks, the scaffolds were analyzed by ALP, dsDNA amount, SEM, fluorescent labeled live-dead assay, and real-time RT-PCR in addition to weekly alamarBlue assays. With osteoinduction, increased ALP values and calcium deposition are observed; however, under static conditions, a significant decrease in the cell number on the biomaterial is observed. Interestingly, the bioreactor system not only reversed the decreased cell numbers, but also increased their differentiation potential. We conclude from this study that a continuous flow bioreactor not only preserves the number of osteogenic cells, but also keeps their differentiation ability in balance providing a suitable cell-seeded scaffold product for applications in regenerative medicine. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 3 Feb 2012 | 6:37 pm CET

Potential biomarkers in psychiatry: focus on the cholesterol system Abstract Measuring biomarkers to identify and assess illness is a strategy growing in popularity and relevance. Although already in clinical use for treating and predicting cancer, no biological measurement is used clinically for any psychiatric disorder. Biomarkers could predict the course of a medical problem, and aid in determining how and when to treat. Several studies have indicated that of candidate psychiatric biomarkers detected using proteomic techniques, cholesterol and associated proteins, specifically apolipoproteins (Apos), may be of interest. Cholesterol is necessary for brain development and its synthesis continues at a lower rate in the adult brain. Apos are the protein component of lipoproteins responsible for lipid transport. There is extensive evidence that the levels of cholesterol and Apos may be disturbed in psychiatric disorders, including autistic spectrum disorders (ASD). Here we describe putative serum biomarkers for psychiatric disorders, and the role of cholesterol and Apos in central nervous system (CNS) disorders. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 3 Feb 2012 | 6:37 pm CET

A logistic model for the detection of circulating tumor cells in human metastatic colorectal cancer Abstract The accuracy in the diagnosis of metastatic colorectal cancer (mCRC) represents one of the challenges in the clinical management of patients. The detection of Circulating Tumor Cells (CTC) is becoming a promising alternative to current detection techniques as it focuses on one of the players of the metastatic disease and it should provide with more specific and sensitive detection rates. Here, we describe an improved method of detection of CTC from mCRC patients by combining immune-enrichment, optimal purification of RNA from very low cell numbers, and the selection of accurate PCR probes. As a result, we obtained a logistic model that combines GAPDH and VIL1 normalized to CD45 rendering powerful results in the detection of CTC from mCRC patients (AUROC value 0.8599). We further demonstrated the utility of this model at the clinical setting, as a reliable prognosis tool to determine progression free survival in mCRC patients. Overall, we developed a strategy that ameliorates the specificity and sensitivity in the detection of CTC, resulting in a robust and promising logistic model for the clinical management of metastatic colorectal cancer patients. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 3 Feb 2012 | 6:36 pm CET

Genistein as a potential inducer of the anti-atherogenic enzyme paraoxonase-1: studies in cultured hepatocytes in vitro and in rat liver in vivo Abstract A number of cardioprotective effects, including the reduction of the LDL particle to oxidation, have been attributed to dietary soy isoflavones. Paraoxonase 1 (PON1), an enzyme mainly synthesized in the liver, may exhibit anti-atherogenic activity by protecting LDL from oxidation. Thus, dietary and pharmacological inducers of PON1 may reduce cardiovascular disease risk. Using a luciferase reporter gene assay we screened different flavonoids for their ability to induce PON1 in Huh7 hepatocytes in culture. Genistein was the most potent flavonoid regarding its PON1 inducing activity, followed by daidzein, luteolin, isorhamnetin and quercetin. Other flavonoids such as naringenin, cyanidin, malvidin and catechin showed only little or no PON1 inducing activity. Genistein-mediated PON1 transactivation was partly inhibited by the estrogen-receptor antagonist fulvestrant as well as by the aryl hydrocarbon receptor antagonist 7-ketocholesterol. In contrast to genistein, the conjugated genistein metabolites genistein-7-O-β-D-glucuronide, genistein-7-O-sulfate and genistein-7,4’-O-disulfate were only a weak inducers of PON1 transactivation. Accordingly, dietary genistein supplementation (2 g/kg diet over 3 weeks) in growing rats did not increase hepatic PON1 mRNA and protein levels as well as plasma PON1 activity. Thus, genistein may be a PON1 inducer in cultured hepatocytes in vitro but not in rats in vivo. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 3 Feb 2012 | 6:36 pm CET

AL3810, a multi-tyrosine kinase inhibitor, exhibits potent anti-angiogenic and antitumor activity via targeting VEGFR, FGFR, and PDGFR Abstract Angiogenesis plays an important role in neoplastic transformation and progression as well as in the metastasis process of most human cancers. Herein, we identified AL3810 as a novel and orally bioavaliable small molecular inhibitor with potent inhibitory activity against multiple tyrosine kinases involved in the process of angiogenesis. We found that AL3810 substantially inhibited the autophosphorylation of VEGFR2, PDGFRβ and FGFR1 in endothelial cells. Moreover, AL3810 exhibited potent anti-angiogenesis activity, manifested by significant inhibition of microvessel outgrowth of rat arterial ring and chickallantochorion membrane (CAM) in ex vivo angiogenesis models. Daily dosing of AL3810 has shown broad-spectrum anti-tumor activity in human kidney, pancreas, liver cancer xenograft models. Importantly, immunohistochemistry results demonstrated that the antitumor activity of AL3810 was closely correlated with its anti-angiogenesis activity, as evidenced by a decreased microvessel area and reduced microvessel numbers in tumor tissues. The overall pharmacological profiles of AL3810 are superior to sorafenib. The clinical trials of AL3810 will soon be launched in China. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 3 Feb 2012 | 6:35 pm CET

Vestibular Regeneration – Experimental Models and Clinical Implications Abstract Therapies aimed at the protection and/or regeneration of inner ear hair cells are of great interest given the significant monetary and quality of life impact of balance disorders. Different viral vectors have been shown to transfect various cell types in the inner ear. The past decade has provided tremendous advances in the use of adenoviral vectors to achieve targeted treatment delivery. Several routes of delivery have been identified in order to introduce vectors into the inner ear while minimizing injury to surrounding structures. Recently, the transcription factor Atoh1 was determined to play a critical role in hair cell differentiation. Adenoviral-mediated overexpression of Atoh1 in culture and in vivo has demonstrated the ability to regenerate vestibular hair cells by causing transdifferentiation of neighboring epithelial-supporting cells. Functional recovery of the vestibular system has also been documented following adenoviral-induced Atoh1 overexpression. Experiments demonstrating gene transfer in human vestibular epithelial cells reveal that the human inner ear is a suitable target for gene therapy. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 3 Feb 2012 | 6:35 pm CET

Top 10 Downloads for January Quelle: Molecular Medicine | 2 Feb 2012 | 5:23 pm CET

New Paper in Press HMGB1 Impairs P. aeruginosa Clearance in CF Lungs Quelle: Molecular Medicine | 2 Feb 2012 | 3:52 pm CET

High-molecular weight Aβ oligomers and protofibrils are the predominant Aβ species in the native soluble protein fraction of the AD brain Abstract Alzheimer’s disease (AD) is characterized by the aggregation and deposition of amyloid β protein (Aβ) in the brain. Soluble Aβ oligomers are thought to be toxic. To investigate the predominant species of Aβ protein that may play a role in AD pathogenesis, we performed biochemical analysis of AD and control brains. Sucrose buffer-soluble brain lysates were characterized in native form using blue native (BN)-PAGE and also in denatured form using SDS-PAGE followed by Western blot analysis. BN-PAGE analysis revealed a high-molecular weight smear (>1000 kD) of Aβ42-positive material in the AD brain, whereas low-molecular weight and monomeric Aβ species were not detected. SDS-PAGE analysis, on the other hand, allowed the detection of prominent Aβ monomer and dimer bands in AD cases but not in controls. Immunoelectron microscopy of immunoprecipitated oligomers and protofibrils/fibrils showed spherical and protofibrillar Aβ-positive material, thereby confirming the presence of high-molecular weight Aβ (hiMWAβ) aggregates in the AD brain. In vitro analysis of synthetic Aβ40- and Aβ42 preparations revealed Aβ fibrils, protofibrils, and hiMWAβ oligomers that were detectable at the electron microscopic level and after BN-PAGE. Further, BN-PAGE analysis exhibited a monomer band and less prominent low-molecular weight Aβ (loMWAβ) oligomers. In contrast, SDS-PAGE showed large amounts of loMWAβ but no hiMWAβ40 and strikingly reduced levels of hiMWAβ42. These results indicate that hiMWAβ aggregates, particularly Aβ42 species, are most prevalent in the soluble fraction of the AD brain. Thus, soluble hiMWAβ aggregates may play an important role in the pathogenesis of AD either independently or as a reservoir for release of loMWAβ oligomers. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Methamphetamine alters occludin expression via NADPH oxidase-induced oxidative insult and intact caveolae Abstract Methamphetamine (METH) is a drug of abuse with neurotoxic and vascular effects that may be mediated by reactive oxygen species (ROS). However, potential sources of METH-induced generation of ROS are not fully understood. This study is focused on the role of NAD(P)H oxidase (NOX) in METH-induced dysfunction of brain endothelial cells. Treatment with METH induced a time-dependent increase in phosphorylation of NOX subunit p47, followed by its binding with gp91 and p22, and the formation of an active NOX complex. An increase in NOX activity was associated with elevated production of ROS, alterations of occludin levels and increased transendothelial migration of monocytes. Inhibition of NOX by NSC 23766 attenuated METH-induced ROS generation, changes in occludin protein levels and monocyte migration. Because an active NOX complex is localized to caveolae, we next evaluated the role of caveolae in METH-mediated toxicity to brain endothelial cells. Treatment with METH induced phosphorylation of ERK1/2 and caveolin-1 protein. Inhibition of ERK1/2 activity or caveolin-1 silencing protected against METH-induced alterations of occludin levels. These findings indicate an important role of NOX and functional caveolae in METH-induced oxidative stress in brain endothelial cells that contribute to the subsequent alterations of occludin levels and transendothelial migration of inflammatory cells. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

ENDOGLIN/CD105 is expressed in KIT positive cells in the gut and in gastrointestinal stromal tumours Abstract ENDOGLIN/CD105 (ENG) is a transmembrane glycoprotein and an auxiliary unit of the transforming growth factor-β (TGF-β); receptor, expressed predominantly in vascular endothelium. Noteworthy, Eng mRNA expression has been reported also in Kit+ interstitial cells of Cajal (ICC) in the mouse intestine. Gastrointestinal stromal tumours (GIST) are thought to derive from ICC. Here we have investigated Eng expression in the KitK641E mouse GIST model, in human GIST and in the Ba/F3 cell model. In wild type (WT) mouse antrum, Eng immunoreactivity (-ir) was detected in CD34+/CD31+ endothelium and in Kit+ ICC. In KitK641E mice, hyperplasia of Kit+ cells made Eng-ir even more evident. Quantitative PCR confirmed the increased expression of Eng transcript in KitK641E mice. On human GIST TMA, 26/49 cases stained positive for ENG. Strong ENG staining was associated with malignant and high-risk tumours. ENG negative cases were predominantly of the epithelioid type or harboured PDGFRA mutation. In vitro, Eng mRNA was up-regulated in Ba/F3 cell lines stably expressing various oncogenic Kit mutations (K641E, del559, del814). This effect appeared to be independent of Kit activation, as neither the stimulation of WT Kit by its ligand SCF, nor the inhibition of Kit autophosphorylation by imatinib mesylate in oncogenic mutants, altered Eng expression. Elevated Eng expression in Kit oncogenic mutants appeared rather to be indirectly mediated by DNA hypomethylation, because treatment with the demethylating agent 5-Aza/dC increased Eng mRNA expression in KitWT cells. ENG expression in ICC and in GIST deserves further consideration as ENG is emerging as a potential target for cancer therapy. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Progesterone enhances vascular endothelial cell migration via activation of focal adhesion kinase Abstract The mechanisms of progesterone on endothelial cell motility are poorly investigated. Previously we showed that progesterone stimulated endothelial cell migration via the activation of actin-binding protein moesin, leading to actin cytoskeleton remodelling and the formation of cell membrane structures required for cell movement. In this study, we investigated the effects of progesterone on the formation of focal adhesion complexes, which provide anchoring sites for cell movement. In cultured human umbilical endothelial cells, progesterone enhanced focal adhesion kinase (FAK) phosphorylation at Tyr397 in a dose- and time-dependent manner. Several signalling inhibitors interfered with progesterone-induced FAK activation, including progesterone receptor (PR) antagonist ORG 31710, specific c-Src kinase inhibitor PP2, phosphatidylinosital-3 kinase (PI3K) inhibitor wortmannin as well as ρ-associated kinase (ROCK-2) inhibitor Y27632. It suggested that PR, c-Src, PI3K and ROCK-2 are implicated in this action. In line with this, we found that progesterone rapidly promoted c-Src/PI3K/Akt activity, which activated the small GTPase RhoA/ρ-associated kinase (ROCK-2) complex, resulting in FAK phosphorylation. In the presence of progesterone, endothelial cells displayed enhanced horizontal migration, which was reversed by small interfering RNAs abrogating FAK expression. In conclusion, progesterone promotes endothelial cell movement via the rapid regulation of FAK. These findings provide new information on the biological actions of progesterone on human endothelial cells that are relevant for vascular function. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Calcineurin regulation of cytoskeleton organization: a new paradigm to analyse the effects of calcineurin inhibitors on the kidney Abstract •  Introduction •  Calcineurin effects on neuronal cytoskeleton •  Calcineurin effects on myocytes •  Intracellular pathway involved in calcineurin-induced effects on cytoskeletal organization -  Calcineurin/NFAT pathway -  Calcineurin/NFAT in neuronal elongation -  Calcineurin/NFAT in cardiac hypertrophy -  Calcineurin/actin-associated proteins pathway •  Calcineurin effects on kidney cells    cytoskeleton •  Conclusion Calcineurin is a serine/threonine phosphatase originally involved in the immune response but is also known for its role as a central mediator in various non-immunological intracellular signals. The nuclear factor of activated T cell (NFAT) proteins are the most widely described substrates of calcineurin, but ongoing work has uncovered other substrates among which are the cytoskeleton organizing proteins (i.e. cofilin, synaptopodin, WAVE-1). Control over cytoskeletal proteins is of outmost interest because the phenotypic properties of cells are dependent on cytoskeleton architecture integrity, while rearrangements of the cytoskeleton are implicated in both physiological and pathological processes. Previous works investigating the role of calcineurin on the cytoskeleton have focused on neurite elongation, myocyte hypertrophic response and recently in kidney cells structure. Nuclear factor of activated T cell activation is expectedly identified in the signalling pathways for calcineurin-induced cytoskeleton organization, however new NFAT-independent pathways have also been uncovered. The aim of this review is to summarize the current knowledge on the effects of calcineurin on cytoskeletal proteins and related intracellular pathways. These newly described properties of calcineurin on cytoskeletal proteins may explain some of the beneficial or deleterious effects observed in kidney cells associated with the use of the calcineurin inhibitors, cyclosporine and tacrolimus. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

A limited role for regulatory T cells in post-ischemic neovascularization Abstract Recently, it was demonstrated that arteriogenesis is enhanced in mice deficient in regulatory T cells (CD4+CD25+FoxP3+ T cell), which can suppress effector T cell responses. The present study investigates the effects of these regulatory T cells on arteriogenesis in more detail by either specific expanding or depleting regulatory T cells. Hind limb ischemia was induced by electro-coagulation of the femoral artery in mice. Regulatory T cells were either expanded by injecting mice with a complex of interleukin (IL)-2 with the IL-2 monoclonal antibody JES6–1, or depleted by anti-CD25 antibody or diphtheria toxin injections in DEREG mice (depletion of regulatory T cells). Blood flow restoration was monitored using laser Doppler perfusion imaging. Collateral arteries were visualized by immunohistochemistry. Regulatory T cell expansion led to a moderate though significant suppression of blood flow restoration after ischemia induction. Surprisingly, depletion of regulatory T cells resulted in minor increase on blood flow recovery. However, collateral and capillary densities in the post-ischemic skeletal muscle were significantly increased in DEREG mice depleted for regulatory T cells. The presence of regulatory T cells after ischemia induction when analysed in non-depleted DEREG mice could be demonstrated by green fluorescent protein staining only in lymph nodes in the ischemic area, and not in the ischemic muscle tissue. The current study demonstrates that, even under conditions of major changes in regulatory T cell content, the contribution of regulatory T cells to the regulation of the arteriogenic response is only moderate. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Tracheal telocytes Abstract Recently, a novel type of stromal cell – the telocytes (TC) – was identified in mouse trachea. These cells are known to possess the ultrastructural characteristics, which support their role in intercellular signaling. We found TC in all stromal compartments of the tracheal wall. TC with long prolongations (telopodes, Tp) were lining longitudinally the collagen bundles, and were serially arranged (end-to-end connections of Tp were found). Noteworthy, Tp frequently establish stromal synapses with mast cells (MC). Primary cilia were also identified in TC. In conclusion, tracheal TC could be involved in the tracheal regulation (e.g. secretion, contractility). The tandem TC-MC deserves further investigations. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Elevated levels of hypoxia-inducible microRNA-210 in pre-eclampsia: new insights into molecular mechanisms for the disease Abstract Pre-eclampsia is a leading cause of maternal and foetal morbidity and mortality worldwide. Insufficient uteroplacental oxygenation is believed to be responsible for the disease. However, what molecular events involve in hypoxic responses and how they affect placental development remain unclear. Recently, miRNAs have emerged as a new class of molecules in response to hypoxia. We show here that the expression of microRNA-210 (mir-210) is up-regulated in patients with pre-eclampsia, as well as in trophoblast cells cultured under hypoxic conditions. Ectopic expression of mir-210 inhibited the migration and invasion capability of trophoblast cells. Ephrin-A3 and Homeobox-A9, which related with cell migration and vascular remodelling, were then experimentally validated as the functional targets of mir-210 both in vivo and in vitro. Using luciferase reporter, chromatin immunoprecipitation (ChIP) and small interfering RNA (siRNA) experiments, we finally identified a new transcriptional mechanism that the overexpression of mir-210 under hypoxia was regulated by NF-κB transcriptional factor p50, apart from the well-known HIF 1α. Taken together, our study implicates an important role for mir-210 in the molecular mechanism of pre-eclampsia. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Apoptosis related protein-1 triggers melanoma cell death via interaction with the juxtamembrane region of p75 neurotrophin receptor Abstract Although chemotherapeutic drugs could theoretically target all metastatic sites, current treatments do not provide complementary therapeutics. Therefore, the development of an alternative approach replacing the traditional therapy is urgently needed. To assess the killing efficiency of the functionally identified apoptosis-related protein (APR)-1 in melanoma cells, we established a system for the regulated expression of APR-1. The induction of APR-1 expression caused apoptosis of melanoma cells via the interaction with the juxtamembrane region of p75 neurotrophin receptor (p75NTR), and possible also via the competition with tumour necrosis factor receptor-associated factor-6 (TRAF6) and the catalytic receptor of neurotrophin (Trk) for the same p75NTR interacting site. The accumulation of APR-1 in melanoma cells may block the physical association of p75NRT with TRAF6 and/or Trk, leading to the disruption of both NF-κB and extracellular signal-regulated kinase (ERK) pathways. Also, accumulation of APR-1 protein enhanced the activity of both c-Jun-N-terminal kinase (JNK) and p38 pathways. However, the analysis of APR-1-modulated pathways demonstrated the involvement of apoptosis-regulating kinase 1-JNK/p38 pathway in the induction of Bax expression leading to both mitochondrial dysregulation [as demonstrated by the loss of mitochondrial membrane potential, the release of both cytochrome c and apoptosis-inducing factor into cytoplasm, and cleavage of caspase-9, caspase-3 and poly (ADP-ribose) polymerase (PARP)] and endoplasmic reticulum stress as demonstrated by the increase of intracellular Ca2+ release. Thus, besides the analysis of its pro-apoptotic function, our data provide insight into the molecular mechanism of APR-1-induced apoptosis of melanoma cells. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Issue Information Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Immunohistochemical analysis of myenteric ganglia and interstitial cells of Cajal in ulcerative colitis Abstract Ulcerative colitis (UC) is an inflammatory bowel disease with alterations of colonic motility, which influence clinical symptoms. Although morpho-functional abnormalities in the enteric nervous system have been suggested, in UC patients scarce attention has been paid to possible changes in the cells that control colonic motility, including myenteric neurons, glial cells and interstitial cells of Cajal (ICC). This study evaluated the neural-glial components of myenteric ganglia and ICC in the colonic neuromuscular compartment of UC patients by quantitative immunohistochemical analysis. Full-thickness archival samples of the left colon were collected from 10 patients with UC (5 males, 5 females; age range 45–62 years) who underwent elective bowel resection. The colonic neuromuscular compartment was evaluated immunohistochemically in paraffin cross-sections. The distribution and number of neurons, glial cells and ICC were assessed by anti-HuC/D, -S100β and -c-Kit antibodies, respectively. Data were compared with findings on archival samples of normal left colon from 10 sex- and age-matched control patients, who underwent surgery for uncomplicated colon cancer. Compared to controls, patients with UC showed: (i) reduced density of myenteric HuC/D+ neurons and S100β+ glial cells, with a loss over 61% and 38%, respectively, and increased glial cell/neuron ratio; (ii) ICC decrease in the whole neuromuscular compartment. The quantitative variations of myenteric neuro-glial cells and ICC indicate considerable alterations of the colonic neuromuscular compartment in the setting of mucosal inflammation associated with UC, and provide a morphological basis for better understanding the motor abnormalities often observed in UC patients. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

STAT1 deficiency in the heart protects against myocardial infarction by enhancing autophagy Abstract Previous studies have shown that the transcription factor signal transducer and activator of transcription 1 (STAT1) activation is increased in primary cardiac myocytes exposed to simulated ischaemia/reperfusion injury. This promotes apoptotic cell death by enhancing the expression of pro-apoptotic proteins. Autophagy has been demonstrated to play a cardioprotective role in the heart following myocardial infarction (MI). We therefore investigated the role of STAT1 in the intact heart subjected to MI and examined the contribution of autophagy in modulating the protective effect of STAT1 after MI injury. STAT1-deficient hearts had significantly smaller infarcts than wild-type hearts and this correlated with increased levels of autophagy shown by light chain 3 (LC3)-I/LC3-II conversion, and up-regulation of Atg12 and Beclin 1. Moreover, pre-treatment with the autophagy inhibitor 3-methyladenine reversed the cardioprotection observed in the STAT1-deficient hearts. These results reveal a new function of STAT1 in the control of autophagy and indicate a cross-talk between the cardioprotective versus the damaging effects of STAT1 in the intact heart exposed to MI injury. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients Abstract Early detection of resistance to platinum-based therapy is critical for improving the treatment of ovarian cancers. We have previously found that increased expression of annexin A3 is a mechanism for platinum resistance in ovarian cancer cells. Here we demonstrate that annexin A3 can be detected in the culture medium of ovarian cancer cells, particularly these cells that express high levels of annexin A3. Levels of annexin A3 were then determined in sera from ovarian cancer patients using an enzyme-linked immunosorbent assay. Compared with those from normal donors, sera from ovarian cancer patients contain significantly higher levels of annexin A3. Furthermore, serum levels of annexin A3 were significantly higher in platinum-resistant patients than in platinum-sensitive patients. To gain insight into the mechanism of secretion, the ovarian cancer cell lines were examined using both transmission electron microscopy and immunoelectron microscopy. Compared with parent cells, there are significantly more vesicles in the cytoplasm of ovarian cancer cells that express high levels of annexin A3, and at least some vesicles are annexin A3-positive. Moreover, some vesicles appear to be fused with the cell membrane, suggesting that annexin A3 secretion may be associated with exocytosis and the release of exosomes. This is supported by our observation that ovarian cancer cells expressing higher levels of annexin A3 released increased numbers of exosomes. Furthermore, annexin A3 can be detected in exosomes released from cisplatin-resistant cells (SKOV3/Cis) by immunoblotting and immunoelectron microscopy. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Atorvastatin activates heme oxygenase-1 at the stress response elements Abstract Statins are known to inhibit growth of a number of cancer cells, but their mechanism of action is not well established. In this study, human prostate adenocarcinoma PC-3 and breast adenocarcinoma MCF-7 cell lines were used as models to investigate the mechanism of action of atorvastatin, one of the statins. Atorvastatin was found to induce apoptosis in PC-3 cells at a concentration of 1 μM, and in MCF-7 cells at 50 μM. Initial survey of possible pathway using various pathway-specific luciferase reporter assays showed that atorvastatin-activated antioxidant response element (ARE), suggesting oxidative stress pathway may play a role in atorvastatin-induced apoptosis in both cell lines. Among the antioxidant response genes, heme oxygenase-1 (HO-1) was significantly up-regulated by atorvastatin. Pre-incubation of the cells with geranylgeranyl pyrophosphate blocked atorvastatin-induced apoptosis, but not up-regulation of HO-1, suggesting that atorvastatin-induced apoptosis is dependent on GTPase activity and up-regulation of HO-1 gene is not. Six ARE-like elements (designated StRE1 [stress response element] through StRE6) are present in the HO-1 promoter. Atorvastatin was able to activate all of the elements. Because these StRE sites are present in clusters in HO-1 promoter, up-regulation of HO-1 by atorvastatin may involve multiple StRE sites. The role of HO-1 in atorvastatin-induced apoptosis in PC-3 and MCF-7 remains to be studied. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Alterations in the contractile phenotype of the bladder: lessons for understanding physiological and pathological remodelling of smooth muscle Abstract •  Introduction •  Alterations in bladder contractility with normal development •  Assessment of bladder function in genetically modified mice •  A whole organ perspective on the bladder’s response to pBOO -  Clinical observations •  Experimental observations •  In vivo measures of pBOO in a rabbit model •  Relating muscle strip studies to whole organ function •  Myosin light chain phosphorylation •  Regulation of myosin light chain de-phosphorylation •  Alterations in the thick contractile filaments -  SM1 and SM2 isoforms -  SM-B and SM-A isoforms •  Regulatory light chain •  Alterations in the thin contractile filaments •  Alterations in tension transfer complex The contractile properties of the urinary bladder are changed by the conditions of normal development and partial bladder outlet obstruction. This change in the contractile phenotype is accompanied by changes in the regulatory cascades and filaments that regulate contractility. This review focuses on such changes during the course of normal development and in response to obstruction. Our goal is to discuss the experimental evidence that has accumulated from work in animal models and correlate these findings with the human voiding phenotype. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

α-Lipoic acid attenuates vascular calcification via reversal of mitochondrial function and restoration of Gas6/Axl/Akt survival pathway Abstract Vascular calcification is prevalent in patients with chronic kidney disease and leads to increased cardiovascular morbidity and mortality. Although several reports have implicated mitochondrial dysfunction in cardiovascular disease and chronic kidney disease, little is known about the potential role of mitochondrial dysfunction in the process of vascular calcification. This study investigated the effect of α-lipoic acid (ALA), a naturally occurring antioxidant that improves mitochondrial function, on vascular calcification in vitro and in vivo. Calcifying vascular smooth muscle cells (VSMCs) treated with inorganic phosphate (Pi) exhibited mitochondrial dysfunction, as demonstrated by decreased mitochondrial membrane potential and ATP production, the disruption of mitochondrial structural integrity and concurrently increased production of reactive oxygen species. These Pi-induced functional and structural mitochondrial defects were accompanied by mitochondria-dependent apoptotic events, including release of cytochrome c from the mitochondria into the cytosol, subsequent activation of caspase-9 and -3, and chromosomal DNA fragmentation. Intriguingly, ALA blocked the Pi-induced VSMC apoptosis and calcification by recovery of mitochondrial function and intracellular redox status. Moreover, ALA inhibited Pi-induced down-regulation of cell survival signals through the binding of growth arrest-specific gene 6 (Gas6) to its cognate receptor Axl and subsequent Akt activation, resulting in increased survival and decreased apoptosis. Finally, ALA significantly ameliorated vitamin D3-induced aortic calcification and mitochondrial damage in mice. Collectively, the findings suggest ALA attenuates vascular calcification by inhibiting VSMC apoptosis through two distinct mechanisms; preservation of mitochondrial function via its antioxidant potential and restoration of the Gas6/Axl/Akt survival pathway. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Sequential biphasic changes in claudin1 and claudin4 expression are correlated to colorectal cancer progression and liver metastasis Abstract Terminal progression of colorectal cancer (CRC) culminates in liver metastasis. To identify genes that are involved in the metastatic phenotype, cDNA microarrays were used to analyse mRNA expression profiles of colorectal carcinoma (CC)531 rat colon adenocarcinoma cells for changes related to their homing into the liver. Briefly, CC531 cells were intraportally implanted into the liver of Wag-Rij rats and re-isolated after 3, 6, 9, 14 and 21 days. Compared to control CC531 cells, claudin1 and claudin4 were among the ≥8-fold initially down-regulated genes. The co-culture of tumour cells with isolated rat hepatocytes and Kupffer cells did not induce down-regulation of either claudin1 or 4. When the environment effective on circulating tumour cells was simulated by cell culture conditions favouring their adhesion, only claudin4 showed augmented expression. Knockdown of claudin1 and claudin4 mediated by small interfering RNA caused significantly increased migration and decreased clonogenic growth of tumour cells (P < 0.05), but had no effect on their proliferation. These experimental results were paralleled by increased claudin1 and claudin4 expression in human CRC samples in Union for International Cancer Control (UICC) stages I–III, as evaluated by real-time PCR. Increased claudin4 levels were correlated with significantly reduced overall survival (log-rank test, P= 0.018). Further, significantly (P < 0.05) reduced expression of claudin1 and claudin4 was observed in stage IV and liver metastasis by immunohistochemistry. In conclusion, sequential biphasic changes in claudin1 and claudin4 expression occur during the homing of rat CC531 CRC cells to the liver. This modulation is reflected by significant changes in claudin expression in human primary and metastatic CRC. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Prometheus’s heart: what lies beneath Abstract •  Introduction •  Adult cardiomyocyte turnover •  Cardiac tissue generation: contribution of resident interstitial cells •  Signalling of miocardiogenesis: a developmental perspective •  Imaging of self-renewed myocardium •  Conclusions A heart attack kills off many cells in the heart. Parts of the heart become thin and fail to contract properly following the replacement of lost cells by scar tissue. However, the notion that the same adult cardiomyocytes beat throughout the lifespan of the organ and organism, without the need for a minimum turnover, gives way to a fascinating investigations. Since the late 1800s, scientists and cardiologists wanted to demonstrate that the cardiomyocytes cannot be generated after the perinatal period in human beings. This curiosity has been passed down in subsequent years and has motivated more and more accurate studies in an attempt to exclude the presence of renewed cardiomyocytes in the tissue bordering the ischaemic area, and then to confirm the dogma of the heart as terminally differentiated organ. Conversely, peri-lesional mitosis of cardiomyocytes were discovered initially by light microscopy and subsequently confirmed by more sophisticated technologies. Controversial evidence of mechanisms underlying myocardial regeneration has shown that adult cardiomyocytes are renewed through a slow turnover, even in the absence of damage. This turnover is ensured by the activation of rare clusters of progenitor cells interspersed among the cardiac cells functionally mature. Cardiac progenitor cells continuously interact with each other, with the cells circulating in the vessels of the coronary microcirculation and myocardial cells in auto-/paracrine manner. Much remains to be understood; however, the limited functional recovery in human beings after myocardial injury clearly demonstrates weak regenerative potential of cardiomyocytes and encourages the development of new approaches to stimulate this process. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

A therapeutic role for mesenchymal stem cells in acute lung injury independent of hypoxia-induced mitogenic factor Abstract Bone marrow mesenchymal stem cells (BM-MSCs) have therapeutic potential in acute lung injury (ALI). Hypoxia-induced mitogenic factor (HIMF) is a lung-specific growth factor that participates in a variety of lung diseases. In this study, we evaluated the therapeutic role of BM-MSC transplantation in lipopolysaccharide (LPS)- induced ALI and assessed the importance of HIMF in MSC transplantation. MSCs were isolated and identified, and untransduced MSCs, MSCs transduced with null vector or MSCs transduced with a vector encoding HIMF were transplanted into mice with LPS-induced ALI. Histopathological changes, cytokine expression and indices of lung inflammation and lung injury were assessed in the various experimental groups. Lentiviral transduction did not influence the biological features of MSCs. In addition, transplantation of BM-MSCs alone had significant therapeutic effects on LPS-induced ALI, although BM-MSCs expressing HIMF failed to improve the histopathological changes observed with lung injury. Unexpectedly, tumour necrosis factor α levels in lung tissues, lung oedema and leucocyte infiltration into lungs were even higher after the transplantation of MSCs expressing HIMF, followed by a significant increase in lung hydroxyproline content and α-smooth muscle actin expression on day 14, as compared to treatment with untransduced MSCs. BM-MSC transplantation improved LPS-induced lung injury independent of HIMF. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Tyrosine kinase gene fusions in cancer: translating mechanisms into targeted therapies Abstract •  Introduction •  Tyrosine kinase inhibitors •  Fusion gene structure and expression •  Loss of wild-type alleles •  Oligomerization triggers TK activation •  Inhibitory domain deletion •  Signalling pathways •  Recruitment of additional molecules by the fusion partner •  Stabilization and degradation •  Conclusion Tyrosine kinase fusion genes represent an important class of oncogenes associated with leukaemia and solid tumours. They are produced by translocations and other chromosomal rearrangements of a subset of tyrosine kinase genes, including ABL, PDGFRA, PDGFRB, FGFR1, SYK, RET, JAK2 and ALK. Based on recent findings, this review discusses the common mechanisms of activation of these fusion genes. Enforced oligomerization and inactivation of inhibitory domains are the two key processes that switch on the kinase domain. Activated tyrosine kinase fusions then signal via an array of transduction cascades, which are largely shared. In addition, the fusion partner provides a scaffold for the recruitment of proteins that contribute to signalling, protein stability, cellular localization and oligomerization. The expression level of the fusion protein is another critical parameter. Its transcription is controlled by the partner gene promoter, while translation may be regulated by miRNA. Several mechanisms also prevent the degradation of the oncoprotein by proteasomes and lysosomes, leading to its accumulation in cells. The selective inhibition of the tyrosine kinase activity by adenosine-5′-triphosphate competitors, such as imatinib, is a major therapeutic success. Imatinib induces remission in leukaemia patients that are positive for BCR-ABL or PDGFR fusions. Recently, crizotinib produced promising results in a subtype of lung cancers with ALK fusion. However, resistance was reported in both cases, partially due to mutations. To tackle this problem, additional levels of therapeutic interventions are suggested by the complex mechanisms of fusion tyrosine kinase activation. New approaches include allosteric inhibition and interfering with oligomerization or chaperones. Quelle: Journal of Cellular and Molecular Medicine | 1 Feb 2012 | 6:00 am CET

Myocardial regeneration by transplantation of modified endothelial progenitor cells expressing SDF-1 in a rat model Abstract Background Cell based therapy has been shown to attenuate myocardial dysfunction after myocardial infarction in different acute and chronic animal models. It has been further shown that stromal-cell derived factor-1α (SDF-1α) facilitates proliferation and migration of endogenous progenitor cells into injured tissue. Objective The aim of the presented study was to investigate the role of exogenously applied and endogenously mobilised cells in a regenerative strategy for myocardial infarction therapy. Methods and Results Lentivirally SDF-1α-infected endothelial progenitor cells (EPCs) were injected after 90 minutes of ligation and reperfusion of the left anterior descending artery (LAD) intramyocardial and intracoronary using a new rodent catheter system. 8 weeks after transplantation, echocardiography and isolated heart studies revealed a significant improvement of left ventricular function after intramyocardial application of lentiviral with SDF-1 infected EPCs compared to medium control. Intracoronary application of cells did not lead to significant differences compared to medium injected control hearts. Histology showed a significantly elevated rate of apoptotic cells and augmented proliferation after transplantation of EPCs and EPCs+SDF-1α in infarcted myocardium. Additionally, a significant increased density of CD31+ vessel structures, a lower collagen content and higher numbers of inflammatory cells after transplantation of SDF-1 transgenic cells were detectable. Conclusion intramyocardial application of lentiviral infected EPCs is associated with a significant improvement of myocardial function after infarction, in contrast to an intracoronary application. Histological results revealed a significant augmentation of neovascularization, lower collagen content, higher numbers of inflammatory cells and remarkable alterations of apoptotic/proliferative processes in infarcted areas after cell transplantation. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 30 Jan 2012 | 5:08 pm CET

The therapeutic effect of Rosuvastatin on cardiac remodeling from hypertrophy to fibrosis during the end-stage hypertension in rats Abstract Objective End-stage hypertensive heart disease is an increasing cause of cardiac mortality. Therefore, the current study focused on the cardiac remodeling from hypertrophy to fibrosis in old-aged SHRs, and explored the therapeutic effects of Rosuvastatin and its possible mechanism(s) of action. Methods SHRs at age 52 weeks were randomly divided into three groups, the first two to receive Rosuvastatin at a dose of 20mg.kg−1.d−1 and 40mg.kg−1.d−1, respectively, and the third to receive placebo, which was to be compared with Wistar-Kyoto as controls. After 2-month treatment, SBP, HW/BW% and echocardiographic features were evaluated, followed by H&E and Masson trichrome staining in conjunction with qPCR of fetal gene expressions. TUNEL assay and immunofluorescent labeling for active caspase-3 were used to detect the apoptotic cardiomyocytes. Signaling pathways involved were examined by western blot. Results Old-aged SHR developed end-stage hypertensive heart disease characterized by significant enhancement of HW/BW%, LVAWd and LVPWd, and decreased LVEF and LVFS, accompanied by cardiomyocytes enlargement and fibrosis along with activation of fetal gene program. Cardiac apoptosis increased significantly during the transition process. Rosuvastatin reduced hypertrophy significantly via AT1 Receptor-PKCβ2/α-ERK-c-fos pathway; protected myocardium against apoptosis via Akt-FOXO1, Bcl-2 family and survivin pathways, and consequently suppressed the caspase-3 activity. Conclusion The present study revealed that old-aged SHRs developed cardiac remodeling from hypertrophy to fibrosis via cardiac apoptosis during the end stage of hypertensive heart disease. These pathological changes might be the consequence of activation of AT1 Receptor-PKCβ2/α-ERK-c-fos and AKT-FOXO1/Bcl-2/survivin/Caspase3 signaling. Rosuvastatin effectively attenuated the structural changes by reversing the signaling transductions involved. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 30 Jan 2012 | 4:35 pm CET

Upregualtion of BMP-2 antagonizes TGF-β1/ROCK-enhanced cardiac fibrotic signaling through activating of Smurf1/Smad6 complex Abstract Rho-associated kinase (ROCK) plays a critical role in pressure overload-induced cardiac fibrosis, and ventricular remodeling. However, the underlying mechanism remains unclear. Here, we reported that TGF-β1-induced ROCK elevation suppressed the antagonizing effects evoked by BMP-2 and strengthened fibrotic response. Exogenous BMP-2 supply could attenuate TGF-β1 signaling through Smad6-Smurf-1 complex activation. In vitro, mechanical stretch upregulated cardiac TGF-β1 and TGF-β1-dependent ROCK. While BMP-2 level was upregulated after blocking TGF-β1 signaling by SB-431542 or inhibition of ROCK by Y-27632. Dose-dependent TGF-β1 induction could activate ROCK and suppress endogenous BMP-2 level in cardiomyocytes. Knock-down BMP-2 in cardiomyocytes enhanced TGF-β1-mediated PKC-δand Smad3 signaling cascades. In contrast, application of Y-27632 or SB-431542 suppressed TGF-β1 pathway, but BMP-2 was only upregulated by Y-27632. BMP-2 silencing abolished Y-27632 but not SB-431542 mediated suppression of TGF-β1 pathway. Further experiments showed that the inhibitory Smad6 and Smurf1 were required for BMP-2-evoked antagonizing effects. Smad6 overexpression attenuated TGF-β1-induced activation of PKC-δand Smad3, promoted TGF-β RI degradation in BMP-2 knock-down cardiomyocytes Smad6 or Smurf1 siRNA could abolish the antagonizing effect of BMP-2 on TGF-β1 pathway, in which Smad6/Smurf1 complex formation was critically involved. In vivo data showed that pressure overload-induced collagen deposition were attenuated, cardiac function were improved and TGF-β1-dependent activation of PKC-δand Smad3 were reduced after 2 weeks treatment with rhBMP-2(0.5mg/kg) or Y-27632 (10mg/kg) in mice underwent surgical transverse aortic constriction. In conclusion, we propose that BMP-2, as a novel fibrosis antagonizing cytokine, may have potential beneficial effect in attenuating pressure overload-induced cardiac fibrosis. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 28 Jan 2012 | 4:03 pm CET

Semireplication-competent vesicular stomatitis virus as a novel platform for oncolytic virotherapy Abstract   Among oncolytic viruses, the vesicular stomatitis virus (VSV) is especially potent and a highly promising agent for the treatment of cancer. But, even though effective against multiple tumor entities in preclinical animal models, replication-competent VSV exhibits inherent neurovirulence, which has so far hindered clinical development. To overcome this limitation, replication-defective VSV vectors for cancer gene therapy have been tested and proven to be safe. However, gene delivery was inefficient and only minor antitumor efficacy was observed. Here, we present semireplication-competent vector systems for VSV (srVSV), composed of two trans-complementing, propagation-deficient VSV vectors. The de novo generated deletion mutants of the two VSV polymerase proteins P (phosphoprotein) and L (large catalytic subunit), VSVΔP and VSVΔL respectively, were used mutually or in combination with VSVΔG vectors. These srVSV systems copropagated in vitro and in vivo without recombinatory reversion to replication-competent virus. The srVSV systems were highly lytic for human glioblastoma cell lines, spheroids, and subcutaneous xenografts. Especially the combination of VSVΔG/VSVΔL vectors was as potent as wild-type VSV (VSV-WT) in vitro and induced long-term tumor regression in vivo without any associated adverse effects. In contrast, 90% of VSV-WT-treated animals succumbed to neurological disease shortly after tumor clearance. Most importantly, even when injected into the brain, VSVΔG/VSVΔL did not show any neurotoxicity. In conclusion, srVSV is a promising platform for virotherapeutic approaches and also for VSV-based vector vaccines, combining improved safety with an increased coding capacity for therapeutic transgenes, potentially allowing for multipronged approaches. Content Type Journal Article Category Original Article Pages 1-12 DOI 10.1007/s00109-012-0863-6 Authors Alexander Muik, Georg-Speyer-Haus, 60596 Frankfurt am Main, Germany Catherine Dold, Institute for Virology, Innsbruck Medical University, Fritz-Pregl-Str. 3, A-6020 Innsbruck, Austria Yvonne Geiß, Georg-Speyer-Haus, 60596 Frankfurt am Main, Germany Andreas Volk, Georg-Speyer-Haus, 60596 Frankfurt am Main, Germany Marina Werbizki, Georg-Speyer-Haus, 60596 Frankfurt am Main, Germany Ursula Dietrich, Georg-Speyer-Haus, 60596 Frankfurt am Main, Germany Dorothee von Laer, Institute for Virology, Innsbruck Medical University, Fritz-Pregl-Str. 3, A-6020 Innsbruck, Austria Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 27 Jan 2012 | 6:53 pm CET

New Paper in Press Microglia Stimulation of Glioblastoma Invasion Quelle: Molecular Medicine | 27 Jan 2012 | 6:51 pm CET

Elevated expression of cav-1 in a subset of SSc fibroblasts contributes to constitutive Alk1/Smad1 activation Abstract Previous studies have shown that the TGFβ/Alk1/Smad1 signaling pathway is constitutively activated in a subset of systemic sclerosis (SSc) fibroblasts and this pathway is a critical regulator of CCN2 gene expression. Caveolin-1 (cav-1), an integral membrane protein and the main component of caveolae, has also been implicated in SSc pathogenesis. This study was undertaken to evaluate the role of caveolin-1 in Smad1 signaling and CCN2 expression in healthy and SSc dermal fibroblasts. We show that a significant subset of SSc dermal fibroblasts have up-regulated cav-1 expression in vitro, and that cav-1 upregulation correlates with constitutive Smad1 phosphorylation. Additionally, basal levels of phospho-Smad1 were downregulated after inhibition of cav-1 in SSc dermal fibroblasts. Cav-1 formed a protein complex with Alk1 in dermal fibroblasts, and this association was enhanced by TGFβ. By using siRNA against cav-1 and adenoviral cav-1 overexpression we demonstrate that activation of Smad1 in response to TGFβ requires cav-1 and that cav-1 is sufficient for Smad-1 phosphorylation. We also show that cav-1 is a positive regulator of CCN2 gene expression, and that it is required for the basal and TGFβ-induced CCN2 levels. In conclusion, this study has revealed an important role of cav-1 in mediating TGFβ/Smad1 signaling and CCN2 gene expression in healthy and SSc dermal fibroblasts. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 25 Jan 2012 | 5:38 pm CET

Developmental regulation of inflammatory cytokine-mediated Stat3 signaling: the missing link between intrauterine growth restriction and pulmonary dysfunction? Abstract   Intrauterine growth restriction (IUGR) is a risk factor for impairment of lung function in adolescence and adulthood. Inflammatory and proliferative processes linking IUGR and perturbed extracellular matrix (ECM) as an underlying mechanism have not been addressed so far. Therefore, in this study, we aimed to investigate the developmental regulation of inflammatory and profibrotic processes in the lung subsequent to IUGR. IUGR was induced in rats by isocaloric protein restriction during gestation. Lung function was assessed with direct plethysmography at postnatal day (P) 28 and P70. Lungs were obtained at P1, P42, and P70 for assessment of mRNA, protein expression, immunohistochemistry, and gelatinolytic activity. Both respiratory system resistance and compliance were impaired subsequent to IUGR at P28 and this impairment was even more pronounced at P70. In line with these results, the expression of ECM components and metabolizing enzymes was deregulated. The deposition of collagen was increased at P70. In addition, the expression of inflammatory cytokines and both the activity and the expression of target genes of Stat3 signaling were dynamically regulated, with unaltered or decreased expression at P1 and significantly increased expression art P70. Taken together, these data give evidence for an age-dependent impairment of lung function as a result of a developmentally regulated increase in inflammatory and profibrotic processes subsequent to IUGR. Content Type Journal Article Category Original Article Pages 1-13 DOI 10.1007/s00109-012-0860-9 Authors Miguel Angel Alejandre Alcazar, Department of Pediatrics and Adolescent Medicine, University of Cologne, Kerpener Strasse 62, 50937 Cologne, Germany Iris Östreicher, Department of Neonatology, Charité University Medical Center, Berlin, Germany Sarah Appel, Department of Pediatrics and Adolescent Medicine, University of Cologne, Kerpener Strasse 62, 50937 Cologne, Germany Eva Rother, Department of Pediatrics and Adolescent Medicine, University of Cologne, Kerpener Strasse 62, 50937 Cologne, Germany Christina Vohlen, Department of Pediatrics and Adolescent Medicine, University of Cologne, Kerpener Strasse 62, 50937 Cologne, Germany Christian Plank, Department of Pediatrics and Adolescent Medicine, University of Erlangen—Nuremberg, Nuremberg, Germany Jörg Dötsch, Department of Pediatrics and Adolescent Medicine, University of Cologne, Kerpener Strasse 62, 50937 Cologne, Germany Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 24 Jan 2012 | 7:54 am CET

Evolutionary medicine and chronic inflammatory state—known and new concepts in pathophysiology Abstract   During the last 10 years, a series of exciting observations has led to a new theory of pathophysiology using insights from evolutionary biology and neuroendocrine immunology to understand the sequelae of chronic inflammatory disease. According to this theory, disease sequelae can be explained based on redirection of energy-rich fuels from storage organs to the activated immune system. These disease sequelae are highly diverse and include the following: sickness behavior, anorexia, malnutrition, muscle wasting–cachexia, cachectic obesity, insulin resistance with hyperinsulinemia, dyslipidemia, increase of adipose tissue near inflamed tissue, alterations of steroid hormone axes, elevated sympathetic tone and local sympathetic nerve fiber loss, decreased parasympathetic tone, hypertension, inflammation-related anemia, and osteopenia. Since these disease sequelae can be found in many animal models of chronic inflammatory diseases with mammals (e.g., monkeys, mice, rats, rabbits, etc.), the evolutionary time line goes back at least 70 million years. While the initial version of this theory could explain prominent sequelae of chronic inflammatory disease, it did not however address two features important in the pathogenesis of immune-mediated diseases: the time point when an acute inflammatory disease becomes chronic, and the appearance of hypertension in chronic inflammation. To address these aspects more specifically, a new version of the theory has been developed. This version defines more precisely the moment of transition from acute inflammatory disease to chronic inflammatory disease as a time in which energy stores become empty (complete energy consumption). Depending on the amount of stored energy, this time point can be calculated to be 19–43 days. Second, the revised theory addresses the mechanisms of essential hypertension since, on the basis of water loss, acute inflammatory diseases can stimulate water retention using a positively selected water retention system (identical to the energy provision system). In chronic smoldering inflammation, however, there is no increased water loss. In contrast, there is increased water generation in inflamed tissue and inflammatory cells, and the activation of the water retention system persists. This combination leads to a net increase of the systemic fluid volume, which is hypothesized to be the basis of essential hypertension (prevalence in adults 22–32%). Content Type Journal Article Category Review Pages 1-12 DOI 10.1007/s00109-012-0861-8 Authors Rainer H. Straub, Laboratory of Experimental Rheumatology and Neuroendocrino-Immunology, Division of Rheumatology, Department of Internal Medicine I, University Hospital Regensburg, 93042 Regensburg, Germany Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 24 Jan 2012 | 7:54 am CET

New Paper in Press BIN1 in hepatocellular carcinoma Quelle: Molecular Medicine | 23 Jan 2012 | 5:38 pm CET

Beneficial effects of edaravone, a novel antioxidant, in rats with dilated cardiomyopathy Abstract Edaravone, a novel antioxidant acts by trapping hydroxyl radicals, quenching active oxygen and so on. Its cardioprotective activity against experimental autoimmune myocarditis (EAM) was reported. Nevertheless, it remains to be determined whether edaravone protects against cardiac remodeling in dilated cardiomyopathy (DCM). The present study was undertaken to assess whether edaravone attenuates myocardial fibrosis and examine the effect of edaravone on cardiac function in rats with DCM after EAM. Rat model of EAM was prepared by injection with porcine cardiac myosin. 28 days after immunization, we administered edaravone intraperitoneally at 3 and 10 mg/kg/day to rats for 28 days. The results were compared with vehicle treated rats with DCM. Cardiac function, by hemodynamic and echocardiographic study and histopathology were performed. Left ventricular (LV) expression of NADPH oxidase subunits (p47phox, p67phox, gp91phox and Nox4), fibrosis markers (TGF-β1 and OPN), endoplasmic reticulum (ER) stress markers (GRP78 and GADD 153) and apoptosis markers (cytochrome C and caspase-3) were measured by Western blotting. Edaravone treated DCM rats showed better cardiac function compared with those of the vehicle treated rats. In addition, LV expressions of NADPH oxidase subunits levels were significantly down-regulated in edaravone-treated rats. Furthermore, the number of collagen-III positive cells in the myocardium of edaravone treated rats was lower compared with those of the vehicle treated rats. Our results suggest that edaravone ameliorated the progression of DCM by modulating oxidative and ER stress mediated myocardial apoptosis and fibrosis. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 23 Jan 2012 | 12:28 pm CET

Probing the polygenic basis of cardiomyopathies in Drosophila. Abstract In trying to understand the causes for congenital heart disease and cardiomyopathies, it is difficult to study polygenic interactions that contribute to the severity of the disease, which is in part due to genetic complexity and generation time of higher organsism that hinder efficient screening for modifiers of primary causes of heart disease. The adult Drosophila heart has recently been established as a model to probe genetic interactions that lead to cardiac dysfunction in this genetically simple and short-lived organism. This has made it possible to systematically and efficiently screen for polygenic modulators of heart dysfunction inflicted by known heart disease genes. Because heart development and fundamental aspects of cardiac physiology show remarkable evolutionary conservation, it has become possible to uncover new heart disease candidates by using Drosophila genetic tools in combination with sensitive heart function assays. Here, we review the discovery of several new genes, genetic pathways, and interactions that will help understanding human heart disease. For example, interactions between cardiogenic transcription factors, discovered in Drosophila, are also critical for adult heart function in flies and mammals. These include interactions between tinman/Nkx2-5 and neuromancer/Tbx20, which led to the discovery of possibly disease-causing familial variants in human TBX20. A new genetic pathway from tinman/Nkx2-5 to Cdc42, involving the microRNA miR-1, recently discovered in flies and subsequently validated to function similarly in mouse heart. Thus, the fly heart has proven to be a useful discovery tool for screening genetic interactions that are otherwise difficult to conduct. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 23 Jan 2012 | 12:20 pm CET

MicroRNA-24 Regulates Cardiac Fibrosis after Myocardial Infarction Abstract Cardiac fibrosis after myocardial infarction (MI) has been identified as a key factor in the development of heart failure. Although dysregulation of miRNAs is involved in various pathophysiologic processes in the heart, the role of miRNA in fibrosis regulation after MI is unclear. Previously we observed the correlation between fibrosis and the miR-24 expression in hypertrophic hearts, here we assessed how miR-24 regulates fibrosis after MI. Using qRT-PCR, we showed that microRNA-24 was down-regulated in the MI heart, the change in miR-24 expression was closely related to extracellular matrix (ECM) remodeling. In vivo, miR-24 could improve heart function and attenuate fibrosis in the infarct border zone of the heart 2 weeks after MI through intramyocardial injection of Lentiviruses. Moreover, in-vitro experiments suggested that up-regulation of miR-24 by synthetic miR-24 precursors could reduce fibrosis and also decrease the differentiation and migration of cardiac fibroblasts(CFs). TGF-β (a pathologic mediator of fibrotic disease) increased miR-24 expression, over-expression of miR-24 reduced TGF-β secretion and Smad2/3 phosphorylation in cardiac fibroblasts. Using microarray analyses and bioinformatics analyses, we found furin to be a potential target for miR-24 in fibrosis (furin is a protease which control latent TGF-β activation processing). Finally, we demonstrated that protein and mRNA levels of furin were regulated by miR-24 in cardiac fibroblasts. These findings suggest that miR-24 has a critical role in cardiac fibroblast function and cardiac fibrosis after MI through a furin–TGF-β pathway. Thus, miR-24 may be used as a target for treatment of MI and other fibrotic heart diseases. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 20 Jan 2012 | 5:39 am CET

Analysis of molecular mechanisms and anti-tumoral effects of zoledronic acid in breast cancer cells. Abstract Zoledronic acid (ZOL) is the most potent nitrogen-containing bisphosphonate (N-BPs) that strongly binds to bone mineral and acts as a powerful inhibitor of bone resorption, already clinically available for the treatment of patients with osteolytic metastases.Recent data also suggest that ZOL, used in breast cancer, may provide more than just supportive care modifying the course of the disease, though the possible molecular mechanism of action is still unclear. Since breast cancer is one of the primary tumors with high propensity to metastasize to the bone, we investigated, for the first time, differential gene expression profile on MCF-7 breast cancer cells treated with low doses of ZOL (10μM). Microarrays analysis was used to identify, describe and summarize evidences regarding the molecular basis of actions of ZOL and of their possible direct anti-tumor effects. We validated gene expression results of specific transcripts involved on major cellular process by Real Time and Western Blot analysis and we observed inhibition of proliferation and migration through MTT and Matrigel assay. We then focused on changes in the cytoskeletal components as FN1, actin, and anti angiogenic compounds as TGF-β1 and THBS1. The up-regulation of these products may have an important role in inhibiting proliferation, invasion and angiogenesis mediated by ZOL. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 20 Jan 2012 | 1:29 am CET

Amniotic Membrane-Derived Cells Inhibit Proliferation of Cancer Cell Lines by Inducing Cell Cycle Arrest Abstract Cells derived from the amniotic fetal membrane of human term placenta have drawn particular attention mainly for their plasticity and immunological properties, which render them interesting for stem cell research and cell-based therapeutic applications. In particular, we have previously demonstrated that amniotic mesenchymal tissue cells (AMTC) inhibit lymphocyte proliferation in vitro and suppress the generation and maturation of monocyte-derived dendritic cells. Here we show that AMTC also significantly reduce the proliferation of cancer cell lines of hematopoietic and non-hematopoietic origin, in both cell-cell contact and transwell co-cultures, therefore suggesting the involvement of yet unknown inhibitory soluble factor(s) in this “cell growth restraint”. Importantly, we provide evidence that the anti-proliferative effect of AMTC is associated to induction of cell cycle arrest in G0/G1 phase. Gene expression analyses demonstrate that AMTC can down-regulate cancer cells’ mRNA expression of genes associated with cell cycle progression, such as cyclins (cyclin D2, cyclin E1, cyclin H) and cyclin-dependent kinase (CDK4, CDK6 and CDK2), whilst they up-regulate cell cycle negative regulator such as p15 and p21, consistent with a block in G0/G1 phase with no progression to S phase. Taken together, these findings warrant further studies to investigate the applicability of these cells for controlling cancer cell proliferation in vivo. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 20 Jan 2012 | 1:28 am CET

Ethanol Increases Phosphate-Mediated Mineralization and Osteoblastic Transformation of Vascular Smooth Muscle Cells Abstract Vascular calcification is implicated in the pathogenesis of atherosclerosis, diabetes, and chronic kidney disease. Human vascular smooth muscle cells (HSMCs) undergo mineralization in response to elevated levels of inorganic phosphate (Pi) in an active and well-regulated process. This process involves increased activity of alkaline phosphatase and increased expression of core binding factor α-1 (CBF-α1), a bone-specific transcription factor, with the subsequent induction of osteocalcin. It has been shown that heavy alcohol consumption is associated with greater calcification in coronary arteries. The goal of our study was to examine whether ethanol alters mineralization of HSMCs provoked by high Pi. Exposure of HSMCs to ethanol increased extracellular matrix calcification in a dose responsive manner, providing a significant additional calcium deposition at concentrations of ≥60 mmol/L. HSMC calcification was accompanied by further enhancement in alkaline phosphatase activity. Ethanol also provoked a significant increase in the synthesis of osteocalcin. Moreover, in cells challenged with ethanol the expression of CBF-α1, a transcription factor involved in the regulation of osteoblastic transformation of HSMCs, was elevated. The observed effects of ethanol were not due to alterations of phosphate uptake by HSMCs. We conclude that ethanol enhances Pi-mediated human vascular smooth muscle calcification and transition of these cells into osteoblast-like cells. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 20 Jan 2012 | 1:26 am CET

Glucose can promote a glucocorticoid resistance state Abstract It has been shown that ingestion of glucose, amino acids, protein or mixed meals tends to increase serum and salivary cortisol concentrations in healthy adults. Recently, it has been demonstrated that morning glucose ingestion stimulates pulsatile cortisol and adrenocorticotropic hormone (ACTH) secretion, thus elevating their mean concentrations. In light of the above, a question arises: could the frequent food –and specifically glucose– consumption lead to hypercortisolism with possible clinical implications? And can the human body, under normal conditions raise defense mechanisms against the transient hypercortisolism caused by the frequent glucose consumption? Studies have revealed novel mechanisms which are implicated in the glucocorticoid receptor (GR)-mediated action, providing a kind of glucocorticoid resistance. This glucocorticoid resistance could be mediated through both enhancing acetylation (via, among others, regulation of essential clock genes such as Per) and inhibiting deacetylation of GR (via possible regulation of sirtuin activity). Interestingly, the acetylation/deacetylation processes seem to be regulated by glucose. Thus, glucose apart from causing increased cortisol secretion can, simultaneously, counter-regulate this hypercortisolism, by promoting directly and/or indirectly a glucocorticoid resistance state. Undoubtedly, before extracting conclusions regarding the clinical significance of the increased cortisol secretion following glucose ingestion, we should first thoroughly investigate the “defense” mechanisms provided by ‘nature’ to handle this hypercortisolism. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 20 Jan 2012 | 1:26 am CET

Human cardiomyocyte progenitor cells: a short history of nearly everything Quelle: Journal of Cellular and Molecular Medicine | 20 Jan 2012 | 1:25 am CET

Nano-regenerative medicine towards clinical outcome of stem cell and tissue engineering in humans Abstract Nanotechnology is a fast growing area of research that aims to create nanomaterials or nanostructures development in stem cell and tissue based therapies. Concepts and discoveries from the fields of bio nano research provide exciting opportunities of using stem cells for regeneration of tissues and organs. The application of nanotechnology to stem cell biology would be able to address the challenges of disease therapeutics. This review covers the potential of nanotechnology approaches towards regenerative medicine. Furthermore, it focuses on current aspects of stem and tissue cell engineering. The magnetic nanoparticles-based applications in stem cell research open new frontiers in cell and tissue engineering. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 20 Jan 2012 | 1:25 am CET

New Paper in Press miR-184 and miR-204 in Secondary Cataract Quelle: Molecular Medicine | 18 Jan 2012 | 3:24 pm CET

High expression of GCLC is associated with malignant melanoma of low oxidative phenotype and predicts a better prognosis Abstract   Reactive oxygen species (ROS) are strongly implicated in melanoma development, and treatment with antioxidants has shown efficacy in suppressing malignant transition and progression. Here, we investigated the significance of the glutamate-l-cysteine ligase catalytic subunit (GCLC) expression, a key regulator of glutathione synthesis, for malignant melanoma. A large set of melanoma cell lines (n = 36) was analyzed, and higher GCLC levels were associated with lower presence of intracellular ROS and interestingly also lower rates of cell proliferation. Moreover, treatment with the antioxidant N-acetylcysteine efficiently reduced the growth speed of several investigated malignant cells. In addition GCLC expression was significantly linked to a prominent set of cellular antioxidants, accounting for the observed lower basal levels of oxidative stress and higher antioxidative capacity. Key attributes defining the malignant phenotype of melanoma cells including survival, invasiveness, and switch from E-cadherin to N-cadherin expression were more prominent in cells with lower GCLC expression. Our findings were further corroborated by observations in Rag2−/−γc−/−mice, in which melanoma cells with lower GCLC expression depicted a dramatically stronger tumor growth. Furthermore, prognostic significance of GCLC expression was investigated in patients (n = 28) with advanced malignant melanoma. High tumor immunoreactivity for GCLC was a significant determinant for better 5-year overall survival. Conclusively, we show for the first time that GCLC may serve a dual role, as a surrogate marker for cellular redox state as well as malignant potential of melanoma cells. These promising results regarding its prognostic significance as well as its potential as a pharmacological target require further in-depth investigations. Content Type Journal Article Category Original Article Pages 1-10 DOI 10.1007/s00109-012-0857-4 Authors Dimitrios Mougiakakos, Department of Oncology and Pathology, Cancer Center Karolinska, Karolinska Institutet, Stockholm, Sweden Riki Okita, Department of Oncology and Pathology, Cancer Center Karolinska, Karolinska Institutet, Stockholm, Sweden Takashi Ando, Department of Orthopaedic Surgery, University of Yamanashi, Yamanashi, 409-3898 Japan Christoph Dürr, Division of Hematology and Oncology, Department of Medicine, Freiburg University Medical Center, Albert-Ludwigs-University, Freiburg, Germany Jules Gadiot, Department of Medical Oncology and Division of Immunology, The Netherlands Cancer Institute-Antoni van Leeuwenhoek Hospital (NKI-AVL), Amsterdam, 1066 CX The Netherlands Jiro Ichikawa, Department of Orthopaedics and Pharmacology, Vanderbilt University, Nashville, TN, USA Robert Zeiser, Division of Hematology and Oncology, Department of Medicine, Freiburg University Medical Center, Albert-Ludwigs-University, Freiburg, Germany Christian Blank, Department of Medical Oncology and Division of Immunology, The Netherlands Cancer Institute-Antoni van Leeuwenhoek Hospital (NKI-AVL), Amsterdam, 1066 CX The Netherlands C. Christian Johansson, Department of Oncology and Pathology, Cancer Center Karolinska, Karolinska Institutet, Stockholm, Sweden Rolf Kiessling, Department of Oncology and Pathology, Cancer Center Karolinska, Karolinska Institutet, Stockholm, Sweden Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 17 Jan 2012 | 8:09 am CET

The non-anticoagulant heparin-like K5 polysaccharide derivative K5-N,OSepi attenuates myocardial ischemia/reperfusion injury Abstract Heparin and low molecular weight heparins have been demonstrated to reduce myocardial ischemia/reperfusion (I/R) injury, although their use is hampered by the risk of hemorrhagic and thrombotic complications. Chemical and enzymatic modifications of K5 polysaccharide have shown the possibility to produce heparin-like compounds with low anticoagulant activity and strong anti-inflammatory effects. Using a rat model of regional myocardial I/R, we investigated the effects of an epimerized N-,O-sulfated K5 polysaccharide derivative, K5-N,OSepi, on infarct size and histological signs of myocardial injury caused by 30 min ligature of the left anterior descending coronary artery followed by 1 or 24 h reperfusion. K5-N,OSepi (0.1-1 mg/kg given i.v. 15 min before reperfusion) significantly reduced the extent of myocardial damage in a dose-dependent manner. Furthermore, we investigated the potential mechanism(s) of the cardioprotective effect(s) afforded by K5-N,OSepi. In left ventricular samples, I/R induced mast cell degranulation and a robust increase in lipid peroxidation, free radical-induced DNA damage and calcium overload. Markers of neutrophil infiltration and activation were also induced by I/R in rat hearts, specifically myeloperoxidase activity, intercellular-adhesion-molecule-1 expression, prostaglandin-E2 and tumor-necrosis-factor-α production. The robust increase in oxidative stress and inflammatory markers was blunted by K5-N,OSepi, in a dose-dependent manner, with maximum at 1 mg/kg. Furthermore, K5-N,OSepi administration attenuated the increase in caspase 3 activity, Bid and Bax activation and ameliorated the decrease in expression of Bcl-2 within the ischemic myocardium. In conclusion, we demonstrate that the cardioprotective effect of the non-anticoagulant K5 derivative K5-N,OSepi is secondary to a combination of anti-apoptotic and anti-inflammatory effects. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 17 Jan 2012 | 3:12 am CET

Cardiac Differentiation of Human Pluripotent Stem Cells Abstract Due to the extremely limited proliferative capacity of adult cardiomyocytes, human embryonic (pluripotent) stem cell derived cardiomyocytes (hESC-CMs) are currently almost the only reliable source of human heart cells which are suited to large-scale production. These cells have the potential for wide-scale application in drug discovery, heart disease research and cell-based heart repair. Embryonic atrial-, ventricular- and nodal-like cardiomyocytes can be obtained from differentiated human embryonic stem cells (hESCs). In recent years, several highly-efficient cardiac differentiation protocols have been developed. Significant progress has also been made on understanding cardiac subtype specification, which is the key to reducing the heterogeneity of hESC-CMs, a major obstacle to the utilization of these cells in medical research and future cell-based replacement therapies. Here we review recent progress in cardiac differentiation of hESCs and cardiac subtype specification, and discuss potential applications in drug screening and cell-based heart regeneration. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 17 Jan 2012 | 12:37 am CET

Angiotensin converting enzyme 2 abrogates bleomycin-induced lung injury Abstract   Despite substantial progress, mortality and morbidity of the acute respiratory distress syndrome (ARDS), a severe form of acute lung injury (ALI), remain unacceptably high. There is no effective treatment for ARDS/ALI. The renin–angiotensin system (RAS) through Angiotensin-converting enzyme (ACE)-generated Angiotensin II contributes to lung injury. ACE2, a recently discovered ACE homologue, acts as a negative regulator of the RAS and counterbalances the function of ACE. We hypothesized that ACE2 prevents Bleomycin (BLM)-induced lung injury. Fourteen to 16-week-old ACE2 knockout mice—male (ACE2−/y) and female (ACE2−/−)—and age-matched wild-type (WT) male mice received intratracheal BLM (1.5U/kg). Male ACE2−/y BLM injured mice exhibited poorer exercise capacity, worse lung function and exacerbated lung fibrosis and collagen deposition compared with WT. These changes were associated with increased expression of the profibrotic genes α-smooth muscle actin (α-SMA) and Transforming Growth Factor ß1. Compared with ACE2−/y exposed to BLM, ACE2−/− exhibited better lung function and architecture and decreased collagen deposition. Treatment with intraperitoneal recombinant human (rh) ACE2 (2 mg/kg) for 21 days improved survival, exercise capacity, and lung function and decreased lung inflammation and fibrosis in male BLM-WT mice. Female BLM WT mice had mild fibrosis and displayed a possible compensatory upregulation of the AT2 receptor. We conclude that ACE2 gene deletion worsens BLM-induced lung injury and more so in males than females. Conversely, ACE2 protects against BLM-induced fibrosis. rhACE2 may have therapeutic potential to attenuate respiratory morbidity in ALI/ARDS. Content Type Journal Article Category Original Article Pages 1-11 DOI 10.1007/s00109-012-0859-2 Authors G. J. Rey-Parra, Department of Pediatrics, Women and Children Health Research Institute, Cardiovascular Research Center, Pulmonary Research Group, University of Alberta, Edmonton, Canada A. Vadivel, Department of Pediatrics, Women and Children Health Research Institute, Cardiovascular Research Center, Pulmonary Research Group, University of Alberta, Edmonton, Canada L. Coltan, Department of Pediatrics, Women and Children Health Research Institute, Cardiovascular Research Center, Pulmonary Research Group, University of Alberta, Edmonton, Canada A. Hall, Department of Pediatrics, Women and Children Health Research Institute, Cardiovascular Research Center, Pulmonary Research Group, University of Alberta, Edmonton, Canada F. Eaton, Department of Pediatrics, Women and Children Health Research Institute, Cardiovascular Research Center, Pulmonary Research Group, University of Alberta, Edmonton, Canada M. Schuster, Apeiron Biologics, Vienna, Austria H. Loibner, Apeiron Biologics, Vienna, Austria J. M. Penninger, IMBA, Vienna, Austria Z. Kassiri, Department of Physiology, University of Alberta, Edmonton, Canada G. Y. Oudit, Department of Physiology, University of Alberta, Edmonton, Canada B. Thébaud, Department of Pediatrics, Women and Children Health Research Institute, Cardiovascular Research Center, Pulmonary Research Group, University of Alberta, Edmonton, Canada Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 13 Jan 2012 | 5:50 pm CET

Non-hypoxic stabilization of HIF-Iα during coordinated interaction between Akt and angiopoietin-1 enhances endothelial commitment of bone marrow stem cells Abstract   We previously reported that mesenchymal stem cells (MSC) co-expressing Akt and angiopoietin-1 (Ang-1) preserved infarcted heart function via angiomyogenesis. The present study determined the mechanism of co-overexpression of Akt and Ang-1 in promoting endothelial commitment of MSC. The cells were transduced with vectors encoding for Akt (AktMSC), Ang-1 (Ang-1MSC), and both Akt and Ang-1 (AAMSC) using Empty vector transduced MSC (EmpMSC) as control. Molecular studies indicated a coordinated interaction between Akt and Ang-1 in AAMSC and led to non-hypoxic stabilization of hypoxia inducible factor-1α (HIF-Iα) which accentuated under 4-h anoxia. We also observed HIF-Iα dependent induction of hemeoxygenase-1, endothelial specific markers and VEGF in AAMSC. Vascular commitment of AAMSC was confirmed by immunostaining, Western blotting and flow cytometry for endothelial specific early and late markers including Flt1, Flk1, Tie2, VCAM-1, and von Willebrand Factor-VIII (vWF-VIII) in HIF-Iα dependent fashion besides exhibiting higher emigrational activity and angiogenesis in vitro. AAMSC transplanted into rat model of myocardial infarction showed higher Flk1 and Flt1 positivity and also promoted intrinsic Flk1+ and Flt1+ cell mobilization into the infarcted heart. Given the ease of availability of MSC and simplicity of approach to co-overexpress Ang-1 and Akt to enhance their endothelial commitment, the strategy will be significant for cellular angiogenesis to treat ischemic heart. Content Type Journal Article Category Original Article Pages 1-12 DOI 10.1007/s00109-011-0852-1 Authors Vien Khach Lai, Department of Pathology, University of Cincinnati Medical Center, 231-Albert Sabin Way, Cincinnati, OH 45267-0529, USA Muhammad Rizwan Afzal, Department of Pathology, University of Cincinnati Medical Center, 231-Albert Sabin Way, Cincinnati, OH 45267-0529, USA Muhammad Ashraf, Department of Pathology, University of Cincinnati Medical Center, 231-Albert Sabin Way, Cincinnati, OH 45267-0529, USA Shujia Jiang, Department of Pathology, University of Cincinnati Medical Center, 231-Albert Sabin Way, Cincinnati, OH 45267-0529, USA Husnain Kh Haider, Department of Pathology, University of Cincinnati Medical Center, 231-Albert Sabin Way, Cincinnati, OH 45267-0529, USA Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 11 Jan 2012 | 6:54 pm CET

Inhibitors of hypoxia-inducible factor 1 block breast cancer metastatic niche formation and lung metastasis Abstract   Intratumoral hypoxia, a frequent finding in metastatic cancer, results in the activation of hypoxia-inducible factors (HIFs). HIFs are implicated in many steps of breast cancer metastasis, including metastatic niche formation through increased expression of lysyl oxidase (LOX) and lysyl oxidase-like (LOXL) proteins, enzymes that remodel collagen at the metastatic site and recruit bone marrow-derived cells (BMDCs) to the metastatic niche. We investigated the effect of two chemically and mechanistically distinct HIF inhibitors, digoxin and acriflavine, on breast cancer metastatic niche formation. Both drugs blocked the hypoxia-induced expression of LOX and LOXL proteins, collagen cross-linking, CD11b+ BMDC recruitment, and lung metastasis in an orthotopic breast cancer model. Patients with HIF-1α-overexpressing breast cancers are at increased risk of metastasis and mortality and our results suggest that such patients may benefit from aggressive therapy that includes a HIF inhibitor. Content Type Journal Article Category Original Article Pages 1-13 DOI 10.1007/s00109-011-0855-y Authors Carmen Chak-Lui Wong, Vascular Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA Huafeng Zhang, Vascular Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA Daniele M. Gilkes, Vascular Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA Jasper Chen, Vascular Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA Hong Wei, Vascular Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA Pallavi Chaturvedi, Vascular Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA Maimon E. Hubbi, Vascular Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA Gregg L. Semenza, Vascular Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 9 Jan 2012 | 8:37 pm CET

Retraction Note to: Direct renin inhibition: clinical pharmacology Retraction Note to: Direct renin inhibition: clinical pharmacology Content Type Journal Article Category Retraction Note Pages 1-1 DOI 10.1007/s00109-011-0850-3 Authors Michel Azizi, Faculté de Médecine, Université Paris Descartes; Assistance Publique Hôpitaux de Paris, Hôpital Européen Georges Pompidou; INSERM, Clinical Investigation Center 9201, Paris, France Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 9 Jan 2012 | 8:37 pm CET

A new, powerful player in lipoprotein metabolism: brown adipose tissue Abstract   Important causes for modern epidemics such as obesity, diabetes, and cardiovascular disease are over- and malnutrition. Dietary as well as endogenous lipids are transported through the bloodstream in lipoproteins, and disturbances in lipoprotein metabolism are associated with atherosclerosis, heart disease, and diabetes. Recent findings reveal biological principles—how lipoproteins, in particular triglyceride-rich lipoproteins, are metabolized and what factors regulate their processing. The fate of triglycerides delivered by lipoproteins is quite simple: either they can be stored or they can be utilized for combustion or biosynthetic pathways. In the healthy state, fatty acids derived from triglycerides can be burned in the heart, muscle, and other organs for actual work load, or they can be stored in white adipose tissue. The combination of storage and combustion is realized in brown adipose tissue (BAT), a peripheral organ that was long thought to be only of relevance in small mammals: Recent data however prove that BAT plays an important role in human adults. Here, we will review recent insights on how BAT controls triglyceride clearance and the possible implications for the treatment of chronic diseases caused by lipid mishandling. Content Type Journal Article Category Review Pages 1-7 DOI 10.1007/s00109-012-0858-3 Authors Alexander Bartelt, Department of Biochemistry and Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, Martinistraße 52, 20246 Hamburg, Germany Martin Merkel, Department of Biochemistry and Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, Martinistraße 52, 20246 Hamburg, Germany Joerg Heeren, Department of Biochemistry and Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, Martinistraße 52, 20246 Hamburg, Germany Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 9 Jan 2012 | 8:37 pm CET

IL-17 producing T cells in mouse models of multiple sclerosis and rheumatoid arthritis Abstract   Multiple Sclerosis (MS) and Rheumatoid Arthritis (RA) are amongst the most common autoimmune diseases in the northern hemisphere. There is mounting evidence that in both afflictions, not only environmental and genetic factors influence disease, but cellular components such as autoreactive T cells also contribute to pathology. Animal models are key in the study and subsequent therapeutic development for human autoimmune diseases. As patient material is often difficult to obtain and in some cases—as in MS, where the central nervous system (CNS) is concerned—even not accessible, animal models provide a multifaceted tool to explore disease-underlying mechanisms. The pro-inflammatory T cell cytokine IL-17 has recently moved to center stage due to its crucial role in autoimmune diseases including MS and RA. A plethora of studies in animal models has sustained the relevance of this cytokine pathway for the development of autoimmunity and shed light on its cellular sources and patho-mechanisms. This review addresses the role of IL-17 producing T lymphocytes, in particular CD4+ and γδ T cells, in three commonly used mouse models for MS and RA, namely experimental autoimmune encephalomyelitis (EAE), collagen-induced arthritis (CIA), and antigen-induced arthritis (AIA). Comparing and combining knowledge gained from different animal models will broaden our understanding of the IL-17 biology and facilitate the translation to the human diseases. Content Type Journal Article Category Review Pages 1-12 DOI 10.1007/s00109-011-0841-4 Authors Bernadette Pöllinger, Department of Autoimmunity, Transplantation & Inflammation, Novartis Institutes for BioMedical Research, Basel, Switzerland Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 9 Jan 2012 | 8:37 pm CET

Chemopreventive effect of the non-psychotropic phytocannabinoid cannabidiol on experimental colon cancer Abstract   Colon cancer affects millions of individuals in Western countries. Cannabidiol, a safe and non-psychotropic ingredient of Cannabis sativa, exerts pharmacological actions (antioxidant and intestinal antinflammatory) and mechanisms (inhibition of endocannabinoid enzymatic degradation) potentially beneficial for colon carcinogenesis. Thus, we investigated its possible chemopreventive effect in the model of colon cancer induced by azoxymethane (AOM) in mice. AOM treatment was associated with aberrant crypt foci (ACF, preneoplastic lesions), polyps, and tumour formation, up-regulation of phospho-Akt, iNOS and COX-2 and down-regulation of caspase-3. Cannabidiol-reduced ACF, polyps and tumours and counteracted AOM-induced phospho-Akt and caspase-3 changes. In colorectal carcinoma cell lines, cannabidiol protected DNA from oxidative damage, increased endocannabinoid levels and reduced cell proliferation in a CB1-, TRPV1- and PPARγ-antagonists sensitive manner. It is concluded that cannabidiol exerts chemopreventive effect in vivo and reduces cell proliferation through multiple mechanisms. Content Type Journal Article Category Original Article Pages 1-10 DOI 10.1007/s00109-011-0856-x Authors Gabriella Aviello, Department of Experimental Pharmacology, Endocannabinoid Research Group, University of Naples Federico II, Naples, Italy Barbara Romano, Department of Experimental Pharmacology, Endocannabinoid Research Group, University of Naples Federico II, Naples, Italy Francesca Borrelli, Department of Experimental Pharmacology, Endocannabinoid Research Group, University of Naples Federico II, Naples, Italy Raffaele Capasso, Department of Experimental Pharmacology, Endocannabinoid Research Group, University of Naples Federico II, Naples, Italy Laura Gallo, Institute of Biomolecular Chemistry, Endocannabinoid Research Group, Consiglio Nazionale delle Ricerche (CNR), Pozzuoli, Italy Fabiana Piscitelli, Institute of Biomolecular Chemistry, Endocannabinoid Research Group, Consiglio Nazionale delle Ricerche (CNR), Pozzuoli, Italy Vincenzo Di Marzo, Institute of Biomolecular Chemistry, Endocannabinoid Research Group, Consiglio Nazionale delle Ricerche (CNR), Pozzuoli, Italy Angelo A. Izzo, Department of Experimental Pharmacology, Endocannabinoid Research Group, University of Naples Federico II, Naples, Italy Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 9 Jan 2012 | 8:37 pm CET

LINE-1 hypomethylation in familial and sporadic cancer Abstract   Increased and decreased methylation at specific sequences (hypermethylation and hypomethylation, respectively) is characteristic of tumor DNA compared to normal DNA and promotes carcinogenesis in multiple ways including genomic instability. Long interspersed element (LINE), an abundant class of retrotransposons, provides a surrogate marker for global hypomethylation. We developed methylation-specific multiplex ligation-dependent probe amplification assays to study LINE-1 methylation in cases of colorectal, gastric, and endometrial cancer (N = 276), stratified by patient category [sporadic; Lynch syndrome (LS); familial colorectal cancer type X (FCCX)] and microsatellite instability status. Within each patient group, LINE-1 showed lower methylation in tumor DNA relative to paired normal DNA and hypomethylation was statistically significant in most cases. Interestingly, normal colorectal mucosa samples from different patient groups displayed differences in LINE-1 methylation that mirrored differences between the respective tumor tissues, with a decreasing trend for LINE-1 methylation from patients with sporadic colorectal cancer to LS to FCCX. Despite the fact that the degree of LINE-1 methylation is generally tissue specific, normal colorectal mucosa, gastric mucosa, and endometrium from LS patients showed similar levels of LINE-1 methylation. Our results suggest that the degree of LINE-1 methylation may constitute a “field defect” that may predispose normal tissues for cancer development. Content Type Journal Article Category Original Article Pages 1-9 DOI 10.1007/s00109-011-0854-z Authors Walter Pavicic, Department of Medical Genetics, Haartman Institute, University of Helsinki, Helsinki, PO Box 63, 00014 Finland Emmi I. Joensuu, Department of Medical Genetics, Haartman Institute, University of Helsinki, Helsinki, PO Box 63, 00014 Finland Taina Nieminen, Department of Medical Genetics, Haartman Institute, University of Helsinki, Helsinki, PO Box 63, 00014 Finland Päivi Peltomäki, Department of Medical Genetics, Haartman Institute, University of Helsinki, Helsinki, PO Box 63, 00014 Finland Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 7 Jan 2012 | 5:55 pm CET

Platelet derived growth factor receptor α-positive cells in the tunica muscularis of human colon Abstract Background and Aims: An obstacle to understanding motor pathologies of the GI tract is that the physiology of some of the cellular components of the gut wall is not understood. Morphologists identified fibroblast-like cells in the tunica muscularis many years ago, but little is know about these interstitial cells because of inadequate techniques to identify these cells. Recent findings have shown that fibroblast-like cells express PDGFRα in mice and antibodies for these receptors can be used to label the cells. Methods: We used immunohistochemical techniques to study the phenotype and intercellular relationships of fibroblast-like cells in the human colon. Results: Fibroblast-like cells are labeled specifically with antibodies to PDGFRα and widely distributed through the tunica muscularis of human colon. These cells form discrete networks in the region of the myenteric plexus and within the circular and longitudinal muscle layers. PDGFRα+ cells are distinct from c-Kit+ interstitial cells of Cajal and closely associated with varicose processes of neurons expressing substance P (excitatory motor neurons) or nNOS (inhibitory motor neurons). PDGFRα+ cells express small conductance Ca2+-activated K+ channels (SK3), which are likely to mediate purinergic neural regulation of colonic muscles. Conclusions: Our data suggest that PDGFRα+ cells may have an important role in transducing inputs from enteric motor neurons. This study identifies reagents and techniques that will allow investigation of this class of interstitial cells and help to develop an understanding of the role of PDGFRα+ cells in the human GI tract in health and disease. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 6 Jan 2012 | 10:55 pm CET

Raf-1 levels determine the migration rate of primary endometrial stromal cells of patients with endometriosis Abstract Endometriosis is a disease characterized by the localization of endometrial tissue outside of the uterine cavity. The differences observed in migration of human endometrial stromal cells (hESC) obtained from patients with endometriosis versus healthy controls were proposed to correlate with the abnormal activation of Raf-1/ROCKII signalling pathway. To evaluate the mechanism by which Raf-1 regulates cytoskeleton reorganization and motility, we used primary eutopic (Eu-, n=16) and ectopic (Ec-, n=8; isolated from ovarian cysts) hESC of patients with endometriosis and endometriosis-free controls (Co-hESC, n=14). Raf-1 siRNA knockdown in Co- and Eu-hESC resulted in contraction and decreased migration vs. siRNA controls. This phenotype was reversed following the re-expression of Raf-1 in these cells. Lowest Raf-1 levels in Ec-hESC were associated with hyperactivated ROCKII and ezrin/radixin/moesin (E/R/M), impaired migration and a contracted phenotype similar to Raf-1 knockdown in Co- and Eu-hESC. We further show that the mechanism by which Raf-1 mediates migration in hESC includes direct myosin light chain phosphatase (MYPT1) phosphorylation and regulation of the levels of E/R/M, paxillin, MYPT1 and myosin light chain (MLC) phosphorylation indirectly via the hyperactivation of ROCKII kinase. Further, we suggest that in contrast to Co-and Eu-hESC, where the cellular Raf-1 levels regulate the rate of migration, the low cellular Raf-1 content in Ec-hESC might ensure their restricted migration by preserving the contracted cellular phenotype. In conclusion, our findings suggest that cellular levels of Raf-1 adjust the threshold of hESC migration in endometriosis. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 6 Jan 2012 | 10:39 pm CET

A fibronectin-fibrinogen-tropoelastin coating reduces smooth muscle cell growth but improves endothelial cell function Abstract Reendothelialization of the stent surface after PCI is known to be an important determinant of clinical outcome. We compared the effects of biological stent coatings, fibronectin, fibrinogen and tropoelastin, on umbilical vein endothelial cell (HUVEC) and vascular smooth muscle cell (VSMC) characteristics. Umbilical cord arterial segments were cultured on coated surfaces and VSMC outgrowth (indicating proliferation and migration) was measured after 12 days. mRNA was isolated from HUVEC and VSMC cultured on these coatings and gene expression was profiled by QPCR. Procoagulant properties of HUVEC were determined by an indirect chromogenic assay which detects tissue factor activity. The varying stent coatings influence VSMC outgrowth: 31.2±4.0mm2 on fibronectin, 1.6±0.3mm2 on tropoelastin and 8.1±1.5mm2 on a mixture of fibronectin/fibrinogen/tropoelastin, while HUVEC migration remains unaffected. Culturing HUVEC on tropoelastin induces increased expression of VCAM-1 (13.1±4.4pg/ml), ICAM-1 (5.1±1.3pg/ml) and IL-8 (11.6±3.1pg/ml) compared to fibronectin (0.7±0.2pg/ml, 0.8±0.2pg/ml, 2.3±0.5pg/ml respectively), while expression levels on fibronectin/fibrinogen/tropoelastin remain unaltered. No significant differences in VCAM-1, ICAM-1 and IL-8 mRNA expression are found in VSMC. Finally, HUVEC cultured on tropoelastin display a 5-fold increased tissue factor activity (511.6±26.7%), compared to cells cultured on fibronectin (100%±3.9%) or fibronectin/fibrinogen/tropoelastin (76.3±25.0%). These results indicate that tropoelastin inhibits VSMC migration but leads to increased inflammatory and procoagulant markers on endothelial cells. Fibronectin/fibrinogen/tropoelastin inhibits VSMCs while compensating the inflammatory and procoagulant effects. These data suggest that coating a mixture of fibronectin/fibrinogen/tropoelastin on a stent may promote reendothelialization, while keeping unfavorable processes such as restenosis and procoagulant activity limited. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 6 Jan 2012 | 10:39 pm CET

Rhabdomyosarcomas: an overview on the experimental animal models Abstract Rhabdomyosarcomas (RMS) are aggressive childhood soft-tissue malignancies deriving from mesenchymal progenitors that are committed to muscle-specific lineages. Despite the histopathological signatures are associated to three main histological variants, termed embryonal, alveolar and pleomorphic, a plethora of genetical and molecular changes are recognized in RMS. Over the years, exposure to carcinogens or ionizing radiations and gene targeting approaches in vivo have greatly contributed to disclose some of the mechanisms underlying RMS onset. In this review, we describe the principal distinct features associated to RMS variants and focus on the current available experimental animal models to point out the molecular determinants cooperating to RMS development and progression. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 6 Jan 2012 | 10:38 pm CET

Measuring passive myocardial stiffness in Drosophila melanogaster to investigate diastolic dysfunction Aging is marked by a decline in left ventricular diastolic function, which encompasses abnormalities in diastolic relaxation, chamber filling and/or passive myocardial stiffness. Genetic tractability and short life span make Drosophila melanogaster an ideal organism to study the effects of aging on heart function, including senescent-associated changes in gene expression and in passive myocardial stiffness. However, use of the Drosophila heart tube to probe deterioration of diastolic performance is subject to at least two challenges: the extent of genetic homology to mammals and the ability to resolve mechanical properties of the bilayered fly heart, which consists of a ventral muscle layer that covers the contractile cardiomyocytes. Here we argue for wide-spread use of Drosophila as a novel myocardial aging model by 1) describing diastolic dysfunction in flies, 2) discussing how critical pathways involved in dysfunction are conserved across species, and 3) demonstrating the advantage of an atomic force microscopy-based analysis method to measure stiffness of the multilayered Drosophila heart tube versus isolated myocytes from other model systems. By using powerful Drosophila genetic tools we aim to efficiently alter changes observed in factors that contribute to diastolic dysfunction to understand how one might improve diastolic performance at advanced ages in humans. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 6 Jan 2012 | 10:38 pm CET

MicroRNA control of myelopoiesis and the differentiation block in Acute Myeloid Leukaemia Abstract In the relatively short period of time since their discovery microRNAs have been shown to control many important cellular functions such as cell differentiation, growth, proliferation and apoptosis. Additionally, microRNAs have been demonstrated as key drivers of many malignancies and can function as either tumour suppressors or oncogenes. The haematopoietic system is not outside the realm of microRNA control with microRNAs controlling aspects of stem cell and progenitor self-renewal and differentiation; with many, if not all haematological disorders associated with aberrant microRNA expression and function. In this review we focus on the current understanding of microRNA control of haematopoiesis and detail the evidence for the contribution and clinical relevance of aberrant microRNA function to the characteristic block of differentiation in Acute Myeloid Leukaemia. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 6 Jan 2012 | 10:38 pm CET

Dose-Dependent Functional Benefit of Human Cardiosphere Transplantation in Mice with Acute Myocardial Infarction Abstract Despite mounting preclinical and clinical evidence of the beneficial effects of cell-based therapy, optimal cell dosing and delivery approaches have not been identified. Cardiospheres are self-assembling 3D microtissues formed by cardiac stem cells and supporting cell types. The ability of cardiospheres to augment cardiac function has been demonstrated in animal models of ischemic cardiomyopathy. In the present study, we studied the dose-dependence of the benefits of human cardiospheres, delivered via intramyocardial injection, upon cardiac function and ventricular remodeling in SCID mice with acute myocardial infarction. Four doses of cardiospheres were used: 1x104, 5x104, 1x105, and 5x105 (expressed as number of plated cardiosphere-forming cells). Acute (24 hour) cell retention rates in all groups were similar. Functional assessment and quantitative heart morphometry indicated benefit from higher cell doses (>5x104) in terms of ejection fraction, infarct size and capillary density. Histological analysis indicated that the dose-dependent benefit was primarily due to indirect effects of transplanted cells. The results provide scalable data on cardiosphere dosing for intramyocardial injection. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 6 Jan 2012 | 10:38 pm CET

Anti-inflammatory mechanisms and therapeutic opportunities in myocardial infarct healing Abstract   The wound healing response after myocardial infarction (MI) involves a cascade of molecular and cellular events that lead to a replacement of the necrotic area with a collagen-rich scar. Clearance of necrotic debris by neutrophils, monocytes, and macrophages is a critical component of infarct healing; however, tight control and timely repression of this inflammatory response is important to prevent excessive tissue degradation leading to infarct expansion and heart failure. Genetic ablation or blockade of anti-inflammatory pathways tends to be detrimental after MI, whereas genetic ablation of pro-inflammatory pathways tends to be beneficial. Accordingly, therapies enhancing endogenous anti-inflammatory pathways or blocking endogenous pro-inflammatory pathways have been found to improve wound healing and to reduce the risk of heart failure in rodent models of acute MI. Besides their scavenger function, inflammatory cells promote healing by stimulating angiogenesis and granulation tissue formation via paracrine factors. Moreover, signaling mediators that are active in inflammatory cells may be active also in non-inflammatory cell types involved in infarct healing. Some anti-inflammatory interventions are therefore deleterious. However, interventions that carefully adjust the balance between the essential and detrimental facets of inflammation may provide new therapeutic opportunities for patients with large MIs who continue to be at risk of developing heart failure, despite modern reperfusion and anti-remodeling strategies. Content Type Journal Article Category Review Pages 1-9 DOI 10.1007/s00109-011-0847-y Authors Tibor Kempf, Division of Molecular and Translational Cardiology, Department of Cardiology and Angiology, Hannover Medical School, Hannover, Germany Alexander Zarbock, Department of Anesthesiology and Intensive Care Medicine, University of Münster, Münster, Germany Dietmar Vestweber, Max-Planck Institute for Molecular Biomedicine, Münster, Germany Kai C. Wollert, Division of Molecular and Translational Cardiology, Department of Cardiology and Angiology, Hannover Medical School, Hannover, Germany Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 6 Jan 2012 | 5:47 pm CET

Gastrin inhibits a novel, pathological colon cancer signaling pathway involving EGR1, AE2, and P-ERK Abstract   Human anion exchanger 2 (AE2) is a plasma membrane protein that regulates intracellular pH and cell volume. AE2 contributes to transepithelial transport of chloride and bicarbonate in normal colon and other epithelial tissues. We now report that AE2 overexpression in colon cancer cells is correlated with expression of the nuclear proliferation marker, Ki67. Survival analysis of 24 patients with colon cancer in early stage or 33 patients with tubular adenocarcinoma demonstrated that expression of AE2 is correlated with poor prognosis. Cellular and molecular experiments indicated that AE2 expression promoted proliferation of colon cancer cells. In addition, we found that transcription factor EGR1 underlies AE2 upregulation and the AE2 sequester p16INK4a (P16) in the cytoplasm of colon cancer cells. Cytoplasmic P16 enhanced ERK phosphorylation and promoted proliferation of colon cancer cells. Gastrin inhibited proliferation of colon cancer cells by suppressing expression of EGR1 and AE2 and by blocking ERK phosphorylation. Taken together, our data describe a novel EGR1/AE2/P16/P-ERK signaling pathway in colon carcinogenesis, with implications for pathologic prognosis and for novel therapeutic approaches. Content Type Journal Article Category Original Article Pages 1-12 DOI 10.1007/s00109-011-0851-2 Authors Ling-Jun Song, Department of Pathology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025 People’s Republic of China Rui-Jun Liu, Department of Pathology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025 People’s Republic of China Zhi Zeng, Department of Pathology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025 People’s Republic of China Seth L. Alper, Renal Division and Molecular and Vascular Medicine Unit, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USA Heng-Jing Cui, Department of Pharmacy, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025 People’s Republic of China Yang Lu, Department of Pharmacy, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025 People’s Republic of China Lin Zheng, Department of Pathology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025 People’s Republic of China Zhao-Wen Yan, Department of Pathology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025 People’s Republic of China Guo-Hui Fu, Department of Pathology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025 People’s Republic of China Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 6 Jan 2012 | 5:47 pm CET

RhoB is associated with the anti-angiogenic effects of celiac patient transglutaminase 2-targeted autoantibodies Abstract   Celiac patient-derived anti-transglutaminase 2 (TG2) antibodies disturb several steps in angiogenesis, but the detailed molecular basis is not known. Therefore, we here analyzed by microarray technology the expression of a set of genes related to angiogenesis and endothelial cell biology in order to identify factors that could explain our previous data related to vascular biology in the context of celiac disease. To this end, in vitro models using human umbilical vein endothelial cells (HUVECs) or in vivo models of angiogenesis were used. A total of 116 genes were analyzed after treatment with celiac patient autoantibodies against TG2. Compared to treatment with control IgA celiac patient, total IgA induced a consistent expression change of 10 genes, the up-regulation of four and down-regulation of six. Of these genes the up-regulated RhoB was selected for further studies. RhoB expression was found to be up-regulated at both messenger RNA and protein level in response to celiac patient total IgA as well as anti-TG2-specific antibody derived from a celiac patient. Interestingly, down-regulation of RhoB by specific small interfering RNA treatment in endothelial cells could rescue the deranged endothelial length and tubule formation caused by celiac disease autoantibodies. RhoB function is controlled by its post-translational modification by farnesylation. This modification of RhoB required for its correct function can be prevented by the cholesterol lowering drug simvastatin, which was also able to abolish the anti-angiogenic effects of celiac anti-TG2 autoantibodies. Taken together, our results would suggest that RhoB plays a key role in the response of endothelial cells to celiac disease-specific anti-TG2 autoantibodies. Content Type Journal Article Category Original Article Pages 1-10 DOI 10.1007/s00109-011-0853-0 Authors Stefania Martucciello, Pediatric Research Center, University of Tampere and Tampere University Hospital, Finn-Medi 3, School of Medicine, 33014 University of Tampere, Finland Miha Lavric, Department of Life Sciences, University of Trieste, Trieste, Italy Toth Boglarka, Department of Biochemistry and Molecular Biology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary Ilma Korponay-Szabo, Pediatric Research Center, University of Tampere and Tampere University Hospital, Finn-Medi 3, School of Medicine, 33014 University of Tampere, Finland Cristina Nadalutti, Pediatric Research Center, University of Tampere and Tampere University Hospital, Finn-Medi 3, School of Medicine, 33014 University of Tampere, Finland Essi Myrsky, Pediatric Research Center, University of Tampere and Tampere University Hospital, Finn-Medi 3, School of Medicine, 33014 University of Tampere, Finland Tiina Rauhavirta, Pediatric Research Center, University of Tampere and Tampere University Hospital, Finn-Medi 3, School of Medicine, 33014 University of Tampere, Finland Carla Esposito, Department of Chemistry, University of Salerno, Fisciano, Italy Ana-Marija Sulic, Pediatric Research Center, University of Tampere and Tampere University Hospital, Finn-Medi 3, School of Medicine, 33014 University of Tampere, Finland Daniele Sblattero, Department Medical Sciences and IRCAD, University of Eastern Piedmont, Novara, Italy Roberto Marzari, Department of Life Sciences, University of Trieste, Trieste, Italy Markku Mäki, Pediatric Research Center, University of Tampere and Tampere University Hospital, Finn-Medi 3, School of Medicine, 33014 University of Tampere, Finland Katri Kaukinen, School of Medicine, University of Tampere, Finn-Medi 3, School of Medicine, 33014 University of Tampere, Finland Katri Lindfors, Pediatric Research Center, University of Tampere and Tampere University Hospital, Finn-Medi 3, School of Medicine, 33014 University of Tampere, Finland Sergio Caja, Pediatric Research Center, University of Tampere and Tampere University Hospital, Finn-Medi 3, School of Medicine, 33014 University of Tampere, Finland Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 5 Jan 2012 | 6:07 pm CET

The mitogen-activated protein kinase (MAPK) cascade controls phosphatase and tensin homolog (PTEN) expression through multiple mechanisms Abstract   The mitogen-activated protein kinase (MAPK) and PI3K pathways are regulated by extensive crosstalk, occurring at different levels. In tumors, transactivation of the alternate pathway is a frequent “escape” mechanism, suggesting that combined inhibition of both pathways may achieve synergistic antitumor activity. Here we show that, in the M14 melanoma model, simultaneous inhibition of both MEK and mammalian target of rapamycin (mTOR) achieves synergistic effects at suboptimal concentrations, but becomes frankly antagonistic in the presence of relatively high concentrations of MEK inhibitors. This observation led to the identification of a novel crosstalk mechanism, by which either pharmacologic or genetic inhibition of constitutive MEK signaling restores phosphatase and tensin homolog (PTEN) expression, both in vitro and in vivo, and inhibits downstream signaling through AKT and mTOR, thus bypassing the need for double pathway blockade. This appears to be a general regulatory mechanism and is mediated by multiple mechanisms, such as MAPK-dependent c-Jun and miR-25 regulation. Finally, PTEN upregulation appears to be a major effector of MEK inhibitors’ antitumor activity, as cancer cells in which PTEN is inactivated are consistently more resistant to the growth inhibitory and anti-angiogenic effects of MEK blockade. Content Type Journal Article Category Original Article Pages 1-13 DOI 10.1007/s00109-011-0844-1 Authors Ludovica Ciuffreda, Division of Medical Oncology A, the Laboratory of Experimental Preclinical Chemotherapy and Translational Oncogenomics, Regina Elena National Cancer Institute, Chianesi, n. 53, 00144 Rome, Italy Cristina Di Sanza, Division of Medical Oncology A, the Laboratory of Experimental Preclinical Chemotherapy and Translational Oncogenomics, Regina Elena National Cancer Institute, Chianesi, n. 53, 00144 Rome, Italy Ursula Cesta Incani, Division of Medical Oncology A, the Laboratory of Experimental Preclinical Chemotherapy and Translational Oncogenomics, Regina Elena National Cancer Institute, Chianesi, n. 53, 00144 Rome, Italy Adriana Eramo, Department of Hematology, Oncology, and Molecular Medicine, Istituto Superiore di Sanità, Rome, 00161 Italy Marianna Desideri, Division of Medical Oncology A, the Laboratory of Experimental Preclinical Chemotherapy and Translational Oncogenomics, Regina Elena National Cancer Institute, Chianesi, n. 53, 00144 Rome, Italy Francesca Biagioni, Division of Medical Oncology A, the Laboratory of Experimental Preclinical Chemotherapy and Translational Oncogenomics, Regina Elena National Cancer Institute, Chianesi, n. 53, 00144 Rome, Italy Daniela Passeri, Anatomic Pathology, Tor Vergata University of Rome, Rome, 00173 Italy Italia Falcone, Division of Medical Oncology A, the Laboratory of Experimental Preclinical Chemotherapy and Translational Oncogenomics, Regina Elena National Cancer Institute, Chianesi, n. 53, 00144 Rome, Italy Giovanni Sette, Department of Hematology, Oncology, and Molecular Medicine, Istituto Superiore di Sanità, Rome, 00161 Italy Paola Bergamo, Division of Medical Oncology A, the Laboratory of Experimental Preclinical Chemotherapy and Translational Oncogenomics, Regina Elena National Cancer Institute, Chianesi, n. 53, 00144 Rome, Italy Andrea Anichini, Human Tumor Immunobiology Unit, Fondazione IRCCS Istituto Nazionale per lo Studio e la Cura dei Tumori, Milan, 20133 Italy Kanaga Sabapathy, Laboratory of Molecular Carcinogenesis, National Cancer Centre, Singapore, 169610 Singapore James A. McCubrey, Department of Microbiology and Immunology, Brody School of Medicine, East Carolina University, Greenville, NC 27834, USA Maria Rosaria Ricciardi, Department of Cellular Biotechnologies and Hematology, University of Rome ‘La Sapienza’, Rome, 00161 Italy Agostino Tafuri, Department of Cellular Biotechnologies and Hematology, University of Rome ‘La Sapienza’, Rome, 00161 Italy Giovanni Blandino, Division of Medical Oncology A, the Laboratory of Experimental Preclinical Chemotherapy and Translational Oncogenomics, Regina Elena National Cancer Institute, Chianesi, n. 53, 00144 Rome, Italy Augusto Orlandi, Anatomic Pathology, Tor Vergata University of Rome, Rome, 00173 Italy Ruggero De Maria, Department of Hematology, Oncology, and Molecular Medicine, Istituto Superiore di Sanità, Rome, 00161 Italy Francesco Cognetti, Division of Medical Oncology A, the Laboratory of Experimental Preclinical Chemotherapy and Translational Oncogenomics, Regina Elena National Cancer Institute, Chianesi, n. 53, 00144 Rome, Italy Donatella Del Bufalo, Division of Medical Oncology A, the Laboratory of Experimental Preclinical Chemotherapy and Translational Oncogenomics, Regina Elena National Cancer Institute, Chianesi, n. 53, 00144 Rome, Italy Michele Milella, Division of Medical Oncology A, the Laboratory of Experimental Preclinical Chemotherapy and Translational Oncogenomics, Regina Elena National Cancer Institute, Chianesi, n. 53, 00144 Rome, Italy Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 4 Jan 2012 | 7:58 am CET

Sox9/Sox6 and Sp1 are involved in the insulin-like growth factor-I-mediated upregulation of human type II collagen gene expression in articular chondrocytes Abstract   Type II collagen is a marker of articular cartilage encoded by the COL2A1 gene. The nature of the trans factors involved in the upregulation of this gene by insulin-like growth factor-I (IGF-I) remains unclear. We found that IGF-I increased type II collagen synthesis by a transcriptional control mechanism involving a 715-bp region within the COL2A1 first-intron specific enhancer. The overproduction of L-Sox5/Sox6/Sox9 and Sp1 and decoy experiments targeting these factors demonstrated their action in concert in IGF-I trans-activation. These results were supported by the data obtained in knockdown experiments in which siRNA against Sox9/Sox6 and Sp1 prevented the IGF-I-induced increase in collagen II production. Indeed, each of these trans-activators increased the expression of others. IGF-I increased the binding of Sox9 and Sp1/Sp3 to their cis elements in the enhancer, and we provide the first evidence of Sox9 interaction with the promoter by chromatin immunoprecipitation. Interactions with COL2A1 were also observed for Sp1, p300/CBP, and Tip60. Finally, a physical interaction between Sox9, p300, Sp3, and Sp1 was detected. These data demonstrate the role of Sox9, Sp1/Sp3, and euchromatin-associated factors (p300, Tip60) in the IGF-I-induced upregulation of COL2A1, indicating possible use of this growth factor in articular cartilage engineering applications to promote repair in patients with degenerative diseases, such as osteoarthritis. Content Type Journal Article Category Original Article Pages 1-18 DOI 10.1007/s00109-011-0842-3 Authors Emmanuelle Renard, Laboratoire Matrice Extracellulaire et Pathologie, IFR ICORE 146, Université de Caen/Basse-Normandie, UFR de Médecine, CHU niveau 3, Avenue de la Côte de Nacre, 14032 Caen Cedex, France Benoît Porée, Laboratoire Matrice Extracellulaire et Pathologie, IFR ICORE 146, Université de Caen/Basse-Normandie, UFR de Médecine, CHU niveau 3, Avenue de la Côte de Nacre, 14032 Caen Cedex, France Christos Chadjichristos, Laboratoire Matrice Extracellulaire et Pathologie, IFR ICORE 146, Université de Caen/Basse-Normandie, UFR de Médecine, CHU niveau 3, Avenue de la Côte de Nacre, 14032 Caen Cedex, France Magdalini Kypriotou, Laboratoire Matrice Extracellulaire et Pathologie, IFR ICORE 146, Université de Caen/Basse-Normandie, UFR de Médecine, CHU niveau 3, Avenue de la Côte de Nacre, 14032 Caen Cedex, France Laure Maneix, Laboratoire Matrice Extracellulaire et Pathologie, IFR ICORE 146, Université de Caen/Basse-Normandie, UFR de Médecine, CHU niveau 3, Avenue de la Côte de Nacre, 14032 Caen Cedex, France Nicolas Bigot, Laboratoire Matrice Extracellulaire et Pathologie, IFR ICORE 146, Université de Caen/Basse-Normandie, UFR de Médecine, CHU niveau 3, Avenue de la Côte de Nacre, 14032 Caen Cedex, France Florence Legendre, Laboratoire Matrice Extracellulaire et Pathologie, IFR ICORE 146, Université de Caen/Basse-Normandie, UFR de Médecine, CHU niveau 3, Avenue de la Côte de Nacre, 14032 Caen Cedex, France David Ollitrault, Laboratoire Matrice Extracellulaire et Pathologie, IFR ICORE 146, Université de Caen/Basse-Normandie, UFR de Médecine, CHU niveau 3, Avenue de la Côte de Nacre, 14032 Caen Cedex, France Benoît De Crombrugghe, Department of Molecular Genetics, University of Texas, M.D. Anderson Cancer Center, Holcombe Boulevard, Houston, TX 77030, USA Frédéric Malléin-Gérin, Laboratoire de Biologie et Ingénierie du Cartilage, Institut de Biologie et Chimie des Protéines, UMR 5086 CNRS/Université Claude Bernard Lyon 1-IFR 128 Biosciences Lyon-Gerland, 7 Passage du Vercors, 69367 Lyon Cedex 07, France Safa Moslemi, Laboratoire Matrice Extracellulaire et Pathologie, IFR ICORE 146, Université de Caen/Basse-Normandie, UFR de Médecine, CHU niveau 3, Avenue de la Côte de Nacre, 14032 Caen Cedex, France Magali Demoor, Laboratoire Matrice Extracellulaire et Pathologie, IFR ICORE 146, Université de Caen/Basse-Normandie, UFR de Médecine, CHU niveau 3, Avenue de la Côte de Nacre, 14032 Caen Cedex, France Karim Boumediene, Laboratoire Matrice Extracellulaire et Pathologie, IFR ICORE 146, Université de Caen/Basse-Normandie, UFR de Médecine, CHU niveau 3, Avenue de la Côte de Nacre, 14032 Caen Cedex, France Philippe Galéra, Laboratoire Matrice Extracellulaire et Pathologie, IFR ICORE 146, Université de Caen/Basse-Normandie, UFR de Médecine, CHU niveau 3, Avenue de la Côte de Nacre, 14032 Caen Cedex, France Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 4 Jan 2012 | 7:58 am CET

Identification of F-box only protein 7 as a negative regulator of NF-kappaB signaling Abstract The Nuclear factor κB (NF-κB) signaling pathway controls important cellular events such as cell proliferation, differentiation, apoptosis and immune responses. Pathway activation occurs rapidly upon TNFα stimulation and is highly dependent on ubiquitination events. Using cytoplasmic to nuclear translocation of the NF-κB transcription factor family member p65 as a read-out, we screened a synthetic siRNA library targeting enzymes involved in ubiquitin conjugation and de-conjugation for modifiers of regulatory ubiquitination events in NF-κB signaling. We identified F-Box protein only 7 (FBXO7), a component of SCF-ubiquitin ligase complexes, as a negative regulator of NF-κB signaling. FBXO7 binds to, and mediates ubiquitin conjugation to cIAP1 and TRAF2, resulting in decreased RIP1 ubiquitination and lowered NF-κB signaling activity. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 2 Jan 2012 | 3:36 pm CET

Large Animal Induced Pluripotent Stem Cells as Pre-Clinical Models For Studying Human Disease Abstract The derivation of human embryonic stem cells (ESCs) and subsequently human induced pluripotent stem cells (iPSCs) has energized regenerative medicine research and enabled seemingly limitless applications. While small animal models, such as mouse models, have played an important role in the progression of the field, typically, they are poor representations of the human disease phenotype. As an alternative, large animal models should be explored as a potentially better approach for clinical translation of cellular therapies. However, only fragmented information regarding the derivation, characterization, and clinical usefulness of pluripotent large animal cells is currently available. Here we briefly review the latest advances regarding the derivation and use of large animal iPSCs. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 2 Jan 2012 | 3:36 pm CET

Mitochondrial connexin 43 impacts on respiratory complex I activity and mitochondrial oxygen consumption Abstract Connexin 43 (Cx43) is present at the sarcolemma and the inner membrane of cardiomyocyte subsarcolemmal mitochondria (SSM). Lack or inhibition of mitochondrial Cx43 is associated with reduced mitochondrial potassium influx, which might affect mitochondrial respiration. Therefore, we analyzed the importance of mitochondrial Cx43 for oxygen consumption. Acute inhibition of Cx43 in rat left ventricular (LV) SSM by 18α glycyrrhetinic acid (GA) or Cx43 mimetic peptides (Cx43-MP) reduced ADP-stimulated complex I respiration and ATP generation. Chronic reduction of Cx43 in conditional knockout mice (Cx43Cre-ER(T)/fl + 4-OHT, 5-10% of Cx43 protein compared to control Cx43fl/fl mitochondria) reduced ADP-stimulated complex I respiration of LV SSM to 47.8±2.4 nmol O2/min*mg protein (n=8) from 61.9±7.4 nmol O2/min*mg protein in Cx43fl/fl mitochondria (n=10, p<0.05), while complex II respiration remained unchanged. The LV complex I activities (% of citrate synthase activity) of Cx43Cre-ER(T)/fl+4-OHT mice (16.1±0.9%, n=9) were lower than in Cx43fl/fl mice (19.8±1.3%, n=8, p<0.05); complex II activities were similar between genotypes. Supporting the importance of Cx43 for respiration, in Cx43-overexpressing HL-1 cardiomyocytes complex I respiration was increased, whereas complex II respiration was unaffected. Taken together, mitochondrial Cx43 is required for optimal complex I activity and respiration and thus mitochondrial ATP-production. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 30 Dec 2011 | 5:04 pm CET

Small molecule regulators of postnatal Nkx2.5 cardiomyoblast proliferation and differentiation Abstract While recent data have supported the capacity for a neonatal heart to undergo cardiomyogenesis, it is unclear whether these new cardiomyocytes arise from an immature cardiomyoblast population or from the division of mature cardiomyocytes. By following the expression eGFP in an Nkx2.5 enhancer-eGFP transgenic mice, we have identified a population of immature cells that can undergo cardiomyogenic as well as smooth muscle cell differentiation in the neonatal heart. Here, we examined growth factors and small molecule regulators that potentially regulate the proliferation and cardiomyogenic vs smooth muscle cell differentiation of neonatal Nkx2.5-GFP+ cells in vitro. We found that A83-01 (A83), an inhibitor of TGF-βRI, was able to induce an expansion of neonatal Nkx2.5-eGFP+ cells. In addition, the ability of A83 to expand eGFP+ cells in culture was dependent on signaling from MEK since treatment with a MEK inhibitor, PD0325901, abolished this effect. On the other hand, activation of neonatal Nkx2.5-eGFP+ cells with TGF-β1, but not Activin A nor BMP2, led to smooth muscle cell differentiation, an effect that can be reversed by treatment with A83. In summary, small molecule inhibition of TGFβ signaling may be a promising strategy to induce the expansion of a rare population of postnatal cardiomyoblasts. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 30 Dec 2011 | 5:04 pm CET

Novel phosphatidylinositol 3-kinase inhibitor NVP-BKM120 induces apoptosis in myeloma cells and shows synergistic anti-myeloma activity with dexamethasone Abstract   NVP-BKM120 is a novel phosphatidylinositol 3-kinase (PI3K) inhibitor and is currently being investigated in phase I clinical trials in solid tumors. This study aimed to evaluate the therapeutic efficacy of BKM120 in multiple myeloma (MM). BKM120 induces cell growth inhibition and apoptosis in both MM cell lines and freshly isolated primary MM cells. However, BKM120 only shows limited cytotoxicity toward normal lymphocytes. The presence of MM bone marrow stromal cells, insulin-like growth factor, or interleukin-6 does not affect BKM120-induced tumor cell apoptosis. More importantly, BKM120 treatment significantly inhibits tumor growth in vivo and prolongs the survival of myeloma-bearing mice. In addition, BKM120 shows synergistic cytotoxicity with dexamethasone in dexamethasone-sensitive MM cells. Low doses of BKM120 and dexamethasone, each of which alone has limited cytotoxicity, induce significant cell apoptosis in MM.1S and ARP-1. Mechanistic study shows that BKM120 exposure causes cell cycle arrest by upregulating p27 (Kip1) and downregulating cyclin D1 and induces caspase-dependent apoptosis by downregulating antiapoptotic XIAP and upregulating expression of cytotoxic small isoform of Bim, BimS. In summary, our findings demonstrate the in vitro and in vivo anti-MM activity of BKM120 and suggest that BKM120 alone or together with other MM chemotherapeutics, particularly dexamethasone, may be a promising treatment for MM. Content Type Journal Article Category Original Article Pages 1-12 DOI 10.1007/s00109-011-0849-9 Authors Yuhuan Zheng, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Jing Yang, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Jianfei Qian, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Liang Zhang, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Yong Lu, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Haiyan Li, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Heather Lin, Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA Yongsheng Lan, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Zhiqiang Liu, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Jin He, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Sungyoul Hong, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Sheeba Thomas, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Jatin Shah, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Veera Baladandayuthapani, Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA Larry W. Kwak, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Qing Yi, Department of Lymphoma/Myeloma, Division of Cancer Medicine, Center for Cancer Immunology Research, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Unit 0903, Houston, TX 77030, USA Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 30 Dec 2011 | 8:09 am CET

Cystamine attenuates lupus-associated apoptosis of ventricular tissue by suppressing both intrinsic and extrinsic pathway Abstract Cystamine, a disulphide metabolite, has been demonstrated to ameliorate various lupus-associated tissue damages by animal models. However, effects of cystamine on apoptosis of cardiac tissue, a main cardiac damage attributing to lupus, are not clear. Therefore, we aimed to investigate whether cystamine possesses anti-apoptotic effects with emphasis on left ventricular (LV) tissue of lupus-prone mice NZB/W-F1. Cystamine treatment was performed by daily intraperitoneal administration. Morphology and apoptotic status of ventricular tissues in the treated mice were assessed by microscopy and TUNEL assay respectively. Levels of apoptotic biomarkers were determined using immunoblot. Our results revealed that cystamine significantly attenuated the apoptosis of LV tissues in NZB/W-F1 mice, whereas the morphology of the tissues was slightly altered. Additionally, cystamine reduced level of Fas and inhibited activation of caspase-8. Cystamine also increased level of Bcl-2 and phosphorylation of Bad, and decreased level of Bad and truncated Bid (tBid). Moreover, level of cytosolic cytochrome c and Apaf-1, and activation of caspase-9 and -3 were suppressed in response to cystamine treatment. In Balb/c mice, as normal control mice, changes in cell morphology and levels of the tested apoptotic components were found insignificant in the LV tissues. These findings indicate that cystamine treatment attenuates apoptosis of LV tissues of NZB/W-F1 mice through suppressing both intrinsic and extrinsic apoptotic pathways. Therefore, cystamine is considered beneficial to alleviating lupus-associated cardiac damages. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 28 Dec 2011 | 9:42 pm CET

Antimyeloma activity of the sesquiterpene lactone cnicin: impact on Pim-2 kinase as a novel therapeutic target Abstract   Despite recent advances in therapy, multiple myeloma, the second most common hematologic tumor in the Western world, is still incurable. Identification of substances that display a wide range of tumor-killing activities and target cancer-specific pathways constitute a basis for the development of novel therapies. In this study, we investigate the cytotoxic effect of the natural substance cnicin in multiple myeloma. Cnicin treatment reveals potent antiproliferative effects and induces cell death in cell lines and primary myeloma cells even in the presence of survival cytokines and the tumor microenvironment. Other cell lines of hematopoietic origin also succumb to cell death whereas stromal cells and endothelial cells are unaffected. We show that activation of caspases, accumulation of reactive oxygen species and downregulation of nuclear factor kappa-light-chain-enhancer of activated B cell contribute to the cytotoxic effects of cnicin. Microarray analysis reveals downregulation of Pim-2, a serine/threonine kinase. We provide evidence that Pim-2 constitutes a new survival kinase for myeloma cells in vitro and is highly expressed in malignant but not in normal plasma cells in vivo. Combining cnicin with current standard or experimental therapeutics leads to enhanced cell death. Thus, our data indicate that cnicin induces myeloma cell death via several pathways and reveals Pim-2 as a novel target. These findings provide a rational for further evaluation of cnicin as a new anti-tumor drug and underline the potential of sesquiterpene lactones in tumor therapy. Content Type Journal Article Category Original Article Pages 1-13 DOI 10.1007/s00109-011-0848-x Authors Karin Jöhrer, Tyrolean Cancer Research Institute, Innrain 66, 6020 Innsbruck, Austria Marlene Obkircher, Tyrolean Cancer Research Institute, Innrain 66, 6020 Innsbruck, Austria Daniel Neureiter, Institute of Pathology, University Hospital Salzburg, Müllner Hauptstrasse 48, 5020 Salzburg, Austria Johanna Parteli, Tyrolean Cancer Research Institute, Innrain 66, 6020 Innsbruck, Austria Claudia Zelle-Rieser, Tyrolean Cancer Research Institute, Innrain 66, 6020 Innsbruck, Austria Eva Maizner, Tyrolean Cancer Research Institute, Innrain 66, 6020 Innsbruck, Austria Johann Kern, Department of Internal Medicine V, Innsbruck Medical University, Innsbruck, Austria Martin Hermann, KMT Laboratory, Department of Visceral-, Transplant- and Thoracic Surgery, Innsbruck Medical University, Innsbruck, Austria Frank Hamacher, Laboratory for Immunological and Molecular Cancer Research, Third Medical Department, University Hospital Salzburg, Salzburg, Austria Olaf Merkel, Laboratory for Immunological and Molecular Cancer Research, Third Medical Department, University Hospital Salzburg, Salzburg, Austria Nathalie Wacht, Laboratory for Immunological and Molecular Cancer Research, Third Medical Department, University Hospital Salzburg, Salzburg, Austria Christian Zidorn, Institute of Pharmacy, Department of Pharmacognosy, University of Innsbruck, Innsbruck, Austria Marcel Scheideler, Institute for Genomics and Bioinformatics, Graz University of Technology, Petersgasse 14, 8010 Graz, Austria Richard Greil, Tyrolean Cancer Research Institute, Innrain 66, 6020 Innsbruck, Austria Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 28 Dec 2011 | 5:45 pm CET

Distinct regulation of nNOS and iNOS by CB2 receptor in remote delayed neurodegeneration Abstract   Hemicerebellectomy results in remote delayed degeneration of precerebellar neurons. We have reported that such a lesion induces type 2 cannabinoid receptor (CB2) expression in precerebellar neurons and that stimulation of CB2, but not CB1, has neuroprotective effects. In this study, we found that in the same model, the CB2 agonist JWH-015 enhances neuronal nitric oxide synthase (nNOS) expression in axotomized neurons and that CB2-mediated neuroprotection is abrogated by pharmacological inhibition of nNOS. JWH-015 prevented the axotomy-induced upregulation of inducible NOS (iNOS) in astrocytes but had no effect on endothelial NOS (eNOS). In addition, we observed that JWH-015 significantly reduces hemicerebellectomy-induced neuroinflammatory responses and oxidative/nitrative stress. With regard to the signaling pathways of CB2/nNOS-mediated neuroprotection, we noted nNOS-dependent modulation of the expression of anti-oxidative (Hsp70) and anti-apoptotic (Bcl-2) proteins. These findings shed light on the interactions between the endocannabinoid and nitrergic systems after focal brain injury, implicating distinct functions of nNOS activation and iNOS inhibition in CB2 signaling, which protect neurons from axotomy-induced cell death. Content Type Journal Article Category Original Article Pages 1-17 DOI 10.1007/s00109-011-0846-z Authors S. Oddi, Department of Biomedical Sciences, University of Teramo, Piazza Aldo Moro 45, 64100 Teramo, Italy L. Latini, Santa Lucia Foundation I.R.C.C.S., Via del Fosso di Fiorano 65, 00143 Rome, Italy M. T. Viscomi, Santa Lucia Foundation I.R.C.C.S., Via del Fosso di Fiorano 65, 00143 Rome, Italy E. Bisicchia, Department of Biomedical Sciences, University of Teramo, Piazza Aldo Moro 45, 64100 Teramo, Italy M. Molinari, Santa Lucia Foundation I.R.C.C.S., Via del Fosso di Fiorano 65, 00143 Rome, Italy M. Maccarrone, Department of Biomedical Sciences, University of Teramo, Piazza Aldo Moro 45, 64100 Teramo, Italy Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 23 Dec 2011 | 5:55 pm CET

PDGFR-β positive telocytes in skeletal muscle interstitium Abstract Telocytes (TCs) represent a new cell type recently described in mammalian skeletal muscle interstitium as well as in other organs. TCs have a specific morphology and phenotype, both in situ and in vitro. TCs are cells with long and slender cell prolongations, in contact with other interstitial cells, nerve fibers, blood capillaries and resident stem cells in niches. Our aim was to investigate the potential contribution of TCs to micro-vascular networks by immunofluorescent labelling of specific angiogenic growth factors and receptors. We found that in human skeletal muscle TCs were constantly located around intermediate and small blood vessels and endomysial capillaries. Epi-fluorescence and laser confocal microscopy showed that TCs express c-kit, PDGFRβ and VEGF, both in situ and in vitro. TCs were constantly located in the perivascular or pericapillary space, as confirmed by double staining of c-kit/CD31, PDGFRβ/CD31 and PDGFRβ/αSMA, respectively. Electron microscopy (EM) differentiated between pericytes and other cell types. Laminin labelling showed that TCs are not enclosed or surrounded by a basal lamina in contrast to mural cells. In conclusion: a) PDGFRβ could be used as a marker for TCs and b) TCs are presumably a transitional population in the complex process of mural cell recruitment during angiogenesis and vascular remodelling. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 21 Dec 2011 | 11:52 am CET

Regenerative potential of glycosaminoglycans for skin and bone Abstract   To meet the growing need for tissue replacement materials for our aging population, the development of new adaptive biomaterials is essential. The tissues with the highest demand for implant materials are skin and bone. These tissues share various similarities, including signaling pathways and extracellular matrix composition. Glycosaminoglycans such as hyaluronan and chondroitin sulfate are the major organic extracellular matrix components. They modulate the attraction of skin and bone precursor cells and their subsequent differentiation and gene expression and regulate the action of proteins essential to bone and skin regeneration. The precise action of glycosaminoglycans varies according to their structural composition mainly in respect to the degree of sulfation and polymer length. Changes in the glycosaminoglycan composition are frequently seen in physiological and pathological remodeling processes, such as bone formation or scaring. Here, we review the current state of knowledge of how the most common glycosaminoglycan, chondroitin sulfate and hyaluronan, interact with bone and skin cells, and summarize their potential in tissue engineering for skeletal and skin diseases. Content Type Journal Article Category Review Pages 1-11 DOI 10.1007/s00109-011-0843-2 Authors Juliane Salbach, Division of Endocrinology, Diabetes, and Metabolic Bone Diseases, Department of Medicine III, Dresden Technical University Medical Center, Fetscherstraße 74, 01307 Dresden, Germany Tilman D. Rachner, Division of Endocrinology, Diabetes, and Metabolic Bone Diseases, Department of Medicine III, Dresden Technical University Medical Center, Fetscherstraße 74, 01307 Dresden, Germany Martina Rauner, Division of Endocrinology, Diabetes, and Metabolic Bone Diseases, Department of Medicine III, Dresden Technical University Medical Center, Fetscherstraße 74, 01307 Dresden, Germany Ute Hempel, Institute of Physiological Chemistry, Technical University Dresden, Dresden, Germany Ulf Anderegg, Department of Dermatology, Venerology and Allergology, University of Leipzig, Leipzig, Germany Sandra Franz, Department of Dermatology, Venerology and Allergology, University of Leipzig, Leipzig, Germany Jan-Christoph Simon, Department of Dermatology, Venerology and Allergology, University of Leipzig, Leipzig, Germany Lorenz C. Hofbauer, Division of Endocrinology, Diabetes, and Metabolic Bone Diseases, Department of Medicine III, Dresden Technical University Medical Center, Fetscherstraße 74, 01307 Dresden, Germany Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 20 Dec 2011 | 5:46 pm CET

Interaction between pathogenic proteins in neurodegenerative disorders Abstract The misfolding and progressive aggregation of specific proteins in selective regions of the nervous system is a seminal occurrence in many neurodegenerative disorders, and the interaction between pathological/toxic proteins to cause neurodegeneration is a hot topic of current neuroscience research. Despite clinical, genetic, and experimental differences, increasing evidence indicates considerable overlap between synucleinopathies, tauopathies and other protein-misfolding diseases. Inclusions, often characteristic hallmarks of these disorders, suggest interactions of pathological proteins enganging common downstream pathways. Novel findings that have shifted our understanding in the role of pathologic proteins in the pathogenesis of Alzheimer, Parkinson, Huntington, and prion diseases, have confirmed correlations/overlaps between these and other neurodegenerative disorders. Emerging evidence, in addition to synergistic effects of tau protein, amyloid β, α-synuclein, and other pathologic proteins, suggests that prion-like induction and spreading, involving secreted proteins, are major pathogenic mechanisms in various neurodegenerative diseases, depending on genetic backgrounds and environmental factors. The elucidation of the basic molecular mechanisms underlying the interaction and spreading of pathogenic proteins, suggesting a dualism or triad of neurodegeneration in protein-misfolding disorders, is a major challenge for modern neuroscience, in order to provide a deeper insight into their pathogenesis as a basis of effective diagnosis and treatment. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 16 Dec 2011 | 6:06 pm CET

Sequential targeting of CFTR by BAC vectors generates a novel pig model of cystic fibrosis Abstract   Cystic fibrosis (CF) is the most common lethal inherited disease in Caucasians and is caused by mutations in the CFTR gene. The disease is incurable and medical treatment is limited to the amelioration of symptoms or secondary complications. A comprehensive understanding of the disease mechanisms and the development of novel treatment options require appropriate animal models. Existing CF mouse models fail to reflect important aspects of human CF. We thus generated a CF pig model by inactivating the CFTR gene in primary porcine cells by sequential targeting using modified bacterial artificial chromosome vectors. These cells were then used to generate homozygous CFTR mutant piglets by somatic cell nuclear transfer. The homozygous CFTR mutants lack CFTR protein expression and display severe malformations in the intestine, respiratory tract, pancreas, liver, gallbladder, and male reproductive tract. These phenotypic abnormalities closely resemble both the human CF pathology as well as alterations observed in a recently published CF pig model which was generated by a different gene targeting strategy. Our new CF pig model underlines the value of the CFTR-deficient pig for gaining new insight into the disease mechanisms of CF and for the development and evaluation of new therapeutic strategies. This model will furthermore increase the availability of CF pigs to the scientific community. Content Type Journal Article Category Original Article Pages 1-12 DOI 10.1007/s00109-011-0839-y Authors N. Klymiuk, Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Straße 25, 81377 Munich, Germany L. Mundhenk, Department of Veterinary Pathology, Faculty of Veterinary Medicine, Freie Universität Berlin, Robert-von-Ostertag-Straße 15, 14163 Berlin, Germany K. Kraehe, Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Straße 25, 81377 Munich, Germany A. Wuensch, Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Straße 25, 81377 Munich, Germany S. Plog, Department of Veterinary Pathology, Faculty of Veterinary Medicine, Freie Universität Berlin, Robert-von-Ostertag-Straße 15, 14163 Berlin, Germany D. Emrich, Institute of Veterinary Pathology, Ludwig-Maximilians-Universität München, Veterinärstraße 13, 80539 Munich, Germany M. C. Langenmayer, Institute of Veterinary Pathology, Ludwig-Maximilians-Universität München, Veterinärstraße 13, 80539 Munich, Germany M. Stehr, Department of Pediatric Surgery, Dr. von Hauner Children’s Hospital, Ludwig-Maximilians-Universität München, Lindwurmstraße 4, 80337 Munich, Germany A. Holzinger, Division of Neonatology, Dr. von Hauner Children’s Hospital, Ludwig-Maximilians-Universität München, Lindwurmstraße 4, 80337 Munich, Germany C. Kröner, Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Straße 25, 81377 Munich, Germany A. Richter, Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Straße 25, 81377 Munich, Germany B. Kessler, Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Straße 25, 81377 Munich, Germany M. Kurome, Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Straße 25, 81377 Munich, Germany M. Eddicks, Clinic for Swine, Ludwig-Maximilians-Universität München, Sonnenstraße 16, 85764 Oberschleißheim, Germany H. Nagashima, Laboratory of Developmental Engineering, Department of Life Science, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama, Kawasaki, 214-8571 Japan K. Heinritzi, Clinic for Swine, Ludwig-Maximilians-Universität München, Sonnenstraße 16, 85764 Oberschleißheim, Germany A. D. Gruber, Department of Veterinary Pathology, Faculty of Veterinary Medicine, Freie Universität Berlin, Robert-von-Ostertag-Straße 15, 14163 Berlin, Germany E. Wolf, Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Straße 25, 81377 Munich, Germany Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 14 Dec 2011 | 5:41 pm CET

Intermittent Hypobaric Hypoxia applicability in myocardial infarction prevention and recovery Abstract Intermittent Hypobaric Hypoxia (IHH) has been the focus of important research in cardioprotection, and it has been associated with several mechanisms. IHH inhibits prolyl hydroxylases (PHD) activity, increasing the stabilization of hypoxia inducible factor-1 (HIF-1) and activating crucial adaptative genes. It has been hence suggested that IHH might be a simple intervention, which may offer a thoughtful benefits to patients with acute myocardial infarction and no complications. Nevertheless, several doubts exist as to whether IHH is a really safe technique, with little to no complications in post-myocardial infarction patients. IHH might produce instead unfavorable changes such as impairment of vascular hemodynamics and hypertensive response, increased risk of hemoconcentration and thrombosis, cardiac rhythm perturbations, coronary artery disease and heart failure, insulin resistance, steatohepatitis, and even high altitude pulmonary edema in susceptible or nonacclimatized subjects. Although intermittent and chronic exposures seem effective in cardioprotection, IHH safety issues have been mostly overlooked, so that assorted concerns should be raised about the opportunity to use IHH in the post-myocardial infarction period. Several IHH protocols used in some studies were also aggressive, which would hamper their widespread introduction within the clinical practice. As such, further research is needed before IHH can be widely advocated in myocardial infarction prevention and recovery. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 12 Dec 2011 | 3:38 pm CET

Immunogenicity of allogeneic mesenchymal stem cells Abstract Mesenchymal stem cells (MSCs) inhibit proliferation of allogeneic T cells and express low levels of major histocompatibility complex class I (MHCI), MHCII, and vascular adhesion molecule-1 (VCAM-1). We investigated whether their immunosuppressive properties and low immunophenotype protect allogeneic rat MSCs against cytotoxic lysis in vitro and result in a reduced immune response in vivo. Rat MSCs were partially protected against alloantigen-specific cytotoxic T cells in vitro. However, after treatment with IFN-γ and IL-1β, MSCs upregulated MHCI, MHCII, and VCAM-1, and cytotoxic lysis was significantly increased. In vivo, allogeneic T cells but not allogeneic MSCs induced upregulation of the activation markers CD25 and CD71 as well as downregulation of CD62L on CD4+ T cells from recipient rats. However, intravenous injection of allo-MSCs in rats led to the formation of alloantibodies with the capacity to facilitate complement-mediated lysis, although IgM levels were markedly decreased compared to animals that received T cells. The allo-MSC induced immune response was sufficient to lead to significantly reduced survival of subsequently injected allo-MSCs. Interestingly, no increased immunogenicity of IFN-γ stimulated allo-MSCs was observed in vivo. Both the loss of protection against cytotoxic lysis under inflammatory conditions and the induction of complement-activating antibodies will likely impact the utility of allogeneic MSCs for therapeutic applications. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 12 Dec 2011 | 3:37 pm CET

PDGFRα cells in mouse urinary bladder: A new class of interstitial cells Abstract Specific classes of interstitial cells exist in visceral organs and have been implicated in several physiological functions including pacemaking and mediators in neurotransmission. In the bladder, Kit+ interstitial cells have been reported to exist and have been suggested to be neuromodulators. More recently a second interstitial cell, that is identified using antibodies against platelet derived growth factor receptor-α (PDGFRα) has been described in the GI tract and has been implicated in enteric motor neurotransmission. In this study, we examined the distribution of PDGFRα+ cells in the murine urinary bladder and the relation that these cells may have with nerve fibers and smooth muscle cells. PDGFRα+ cells had a spindle shaped or stellate morphology and often possessed multiple processes that contacted one another forming a loose network. These cells were distributed throughout the bladder wall, being present in the lamina propria as well as throughout the muscularis of the detrusor. PDGFRα+ cells surrounded and were located between smooth muscle bundles and often came into close morphological association with intramural nerve fibers. These data describe a new class of interstitial cell that express a specific receptor within the bladder wall and provide morphological evidence for a possible neuromodulatory role in bladder function. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 12 Dec 2011 | 10:40 am CET

Identification of telocytes in the upper lamina propria of the human urinary tract Abstract The upper lamina propria (ULP) area of interstitial cells (IC) has been studied extensively in bladder, but is rather unexplored in the rest of the urinary tract. This cell layer is intriguing because of the localization directly underneath the urothelium, the intercellular contacts and the close relation with nerve endings and capillaries. In the present study we examine the ULP layer of IC in human renal pelvis, ureter and urethra, and we make a comparison with ULP IC in bladder. Tissue was obtained from normal areas in nephrectomy-, cystectomy- and prostatectomy-specimens, and processed for morphology, immunohistochemistry and electron microscopy. A morphological and immunohistochemical phenotype for the ULP IC was assessed and region-dependent differences were looked for. The ULP IC in renal pelvis, ureter and urethra had a similar ultrastructural phenotype, which differed somehow from that of bladder IC i.e.: thinner and longer cytoplasmic processes, no peripheral actin filaments, and presence of dense core granules and microtubules. Together with their immunohistochemical profile, these features are most compatible with the phenotype of telocytes, a recently discovered group of stromal cells. Based on their global ultrastructural and immunohistochemical phenotype ULP IC in human bladder should also be classified as telocytes. The most striking immunohistochemical finding was the variable expression of estrogen receptor (ER) and progesterone receptor (PR). The functional relevance of ULP telocytes in the urinary tract remains to be elucidated, and ER and PR might therefore be promising pharmacological research targets. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 7 Dec 2011 | 5:42 pm CET

Cross-talk between DNA methylation and active histone modifications regulates aberrant expression of ZAP70 in CLL Abstract Zeta-associated protein of 70 kDa (ZAP70) is a recognised adverse prognostic marker in CLL associated with enhanced B cell receptor signalling, significantly more aggressive disease course and poor overall survival. ZAP70 is ordinarily expressed in T cells where it has a crucial role in T cell receptor signalling, whereas its aberrant expression in CLL leads to enhanced B cell receptor signalling and significantly more aggressive disease course. Although much is known about the activation of ZAP70 following engagement of T cell receptor, there is little data on the regulation of ZAP70 gene expression in normal T cells or CLL. To understand the molecular events underpinning epigenetic regulation of ZAP70 gene in CLL, we have defined ZAP70 promoter region and outlined the regions crucial in regulating the gene activity. Following a direct comparison of ZAP70 positive and negative primary CLLs, we show ZAP70 promoter in expressing CLLs to be associated with a spectrum of active histone modifications, some of which are tightly linked to aberrant DNA methylation in CLL. Cross-talk between histone modifications and reduced DNA methylation culminates in transcriptional de-repression of ZAP70. Moreover, treatment with HDAC and DNA methylation inhibitors results in recovery of ZAP70 expression which provides a possible explanation for the failure of HDAC inhibitors in CLL treatment and might serve as a cautionary warning for their future use in treatment of this leukaemia. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd Quelle: Journal of Cellular and Molecular Medicine | 7 Dec 2011 | 4:28 pm CET

Circulating microRNAs: novel biomarkers for cardiovascular diseases Abstract   MicroRNAs (miRNAs) are a novel class of small, non-coding, single-stranded RNAs that negatively regulate gene expression via translational inhibition or mRNA degradation followed by protein synthesis repression. Many miRNAs are expressed in a tissue- and/or cell-specific manner and their expression patterns are reflective of underlying patho-physiologic processes. miRNAs can be detected in serum or in plasma in a remarkably stable form, making them attractive biomarkers for human diseases. This review describes the progress of identifying circulating miRNAs as novel biomarkers for diverse cardiovascular diseases, including acute myocardial infarction, heart failure, coronary artery disease, diabetes, stroke, essential hypertension, and acute pulmonary embolism. In addition, the origin and function and the different strategies to identify circulating miRNAs as novel biomarkers for cardiovascular diseases are also discussed. Rarely has an opportunity arisen to advance such new biology for the diagnosis of cardiac diseases. Content Type Journal Article Category Review Pages 1-11 DOI 10.1007/s00109-011-0840-5 Authors Jiahong Xu, Department of Cardiology, Tongji Hospital, Tongji University School of Medicine, 200065 Shanghai, China Jiangmin Zhao, Department of Radiology, Shanghai Third People’s Hospital, Shanghai Jiaotong University School of Medicine, 201900 Shanghai, China Graham Evan, Cardiovascular Institute, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02215, USA Chunyang Xiao, Cardiovascular Institute, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02215, USA Yan Cheng, Key Laboratory of Arrhythmias, Ministry of Education, China (Shanghai East Hospital, Tongji University School of Medicine), 200092 Shanghai, China Junjie Xiao, Key Laboratory of Arrhythmias, Ministry of Education, China (Shanghai East Hospital, Tongji University School of Medicine), 200092 Shanghai, China Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 7 Dec 2011 | 11:48 am CET

Erythropoietin attenuates the sequels of ischemic spinal cord injury with enhanced recruitment of CD34+ cells in mice Abstract Objective: Erythropoietin has been shown to promote tissue regeneration after ischemic injury in various organs. Here, we investigated whether Erythropoietin could ameliorate ischemic spinal cord injury in the mouse and sought an underlying mechanism. Methods: Spinal cord ischemia was developed by cross-clamping the descending thoracic aorta for 7- or 9-min in mice. Erythropoietin (5000IU/kg) or saline was administrated 30 min before aortic cross-clamping. Neurologic function was assessed using the paralysis score for 7 days after the operation. Spinal cords were histologically evaluated 2 and 7 days after the operation. Immunohistochemistry was used to detect CD34+ cells and the expression of brain-derived neurotrophic factor and vascular endothelial growth factor. Results: Each mouse exhibited either mildly impaired function or complete paralysis at day 2. Erythropoietin-treated mice with complete paralysis demonstrated significant improvement of neurologic function between day 2 and 7, compared to saline-treated mice with complete paralysis. Motor neurons in Erythropoietin-treated mice were more preserved at day 7 than those in saline-treated mice with complete paralysis. CD34+ cells in the lumbar spinal cord of Erythropoietin-treated mice were more abundant at day 2 than those of saline-treated mice. Brain-derived neurotrophic factor and vascular endothelial growth factor were markedly expressed in lumbar spinal cords in Erythropoietin-treated mice at day 7. Conclusion: Erythropoietin demonstrated neuroprotective effects in the ischemic spinal cord, improving neurologic function and attenuating motor neuron loss. These effects may have been mediated by recruited CD34+ cells, and enhanced expression of brain-derived neurotrophic factor and vascular endothelial growth factor. Quelle: Journal of Cellular and Molecular Medicine | 7 Dec 2011 | 10:02 am CET

Functional consequences of prolactin signaling in endothelial cells: a potential link with angiogenesis in pathophysiology? Abstract Prolactin is best known as the polypeptide anterior pituitary hormone, which regulates the development of the mammary gland. However, it became clear over the last decade that prolactin contributes to a broad range of pathologies, including breast cancer. Prolactin is also involved in angiogenesis via the release of pro-angiogenic factors by leukocytes and epithelial cells. However, whether prolactin also influences endothelial cells, and whether there are functional consequences of prolactin-induced signaling in the perspective of angiogenesis, remains so far elusive. In the present study, we show that prolactin induces phosphorylation of ERK1/2 and STAT5 and induces tube formation of endothelial cells on Matrigel. These effects are blocked by a specific prolactin receptor antagonist, del1-9-G129R-hPRL. Moreover, in an in vivo model of the chorioallantoic membrane of the chicken embryo, prolactin enhances vessel density and the tortuosity of the vasculature and pillar formation, which are hallmarks of intussusceptive angiogenesis. Interestingly, while prolactin has only little effect on endothelial cell proliferation, it markedly stimulates endothelial cell migration. Again, migration was reverted by del1-9-G129R-hPRL, indicating a direct effect of prolactin on its receptor. Immunohistochemistry and spectral imaging revealed that the prolactin receptor is present in the microvasculature of human breast carcinoma tissue. Altogether, these results suggest that prolactin may directly stimulate angiogenesis, which could be one of the mechanisms by which prolactin contributes to breast cancer progression, thereby providing a potential tool for intervention. Quelle: Journal of Cellular and Molecular Medicine | 1 Dec 2011 | 8:19 am CET

Generation of skeletal muscle cells from embryonic and induced pluripotent stem cells as an in vitro model and for therapy of muscular dystrophies Abstract Muscular dystrophies (MDs) are a heterogeneous group of inherited disorders characterized by progressive muscle wasting and weakness likely associated with exhaustion of muscle regeneration potential. At present, no cures or efficacious treatments are available for these diseases, but cell transplantation could be a potential therapeutic strategy. Transplantation of myoblasts using satellite cells or other myogenic cell populations has been attempted to promote muscle regeneration, based on the hypothesis that the donor cells repopulate the muscle and contribute to its regeneration. Embryonic stem cells (ESCs) and more recently induced pluripotent stem cells (iPSCs) could generate an unlimited source of differentiated cell types, including myogenic cells. Here we review the literature regarding the generation of myogenic cells considering the main techniques employed to date to elicit efficient differentiation of human and murine ESCs or iPSCs into skeletal muscle. We also critically analyse the possibility of using these cellular populations as an alternative source of myogenic cells for cell therapy of MDs. Quelle: Journal of Cellular and Molecular Medicine | 1 Dec 2011 | 8:19 am CET

Bone morphogenetic protein receptor 2 in patients with idiopathic portal hypertension Abstract Background. In idiopathic portal hypertension (IPH) typical vascular lesions are present in the branches of the portal vein or in the perisinusoidal area of the liver. Similar histological alterations have been reported in the pulmonary vasculature of patients with idiopathic pulmonary hypertension (IPAH). Since IPAH is associated with mutations of the BMPR2 gene, the aim of this study was to investigate whether this association might also be found in patients with IPH. Patients and Methods. Twenty three samples belonging to 21 unrelated caucasian patients with IPH followed in the hepatic hemodynamic laboratory of the Hospital Clinic in Barcelona were included in the study. All patients were studied for the entire open reading frame and splice site of the BMPR2 gene by direct sequencing and Multiple Ligation Probe Amplification (MLPA) in order to detect large deletions/duplications. Results. None of the 23 patients had pulmonary artery hypertension. Four patients presented one single nucleotide polymorphism (SNP) in intron 5, four patients had a SNP in exon 12 and a SNP in exon 1 was found in two cases. Two patients had both intron 5 and exon 12 polymorphisms. All SNPs were previously described. Except for these 3 SNPs, neither mutations nor rearrangements have been identified in the BMPR2 gene in this population. Conclusions. We did not detected mutations or rearrangements in the coding region of the BMPR2 gene in our patients with IPH. These findings suggest that, in contrast to IPAH, mutations in BMPR2 are not be involved in the pathogenesis of IPH. Quelle: Journal of Cellular and Molecular Medicine | 1 Dec 2011 | 8:19 am CET

Effects of valsartan on ventricular arrhythmia induced by programmed electrical stimulation in rats with myocardial infarction Abstract BACKGROUND The impact of Angiotensin II receptor blockers (ARBs) on electrical remodeling post myocardial infarction (MI) remains unclear. OBJECTIVES The purpose of the present study was to evaluate the effect of valsartan on incidence of ventricular arrhythmia induced by programmed electrical stimulation (PES) and potential link to changes of myocardial connexins (Cx) 43 expression and distribution in MI rats. METHODS Fifty-nine rats were randomly divided into three groups: Sham (n = 20), MI (n = 20) and MI+Val (20 mg/kg/day per gavage, n = 19). After 8 weeks, the incidence of PES induced ventricular tachycardia (VT) and fibrillation (VF) was compared among groups. mRNA and protein expressions of Cx43, angiotensin II type 1 receptor (AT1R) in the left ventricular border zone (BZ) and non-infarct zone (NIZ) were determined by Realtime-PCR and Western blot, respectively. Cx43 protein and collagen distribution were examined by immunohistochemistry in BZ and NIZ sections from MI hearts. RESULTS Valsartan effectively improved the cardiac function, reduced the prolonged QTc (163.7±3.7ms vs. 177.8±4.5ms, P<0.05) post MI and the incidence of VT or VF evoked by PES (21.1%vs. 55%, P<0.05). AT1R expression was significantly increased in BZ and NIZ sections post MI, which was downregulated by valsartan. The mRNA and protein expressions of Cx43 in BZ were significantly reduced post MI and upregulated by valsartan. Increased collagen deposition and reduced Cx43 expression in BZ post MI could be partly attenuated by Valsartan. CONCLUSIONS Valsartan reduced the incidence of PES induced ventricular arrhythmia, this effect was possibly through modulating the myocardial AT1R and Cx43 expression. Quelle: Journal of Cellular and Molecular Medicine | 30 Nov 2011 | 6:41 am CET

Repertoire of endothelial progenitor cells mobilized by femoral artery ligation – A nonhuman primate study Abstract To determine in the baboon model the identities and functional characteristics of endothelial progenitor cells mobilized in response to artery ligation, we collected peripheral blood mononuclear cells before and 3 days after a segment of femoral artery was removed. Our goal was to find EPC subpopulations with highly regenerative capacity. We identified 12 subpopulations of putative EPCs that were altered >1.75 fold; two subpopulations (CD146+/CD54-/CD45- at 6.63 fold, and CD146+/UEA-1-/CD45- at 12.21 fold) were dramatically elevated. To investigate the regenerative capacity of putative EPCs, we devised a new assay that maximally resembled their in vivo scenario, we purified CD34+ and CD146+ cells and co-cultured them with basal and mobilized peripheral blood mononuclear cells; both cell types took up Dil-LDL, but purified CD146+ cells exhibited accelerated differentiation by increasing expression of CD31 and CD144, and by exhibiting more active cord-like structure formation by comparison to the CD34+ subpopulation in a co-culture with mobilized PBMNCs. We demonstrate that ischemia due to vascular ligation mobilizes multiple types of cells with distinct roles. Baboon CD146+ cells exhibit higher reparative capacity than CD34+ cells, and thus are a potential source for therapeutic application. Quelle: Journal of Cellular and Molecular Medicine | 30 Nov 2011 | 6:41 am CET

Activation of the Prolyl-hydroxylase oxygen-sensing signal cascade leads to AMPK activation in cardiomyocytes Abstract Background- The proline hydroxylase domain-containing enzymes（PHD）act as cellular oxygen sensors and initiate a hypoxic signal cascade to induce a range of cellular responses to hypoxia especially in the aspect of energy and metabolic homeostasis regulation. AMP-activated protein kinase (AMPK) is recognized as a major energetic sensor and regulator of cardiac metabolism. However, the effect of PHD signal on AMPK has never been studied before. Methods and Results- A PHD inhibitor (PHI), dimethyloxaloylglycine, and PHD2-specific RNA interference (RNAi) have been used to activate PHD signaling in neonatal rat cardiomyocytes. Both PHI and PHD2-RNAi activated AMPK pathway in cardiomyocytes effectively. In addition, the increased glucose uptake during normoxia and enhanced myocyte viability during hypoxia induced by PHI pretreatment were abrogated substantially upon AMPK inhibition with an adenoviral vector expressing a dominant negative mutant of AMPK-alpha1. Furthermore, chelation of intracellular Ca2+ by BAPTA，inhibition of calmodulin-dependent kinase kinase (CaMKK) with STO-609, or RNAi mediated downregulation of CaMKK alpha inhibited PHI-induced AMPK activation significantly. In contrast, downregulation of LKB1 with adenoviruses expressing the dominant negative form did not affect PHI-induced AMPK activation. Conclusions- We establish for the first time that activation of PHD signal cascade can activate AMPK pathway mainly through a Ca2+/CaMKK-dependent mechanism in cardiomyocytes. Furthermore, activation of AMPK plays an essential role in hypoxic protective responses induced by PHI. Quelle: Journal of Cellular and Molecular Medicine | 30 Nov 2011 | 6:40 am CET

Mild hypoxia induced cardiomyocyte hypertrophy via up-regulation of HIF-1α-mediated TRPC signaling Abstract Hypoxia-inducible factor-1 alpha (HIF-1α) is a central transcriptional regulator of hypoxic response. The present study was designed to investigate the role of HIF-1α in mild hypoxia-induced cardiomyocytes hypertrophy and its underlying mechanism. Mild hypoxia (MH, 10% O2) caused hypertrophy in cultured neonatal rat cardiac myocytes, which was accompanied with increase of HIF-1α mRNA and accumulation of HIF-1α protein in nuclei. Transient receptor potential canonical (TRPC) channels including TRPC3 and TRPC6, except for TRPC1 were increased, and Ca2+-calcineurin signals were also enhanced in a time-dependent manner under MH condition. MH-induced cardiomyocytes hypertrophy, TRPC up-regulation and enhanced Ca2+-calcineurin signals were inhibited by a HIF-1α specific blocker, SC205346 (30 μM); whereas promoted by HIF-1α overexpression. Electrophysiological voltage-clamp demonstrated that DAG analogue, OAG (30 μM) induced TRPC current by as much as 170% in neonatal rat cardiomyocytes overexpressing HIF-1α compared to negative control. These results implicate that HIF-1α plays a key role in development of cardiac hypertrophy in responses to hypoxic stress. Its mechanism is associated with up-regulating TRPC3, TRPC6 expression, activating TRPC current and subsequently leading to enhanced Ca2+- calcineurin signals. Quelle: Journal of Cellular and Molecular Medicine | 30 Nov 2011 | 6:40 am CET

Endothelial fsh receptor in primary kidney cancer correlates with subsequent response to sunitinib Abstract Sunitinib is an antiangiogenic receptor tyrosine kinase inhibitor used to treat advanced metastatic renal cell carcinoma and other types of cancer. Sutent is effective in only approx. 70% of clear cell renal cell carcinoma (CCRCC) patients, has significant adverse side effects, and no method is available to predict which patients will not respond. Our purpose was to explore the possibility of introducing an effective prediction method based on a marker of the tumor vasculature, the Follicle Stimulating Hormone Receptor (FSHR). Fifty patients diagnosed with advanced metastatic CCRCC have been subjected to surgery for removal of the primary tumor and were subsequently treated with sunitinib. After three months of therapy the patients were categorized as “responsive”, “stable”, or “non-responsive” based on the RECIST guidelines. The blood vessel density and the percentage of FSHR-positive vessels were determined by immunofluorescence on sections from the primary tumors removed by surgery, prior to the sunitinib treatment. The percentage of FSHR-stained vessels was on average five fold higher for the patients that responded to the treatment in comparison with the stable group and almost eight fold higher than in the non-responsive group. The percentage allowed the detection of responders with 87–100% sensitivity and specificity. No significant differences were detected in the total density of vessels among the three groups. The data suggest that FSHR expression levels in the blood vessels of CCRCC primary tumors can be used to predict, with high sensitivity and specificity, the patients that will respond to sunitinib therapy. Quelle: Journal of Cellular and Molecular Medicine | 30 Nov 2011 | 6:40 am CET

Pravastatin-induced proangiogenic effects depend upon extracellular FGF-2 Abstract The HMG-CoA reductase inhibitors (statins) have been shown to exert several protective effects on the vasculature that are unrelated to changes in the cholesterol profile, and to induce angiogenesis. The proangiogenic effect exerted by statins has been attributed to the activation of the PI3K/Akt pathway in endothelial cells; however, it is unclear how statins activate this pathway. Pravastatin-mediated activation of Akt and MAPK occurs rapidly (within 10 min) and at low doses (0.01 μM). Here, we hypothesized that FGF-2 contributes to the proangiogenic effect of statins. We found that pravastatin, a hydrophilic statin, induced phosphorylation of the FGF receptor (FGFR) in human umbilical vein endothelial cells. SU5402, an inhibitor of FGFR, abolished pravastatin-induced PI3K/Akt and MAPK activity. Likewise, anti-FGF-2 function-blocking antibodies inhibited Akt and MAPK activity. Moreover, depletion of extracellular FGF-2 by heparin prevented pravastatin-induced phosphorylation of Akt and MAPK. Treatment with FGF-2 antibody inhibited pravastatin-enhanced endothelial cell proliferation, migration, and tube formation. These observations indicate that pravastatin exerts proangiogenic effects in endothelial cells depending upon the extracellular FGF-2. Quelle: Journal of Cellular and Molecular Medicine | 28 Nov 2011 | 10:41 am CET

Rapamycin inhibits transforming growth factor β induced peritoneal angiogenesis by blocking the secondary hypoxic response Abstract Patients with end stage kidney disease on peritoneal dialysis often develop progressive scarring of the peritoneal tissues. This manifests as submesothelial thickening and is associated with increased vascularization that leads to ultrafiltration dysfunction. Hypoxia induces a characteristic series of responses including angiogenesis and fibrosis. We investigated the role of hypoxia in peritoneal membrane damage. An adenovirus expressing transforming growth factor (TGF) β was used to induce peritoneal fibrosis. We evaluated the effect of the mTOR inhibitor rapamycin which has been previously shown to block hypoxia inducible factor (HIF) 1α. We also assessed the effect of HIF1α independently using an adenovirus expressing active HIF1α. To identify the TGFβ1 independent effects of HIF1α, we expressed HIF1α in the peritoneum of mice lacking the TGFβ signaling molecule Smad3. We demonstrate that TGFβ-induced fibroproliferative tissue is hypoxic. Rapamycin did not affect the early angiogenic response, but inhibited angiogenesis and submesothelial thickening 21 days after induction of fibrosis. In primary mesothelial cell culture, rapamycin had no effect on TGFβ induced vascular endothelial growth factor (VEGF) but did suppress hypoxia induced VEGF. HIF1α induced submesothelial thickening and angiogenesis in peritoneal tissue. The fibrogenic effects of HIF1α were Smad3 dependent. In summary, submesothelial hypoxia may be an important secondary factor which augments TGFβ induced peritoneal injury. The hypoxic response is mediated partly through HIF1α and the mTOR inhibitor rapamycin blocks the hypoxic induced angiogenic effects but does not affect the direct TGFβ mediated fibrosis and angiogenesis. Quelle: Journal of Cellular and Molecular Medicine | 28 Nov 2011 | 10:40 am CET

Pro-inflammatory phenotype of COPD fibroblasts not compatible with repair in COPD lung Abstract Chronic obstructive pulmonary disease (COPD) is characterized by loss of elastic fibers from small airways and alveolar walls, with the decrease in elastin increasing with disease severity. It is unclear why there is a lack of repair of elastic fibers. We have examined fibroblasts cultured from lung tissue from subjects with or without COPD to determine if the secretory profile explains lack of tissue repair. In this study, fibroblasts were cultured from lung parenchyma of patients with mild COPD (GOLD 1, n = 5), moderate-severe COPD (GOLD 2–3, n = 12), and controls (non-COPD, n = 5). Measurements were made of proliferation, senescence-associated beta-galactosidase-1, mRNA expression of IL-6, IL-8, MMP-1, tropoelastin and versican, and protein levels for IL-6, IL-8, PGE2, tropoelastin, insoluble elastin, and versican. GOLD 2–3 fibroblasts proliferated more slowly (p<0.01), had higher levels of senescence-associated beta-galactosidase-1 (p<0.001) than controls, and showed significant increases in mRNA and/or protein for IL-6 (p<0.05), IL-8 (p<0.01), MMP-1 (p<0.05), PGE2 (p<0.05), versican (p<0.05) and tropoelastin (p<0.05). mRNA expression and/or protein levels of tropoelastin (p<0.01), versican (p<0.05), IL-6 (p<0.05) and IL-8 (p<0.05) were negatively correlated with FEV1% of predicted. Insoluble elastin was not increased. In summary, fibroblasts from moderate-severe COPD subjects display a secretory phenotype with up-regulation of inflammatory molecules including the matrix proteoglycan versican, and increased soluble, but not insoluble, elastin. Versican inhibits assembly of tropoelastin into insoluble elastin and we conclude that the pro-inflammatory phenotype of COPD fibroblasts is not compatible with repair of elastic fibers. Quelle: Journal of Cellular and Molecular Medicine | 28 Nov 2011 | 10:39 am CET

The Akt1 isoform is an essential mediator of ischemic preconditioning Abstract PI3K-Akt pathway is essential for conferring cardioprotection in response to ischemic preconditioning (IPC) stimulus. However, the role of the individual Akt isoforms expressed in the heart in mediating the protective response to IPC is unknown. In this study we investigated the specific contribution of Akt1 and Akt2 in cardioprotection against ischemia-reperfusion (I-R) injury. Mice deficient in Akt1 or Akt2 were subjected to in vivo regional myocardial ischemia for 30 minutes followed by reperfusion for 2 hours with or without a prior IPC stimulus. Our results show that mice deficient in Akt1 were resistant to protection with either 1 or 3 cycles of IPC stimulus (42.7±6.5% control versus 38.5±1.9% 1xIPC, N = 6, NS; 41.4±6.3% control versus 32.4±3.2% 3xIPC, N = 10, NS). Western blot analysis, performed on heart samples taken from Akt1−/− mice subjected to IPC, revealed an impaired phosphorylation of GSK-3β, a downstream effector of Akt, as well as Erk1/2, the parallel component of the reperfusion injury salvage kinase pathway. Akt2−/− mice, which exhibit a diabetic phenotype, however, were amenable to protection with 3 but not 1 cycle of IPC (46.4±5.6% control versus 35.9±5.0% in 1xIPC, N = 6, NS; 47.0±6.0% control versus 30.8±3.3% in 3xIPC, N = 6; *P = 0.039). Akt1 but not Akt2 is essential for mediating a protective response to an IPC stimulus. Impaired activation of GSK-3β and Erk1/2 might be responsible for the lack of protective response to IPC in Akt1−/− mice. The rise in threshold for protection in Akt2−/− mice might be due to their diabetic phenotype. Quelle: Journal of Cellular and Molecular Medicine | 28 Nov 2011 | 10:36 am CET

Myelopoiesis in spleen producing distinct dendritic-like cells Abstract Dendritic cells (DC) represent a heterogeneous class of antigen presenting cells (APC). Previously we reported a distinct myeloid dendritic-like cell present in spleen, as an in vivo counterpart to cells produced in murine spleen long-term cultures (LTC-DC). These cells, named ‘L-DC’, were found to be functionally and phenotypically distinct from conventional (c)DC, plasmacytoid (p)DC and monocytes. These results suggested that spleen may represent a niche for development of L-DC from endogenous progenitors. Adult murine spleen has now been investigated for the presence of L-DC progenitors. Lineage-negative (Lin)−ckitlo and Lin−ckithi progenitor subsets were identified as candidate populations, and tested for ability to produce L-DC; in vitro upon co-culture with the spleen stromal line STX3, and in vivo after adoptive therapy into mice. Both subsets colonised STX3 stroma in vitro for L-DC production, indicating that they contained either a common or two distinct progenitors for L-DC. However, only the Lin−ckithi subset gave progeny cells after adoptive transfer into lethally irradiated mice. In vivo development was however multilineage and not restricted to L-DC development. Multilineage reconstitution reflects longterm reconstituting hematopoietic stem cells (LT-HSC), suggesting a close relationship between L-DC progenitors and LT-HSC. L-DC were however produced in vivo in much higher number than monocytes/macrophages and cDC, indicating the presence of a specific L-DC progenitor within the Lin−ckithi subset. A model is advanced for development of L-DC directly from hematopoietic progenitors in spleen and dependent on the spleen microenvironment. Quelle: Journal of Cellular and Molecular Medicine | 28 Nov 2011 | 10:36 am CET

Altered apoptosis regulation in kufor-rakeb syndrome patients with mutations in the atp13a2 gene Abstract ATP13A2 gene encodes for a protein of the group 5 P-type ATPase family. ATP13A2 mutations are responsible for Kufor-Rakeb syndrome, a rare autosomal recessive juvenile parkinsonism characterized by the subacute onset of extrapyramidal, pyramidal and cognitive dysfunction with secondary nonresponsiveness to Levodopa. FBXO7 protein is an F-box-containing protein. Recessive FBXO7 mutations are responsible for PARK15, a rare juvenile parkinsonism characterized by progressive neurodegeneration with extrapyramidal and pyramidal system involvement. Our aim was to evaluate apoptosis in cells from two KRS siblings carrying a homozygous ATP13A2 mutation and a heterozygous FBXO7 mutation. We also analysed apoptosis in the patients’ healthy parents. Peripheral blood lymphocytes from the KRS patients and parents were exposed to 2-deoxy-D-ribose; apoptosis was analysed by flow cytometry and by fluorescence microscopy. Apoptosis was much higher in lymphocytes from the KRS patients and parents than in controls, both in standard conditions and after induction with a pro-apoptotic stimulus. The lack of correlation between increased apoptosis and the presence of the mutated FBXO7 gene rules out the involvement of FBXO7 in apoptosis regulation. The altered apoptotic pattern of subjects with mutated ATP13A2 suggests a correlation between apoptosis alteration and the mutated ATP13A2 protein. We hypothesize that ATP13A2 mutations may compromise protein function, disrupting cell cation balance and rendering cells prone to apoptosis. However, the deregulation of apoptosis in Kufor-Rakeb syndrome patients displaying different disease severity suggested that the altered apoptotic pathway probably does not have a pathogenetic role in Kufor-Rakeb syndrome by itself. Quelle: Journal of Cellular and Molecular Medicine | 28 Nov 2011 | 10:35 am CET

Aberrant epigenetic regulation of bromodomain Brd4 in human colon cancer Abstract   The bromodomain protein BRD4 is involved in cell proliferation and cell cycle progression, primarily through its role in acetylated chromatin-dependent regulation of transcription at targeted loci. Here, we show that BRD4 is frequently downregulated by aberrant promoter hypermethylation in human colon cancer cell lines and primary tumors. Ectopic re-expression of BRD4 in these colon cancer cell lines markedly reduced in vivo tumor growth, suggesting a role of BRD4 in human colon cancer. Content Type Journal Article Category Original Article Pages 1-9 DOI 10.1007/s00109-011-0837-0 Authors R. M. Rodriguez, Cancer Epigenetics Laboratory, Instituto Universitario de Oncología del Principado de Asturias (IUOPA), HUCA, Universidad de Oviedo, Oviedo, Spain C. Huidobro, Cancer Epigenetics Laboratory, Instituto Universitario de Oncología del Principado de Asturias (IUOPA), HUCA, Universidad de Oviedo, Oviedo, Spain R. G. Urdinguio, Cancer Epigenetics Laboratory, Instituto Universitario de Oncología del Principado de Asturias (IUOPA), HUCA, Universidad de Oviedo, Oviedo, Spain C. Mangas, Cancer Epigenetics Laboratory, Instituto Universitario de Oncología del Principado de Asturias (IUOPA), HUCA, Universidad de Oviedo, Oviedo, Spain B. Soldevilla, Department of Medical Oncology, Hospital Universitario Puerta de Hierro, Universidad Autónoma de Madrid, C/Manuel de Falla 1, Majadahonda, Madrid, Spain G. Domínguez, Department of Medical Oncology, Hospital Universitario Puerta de Hierro, Universidad Autónoma de Madrid, C/Manuel de Falla 1, Majadahonda, Madrid, Spain F. Bonilla, Department of Medical Oncology, Hospital Universitario Puerta de Hierro, Universidad Autónoma de Madrid, C/Manuel de Falla 1, Majadahonda, Madrid, Spain A. F. Fernandez, Cancer Epigenetics Laboratory, Instituto Universitario de Oncología del Principado de Asturias (IUOPA), HUCA, Universidad de Oviedo, Oviedo, Spain M. F. Fraga, Cancer Epigenetics Laboratory, Instituto Universitario de Oncología del Principado de Asturias (IUOPA), HUCA, Universidad de Oviedo, Oviedo, Spain Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 26 Nov 2011 | 5:48 pm CET

Differentiation of multiple types of pancreatico-biliary tumors by molecular analysis of clinical specimens Abstract   Timely and accurate diagnosis of pancreatic ductal adenocarcinoma (PDAC) is critical in order to provide adequate treatment to patients. However, the clinical signs and symptoms of PDAC are shared by several types of malignant or benign tumors which may be difficult to differentiate from PDAC with conventional diagnostic procedures. Among others, these include ampullary cancers, solid pseudopapillary tumors, and adenocarcinomas of the distant bile duct, as well as inflammatory masses developing in chronic pancreatitis. Here, we report an approach to accurately differentiate between these different types of pancreatic masses based on molecular analysis of biopsy material. A total of 156 bulk tissue and fine needle aspiration biopsy samples were analyzed using a dedicated diagnostic cDNA array and a composite classification algorithm developed based on linear support vector machines. All five histological subtypes of pancreatic masses were clearly separable with 100% accuracy when using all 156 individual samples for classification. Generalized performance of the classification system was tested by 10 × 10-fold cross validation (100 test runs). Correct classification into the five diagnostic groups was demonstrated for 81.5% of 1,560 test set predictions. Performance increased to 85.3% accuracy when PDAC and distant bile duct carcinomas were combined in a single diagnostic class. Importantly, overall sensitivity of detection of malignant disease was 92.2%. The molecular diagnostic approach presented here is suitable to significantly aid in the differential diagnosis of undetermined pancreatic masses. To our knowledge, this is the first study reporting accurate differentiation between several types of pancreatico-biliary tumors in a single molecular analytical procedure. Content Type Journal Article Category Original Article Pages 1-8 DOI 10.1007/s00109-011-0832-5 Authors Thomas M. Gress, Division of Gastroenterology, University Hospital, Philipps-Universitaet Marburg, Marburg, Germany Hans A. Kestler, Internal Medicine I, University Hospital Ulm, Ulm, Germany Ludwig Lausser, Research group of Bioinformatics and Systems Biology, Institute of Neural Information Processing, University of Ulm, Ulm, Germany Lisa Fiedler, Division of Gastroenterology, University Hospital, Philipps-Universitaet Marburg, Marburg, Germany Bence Sipos, Institute of Pathology, University Hospital Tübingen, Tübingen, Germany Christoph W. Michalski, Department of Surgery, Technical University of Munich, Munich, Germany Jens Werner, Department of Surgery, University of Heidelberg, Heidelberg, Germany Nathalia Giese, Department of Surgery, University of Heidelberg, Heidelberg, Germany Aldo Scarpa, Department of Pathology, University of Verona, Verona, Italy Malte Buchholz, Division of Gastroenterology, University Hospital, Philipps-Universitaet Marburg, Marburg, Germany Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 25 Nov 2011 | 6:59 pm CET

Erbin inhibits TGF-β1-induced EMT in renal tubular epithelial cells through an ERK-dependent pathway Abstract   Epithelial-to-mesenchymal transition (EMT) plays a crucial role in the progression of renal interstitial fibrosis, which finally leads to renal failure. Erbin, a member of LAP family, is recently reported to inhibit Smads and ERK pathway which are two important types of intracellular signaling involved in TGF-β1-induced EMT. However, the role of Erbin in the regulation of EMT and the underlying mechanisms remain to be fully understood. To that end, we aimed to evaluate the expression of Erbin in renal interstitial fibrosis and the potential role of Erbin in tubular EMT stimulated by TGF-β1. In this study we demonstrated that the expression of Erbin was upregulated in the tubular epithelia of 5/6-nephrectomized rats. We also showed here that TGF-β1 upregulated Erbin expression in NRK52E cells during their EMT phenotype acquisition. Importantly, elevated expression of Erbin inhibited ERK signaling and partial reversed EMT stimulated by TGF-β1. In the mean time, reducing Erbin expression enhanced ERK phosphorylation, promoted the E-cadherin suppression, and induced α-SMA expression and fibronection secretion in response to TGF-β1, which could be rescued if cells were treated with the inhibitor of MEK1/2 U0126. However, in the absence of TGF-β1, Erbin failed to affect ERK activation and EMT process. These results suggest that Erbin is a negative feedback molecule induced by TGF-β1 and inhibits TGF-β1-induced EMT via ERK signaling pathway. Content Type Journal Article Category Original Article Pages 1-12 DOI 10.1007/s00109-011-0833-4 Authors Qiaodan Zhou, Division of Nephrology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, 430030 Hubei, People’s Republic of China Rui Zeng, Division of Nephrology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, 430030 Hubei, People’s Republic of China Chuou Xu, Division of Nephrology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, 430030 Hubei, People’s Republic of China Lili Liu, Division of Nephrology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, 430030 Hubei, People’s Republic of China Lin Chen, Department of Hepatic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, 430030 Hubei, People’s Republic of China Pei Kou, Division of Nephrology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, 430030 Hubei, People’s Republic of China Guangchang Pei, Division of Nephrology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, 430030 Hubei, People’s Republic of China Shoujun Bai, Division of Nephrology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, 430030 Hubei, People’s Republic of China Yamin Zhang, Division of Nephrology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, 430030 Hubei, People’s Republic of China Caixia Li, Division of Nephrology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, 430030 Hubei, People’s Republic of China Song Rong, Department of Nephrology, Hannover Medical School, Hannover, Germany Min Han, Division of Nephrology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, 430030 Hubei, People’s Republic of China Gang Xu, Division of Nephrology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Ave., Wuhan, 430030 Hubei, People’s Republic of China Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 24 Nov 2011 | 6:49 pm CET

Homeobox gene Distal-Less 3 is a regulator of villous cytotrophoblast differentiation and its expression is increased in human idiopathic foetal growth restriction Abstract   Human idiopathic foetal growth restriction (FGR) is frequently associated with placental insufficiency. In our previous studies, we have reported the isolation and characterisation of the homeobox gene Distal-less 3 (DLX3) in the human placenta. In this study, we have investigated the level of DLX3 expression in idiopathic FGR-affected placentae and determined its functional role in villous trophoblast differentiation. FGR-affected placentae (n = 25) were collected based on well-defined clinical criteria and matched for gestation with control uncomplicated pregnancies (n = 25). Real-time polymerase chain reaction and immunoblotting showed increased DLX3 mRNA and protein expression in FGR-affected placentae compared with gestation-matched controls. Qualitative immunohistochemistry revealed DLX3 localisation in the syncytiotrophoblast, cytotrophoblasts and endothelial cells surrounding the foetal capillaries in both FGR-affected and control placentae. Down-regulation of DLX3 in primary villous trophoblast cells and a trophoblast-derived cell line showed decreased expression of differentiation markers, 3βHSD, βhCG and syncytin. Therefore, we conclude that increased DLX3 expression in FGR may contribute to trophoblast dysfunction observed in FGR. Content Type Journal Article Category Original Article Pages 1-12 DOI 10.1007/s00109-011-0836-1 Authors Amy Chui, Department of Perinatal Medicine, Pregnancy Research Centre, Royal Women’s Hospital, Parkville, VIC 3052, Australia Charmaine Tay, Department of Perinatal Medicine, Pregnancy Research Centre, Royal Women’s Hospital, Parkville, VIC 3052, Australia Melanie Cocquebert, INSERM, U767, Faculté des Sciences Pharmaceutiques et Biologiques, Paris, 75006 France Penelope Sheehan, Department of Perinatal Medicine, Pregnancy Research Centre, Royal Women’s Hospital, Parkville, VIC 3052, Australia Niroshani A. Pathirage, Department of Perinatal Medicine, Pregnancy Research Centre, Royal Women’s Hospital, Parkville, VIC 3052, Australia Susan Donath, Clinical Epidemiology and Biostatistics Unit, Murdoch Children’s Research Institute, Royal Children’s Hospital, Parkville, VIC 3052, Australia Thierry Fournier, INSERM, U767, Faculté des Sciences Pharmaceutiques et Biologiques, Paris, 75006 France Josette Badet, INSERM, U767, Faculté des Sciences Pharmaceutiques et Biologiques, Paris, 75006 France Daniele Evain-Brion, INSERM, U767, Faculté des Sciences Pharmaceutiques et Biologiques, Paris, 75006 France Shaun P. Brennecke, Department of Perinatal Medicine, Pregnancy Research Centre, Royal Women’s Hospital, Parkville, VIC 3052, Australia Bill Kalionis, Department of Perinatal Medicine, Pregnancy Research Centre, Royal Women’s Hospital, Parkville, VIC 3052, Australia Padma Murthi, Department of Perinatal Medicine, Pregnancy Research Centre, Royal Women’s Hospital, Parkville, VIC 3052, Australia Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 23 Nov 2011 | 6:42 pm CET

Rapamycin induces glucose intolerance in mice by reducing islet mass, insulin content, and insulin sensitivity Abstract   Rapamycin, a specific inhibitor for mTOR complex 1, is an FDA-approved immunosuppressant for organ transplant. Recent developments have raised the prospect of using rapamycin to treat cancer or diabetes and to delay aging. It is therefore important to assess how rapamycin treatment affects glucose homeostasis. Here, we show that the same rapamycin treatment reported to extend mouse life span significantly impaired glucose homeostasis of aged mice. Moreover, rapamycin treatment of lean C57B/L6 mice reduced glucose-stimulated insulin secretion in vivo and ex vivo as well as the insulin content and beta cell mass of pancreatic islets. Confounding the diminished capacity for insulin release, rapamycin decreased insulin sensitivity. The multitude of rapamycin effects thus all lead to glucose intolerance. As our findings reveal that chronic rapamycin treatment could be diabetogenic, monitoring glucose homeostasis is crucial when using rapamycin as a therapeutic as well as experimental reagent. Content Type Journal Article Category Original Article Pages 1-11 DOI 10.1007/s00109-011-0834-3 Authors Shi-Bing Yang, Howard Hughes Medical Institute, Departments of Physiology, Biochemistry and Biophysics, University of California, San Francisco, 1550, 4th Street, San Francisco, CA 94158, USA Hye Young Lee, Howard Hughes Medical Institute, Departments of Physiology, Biochemistry and Biophysics, University of California, San Francisco, 1550, 4th Street, San Francisco, CA 94158, USA David Matthew Young, Howard Hughes Medical Institute, Departments of Physiology, Biochemistry and Biophysics, University of California, San Francisco, 1550, 4th Street, San Francisco, CA 94158, USA An-Chi Tien, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Howard Hughes Medical Institute, University of California, San Francisco, 35 Medical Center Way, San Francisco, CA 94143, USA Ashley Rowson-Baldwin, Howard Hughes Medical Institute, Departments of Physiology, Biochemistry and Biophysics, University of California, San Francisco, 1550, 4th Street, San Francisco, CA 94158, USA Yu Yu Shu, Howard Hughes Medical Institute, Departments of Physiology, Biochemistry and Biophysics, University of California, San Francisco, 1550, 4th Street, San Francisco, CA 94158, USA Yuh Nung Jan, Howard Hughes Medical Institute, Departments of Physiology, Biochemistry and Biophysics, University of California, San Francisco, 1550, 4th Street, San Francisco, CA 94158, USA Lily Yeh Jan, Howard Hughes Medical Institute, Departments of Physiology, Biochemistry and Biophysics, University of California, San Francisco, 1550, 4th Street, San Francisco, CA 94158, USA Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 21 Nov 2011 | 7:04 pm CET

Leukocyte integrin activation and deactivation: novel mechanisms of balancing inflammation Abstract   Leukocyte recruitment into tissue forms the basis of immune surveillance and direct immune defense. It proceeds in a cascade-like fashion. The first contact of leukocytes with the endothelium is mediated by selectins and their counter receptors, followed by rolling and integrin-mediated arrest. While rolling, neutrophils collect different inflammatory signals which can activate several signaling pathways leading to leukocyte adhesion to the endothelium and transmigration through the blood vessel wall into the inflamed tissue. Whereas inflammatory reactions are beneficial and necessary for host defense, they need to be balanced and controlled to prevent harmful consequences and tissue destruction. In this article, we discuss the different signaling pathways that ensure rapid and efficient integrin activation on leukocytes. In addition, we report on a recently identified novel endogenous mechanism that counteracts and balances integrin activation, thereby limiting leukocyte recruitment and the extent of inflammation. Further investigation of this new mechanism may allow providing new approaches for the development of the next generation of anti-inflammatory drugs. Content Type Journal Article Category Review Pages 1-7 DOI 10.1007/s00109-011-0835-2 Authors Alexander Zarbock, Department of Anesthesiology and Intensive Care Medicine, University of Münster, Albert-Schweitzer Str. 33, 48149 Münster, Germany Tibor Kempf, Molecular and Translational Cardiology, Department of Cardiology and Angiology, Hannover Medical School, Hannover, Germany Kai C. Wollert, Molecular and Translational Cardiology, Department of Cardiology and Angiology, Hannover Medical School, Hannover, Germany Dietmar Vestweber, Max-Planck Institute for Molecular Biomedicine, Münster, Germany Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 18 Nov 2011 | 6:32 pm CET

Salivary transcriptomic biomarkers for detection of ovarian cancer: for serous papillary adenocarcinoma Abstract   Ovarian cancer is the most lethal gynecological cancer due to lack of clear symptom and reliable screening biomarker in the early stage. The capability to detect the initiation of malignancy with a sensitive and effective approach is one of the most desirable goals for ovarian cancer therapy. In this study, we spearheaded noninvasive detection of ovarian cancer by salivary transcriptomic biomarkers, and evaluated the clinical utilities of discovered biomarkers using a clinical case–control study. To find salivary mRNA biomarkers, salivary transcriptomes in 11 ovarian cancer patients and 11 matched controls were profiled by Affymetrix HG-U133-Plus-2.0 array. The biomarker candidates selected from the microarray results were then subjected to clinical validation by RT-qPCR using an independent sample cohort including 21 ovarian cancer patients and 35 healthy controls. Seven downregulated mRNA biomarkers were validated. The logistic regression model revealed the combination of five validated biomarkers (AGPAT1, B2M, BASP2, IER3, and IL1B) can significantly discriminate ovarian cancer patients (n = 21) from the healthy controls (n = 35), yielding a receiver operating characteristic plot, area under the curve value of 0.909 with 85.7% sensitivity and 91.4% specificity. In summary, we have demonstrated that the RNA signatures in saliva could serve as biomarkers for detection of ovarian cancer with high sensitivity and specificity. This emerging approach with high-throughput, noninvasive, and effective advantages provides a feasible means for detection of systemic cancer, and opens a new avenue for early disease detection. Content Type Journal Article Category Original Article Pages 1-8 DOI 10.1007/s00109-011-0829-0 Authors Yu-Hsiang Lee, Graduate Institute of Biomedical Engineering, National Central University, Jhongli City, 32001 Taiwan, Republic of China Jae Hoon Kim, Department of Obstetrics and Gynecology, Gangnam Biomedical Research Center Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, South Korea Hui Zhou, UCLA School of Dentistry and Dental Research Institute, 73-017 CHS, 10833 Le Conte Ave, Los Angeles, CA 90095, USA Bo Wook Kim, Tissue Array Research Program, Laboratory of Pathology, National Cancer Institute, Bethesda, MA, USA David T. Wong, UCLA School of Dentistry and Dental Research Institute, 73-017 CHS, 10833 Le Conte Ave, Los Angeles, CA 90095, USA Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 18 Nov 2011 | 7:58 am CET

Comparative kinome analysis to identify putative colon tumor biomarkers Abstract   Kinase domains are the type of protein domain most commonly found in genes associated with tumorigenesis. Because of this, the human kinome (the protein kinase component of the genome) represents a promising source of cancer biomarkers and potential targets for novel anti-cancer therapies. Alterations in the human colon kinome during the progression from normal colon (NC) through adenoma (AD) to adenocarcinoma (AC) were investigated using integrated transcriptomic and proteomic datasets. Two hundred thirty kinase genes and 42 kinase proteins showed differential expression patterns (fold change ≥ 1.5) in at least one tissue pair-wise comparison (AD vs. NC, AC vs. NC, and/or AC vs. AD). Kinases that exhibited similar trends in expression at both the mRNA and protein levels were further analyzed in individual samples of NC (n = 20), AD (n = 39), and AC (n = 24) by quantitative reverse transcriptase PCR. Individual samples of NC and tumor tissue were distinguishable based on the mRNA levels of a set of 20 kinases. Altered expression of several of these kinases, including chaperone activity of bc1 complex-like (CABC1) kinase, bromodomain adjacent to zinc finger domain protein 1B (BAZ1B) kinase, calcium/calmodulin-dependent protein kinase type II subunit delta (CAMK2D), serine/threonine-protein kinase 24 (STK24), vaccinia-related kinase 3 (VRK3), and TAO kinase 3 (TAOK3), has not been previously reported in tumor tissue. These findings may have diagnostic potential and may lead to the development of novel targeted therapeutic interventions for colorectal cancer. Content Type Journal Article Category Original Article Pages 1-10 DOI 10.1007/s00109-011-0831-6 Authors Ewa E. Hennig, Department of Gastroenterology and Hepatology, Medical Center for Postgraduate Education, Warsaw, Poland Michal Mikula, Department of Oncological Genetics, The Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland Tymon Rubel, Institute of Radioelectronics, Warsaw University of Technology, Warsaw, Poland Michal Dadlez, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland Jerzy Ostrowski, Department of Gastroenterology and Hepatology, Medical Center for Postgraduate Education, Warsaw, Poland Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 18 Nov 2011 | 7:58 am CET

Intermittent hypoxia activates temporally coordinated transcriptional programs in visceral adipose tissue Abstract   Obstructive sleep apnea (OSA) is a prevalent disorder characterized by intermittent hypoxia (IH) during sleep. OSA is strongly associated with obesity and dysregulation of metabolism—yet the molecular pathways linking the effects of IH on adipocyte biology remain unknown. We hypothesized that exposure to IH would activate distinct, time-dependent transcriptional programs in visceral adipose tissue of mice. We exposed 36 mice to IH or normoxia for up to 13 days. We transcriptionally profiled visceral fat tissue harvested from the animals and performed functional enrichment and network analysis on differentially expressed genes. We identified over 3,000 genes with significant expression patterns during the time course of IH exposure. The most enriched pathways mapped to metabolic processes, mitochondrion, and oxidative stress responses. We confirmed the pathophysiological relevance of these findings by demonstrating that mice exposed to chronic IH developed dyslipidemia and underwent significant lipid and protein oxidation within their visceral adipose depots. We applied gene–gene interaction network analysis to identify critical controllers of IH-induced transcriptional programs in adipocytes—these network hubs represent putative targets to modulate the effects of chronic IH on adipose tissue. Our approach to integrate computational methods with gene expression profiling of visceral fat tissue during IH exposure shows promise in helping unravel the mechanistic links between OSA and adipocyte biology. Content Type Journal Article Category Original Article Pages 1-11 DOI 10.1007/s00109-011-0830-7 Authors Sina A. Gharib, Center for Lung Biology, Department of Medicine, University of Washington, Seattle, WA, USA Abdelnaby Khalyfa, Department of Pediatrics, University of Chicago, 5721 S. Maryland Avenue, MC 8000, Suite K-160, Chicago, IL, USA Amal Abdelkarim, Department of Pediatrics, University of Chicago, 5721 S. Maryland Avenue, MC 8000, Suite K-160, Chicago, IL, USA Vijay Ramesh, Department of Pediatrics, University of Chicago, 5721 S. Maryland Avenue, MC 8000, Suite K-160, Chicago, IL, USA Mohamed Buazza, University of Louisville, Louisville, KY, USA Navita Kaushal, Department of Pediatrics, University of Chicago, 5721 S. Maryland Avenue, MC 8000, Suite K-160, Chicago, IL, USA Bharat Bhushan, Department of Pediatrics, University of Chicago, 5721 S. Maryland Avenue, MC 8000, Suite K-160, Chicago, IL, USA David Gozal, Department of Pediatrics, University of Chicago, 5721 S. Maryland Avenue, MC 8000, Suite K-160, Chicago, IL, USA Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 15 Nov 2011 | 5:48 pm CET

Suppression of antigen-specific CD4+ T cell activation by SRA/CD204 through reducing the immunostimulatory capability of antigen-presenting cell Abstract   Pattern recognition scavenger receptor SRA/CD204, primarily expressed on specialized antigen-presenting cells (APCs), including dendritic cells (DCs) and macrophages, has been implicated in multiple physiological and pathological processes, including atherosclerosis, Alzheimer’s disease, endotoxic shock, host defense, and cancer development. SRA/CD204 was also recently shown to function as an attenuator of vaccine response and antitumor immunity. Here, we, for the first time, report that SRA/CD204 knockout (SRA−/−) mice developed a more robust CD4+ T cell response than wild-type mice after ovalbumin immunization. Splenic DCs from the immunized SRA−/− mice were much more efficient than those from WT mice in stimulating naïve OT-II cells, indicating that the suppressive activity of SRA/CD204 is mediated by DCs. Strikingly, antigen-exposed SRA−/− DCs with or without lipopolysaccharide treatment exhibited increased T-cell-stimulating activity in vitro, which was independent of the classical endocytic property of the SRA/CD204. Additionally, absence of SRA/CD204 resulted in significantly elevated IL12p35 expression in DCs upon CD40 ligation plus interferon gamma (IFN-γ) stimulation. Molecular studies reveal that SRA/CD204 inhibited the activation of STAT1, mitogen activated protein kinase p38, and nuclear factor-kappa B signaling activation in DCs treated with anti-CD40 antibodies and IFN-γ. Furthermore, splenocytes from the generated SRA−/− OT-II mice showed heightened proliferation upon stimulation with OVA protein or MHC-II-restricted OVA323–339 peptide compared with cells from the SRA+/+ OT-II mice. These results not only establish a new role of SRA/CD204 in limiting the intrinsic immunogenicity of APCs and CD4+ T cell activation but also provide additional insights into the molecular mechanisms involved in the immune suppression by this molecule. Content Type Journal Article Category Original Article Pages 1-14 DOI 10.1007/s00109-011-0828-1 Authors Huanfa Yi, Department of Human & Molecular Genetics, Virginia Commonwealth University School of Medicine, PO Box 980033, Richmond, VA 23298, USA Daming Zuo, Department of Immunology, Southern Medical University, Guangzhou, 510515 China Xiaofei Yu, Department of Human & Molecular Genetics, Virginia Commonwealth University School of Medicine, PO Box 980033, Richmond, VA 23298, USA Fanlei Hu, Department of Human & Molecular Genetics, Virginia Commonwealth University School of Medicine, PO Box 980033, Richmond, VA 23298, USA Masoud H. Manjili, Department of Microbiology & Immunology, Virginia Commonwealth University, Richmond, VA 23298, USA Zhengliang Chen, Department of Immunology, Southern Medical University, Guangzhou, 510515 China John R. Subjeck, Department of Cellular Stress Biology, Roswell Park Cancer Institute, Buffalo, NY 14263, USA Xiang-Yang Wang, Department of Human & Molecular Genetics, Virginia Commonwealth University School of Medicine, PO Box 980033, Richmond, VA 23298, USA Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 14 Nov 2011 | 5:51 pm CET

Induction of primitive pigment cell differentiation by visible light (helium–neon laser): a photoacceptor-specific response not replicable by UVB irradiation Abstract   Solar lights encompass ultraviolet (UV), visible, and infrared spectrum. Most previous studies focused on the harmful UV effects, and the biologic effects of lights at other spectrums remained unclear. Recently, lights at visible region have been used for regenerative purposes. Using the process of vitiligo repigmentation as a research model, we focused on elucidating the pro-differentiation effects induced by visible light. We first showed that helium–neon (He–Ne) laser (632.8 nm) irradiation stimulated differentiation of primitive pigment cells, an effect not replicable by UVB treatment even at high and damaging doses. In addition, significant increases of mitochondrial DNA copy number and the regulatory genes for mitochondrial biogenesis were induced by He–Ne laser irradiation. Mechanistically, we demonstrated that He–Ne laser initiated mitochondrial retrograde signaling via a Ca2+-dependent cascade. The impact on cytochrome c oxidase within the mitochondria is responsible for the efficacy of He–Ne laser in promoting melanoblast differentiation. Taken together, we propose that visible lights from the sun provide important environmental cues for the relatively quiescent stem or primitive cells to differentiate. In addition, our results also indicate that visible light may be used for regenerative medical purposes involving stem cells. Content Type Journal Article Category Original Article Pages 1-10 DOI 10.1007/s00109-011-0822-7 Authors Cheng-Che E. Lan, Department of Dermatology, Kaohsiung Medical University Hospital, Faculty of Medicine, College of Medicine, and Center of Excellence for Environmental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, Republic of China Shi-Bei Wu, Institutes of Biochemistry and Molecular Biology, National Yang-Ming University, No. 155, Sec. 2, Li-Nong St., Taipei, Taipei, Taiwan, Republic of China Ching-Shuang Wu, Department of Medical Laboratory Science and Biotechnology, College of Health Science, Kaohsiung Medical University, Kaohsiung, Taiwan, Republic of China Yi-Chun Shen, Department of Dermatology, Kaohsiung Medical University Hospital, Faculty of Medicine, College of Medicine, and Center of Excellence for Environmental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, Republic of China Tzu-Ying Chiang, Department of Dermatology, Kaohsiung Medical University Hospital, Faculty of Medicine, College of Medicine, and Center of Excellence for Environmental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, Republic of China Yau-Huei Wei, Institutes of Biochemistry and Molecular Biology, National Yang-Ming University, No. 155, Sec. 2, Li-Nong St., Taipei, Taipei, Taiwan, Republic of China Hsin-Su Yu, Department of Dermatology, Kaohsiung Medical University Hospital, Faculty of Medicine, College of Medicine, and Center of Excellence for Environmental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, Republic of China Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 8 Nov 2011 | 8:13 pm CET

A novel hybrid promoter responsive to pathophysiological and pharmacological regulation Abstract   The aim of this study was to construct a promoter containing DNA motifs for an endogenous transcription factor associated with inflammation along with motifs for pharmacological regulation factors. We demonstrate in transfected cells that expression of a gene of interest is induced by hypoxic conditions or through pharmacological induction, and also show pharmacological repression. In vivo studies utilised electroporation of plasmid to mouse paws, a delivery method shown to be effective by bioluminescence imaging. For gene therapy, the promoter was used to drive expression of IL-1Ra in a paw inflammation model with therapeutic effect observed which was further enhanced when the promoter was additionally induced with a pharmacological activator. One of the most important observations from this study was that promoter induction by hypoxia or inflammation could be prevented by the pharmacological repressor in the absence of doxycycline. These studies demonstrate that hybrid promoters enable pharmacological adjustment to the pathophysiological level of gene expression and, importantly, that they allow termination of gene expression even in the presence of pathophysiological stimuli. Content Type Journal Article Category Original Article Pages 1-11 DOI 10.1007/s00109-011-0826-3 Authors Maria C. Subang, Bone and Joint Research Unit, Barts and The London School of Medicine and Dentistry, William Harvey Research Institute, Queen Mary University of London, Charterhouse Square, London, EC1M 6BQ UK Rewas Fatah, Bone and Joint Research Unit, Barts and The London School of Medicine and Dentistry, William Harvey Research Institute, Queen Mary University of London, Charterhouse Square, London, EC1M 6BQ UK Carly Bright, Bone and Joint Research Unit, Barts and The London School of Medicine and Dentistry, William Harvey Research Institute, Queen Mary University of London, Charterhouse Square, London, EC1M 6BQ UK Patricia Blanco, Fundacion Instituto Leloir, Av. Patricias Argentinas 435, Buenos Aires, Argentina Mariana Berenstein, Fundacion Instituto Leloir, Av. Patricias Argentinas 435, Buenos Aires, Argentina Ying Wu, Bone and Joint Research Unit, Barts and The London School of Medicine and Dentistry, William Harvey Research Institute, Queen Mary University of London, Charterhouse Square, London, EC1M 6BQ UK Osvaldo L. Podhajcer, Fundacion Instituto Leloir, Av. Patricias Argentinas 435, Buenos Aires, Argentina Paul G. Winyard, Institute of Biomedical and Clinical Science, Peninsula Medical School, University of Exeter, St Luke’s Campus, Exeter, UK Yuti Chernajovsky, Bone and Joint Research Unit, Barts and The London School of Medicine and Dentistry, William Harvey Research Institute, Queen Mary University of London, Charterhouse Square, London, EC1M 6BQ UK David Gould, Bone and Joint Research Unit, Barts and The London School of Medicine and Dentistry, William Harvey Research Institute, Queen Mary University of London, Charterhouse Square, London, EC1M 6BQ UK Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 8 Nov 2011 | 8:12 pm CET

Cell-derived microparticles in atherosclerosis: biomarkers and targets for pharmacological modulation? Abstract Cardiovascular diseases remain an important cause of morbi-mortality. Atherosclerosis, which predisposes to cardiovascular disorders such as myocardial infarction and stroke, develops silently over several decades. Identification of circulating biomarkers to evaluate cardiovascular event risk and pathology prognosis is of particular importance. Microparticles (MPs) are small vesicles released from cells upon apoptosis or activation. MPs are present in blood of healthy individuals. Studies showing a modification of their concentrations in patients with cardiovascular risk factors and after cardiovascular events identify MPs as potential biomarkers of disease. Moreover, the pathophysiological properties of MPs may contribute to atherosclerosis development. In addition, pharmacological compounds, used in the treatment of cardiovascular disease, can reduce plasma MP concentrations. Nevertheless, numerous issues remain to be solved before MP measurement can be applied as routine biological tests to improve cardiovascular risk prediction. In particular, prospective studies to identify the predictive values of MPs in pathologies such as cardiovascular diseases are needed to demonstrate whether MPs are useful biomarkers for the early detection of the disease and its progression. Quelle: Journal of Cellular and Molecular Medicine | 3 Nov 2011 | 12:46 pm CET

Localized colonic stem cell transplantation enhances tissue regeneration in murine colitis Abstract Many patients suffer from chronic gastrointestinal diseases characterized by chronic inflammation, increased intestinal permeability, and visceral pain in which there is no definitive treatment. Adult stem cells have recently been used in various disease states to contribute wound healing processes. In the current study we investigated the ability of intra-colonic adult stem cells application to heal colonic inflammation in IL-10−/− mice with active colitis. The aims of this study were to determine whether intra-colonic infusion of adult colonic stem cells (local stem cell transplantation): (i) restores intestinal permeability; (ii) attenuates visceral hypersensitivity; (iii) heals murine colitis. IL-10−/− mice with active colitis were transplanted with adult stem cells. Mice received either a single intracolonic infusion of colonic stem cells (CSCs) or colonic epithelial cells (CECs). Two weeks after transplantation, we measured visceral hypersensitivity and intestinal permeability and correlated these with histological improvement of colitis. IL-10−/− mice that received stem cell transplantation showed histopathologic evidence of recovery from colitis. Improvement in colitis as graded by pathology scores correlated with restoration of intestinal permeability and decreased visceral hypersensitivity. Intra-colonic administration of CSCs is a potential therapeutic method for treating refractory symptoms in patients with chronic gastrointestinal diseases associated with chronic inflammation and visceral hypersensitivity. This method may be safer and should have far fewer side effects than systemic stem cell administration. Quelle: Journal of Cellular and Molecular Medicine | 3 Nov 2011 | 12:46 pm CET

Hypoxia-inducible factor 1-mediated human GATA1 induction promotes erythroid differentiation under hypoxic conditions Abstract Hypoxia-inducible factor promotes erythropoiesis through coordinated cell type-specific hypoxia responses. GATA1 is essential to normal erythropoiesis and plays a crucial role in erythroid differentiation. In this study, we show that hypoxia-induced GATA1 expression is mediated by HIF1 in erythroid cells. Under hypoxic conditions, significantly increased GATA1 mRNA and protein levels were detected in K562 cells and erythroid induction cultures of CD34+ hematopoietic stem/progenitor cells. Enforced HIF1α expression increased GATA1 expression, while HIF1α knock-down by RNA interference decreased GATA1 expression. In silico analysis revealed one potential hypoxia response element (HRE). The results from reporter gene and mutation analysis suggested that this element is necessary for hypoxic response. Chromatin immunoprecipitation (ChIP)-PCR showed that the putative HRE was recognized and bound by HIF1 in vivo. These results demonstrate that the up-regulation of GATA1 during hypoxia is directly mediated by HIF1.The mRNA expression of some erythroid differentiation markers was increased under hypoxic conditions, but decreased with RNA interference of HIF1α or GATA1. Flow cytometry analysis also indicated that hypoxia or desferrioxamine or CoCl2 induced expression of erythroid surface marker CD71 and CD235a, while expression repression of HIF1α or GATA1 by RNA interference led to a decreased expression of CD235a. These results suggested that HIF1-mediated GATA1 upregulation promotes erythropoiesis in order to satisfy the needs of an organism under hypoxic conditions. Quelle: Journal of Cellular and Molecular Medicine | 3 Nov 2011 | 12:46 pm CET

Modulation of miroRNA 20b with resveratrol and longevinex is linked with their potent anti-angiogenic action in the ischemic myocardium and synergestic effects of resveratrol and γ-tocotrienol Abstract Background: Resveratrol, a constituent of red wine, and γ-tocotrienol, a constituent of palm oil are important for cardioprotection. Although MicroRNAs are known regulators for genes involved in cardiac remodeling the regulatory pathway involving microRNA has not been studied so far. Methods: We explored the cardioprotection by resveratrol, longevinex and γtocotrienol in ischemia/reperfusion(I/R) model of rat and determined miRNA profile from isolated RNA. Systemic analyses of miRNA array and theirs targets were determined using a number of computational approaches. Results: Resveratrol and γ-tocotrienol, either alone or in combination, modulated the expression pattern of miRNAs close to the control level based on PCA analyses. Differential expression was observed in over 75 miRNAs, some of them, such as miR-21 and miR-20b (antiangiogenic) were previously implicated in cardiac remodeling. The target genes for the highest differentially expressed miRNA include genes of various molecular function such as TGFβ1–Smad3 signaling pathway, inflammation and their transcription factors, which may play key role in reducing I/R injury. Administration of antagomiR-20 attenuated I/R induced VEGF and HIF1α level. Conclusion: All the interventions treated for 3 weeks lead to significant cardioprotection against ischemia/reperfusion injury. A unique signature of miRNA profile is observed in control heart pretreated with resveratrol or γ-tocotrienol. We have determined specific group of miRNA in heart that have altered during IR injuries. Most of those altered microRNA expressions modulated close to their basal level in resveratrol or longevinex treated I/R rat. Interestingly, resveratrol and γ-tocotrienol resulted in synergestic action. Quelle: Journal of Cellular and Molecular Medicine | 3 Nov 2011 | 12:46 pm CET

Inhibition of JAK2/STAT3 signaling induces colorectal cancer cell apoptosis via mitochondrial pathway Abstract Abnormalities in the JAK2/STAT3 pathway are involved in the pathogenesis of colorectal cancer (CRC), including apoptosis. However, the exact mechanism by which dysregulated JAK2/STAT3 signaling contributes to the apoptosis has not been clarified. To investigate the role of both JAK2 and STAT3 in the mechanism underlying CRC apoptosis, we inhibited JAK2 with AG490 and depleted STAT3 with a small interfering RNA (siRNA); Our data showed that inhibition of JAK2/STAT3 signaling induced CRC cellular apoptosis via modulating the Bcl-2 gene family, promoting the loss of mitochondrial transmembrane potential (Δψm) and the increase of reactive oxygen species (ROS). In addition, our results demonstrated that the translocation of cytochrome c (Cyt c), caspase activation and cleavage of poly (ADP-ribose) polymerase (PARP) were present in apoptotic CRC cells after down-regulation of JAK2/STAT3 signaling. Moreover, inhibition of JAK2/STAT3 signaling suppressed CRC xenograft tumor growth. We found that JAK2/STAT3 target genes were decreased meanwhile caspase cascade was activated in xenograft tumors. Our findings illustrated the biological significance of JAK2/STAT3 signaling in CRC apoptosis, and provided novel evidence that inhibition of JAK2/STAT3 induced apoptosis via the mitochondrial apoptotic pathway. Therefore, JAK2/STAT3 signaling may be a potential target for therapy of CRC. Quelle: Journal of Cellular and Molecular Medicine | 2 Nov 2011 | 5:40 pm CET

Epigenetic control of CCR5 transcript levels in immune cells and modulation by small molecules inhibitors Abstract Previously, we have shown that CCR5 transcription is regulated by CREB-1. However, the ubiquitous pattern of CREB-1 expression suggests the involvement of an additional level of transcriptional control in the cell type-specific expression of CCR5. In this study we show that epigenetic changes (i.e. DNA methylation and histone modifications) within the context of the CCR5 P1 promoter region correlate with transcript levels of CCR5 in healthy and in malignant CD4+ T lymphocytes as well as in CD14+ monocytes. In normal naïve T cells and CD14+ monocytes the CCR5 P1 promoter resembles a bivalent chromatin state, with both repressive and permissive histone methylation and acetylation marks. The CCR5 expressing CD14+ monocytes however show much higher levels of acetylated histone H3 (AcH3) compared to the non–CCR5-expressing naïve T cells. Combined with a highly methylated promoter in CD14+ monocytes, this indicates a dominant role for AcH3 in CCR5 transcription. We also show that pharmacological interference in the epigenetic repressive mechanisms that account for the lack of CCR5 transcription in T leukemic cell lines results in an increase in CREB-1 association with the CCR5 P1 chromatin. Furthermore RNA polymerase II was also recruited into CCR5 P1 chromatin resulting in CCR5 re-expression. Together, these data indicate that epigenetic modifications of DNA, and of histones, contribute to the control of CCR5 transcription in immune effector cells. Quelle: Journal of Cellular and Molecular Medicine | 2 Nov 2011 | 5:40 pm CET

The role of dorsal root ganglia activation and brain derived neurotrophic factor in multiple sclerosis Abstract Multiple sclerosis (MS) is characterised by focal destruction of the white matter of the brain and spinal cord. The exact mechanisms underlying the pathophysiology of the disease are unknown. Many studies have shown that MS is predominantly an autoimmune disease with an inflammatory phase followed by a demyelinating phase. Recent studies alongside current treatment strategies, including glatiramer acetate, have revealed a potential role for brain derived neurotrophic factor (BDNF) in MS. However, the exact role of BDNF is not fully understood. We used the experimental autoimmune encephalomyelitis (EAE) model of MS in adolescent female Lewis rats to identify the role of BDNF in disease progression. DRG and spinal cords were harvested for protein and gene expression analysis every 3 days post-disease induction (pdi) up to 15 days. We show significant increases in BDNF protein and gene expression in the DRG of EAE animals at 12 dpi, which correlates with peak neurological disability. BDNF protein expression in the spinal cord was significantly increased at 12 dpi, and maintained at 15 dpi. However, there was no significant change in mRNA levels. We show evidence for the anterograde transport of BDNF protein from the DRG to the dorsal horn of the spinal cord via the dorsal roots. Increased levels of BDNF within the DRG and spinal cord in EAE may facilitate myelin repair and neuroprotection in the CNS. The anterograde transport of DRG derived BDNF to the spinal cord may have potential implications in facilitating central myelin repair and neuroprotection. Quelle: Journal of Cellular and Molecular Medicine | 2 Nov 2011 | 5:39 pm CET

Involvement of COX-2/PGE2 signaling in hypoxia-induced angiogenic response in endothelial cells Abstract Objective: To evaluate the impact of hypoxia on the angiogenic capability of endothelial cells (ECs), and further investigate whether the COX-2/PGE2 signaling is involved in the angiogenic response of ECs to hypoxia. Methods: We explored the impact of various periods (1, 3, 6, 12, 24 h) of hypoxia (2% O2) on human umbilical vein endothelial cells (HUVECs) in vitro. We observed cell viability, migration, tube formation, analyzed COX-2, VEGF, AQP1 mRNA transcription, protein expression and measured PGE2, VEGF protein concentration in cell supernatants. Then we treated HUVECs with COX-2 selective inhibitor NS398, EP1/2 combined antagonist AH6809 and exogenous PGE2 to investigate the role of COX-2/PGE2 signaling in the angiogenic response of ECs to hypoxia. Results: The results demonstrated that short-term hypoxic treatment enhanced HUVECs proliferation, migration, tube formation, significantly up-regulated COX-2, VEGF, AQP1 mRNA level, protein expression and promoted PGE2, VEGF release. The pharmacological inhibition study revealed that exposure of HUVEC to NS398 and AH6809 under hypoxia impaired the biological responses of ECs to hypoxia. Exogenous PGE2 augments the effects of hypoxia on HUVECs, and partially reversed the inhibitory effects of NS398 on HUVECs proliferation and angiogenic capability. Conclusion: Short-term hypoxic treatment enhanced angiogenic capability of ECs, and COX-2/PGE2 signaling maybe play a critical role in the biological response of ECs to hypoxia. Quelle: Journal of Cellular and Molecular Medicine | 2 Nov 2011 | 5:39 pm CET

Pancreatic cancer tumor initiating cells: the molecular regulation and therapeutic values Abstract Pancreatic cancer is an aggressive solid tumor characterized by its local invasion, early metastasis, and resistance to standard chemotherapy or radiation therapy. Tumor initiating cells (TICs) are not only capable of self-renewal and differentiation, but also play an important role in multidrug resistance, and thus become a popular topic in cancer research especially in pancreatic cancer. In this review, we summarize the current progress of TICs in tumorigenesis, various newly identified surface markers of pancreatic TICs, and the signaling pathways such as EMT, SHH and Notch that regulate TICs. We also discuss the role which microRNA plays in TICs as well as its application in TICs-targeted therapy along with other approaches. Quelle: Journal of Cellular and Molecular Medicine | 2 Nov 2011 | 5:38 pm CET

Repetitive transplantation of different cell types sequentially improves heart function after infarction Abstract Cell-based therapy is considered a novel and potentially new strategy in regenerative medicine. But the efficacy of cell-based therapy has been limited by the poor survival of the transplanted cells in an ischemic environment. The goal of the present study is to present a possibility to increase survival of the transplanted cardiomyocytes, by increasing the vascularisation of the infarcted area. First, we injected endothelial progenitor cells (EPC) to augment the vascular density in infarcted areas and to improve the benefit of a subsequent Tx of fetal cardiomyocytes. Serial echocardiography indeed showed significant improvement of the left ventricular function after application of EPC and a significant additive improvement after Tx of fetal cardiomyocytes. In contrast, repetitive EPC transplantation as a control group did not show an additional improvement after the second transplantation. Histological, cells could be readily detected after Tx by BrdU-staining for EPC and by carboxy-fluorescein diacetate succinimidyl ester (CFSE)-staining for fetal cardiomyocytes. Staining for CD31 revealed a significant increase in vessel density in the infarction area compared with medium controls, possibly contributing to the benefit of transplanted foetal cardiomyocytes. Notably, a significant increase in the number of apoptotic cells was observed in cell-transplanted hearts was accompanied by an increase in proliferation, collagen content and neutrophil infiltration, suggesting an active remodelling concomitant with sustained inflammatory processes. In conclusion, repetitive Tx of different cell types after myocardial infarction in rat hearts significantly improved left ventricular function and could represent a feasible option to enhance the benefit of cell therapy. Quelle: Journal of Cellular and Molecular Medicine | 2 Nov 2011 | 5:38 pm CET

Cardiomyocytes generated from CPVTD307H patients are arrhythmogenic in response to β-adrenergic stimulation Abstract Sudden cardiac death caused by ventricular arrhythmias is a disastrous event, especially when it occurs in young individuals. Among the 5 major arrhythmogenic disorders occurring in the absence of a structural heart disease is Catecholaminergic Polymorphic Ventricular Tachycardia (CPVT), which is a highly lethal form of inherited arrhythmias. Our study focuses on the autosomal recessive form of the disease caused by the missense mutation D307H in the cardiac calsequestrin gene, CASQ2. Since CASQ2 is a key player in excitation contraction coupling, the derangements in intracellular Ca2+ handling may cause delayed afterdepolarizations (DADs) which constitute the mechanism underlying CPVT. To investigate catecholamine-induced arrhythmias in the CASQ2 mutated cells, we generated for the first time CPVT-derived iPSCs by reprogramming fibroblasts from skin biopsies of two patients, and demonstrated that the iPSCs carry the CASQ2 mutation. Next, iPSCs were differentiated to cardiomyocytes (iPSCs-CMs) which expressed the mutant CASQ2 protein. The major findings were that the β-adrenergic agonist isoproterenol caused in CPVT iPSCs-CMs (but not in the control cardiomyocytes) DADs, oscillatory arrhythmic prepotentials, after-contractions and diastolic [Ca2+]i rise. Electron microscopy analysis revealed that compared with control iPSCs-CMs, CPVT iPSCs-CMs displayed a more immature phenotype with less organized myofibrils, enlarged sarcoplasmic reticulum cisternae and reduced number of caveolae. In summary, our results demonstrate that the patient-specific mutated cardiomyocytes can be used to study the electrophysiological mechanisms underlying CPVT. Quelle: Journal of Cellular and Molecular Medicine | 2 Nov 2011 | 5:38 pm CET

Activation of neurokinin-1 receptors up-regulates substance P and neurokinin-1 receptor expression in murine pancreatic acinar cells Abstract Acute pancreatitis (AP) has been associated with an up-regulation of substance P (SP) and neurokinin-1 receptor (NK1R) in the pancreas. Increased SP-NK1R interaction was suggested to be pro-inflammatory during AP. Previously, we showed that caerulein treatment increased SP/NK1R expression in mouse pancreatic acinar cells, but the effect of SP treatment was not evaluated. Pancreatic acinar cells were obtained from pancreas of male swiss mice (25–30g). We measured mRNA expression of preprotachykinin-A (PPTA) and NK1R following treatment of SP (10−6M). SP treatment increased PPTA and NK1R expression in isolated pancreatic acinar cells, which was abolished by pretreatment of a selective NK1R antagonist, CP96,345. SP also time dependently increased protein expression of NK1R. Treatment of cells with a specific NK1R agonist, GR73,632, up-regulated SP protein levels in the cells. Using previously established concentrations, pre-treatment of pancreatic acinar cells with Gö6976 (10nM), rottlerin (5μM), PD98059 (30μM), SP600125 (30μM) or Bay11–7082 (30μM) significantly inhibited up-regulation of SP and NK1R. These observations suggested that the PKC-ERK/JNK-NF-κB pathway is necessary for the modulation of expression levels. In comparison, pre-treatment of CP96,345 reversed gene expression in SP-induced cells, but not in caerulein-treated cells. Overall, the findings in this study suggested a possible auto-regulatory mechanism of SP/NK1R expression in mouse pancreatic acinar cells, via activation of NK1R. Elevated SP levels during AP might increase the occurrence of a positive feedback loop that contributes to abnormally high expression of SP and NK1R. Quelle: Journal of Cellular and Molecular Medicine | 1 Nov 2011 | 6:39 am CET

SIRT1 is required for long-term growth of human mesenchymal stem cells Abstract   Human mesenchymal stem cells (MSCs) have therapeutic potential because of their ability to self-renew and differentiate into multiple tissues. However, senescence often occurs in MSCs when they are cultured in vitro and the molecular mechanisms underlying this effect remain unclear. In this study, we found that NAD-dependent protein deacetylase SIRT1 is differentially expressed in both human bone marrow-derived MSCs (B-MSCs) and adipose tissue-derived MSCs after increasing passages of cell culture. Using lentiviral shRNA we demonstrated that selective knockdown of SIRT1 in human MSCs at early passage slows down cell growth and accelerates cellular senescence. Conversely, overexpression of SIRT1 delays senescence in B-MSCs that have undergone prolonged in vitro culturing and the cells do not lose adipogenic and osteogenic potential. In addition, we found that the delayed accumulation of the protein p16 is involved in the effect of SIRT1. However, resveratrol, which has been used as an activator of SIRT1 deacetylase activity, only transiently promotes proliferation of B-MSCs. Our findings will help us understand the role of SIRT1 in the aging of normal diploid cells and may contribute to the prevention of human MSCs senescence thus benefiting MSCs-based tissue engineering and therapies. Content Type Journal Article Category Original Article Pages 1-12 DOI 10.1007/s00109-011-0825-4 Authors Hong-Feng Yuan, Stem Cell and Regeneration Medicine Lab, Beijing Institute of Transfusion Medicine, 27, Taiping Road, Beijing, 100850 China Chao Zhai, Stem Cell and Regeneration Medicine Lab, Beijing Institute of Transfusion Medicine, 27, Taiping Road, Beijing, 100850 China Xin-Long Yan, Stem Cell and Regeneration Medicine Lab, Beijing Institute of Transfusion Medicine, 27, Taiping Road, Beijing, 100850 China Dan-Dan Zhao, Department of Hematology, The First Affiliated Hospital of PLA General Hospital, 51, Fucheng Road, Beijing, China 100037 Jing-Xue Wang, Stem Cell and Regeneration Medicine Lab, Beijing Institute of Transfusion Medicine, 27, Taiping Road, Beijing, 100850 China Quan Zeng, Stem Cell and Regeneration Medicine Lab, Beijing Institute of Transfusion Medicine, 27, Taiping Road, Beijing, 100850 China Lin Chen, Stem Cell and Regeneration Medicine Lab, Beijing Institute of Transfusion Medicine, 27, Taiping Road, Beijing, 100850 China Xue Nan, Stem Cell and Regeneration Medicine Lab, Beijing Institute of Transfusion Medicine, 27, Taiping Road, Beijing, 100850 China Li-Juan He, Stem Cell and Regeneration Medicine Lab, Beijing Institute of Transfusion Medicine, 27, Taiping Road, Beijing, 100850 China Si-Ting Li, Stem Cell and Regeneration Medicine Lab, Beijing Institute of Transfusion Medicine, 27, Taiping Road, Beijing, 100850 China Wen Yue, Stem Cell and Regeneration Medicine Lab, Beijing Institute of Transfusion Medicine, 27, Taiping Road, Beijing, 100850 China Xue-Tao Pei, Stem Cell and Regeneration Medicine Lab, Beijing Institute of Transfusion Medicine, 27, Taiping Road, Beijing, 100850 China Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 29 Oct 2011 | 7:43 am CEST

Adenosine A2A receptor activation stimulates collagen production in sclerodermic dermal fibroblasts either directly and through a cross-talk with the cannabinoid system Abstract   Systemic sclerosis (SSc) is a connective tissue disease characterised by exaggerated collagen deposition in the skin and visceral organs. Adenosine A2A receptor stimulation (A2Ar) promotes dermal fibrosis, while the cannabinoid system modulates fibrogenesis in vitro and in animal models of SSc. Moreover, evidence in central nervous system suggests that A2A and cannabinoid (CB1) receptors may physically and functionally interact. On this basis, we investigated A2Ar expression and function in modulating collagen biosynthesis from SSc dermal fibroblasts and analysed the cross-talk with cannabinoid receptors. In sclerodermic cells, A2Ar expression (RT-PCR, Western blotting) was evaluated together with the effects of A2A agonists and/or antagonists on collagen biosynthesis (EIA, Western blotting). Putative physical and functional interactions between the A2A and cannabinoid receptors were respectively assessed by co-immuno-precipitation and co-incubating the cells with the unselective cannabinoid agonist WIN55,212-2, and the selective A2A antagonist ZM-241385. In SSc fibroblasts, (1) the A2Ar is overexpressed and its occupancy with the selective agonist CGS-21680 increases collagen production, myofibroblast trans-differentiation, and ERK-1/2 phosphorylation; (2) the A2Ar forms an heteromer with the cannabinoid CB1 receptor; and (3) unselective cannabinoid receptor stimulation with a per se ineffective dose of WIN55,212-2, results in a marked anti-fibrotic effect after A2Ar blockage. In conclusion, A2Ar stimulation induces a pro-fibrotic phenotype in SSc dermal fibroblasts, either directly, and indirectly, by activating the CB1 cannabinoid receptor. These findings increase our knowledge of the pathophysiology of sclerodermic fibrosis also further suggesting a new therapeutic approach to the disease. Content Type Journal Article Category Original Article Pages 1-12 DOI 10.1007/s00109-011-0824-5 Authors Pietro Enea Lazzerini, Department of Clinical Medicine and Immunological Sciences, Division of Clinical Immunology, University of Siena, Siena, Italy Mariarita Natale, Department of Clinical Medicine and Immunological Sciences, Division of Clinical Immunology, University of Siena, Siena, Italy Elena Gianchecchi, Department of Clinical Medicine and Immunological Sciences, Division of Clinical Immunology, University of Siena, Siena, Italy Pier Leopoldo Capecchi, Department of Clinical Medicine and Immunological Sciences, Division of Clinical Immunology, University of Siena, Siena, Italy Cinzia Montilli, Department of Clinical Medicine and Immunological Sciences, Division of Clinical Immunology, University of Siena, Siena, Italy Stefania Zimbone, Department of Clinical Medicine and Immunological Sciences, Division of Clinical Immunology, University of Siena, Siena, Italy Monica Castrichini, Department of Clinical Medicine and Immunological Sciences, Division of Clinical Immunology, University of Siena, Siena, Italy Epifania Balistreri, Department of Clinical Medicine and Immunological Sciences, Division of Rheumatology, University of Siena, Siena, Italy Gianluca Ricci, Department of Clinical Medicine and Immunological Sciences, Division of Clinical Immunology, University of Siena, Siena, Italy Enrico Selvi, Department of Clinical Medicine and Immunological Sciences, Division of Rheumatology, University of Siena, Siena, Italy Estrella Garcia-Gonzalez, Department of Clinical Medicine and Immunological Sciences, Division of Rheumatology, University of Siena, Siena, Italy Mauro Galeazzi, Department of Clinical Medicine and Immunological Sciences, Division of Rheumatology, University of Siena, Siena, Italy Franco Laghi-Pasini, Department of Clinical Medicine and Immunological Sciences, Division of Clinical Immunology, University of Siena, Siena, Italy Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 28 Oct 2011 | 8:04 am CEST

An intronic MYLK variant associated with inflammatory lung disease regulates promoter activity of the smooth muscle myosin light chain kinase isoform Abstract   Intronic single-nucleotide polymorphisms (SNPs) are commonly associated with complex diseases but exhibit unknown biologic functionality. Myosin light-chain kinase (MLCK), a central cytoskeletal regulator encoded by MYLK, plays a key pathophysiological role in complex diseases including acute lung injury (ALI) and asthma. We studied the potential regulatory roles of two intronic MYLK SNPs (rs936170 and rs820336) previously associated with ALI and asthma. Due to their genomic location at the junction encoding the non-muscle and smooth muscle MLCK (smMLCK) isoforms, we first identified the transcription start site (TSS) of the smMLCK isoform, and isolated a 2,954-bp DNA fragment upstream of the smMLCK TSS. Serial 5′ deletion of the fragment revealed a proximal promoter region exhibiting strong promoter activity with potential inhibitory elements in the distal region. Site-directed mutageneses and luciferase reporter assays showed no effect of the distal promoter SNP rs936170 on smMLCK promoter activity. In contrast, SNP rs820336, located in an enhancer/repressor region downstream of TSS, was identified to regulate smMLCK promoter activity in an allelic-dependent manner. The A allele interrupted the binding site for Forkhead box protein N1 (FOXN1), a transcription factor governing expression of immune response genes. Silencing of FOXN1 expression (siRNA) reduced FOXN1 interaction with cis-regulatory elements in proximity to rs820336 and significantly decreased smMLCK expression. These functional insights into the involvement of intronic MYLK SNPs further strengthen the concept that MYLK contributes to inflammatory disease susceptibility and represents an attractive molecular target in complex inflammatory disorders. Content Type Journal Article Category Original Article Pages 1-10 DOI 10.1007/s00109-011-0820-9 Authors Yoo Jeong Han, Section of Pulmonary and Critical Care, Department of Medicine, University of Chicago, Chicago, IL 60637, USA Shwu-Fan Ma, Section of Pulmonary and Critical Care, Department of Medicine, University of Chicago, Chicago, IL 60637, USA Michael S. Wade, Institute for Personalized Respiratory Medicine, Section of Pulmonary Critical Care, Allergy and Sleep Medicine, University of Illinois at Chicago, Chicago, IL 60612, USA Carlos Flores, CIBER de Enfermedades Respiratorias, Instituto de Salud Carlos III, Madrid, Spain Joe G. N. Garcia, Institute for Personalized Respiratory Medicine, Department of Medicine, Pharmacology and Bioengineering, University of Illinois at Chicago, 1737 West Polk Street, 305D AOB, Chicago, IL 60612-7227, USA Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 20 Oct 2011 | 5:47 pm CEST

Glucocorticoid receptor antagonist sensitizes TRAIL-induced apoptosis in renal carcinoma cells through up-regulation of DR5 and down-regulation of c-FLIP(L) and Bcl-2 Abstract   RU486 (Mifepristone) has been known as antiprogesterone and antiglucocorticoid agent. RU486 is also used for treatment of several cancers, such as breast, ovarian, prostate, and glaucoma. Here, we investigated the effect of RU486 on TRAIL-induced apoptosis in human renal carcinoma Caki cells. Low dose of RU486 (30–50 μM) alone had no effect on apoptosis, but RU486 markedly sensitized Caki cells to TRAIL-induced apoptosis. We found that up-regulation of death receptor 5 (DR5; receptor for TRAIL ligand), and down-regulation of Bcl-2 and c-FLIP (caspase regulator) contributes to RU-486 induced TRAIL sensitization. Down-regulation of DR5 by siRNA also blocked RU486 induced TRAIL sensitization. Furthermore, overexpression of Bcl-1 or c-FLIP(L) inhibited the cell death induced by the combined treatment with RU486 and TRAIL. RU486 increased DR5 expression at the transcriptional levels through induction of CHOP expression. By contrast, RU486 did not sensitize normal human mesangial cells to TRAIL-mediated apoptosis. Effect of RU486 on TRAIL-induced cancer cell apoptosis was independent of glucocorticoid receptor and progesterone receptor. Taken together, RU486 enhances TRAIL-mediated apoptosis through down-regulation of Bcl-2 and c-FLIP(L) as well as CHOP-mediated DR5 up-regulation. Content Type Journal Article Category Original Article Pages 1-11 DOI 10.1007/s00109-011-0821-8 Authors Kyoung-jin Min, Department of Immunology, School of Medicine, Keimyung University, 2800 Dalgubeoldaero, Dalseo-Gu, Daegu, 704-701 South Korea Ji Hoon Jang, Department of Immunology, School of Medicine, Keimyung University, 2800 Dalgubeoldaero, Dalseo-Gu, Daegu, 704-701 South Korea Jung Tae Lee, Department of Immunology, School of Medicine, Keimyung University, 2800 Dalgubeoldaero, Dalseo-Gu, Daegu, 704-701 South Korea Kyeong Sook Choi, Department of Molecular Science and Technology, Institute for Medical Sciences, Ajou University School of Medicine, 5 Woncheon-Dong, Paldal-Gu, Suwon, 442-749 South Korea Taeg Kyu Kwon, Department of Immunology, School of Medicine, Keimyung University, 2800 Dalgubeoldaero, Dalseo-Gu, Daegu, 704-701 South Korea Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 19 Oct 2011 | 7:54 am CEST

Heat shock protein 27 as a prognostic and predictive biomarker in pancreatic ductal adenocarcinoma Abstract A role of heat shock protein 27 (HSP27) as a potential biomarker has been reported in various tumor entities, but comprehensive studies in pancreatic cancer are lacking. Applying tissue microarray (TMA) analysis, we correlated HSP27 protein expression status with clinicopathologic parameters in pancreatic ductal adenocarcinoma specimens from 86 patients. Complementary, we established HSP27 overexpression and RNA-interference models to assess the impact of HSP27 on chemo- and radiosensitivity directly in pancreatic cancer cells. In the TMA study, HSP27 expression was found in 49% of tumor samples. Applying univariate analyses, a significant correlation was found between HSP27 expression and survival. In the multivariate Cox-regression model, HSP27 expression emerged as an independent prognostic factor. HSP27 expression also correlated inversely with nuclear p53 accumulation, indicating either protein interactions between HSP27 and p53 or TP53 mutation-dependent HSP27-regulation in pancreatic cancer. In the sensitivity studies, HSP27 overexpression rendered HSP27 low-expressing PL5 pancreatic cancer cells more susceptible towards treatment with gemcitabine. Vice versa, HSP27 protein depletion in HSP27 high-expressing AsPC-1 cells caused increased gemcitabine resistance. Importantly, HSP27 expression was inducible in pancreatic cancer cell lines as well as primary cells. Taken together, our study suggests a role for HSP27 as a prognostic and predictive marker in pancreatic cancer. Assessment of HSP27 expression could thus facilitate the identification of specific patient subpopulations that might benefit from individualized treatment options. Additional studies need to clarify whether modulation of HSP27 expression could represent an attractive concept to support the incorporation of hyperthermia in clinical treatment protocols for pancreatic cancer. Quelle: Journal of Cellular and Molecular Medicine | 18 Oct 2011 | 9:27 am CEST

A novel regulation of VEGF expression by HIF-1α and STAT3 in HDM2 transfected prostate cancer cells Abstract On the basis of increasing roles for HDM2 oncoprotein in cancer growth and progression, we speculated that HDM2 might play a major role in hypoxia-induced metastatic process. For verification of this hypothesis, wild type LNCaP prostate cancer cells and HDM2 transfected LNCaP-MST (HDM2 stably transfected) cells were studied. The data obtained revealed that the HDM2 transfected LNCaP-MST cells possessed the ability to multiply rapidly and show distinct morphological features compared to non-transfected LNCaP cells. During hypoxia HDM2 expression in the LNCaP and LNCaP-MST cells were significantly increased. The LNCaP-MST cells also expressed higher levels of HIF-1α (hypoxia inducible factor-1α) and p-STAT3 even under the normoxic conditions compared to the non-transfected cells. The HIF-1α and p-STAT3 expressions were increased several fold when the cells were subjected to hypoxic conditions. The HIF-1α and p-STAT3 protein expressions observed in HDM2 transfected LNCaP-MST cells were 20 and 15 folds higher respectively, compared to the non-transfected wild type LNCaP cells. These results demonstrate that HDM2 may have an important regulatory role in mediating the HIF-1α and p-STAT3 protein expression during both normoxic and hypoxic conditions. Furthermore, the VEGF expression that is typically regulated by HIF-1α and p-STAT3 were also increased significantly by 136% (P<0.01) after HDM2 transfection. The overall results point towards a novel ability of HDM2 in regulating HIF-1α and p-STAT3 levels even in normoxic conditions that eventually lead to an up-regulation of VEGF expression. Quelle: Journal of Cellular and Molecular Medicine | 18 Oct 2011 | 9:27 am CEST

Bone marrow mesenchymal stem cells for post-myocardial infarction cardiac repair: micro-RNAs as novel regulators Abstract Transplantation of bone marrow-derived mesenchymal stem cells (MSCs) is safe and may improve cardiac function and structural remodeling in patients following myocardial infarction (MI). Cardiovascular cell differentiation and paracrine effects to promote endogenous cardiac regeneration, neovascularization, anti-inflammation, anti-apoptosis, anti-remodeling and cardiac contractility, may contribute to MSC-based cardiac repair following MI. However, current evidence indicates that the efficacy of MSC transplantation was unsatisfactory, due to the poor viability and massive death of the engrafted MSCs in the infarcted myocardium. MicroRNAs are short endogenous, conserved, non-coding RNAs and important regulators involved in numerous facets of cardiac pathophysiologic processes. There is an obvious involvement of microRNAs in almost every facet of putative repair mechanisms of MSC-based therapy in MI, such as stem cell differentiation, neovascularization, apoptosis, cardiac remodeling, cardiac contractility and arrhythmias, and others. It is proposed that therapeutic modulation of individual cardiovascular microRNA of MSCs, either mimicking or antagonizing microRNA actions, will hopefully enhance MSC therapeutic efficacy. In addition, MSCs may be manipulated to enhance functional microRNA expression or to inhibit aberrant microRNA levels in a paracrine manner. We hypothesize that microRNAs may be used as novel regulators in MSC-based therapy in MI and MSC transplantation by microRNA regulation may represent promising therapeutic strategy for MI patients in the future. Quelle: Journal of Cellular and Molecular Medicine | 18 Oct 2011 | 9:26 am CEST

Therapeutic potential of in utero mesenchymal stem cell (MSCs) transplantation in rat fetuses with spina bifida aperta Abstract Neural tube defects (NTDs) are complex congenital malformations resulting from incomplete neurulation in embryo. Despite surgical repair of the defect, most of the patients who survive with NTDs have a multiple system handicap due to neuron deficiency of the defective spinal cord. In this study, we successfully devised a prenatal surgical approach and transplanted mesenchymal stem cells (MSCs) to fetal rat spinal column to treat retinoic acid induced NTDs in rat. Transplanted MSCs survived, grew and expressed markers of neurons, glia and myoblasts in the defective spinal cord. MSCs expressed and perhaps induced the surrounding spinal tissue to express neurotrophic factors. In addition, MSC reduced spinal tissue apoptosis in NTD. Our results suggested that prenatal MSC transplantation could treat spinal neuron deficiency in NTDs by the regeneration of neurons and reduced spinal neuron death in the defective spinal cord. Quelle: Journal of Cellular and Molecular Medicine | 18 Oct 2011 | 9:25 am CEST

Effects of redox modulation by inhibition of Thioredoxin reductase on radiosensitivity and gene expression Abstract The thioredoxin system is a promising target when aiming to overcome the problem of clinical radiation resistance. Altered cellular redox-status and redox sensitive thiols contributing to induction of resistance strongly connects the ubiquitous redox enzyme thioredoxin reductase to the cellular response to ionizing radiation. To further investigate possible strategies in combating clinical radiation resistance, human radio-resistant lung cancer cells were subjected to a combination of single fractions of γ-radiation at clinically relevant doses and non-toxic levels of a well-characterized thioredoxin reductase (TrxR) inhibitor, the phosphine gold(I) compound [Au(SCN)(PEt3)]. The combination of the TrxR-inhibitor and ionizing radiation reduced the surviving fractions and impaired the ability of the U1810 cells to repopulate by approximately 50%. In addition, inhibition of thioredoxin reductase caused changes in the cell cycle distribution, suggesting a disturbance of the mitotic process. Global gene expression analysis also revealed clustered genetic expression changes connected to several major cellular pathways such as cell cycle, cellular response to stress and DNA-damage. Specific TrxR-inhibition as a factor behind the achieved results was confirmed by correlation of gene expression patterns between gold and siRNA treatment. These results clearly demonstrates TrxR as an important factor conferring resistance to irradiation and the use of [Au(SCN)(PEt3)] as a promising radiosensitizing agent. Quelle: Journal of Cellular and Molecular Medicine | 18 Oct 2011 | 9:25 am CEST

Cardiomyogenic differentiation-independent improvement of cardiac function by human cardiomyocyte progenitor cell injection in ischemic mouse hearts Abstract Introduction: We previously showed that human cardiomyocyte progenitor cells (hCMPCs) injected after myocardial infarction (MI) had differentiated into cardiomyocytes in vivo 3 months post MI. Here, we investigated the short-term (2 weeks) effects of hCMPCs on the infarcted mouse myocardium. Methods: MI was induced in immunocompromised (NOD/scid) mice, immediately followed by intramyocardial injection of hCMPCs labeled with enhanced green fluorescent protein (hCMPC group) or vehicle only (control group). Sham-operated mice served as reference. Cardiac performance was measured 2 and 14 days after MI by magnetic resonance imaging at 9.4T. Left ventricular (LV) pressure-volume measurements were performed at day 15 followed by extensive immunohistological analysis. Results: Animals injected with hCMPCs demonstrated a higher LV ejection fraction, lower LV end-systolic volume and smaller relaxation time constant than control animals 14 days post MI. hCMPCs engrafted in the infarcted myocardium, did not differentiate into cardiomyocytes, but increased vascular density and proliferation rate in the infarcted and border zone area of the hCMPC group. Conclusions: Injected hCMPCs engraft into murine infarcted myocardium where they improve LV systolic function and attenuate the ventricular remodeling process 2 weeks post MI. Since no cardiac differentiation of hCMPCs was evident after 2 weeks, the observed beneficial effects were most likely mediated by paracrine factors, targeting amongst others vascular homeostasis. These results demonstrate that hCMPCs can be applied to repair infarcted myocardium without the need to undergo differentiation into cardiomyocytes. Quelle: Journal of Cellular and Molecular Medicine | 18 Oct 2011 | 9:24 am CEST

MicroRNA expression in formalin-fixed paraffin embedded tissue using Real Time Quantitative PCR; the strengths and pitfalls Abstract MicroRNAs (miRNAs) are a group of small non-coding RNAs with a huge impact in a wide range of biological processes, including cancer. The evidence collected to date demonstrates that miRNAs represent valid diagnostic, prognostic, and predictive markers in cancer. The identification of these miRNA biomarkers in archived tissues has been facilitated by novel development and refinement of detection methodologies. Quantitative real-time reverse-transcription PCR (qRT-PCR) is one of the most common methods used to detect low levels of miRNAs with high sensitivity and specificity. However, several technical parameters should be identified and optimized in order to obtain meaningful and reproducible results. The purpose of this review is to describe some of these technical parameters and improve the validity and reliability of miRNA expression studies. Quelle: Journal of Cellular and Molecular Medicine | 18 Oct 2011 | 9:24 am CEST

The new low-toxic histone deacetylase inhibitor S-(2) induces apoptosis in various acute myeloid leukemia cells Abstract Histone deacetylase inhibitors (HDACi) induce tumor cell cycle arrest and/or apoptosis, and some of them are currently used in cancer therapy. Recently, we described a series of powerful HDACi characterized by a 1,4-benzodiazepine (BDZ) ring hybridized with a linear alkyl chain bearing a hydroxamate function as Zn++-chelating group. Here, we explored the anti-leukemic properties of three novel hybrids, namely the chiral compounds (S)-2 and (R)-2, and their non-chiral analog 4, which were first comparatively tested in promyelocytic NB4 cells. (S)-2 and partially 4– but not (R)-2 – caused G0/G1 cell-cycle arrest by up-regulating cyclin G2 and p21 expression and down-regulating cyclin D2 expression, and also apoptosis as assessed by cell morphology and cytofluorimetric assay, histone H2AX phosphorylation and PARP cleavage. Notably, these events were partly prevented by an anti-oxidant. Moreover, novel HDACi prompted p53 and α-tubulin acetylation and, consistently, inhibited HDAC1 and 6 activity. The rank order of potency was (S)-2 > 4 > (R)-2, reflecting that of other biological assays and addressing (S)-2 as the most effective compound capable of triggering apoptosis in various acute myeloid leukemia (AML) cell lines and blasts from patients with different AML subtypes. Importantly, (S)-2 was safe in mice (up to 150 mg/kg/week) as determined by liver, spleen, kidney and bone marrow histopathology; and displayed negligible affinity for peripheral/central BDZ-receptors. Overall, the BDZ-hydroxamate (S)-2 showed to be a low-toxic HDACi with powerful anti-proliferative and pro-apototic activities towards different cultured and primary AML cells, and therefore of clinical interest to support conventional anti-leukemic therapy. Quelle: Journal of Cellular and Molecular Medicine | 17 Oct 2011 | 2:23 pm CEST

The plaque protein myozap identified as a novel major component of adhering junctions in endothelia of the blood and the lymph vascular systems Abstract Recently the protein myozap (myocardium-enriched zonula adhaerens protein), a 54-kDa polypeptide, which is not a member of any of the known cytoskeletal and junctional protein multigene families, has been identified as a constituent of the plaques of the composite junctions in the intercalated disks connecting the cardiomyocytes of mammalian hearts. Using a set of novel, highly sensitive and specific antibodies we now report that myozap is also a major constituent of the cytoplasmic plaques of the adherens junctions (AJs) connecting the endothelial cells of the mammalian blood and lymph vascular systems, including the desmoplakin-containing complexus adhaerentes of the virgultar cells of lymph node sinus. In light and electron microscopic immunolocalization experiments we show that myozap colocalizes with several proteins of desmosomal plaques as well as with AJ-specific transmembrane molecules, including VE-cadherin. In biochemical analyses, rigorous immunoprecipitation experiments have revealed N-cadherin, desmoplakin, desmoglein-2, plakophilin-2, plakoglobin and plectin as very stably bound complex partners. We conclude that myozap is a general component of cell-cell junctions not only in the myocardium but also in diverse endothelia of the blood and lymph vascular systems of adult mammals, suggesting that this protein not only serves a specific role in the heart but also a broader set of functions in the vessel systems. We also propose to use myozap as an endothelial cell type marker in diagnoses. Quelle: Journal of Cellular and Molecular Medicine | 13 Oct 2011 | 6:20 am CEST

Implication of CD38 gene in podocyte epithelial-to-mesenchymal transition and glomerular sclerosis Abstract CD38 is a multifunctional protein involving in a number of signaling pathways. Given that the lack of CD38 is considered as a dedifferentiation marker of lymphocytes and other cells, we hypothesized that CD38 and its signaling pathway may participate in the epithelial-to-mesenchymal transition (EMT) process of podocytes and thereby regulates the integrity of glomerular structure and function. Western blot analysis and RT-PCR demonstrated that renal tissue CD38 expression was lacking in CD38−/− mice or substantially reduced in renal CD38 shRNA-transfected WT (CD38-shRNA) mice compared to CD38+/+ littermates. Confocal fluorescent microscopy demonstrated the reduced expression of epithelial markers (P-Cadherin, ZO-1 and podocin) and increased expression of mesenchymal markers (FSP-1, α-SMA and desmin) in the glomeruli of CD38−/− and CD38-shRNA mice compared to CD38+/+ mice. Morphological examinations showed profound injury in the glomeruli of CD38−/− or CD38-shRNA mice compared to CD38+/+ mice. This enhanced glomerular injury in CD38−/− or CD38-shRNA mice was accompanied by increased albuminuria and proteinuria. DOCA/high salt treatment further decreased the expression of epithelial markers and increased the abundance of mesenchymal markers, which were accompanied by more increased glomerular damage index and mean arterial pressure in CD38−/- and CD38-shRNA mice than CD38+/+ mice. In vitro studies showed that inhibition of CD38 enhances the EMT in podocytes. In conclusion, our observations reveal that the normal expression of CD38 importantly contributes to the differentiation and function of podocytes and the defect of this gene expression may be a critical mechanism inducing EMT and consequently resulting in glomerular injury and sclerosis. Quelle: Journal of Cellular and Molecular Medicine | 13 Oct 2011 | 6:20 am CEST

Rv0315, a novel immunostimulatory antigen of Mycobacterium tuberculosis, activates dendritic cells and drives Th1 immune responses Abstract   Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) is one of the most deadly infectious diseases, with approximately two million people dying of TB annually. An effective therapeutic method for activating dendritic cells (DCs) and driving Th1 immune responses would improve host defenses and further the development of a TB vaccine. Given the importance of DC maturation in eliciting protective immunity against TB, we investigated whether Rv0315, a newly identified Mtb antigen, can prompt DC maturation. We found that Rv0315 functionally activated DCs by augmenting the expression of the co-stimulatory molecules CD80 and CD86 as well as MHC class I/II molecules. Moreover, it increased DC secretion of the pro-inflammatory cytokines IL-6, IL-1β, and TNF-α. Unlike LPS, however, Rv0315 induced the secretion of IL-12p70, but not IL-10. In addition, Rv0315-treated DCs accelerated the proliferation of CD4+ and CD8+ splenic T cells from Mtb-infected mice, with increased levels of IFN-γ, in syngeneic and allogeneic mixed lymphocyte reactions, indicating that Rv0315 contributes to Th1 polarization of the immune response. Importantly, both mitogen-activated protein kinases and nuclear factor κB signaling mediated the expression of DC surface markers and cytokines. Taken together, our results indicate that Rv0315 is a novel DC maturation-inducing antigen that drives T cell immune responses toward Th1 polarization, suggesting that Rv0315 plays a key role in determining the nature of the immune response to TB. Content Type Journal Article Category Original Article Pages 1-14 DOI 10.1007/s00109-011-0819-2 Authors Eui-Hong Byun, Department of Microbiology and Research Institute for Medical Sciences, College of Medicine, Chungnam National University, Daejeon, 301-747 South Korea Woo Sik Kim, Department of Microbiology and Research Institute for Medical Sciences, College of Medicine, Chungnam National University, Daejeon, 301-747 South Korea A-Rum Shin, Department of Microbiology and Research Institute for Medical Sciences, College of Medicine, Chungnam National University, Daejeon, 301-747 South Korea Jong-Seok Kim, Department of Microbiology and Research Institute for Medical Sciences, College of Medicine, Chungnam National University, Daejeon, 301-747 South Korea Jake Whang, Department of Microbiology and Research Institute for Medical Sciences, College of Medicine, Chungnam National University, Daejeon, 301-747 South Korea Choul-Jae Won, Department of Microbiology and Research Institute for Medical Sciences, College of Medicine, Chungnam National University, Daejeon, 301-747 South Korea Yohan Choi, Department of Microbiology and Research Institute for Medical Sciences, College of Medicine, Chungnam National University, Daejeon, 301-747 South Korea Su-Young Kim, Division of Pulmonary and Critical Care Medicine, Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, 135-710 South Korea Won Jung Koh, Division of Pulmonary and Critical Care Medicine, Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, 135-710 South Korea Hwa-Jung Kim, Department of Microbiology and Research Institute for Medical Sciences, College of Medicine, Chungnam National University, Daejeon, 301-747 South Korea Sung Jae Shin, Department of Microbiology and Research Institute for Medical Sciences, College of Medicine, Chungnam National University, Daejeon, 301-747 South Korea Journal Journal of Molecular Medicine Online ISSN 1432-1440 Print ISSN 0946-2716 Quelle: Journal of Molecular Medicine (Online First™) | 12 Oct 2011 | 6:32 pm CEST

Human umbilical cord mesenchymal stem cells suppress breast cancer tumorigenesis through direct cell-cell contact and internalization Abstract The purpose of this study was to investigate how HUMSCs affect breast cancer tumorigenesis. To observe the influence of HUMSCs on tumorigenesis in vitro, we performed a co-culture of MDA MB-231 breast cancer cells with HUMSCs, and a result of HUMSCs on tumorigenesis in vivo was achieved by injection of HUMSCs into NOD/SCID mice following tumor establishment with MDA-MB231. During the co-culture, apoptosis of MDA-MB231 was noted, which was driven either by binding with HUMSC through direct cell-cell contact or by formation of a novel cell-in-cell phenomenon after internalization of HUMSC. Also, treatment with HUMSC injection was efficacious in both in situ and metastatic breast cancers in the animal models. Since HUMSCs were proved to efficaciously suppress breast cancer tumorigenesis both in vitro and in vivo, it is our expectation that treatment with HUMSCs can be a viable therapy for breast cancer in the near future. In addition, we share a new point of view on the role of HUMSCs in fetal development during pregnancy. Quelle: Journal of Cellular and Molecular Medicine | 5 Oct 2011 | 8:05 am CEST

Duodenal expression of iron transport molecules in patients with hereditary hemochromatosis or iron deficiency Abstract Disturbances of iron metabolism are observed in chronic liver diseases. In the present study, we examined gene expression of duodenal iron transport molecules and hepcidin in patients with hereditary hemochromatosis (HHC) (treated and untreated), involving various genotypes (genotypes which represent risk for HHC were examined), and in patients with iron deficiency anemia (IDA). Gene expressions of DMT1, ferroportin, Dcytb, hephaestin, HFE, and TFR1 were measured in duodenal biopsies using real-time PCR and Western blot. Serum hepcidin levels were measured using ELISA. DMT1, ferroportin, and TFR1 mRNA levels were significantly increased in post-phlebotomized hemochromatics relative to controls. mRNAs of all tested molecules were significantly increased in patients with IDA compared to controls. The protein expression of ferroportin was increased in both groups of patients but not significantly. Spearman rank correlations showed that DMT1 vs. ferroportin, Dcytb vs. hephaestin and DMT1 vs. TFR1 mRNAs were positively correlated regardless of the underlying cause, similarly to protein levels of ferroportin vs. Dcytb and ferroportin vs. hephaestin. Serum ferritin was negatively correlated with DMT1 mRNA in investigated groups of patients, except for HHC group. A decrease of serum hepcidin was observed in IDA patients, but this was not statistically significant. Our data showed that although untreated HHC patients do not have increased mRNA levels of iron transport molecules when compared to normal subjects, the expression is relatively increased in relation to body iron stores. On the other hand, post-phlebotomized HHC patients had increased DMT1 and ferroportin mRNA levels possibly due to stimulated erythropoiesis after phlebotomy. Quelle: Journal of Cellular and Molecular Medicine | 5 Oct 2011 | 8:01 am CEST

Age-related changes in the contractile and passive arterial properties of murine mesenteric small arteries are altered by caveolin-1 knockout Abstract Caveolin-1, an integral protein of caveolae, is associated with multiple cardiovascular signalling pathways. Caveolin-1 knockout (KO) mice have a reduced lifespan. As changes in artery structure and function are associated with ageing we have investigated the role of caveolin-1 ablation on age-related changes of small artery contractility and passive mechanical properties. Mesenteric small arteries isolated from 3 and 12 month wild type (WT) and caveolin-1 KO mice were mounted on a pressure myograph and changes in passive and functional arterial properties were continuously monitored. In WT mice ageing was associated with a reduction in arterial contractility to noradrenaline which was reversed by inhibition of nitric oxide synthase (NOS) with L-NNA. Similarly, in 3 month old mice, caveolin-1 KO reduced contractility to noradrenaline by an L-NNA-sensitive mechanism. However, ageing in caveolin-1 KO mice was not associated with any further change in contractility. In WT mice ageing was associated with an increased passive arterial diameter and cross-sectional area (CSA), consistent with outward remodelling of the arterial wall, and a reduced arterial distensibility. Caveolin-1 ablation at 3 months of age resulted in similar changes in passive arterial properties to those observed with ageing in WT animals. However, ageing in caveolin-1 KO mice resulted in a reduced arterial CSA indicating different effects on passive structural characteristics from that observed in WT mice. Thus, caveolin-1 mice show abnormalities of small mesenteric artery function and passive mechanical characteristics indicative of premature vascular ageing. Moreover, caveolin-1 ablation modulates the age-related changes usually observed in mesenteric arteries of WT mice. Quelle: Journal of Cellular and Molecular Medicine | 5 Oct 2011 | 8:01 am CEST

TNF-α inhibits aquaporin 5 expression in human salivary gland acinar cells via suppression of histone H4 acetylation Abstract Sjögren's syndrome is a systemic autoimmune disease characterized by reductions in salivary and lacrimal secretions. The mechanisms underlying these reductions remain unclear. We have previously shown that TNF-α plays an important role in the destruction of acinar structures. Here we examined TNF-α's function in the expression of aquaporin (AQP) 5 in human salivary gland acinar cells. Immortalized human salivary gland acinar (NS-SV-AC) cells were treated with TNF-α, and then the expression levels of AQP5 mRNA and protein were analyzed. In addition, the mechanisms underlying the reduction of AQP5 expression by TNF-α treatment were investigated. TNF-α-treatment of NS-SV-AC cells significantly suppressed the expression levels of AQP5 mRNA and protein, and reduced the net fluid secretion rate. We examined the expression and activation levels of DNA methyltransferases (Dnmts) in NS-SV-AC cells treated with TNF-α. However, no significant changes were observed in the expression or activation levels of Dnmt1, Dnmt3a, or Dnmt3b. Although we also investigated the role of NF-κB activity in the TNF-α-induced suppression of AQP5 expression in NS-SV-AC cells, we detected similar TNF-α suppression of AQP5 expression in non-transfected cells and in a super-repressor form of IκBα cDNA-transfected cell clones. However, interestingly, chromatin immunoprecipitation analysis demonstrated a remarkable decrease in levels of acetylated histone H4 associated with the AQP5 gene promoter after treatment with TNF-α in NS-SV-AC cells. Therefore, our results may indicate that TNF-α inhibition of AQP5 expression in human salivary gland acinar cells is due to the epigenetic mechanism by suppression of acetylation of histone H4. Quelle: Journal of Cellular and Molecular Medicine | 5 Oct 2011 | 8:01 am CEST

Human versus porcine mesenchymal stromal cells: phenotype, differentiation potential, immunomodulation and cardiac improvement after transplantation Abstract Although Mesenchymal Stromal Cells (MSC) have been applied clinically to treat cardiac diseases, it is unclear how and to which extent transplanted MSC exert their beneficial effects. To address these questions, pre-clinical MSC administrations are needed for which pigs appear to be the species of choice. This requires the use of porcine cells to prevent immune rejection. However, it is currently unknown to what extent porcine MSC (pMSC) resemble human MSC (hMSC). Aim of this study was to compare MSC from porcine bone marrow (BM) with human cells for phenotype, multi-lineage differentiation potential, immune-modulatory capacity and the effect on cardiac function after transplantation in a mouse model of myocardial infarction. Flow cytometric analysis revealed that pMSC expressed surface antigens also found on hMSC, including CD90, MSCA-1 (TNAP/ W8B2 antigen), CD44, CD29, and SLA class I. Clonogenic outgrowth was significantly enriched following selection of CD271+ cells from BM of human and pig (129±29 and 1,961±485 fold, respectively). hMSC and pMSC differentiated comparably into the adipogenic, osteogenic or chondrogenic lineages, although pMSC formed fat much faster than hMSC. Immuno-modulation, an important feature of hMSC, was clearly demonstrated for pMSC when co-cultured with porcine peripheral blood cells stimulated with PMA and pIL-2. Finally, pMSC transplantation after myocardial infarction attenuated adverse remodeling to a similar extent as hMSC when compared to control saline injection. These findings demonstrate that pMSC have comparable characteristics and functionality with hMSC, making reliable extrapolation of pre-clinical pMSC studies into a clinical setting very well possible. Quelle: Journal of Cellular and Molecular Medicine | 5 Oct 2011 | 8:01 am CEST

Pathophysiology, staging and therapy of severe sepsis in baboon models Abstract We review our baboon models of E. coli sepsis that mimic, respectively, the shock/disseminated intravascular coagulation (DIC) and organ failure variants of severe sepsis, and analyze the pathophysiologic processes that are unique to each. The multistage, multifactorial characteristics of severe sepsis develop as a result of the initial insult, which -depending on its intensity- activates components of the intravascular compartment leading to overwhelming shock/DIC; or initiates a sequence of events involving both the intra- and extravascular (tissues) compartments that lead to organ failure. In the latter case, the disorder passes through two stages: an initial inflammatory/coagulopathic intravascular first stage triggered by E. coli, followed by an extravascular second stage, involving components unique to each organ and triggered by ischemia/reperfusion (oxidative stress and histone release). Though a myriad of overlapping cellular and molecular components are involved, it is the context in which these components are brought into play that determine whether shock/DIC or organ failure predominate. For example, inflammatory and thrombotic responses amplified by thrombin in the first case while similar responses are amplified by complement activation products in the second. Rather than blocking specific mediators, we found that attenuation of the thrombin and complement amplification pathways can effectively reverse the shock/DIC and organ failure exhibited by the LD100 and LD50E. coli models of severe sepsis, respectively. Translation of these concepts to successful intervention in the respective baboon models of E. coli sepsis and the application to their clinical counterparts is described. Quelle: Journal of Cellular and Molecular Medicine | 5 Oct 2011 | 8:01 am CEST

Ultrastructural differences between diabetic and idiopathic gastroparesis Abstract The ultrastructural changes in diabetic and idiopathic gastroparesis are not well studied and it is not known whether there are different defects in the two disorders. As part of the Gastroparesis Clinical Research Consortium, full thickness gastric body biopsies from 20 diabetic and 20 idiopathic gastroparetics were studied by light microscopy. Abnormalities were found in many (83%) but not all patients. Among the common defects were loss of interstitial cells of Cajal (ICC) and neural abnormalities. No distinguishing features were seen between diabetic and idiopathic gastroparesis. Our aim was to provide a detailed description of the ultrastructural abnormalities, compare findings between diabetic and idiopathic gastroparesis and determine if patients with apparently normal immunohistological features have ultrastructural abnormalities. Tissues from 40 gastroparetic patients and 24 age and sex matched controls were examined by transmission electron microscopy (TEM). ICC showing changes suggestive of injury, large and empty nerve endings, presence of lipofuscin and lamellar bodies in the smooth muscle cells were found in all patients. However, the ultrastructural changes in ICC and nerves differed between diabetic and idiopathic gastroparesis and were more severe in idiopathic gastroparesis. A thickened basal lamina around smooth muscle cells and nerves was characteristic of diabetic gastroparesis whereas idiopathic gastroparetics had fibrosis, especially around the nerves. In conclusion, in all the patients TEM showed abnormalities in ICC, nerves and smooth muscle consistent with the delay in gastric emptying. The significant differences found between diabetic and idiopathic gastroparesis offers insight into pathophysiology as well as into potential targeted therapies. Quelle: Journal of Cellular and Molecular Medicine | 14 Sep 2011 | 8:19 am CEST

Deep brain stimulation induces rapidly reversible transcript changes in Parkinson's leukocytes Abstract Sub-thalamic deep brain stimulation (DBS) reversibly modulates Parkinson's disease (PD) motor symptoms, providing an unusual opportunity to compare leukocyte transcripts in the same individuals before and after neurosurgery and one hour after stimulus cessation (ON- and OFF-stimulus). Here, we report DBS-induced reversibility and OFF-stimulus restoration in 12 of 16 molecular functions and 3 of 4 biological processes shown to be differentially expressed between PD patients and controls, post-DBS from pre-DBS and OFF from ON states. Intriguingly, 6 of 18 inflammation and immune-related functions exhibited reversibility, and the extent of stimulus-induced changes correlated with the neurological DBS efficacy, suggesting mechanistic implications. A minimal list of 29 transcripts that changed in all three comparisons between states discriminated pre-surgery and OFF states from post-surgery and controls. Six of these transcripts were found be able to distinguish between PD patients and both healthy controls and patients with other neurological diseases in a previously published whole blood 3' array data study of early PD patients.. Our findings support the future use of this approach for identifying targets for therapeutic intervention and assessing the efficacy of current and new treatments in this and other neurological diseases. Quelle: Journal of Cellular and Molecular Medicine | 12 Sep 2011 | 8:51 pm CEST

Elevation of IGF-2 receptor and the possible underlying implications in end-stage heart failure patients before and after heart transplantation Abstract Up-regulation of insulin-like growth factor 2 receptor (IGF-2R) involved in angiotensin II induced cell apoptosis in cardiomyoblasts, and correlated with cardiomyocyte apoptosis in hypertensive rat hearts. Here, we detected IGF-2R levels and explored the possible underlying implications in end-stage heart failure (HF) patients before and after heart transplantation. Western blot and imunohistochemistry were used to measure cardiac IGF-2R levels. Enzyme–linked immunosorbent assay (ELISA) was used to detect serum IGF-2R and CD8 levels. Labeling of DNA strand breaks and dihydroethidium detection were used to determine cellular apoptosis and reactive oxygen species, respectively. Cardiac IGF-2R levels increased in end-stage HF patients (n = 11) compared with non-failing control subjects. Leu27-IGF-2, an IGF-2 analog to activate specially the IGF-2R, could induce apoptosis and ROS production in neonatal rat ventricular myocytes. The serum IGF-2R levels were significantly higher in HF patients than those in non-failing control subjects. An unexpected observation is that the serum IGF-2R levels further increased after heart transplantation, peaked at the first month, and gradually reduced close to the levels before heart transplantation at the 6th months after heart transplantation. Serum CD8, a marker of acute rejection, had no change after heart transplantation, but IGF-2R and Granzyme B, as a ligand for the IGF-2R and a marker for CD8 T lymphocyte activation, coexisted in the transplanted hearts. Our preliminary studies suggest that elevation of IGF-2R may participate in pathological process of end-stage HF and involved in the acute cellular rejection after heart transplantation. Quelle: Journal of Cellular and Molecular Medicine | 7 Sep 2011 | 5:46 am CEST

PED/PEA-15 interacts with the 67 kDa laminin receptor and regulates cell adhesion, migration, proliferation and apoptosis Abstract Phosphoprotein enriched in diabetes/phosphoprotein enriched in astrocytes (PED/PEA-15) is an anti-apoptotic protein whose expression is increased in several human cancers. In addition to apoptosis, PED/PEA-15 is involved in the regulation of other major cellular functions, including cell adhesion, migration, proliferation and glucose metabolism. To further understand the functions of this protein, we performed a yeast two-hybrid screening using PED/PEA-15 as a bait and identified the 67 kDa high affinity laminin receptor (67LR) as an interacting partner. 67LR is a non-integrin cell-surface receptor for the extracellular matrix (ECM), derived from the dimerization of a 37 kDa cytosolic precursor (37LRP). The 67LR is highly expressed in human cancers and widely recognized as a molecular marker of metastatic aggressiveness. The molecular interaction of PED/PEA-15 with 67LR was confirmed by pull-down experiments with recombinant His-tagged 37LRP on lysates of PED/PEA-15 transfected HEK-293 cells. Further, overexpressed or endogenous PED/PEA-15 was co-immunoprecipitated with 67LR in PED/PEA-15 transfected HEK-293 cells and in U-373 glioblastoma cells, respectively. PED/PEA-15 overexpression significantly increased 67LR-mediated HEK-293 cell adhesion and migration to laminin that, in turn, determined PED/PEA-15 phosphorylation both in Ser-104 and Ser-116, thus enabling cell proliferation and resistance to apoptosis. PED/PEA-15 ability to induce cell responses to ECM-derived signals through interaction with 67LR may be of crucial importance for tumor cell survival in a poor microenvironment, thus favoring the metastatic spread and colonization. Quelle: Journal of Cellular and Molecular Medicine