ALBERT

Description:    INMAP was first identified as an interphase nucleus and mitotic apparatus-associated protein that plays essential roles in the formation of the spindle and cell-cycle progression. Here, we report that INMAP might be conserved from prokaryotes to humans, is a truncated version of the RNA polymerase III subunit B POLR3B, and is up-regulated in several human cancer cell lines including HeLa, Bel-7402, HepG2 and BGC-823. Deletion analysis revealed that the 209–290 amino-acid region is necessary for the punctate distribution of INMAP in the nucleus. Furthermore, over-expression of INMAP inhibited the transcriptional activities of p53 and AP-1 in a dose-dependent manner. These results suggest that INMAP may function through the p53 and AP-1 pathways, thus providing a possible link of its activity with tumourigenesis. Integrating our data and those in previous studies, it can be concluded that INMAP plays dual functional roles in the coordination of mitotic kinetics with gene expression as well as in cell-fate determination and proliferation. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1507-4 Authors Zhou Yunlei, Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education, College of Life Sciences, Beijing Normal University, Beijing, 100875 People’s Republic of China Chen Zhe, Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education, College of Life Sciences, Beijing Normal University, Beijing, 100875 People’s Republic of China Lei Yan, Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education, College of Life Sciences, Beijing Normal University, Beijing, 100875 People’s Republic of China Wang Pengcheng, Beijing Key Laboratory, College of Life Sciences, Beijing Normal University, Beijing, 100875 People’s Republic of China Zheng Yanbo, The Institute of Medical Biotechnology (IMB) of the Chinese Academy of Medical Sciences, Beijing, 100050 People’s Republic of China Sun Le, AbMax Biotechnology Co., Ltd, Haidian, Beijing, 100085 People’s Republic of China Liang Qianjin, Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education, College of Life Sciences, Beijing Normal University, Beijing, 100875 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The increased vascular calcification, cardiovascular morbidity, and mortality in chronic kidney disease (CKD) patients has been associated with disturbances in mineral-bone metabolism. In order to determine markers of the vascular calcification frequently observed in these patients, blood samples of elderly male and female hemodialysis CKD patients were used to measure serum levels of: osteoprotegerin (OPG), total soluble receptor activator of nuclear factor-κB ligand (sRANKL), and fetuin-A by enzyme immunoassay; tartrate-resistant acid phosphatase (TRACP-5b), and bone-specific alkaline phosphatase (BAP) by immunoenzymometric assay; osteocalcin (OC) by ELISA; iPTH by immunoradiometric assay; 25(OH)D 3 and 1,25(OH) 2 D 3 , by I 125 radioimmunoassay; and calcium and phosphorus by photometric assay. Serum OPG, BAP, iPTH, phosphorus, and OC levels were higher and serum 25(OH)D 3 , 1,25(OH) 2 D 3 , and fetuin-A levels lower in both male and female CKD patients than in their respective controls. Our results indicate that the bone formation and resorption parameters are altered in elderly male and female hemodialysis CKD patients. These changes may lead to vascular calcifications and cardiovascular complications, given that elevated OPG and OC levels and reduced fetuin-A levels are associated with cardiovascular events. Content Type Journal Article Pages 1-7 DOI 10.1007/s11010-012-1500-y Authors Alvaro Osorio, Vascular Surgery Unit, Sanitas Hospital, Madrid, Spain Esperanza Ortega, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Granada, Avenida de Madrid s/n, 18012 Granada, Spain Jesús M. Torres, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Granada, Avenida de Madrid s/n, 18012 Granada, Spain Pilar Sanchez, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Granada, Avenida de Madrid s/n, 18012 Granada, Spain Estrella Ruiz-Requena, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Granada, Avenida de Madrid s/n, 18012 Granada, Spain Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Hereditary haemorrhagic telangiectasia (HHT) is an autosomal dominant disease characterised by vascular dysplasia and increased bleeding that affect 1 in 5,000 people world-wide. Pathology is linked to mutations in genes encoding components of the heteromeric transforming growth factor-beta receptor (TGF-beta) and SMAD signalling pathway. Indeed HHT1 and HHT2 result from mutations in the genes encoding endoglin and activin-like kinase 1 (ALK1), TGF-beta receptor components. However, the fundamental cellular defects underlying HHT is poorly understood. Previously using confocal microscopy and N -glycosylation analysis, we found evidence that defective trafficking of endoglin from the endoplasmic reticulum (ER) to the plasma membrane is a mechanism underlying HHT1 in some patients. In this study, we used confocal microscopy to investigate whether a similar mechanism contributes to HHT2 pathology. To do this we expressed wild-type ALK1 and a number of HHT2 patient mutant variants as C-terminally tagged EGFP fusion proteins and tested their localisation in HeLa cells. We found that wild-type ALK1–EGFP was targeted predominantly to the plasma membrane, as evidenced by its colocalisation with the co-expressed HA-tagged endoglin. However, we found that in the majority of cases analysed the HHT2 patient mutant protein was retained within the ER as indicated by their colocalisation with the ER resident marker (calnexin) and lack of colocalisation with cell surface associated HA-endoglin. We conclude that defective trafficking and retention in the ER of mutant ALK1 protein is a possible mechanism of HHT2 in some patients. Content Type Journal Article Pages 1-11 DOI 10.1007/s11010-012-1496-3 Authors Alistair N. Hume, School of Biomedical Sciences, Queen’s Medical Centre, Nottingham, NG7 2UH UK Anne John, Departments of Pathology, College of Medicine and Health Sciences, United Arab Emirates University, P.O. Box 17666 Al-Ain, United Arab Emirates Nadia A. Akawi, Departments of Pathology, College of Medicine and Health Sciences, United Arab Emirates University, P.O. Box 17666 Al-Ain, United Arab Emirates Aydah M. Al-Awadhi, Departments of Pathology, College of Medicine and Health Sciences, United Arab Emirates University, P.O. Box 17666 Al-Ain, United Arab Emirates Sarah S. Al-Suwaidi, Departments of Pathology, College of Medicine and Health Sciences, United Arab Emirates University, P.O. Box 17666 Al-Ain, United Arab Emirates Lihadh Al-Gazali, Departments of Paediatrics, College of Medicine and Health Sciences, United Arab Emirates University, Al-Ain, United Arab Emirates Bassam R. Ali, Departments of Pathology, College of Medicine and Health Sciences, United Arab Emirates University, P.O. Box 17666 Al-Ain, United Arab Emirates Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Most studies using a hypercaloric diet to induce obesity have focused on the metabolism of fat and carbohydrates. Less concern has been given to the metabolism of amino acids, despite evidence of modifications in nitrogen metabolism during obesity. The aim of this study was to evaluate amino acid metabolism in livers from cafeteria diet-induced obese rats. Blood parameters were analysed, and histological sections of livers were stained with Sudan III. The enzymatic activities of some enzymes were determined in liver homogenates. Gluconeogenesis, ureagenesis, and oxygen consumption were evaluated in rat livers perfused with glutamine, alanine, or ammonium chloride. Compared to control rats, cafeteria-fed rats demonstrated higher levels of triacylglycerol and glucose in the blood and greater accumulation of fat in livers. Gluconeogenesis and urea production in livers perfused with glutamine and alanine at higher concentrations showed a substantial reduction in cafeteria-fed rats. However, no significant difference was observed among groups perfused with ammonium chloride. The activities of the enzymes alanine aminotransferase, glutaminase, and aspartate aminotransferase in the livers were reduced in cafeteria-fed rats. Taken together, these data are consistent with the hypothesis that livers from cafeteria diet-induced obese rats exhibit a limitation in their maximal capacity to metabolise glutamine and alanine to glucose, ammonia, and urea, not because of an impairment in gluconeogenesis and/or ureagenesis, but rather due to a depression in the activities of enzymes that catalyse the initial steps of amino acid metabolism. Content Type Journal Article Pages 1-13 DOI 10.1007/s11010-012-1499-0 Authors Cristiane Vizioli de Castro Ghizoni, Department of Biochemistry, Laboratory of Liver Metabolism, University of Maringá, Avenida Colombo 5790, 87020-900 Maringá, Paraná, Brazil Fabiana Rodrigues Silva Gasparin, Department of Biochemistry, Laboratory of Liver Metabolism, University of Maringá, Avenida Colombo 5790, 87020-900 Maringá, Paraná, Brazil Antonio Sueiti Maeda Júnior, Department of Biochemistry, Laboratory of Liver Metabolism, University of Maringá, Avenida Colombo 5790, 87020-900 Maringá, Paraná, Brazil Fernando Olinto Carreño, Department of Biochemistry, Laboratory of Liver Metabolism, University of Maringá, Avenida Colombo 5790, 87020-900 Maringá, Paraná, Brazil Rodrigo Polimeni Constantin, Department of Biochemistry, Laboratory of Liver Metabolism, University of Maringá, Avenida Colombo 5790, 87020-900 Maringá, Paraná, Brazil Adelar Bracht, Department of Biochemistry, Laboratory of Liver Metabolism, University of Maringá, Avenida Colombo 5790, 87020-900 Maringá, Paraná, Brazil Emy Luiza Ishii Iwamoto, Department of Biochemistry, Laboratory of Liver Metabolism, University of Maringá, Avenida Colombo 5790, 87020-900 Maringá, Paraná, Brazil Jorgete Constantin, Department of Biochemistry, Laboratory of Liver Metabolism, University of Maringá, Avenida Colombo 5790, 87020-900 Maringá, Paraná, Brazil Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The nasal epithelial barrier dysfunction is associated with the pathogenesis of nasal allergy; the causative factors are to be further elucidated. Ubiquitin E3 ligase TNFIAP3 (TNFIAP3, in short) plays a role in the maintenance of the homeostasis in the body. This study aims to elucidate the role of TNFIAP3 in the degradation of endocytic substances in nasal epithelial cells. The nasal epithelial cell line, RPMI 2650 cells (RPC), was cultured into monolayers in transwells. The endocytosis of staphylococcal enterotoxin B (SEB) by RPC monolayers was assessed by enzyme-linked immunoassay. The endocytosis of SEB-triggered endosome/lysosome fusion was observed by immunocytochemistry. The results showed that RPC monolayers expressed TNFIAP3 upon the endocytosis of SEB. Deficiency of TNFIAP3 resulted in abundant SEBs being transported to the basal chambers of transwells via the intracellular pathway. In the TNFIAP3-sufficient RPC, SEB-carrying endosomes fused with lysosomes were observed. The TNFIAP3-deficient RPC showed few SEB-carrying endosomes fused with lysosomes. In summary, TNFIAP3 plays an important role in tethering endosomes to lysosomes in RPC. Content Type Journal Article Pages 1-6 DOI 10.1007/s11010-012-1495-4 Authors Xingqiang Gao, Department of Otolaryngology & Head and Neck Surgery, First Hospital Affiliated Xiamen University, Xiamen, 361003 Fujian Province, China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Diabetes is associated with increased incidence of cardiovascular disease. Mechanisms that contribute to development of diabetic cardiopathy are not well understood. Phosphatidylinositol 3-kinase (PI3K) is a family of protein kinases that play an important role in regulation of cardiac function. It has been shown that inhibition of certain PI3K enzymes may produce cardiovascular protection. The aim of the present study was to determine whether chronic treatment with LY294002, an inhibitor of PI3K, can attenuate diabetes-induced cardiac dysfunction in isolated hearts obtained from normotensive and hypertensive rats. Recovery of cardiac function after 40 min of global ischemia and 30 min of reperfusion, measured as left ventricular developed pressure, left ventricular end-diastolic pressure, coronary flow and coronary vascular resistance, was worse in hearts obtained from diabetic and/or hypertensive animals compared to their respective controls. Treatment with LY294002 (1.2 mg/kg/day) for 4 weeks significantly prevented diabetes-induced cardiac dysfunction in both normotensive and hypertensive rats. Treatment with LY294002 did not significantly alter blood pressure or blood glucose levels. These results suggest that inhibition of PI3K signaling pathways can prevent ischemia/reperfusion-induced cardiac dysfunction in normotensive and hypertensive rats without correcting hyperglycemia or high blood pressure. Content Type Journal Article Pages 1-6 DOI 10.1007/s11010-012-1497-2 Authors Ibrahim F. Benter, Department of Pharmacology and Toxicology, Faculty of Medicine, Kuwait University, P.O. Box 24923, 13110 Safat, Kuwait Ibrahim Al-Rashdan, Department of Medicine, Faculty of Medicine, Kuwait University, P.O. Box 24923, 13110 Safat, Kuwait Jasbir S. Juggi, Faculty of Allied Health, Kuwait University, P.O. Box 24923, 13110 Safat, Kuwait Mariam H. M. Yousif, Department of Pharmacology and Toxicology, Faculty of Medicine, Kuwait University, P.O. Box 24923, 13110 Safat, Kuwait Saghir Akhtar, Department of Pharmacology and Toxicology, Faculty of Medicine, Kuwait University, P.O. Box 24923, 13110 Safat, Kuwait Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Recent studies have demonstrated that the Wnt/β-catenin signaling plays an important role in stem cell aging. However, the mechanisms of cell senescence induced by Wnt/β-catenin signaling are still poorly understood. Our preliminary study has indicated that activated Wnt/β-catenin signaling can induce MSC aging. In this study, we reported that the Wnt/β-catenin signaling was a potent activator of reactive oxygen species (ROS) generation in MSCs. After scavenging ROS with N -acetylcysteine, Wnt/β-catenin signaling-induced MSC aging was significantly attenuated and the DNA damage and the expression of p16 INK4A , p53, and p21 were reduced in MSCs. These results indicated that the Wnt/β-catenin signaling could induce MSC aging through promoting the intracellular production of ROS, and ROS may be the main mediators of MSC aging induced by excessive activation of Wnt/β-catenin signaling. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1498-1 Authors Da-yong Zhang, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Yu Pan, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Chong Zhang, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Bing-xi Yan, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Shan-shan Yu, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Dong-ling Wu, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Meng-meng Shi, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Kai Shi, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Xin-xiao Cai, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Shuang-shuang Zhou, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Jun-bo Wang, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Jian-ping Pan, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Li-huang Zhang, Department of Basic Medicine, School of Medicine and Life Sciences, Zhejiang University City College, 51 Huzhou Street, Hangzhou, 310015 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Hyperglycemia is the major cause of diabetic angiopathy. Sarpogrelate hydrochloride is an antiplatelet drug, and expected to be useful in the treatment of chronic arterial occlusive diseases. The aim of our study was to evaluate the possible effects of sarpogrelate hydrochloride on adhesion molecule expression and its underlying mechanism in the prevention and treatment of cardiovascular disorders. Intercellular adhesion molecule-1 (ICAM-1) expression and superoxide dismutase (SOD) activity were determined after endothelial cells were exposed to high glucose in the absence and presence of sarpogrelate hydrochloride. Coincubation of endothelial cells with high glucose for 24 h resulted in a significant increase of monocyte–endothelial cell adhesion and the expression of ICAM-1 ( P  〈 0.01). These effects were abolished by sarpogrelate hydrochloride and sarpogrelate hydrochloride significantly increased SOD activities (40 ± 8 vs. 47 ± 7, n  = 8, P  〈 0.01). The low dose sarpogrelate group (0.1 μM) had significantly higher monocyte–endothelial cell adhesion and the expression of ICAM-1 than medium dose sarpogrelate group (1.0 μM) and high dose sarpogrelate group (10.0 μM) ( P  〈 0.05 for comparison among three groups and P  〈 0.01 for difference between low and high dose sarpogrelate groups). These findings suggested that sarpogrelate hydrochloride was able to protect vascular endothelium from dysfunction induced by high glucose. Content Type Journal Article Pages 1-5 DOI 10.1007/s11010-012-1490-9 Authors Ying Su, Department of Endocrinology, The First Affiliated Hospital of Harbin Medical University, No. 23 Youzheng Street, NanGang, Harbin, 150001 China Nan Mao, Department of Endocrinology, The First Affiliated Hospital of Harbin Medical University, No. 23 Youzheng Street, NanGang, Harbin, 150001 China Min Li, Department of Endocrinology, The First Affiliated Hospital of Harbin Medical University, No. 23 Youzheng Street, NanGang, Harbin, 150001 China Xia Dong, Department of Endocrinology, The First Affiliated Hospital of Harbin Medical University, No. 23 Youzheng Street, NanGang, Harbin, 150001 China Fan-Zhen Lin, Department of Endocrinology, The First Affiliated Hospital of Harbin Medical University, No. 23 Youzheng Street, NanGang, Harbin, 150001 China Ying Xu, Department of Endocrinology, The First Affiliated Hospital of Harbin Medical University, No. 23 Youzheng Street, NanGang, Harbin, 150001 China Yan-Bo Li, Department of Endocrinology, The First Affiliated Hospital of Harbin Medical University, No. 23 Youzheng Street, NanGang, Harbin, 150001 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    In addition of being an important inflammatory biomarker and a risk factor for cardiovascular disease, much evidence indicates that the C-reactive protein (CRP) contributes to the atherosclerosis development process. This plasmatic protein synthesized by hepatocytes in response to inflammation and tissue injury induces pro-inflammatory molecules' expression by endothelial cells (ECs). Previous studies showed that the 17β-estradiol (E2) has beneficial effects on vascular cells by reducing in vitro pro-inflammatory molecules expressions in EC. Therefore, we hypothesize that E2 blocks or reduces CRP-mediated inflammatory responses by modulating endogenous production of CRP in EC and/or activation mechanisms. Using human aortic ECs (HAECs), we first evaluated CRP production by vascular EC and second demonstrated its self-induction. Indeed, recombinant human CRP stimulation induces a fivefold increase of CRP expression. A 1-h pre-treatment of E2 at a physiologic dose (10 −9  M) leads to an important decrease of CRP production suggesting a partial blockage of its amplification loop mechanism. Furthermore, in HAEC, E2 reduces the secretion of the most potent agonist of CRP induction, the IL-6, by 21 %. E2 pre-treatment also decreased the expression of pro-inflammatory molecules IL-8, VCAM-1, and ICAM-1 induced by CRP and involved in leukocytes recruitment. In addition, we demonstrated that E2 could restore vascular endothelial growth factor-mediated EC migration response impaired by CRP suggesting another pro-angiogenic property of this hormone. These findings suggest that E2 can interfere with CRP pro-inflammatory effects via activation signals using its rapid, non-genomic pathway that may provide a new mechanism to improve vascular repair. Content Type Journal Article Pages 1-11 DOI 10.1007/s11010-012-1482-9 Authors Émilie Cossette, Research Center, Montreal Heart Institute, 5000 Bélanger Street, Montreal, QC H1T 1C8, Canada Isabelle Cloutier, Research Center, Montreal Heart Institute, 5000 Bélanger Street, Montreal, QC H1T 1C8, Canada Kim Tardif, Research Center, Montreal Heart Institute, 5000 Bélanger Street, Montreal, QC H1T 1C8, Canada Geneviève DonPierre, Research Center, Montreal Heart Institute, 5000 Bélanger Street, Montreal, QC H1T 1C8, Canada Jean-François Tanguay, Research Center, Montreal Heart Institute, 5000 Bélanger Street, Montreal, QC H1T 1C8, Canada Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Sarcoplasmic and t-tubule membrane proteins regulating sarcoplasmic Ca 2+ concentration exhibit fibre-type-dependent isoform expression, and play central roles in muscle contraction and relaxation. The purpose of this study was to evaluate the effects of in vitro electrical stimulation on the mRNA expression of components involved in Ca 2+ regulation in oxidative and glycolytic skeletal muscle. The mRNA level of Ca 2+ -ATPase (SERCA1, 2), calsequestrin (CASQ1, 2), ryanodine receptor (RyR1), and dihydropyridine receptor (Cacna1) was assessed in rat extensor digitorum longus (EDL) and soleus (SOL) muscles at 4 h of recovery following in vitro stimulations (either short intensive (SHO) 60 Hz, 5 min, or prolonged moderate (PRO) 20 Hz, 40 min). Stimulation induced acute regulation of the mRNA level of Ca 2+ -regulating proteins in a manner that does not follow typical fibre-type-specific transitions. In general, stimulation decreased mRNA content of all proteins studied. Most prominent down-regulation was observed for Cacna1 (26 and 32 % after SHO and PRO, respectively, in SOL; 19 % after SHO in EDL). SERCA1, SERCA2, CASQ1, CASQ2, and RyR1 mRNA content also decreased significantly in both muscles relative to resting control. Of notice is that hexokinase II mRNA content was increased in EDL and unchanged in SOL underlining the specificity of the down-regulation of mRNA of Ca 2+ regulatory proteins. The results demonstrate contraction-induced down-regulation of mRNAs for the main components of Ca 2+ -regulating system in skeletal muscle. The down-regulation of both isoforms of SERCA and CASQ after a single electrical stimulation session suggests that adaptations to repeated stimulation involve further regulatory mechanisms in addition to acute mRNA responses. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1486-5 Authors Satu Mänttäri, Department of Biology, University of Oulu, P.O. Box 3000, 90014 Oulu, Finland Niels Ørtenblad, Institute of Sports Science and Clinical Biomechanics, University of Southern Denmark, 5230 Odense, Denmark Klavs Madsen, Department of Public Health—Sport Science, University of Aarhus, 8000 Aarhus, Denmark Henriette Pilegaard, Department of Biology, Centre of Inflammation and Metabolism and Copenhagen Muscle Research Centre, University of Copenhagen, 2199 Copenhagen, Denmark Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    SASH1, a member of the SLY-family of signal adapter proteins, is a candidate tumor suppressor in breast and colon cancer. The SASH1 protein possesses both the SH3 and SAM domains, indicating that it may play an important role in intracellular signal transduction. Reduced expression of SASH1 is closely related to tumor growth, invasion, metastasis, and poor prognosis. However, the biological role of SASH1 remains unknown in osteosarcoma. To unravel the function of SASH1, we explored the expression of SASH1 in osteosarcoma tissues and its correlation to the clinical pathology of osteosarcoma and analyzed the relationship between SASH1 expression and cell cycle, apoptosis and invasion of osteosarcoma MG-63 cells, using the flow cytometry analysis and transwell invasion chamber experiments. Furthermore, the effect of SASH1 on the expression of cyclin D1, caspase-3, matrix metalloproteinase (MMP)-9 were observed by western blot. Our results showed that the expression rate of SASH1 mRNA in osteosarcoma tissues was significantly lower than that in normal bone tissue ( p  = 0.000), that the expression rate of SASH1 mRNA in the carcinoma tissues from patients with lung metastasis was significantly lower than that from patients without lung metastasis ( p  = 0.041), and that the expression rate of SASH1 mRNA also decreased with increasing Enneking stage ( p  = 0.032). However, the mRNA expression of SASH1 in osteosarcoma was independent of the patient’s gender, age, and tumor size ( p  = 0.983, 0.343, 0.517, respectively). The SASH1 protein displayed a down-regulation in osteosarcoma tissues compared to normal bone tissue ( p  = 0.000), displayed a down-regulation in osteosarcoma tissues from patients with lung metastasis compared to from patients without lung metastasis ( p  = 0.000), and displayed a gradual decrease with increasing Enneking stage ( p  = 0.000). In addition, the MG-63 cells from pcDNA3.1-SASH1 group exhibited significantly reduced cell viability, proliferation, and invasive ability compared to the empty vector group and blank control group ( p  = 0.023, 0.001, respectively), and there was no difference between the empty vector group and blank control group. The pcDNA3.1-SASH1 group displayed significantly more apoptotic cells than the empty vector group and blank control group ( p  = 0.004). The expression of cyclin D1, MMP-9 displayed a down-regulation in MG-63 cells from pcDNA3.1-SASH1 group compared to the empty vector group and blank control group ( p  = 0.000, 0.001, respectively) and the expression levels of caspase-3 displayed an up-regulation in MG-63 cells from pcDNA3.1-SASH1 group compared to the empty vector group and blank control group ( p  = 0.000). Taken together, these data indicated that the overexpression of SASH1 might be associated with the inhibition of growth, proliferation, and invasion of MG-63 cells and the promotion of apoptosis of MG-63 cells. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1491-8 Authors Qingbing Meng, Orthopedics Department, Yancheng City No. 1 People’s Hospital, 16 Yue-He Road, Yancheng, 224005 Jiangsu, People’s Republic of China Minqian Zheng, Orthopedics Department, Yancheng City No. 1 People’s Hospital, 16 Yue-He Road, Yancheng, 224005 Jiangsu, People’s Republic of China Hongbing Liu, Orthopedics Department, Yancheng City No. 1 People’s Hospital, 16 Yue-He Road, Yancheng, 224005 Jiangsu, People’s Republic of China Changzhi Song, Orthopedics Department, Yancheng City No. 1 People’s Hospital, 16 Yue-He Road, Yancheng, 224005 Jiangsu, People’s Republic of China Wensheng Zhang, Orthopedics Department, Yancheng City No. 1 People’s Hospital, 16 Yue-He Road, Yancheng, 224005 Jiangsu, People’s Republic of China Juan Yan, Orthopedics Department, Yancheng City No. 1 People’s Hospital, 16 Yue-He Road, Yancheng, 224005 Jiangsu, People’s Republic of China Ling Qin, Orthopedics Department, Yancheng City No. 1 People’s Hospital, 16 Yue-He Road, Yancheng, 224005 Jiangsu, People’s Republic of China Xiaolan Liu, Orthopedics Department, Yancheng City No. 1 People’s Hospital, 16 Yue-He Road, Yancheng, 224005 Jiangsu, People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The present study was designed to evaluate the in vitro and in vivo ameliorative antioxidant potential of secoisolariciresinol diglucoside (SDG). In vitro antioxidant activity of synthetic SDG was carried out using DPPH, reducing power potency, and DNA protection assays. Wistar albino rats weighing 180–220 g were used for in vivo studies and liver damage was induced in the experimental animals by a single intraperitoneal (I.P.) injection of CCl 4 (2 g/kg b.w.). Intoxicated animals were treated orally with synthetic SDG at (12.5 and 25 mg/kg b.w.) and Silymarin (25 mg/kg) for 14 consecutive days. The levels of catalase (CAT), superoxide dismutase (SOD), peroxidase (POX), and lipid peroxidase (LPO) were measured in liver and kidney homogenates. The synthetic SDG exerts high in vitro antioxidant potency as it could scavenge DPPH at a IC 50 value of 78.9 μg/ml and has dose-dependent reducing power potency and protected DNA at 0.5 mg/ml concentration. Oral administration of synthetic SDG at 12.5 and 25 mg/kg b.w. showed significant protection compared to Silymarin (25 mg/kg) and the activities of CAT, SOD, and POX were markedly increased ( P  〈 0.05), whereas LPO significantly decreased ( P  〈 0.001) in a dose-dependent manner in liver and kidney in both pre- and post-treatment groups when compared to toxin-treated group. The results of in vitro and in vivo investigations revealed that synthetic SDG at 25 mg/kg b.w. is associated with beneficial changes in hepatic enzyme activities and thereby plays a key role in the prevention of oxidative damage in immunologic system. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1487-4 Authors Sadiq S. Moree, Department of Biochemistry, Yuvaraja’s College, University of Mysore, Mysore, 570005 Karnataka, India J. Rajesha, Department of Biochemistry, Yuvaraja’s College, University of Mysore, Mysore, 570005 Karnataka, India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Marked sex-dependent differences in mitochondrial function and redox status have been found in brown adipose tissue (BAT) of control rats. Insulin also plays a role in the development and maintenance of this tissue. The aim was to investigate sexual dimorphism in the effects of diet-induced obesity on BAT mitochondrial function, as well as on insulin signaling pathway. 10-week-old Wistar rats of both sexes were fed a control diet or a palatable high-fat diet for 26 weeks. Serum markers of insulin sensitivity were analyzed. Mitochondrial DNA (mtDNA) content, mitochondrial oxidative activities, PGC-1α mRNA levels, as well as the protein levels of insulin receptor subunit β (IRβ), glucose transporter GLUT4, β 3 -adrenergic receptor (β 3 -AR), phosphatidylinositol 3-kinase, mitochondrial transcription factor A (TFAM), cytochrome c oxidase subunit IV (COX IV), and uncoupling protein 1 (UCP1) were measured in BAT. Obese females showed impaired systemic insulin sensitivity accompanied by diminished IRβ, GLUT4, and β 3 -AR protein levels in BAT. In addition, TFAM and COX IV protein and PGC-1α mRNA levels decreased in obese females, whereas mtDNA levels increased. In obese males, oxidative and thermogenic capacities rose and no significant changes were observed in the insulin signaling pathway elements. The reduction of the insulin signaling pathway in BAT of obese females may be responsible, at least partially, for the impaired biogenesis process, which could favor the increase of body weight found in this sex. In contrast, the enhanced mitochondrial functionality in the BAT of males would avoid increased oxidative damage and the impairment of insulin signaling. Content Type Journal Article Pages 1-11 DOI 10.1007/s11010-012-1481-x Authors A. Nadal-Casellas, Grup de Metabolisme Energètic i Nutrició, Departament de Biologia Fonamental i Ciències de la Salut, Institut Universitari d’Investigació en Ciències de la Salut (IUNICS), Universitat de les Illes Balears, Cra. Valldemossa km 7.5, 07122 Palma, Spain M. Bauzá-Thorbrügge, Grup de Metabolisme Energètic i Nutrició, Departament de Biologia Fonamental i Ciències de la Salut, Institut Universitari d’Investigació en Ciències de la Salut (IUNICS), Universitat de les Illes Balears, Cra. Valldemossa km 7.5, 07122 Palma, Spain A. M. Proenza, Grup de Metabolisme Energètic i Nutrició, Departament de Biologia Fonamental i Ciències de la Salut, Institut Universitari d’Investigació en Ciències de la Salut (IUNICS), Universitat de les Illes Balears, Cra. Valldemossa km 7.5, 07122 Palma, Spain M. Gianotti, Grup de Metabolisme Energètic i Nutrició, Departament de Biologia Fonamental i Ciències de la Salut, Institut Universitari d’Investigació en Ciències de la Salut (IUNICS), Universitat de les Illes Balears, Cra. Valldemossa km 7.5, 07122 Palma, Spain I. Lladó, Grup de Metabolisme Energètic i Nutrició, Departament de Biologia Fonamental i Ciències de la Salut, Institut Universitari d’Investigació en Ciències de la Salut (IUNICS), Universitat de les Illes Balears, Cra. Valldemossa km 7.5, 07122 Palma, Spain Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    To investigate the combined effect of aliskiren, a renin inhibitor, and AVE 0991, a Mas-receptor agonist, in experimental hypertension (HT) in rats. HT was produced by administration of deoxycorticosterone acetate (DOCA) and mean arterial blood pressure (MABP) was assessed by tail-cuff method. Treatments were started from 4th week onwards and were continued for 9 days. A significant increase in MABP was noted after 1 week in DOCA control rats, as compared with the base line value. A stable HT developed after 4 weeks of DOCA administration. Treatments with aliskiren and AVE 0991 alone, dose-dependently decreased MABP in DOCA-treated rats. Further, combination of low doses of aliskiren and AVE 0991 significantly reduced MABP, as compared with DOCA control rats and with either drug alone in low doses. It may be concluded that treatment with aliskiren produced down-regulation of both harmful Ang II–AT1-receptor and survival Ang(1–7)/Mas-receptor axis of RAAS. Treatment with combination of low doses of aliskiren and AVE 0991, for the first time, has been shown to produce synergistic blood pressure lowering effect. Therefore, combination of renin inhibitor with Mas-receptor agonist may prove beneficial for the treatment of hypertensive patients. Content Type Journal Article Pages 1-6 DOI 10.1007/s11010-012-1489-2 Authors Yogendra Singh, Department of Pharmacology, ISF College of Pharmacy, Moga, 142 001 Punjab, India Kulwinder Singh, University Institute of Pharmacy, Baba Farid University of Health Sciences, Faridkot, 151 203 Punjab, India P. L. Sharma, Department of Pharmacology, ISF College of Pharmacy, Moga, 142 001 Punjab, India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Sulforaphane (SFN), a component of dietary cruciferous vegetables has been characterized for its anti-proliferative properties. We have recently demonstrated that pancreatic CSCs display activation of sonic hedgehog pathway which are fundamental drivers of stem cell renewal, and SFN inhibits the self-renewal of pancreatic CSCs in vitro. Consistent with these observations, we sought to determine the chemopreventive potential of SFN in an in vivo setting. We show here for the first time that sulforaphane treatment resulted in a significant reduction in the tumor growth of orthotopically implanted primary pancreatic CSCs isolated from human pancreatic tumors into the pancreas of NOD/SCID/IL2Rgamma mice, which is mediated through the modulation of Sonic hedgehog–GLI signaling. Hedgehog pathway blockade by SFN at a dose of 20 mg/kg resulted in a 45 % reduction in growth of pancreatic cancer tumors and reduced expression of Shh pathway components, Smo, Gli 1, and Gli 2 in mouse tissues. Further, SFN inhibited the expression of pluripotency maintaining transcription factors Nanog and Oct-4 and angiogenic markers VEGF and PDGFRα which are downstream targets of Gli transcription. Furthermore, SFN treatment resulted in a significant reduction in EMT markers Zeb-1, which correlated with increase in E-Cadherin expression suggesting the blockade of signaling involved in early metastasis. Interestingly, SFN downregulated the expression of Bcl-2 and XIAP to induce apoptosis. These data demonstrate that, at a tolerable dose, inhibition of Shh pathway by SFN results in marked reduction in EMT, metastatic, angiogenic markers with significant inhibition in tumor growth in mice. Since aberrant Shh signaling occurs in pancreatic tumorigenesis, therapeutics that target Shh pathway may improve the outcomes of patients with pancreatic cancer by targeting CSCs, thus suggesting the use of sulforaphane to further improve preventive and therapeutic approaches in patients with this devastating disease. Content Type Journal Article Pages 1-11 DOI 10.1007/s11010-012-1493-6 Authors Shih-Hui Li, Department of Pathology and Laboratory Medicine, The University of Kansas Cancer Center, The University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, KS 66160, USA Junsheng Fu, Department of Pathology and Laboratory Medicine, The University of Kansas Cancer Center, The University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, KS 66160, USA Dara Nall Watkins, Department of Pathology and Laboratory Medicine, The University of Kansas Cancer Center, The University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, KS 66160, USA Rakesh K. Srivastava, Department of Pharmacology, Toxicology and Therapeutics, and Medicine, The University of Kansas Cancer Center, The University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, KS 66160, USA Sharmila Shankar, Department of Pathology and Laboratory Medicine, The University of Kansas Cancer Center, The University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, KS 66160, USA Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Dynamic remodeling of the actin cytoskeleton is crucial for biological processes such as cell migration and cell spreading. S100A10 is a member of the S100 protein family and is involved in intracellular trafficking and cell migration. In this study, we examined the role of S100A10 in actin cytoskeletal organization and cell spreading. Depletion of S100A10 induced disruption of stress fiber formation and delay in cell spreading. Rac1 activation during spreading was suppressed by S100A10 knockdown, and exogenous expression of active Rac1 restored the ability of cells to spread in the absence of S100A10. Our results demonstrate the crucial role of S100A10 in actin dynamics promoting cell spreading via Rac1 activation. Content Type Journal Article Pages 1-7 DOI 10.1007/s11010-012-1509-2 Authors Shurovi Sayeed, Division of Cancer Biology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550 Japan Eri Asano, Division of Cancer Biology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550 Japan Satoko Ito, Division of Cancer Biology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550 Japan Kinji Ohno, Division of Neurogenetics, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550 Japan Michinari Hamaguchi, Division of Cancer Biology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550 Japan Takeshi Senga, Division of Cancer Biology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550 Japan Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Chronic hepatitis B virus infection is the dominant global cause of hepatocellular carcinoma (HCC), especially hepatitis B virus-X (HBx) plays a major role in this process. HBx protein promotes cell cycle progression, inactivates negative growth regulators, and binds to and inhibits the expression of p53 tumor suppressor gene and other tumor suppressor genes and senescence-related factors. However, the relationship between HBx and autophagy during the HCC development is poorly known. Previous studies found that autophagy functions as a survival mechanism in liver cancer cells. We suggest that autophagy plays a possible role in the pathogenesis of HBx-induced HCC. The present study showed that HBx transfection brought about an increase in the formation of autophagosomes and autolysosomes. Microtubule-associated protein light chain 3, Beclin 1, and lysosome-associated membrane protein 2a were up-regulated after HBx transfection. HBx-induced increase in the autophagic level was increased by mTOR inhibitor rapamycin and was blocked by treatment with the PI3K–Akt inhibitor LY294002. The same results can also be found in HepG2.2.15 cells. These results suggest that HBx activates the autophagic lysosome pathway in HepG-2 cells through the PI3K–Akt–mTOR pathway. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1457-x Authors Peng Wang, Department of Hepatobiliary Surgery, First Affiliated Hospital of Soochow University, Suzhou, 215004 People’s Republic of China Qing-song Guo, Department of Hepatobiliary Surgery, Affiliated Hospital of Nantong University, Nantong, 226001 People’s Republic of China Zhi-wei Wang, Department of Hepatobiliary Surgery, Affiliated Hospital of Nantong University, Nantong, 226001 People’s Republic of China Hai-xin Qian, Department of Hepatobiliary Surgery, First Affiliated Hospital of Soochow University, Suzhou, 215004 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Emerging evidences demonstrate that excess aldosterone and insulin interact at target tissues. It has been shown that increased levels of aldosterone contribute to the development of insulin resistance and thus act as a risk factor for the development of type-2 diabetes mellitus. However, the molecular mechanisms involved in this scenario are yet to be identified. This study was designed to assess the dose-dependent effects of aldosterone on insulin signal transduction and glucose oxidation in the skeletal muscle (gastrocnemius) of adult male rat. Healthy adult male albino rats of Wistar strain ( Rattus norvegicus ) weighing 180–200 g were used in this study. Rats were divided into four groups. Group I: control (treated with 1 % ethanol only), group II: aldosterone treated (10 μg /kg body weight, twice daily for 15 days), group III: aldosterone treated (20 μg /kg body weight, twice daily for 15 days), and group IV: aldosterone treated (40 μg/kg body weight, twice daily for 15 days). Excess aldosterone caused glucose intolerance in a dose-dependent manner. Serum insulin and aldosterone were significantly increased, whereas serum testosterone was decreased. Aldosterone treatment impaired the rate of glucose uptake, oxidation, and insulin signal transduction in the gastrocnemius muscle through defective expression of IR, IRS-1, Akt, AS160, and GLUT4 genes. Phosphorylation of IRS-1, β-arrestin-2, and Akt was also reduced in a dose-dependent manner. Excess aldosterone results in glucose intolerance as a result of impaired insulin signal transduction leading to decreased glucose uptake and oxidation in skeletal muscle. In addition to this, it is inferred that excess aldosterone may act as one of the causative factors for the onset of insulin resistance and thus increased incidence of type-2 diabetes. Content Type Journal Article Pages 1-14 DOI 10.1007/s11010-012-1452-2 Authors Jayaraman Selvaraj, Department of Endocrinology, Dr.ALM Post Graduate Institute of Basic Medical Sciences, University of Madras, Sekkizhar Campus, Taramani, Chennai, 600113 India Sampath Sathish, Department of Endocrinology, Dr.ALM Post Graduate Institute of Basic Medical Sciences, University of Madras, Sekkizhar Campus, Taramani, Chennai, 600113 India Chinnaiyan Mayilvanan, Department of Endocrinology, Dr.ALM Post Graduate Institute of Basic Medical Sciences, University of Madras, Sekkizhar Campus, Taramani, Chennai, 600113 India Karundevi Balasubramanian, Department of Endocrinology, Dr.ALM Post Graduate Institute of Basic Medical Sciences, University of Madras, Sekkizhar Campus, Taramani, Chennai, 600113 India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Activation of protein kinase C (PKC) is a critical intracellular signaling triggered by ischemic preconditioning (IPC), but the precise mechanisms underlying the actions of PKC in IPC-mediated cardioprotection remain unclear. Here, we investigated the role of PKC activation on the antioxidant activity by IPC in rabbit hearts. Isolated rabbit hearts were subjected to 60 min of global ischemia by cold cardioplegic arrest (4 °C) and 60 min of reperfusion (37 °C). IPC was induced by three cycles of 2-min ischemia following 3 min of reperfusion (37 °C) before cardioplegic arrest. IPC resulted in a better recovery of mechanical function, increased tissue reduced glutathione-to-oxidized glutathione ratio (GSH/GSSG), superoxide dismutase and catalase content, and decreased tissue malondialdehyde (MDA) content compared to control hearts subjected to 60 min of cardioplegic ischemia and 60 min of reperfusion. IPC also significantly induced activation of nuclear factor erythroid 2-related factor 2 (Nrf2) and the inductions of antioxidant genes heme oxygenase-1 (HO-1) and manganese superoxide dismutase (MnSOD). Injection of phorbol 12-myristate 13 acetate, an activator of PKC, before cardioplegic ischemia induced translocation of PKC-δ and -ε isoforms to membrane fraction, nuclear accumulation of Nrf2, and conferred cardioprotection similar to IPC. Polymyxin B, an inhibitor of PKC, blocked the membrane translocation of PKC-δ and -ε during IPC, inhibited Nrf2 nuclear accumulation, and significantly diminished the IPC-induced cardioprotection when administrated before IPC. These results indicate that the activation of PKC induces the translocation of Nrf2 and the enhancement of endogenous antioxidant defenses in the IPC hearts and suggest that PKC may target Nrf2 to confer cardioprotection. Content Type Journal Article Pages 1-11 DOI 10.1007/s11010-012-1458-9 Authors Xin Zhang, Department of Cardiac Surgery, Zhengzhou University College of Medicine, Zhengzhou, 450052 China Zhibin Xiao, Department of Cardiac Surgery, Beijing General Hospital of PLA, Beijing, 100700 China Jianmin Yao, Department of Cardiac Surgery, Beijing General Hospital of PLA, Beijing, 100700 China Genshang Zhao, Department of Cardiac Surgery, Zhengzhou University College of Medicine, Zhengzhou, 450052 China Xianen Fa, Department of Cardiac Surgery, Zhengzhou University College of Medicine, Zhengzhou, 450052 China Jianli Niu, Department of Cardiac Surgery, Beijing General Hospital of PLA, Beijing, 100700 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Serotonin (5-HT) is a hormone that has been implicated in the regulation of many physiological and pathological events. One of the most intriguing properties of this hormone is its ability to up-regulate mitosis. Moreover, 5-HT stimulates glucose uptake and up-regulates PFK activity through the 5-HT 2A receptor, resulting in the phosphorylation of a tyrosine residue of PFK and the intracellular redistribution of PFK within skeletal muscle. The present study investigated some of the signaling intermediates involved in the effects of 5-HT on 6-phosphofructo-1-kinase (PFK) regulation from skeletal muscle using kinetic assessments, immunoprecipitation, and western blotting assays. Our results demonstrate that 5-HT stimulates PFK from skeletal muscle via phospholipase C (PLC). The activation of PLC in skeletal muscle leads to the recruitment of protein kinase C (PKC) and calmodulin and the stimulation of calmodulin kinase II, which associates with PFK upon 5-HT action. Alternatively, 5-HT loses its ability to up-regulate PFK activity when Janus kinase is inhibited, suggesting that 5-HT is able to control glycolytic flux in the skeletal muscle of mice by recruiting different pathways and controlling PFK activity. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1462-0 Authors Wagner Santos Coelho, Laboratório de Enzimologia e Controle do Metabolismo (LabECoM), Departamento de Fármacos, Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Ilha do Fundão, Rio de Janeiro, RJ 21941-590, Brazil Mauro Sola-Penna, Laboratório de Enzimologia e Controle do Metabolismo (LabECoM), Departamento de Fármacos, Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Ilha do Fundão, Rio de Janeiro, RJ 21941-590, Brazil Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Upon release from keratinocytes, 14-3-3 sigma (also known as stratifin) acts on the dermal fibroblast and modulates its production of extracellular matrix proteins. Subsequent to the recent identification as a receptor responsible for stratifin-mediated matrix turnover in dermal fibroblasts, aminopeptidase N has been implicated in the regulation of epidermal–dermal communication and expression of key matrix proteases and adhesion molecules. In light of the growing importance of aminopeptidase N in modulation of the fibroblast phenotype, the present study evaluates the potential of targeting the ectoenzyme in cutaneous repair, and demonstrates that neutralization of aminopeptidase N led to acceleration of wound closure. This was attributed to at least in part an increase of collagen deposition and fibroblast contractility in the granulation tissue. These findings confirmed the important role of aminopeptidase N in post-injury tissue remodeling and wound contraction. Content Type Journal Article Pages 1-6 DOI 10.1007/s11010-012-1449-x Authors Amy Lai, Burn and Wound Healing Research Laboratory, ICORD, the Blusson Spinal Cord Centre, Department of Surgery, University of British Columbia, 818 West 10th Avenue, Vancouver, BC V5Z 1M9, Canada Azadeh Hosseini-Tabatabaei, Burn and Wound Healing Research Laboratory, ICORD, the Blusson Spinal Cord Centre, Department of Surgery, University of British Columbia, 818 West 10th Avenue, Vancouver, BC V5Z 1M9, Canada Ryan Hartwell, Burn and Wound Healing Research Laboratory, ICORD, the Blusson Spinal Cord Centre, Department of Surgery, University of British Columbia, 818 West 10th Avenue, Vancouver, BC V5Z 1M9, Canada Elham Rahmani-Neishaboor, Burn and Wound Healing Research Laboratory, ICORD, the Blusson Spinal Cord Centre, Department of Surgery, University of British Columbia, 818 West 10th Avenue, Vancouver, BC V5Z 1M9, Canada Ruhangiz Taghi Kilani, Burn and Wound Healing Research Laboratory, ICORD, the Blusson Spinal Cord Centre, Department of Surgery, University of British Columbia, 818 West 10th Avenue, Vancouver, BC V5Z 1M9, Canada Aziz Ghahary, Burn and Wound Healing Research Laboratory, ICORD, the Blusson Spinal Cord Centre, Department of Surgery, University of British Columbia, 818 West 10th Avenue, Vancouver, BC V5Z 1M9, Canada Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    NF-κB consists of p50, p65 (RelA), p52, c-Rel, and RelB, and among them p65 is a representative protein to investigate the regulation and function of this signaling. NF-κB integrates inflammation and carcinogenesis and regulates the expression of a variety of genes in response to immunity, inflammation, and apoptosis. IκBα acts as an inhibitor of NF-κB through forming an inactive NF-κB/IκBα complex. Pokemon is a ubiquitous transcription factor involved in different signaling pathways, playing a pivotal role in cell proliferation, anti-apoptosis, embryonic development, and maintenance. In this study, we found that p65 and IκBα are both novel regulatory targets of Pokemon. Ectopic expression of Pokemon in immortalized liver cells HL7702 enhanced p65 and IκBα expression, whereas silencing of Pokemon in hepatocellular carcinoma cells QGY7703 reduced cellular p65 levels. ChIP assay and targeted mutagenesis revealed that Pokemon directly binds to the element of −434 to −430 bp in p65 promoter and of −453 to −448 bp in IκBα promoter and stimulates luciferase reporter gene expression. Co-transfection of Pokemon with p65 or IκBα promoter-reporter notably enhanced their promoter activity. These data suggest that Pokemon activates the expression of both p65 and IκBα by sequence-specific binding to their promoters and plays a dual role in regulating NF-κB signaling. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1445-1 Authors Nan-Nan Zhang, Department of Chemistry, Tsinghua University, Beijing, 100084 People’s Republic of China Qin-Sheng Sun, The Ministry-Province Jointly Constructed Base for State Key Lab-Shenzhen Key Laboratory of Chemical Biology, the Graduate School at Shenzhen, Tsinghua University, Shenzhen, 518055 People’s Republic of China Zhe Chen, Department of Chemistry, Tsinghua University, Beijing, 100084 People’s Republic of China Feng Liu, The Ministry-Province Jointly Constructed Base for State Key Lab-Shenzhen Key Laboratory of Chemical Biology, the Graduate School at Shenzhen, Tsinghua University, Shenzhen, 518055 People’s Republic of China Yu-Yang Jiang, Department of Chemistry, Tsinghua University, Beijing, 100084 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    We describe the development and characterization of a new cell line, designated Xl 1, derived from vertebra and long bones of Xenopus laevis . These cells can mineralize their extracellular matrix upon addition of an inorganic phosphate donor and vitamin C, as characterized by von Kossa staining. In addition they express genes such as matrix gla protein ( mgp ), alkaline phosphatase , type II collagen , and retinoic acid receptors , representing a valuable tool to analyze expression and regulation of Xenopus cartilage-associated genes. Continuous treatment with retinoic acid (RA) inhibited mineralization, alkaline phosphatase expression and its activity, suggesting that RA is a potential negative regulator of Xl 1 cell differentiation. These cells are receptive to efficient transfer of DNA using conventional methods including calcium phosphate, liposome-mediated transfer or electroporation and were found to express basal levels of mgp at least 50-fold higher than the routinely used Xenopus A6 cell line, as seen by transcription assays with the distal X. laevis mgp promoter. Being the first amphibian cell line derived from bone tissue, the Xl 1 culture provides an excellent in vitro tool for functional promoter studies, being suitable, among other uses, for identifying promoter elements mediating cartilage-expressed genes as shown here for mgp . Content Type Journal Article Pages 1-11 DOI 10.1007/s11010-012-1473-x Authors Natércia Conceição, Centre of Marine Sciences (CCMAR), University of Algarve, 8005-139 Faro, Portugal Michael Viegas, Centre of Marine Sciences (CCMAR), University of Algarve, 8005-139 Faro, Portugal João Fidalgo, Centre of Marine Sciences (CCMAR), University of Algarve, 8005-139 Faro, Portugal M. Leonor Cancela, Centre of Marine Sciences (CCMAR), University of Algarve, 8005-139 Faro, Portugal Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Insulin-like growth factor (IGF)-I and IGF-II play major roles in the regulation of skeletal muscle growth and differentiation, and both are locally expressed in muscle cells. Recent studies have demonstrated that IGF-II up-regulates its own gene expression during myogenesis and this auto-regulatory loop is critical for muscle differentiation. How local IGF-I is regulated in this process is unclear. Here, we report that while IGF-II up-regulated its own gene expression, it suppressed IGF-I gene expression during myogenesis. These opposite effects of IGF-II on IGF-I and IGF-II genes expression were time dependent and dose dependent. It has been shown that IGFs activate the PI3K-Akt-mTOR, p38 MAPK, and Erk1/2 MAPK pathways. In myoblasts, we examined their role(s) in mediating the opposite effects of IGF-II. Our results showed that both the PI3K-Akt-mTOR and p38 MAPK pathways played critical roles in increasing IGF-II mRNA expression. In contrast, mTOR was required for down-regulating the IGF-I gene expression by IGF-II. In addition, Akt, Erk1/2 MAPK, and p38 MAPK pathways were also involved in the regulation of basal levels of IGF-I and IGF-II genes during myogenesis. These findings reveal a previously unrecognized negative feedback mechanism and extend our knowledge of IGF-I and IGF-II gene expression and regulation during myogenesis. Content Type Journal Article Pages 1-7 DOI 10.1007/s11010-012-1479-4 Authors Shuang Jiao, Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003 People’s Republic of China Hongxia Ren, Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA Yun Li, Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003 People’s Republic of China Jianfeng Zhou, Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003 People’s Republic of China Cunming Duan, Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA Ling Lu, Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    To know the involvement of glycosaminoglycans (GAGs) in the metastasis of mouse FBJ osteosarcoma cells, N α -lauroyl- O -(β- d -xylopyranosyl)- l -serinamide (Xyl-Ser-C12), which initiates elongation of GAG chains using the glycan biosynthesis system in cells, was administered to FBJ cells with different metastatic capacities. Production of glycosylated products derived from Xyl-Ser-C12, especially heparan sulfate (HS) GAG-type oligosaccharides such as GalNAc-GlcA-GlcNAc-GlcA-Gal-Gal-Xyl-Ser-C12, was indicated in poorly metastatic FBJ-S1 cells more than in highly metastatic FBJ-LL cells by LC–MS. The results of RT-PCR revealed that HS synthases, Ext1 and Ext2, were expressed in FBJ-S1 cells more than in FBJ-LL cells. Furthermore, siRNA against Ext1 suppressed the expression of HS and enhanced the motility of FBJ-S1 cells. In addition, the expression of heparanase (HPSE) was enhanced in Ext-1-knockdown FBJ-S1 cells, and responsible for the increase in cell motility caused by the down-regulation of Ext1 expression. Our data provide the first evidence that Ext1 regulates the expression of HPSE and also indicated that levels of Ext1 and HPSE influenced the motility of FBJ cells. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1475-8 Authors Yinan Wang, Department of Biosciences and Informatics, Keio University, Hiyoshi, Yokohama, 223-8522 Japan XiaoYan Yang, Laboratory of Tumor Biology and Glycobiology, Department of Life Sciences and Biopharmaceutics, Shenyang Pharmaceutical University, P.O. Box 29, 103 WenHua Road, Shenyang, 110016 Liaoning, People’s Republic of China Sadako Yamagata, Laboratory of Tumor Biology and Glycobiology, Department of Life Sciences and Biopharmaceutics, Shenyang Pharmaceutical University, P.O. Box 29, 103 WenHua Road, Shenyang, 110016 Liaoning, People’s Republic of China Tatsuya Yamagata, Laboratory of Tumor Biology and Glycobiology, Department of Life Sciences and Biopharmaceutics, Shenyang Pharmaceutical University, P.O. Box 29, 103 WenHua Road, Shenyang, 110016 Liaoning, People’s Republic of China Toshinori Sato, Department of Biosciences and Informatics, Keio University, Hiyoshi, Yokohama, 223-8522 Japan Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    This study explored the role of pro- and anti-inflammatory cytokines in dimethyl benz(a)anthracene (DMBA)-induced lung cancer and its subsequent correction with a COX-2 inhibitory NSAID, etoricoxib. A single dose of DMBA (20 mg/kg body weight) in 0.9 % NaCl administered intratracheally was used to induce tumors in the rat lungs in 20 weeks. The study of pro-inflammatory cytokines like IL-1β, TNF-α, and IFN-γ revealed their upregulation by DMBA administration and restoration of their levels toward normal by the treatment with etoricoxib, while the anti-inflammatory cytokine IL-2 was found to be down-regulated with carcinogen administration and corrected with etoricoxib treatment. Apoptosis was studied by mitochondrial Bcl-2/Bax ratio and staining with fluorescent dyes acridine orange/ethidium bromide. The results showed a decreased apoptotic level with DMBA which was corrected with etoricoxib. Also, mitochondrial membrane potential was studied using JC-1 and rhodamine-123, which are membrane permeant fluorescent dyes, and generate information about cells at lower and higher mitochondrial membrane potential (∆Ψ M ). The results showed the presence of maximum number of cells with higher ∆Ψ M in the DMBA group and their number was considerably lowered in the other three groups. Content Type Journal Article Pages 1-12 DOI 10.1007/s11010-012-1451-3 Authors Neeti Nadda, Department of Biophysics, Panjab University, Chandigarh, 160014 India Shruti Setia, Department of Biophysics, Panjab University, Chandigarh, 160014 India Vivek Vaish, Department of Biophysics, Panjab University, Chandigarh, 160014 India Sankar Nath Sanyal, Department of Biophysics, Panjab University, Chandigarh, 160014 India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    With the increase of environment temperature, more and more attentions are payed to the effects of heat stress. Cells under heat shock either are adapted to the condition or are damaged and dead. In this paper, we found that heat shock induced endoplasmic reticulum (ER) stress. ATF4, PERK, and IRE1α were induced by heat shock of 45 °C in the transcriptional level. Under the stress of 45 °C, PERK was phosphorylated and XBP1s was detected. The result indicated that heat shock could induce the ER stress. We found that heat shock of 45 °C induced the dysregulation of HSP70 and DNA-PKcs, and downregulated the expression of PARP1 and XRCC1. Further results showed that after the knockdown of ATF4 or IRE1α, the expression of DNA-PKcs and XRCC1 were increased. It was indicated that ATF4 and IRE1α could inhibit the expression of DNA-PKcs and XRCC1 under the heat stress. Our results suggested that heat shock could activate ER stress. IRE1α and ATF4, as the important ER stress molecules, could inhibit the expression of DNA repair proteins DNA-PKcs, XRCC1, and HSP70 under heat shock. Downregulation of DNA repair proteins could aggravate the cell damage that may cause cell apoptosis. This may explain that heat shock could increase the lethality of chemotherapeutic drugs on tumor cells. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1439-z Authors Huifang Zhu, Department of Biochemistry and Molecular Biology, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing, 400016 China Feng-Jin Guo, Department of Cell Biology and Genetics, Core Facility of Development Biology, Chongqing Medical University, Chongqing, 400016 China Wenjun Zhao, Department of Cell Biology and Genetics, Core Facility of Development Biology, Chongqing Medical University, Chongqing, 400016 China Jinghua Zhou, Department of Cell Biology and Genetics, Core Facility of Development Biology, Chongqing Medical University, Chongqing, 400016 China Yanna Liu, Department of Cell Biology and Genetics, Core Facility of Development Biology, Chongqing Medical University, Chongqing, 400016 China Fangzhou Song, Department of Biochemistry and Molecular Biology, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing, 400016 China Yingxiong Wang, Laboratory of Reproductive Biology, Chongqing Medical University, Chongqing, 400016 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Placental growth factor (PlGF) inhibition produced promising results in reducing tumor burden in a diethylnitrosamine (DEN)-induced mouse model for hepatocellular carcinoma (HCC). The aim of this study was to non-invasively assess the improved histology by performing a serum glycomic analysis. To elucidate the molecular mechanism underlying the observed glycomic effects, we investigated the transcription and expression of E26 transformation-specific sequence 1 (Ets-1), a transcription factor essential for the glycomic and angiogenic changes in malignant transformation, including its different phosphorylated forms that result from activation of the MAP kinase and a Ca 2+ -dependent pathway. In addition, three Ets-1-dependent glycosyltransferase genes, Mgat4a , Mgat4b , and Mgat5 , were also evaluated. HCC was induced in mice by weekly injections with DEN for 16, 20, 25, and 30 w. In the treatment study, mice were injected with DEN for 25 w and subsequently treated with PlGF antibodies (5D11D4) for 5 w. Finally, PlGF−/− mice were injected with DEN for 20, 25, and 30 w. Serum N -glycans were analyzed with DNA sequencer-assisted fluorophore-assisted capillary electrophoresis and compared with histology. Maximum altered N -glycan phenotype was reached after 20 w of DEN-injections, i.e., when the first neoplastic lesions started to appear. 5D11D4-treatment improved the glycomic phenotype in that 7 of the 11 altered glycans tended to normalize. The PlGF−/− mice also showed a normalization trend, although not to the same extent of the treatment group. Number of Ets1 , Mgat4a , Mgat4b , and Mgat5 transcripts increased considerably in DEN-injected mice, however, a non-significant decrease was observed after 5D11D4-treatment. On the protein level, 5D11D4-treatment had a prominent effect on the MAP kinase pathway with a significant p38 activation, yet independent of Ets-1 function. Content Type Journal Article Pages 1-12 DOI 10.1007/s11010-012-1461-1 Authors Bram Blomme, Department of Hepatology and Gastroenterology, Ghent University Hospital, 9000 Ghent, Belgium Femke Heindryckx, Department of Hepatology and Gastroenterology, Ghent University Hospital, 9000 Ghent, Belgium Jean Marie Stassen, ThromboGenics NV, Leuven, Belgium Anja Geerts, Department of Hepatology and Gastroenterology, Ghent University Hospital, 9000 Ghent, Belgium Isabelle Colle, Department of Hepatology and Gastroenterology, Ghent University Hospital, 9000 Ghent, Belgium Hans Van Vlierberghe, Department of Hepatology and Gastroenterology, Ghent University Hospital, 9000 Ghent, Belgium Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    In the present study, the nephroprotective effect of gallic acid isolated from Peltiphyllum peltatum was examined in sodium fluoride (NaF) treated rats. Nephrotoxicity was induced by 1-week intoxication of NaF at 600 ppm through drinking water. The levels of thiobarbituric acid reactive substances, reduced glutathione as well as activities of superoxide dismutase and catalase in renal tissues homogenates were determined. The serum biochemical markers of renal injuries including creatinine, serum urea, blood urea nitrogen, uric acid levels as well as the levels of phosphate and calcium were also assessed. Intoxication with NaF caused a significant increase in the levels of thiobarbituric acid reactive substances (46 % versus to control) and reduced the glutathione concentration (47 %) and the activities of superoxide dismutase (46 %) and catalase (41 %) in renal tissues homogenates. NaF intoxication also induced significant alterations in the kidney biochemical markers increasing the levels of urea, uric acid, blood urea nitrogen, creatinine, and phosphate and decreasing the levels of calcium. Daily administration of gallic acid (20 mg/kg) for 1 week before NaF intoxication brought the antioxidant–oxidant balance similar to the NaF-untreated group. Silymarin, used a standard antioxidant agent, also showed a nephroprotective activity. We concluded that NaF caused nephrotoxicity and oxidative stress in renal tissues and daily administration of gallic acid for 1 week prior to intoxication inhibited toxicity and oxidative stress. Content Type Journal Article Pages 1-7 DOI 10.1007/s11010-012-1464-y Authors Seyed Mohammad Nabavi, Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran Solomon Habtemariam, Pharmacognosy Research Laboratories, Medway School of Science, University of Greenwich, Central Avenue, Chatham-Maritime, Kent, ME4 4TB UK Seyed Fazel Nabavi, Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran Antoni Sureda, Departament de Biologia Fonamental i Ciències de la Salut, Universitat de les Illes Balears, Illes Balears, Spain Maria Daglia, Department of Drug Sciences, School of Pharmacy, University of Pavia, Pavia, Italy Akbar Hajizadeh Moghaddam, Department of Biology, University of Mazandaran, Babolsar, Iran Mohammad Ali Amani, Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Schwann cells are critically important in recovery from injuries to the peripheral nervous system, and their absence from the central nervous system (CNS) may be a critical limiting factor in the CNS regeneration capacity. Various types of stem cells have been investigated for their potential to be induced to develop a Schwann cell phenotype, with mesenchymal stem cells (MSCs) being the most promising among them. The methods for inducing MSCs differentiation into Schwann cell-like cells are presented in detail in this review. The evidence related to successful differentiation of MSCs to Schwann cell-like cells is particularly discussed herein, which includes the changes in morphology, phenotype, function, and proteome. The possible explanations for the differentiation of MSCs to Schwann cell-like cells are also presented. Finally, we suggest future research aims which will need to be fulfilled to elucidate the biology of Schwann cell differentiation and MSC transdifferentiation, to enable clinical application of therapeutic differentiated MSC transplantation into nerve injury sites. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1351-6 Authors Yu Pan, Research Center of Medical Sciences, Guangdong General Hospital, Guangdong Academy of Medical Sciences, 96 Dongchuan Road, Guangzhou, 510080 China Sa Cai, Department of Physiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Integrins are transmembrane receptors that can specifically bind extracellular matrix (ECM) proteins. Assembly of the ECM protein fibronectin into fibrils has been shown to be a cell-mediated process that requires integrins. Like fibronectin, fibrillin 1 is an ECM glycoprotein that can assemble into fibrils, but the role of integrins in fibril formation is not understood. To investigate the role of integrins in fibrillin 1 ECM deposition, cells that normally produce and assemble fibrillin 1 fibers in vitro were stably transfected with plasmid constructs encoding short interfering RNAs that target specific integrin subunits. Cells that were deficient in α2- and β3-integrin subunits produced and deposited fibronectin normally, but cells that were deficient for α5 and αV were unable to elaborate a fibronectin matrix, although they continued to produce and secrete the protein. Surprisingly, the cells that were unable to elaborate a fibronectin matrix also lost fibrillin 1 gene expression. Content Type Journal Article Pages 1-12 DOI 10.1007/s11010-012-1383-y Authors Rajeev K. Boregowda, Division of Musculoskeletal Sciences, Department of Orthopaedics, Penn State College of Medicine, 500 University Drive, Hershey, PA 17033, USA Brooke M. Krovic, Division of Musculoskeletal Sciences, Department of Orthopaedics, Penn State College of Medicine, 500 University Drive, Hershey, PA 17033, USA Timothy M. Ritty, Division of Musculoskeletal Sciences, Department of Orthopaedics, Penn State College of Medicine, 500 University Drive, Hershey, PA 17033, USA Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Although the lipid mediator sphingosine 1-phosphate (S1P) has been identified to induce cell growth arrest of human keratinocytes, the sphingolipid effectively protects these epidermal cells from apoptosis. The molecular mechanism of the anti-apoptotic action induced by S1P is less characterized. Apart from S1P, endogenously produced nitric oxide (NO • ) has been recognized as a potent modulator of apoptosis in keratinocytes. Therefore, it was of great interest to elucidate whether S1P protects human keratinocytes via a NO • -dependent signalling pathway. Indeed, S1P induced an activation of endothelial nitric oxide synthase (eNOS) in human keratinocytes leading to an enhanced formation of NO • . Most interestingly, the cell protective effect of S1P was almost completely abolished in the presence of the eNOS inhibitor L-NAME as well as in eNOS-deficient keratinocytes indicating that the sphingolipid metabolite S1P protects human keratinocytes from apoptosis via eNOS activation and subsequent production of protective amounts of NO • . It is well established that most of the known actions of S1P are mediated by a family of five specific G protein-coupled receptors. Therefore, the involvement of S1P-receptor subtypes in S1P-mediated eNOS activation has been examined. Indeed, this study clearly shows that the S1P 3 is the exclusive receptor subtype in human keratinocytes which mediates eNOS activation and NO • formation in response to S1P. In congruence, when the S1P 3 receptor subtype is abrogated, S1P almost completely lost its ability to protect human keratinocytes from apoptosis. Content Type Journal Article Pages 1-12 DOI 10.1007/s11010-012-1433-5 Authors Elisabeth I. Schmitz, Institute of Nutritional Science, Toxicology, Faculty of Mathematics and Natural Science, University of Potsdam, Arthur-Scheunert-Allee, 114-116, 14558 Nuthetal, Germany Henrik Potteck, Institute of Pharmacy, Pharmacology and Toxicology, Freie Universität Berlin, Königin-Luise-Str. 2+4, 14195 Berlin, Germany Melanie Schüppel, Institute of Pharmacy, Pharmacology and Toxicology, Freie Universität Berlin, Königin-Luise-Str. 2+4, 14195 Berlin, Germany Marianti Manggau, Department of Pharmacology, Faculty of Pharmacy, Hasanuddin University, Jl. Perintis Kemerdekaan Km 10, Makassar, 90245 Indonesia Elly Wahydin, Department of Pharmacology, Faculty of Pharmacy, Hasanuddin University, Jl. Perintis Kemerdekaan Km 10, Makassar, 90245 Indonesia Burkhard Kleuser, Institute of Nutritional Science, Toxicology, Faculty of Mathematics and Natural Science, University of Potsdam, Arthur-Scheunert-Allee, 114-116, 14558 Nuthetal, Germany Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The present study was conducted to investigate if anti-inflammatory drug aspirin could alter the cytotoxic action of cisplatin on tumor cells. Using a transplantable T cell lymphoma in a murine model, we demonstrate that exposure to aspirin exerts a priming action on tumor cells, rendering them susceptible to induction of cell death by cisplatin with consequences on retardation of tumor progression. The priming action of aspirin on tumor cells was found to be dependent on an altered constitution of tumor microenvironment with respect to decline of acidosis and modulation in the expression of cell cycle and survival regulatory molecules like cyclin B1, cyclin D, bcl-2, bcl-xL, p53, and cytokines: IL-4, IL-10, IFN- γ & VEGF. The study also discusses possible mechanisms underlying augmentary action of aspirin on cisplatin-mediated tumor cells killing. This is the first report showing that pre-exposure of tumor cells to aspirin lowers the concentration of cisplatin to exert its cytotoxic action. The finding of this study will help in designing novel antitumor protocols with reduced dose of cisplatin. Content Type Journal Article Pages 1-12 DOI 10.1007/s11010-012-1421-9 Authors Anjani Kumar, School of Biotechnology, Banaras Hindu University, Varanasi, 221 005 Uttar Pradesh, India Sukh Mahendra Singh, School of Biotechnology, Banaras Hindu University, Varanasi, 221 005 Uttar Pradesh, India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Dendritic cells (DCs) are the most potent professional antigen-presenting cells and are involved in the initiation and progression of atherosclerosis. Recent data suggest that mature macrophages differentiate into dendritic-like cells when exposed to oxidized low-density lipoprotein (oxLDL). The purpose of the present study was to determine the effect of atorvastatin on the differentiation of macrophages to DCs and the molecular mechanisms of this transition. Mouse macrophage-like RAW264.7 cell was differentiated into a dendritic-like phenotype by incubation with oxLDL in the absence or presence of atorvastatin. The results showed that atorvastatin suppressed DC-like morphologic changes in vitro as assessed by decreased expression of DC maturation markers (CD83, CD11c, CD86, major histocompatibility complex class II, and CD1d). Atorvastatin also inhibited other oxLDL-induced functional changes including endocytic activity, ability to induce T cell proliferation, and cytokine secretion. Western blot analysis showed that oxLDL treatment of RAW264.7 cells induced phosphorylation of p38 mitogen-activated protein kinase (MAPK). However, blocking p38 MAPK with SB203580 significantly downregulated the expression of DC maturation markers, accompanied by decreased cytokine secretion. The findings of the present work demonstrate that that atorvastatin suppresses the oxLDL-induced DC-like differentiation of RAW264.7 cells by inactivating the p38 MAPK signaling pathway. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1427-3 Authors Liu-hua Hu, Department of Cardiology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200127 China Tuo Zhang, Department of Cardiology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200127 China Qin Shao, Department of Cardiology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200127 China Dan-dan Li, Department of Cardiology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200127 China Shu-xuan Jin, Department of Cardiology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200127 China Peng Nie, Department of Cardiology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200127 China Jing Yi, Department of Cell Biology, Key Laboratory of the Education Ministry for Cell Differentiation and Apoptosis, Institutes of Medical Sciences, Shanghai Jiaotong University School of Medicine, Shanghai, 200025 China Ben He, Department of Cardiology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200127 China Ling-hong Shen, Department of Cardiology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200127 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Oxidative stress is a major component of harmful cascades activated in neurodegenerative disorders. Here, we tried to elucidate the possible neuroprotective effect of Salvigenin, a natural polyphenolic compound, on oxidative stress-induced apoptosis and autophagy in human neuroblastoma SH-SY5Y cells. We measured cell viability by MTT test and found that 25 μM is the best protective concentration of Salvigenin. GSH and SOD assays suggested that Salvigenin activates antioxidant factors. At the same time, measurement of ER stress-associated proteins including calpain and caspase-12 showed the ability of Salvigenin to decrease ER stress. We found that Salvigenin could decrease the apoptotic factors. Salvigenin inhibited H 2 O 2 -induced caspase-3 which is a hallmark of apoptosis in addition to reducing Bax\Bcl-2 ratio by 1.45 fold. Additionally, Salvigenin increased the levels of autophagic factors. Our results showed an increase in LC3-II/LC3-I ratio, Atg7, and Atg12 in the presence of 25 μM of Salvigenin by about 1.28, 1.25, and 1.54 folds, respectively, compared to H 2 O 2 -treated cells. So it seems that H 2 O 2 cytotoxicity mainly results from apoptosis. Besides, Salvigenin helps cells to survive by inhibiting apoptosis and enhancing autophagy that opens a new horizon for the future experiments. Content Type Journal Article Pages 1-14 DOI 10.1007/s11010-012-1416-6 Authors Ghazaleh Rafatian, Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran Fariba Khodagholi, Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran Mahdi Moridi Farimani, Department of Phytochemistry, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, G. C., Evin, Tehran, Iran Shahnaz Babaei Abraki, Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran Mossa Gardaneh, Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Apigenin is a naturally occurring plant flavone with strong anti-oxidant and anti-inflammatory activity. While the anticancer properties of Apigenin have been extensively studied, little is known about its effects on endothelial dysfunction. We investigated the effects of Apigenin in EAhy926 endothelial cells exposed to TNFα by evaluating the expression of eNOS and MMP-9, two key molecules in endothelial dysfunction. MMP-9 activity was measured by gel zymography. Western blot analysis was performed to analyze eNOS expression and signal transduction. Treatment with Apigenin (50 μM) counteracted the TNFα-induced expression of eNOS and MMP-9 and the TNFα- triggered activation of Akt, p38MAPK and JNK signalling suggesting that multiple signalling pathways are involved in mediating the protective effects of Apigenin on endothelial function. To better understand the molecular mechanisms underlying the protective effects of Apigenin, we used a pharmacological approach with specific inhibitors. The use of an Akt inhibitor mimicked the inhibitory effects of Apigenin on eNOS and MMP-9 expression, suggesting that eNOS and MMP-9 induction by TNFα depends on Akt activation. The TNFα-induced expression of MMP-9 was also affected by the JNK inhibitor SP600125. No effect on eNOS and MMP-9 expression was observed in the presence of the p38MAPK inhibitor SB203580 or the ERK 1/2 inhibitor PD98059. Pretreatment with ‘classic’ (ERα and ERβ) or ‘non classic’ (GPR30) oestrogen receptor (ER) inhibitors (ICI182,780 and PTX, respectively) counteracted the ability of Apigenin to decrease the TNFα-triggered activation of the Akt pathway. Consistently, the use of both ER inhibitors reversed the inhibitory effects of Apigenin on the TNFα-induced expression of eNOS and, to a lesser extent, MMP-9. We can conclude that Apigenin exerts its inhibitory effect on the TNFα-induced expression of eNOS and MMP-9 through the Akt signalling inhibition generated by ER activation. Oestrogen signalling has been implicated in protection from cardiovascular disease. Therefore, having regard to its ability to bind to ERs, Apigenin may be considered an oestrogen-like molecule to potentially be used against the onset and progression of vascular diseases associated with endothelial dysfunction. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1429-1 Authors Daniela Palmieri, Vascular and Endovascular Unit, Laboratory of Clinical and Experimental Vascular Biology, DISC, University Hospital IRCCS San Martino, Largo Rosanna Benzi 8, 16132 Genoa, Italy Patrizia Perego, Department of Chemical and Process Engineering “G.B. Bonino”, University of Genoa, Via Opera Pia 15, 16145 Genoa, Italy Domenico Palombo, Vascular and Endovascular Unit, Laboratory of Clinical and Experimental Vascular Biology, DISC, University Hospital IRCCS San Martino, Largo Rosanna Benzi 8, 16132 Genoa, Italy Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Human atrial and ventricular myocardium has distinct structure and physiology. MicroRNAs (miRNAs) are the central players in the regulation of gene expression, participating in many physiological processes. A comprehensive knowledge of miRNA expression in the human heart is essential for the understanding of myocardial function. The aim of this study was to compare the miRNA signature in human right atrial and ventricular myocardium. Agilent human miRNA arrays were used to indicate the miRNA expression signatures of the right atrial ( n  = 8) and ventricular ( n  = 9) myocardium of healthy individuals. Quantitative reverse transcription-polymerase chain reactions (qRT-PCRs) were used to validate the array results. DIANA-mirPath was used to incorporate the miRNAs into pathways. MiRNA arrays showed that 169 miRNAs were expressed at different levels in human right atrial and ventricular myocardium. The unsupervised hierarchical clustering analysis based on the 169 dysregulated miRNAs showed that miRNA expression categorized two well-defined clusters that corresponded to human right atrial and ventricular myocardium. The qRT-PCR results correlated well with the microarray data. Bioinformatic analysis indicated the potential miRNA targets and molecular pathways. This study indicates that distinct miRNA expression signatures in human right atrial and ventricular myocardium. The findings provide a novel understanding of the molecular differences between human atrial and ventricular myocardium and may establish a framework for an anatomically detailed evaluation of cardiac function regulation. Content Type Journal Article Pages 1-7 DOI 10.1007/s11010-012-1417-5 Authors Yangyang Zhang, Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029 People’s Republic of China Xiaowei Wang, Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029 People’s Republic of China Xiaohan Xu, Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029 People’s Republic of China Jun Wang, Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029 People’s Republic of China Xiang Liu, Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029 People’s Republic of China Yijiang Chen, Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The lactate dehydrogenase (LDH) has been studied widely because it exists in various isozymic forms. The association of A and B subunits of LDH can generate five tetrameric isozymes, but the finding of the sixth isozyme in mature human testis and sperm indicated the presence of an additional subunit of LDH, designated as LDH-X (also termed LDH-C4 due to tetrameric nature of C-subunit). LDH-C4 isozyme is an iso-, allo-, and auto-antigen present in mammalian sperm cells. The synthesis of LDH-C4 in the testis takes place during sexual maturation, and it is the predominant fraction in mature spermatozoa. Though, originally considered to be testis specific, LDH-C or Ldh3 in mice was later detected in the murine oocyte and early embryo. Ldh3 in mouse supports its role in energy production in spermatids that favor lactate as substrate and in spermatozoa with a characteristic aerobic glycolytic path to yield ATP. During last two decades, cancer/testis-associated genes (CTAs) which are expressed only in the germinal epithelium of the testis are also expressed in some cancer cells, but not in non-cancerous somatic tissues. The CTAs are considered promising candidates for diagnosis and immunotherapy of cancer. The sperm-specific Ldh - c gene has been shown to express in a broad spectrum of human tumors, with high frequency in lung cancer, melanoma, and breast cancer; the protein being expressed virtually in all tumor types tested. Accordingly, LDH-C4 is the unique target for contraception in both males and females and offers potential future for immunotherapy of different types of cancers. As LDH-C has a preference for lactate as a substrate, LDH-C activation in cancer may depend on lactate for ATP production. The major aim of this article is to review the salient features of LDH-C subunit and the immune responses of LDH-C4 in homologous and heterologous species in relation to its role in acceptance or rejection of the allograft and its application in contraception and immunotherapy of cancer, directly or indirectly through the regulation of its substrate, the lactate. Content Type Journal Article Pages 1-13 DOI 10.1007/s11010-012-1428-2 Authors G. S. Gupta, Department of Biophysics, Panjab University, Chandigarh, 160014 India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Our aim was to examine correlations between polymorphisms in five antioxidant enzymes genes, activity of free-radical processes, and the risk of restenosis after coronary artery stenting with bare metal stents (BMS). A total of 101 male patients who underwent intracoronary stenting using BMS and coronary angiography follow-up of 6 months were enrolled in: group with in-stent restenosis ( n  = 44) and without restenosis ( n  = 57). The content of lipoperoxides and malondialdehyde (MDA) in Low-density lipoprotein (LDL), activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) in erythrocytes, and genotypes polymorphisms of the CAT gene (−262C/T), paraoxonase-1 ( PON - 1 ) gene (163T/A and 575A/G), endothelial nitric oxide synthase ( eNOS ) gene (298G/T (rs#1799983) and −786T/C), GPx - 1 gene (599C/T (rs#1050450)), and glutathione-S-transferase ( GSTP ) gene (313A/G) were determined. In carriers of the minor allele of 599C/T polymorphism of the GPx - 1 gene, activity of GPx in erythrocytes was lower by 17 % than in wild allele homozygotes, while the content of lipoperoxides in LDL was higher by 74 %. T-allele of 599C/T polymorphism of the GPx - 1 gene (OR = 2.9; 95 % CI: 1.23–6.82) and T-allele of 298G/T polymorphism of the eNOS gene (OR = 2.79; 95 % CI: 1.17–6.66) were associated with the risk of in-stent restenosis. Minor alleles of polymorphisms 298G/T of the eNOS gene and 599C/T of the GPx - 1 gene are associated with an increased risk of in-stent restenosis. Minor allele of the GPx - 1 gene 599C/T polymorphism leads to a decrease of the GPx activity and increase of the activity of free-radical processes. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1419-3 Authors Yu. A. Shuvalova, Department of Atherosclerosis Problems, Russian Cardiology Research and Production Complex of the Russian Federation Ministry of Health and Social Development, 3rd Cherepkovskaya Street, 15a, Moscow, 121552 Russia A. I. Kaminnyi, Department of Atherosclerosis Problems, Russian Cardiology Research and Production Complex of the Russian Federation Ministry of Health and Social Development, 3rd Cherepkovskaya Street, 15a, Moscow, 121552 Russia A. N. Meshkov, Department of Clinical Cardiology and Molecular Genetics, National Center of Preventive Medcine, Moscow, Russia R. O. Shirokov, Laboratory of Radio Vascular Therapy, Russian Cardiology Research and Production Complex of the Russian Federation Ministry of Health and Social Development, Moscow, Russia A. N. Samko, Laboratory of Radio Vascular Therapy, Russian Cardiology Research and Production Complex of the Russian Federation Ministry of Health and Social Development, Moscow, Russia Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    High-density lipoprotein (HDL) possesses protective properties in cardiovascular diseases. However, the effect of HDL on the mesenchymal stem cells (MSCs), which could be mobilized to the damaged myocardial tissue, has not been well elucidated yet. In the current study, we investigated the effect of HDL on the proliferation of MSCs so as to reveal its molecular mechanisms. MSCs derived from rats were treated with HDL in different concentrations and for different periods. The proliferation of MSCs was measured with MTT and BrdU cell proliferation assay. The phosphorylation of Akt, ERK1/2 and the expression of p21 were evaluated by Western blotting. After the activity of respective pathways was down-regulated by the specific inhibitor and the gene of scavenger receptor-B type I (SR-BI) was knocked down by RNA interference, BrdU assay was performed to examine this effect of HDL on MSCs. We found that the proliferation of MSCs induced by HDL, in a time- and concentration-dependent manner, was the phosphorylation of Akt- and ERK1/2-dependent, which was significantly attenuated by the specific inhibitor to respective pathways. Moreover, MAPK/ERK1/2 pathway exerted a more dominating effect on this process. SR-BI contributed to HDL-induced proliferation of MSCs, which was effectively abolished by the silencing of SR-BI. The results suggested that HDL was capable of improving MSCs proliferation, in which MAPK/ERK1/2 and PI3K/Akt pathways involved and SR-BI played a critical role as well. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1422-8 Authors Jianfeng Xu, Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, 200032 China Juying Qian, Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, 200032 China Xinxing Xie, Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, 200032 China Li Lin, Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, 200032 China Jianying Ma, Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, 200032 China Zheyong Huang, Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, 200032 China Mingqiang Fu, Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, 200032 China Yunzeng Zou, Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, 200032 China Junbo Ge, Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, 200032 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    High levels of homocysteine (Hcy), known as hyperhomocysteinemia (HHcy), are associated with cerebrovascular diseases, such as vascular dementia, stroke, and Alzheimer’s disease. The γ-amino butyric acid (GABA) is an inhibitory neurotransmitter and a ligand of GABA-A receptor. By inhibiting excitatory response, it may decrease complications associated with vascular dementia and stroke. Hcy specifically competes with the GABA-A receptors and acts as an excitotoxic neurotransmitter. Previously, we have shown that Hcy increases levels of NADPH oxidase and reactive oxygen species (ROS), and decreases levels of thioredoxin and peroxiredoxin by antagonizing the GABA-A receptor. Hcy treatment leads to activation of matrix metalloproteinases (MMPs) in cerebral circulation by inducing redox stress and ROS. The hypothesis is that Hcy induces MMPs and suppresses tissue inhibitors of metalloproteinase (TIMPs), in part, by inhibiting the GABA-A receptor. This leads to degradation of the matrix and disruption of the blood brain barrier. The brain cortex of transgenic mouse model of HHcy (cystathionine β-synthase, CBS−/+) and GABA-A receptor null mice treated with and without muscimol (GABA-A receptor agonist) was analysed. The mRNA levels were measured by Q-RT-PCR. Levels of MMP-2, -9, -13, and TIMP-1, -2, -3, and -4 were evaluated by in situ labeling and PCR-gene arrays. Pial venular permeability to fluorescence-labeled albumin was assessed with intravital fluorescence microscopy. We found that Hcy increases metalloproteinase activity and decreases TIMP-4 by antagonizing the GABA-A receptor. The results demonstrate a novel mechanism in which brain microvascular permeability changes during HHcy and vascular dementias, and have therapeutic ramifications for microvascular disease in Alzheimer’s patients. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1425-5 Authors David Lominadze, Department of Physiology and Biophysics, University of Louisville School of Medicine, Bldg. A, Room 1115, 500 South Preston Street, Louisville, KY, 40202, USA Neetu Tyagi, Department of Physiology and Biophysics, University of Louisville School of Medicine, Bldg. A, Room 1115, 500 South Preston Street, Louisville, KY, 40202, USA Utpal Sen, Department of Physiology and Biophysics, University of Louisville School of Medicine, Bldg. A, Room 1115, 500 South Preston Street, Louisville, KY, 40202, USA Alexander Ovechkin, Department of Physiology and Biophysics, University of Louisville School of Medicine, Bldg. A, Room 1115, 500 South Preston Street, Louisville, KY, 40202, USA Suresh C. Tyagi, Department of Physiology and Biophysics, University of Louisville School of Medicine, Bldg. A, Room 1115, 500 South Preston Street, Louisville, KY, 40202, USA Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The human papilloma virus E4 protein is highly expressed in late times of infection. Evidence to date suggests that E4 is essential for amplification of the viral genome and that it can influence cell cycle. Examination of the sequences encoding the E4 proteins from several genotypes of human papillomavirus revealed the presence of RXL-containing motifs reminiscent of the cyclin-binding motifs that have been identified in several cyclin-binding proteins. When baculovirus-produced human cyclin E and cyclin A with cdk2 were incubated in vitro with a GST-E4 fusion protein, both cyclin E and A stably interacted with the GST-E4 protein containing the full E4 sequence from HPV18. The interaction was not dependent on the presence of the kinase subunit but was dependent on the integrity of the RXL motif in E4. When incubated with cell extracts from the C33A human cervical carcinoma cell line or when expressed in C33A cells, the GST-E4 protein formed interactions with cyclin A and cdk2 and kinase activity could be demonstrated in the GST-E4 complex. In contrast to the baculovirus-produced cyclin E, cellular cyclin E failed to detectably interact with GST-E4 suggesting that the HPV18 E4 sequences are capable of interacting only with cyclin A in mammalian cells. These observations suggest that human papillomavirus E4 proteins can interact with cyclin A/cdk2, which may contribute to viral manipulation of the host cell cycle. Content Type Journal Article Pages 1-12 DOI 10.1007/s11010-012-1472-y Authors Qingming Ding, Department of Pathology and Molecular Medicine, McMaster University, 1280 Main Street West, Hamilton, ON L8S 4K1, Canada Lili Li, Department of Pathology and Molecular Medicine, McMaster University, 1280 Main Street West, Hamilton, ON L8S 4K1, Canada Peter Whyte, Department of Pathology and Molecular Medicine, McMaster University, 1280 Main Street West, Hamilton, ON L8S 4K1, Canada Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Atherosclerosis is one of the major complications of diabetes and involves endothelial dysfunction, matrix alteration, and most importantly migration and proliferation of vascular smooth muscle cells (VSMCs). Although hyperglycemia and hyperinsulinemia are known to contribute to atherosclerosis, little is known about the specific cellular signaling pathways that mediate the detrimental hyperinsulinemic effects in VSMCs. Therefore, we investigated the cellular mechanisms of hyperinsulinemia-induced migration and proliferation of VSMCs. VSMCs were treated with insulin (100 nM) for 6 days and subjected to various physiological and molecular investigations. VSMCs subjected to hyperinsulinemia exhibited increased migration and proliferation, and this is paralleled by oxidative stress [increased NADPH oxidase activity, NADPH oxidase 1 mRNA expression, and reactive oxygen species (ROS) generation], alterations in mitochondrial physiology (membrane depolarization, decreased mitochondrial mass, and increased mitochondrial ROS), changes in mitochondrial biogenesis-related genes (mitofusin 1, mitofusin 2, dynamin-related protein 1, peroxisome proliferator-activated receptor gamma coactivator 1-alpha, peroxisome proliferator-activated receptor gamma coactivator 1-beta, nuclear respiratory factor 1, and uncoupling protein 2), and increased Akt phosphorylation. Diphenyleneiodonium, a known NADPH oxidase inhibitor significantly inhibited migration and proliferation of VSMCs and normalized all the above physiological and molecular perturbations. This study suggests a plausible crosstalk between mitochondrial dysfunction and oxidative stress under hyperinsulinemia and emphasizes counteracting mitochondrial dysfunction and oxidative stress as a novel therapeutic strategy for atherosclerosis. Content Type Journal Article Pages 1-11 DOI 10.1007/s11010-012-1478-5 Authors Shiny Abhijit, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, Gopalapuram, Chennai, 600 086 India Regin Bhaskaran, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, Gopalapuram, Chennai, 600 086 India Abirami Narayanasamy, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, Gopalapuram, Chennai, 600 086 India Anand Chakroborty, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, Gopalapuram, Chennai, 600 086 India Nagaraj Manickam, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, Gopalapuram, Chennai, 600 086 India Madhulika Dixit, Department of Biotechnology, Indian Institute of Technology, Chennai, India Viswanathan Mohan, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, Gopalapuram, Chennai, 600 086 India Muthuswamy Balasubramanyam, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, Gopalapuram, Chennai, 600 086 India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Quercetin has been reported to protect testicular cells from oxidative damage induced by environmental chemicals. In this study, we isolated interstitial Leydig cells (ILCs) from immature rats, set-up ILCs culture, co-treated cells with atrazine (ATZ) and quercetin (QT), evaluated toxicity, and measured the expression levels of antioxidant enzymes and nuclear factor-kappaB (NF-κB) and levels of steroidogenic enzymes. ATZ decreased ILCs viability at concentrations higher than 10 μg/mL and increased reactive oxygen species, malondialdehyde (MDA), and glutathione levels. ATZ also increased glutathione peroxidase, glutathione reductase, and glutathione-S-transferase and decreased superoxide dismutase-1 (s od1 ) and superoxide dismutase-2 (s od2 ) messenger RNA (mRNA) levels which were prevented by QT. The changes in the MDA levels and lactate dehydrogenase leakage induced by ATZ (50 μg/mL) were also prevented on co-treatment with QT (50 μM). Furthermore, ATZ-induced 3β- and 17β-hydroxysteroid dehydrogenase activities and NF-κB-expressions at the mRNA and protein levels were also recovered to control value on co-treatment with QT. These data showed that QT protected against ATZ-induced ILCs toxicity by restoring the expression of NF-κB and steroidogenic activity and by preventing the oxidative stress. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1471-z Authors Sunny O. Abarikwu, Department of Chemical Sciences, College of Natural Sciences, Redeemer’s University, Redemption City, Ogun State, Nigeria Aditya B. Pant, Indian Institute of Toxicology Research (Formerly Industrial Toxicology Research Centre), Lucknow, India Ebenezer O. Farombi, Drug Metabolism and Toxicology Research laboratories, Department of Biochemistry, University of Ibadan, Ibadan, Oyo State, Nigeria Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Thrombin and platelet-derived growth factor-B chain (PDGF-B) are key factors in the stimulation of atherosclerosis. The effect of thrombin on PDGF-B production has been characterized. However, the underlying mechanism is still far clear. Here, we investigate the transcription factors and regulators that are involved in PDGF-B production caused by thrombin in endothelial cells (ECs). Levels of PDGF were analyzed by real-time RT-PCR and ELISA, while levels of early growth response-1 (Egr-1) were analyzed by real-time RT-PCR and western blot. To evaluate the function of CBP and Egr-1 involved in regulation of PDGF-B, small interfering RNA (siRNA) were used to down-regulate their expression in mRNA and protein level. Interaction of Egr-1 and CBP was measured with immunoprecipitation and western blot. Thrombin induced an early and transient up-regulation of transcription factor early Egr-1, which was followed by a delayed increase of PDGF-B. siRNA against Egr-1-inhibited thrombin-induced PDGF-B production. Furthermore, thrombin could enhance the interaction of Egr-1 with its co-activator CREB-binding protein (CBP). CBP knockdown attenuated this interaction, and led to a reduction of PDGF-B expression induced by thrombin. Our results suggest that CBP might be one of the main interaction targets for Egr-1, and the transient activation of Egr-1 and recruitment of CBP are required for thrombin-induced PDGF-B in ECs. Content Type Journal Article Pages 1-7 DOI 10.1007/s11010-012-1285-z Authors Jing Chen, Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan, 430060 China Lin Xu, Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan, 430060 China Sisi Chen, Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan, 430060 China Jian Yang, Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan, 430060 China Hong Jiang, Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan, 430060 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    We have previously reported Foxk1 as an important transcription factor in the myogenic progenitors. SWI-independent-3 (Sin3) has been identified as a Foxk1 binding candidate using a yeast two-hybrid screen. In the present study, we have identified the Foxk1 N-terminal (1–40) region as the Sin3 interacting domain (SID), and the PAH2 of Sin3 as the Foxk1 binding domain utilizing yeast two-hybrid and GST pull-down assays. Further studies revealed that knockdown of Sin3a or Sin3b results in cell cycle arrest and upregulation of cell cycle inhibitor genes. In summary, our present studies have shown that Foxk1 interacts with Sin3 through the SID and that Sin3 has an important role in the regulation of cell cycle kinetics of the MPC population. The results of these studies continue to define and assemble the networks that regulate the MPCs and muscle regeneration. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1302-2 Authors Xiaozhong Shi, Lillehei Heart Institute, University of Minnesota-Twin Cities, 4-108 NHH, 312 Church St SE, Minneapolis, MN 55455, USA Daniel J. Garry, Lillehei Heart Institute, University of Minnesota-Twin Cities, 4-108 NHH, 312 Church St SE, Minneapolis, MN 55455, USA Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Previous studies suggested that activated c-Src promote the tyrosine phosphorylation of NMDA receptor subunit NR2A, and thus aggravate the injury induced by transient cerebral ischemia/reperfusion (I/R) in rat hippocampus CA1 region. In this study, we examined the effect of nitric oxide (NO) on the activation of c-Src and the tyrosine phosphorylation of NMDA receptor NR2A subunit. The results show that S-nitrosylation and the phosphorylation of c-Src were induced after cerebral I/R in rats, and administration of nNOS inhibitor 7-NI, nNOS antisense oligonucleotides and exogenous NO donor sodium nitroprusside diminished the increased S-nitrosylation and phosphorylation of c-Src during cerebral I/R. The cysteine residues of c-Src modified by S-nitrosylation are Cys489, Cys498, and Cys500. On the other hand, NMDAR antagonist MK-801 could attenuate the S-nitrosylation and activation of c-Src. Taken together, the S-nitrosylation of c-Src is provoked by NO derived from endogenous nNOS, which is activated by Ca 2+ influx from NMDA receptors, and promotes the auto-phosphorylation at tyrosines and further phosphorylates NR2A. The molecular mechanism we outlined here is a novel postsynaptic NMDAR-nNOS/c-Src-mediated signaling amplification, the ‘NMDAR-nNOS → NO → SNO-c-Src → p-c-Src → NMDAR-nNOS’ cycle, which presents the possibility as a potential therapeutic target for stroke treatment. Content Type Journal Article Pages 1-15 DOI 10.1007/s11010-012-1280-4 Authors Li-Juan Tang, Research Center of Biochemistry and Molecular Biology and Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical College, 84 West Huai-hai Road, Xuzhou, 221002 Jiangsu, People’s Republic of China Chong Li, Research Center of Biochemistry and Molecular Biology and Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical College, 84 West Huai-hai Road, Xuzhou, 221002 Jiangsu, People’s Republic of China Shu-Qun Hu, Research Center of Biochemistry and Molecular Biology and Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical College, 84 West Huai-hai Road, Xuzhou, 221002 Jiangsu, People’s Republic of China Yong-Ping Wu, Jiangsu Key Laboratory of Anesthesiology, Xuzhou Medical College, Xuzhou, Jiangsu, People’s Republic of China Yan-Yan Zong, Research Center of Biochemistry and Molecular Biology and Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical College, 84 West Huai-hai Road, Xuzhou, 221002 Jiangsu, People’s Republic of China Chang-Cheng Sun, Research Center of Biochemistry and Molecular Biology and Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical College, 84 West Huai-hai Road, Xuzhou, 221002 Jiangsu, People’s Republic of China Fa Zhang, Research Center of Biochemistry and Molecular Biology and Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical College, 84 West Huai-hai Road, Xuzhou, 221002 Jiangsu, People’s Republic of China Guang-Yi Zhang, Research Center of Biochemistry and Molecular Biology and Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical College, 84 West Huai-hai Road, Xuzhou, 221002 Jiangsu, People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Identifying low-abundance mutations is important for the therapy and diagnose of cancer. Since the potential for tumor heterogeneity, the efficient detection of cancer-relevant mutations largely depends on the sensitivity of the methods employed. To confirm whether the mutation detection platforms affect the perceived prevalence of the BRAF V600E and its correlation with clinicopathologic features in papillary thyroid carcinomas (PTC), we compared Sanger Sequencing (SS), Pyrosequencing (PS), and a newly built allele-specific real-time PCR (AS-qPCR) apparatus for the detection of BRAF V600E in a Chinese cohort of conventional variant PTC. Accurate plasmid standards were built to assess the limit of detection of the three platforms. In this research, AS-qPCR has been found both the most sensitive and reliable at detecting mutation. The mutations detected by AS-qPCR which were not detected by SS or PS due to low abundance were confirmed by mutation enrichment platform COLD-PCR followed by SS. When analyzed by AS-qPCR, BRAF V600E was associated with a more aggressive phenotype. Our results indicate that the reported prevalence of the BRAF V600E mutations in PTC has been underestimated and more sensitive methods such as AS-qPCR should be applied in clinical settings. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1282-2 Authors Weiqin Jiang, Cancer Center, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003 China Weibin Wang, Cancer Center, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003 China FangFang Fu, Institute of Infectious Diseases, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003 China Xiaodong Teng, Department of Pathology, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003 China Haohao Wang, Cancer Center, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003 China Haiyong Wang, Cancer Center, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003 China Lisong Teng, Cancer Center, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Although shortened telomeres were shown associated with several risk factors of diabetes, there is lack of data on their relationship with mitochondrial dysfunction. Therefore, we compared the relationship between telomere length and mitochondrial DNA (mtDNA) content in patients with type 2 diabetes mellitus (T2DM; n  = 145) and in subjects with normal glucose tolerance (NGT; n  = 145). Subjects were randomly recruited from the Chennai Urban Rural Epidemiology Study. mtDNA content and telomere length were assessed by Real-Time PCR. Malonodialdehyde, a marker of lipid peroxidation was measured by thiobarbituric acid reactive substances (TBARS) using fluorescence methodology. Adiponectin levels were measured by radioimmunoassay. Oxidative stress as determined by lipid peroxidation (TBARS) was significantly ( p  〈 0.001) higher in patients with T2DM compared to NGT subjects. In contrast, the mean telomere length, adiponectin and mtDNA content were significantly ( p  〈 0.001) lower in patients with T2DM compared to NGT subjects. Telomere length was positively correlated with adiponectin, HDL, mtDNA content and good glycemic/lipid control and negatively correlated with adiposity and insulin resistance. On regression analysis, shortened telomeres showed significant association with T2DM even after adjusting for waist circumference, insulin resistance, triglyceride, HDL, adiponectin, mtDNA & TBARS. mtDNA depletion showed significant association with T2DM after adjusting for waist circumference and adiponectin but lost its significance when further adjusted for telomere length, TBARS and insulin resistance. Our study emphasizes the clustering of accelerated aging features viz., shortened telomeres, decreased mtDNA content, hypoadiponectinemia, low HDL, and increased oxidative stress in Asian Indian type 2 diabetes patients. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1276-0 Authors Finny Monickaraj, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, WHO Collaborating Centre for Non-Communicable Diseases Prevention and Control, IDF Centre of Education, Gopalapuram, Chennai, 600 086 Tamilnadu, India Sankaramoorthy Aravind, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, WHO Collaborating Centre for Non-Communicable Diseases Prevention and Control, IDF Centre of Education, Gopalapuram, Chennai, 600 086 Tamilnadu, India Kuppan Gokulakrishnan, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, WHO Collaborating Centre for Non-Communicable Diseases Prevention and Control, IDF Centre of Education, Gopalapuram, Chennai, 600 086 Tamilnadu, India Chandrakumar Sathishkumar, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, WHO Collaborating Centre for Non-Communicable Diseases Prevention and Control, IDF Centre of Education, Gopalapuram, Chennai, 600 086 Tamilnadu, India Paramasivam Prabu, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, WHO Collaborating Centre for Non-Communicable Diseases Prevention and Control, IDF Centre of Education, Gopalapuram, Chennai, 600 086 Tamilnadu, India Durai Prabu, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, WHO Collaborating Centre for Non-Communicable Diseases Prevention and Control, IDF Centre of Education, Gopalapuram, Chennai, 600 086 Tamilnadu, India Viswanathan Mohan, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, WHO Collaborating Centre for Non-Communicable Diseases Prevention and Control, IDF Centre of Education, Gopalapuram, Chennai, 600 086 Tamilnadu, India Muthuswamy Balasubramanyam, Department of Cell and Molecular Biology, Madras Diabetes Research Foundation and Dr. Mohan’s Diabetes Specialities Centre, WHO Collaborating Centre for Non-Communicable Diseases Prevention and Control, IDF Centre of Education, Gopalapuram, Chennai, 600 086 Tamilnadu, India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Patients with type 2 diabetes (T2D) manifest significant abnormalities in lipoprotein structure and function. The deleterious impact of oxidative and glycoxidative modifications on HDL-mediated atheroprotective, antiinflammatory, and antioxidative phenomena has been well established. However, the biological effects of modified HDL on adrenal steroidogenesis—which could reveal a pathophysiological link to the overactivity of the renin–angiotensin–aldosterone system and its adverse cardiovascular consequences often observed in T2D—are not well delineated. We studied the role of modified HDL on aldosterone release from adrenocortical carcinoma cells (NCI-H295R). In vitro modifications of native HDL were performed in the presence of glucose for glycoxidized HDL (glycoxHDL) and sodium hypochlorite for oxidized HDL. Angiotensin II (AngII)-sensitized H295R cells were treated with lipoproteins for 24 h, and supernatant was used to measure aldosterone release. Both native and modified HDL augmented the steroid release from AngII-sensitized cells, with glycoxHDL having the greatest impact. Both the modified forms of HDL induced a significant increase in scavenger receptor expression and employed protein kinase C as well as extracellular signal-regulated kinase as downstream effectors of aldosterone release. Native HDL and modified HDL required Janus kinase-2 for combating increased demand in steroidogenesis. Therefore, our data support the hypothesis that diabetes-induced modification of HDL may promote adrenocortical aldosterone secretion via different signal transduction pathways. This significant influence on multiple signaling mechanisms could be targeted for future research to implement novel therapeutic trials. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1274-2 Authors Sarama Saha, Department of Internal Medicine III, Carl Gustav Carus Medical School, Technical University of Dresden, Fetscherstraße 74, 01307 Dresden, Germany Juergen Graessler, Department of Internal Medicine III, Carl Gustav Carus Medical School, Technical University of Dresden, Fetscherstraße 74, 01307 Dresden, Germany Peter E. H. Schwarz, Department of Internal Medicine III, Carl Gustav Carus Medical School, Technical University of Dresden, Fetscherstraße 74, 01307 Dresden, Germany Claudia Goettsch, Department of Internal Medicine III, Carl Gustav Carus Medical School, Technical University of Dresden, Fetscherstraße 74, 01307 Dresden, Germany Stefan R. Bornstein, Department of Internal Medicine III, Carl Gustav Carus Medical School, Technical University of Dresden, Fetscherstraße 74, 01307 Dresden, Germany Steffi Kopprasch, Department of Internal Medicine III, Carl Gustav Carus Medical School, Technical University of Dresden, Fetscherstraße 74, 01307 Dresden, Germany Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Tyrosinase is the key regulatory enzyme of melanogenesis and plays a major role in mammal coat color. For the first time, we have sequenced and characterized the tyrosinase (TYR) of Jining Gray Goat ( Capra hircus ), which is the world-famous fur-bearing animal with its special color and pattern. The full-length cDNA was cloned by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end (RACE) method. As a result, one 2131-bp nucleotide sequence representing the full-length cDNA of TYR was obtained. The entire open reading frame (ORF) of the TYR is 1593 bp and encodes for 530 amino acids, which is well conserved compared with TYR of various species with higher degree of sequence similarity with other mammalian (74–99 %) than amphibian, aves, and fishes (56–73 %). The deduced amino acids contained one signal peptide, one transmembrane domain, five N-linked glycosylation sites, and two copper binding sites. The result of real-time quantitative PCR showed that the expression level of TYR was the highest in the dark-gray goats and the lowest in the light-gray ones, while the goats of dark-gray individuals have more than 50 % black fiber and light-gray ones less than 30 %. During the whole life of Jining gray goat, TYR expression level changes with certain regularity and their coat color will change correspondingly by investigating the expression level in ten development stages. After comparing the result and the coat phenotype, we presume that it seems to have a positive relationship between the color depth of coat and the expression level of TYR. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1275-1 Authors Weiyun Chen, College of Animal Sciences and Technology, Shandong Agricultural University, Taian, 271018 Shandong, People’s Republic of China Hui Wang, College of Animal Sciences and Technology, Shandong Agricultural University, Taian, 271018 Shandong, People’s Republic of China Bin Dong, College of Animal Sciences and Technology, Shandong Agricultural University, Taian, 271018 Shandong, People’s Republic of China Zhongdian Dong, College of Animal Sciences and Technology, Shandong Agricultural University, Taian, 271018 Shandong, People’s Republic of China Fenna Zhou, College of Animal Sciences and Technology, Shandong Agricultural University, Taian, 271018 Shandong, People’s Republic of China Yong Fu, College of Animal Sciences and Technology, Shandong Agricultural University, Taian, 271018 Shandong, People’s Republic of China Yongqing Zeng, College of Animal Sciences and Technology, Shandong Agricultural University, Taian, 271018 Shandong, People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Pleiotropic effects of serotonin (5-HT) in the cardiovascular system are well documented. However, it remains to be elucidated, whether 5-HT is present in adult mammalian cardiomyocytes. To address this issue, we investigated the levels of 5-HT in blood, plasma, platelets, cardiac tissue, and cardiomyocytes from adult mice and for comparison in human right atrial tissue. Immunohistochemically, 5-HT was hardly found in mouse cardiac tissue, but small amounts could be detected in renal preparations, whereas adrenal preparations revealed a strong positive immunoreaction for 5-HT. Using a sensitive HPLC detection system, 5-HT was also detectable in the mouse heart and human atrium. Furthermore, we could identify 5-HT in isolated cardiomyocytes from adult mice. These findings were supported by detection of the activity of 5-HT-forming enzymes—tryptophan hydroxylase and aromatic l -amino acid decarboxylase—in isolated cardiomyocytes from adult mice and by inhibition of these enzymes with p -chlorophenylalanine and 3-hydroxybenzyl hydrazine. Addition of the first intermediate of 5-HT generation, that is 5-hydroxytryptophan, enhanced the 5-HT level and inhibition of monoamine oxidase by tranylcypromine further increased the level of 5-HT. Our findings reveal the presence and synthesis of 5-HT in cardiomyocytes of the mammalian heart implying that 5-HT may play an autocrine and/or paracrine role in the heart. Content Type Journal Article Pages 1-12 DOI 10.1007/s11010-012-1270-6 Authors Klaus Pönicke, Medical Faculty, Institute of Pharmacology and Toxicology, Martin-Luther-University Halle-Wittenberg, Magdeburger Str. 4, 06112 Halle (Saale), Germany Ulrich Gergs, Medical Faculty, Institute of Pharmacology and Toxicology, Martin-Luther-University Halle-Wittenberg, Magdeburger Str. 4, 06112 Halle (Saale), Germany Igor B. Buchwalow, Institut für Hämatopathologie, Fangdieckstraße 75a, 22547 Hamburg, Germany Steffen Hauptmann, Medical Faculty, Institute of Pathology, Martin-Luther-University Halle-Wittenberg, 06097 Halle (Saale), Germany Joachim Neumann, Medical Faculty, Institute of Pharmacology and Toxicology, Martin-Luther-University Halle-Wittenberg, Magdeburger Str. 4, 06112 Halle (Saale), Germany Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    An investigation was made to reveal the protective effects of veratric acid (VA), a phenolic acid against atherogenic diet-induced hyperlipidemic rats. Male albino Wistar rats were fed with atherogenic diet (4% cholesterol, 1% cholic acid, and 0.5% 2-thiouracil) daily for 30 days and treated with VA (40 mg/kg body weight) daily for a period of 30 days. Rats fed with atherogenic diet showed significant ( P  〈 0.05) elevation in the level of plasma lipids, systolic and diastolic blood pressure, oxidative stress markers (thiobarbituric acid reactive substances, lipid peroxides) and significant ( P  〈 0.05) reduction in the activities of enzymatic (superoxide dismutase, catalase, glutathione peroxidase) and non-enzymatic (vitamin C, vitamin E, and reduced glutathione) antioxidants in erythrocytes, plasma, and tissues (liver, kidney, and aorta). Oral administration of VA (40 mg/kg body weight) for 30 days to atherogenic diet fed rats markedly attenuates systolic, diastolic blood pressure and lipid peroxidation products. Further, VA treatment significantly improved enzymatic and non-enzymatic antioxidants levels and showed beneficial effects on lipid profile in atherogenic diet rats. All the above alterations were supported by histopathological observations. These results indicate that oral administration of VA ameliorates atherogenic diet-induced hyperlipidemia in rats by its free radical scavenging; improving the antioxidants and lipid lowering properties. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1278-y Authors Boobalan Raja, Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalai Nagar, 608 002 Tamil Nadu, India Murugesan Saravanakumar, Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalai Nagar, 608 002 Tamil Nadu, India Gopal Sathya, Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalai Nagar, 608 002 Tamil Nadu, India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Reperfusion results in a rapid reintroduction of oxygen, glucose, and other restricted components to an ischemic tissue. It brings with it not only the necessary components for cell survival but also a burst of oxidative stress and cellular damage. In this study, our primary aims were to investigate glucose as a determining factor for the activation of the transcription factor NF-E2-related factor 2 (Nrf2) upon reperfusion and the expression of downstream anti-oxidant NADPH-dependent reductases. Exposure of renal epithelial HK-2 cells to oxygen and glucose reintroduction after depletion resulted in an increase in nuclear translocation of Nrf2 protein in a manner dependent upon glucose. This activation and the induction of the Nrf2-dependent gene NAD(P)H dehydrogenase, quinone 1 (NQO1) was observed to be maximum at a concentration of 5 mM glucose. Microarray analysis of mRNA from siRNA targeted cells under these conditions revealed the Nrf2-dependent expression of NADPH-dependent reductase enzymes NQO1, Aldo–keto reductase family 1, members C1-3 and dehydrogenase/reductase (SDR family) member 2 (DHRS2), all genes demonstrated to protect against oxidative stress-mediated cellular injury. In addition, NQO1 and DHRS2 mRNA levels were specifically upregulated on glucose reintroduction and were also increased in an in vivo ischemia reperfusion injury model of murine renal pedicle clamping. In conclusion, we demonstrate that glucose reintroduction after depletion activates Nrf2 and Nrf2 regulated NADPH-dependent reductase expression. We suggest these findings represent a previously unreported mechanism for the activation of Nrf2 as a cytoprotective pathway in IRI. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1300-4 Authors Daniel Crean, Conway Institute of Biomolecular and Biomedical Research, School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4, Ireland Luca Felice, Conway Institute of Biomolecular and Biomedical Research, School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4, Ireland Cormac T. Taylor, Conway Institute of Biomolecular and Biomedical Research, School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4, Ireland Hamid Rabb, Nephrology Division, Johns Hopkins University Hospital, Baltimore, MD, USA Paul Jennings, Division of Physiology, Department of Physiology and Medical Physics, Innsbruck Medical University, Innsbruck, Austria Martin O. Leonard, Conway Institute of Biomolecular and Biomedical Research, School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4, Ireland Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Adrenomedullin (ADM) and hypoxia-inducible factor-1α (HIF-1α) are important pro-proliferation genes in response to hypoxic stress. Although it was reported that ADM is a target gene for HIF-1, recent studies also showed that ADM regulates HIF-1 expression and its activity; however, the mechanism of action remains unknown. Two stable human endothelial cell lines with HIF-1α knockdown by hy926–siHIF-1α or HMEC–siHIF-1α were established. mRNA and protein expression of ADM and HIF-1α in EA.hy926 and HMEC1 cells were examined under hypoxic stress. Upon ADM treatment, cell proliferation was investigated and the expression profiles of HIF-1α and its target genes (VEGF, PFKP, PGK1, and AK1) were examined. Furthermore, the proline hydroxylase (PHD) mRNA level and its activity were investigated. We observed that mRNA and protein expression of ADM in hypoxia are earlier events than HIF-1α in EA.hy926 and HMEC1 cells. ADM-promoted cell proliferation of endothelial cells, which was HIF-1α dependent. We also found that ADM up-regulated the mRNA and protein expressions of HIF-1α- and HIF-1-targeted genes, and ADM up-regulated the protein expressions of HIF-1α through down-regulation of PHD mRNA expression and PHD activity. Content Type Journal Article Pages 1-11 DOI 10.1007/s11010-012-1267-1 Authors Li Chen, Burn Research Institute, Southwest Hospital, Third Military Medical University, Chongqing, 400038 China Ju-Hui Qiu, Burn Research Institute, Southwest Hospital, Third Military Medical University, Chongqing, 400038 China Ling-Ling Zhang, Burn Research Institute, Southwest Hospital, Third Military Medical University, Chongqing, 400038 China Xiang-Dong Luo, Burn Research Institute, Southwest Hospital, Third Military Medical University, Chongqing, 400038 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Roles of cyclooxygenase (COX) enzyme and intrinsic pathway of apoptosis have been explored for the chemopreventive effects of non-steroidal anti-inflammatory drugs (NSAIDs) on 9,10-dimethyl benz(a)anthracene (DMBA)-induced lung cancer in rat model. 16 weeks after the administration of DMBA, morphological analysis revealed the occurrences of tumours and lesions, which were regressed considerably with the co-administration of indomethacin and etoricoxib, the two NSAIDs under investigation. DMBA group was marked by hyperplasia and dysplasia as observed by histological examination, and these features were corrected to a large extent by the two NSAIDs. Elevated levels of COX-2 were seen in the DMBA group, the enzyme responsible for prostaglandin synthesis during inflammation and cancer, whilst the expression of the constitutive isoform, COX-1, was equally expressed in all the groups. Apoptosis was quantified by studying the activities of apaf-1, caspase-9, and 3 by immunofluorescence and western blots. Their activities were found to diminish in the DMBA-treated animals as compared to the other groups. Fluorescent co-staining of the isolated broncho-alveolar lavage cells showed reduced number of apoptotic cells in the DMBA group, indicating decrease in apoptosis after carcinogen administration. The present results thus suggest that the mechanism of cancer chemoprevention of NSAIDs may include the suppression of COX-2 and the induction of apoptosis. Content Type Journal Article Pages 1-11 DOI 10.1007/s11010-012-1286-y Authors Shruti Setia, Department of Biophysics, Panjab University, Chandigarh, 160014 India Vivek Vaish, Department of Biophysics, Panjab University, Chandigarh, 160014 India Sankar Nath Sanyal, Department of Biophysics, Panjab University, Chandigarh, 160014 India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    ER-to-Golgi protein transport involves transport vesicles of which formation is initiated by assembly of Sar1. The assembly of Sar1 is suppressed by protein kinase inhibitor H89, suggesting that ER-to-Golgi transport is regulated progressively by H89 sensitive kinase. ER-resident G i2 protein suppresses vesicle formation with inhibition of Sar1 assembly. This study examined whether these promotion and suppression of vesicle transport share the same signal pathway, by examining the effects of G i/o protein activator mastoparan 7 (Mp-7) and H89 on Sar1 and Sec23 recruitment onto microsomes. In a cell-free system for Sar1 translocation assay, GTPγS addition induced the translocation of Sar1 onto microsomes. Mp-7 and H89 decreased the Sar1 translocation. Double treatment of Mp-7 and H89 strongly decreased Sar1 translocation. In single and double treatments, however, G i/o protein inactivator pertussis toxin (IAP) partially restored the suppressive effect of Mp-7, but had not any effect on H89-induced effect. Then, the assembly of Sec23 onto the microsome was also increased by the addition of GTPγS. Sec23 translocation was decreased by Mp-7 and/or H89 treatment and recovered by IAP pretreatment except for H89 single treatment, similarly to Sar1 translocation in each treatment. Inhibitory effects of H89 and Mp-7on ER-to-Golgi vesicle transport by H89 or Mp-7 were also confirmed in a cell culture system by BFA-dispersion and BFA-reconstruction experiments. These findings indicate that promotion and suppression of ER-to-Golgi vesicle transport are modulated through separate signal pathways. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1295-x Authors Hiroshi Nakagawa, Laboratory of Toxicology, Course of Veterinary Science, Graduate School of Life and Environmental Biosciences, Osaka Prefecture University, 1-58, Rinku-Ourai-Kita, Izumisano, 598-8531 Japan Masakazu Ishizaki, Laboratory of Toxicology, Course of Veterinary Science, Graduate School of Life and Environmental Biosciences, Osaka Prefecture University, 1-58, Rinku-Ourai-Kita, Izumisano, 598-8531 Japan Shuichi Miyazaki, The Center for Advanced Research, Graduate School of Medical Sciences, Toho University of Medicine, Ota-ku, Omori-nishi 5-21-16, Tokyo, 143-8540 Japan Takuto Abe, Laboratory of Toxicology, Course of Veterinary Science, Graduate School of Life and Environmental Biosciences, Osaka Prefecture University, 1-58, Rinku-Ourai-Kita, Izumisano, 598-8531 Japan Kazuhiko Nishimura, Laboratory of Toxicology, Course of Veterinary Science, Graduate School of Life and Environmental Biosciences, Osaka Prefecture University, 1-58, Rinku-Ourai-Kita, Izumisano, 598-8531 Japan Masayuki Komori, Laboratory of Cellular and Molecular Biology, Course of Veterinary Science, Graduate School of Life and Environmental Biosciences, Osaka Prefecture University, 1-58, Rinku-Ourai-Kita, Izumisano, 598-8531 Japan Saburo Matsuo, Laboratory of Toxicology, Course of Veterinary Science, Graduate School of Life and Environmental Biosciences, Osaka Prefecture University, 1-58, Rinku-Ourai-Kita, Izumisano, 598-8531 Japan Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Because of their plasticity and availability, bone-marrow-derived mesenchymal stem cells (MSC) are a potential cell source for treating ischemic heart disease. Schwann cells (SC) play a critical role in neural remodeling and angiogenesis because of their secretion of cytokines such as vascular endothelial growth factor (VEGF). Cell microencapsulation, surrounding cells with a semipermeable polymeric membrane, is a promising tool to shelter cells from the recipient’s immune system. We investigated whether transplantation of microencapsulated SC (MC-SC) and MSC together could improve heart function by augmenting angiogenesis in acute myocardial infarction (AMI). Sprague–Dawley rats with ligation of the left anterior descending artery to induce AMI were randomly divided for cell transplantation into four groups—MC-SC+MSC, MC+MSC, MSC, MC-SC, and controls. Echocardiography was performed at 3 days and 2 and 4 weeks after AMI. Rat hearts were harvested on day 28 after transplantation and examined by immunohistochemistry and western blot analysis. Echocardiography revealed differences among the groups in fractional shortening and end-systolic and end-diastolic dimensions ( P  〈 0.05). The number of BrdU-positive cells was greater with MC-SC+MSC transplantation than the other groups ( P  〈 0.01). The vessel density and VEGF level in the infarcted zone was significantly increased with MC-SC+MSC transplantation ( P  〈 0.05). These results show that transplanting a combination of MC-SC and MSC could augment angiogenesis and improve heart function in AMI. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1291-1 Authors Yan Wang, Department of Cardiac Surgery, Provincial Hospital Affiliated to Shandong University, Shandong University, Jingwu Rd. 324, Jinan, 250021 People’s Republic of China Gang Zhang, Department of Cardiac Surgery, Provincial Hospital Affiliated to Shandong University, Shandong University, Jingwu Rd. 324, Jinan, 250021 People’s Republic of China Yongbo Hou, Department of Cardiac Surgery, Provincial Hospital Affiliated to Shandong University, Shandong University, Jingwu Rd. 324, Jinan, 250021 People’s Republic of China Jian Chen, Key Laboratory of Smart Drug Delivery, Ministry of Education & PLA School of Pharmacy, Fudan University, Shanghai, 201203 People’s Republic of China Juan Wang, Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Public Health, Shandong University Qilu Hospital, Jinan, 250012 Shandong, People’s Republic of China Chengwei Zou, Department of Cardiac Surgery, Provincial Hospital Affiliated to Shandong University, Shandong University, Jingwu Rd. 324, Jinan, 250021 People’s Republic of China Decai Li, Department of Cardiac Surgery, Provincial Hospital Affiliated to Shandong University, Shandong University, Jingwu Rd. 324, Jinan, 250021 People’s Republic of China Hongxin Li, Department of Cardiac Surgery, Provincial Hospital Affiliated to Shandong University, Shandong University, Jingwu Rd. 324, Jinan, 250021 People’s Republic of China Qian Zhang, Department of Cardiac Surgery, Provincial Hospital Affiliated to Shandong University, Shandong University, Jingwu Rd. 324, Jinan, 250021 People’s Republic of China Anbiao Wang, Department of Cardiac Surgery, Provincial Hospital Affiliated to Shandong University, Shandong University, Jingwu Rd. 324, Jinan, 250021 People’s Republic of China Quanxin Fan, Department of Cardiac Surgery, Provincial Hospital Affiliated to Shandong University, Shandong University, Jingwu Rd. 324, Jinan, 250021 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    There is growing evidence suggesting that circulating fibrocytes (CFs) play a pivotal role in tissue repair and fibrosis. In contrast, in recent studies, angiotensin-(1-7) [Ang-(1-7)] has been shown to antagonize fibrosis. The purpose of this study was to examine the direct effect of Ang-(1-7) on CFs. Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation. Using laser scanning confocal microscopy, CFs were identified as adherent cells that stained positive for both CD34 and collagen-I. After 14 days of culture, CFs were stimulated with Ang-(1-7) at concentrations of 10 nM, 100 nM, 1 μM or 10 μM, in the absence and presence of pretreatment with A-779, N G -nitro- l -arginine methyl ester (L-NAME) or both, for 24, 48 or 72 h. The number of cells, cellular proliferation, and level of apoptosis were determined by hematoxylin and eosin staining, the Cell Counting Kit-8 (CCK8) assay and the annexin V/propidium iodide binding assay, respectively. The collagen content of CFs was measured by the concentration of hydroxyproline, which was detected using the enzymatic digestion method. The expression of endothelial nitric oxide synthase (eNOS) was assayed by western Blot analysis, while nitric oxide (NO) generation was detected using the Griess method. We found that Ang-(1-7) increases apoptosis and eNOS/NO production in CFs. In addition, Ang-(1-7) decreases the number, proliferative capacity and collagen-secretion of CFs in a concentration- and time-dependent manner. These data suggest that Ang-(1-7) suppresses the both the number and function of CFs possibly by increasing eNOS/NO production in the CFs. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1223-0 Authors Kan Wang, Institute of Cardiology, The First Affiliated Hospital, School of Medicine, Zhejiang University, No. 79, Qingchun Road, Hangzhou, 310003 People’s Republic of China Xiaosheng Hu, Institute of Cardiology, The First Affiliated Hospital, School of Medicine, Zhejiang University, No. 79, Qingchun Road, Hangzhou, 310003 People’s Republic of China Changqing Du, Institute of Cardiology, The First Affiliated Hospital, School of Medicine, Zhejiang University, No. 79, Qingchun Road, Hangzhou, 310003 People’s Republic of China Shike Tu, Institute of Cardiology, The First Affiliated Hospital, School of Medicine, Zhejiang University, No. 79, Qingchun Road, Hangzhou, 310003 People’s Republic of China Furong Zhang, Institute of Cardiology, The First Affiliated Hospital, School of Medicine, Zhejiang University, No. 79, Qingchun Road, Hangzhou, 310003 People’s Republic of China Xudong Xie, Institute of Cardiology, The First Affiliated Hospital, School of Medicine, Zhejiang University, No. 79, Qingchun Road, Hangzhou, 310003 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Treatment during early tumor development has greater success because tissue growth remains largely confined to its original locus. At later stages, malignant cells migrate from their original location, invade surrounding normal areas, and can disseminate widely throughout the body. Remodeling of the extracellular matrix (ECM) serves as a key facilitator of this dissemination. Proteolytic enzymes including plasmin and matrix metalloproteinases (MMPs) play an integral role in degrading the surrounding ECM proteins and clearing a path for tumor cell migration. Specific MMPs are highly expressed late during malignant tumor invasion. It is not understood whether early changes in MMPs influence apoptotic and necrotic cell death, processes known to govern the early stages of carcinogenesis. Similarly, the interaction between MDM2 and p53 is tightly controlled by a complex array of post-translational modifications, which in turn dictates the stability and activity of both p53 and MDM2. The present studies examine the hypothesis that model hepatotoxin dimethylnitrosamine (DMN), which is also a model carcinogen, will induce the MMP family of proteins after administration in hepatotoxic doses. Doses of 25, 50, and 100 mg/kg DMN were administered i.p. to male C3H mice. Changes in parameters associated with apoptotic and necrotic cell death, DNA damage, cell proliferation, and extracellular proteinases were examined in liver at 24 h. Serum ALT activity, oxidative stress [malondialdehyde], and caspase-activated DNAse mediated DNA laddering increased in a dose-dependent manner, as did the level of MDM2 protein. MMP-9, -10 and -12 (gelatinase-B, stromelysin-2, macrophage elastase), and p53 protein levels increased following 25 mg/kg DMN, but were successively decreased after higher DMN doses. The results of this study demonstrate changes in MDM2 and MMPs during DMN-induced acute liver injury and provide a plausible linkage between DMN-induced oxidative stress-mediated genomic injury and its likely involvement in setting the stage for initiating subsequent metastatic disease at later circumstances. Content Type Journal Article Pages 1-11 DOI 10.1007/s11010-012-1277-z Authors Ismail Syed, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Ave, CLS-0747, Boston, MA 02215, USA Jasmine Rathod, Division of Pharmaceutical Sciences, Manchester College school of Pharmacy, Fort Wayne, IN 46805, USA Mayur Parmar, Division of Pharmaceutical Sciences, Manchester College school of Pharmacy, Fort Wayne, IN 46805, USA George B. Corcoran, Department of Pharmaceutical Sciences, Eugene Applebaum College of Pharmacy and Health Sciences, Wayne State University, Detroit, MY 48201-2417, USA Sidhartha D. Ray, Division of Pharmaceutical Sciences, Manchester College school of Pharmacy, Fort Wayne, IN 46805, USA Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Central to the maintenance of genomic integrity is the cellular DNA damage response. Depending on the type of genotoxic stress and through the activation of multiple signaling cascades, it can lead to cell cycle arrest, DNA repair, senescence, and apoptosis. p19INK4d, a member of the INK4 family of CDK inhibitors, plays a dual role in the DNA damage response, inhibiting cell proliferation and promoting DNA repair. Consistently, p19INK4d has been reported to become upregulated in response to UV irradiation and a great variety of genotoxic agents. Here, this induction is shown to result from a transcriptional stimulatory mechanism that can occur at every phase of the cell cycle except during mitosis. Moreover, evidence is presented that demonstrates that E2F1 is involved in the induction of p19INK4d following UV treatment, as it is prevented by E2F1 protein ablation and DNA-binding inhibition. Specific inhibition of this regulation using triplex-forming oligonucleotides that target the E2F response elements present in the p19INK4d promoter also block p19INK4d upregulation and sensitize cells to DNA damage. These results constitute the first description of a mechanism for the induction of p19INK4d in response to UV irradiation and demonstrate the physiological relevance of this regulation following DNA damage. Content Type Journal Article Pages 1-7 DOI 10.1007/s11010-012-1289-8 Authors Abel L. Carcagno, Laboratorio de Biología Molecular, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria Pabellón II Piso 4, 1428 Ciudad de Buenos Aires, Argentina Luciana E. Giono, Laboratorio de Biología Molecular, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria Pabellón II Piso 4, 1428 Ciudad de Buenos Aires, Argentina Mariela C. Marazita, Laboratorio de Biología Molecular, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria Pabellón II Piso 4, 1428 Ciudad de Buenos Aires, Argentina Daniela S. Castillo, Laboratorio de Biología Molecular, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria Pabellón II Piso 4, 1428 Ciudad de Buenos Aires, Argentina Nicolás Pregi, Laboratorio de Biología Molecular, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria Pabellón II Piso 4, 1428 Ciudad de Buenos Aires, Argentina Eduardo T. Cánepa, Laboratorio de Biología Molecular, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria Pabellón II Piso 4, 1428 Ciudad de Buenos Aires, Argentina Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Methionine aminopeptidase (MAP) performs the essential post-translational N-terminal methionine excision (NME) of nascent polypeptides during protein synthesis. To characterize MAP from Mycobacterium tuberculosis , two homolgues, map A (Rv0734) and map B (Rv2861c), were over expressed and purified as recombinant proteins in E.   coli . In vitro activity assay of apo-MtbMAPs using l -Met- p -nitro anilide as substrate revealed MtbMAP A to be catalytically more efficient compared to MtbMAP B. Ni 2+ was the best activator of apo-MtbMAP A, whereas Ni 2+ and Co 2+ activated apo-MtbMAP B equally. MtbMAP B showed higher thermo-stability, but was feedback inhibited by higher concentrations of l -methionine. Aminopeptidase inhibitors like actinonin and bestatin inhibited both MtbMAPs, more prominently MtbMAP B. Among the site-directed mutants of MtbMAP B, substitution of metal-binding residue D142 completely abolished enzyme activity, whereas substitution of residues forming S1′ pocket, C105S and T94C, had only moderate effects on substrate hydrolysis. Present study identified a specific insertion region in MtbMAP A sequence which differentiates it from other bacterial and eukaryotic MAPs. A deletion mutant lacking amino acids from this insertion region (MtbMAP A-∆164-176) was constructed to probe into their structural and functional role in activity and stability of MtbMAP A. The limited success in soluble expression of this deletion mutant suggests further optimizations of expression conditions or alternative bioinformatics approaches for further characterization of this deletion mutant of MtbMAP A. Content Type Journal Article Pages 1-12 DOI 10.1007/s11010-012-1260-8 Authors Sai Shyam Narayanan, Biotechnology Division, National Institute for Interdisciplinary Science and Technology (NIIST), CSIR, Trivandrum, 695019 Kerala, India Kesavan Madhavan Nampoothiri, Biotechnology Division, National Institute for Interdisciplinary Science and Technology (NIIST), CSIR, Trivandrum, 695019 Kerala, India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Cyclic ADP-ribose (cADPR), a metabolite of NAD + , is known to function as a second messenger for intracellular Ca 2+ mobilization in various vertebrate and invertebrate tissues. In this study, we isolated two Xenopus laevis cDNAs (frog cd38 and cd157 cDNAs) homologous to the one encoding the human cADPR-metabolizing enzyme CD38. Frog CD38 and CD157 are 298-amino acid proteins with 35.9 and 27.2 % identity to human CD38 and CD157, respectively. Transfection of expression vectors for frog CD38 and CD157 into COS-7 cells revealed that frog CD38 had NAD + glycohydrolase, ADP-ribosyl cyclase (ARC), and cADPR hydrolase activities, and that frog CD157 had no enzymatic activity under physiological conditions. In addition, when recombinant CD38 and frog brain homogenate were electrophoresed on an SDS–polyacrylamide gel, ARC of the brain homogenate migrated to the same position in the gel as that of frog CD38, suggesting that frog CD38 is the major enzyme responsible for cADPR metabolism in amphibian cells. The frog cd38 gene consists of eight exons and is ubiquitously expressed in various tissues. These findings provide evidence for the existence of the CD38–cADPR signaling system in frog cells and suggest that the CD38–cADPR signaling system is conserved during vertebrate evolution. Content Type Journal Article Pages 1-12 DOI 10.1007/s11010-012-1284-0 Authors Takayuki Ikeda, Department of Biochemistry, Kanazawa Medical University School of Medicine, 1-1 Daigaku, Uchinada, Kahoku-gun, Ishikawa, 920-0293 Japan Shin Takasawa, Department of Biochemistry, Nara Medical University, Kashihara, Nara, 634-8521 Japan Naoya Noguchi, Department of Advanced Biological Sciences for Regeneration (Kotobiken Medical Laboratories), Tohoku University Graduate School of Medicine, Sendai, Miyagi, 980-8575 Japan Koji Nata, Department of Medical Biochemistry, Iwate Medical University School of Pharmacy, 2-1-1 Nishitokuda, Yahaba-cho, Shiwa-gun, Iwate, 028-3603 Japan Akiyo Yamauchi, Department of Biochemistry, Nara Medical University, Kashihara, Nara, 634-8521 Japan Iwao Takahashi, Department of Medical Biochemistry, Iwate Medical University School of Pharmacy, 2-1-1 Nishitokuda, Yahaba-cho, Shiwa-gun, Iwate, 028-3603 Japan Takeo Yoshikawa, Department of Advanced Biological Sciences for Regeneration (Kotobiken Medical Laboratories), Tohoku University Graduate School of Medicine, Sendai, Miyagi, 980-8575 Japan Akira Sugawara, Department of Advanced Biological Sciences for Regeneration (Kotobiken Medical Laboratories), Tohoku University Graduate School of Medicine, Sendai, Miyagi, 980-8575 Japan Hideto Yonekura, Department of Biochemistry, Kanazawa Medical University School of Medicine, 1-1 Daigaku, Uchinada, Kahoku-gun, Ishikawa, 920-0293 Japan Hiroshi Okamoto, Tohoku University, 2-1-1 Katahira, Sendai, Miyagi, 980-8577 Japan Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    To evaluate the role of SNARE proteins in the constitutive exocytosis, we knocked down syntaxin 3, 4, 5, 6, 7, and VAMP3, 5, 7, 8 with their siRNAs, and determined the cell-to-medium ratio of CLuc, a secreted luciferase of Cypridina noctiluca . Although the protein level of SNAREs in HeLa cells was markedly reduced by the siRNA treatment, the cell/medium ratio was scarcely increased by any siRNAs except for syntaxin 5. The accumulation of GFP-tagged human growth hormone was also visible only by the knockdown of syntaxin 5. To examine whether the residual amount of SNAREs are sufficient for maintaining normal constitutive exocytosis, we estimated the effect of siRNAs on the level of post-Golgi SNARE complexes containing syntaxin 4, SNAP23, and VAMP3 or VAMP8. The amount of SNARE complexes was robustly decreased by siRNAs and was well correlated with the residual amount of SNAREs in the lysates, suggesting that SNAREs are unnecessarily excessive for the formation of post-Golgi SNARE complexes in HeLa cells. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1293-z Authors Miki Okayama, Department of Orthodontics, School of Dentistry, Health Sciences University of Hokkaido, Tobetsu, Hokkaido 061-0293, Japan Akiko Shitara, Department of Biochemistry, School of Dentistry, Health Sciences University of Hokkaido, Tobetsu, Hokkaido 061-0293, Japan Toshiya Arakawa, Department of Biochemistry, School of Dentistry, Health Sciences University of Hokkaido, Tobetsu, Hokkaido 061-0293, Japan Yoshifumi Tajima, Department of Oral Pathology, School of Dentistry, Meikai University, Sakado, Saitama 350-0283, Japan Itaru Mizoguchi, Department of Orthodontics, School of Dentistry, Health Sciences University of Hokkaido, Tobetsu, Hokkaido 061-0293, Japan Taishin Takuma, Department of Biochemistry, School of Dentistry, Health Sciences University of Hokkaido, Tobetsu, Hokkaido 061-0293, Japan Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Previous studies have shown that Wnt signaling is involved in postnatal mammalian myogenesis; however, the downstream mechanism of Wnt signaling is not fully understood. This study reports that the murine four-and-a-half LIM domain 1 ( Fhl1 ) could be stimulated by β-catenin or LiCl treatment to induce myogenesis. In contrast, knockdown of the Fhl1 gene expression in C2C12 cells led to reduced myotube formation. We also adopted reporter assays to demonstrate that either β-catenin or LiCl significantly activated the Fhl1 promoter, which contains four putative consensus TCF/LEF binding sites. Mutations of two of these sites caused a significant decrease in promoter activity by luciferase reporter assay. Thus, we suggest that Wnt signaling induces muscle cell differentiation, at least partly, through Fhl1 activation. Content Type Journal Article Pages 1-12 DOI 10.1007/s11010-012-1266-2 Authors Jing-Yu Lee, Department of Bioagricultural Sciences, National Chiayi University, No. 300 Syuefu Rd., Chiayi, 60004 Taiwan, ROC I-Chun Chien, Department of Bioagricultural Sciences, National Chiayi University, No. 300 Syuefu Rd., Chiayi, 60004 Taiwan, ROC Win-Yu Lin, Department of Bioagricultural Sciences, National Chiayi University, No. 300 Syuefu Rd., Chiayi, 60004 Taiwan, ROC Shao-min Wu, Department of Bioagricultural Sciences, National Chiayi University, No. 300 Syuefu Rd., Chiayi, 60004 Taiwan, ROC Bo-Huei Wei, Department of Bioagricultural Sciences, National Chiayi University, No. 300 Syuefu Rd., Chiayi, 60004 Taiwan, ROC Yu-En Lee, Department of Bioagricultural Sciences, National Chiayi University, No. 300 Syuefu Rd., Chiayi, 60004 Taiwan, ROC Hu-Hui Lee, Department of Bioagricultural Sciences, National Chiayi University, No. 300 Syuefu Rd., Chiayi, 60004 Taiwan, ROC Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    β-1,4-galactosyltransferase-I (β-1,4-GalT-I) plays a critical role in the initiation and maintenance of peripheral nervous system inflammatory reaction. However, the exact function of β-1,4-GalT-I in the regulation of SCs proliferation and apoptosis remains unclear. In this study, we found that low concentration of tumor necrosis factor-alpha (TNF-α) induced SCs proliferation, while high concentration of TNF-α induced SCs apoptosis. Meanwhile, the expressions of β-1,4-GalT-I, TNFR1, and TNFR2 were changed following. When β-1,4-GalT I overexpression, low concentration of TNF-α-induced SCs proliferation was partially repressed. Concurrently, the activity of ERK1/2 was decreased. While knocking down β-1,4-GalT I expression, high concentration of TNF-α-induced SCs apoptosis was partially rescued. Consistent with this, the activity of P38 and JNK were decreased. We also found anti-TNFR2 antibody suppressed low concentration of TNF-α-induced SCs proliferation, while anti-TNFR1 antibody inhibited high concentration of TNF-α-induced SCs apoptosis. Thus, present data show that β-1,4-GalT I may play an important role in SCs proliferation and apoptosis induced by TNF-α via different signal pathways and TNFR. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1254-6 Authors Qin Yuan, Department of Immunology, Medical College, Nantong University, Nantong, Jiangsu 226001, People’s Republic of China Huiguang Yang, Department of Osteology, Affiliated Jiangyin Hospital of Nantong University, Nantong, Jiangsu 226001, People’s Republic of China Chun Cheng, Department of Immunology, Medical College, Nantong University, Nantong, Jiangsu 226001, People’s Republic of China Chunmiao Li, Department of Immunology, Medical College, Nantong University, Nantong, Jiangsu 226001, People’s Republic of China Xiujie Wu, Department of Immunology, Medical College, Nantong University, Nantong, Jiangsu 226001, People’s Republic of China Weipeng Huan, Department of Immunology, Medical College, Nantong University, Nantong, Jiangsu 226001, People’s Republic of China Huiqing Sun, Department of Osteology, Affiliated Jiangyin Hospital of Nantong University, Nantong, Jiangsu 226001, People’s Republic of China Zhengming Zhou, Department of Osteology, Affiliated Jiangyin Hospital of Nantong University, Nantong, Jiangsu 226001, People’s Republic of China Yingying Wang, Department of Immunology, Medical College, Nantong University, Nantong, Jiangsu 226001, People’s Republic of China Yunhong Zhao, Department of Immunology, Medical College, Nantong University, Nantong, Jiangsu 226001, People’s Republic of China Xiang Lu, Department of Geriatrics, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210011, People’s Republic of China Aiguo Shen, The Jiangsu Province Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu 226001, People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Antiestrogen is one type of the endocrine therapeutic agents for estrogen receptor α (ERα)-positive breast cancer. Unfortunately, this treatment alone is insufficient. Here we reported a novel potential anticancer strategy by using histone deacetylase (HDAC) inhibitor to enhance the action of endocrine therapy in ERα-positive breast cancer cell. The well-described HDAC inhibitor, trichostatin A (TSA), and antiestrogen raloxifene were found to, respectively, inhibit E2-induced proliferation of MCF-7 breast cancer cell in a dose-responsive and time-dependent manner. TSA and raloxifene enhanced the antiproliferative activity of each other by promoting cell death via apoptosis and cell cycle arrest. Thus, they displayed better antiproliferative effects in combined treatment than that with either agent alone. The expression level of estrogen receptor β (ERβ) showed a marked increase after TSA or/and raloxifene treatment. Treatments with TSA or/and raloxifene resulting in the up-regulation of ERβ are in accordance with the antiproliferative effects of the two agents. Furthermore, the over-expression of ERβ by adenovirus delivery could inhibit the proliferation of MCF-7 tumor cells and drastically enhanced the antiproliferative effects of TSA and raloxifene. These results demonstrated that the interference of ERβ on the antiproliferative effects of HDAC inhibitor and antiestrogen constitutes a promising approach for breast cancer treatment. Content Type Journal Article Pages 1-12 DOI 10.1007/s11010-012-1288-9 Authors Zhenzhen Tu, Department of Biomedical Engineering, School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210009 People’s Republic of China Hui Li, Department of Biomedical Engineering, School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210009 People’s Republic of China Yuxiang Ma, Department of Biomedical Engineering, School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210009 People’s Republic of China Bin Tang, Department of Biomedical Engineering, School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210009 People’s Republic of China Junmei Tian, Department of Biomedical Engineering, School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210009 People’s Republic of China Walter Akers, Department of Radiology, School of Medicine, Washington University in St. Louis, St. Louis, MO 63110, USA Samuel Achilefu, Department of Radiology, School of Medicine, Washington University in St. Louis, St. Louis, MO 63110, USA Yueqing Gu, Department of Biomedical Engineering, School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210009 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Bone homeostasis is regulated through osteoclasts and osteoblasts. Osteoporosis, which is induced with its accompanying decrease in bone mass with increasing age, is widely recognized as a major public health problem. Bone loss may be due to decreased osteoblastic bone formation and increased osteoclastic bone resorption. There is growing evidence that nutritional and food factors may play a part in the prevention of bone loss with aging and have been to be worthy of notice in the prevention of osteoporosis. Zinc, an essential trace element, or genistein, which are contained in soybeans, has been shown to have a stimulatory effect on osteoblastic bone formation and an inhibitory effect on osteoclastic bone resorption, thereby increasing bone mass. These factors have an effect on protein synthesis and gene expression, which are related to bone formation in osteoblastic cells and bone resorption in osteoclastic cells. The combination of zinc and genistein is found to reveal the synergistic effect on bone anabolic effect. The oral administration of those factors has been shown to prevent on bone loss in ovariectomized rats, an animal model for osteoporosis, indicating a role in the prevention of osteoporosis. Supplemental intake of ingredient with the combination of zinc and genistein has been shown to have a preventive effect on osteoporosis in human subjects, suggesting a role in the prevention of bone loss. Content Type Journal Article Pages 1-21 DOI 10.1007/s11010-012-1298-7 Authors Masayoshi Yamaguchi, Department of Foods and Nutrition, The University of Georgia, 425 River Road, Rhodes Center, Room 448, Athens, GA 30602-2771, USA Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The aim of this study was to investigate the expression of ZEB1 in gastric carcinoma, its correlation with the clinicopathology of gastric carcinoma, and the role of ZEB1 in invasion and metastasis in gastric carcinoma. ZEB1 expression was analyzed by immunohistochemistry and Western blot in 45 gastric carcinoma tissue samples that contained the adjacent gastric mucosa. The correlation between ZEB1 expression, the occurrence and development of gastric cancer, and clinical pathology was investigated. ZEB1 expression in the human gastric carcinoma cell line AGS was downregulated by RNA interference, and changes in ZEB1 expression corresponded with changes in the invasive and metastatic ability of AGS cells. Immunohistochemistry revealed that ZEB1 protein expression in gastric carcinoma tissues was significantly higher than in normal gastric mucosa tissues ( p  〈 0.001). A lower degree of differentiation of gastric cancer ( p  = 0.009), a higher TNM (tumor, node, and metastasis) stage ( p  = 0.010), and a larger scope of invasion were correlated with higher expression of ZEB1 ( p  = 0.041, 0.002). However, the expression of ZEB1 in gastric carcinoma tissue was independent of gender, age, and tumor size ( p  〉 0.05). Western blot results also showed that ZEB1 protein expression was significantly higher in gastric carcinoma tissue than in the adjacent normal gastric mucosa tissue ( p  = 0.008). A lower degree of differentiation of the gastric carcinoma correlated with a higher TNM stage, and a larger scope of invasion correlated with increased ZEB1 expression ( p  = 0.023). Transfection of ZEB1 siRNA in AGS cells significantly decreased the expression level of ZEB1 protein ( p  = 0.035). Furthermore, the number of cells that could pass through the Transwell chamber was significantly lower in the transfected group than in the non-transfected control group ( p  = 0.039), indicating that the suppression of ZEB1 expression could significantly reduce the invasive and metastatic ability of AGS cells ( p  = 0.005). Concluding, in gastric carcinoma tissue, overexpression of ZEB1 may be related to the occurrence and development as well as invasion and metastasis of gastric carcinoma. Content Type Journal Article Pages 1-7 DOI 10.1007/s11010-012-1299-6 Authors Baoqing Jia, Department of Surgical Oncology, General Hospital of Chinese People’s Liberation Army, No. 28, Fuxing Rd, Beijing, 100853 People’s Republic of China Hongyi Liu, Department of Surgical Oncology, General Hospital of Chinese People’s Liberation Army, No. 28, Fuxing Rd, Beijing, 100853 People’s Republic of China Qinglong Kong, Department of Surgical Oncology, General Hospital of Chinese People’s Liberation Army, No. 28, Fuxing Rd, Beijing, 100853 People’s Republic of China Bing Li, Department of Surgical Oncology, General Hospital of Chinese People’s Liberation Army, No. 28, Fuxing Rd, Beijing, 100853 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    In general, oxidative stress resulting from an imbalance between prooxidant and antioxidant systems plays an important role in the pathogenesis of cancer. Morin (3,5,7,2′,4′-pentahydroxyflavone), a member of the flavanol group, has been shown to possess chemopreventive potential against hepatocellular and colon cancer in experimental animals. Given the demonstrated importance of morin, aim of the present study was to evaluate the effect of morin on antiproliferative and anticarcinogenic effect against DMBA-induced experimental mammary carcinogenesis. Oral administration of 7,12-dimethylbenz(a)-anthracene (25 mg/kg body weight) to rats resulted in significant reduction of body weight, enzymic antioxidants (superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase), and nonenzymic antioxidants (reduced glutathione, vitamin C, and vitamin E). The levels of lipid peroxidation markers (thiobarbituric acid reactive substances and hydroperoxides) and tumor markers such as CA 15-3, AFP and CEA in serum were increased significantly in cancer-induced animals as compared to control rats. Oral supplementation of morin at a dose of 50 mg/kg body weight significantly improved the body weight, enzymic, and nonenzymic antioxidants and considerably decreased the lipid peroxidation marker and tumor markers levels. Histological observations also correlated with the biochemical parameters. Tumor bearing animals showed marked increase in proliferating cell nuclear antigen-positive cells and also the number of AgNOR/nuclei compared with control rats while this expression levels were significantly reduced upon morin treatment. Thus, this study reveals the possible beneficial effect of morin as chemopreventive agent against the oxidative stress induced during mammary carcinogenesis. Content Type Journal Article Pages 79-92 DOI 10.1007/s11010-011-1207-5 Authors Ramadass Nandhakumar, Department of Biochemistry, University of Madras, Guindy Campus, Chennai, 600025 Tamil Nadu, India Kombiyil Salini, Department of Biochemistry, University of Madras, Guindy Campus, Chennai, 600025 Tamil Nadu, India Sivasithambaram Niranjali Devaraj, Department of Biochemistry, University of Madras, Guindy Campus, Chennai, 600025 Tamil Nadu, India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177 Journal Volume Volume 364 Journal Issue Volume 364, Numbers 1-2

Description:    In this study, we investigated the role of GABAergic and glutamatergic systems in the anticonvulsant action of 3-alkynyl selenophene (3-ASP) in a pilocarpine (PC) model of seizures. To this purpose, 21 day-old rats were administered with an anticonvulsant dose of 3-ASP (50 mg/kg, per oral, p.o.), and [ 3 H]γ-aminobutyric acid (GABA) and [ 3 H]glutamate uptakes were carried out in slices of cerebral cortex and hippocampus. [ 3 H]GABA uptake was decreased in cerebral cortex (64%) and hippocampus (58%) slices of 21 day-old rats treated with 3-ASP. In contrast, no alteration was observed in [ 3 H]glutamate uptake in cerebral cortex and hippocampus slices of 21 day-old rats that received 3-ASP. Considering the drugs that increase synaptic GABA levels, by inhibiting its uptake or catabolism, are effective anticonvulsants, we further investigated the possible interaction between sub-effective doses of 3-ASP and GABA uptake or GABA transaminase (GABA-T) inhibitors in PC-induced seizures in 21 day-old rats. For this end, sub-effective doses of 3-ASP (10 mg/kg, p.o.) and dl -2,4-diamino- n -butyric acid hydrochloride (DABA, an inhibitor of GABA uptake—2 mg/kg, intraperitoneally; i.p.) or aminooxyacetic acid hemihydrochloride (AOAA; a GABA-T inhibitor—10 mg/kg, i.p.) were co-administrated to 21 day-old rats before PC (400 mg/kg; i.p.) treatment, and the appearance of seizures was recorded. Results demonstrated that treatment with AOAA and 3-ASP or DABA and 3-ASP significantly abolished the number of convulsing animals induced by PC. The present study indicates that 3-ASP reduced [ 3 H]GABA uptake, suggesting that its anticonvulsant action is related to an increase in inhibitory tonus. Content Type Journal Article Pages 1-6 DOI 10.1007/s11010-012-1257-3 Authors Ethel Antunes Wilhelm, Laboratório de Síntese, Reatividade e Avaliação Farmacológica e Toxicológica de Organocalcogênios, Centro de Ciências Naturais e Exatas, Universidade Federal de Santa Maria, Santa Maria, RS 97105-900, Brazil Bibiana Mozzaquatro Gai, Laboratório de Síntese, Reatividade e Avaliação Farmacológica e Toxicológica de Organocalcogênios, Centro de Ciências Naturais e Exatas, Universidade Federal de Santa Maria, Santa Maria, RS 97105-900, Brazil Ana Cristina Guerra Souza, Laboratório de Síntese, Reatividade e Avaliação Farmacológica e Toxicológica de Organocalcogênios, Centro de Ciências Naturais e Exatas, Universidade Federal de Santa Maria, Santa Maria, RS 97105-900, Brazil Cristiani Folharini Bortolatto, Laboratório de Síntese, Reatividade e Avaliação Farmacológica e Toxicológica de Organocalcogênios, Centro de Ciências Naturais e Exatas, Universidade Federal de Santa Maria, Santa Maria, RS 97105-900, Brazil Juliano Alex Roehrs, Laboratório de Síntese, Reatividade e Avaliação Farmacológica e Toxicológica de Organocalcogênios, Centro de Ciências Naturais e Exatas, Universidade Federal de Santa Maria, Santa Maria, RS 97105-900, Brazil Cristina Wayne Nogueira, Laboratório de Síntese, Reatividade e Avaliação Farmacológica e Toxicológica de Organocalcogênios, Centro de Ciências Naturais e Exatas, Universidade Federal de Santa Maria, Santa Maria, RS 97105-900, Brazil Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    In hypertensive animals and patients, oxidative stress represents the primary risk factor for progression of left ventricular hypertrophy. Recently, it has been demonstrated that hydrogen, as a novel antioxidant, can selectively reduce hydroxyl radicals and peroxynitrite anion to exert therapeutic antioxidant activity. In the current study, we explored the effect of chronic treatment with hydrogen-rich saline (HRS) on left ventricular hypertrophy in spontaneously hypertensive rats (SHR). The 8-week-old male SHR and age-matched Wistar-Kyoto rats (WKY) were randomized into HRS-treated (6 ml/kg/day for 3 months, i.p.) and vehicle-treated groups. HRS treatment had no significant effect on blood pressure, but it effectively attenuated left ventricular hypertrophy in SHR. HRS treatment abated oxidative stress, restored the activity of antioxidant enzymes including GPx, GST, catalase, and SOD, suppressed NADPH oxidase activity and downregulated Nox2 and Nox4 expression in left ventricles of SHR. HRS treatment suppressed pro-inflammatory cytokines including IL-1β, IL-6, TNF-α, and MCP-1, and inhibited NF-κB activation through preventing IκBα degradation in left ventricles of SHR. HRS treatment preserved mitochondrial function through restoring electron transport chain enzyme activity, repressing ROS formation, and enhancing ATP production in left ventricles of SHR. Moreover, HRS treatment suppressed ACE expression and locally reduced angiotensin II generation in left ventricles of SHR. In conclusion, HRS treatment attenuates left ventricular hypertrophy through abating oxidative stress, suppressing inflammatory process, preserving mitochondrial function, in which suppression of HRS on angiotensin II in left ventricles locally might be involved. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1264-4 Authors Yong-Sheng Yu, Department of Pharmacology, College of Pharmacy, Second Military Medical University, Shanghai, 200433 China Hao Zheng, Department of Pharmaceutical Science and Technology, College of Chemistry and Biology, Donghua University, Shanghai, China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    In the earlier study, sodium orthovanadate (SOV) has been reported to be a powerful inhibitor of (Na + , K + ) adenosine triphosphatase, exhibit widespread actions on the renal and cardiovascular systems, induces smooth muscle contraction by inhibiting the phosphorylation of the protein tyrosine phosphatases. In the current study, we aimed to investigate the cellular mechanisms by which SOV facilitated contractile response of vas deferens smooth muscle and its potential therapeutic advantage. Exogenous application of ATP and NA-caused contraction was strengthened by pretreatment with SOV. This facilitation was inhibited not by bath with the inhibitor of P2 receptor, PPADS, or the inhibitor of α1 receptor, Prazosin, but by bath with the protein tyrosine kinase inhibitor, Genistein. SOV induced a sustained increase in intracellular Ca 2+ of smooth muscle cells, which was abolished by 100 μM Genistein or Ca 2+ -free solution. The facilitation of SOV could also be inhibited by the selective inhibitors of TRP channel, 2-APB and non-selective cation channel, Gd 3+ , Ni + . The in vivo study showed that peritoneal injection of SOV in dystrophic mice (mdx mice) enhanced the contraction of vas deferens smooth muscle stimulated by electrical field stimulation, ATP, noradrenaline, or KCl. The above results suggest that SOV facilitates the concentration of vas deferens smooth muscle through the tyrosine phosphorylation activated the non-selective cation channels, which has potential use in the therapy for muscle dysfunction. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1292-0 Authors Lei Zhao, Department of Physiology, Guangzhou Medical University, 195 Dongfeng West Road, Guangzhou, 510182 People’s Republic of China Zhe Wang, School of Life Science, Sun Yat-sen University, 135 Xingang West Road, Guangzhou, 510275 People’s Republic of China Ye-Chun Ruan, School of Life Science, Sun Yat-sen University, 135 Xingang West Road, Guangzhou, 510275 People’s Republic of China Wen-Liang Zhou, School of Life Science, Sun Yat-sen University, 135 Xingang West Road, Guangzhou, 510275 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The aim of the study was to identify the number and distribution of genotypes of Streptococcus mutans ( S . mutans ) and Streptococcus sobrinus ( S . sobrinus ) isolated from caries-free and caries-active subjects. Non-stimulated saliva, buccal smooth surface of the right upper teeth, fissures of sound occlusal surface, and carious surface were sampled from 7 caries-free and 7 caries-active (DMFT ≥ 6) students aged 22–24 years. S . mutans and S . sobrinus were isolated using Chelex-100 and primarily identified by colony morphology and biochemical characteristics. The isolates of S . mutans were genotyped using arbitrarily primed polymerase chain reaction. A total of 516 isolates of S . mutans were genotyped from 47 sites in 14 students, and 44 different genotypes were determined. All of the caries-free individuals harbored S . mutans but not S . sobrinus , although individuals 3 and 7 had no S . mutans in their saliva. The CFU value of S . mutans on carious surfaces was the highest, and values in saliva, fissures, and occlusal surfaces were higher in caries-active individuals than in caries-free individuals. We detected 28 genotypes of S . mutans in caries-free individuals, each of who carried more than 3 genotypes. However, we found only 16 genotypes of S . mutans in caries-active individuals, each of who carried no more than 3 genotypes. More genotypes are harbored in the saliva, fissures, and smooth surfaces of caries-free individuals than of caries-active individuals. The proportion of samples positive for S . mutans and S . sobrinus was significantly higher in caries-active individuals than in caries-free individuals, and the presence of these species is a risk factor for high DMFT in dental caries. Isolates of S . mutans exist that have apparent genetic diversity. The genotypes of isolates might relate to differences in caries susceptibility. Content Type Journal Article Pages 1-6 DOI 10.1007/s11010-012-1255-5 Authors Qianzhou Jiang, Department of Endodontics, Stomatology Hospital of Guangzhou Medical University, 59 Huangsha Thoroughfare, Guangzhou, China Miao Yu, Department of Endodontics, Stomatology Hospital of Guangzhou Medical University, 59 Huangsha Thoroughfare, Guangzhou, China Zhipeng Min, The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-Most) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, No. 273 Luoyu Road, Wuhan, 430079 China Anhua Yi, The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-Most) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, No. 273 Luoyu Road, Wuhan, 430079 China Dong Chen, The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-Most) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, No. 273 Luoyu Road, Wuhan, 430079 China Qi Zhang, The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-Most) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, No. 273 Luoyu Road, Wuhan, 430079 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    DNA-binding proteins control various cellular processes such as recombination, replication and transcription. This review is aimed to summarize some of the most commonly used techniques to determine DNA–protein interactions. In vitro techniques such as footprinting assays, electrophoretic mobility shift assay, southwestern blotting, yeast one-hybrid assay, phage display and proximity ligation assay have been discussed. The highly versatile in vivo techniques such as chromatin immunoprecipitation and its variants, DNA adenine methyl transferase identification as well as 3C and chip-loop assay have also been summarized. In addition, some in silico tools have been reviewed to provide computational basis for determining DNA–protein interactions. Biophysical techniques like fluorescence resonance energy transfer (FRET) techniques, FRET–FLIM, circular dichroism, atomic force microscopy, nuclear magnetic resonance, surface plasmon resonance, etc. have also been highlighted. Content Type Journal Article Pages 1-21 DOI 10.1007/s11010-012-1269-z Authors Bipasha Dey, Department of Biotechnology, Jaypee Institute of Information Technology, A-10 Sector-62, Noida, 201307 Uttar Pradesh, India Sameer Thukral, Department of Biotechnology, Jaypee Institute of Information Technology, A-10 Sector-62, Noida, 201307 Uttar Pradesh, India Shruti Krishnan, Department of Biotechnology, Jaypee Institute of Information Technology, A-10 Sector-62, Noida, 201307 Uttar Pradesh, India Mainak Chakrobarty, Department of Biotechnology, Jaypee Institute of Information Technology, A-10 Sector-62, Noida, 201307 Uttar Pradesh, India Sahil Gupta, Department of Biotechnology, Jaypee Institute of Information Technology, A-10 Sector-62, Noida, 201307 Uttar Pradesh, India Chanchal Manghani, Department of Biotechnology, Jaypee Institute of Information Technology, A-10 Sector-62, Noida, 201307 Uttar Pradesh, India Vibha Rani, Department of Biotechnology, Jaypee Institute of Information Technology, A-10 Sector-62, Noida, 201307 Uttar Pradesh, India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Chondroid cell from human adipose-derived stem cells (ADSCs) has emerged as an alternative treatment option for articular cartilage defects. Herein, we successfully compared ADSCs, normal chondrocytes, and chondroid cells. The comparative study of ADSCs and chondroid cells revealed type II collagen (COL II) and glycosaminoglycans expression of chondroid cells were similar to those in normal chondrocytes, and much higher than ADSCs. Using atomic force microscope (AFM) and laser confocal scanning microscopy (LCSM), we compared the differences in morphology, mechanical properties, and F-actin distribution between chondroid cells and normal chondrocytes. Our results showed no differences observed between these two types of cells regarding morphology, stiffness, and F-actin distribution. However, found that the adhesion force in chondroid cells was lower than that in normal chondrocytes. Taken together, our AFM and LCSM analyses suggest that the lower adhesion force in chondroid cells may contribute to the dedifferentiation of ADSC-derived chondroid cells. Future examination of surface adhesion force-related protein expression will likely provide new insight into the molecular mechanisms underlying the dedifferentiation of ADSC-derived chondroid cells. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1263-5 Authors Simin Luo, The First Affiliated Hospital, Jinan University, Guangzhou, 510632 China Qiping Shi, The First Affiliated Hospital, Jinan University, Guangzhou, 510632 China Zhengang Zha, The First Affiliated Hospital, Jinan University, Guangzhou, 510632 China Ping Yao, Department of Medical College, Jinan University, Guangzhou, 510632 China Hongsheng Lin, The First Affiliated Hospital, Jinan University, Guangzhou, 510632 China Ning Liu, The First Affiliated Hospital, Jinan University, Guangzhou, 510632 China Hao Wu, The First Affiliated Hospital, Jinan University, Guangzhou, 510632 China Hua Jin, Department of Chemistry and Institute for Nano-Chemistry, Jinan University, Guangzhou, 510632 China Jiye Cai, Department of Chemistry and Institute for Nano-Chemistry, Jinan University, Guangzhou, 510632 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description: Erratum to: Decitabine-induced apoptosis is derived by Puma and Noxa induction in chronic myeloid leukemia cell line as well as in PBL and is potentiated by SAHA Content Type Journal Article Category Erratum Pages 1-1 DOI 10.1007/s11010-012-1259-1 Authors Barbora Brodská, Institute of Hematology and Blood Transfusion, U Nemocnice 1, 128 20 Prague 2, Czech Republic Petra Otevřelová, Institute of Hematology and Blood Transfusion, U Nemocnice 1, 128 20 Prague 2, Czech Republic Aleš Holoubek, Institute of Hematology and Blood Transfusion, U Nemocnice 1, 128 20 Prague 2, Czech Republic Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Osteosarcoma (OS) severely threatens the health of young people and understanding on the molecular mechanisms of OS etiology enables gene therapy to become an effective therapeutic modality. However, insufficient expression level of genes using existing vectors limits the clinical application of gene therapy for OS. To solve the problem, we developed an oncolytic adenoviral vector, OAT, which can selectively and efficiently replicate in OS cells to enhance the expression of transferred genes. We demonstrated that OAT-mediated TRAIL expression is significantly elevated after infection of OS cells than replication-incompetent Ad5 vector. Increased antitumor capacity was observed in OS cells after OAT-TRAIL treatment both in vitro and in vivo. In normal cells, adenoviral replication, TRAIL expression and growth-inhibiting effect were quite limited when OAT-TRAIL was administrated, showing a high biosafety of this oncolytic adenoviral vector. Collectively, we generated an efficient and promising expression vector for OS gene therapy. Content Type Journal Article Pages 337-344 DOI 10.1007/s11010-012-1235-9 Authors Li Chunbao, Department of Orthopaedic Surgery, General Hospital of People’s Liberation Army, Beijing, 100853 China Cheng Qianpeng, Department of Endocrinology and Metabolism, General Hospital of Beijing Military Region, Beijing, 100700 China Liu Jia, Institute of Oceanology, Chinese Academy of Science, Qingdao, 266071 China Wang Bin, Gastroenterology Institute of Surgery Research, Daping Hospital, The Third Military Medical University, Chongqing, 400042 China Chen Dongfeng, Gastroenterology Institute of Surgery Research, Daping Hospital, The Third Military Medical University, Chongqing, 400042 China Liu Yujie, Department of Orthopaedic Surgery, General Hospital of People’s Liberation Army, Beijing, 100853 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177 Journal Volume Volume 364 Journal Issue Volume 364, Numbers 1-2

Description:    Chlorinated persistent organic pollutants, commonly referred to as organochlorine compounds (OCs), are chemicals of environmental concern that were mostly used historically as pesticides, solvents, flame retardants, and other applications, though some still continue to be produced. OCs accumulate in adipose tissue because of their hydrophobicity. Evidence suggests that OCs modulate adipose tissue metabolism and could affect its development. At the cellular level, the development of adipose tissue is partly controlled by replication of preadipocytes, which may in turn be modulated by contaminants. The aim of this study was to determine whether exposure to specific OCs and to different concentrations, some mimecting those achieved in human tissues that are exposed to chemicals from the environment, affects human preadipocyte proliferation capacity. Human preadipocytes were exposed to various concentrations (3–500 μM) of highly prevalent OCs (PCBs 77, 153 and DDE) for 48 h. At the end of the exposure period, quantification of cell density was assessed by a cell proliferation ELISA assay. Preadipocyte proliferation significantly increased (~28–72%) in response to most of the concentrations of PCB 153 and DDE as compared to the control. These findings suggest that exposure to some OCs and concentrations increase the proliferative capacity of human preadipocytes. Content Type Journal Article Pages 1-4 DOI 10.1007/s11010-012-1268-0 Authors Natalie A. Chapados, Montfort Hospital Research Institute, Nutrition and Metabolism Research Unit (NMRU), Montfort Hospital, 713 Montreal Road, Pavilion E, 1st Floor, room 2E132, Ottawa, ON K1K 0T2, Canada Claudia Casimiro, Faculty of Health Sciences, University of Ottawa, Ottawa, ON, Canada Michael A. Robidoux, Indigenous Health Group, University of Ottawa, Ottawa, ON, Canada François Haman, Montfort Hospital Research Institute, Nutrition and Metabolism Research Unit (NMRU), Montfort Hospital, 713 Montreal Road, Pavilion E, 1st Floor, room 2E132, Ottawa, ON K1K 0T2, Canada Malek Batal, Montfort Hospital Research Institute, Nutrition and Metabolism Research Unit (NMRU), Montfort Hospital, 713 Montreal Road, Pavilion E, 1st Floor, room 2E132, Ottawa, ON K1K 0T2, Canada Jules M. Blais, Department of Biology, University of Ottawa, Ottawa, ON, Canada Pascal Imbeault, Montfort Hospital Research Institute, Nutrition and Metabolism Research Unit (NMRU), Montfort Hospital, 713 Montreal Road, Pavilion E, 1st Floor, room 2E132, Ottawa, ON K1K 0T2, Canada Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Epstein–Barr virus (EBV) is a ubiquitous human herpesvirus implicated in lymphomas, such as Burkitt’s lymphoma, Hodgkin’s lymphoma, and NK/T cell lymphoma. MicroRNAs (miRNAs) are 19–25 nucleotide long single-stranded RNAs involved in post-transcriptional gene regulation. miRNAs are mainly transcribed by RNA polymerase II (pol II) to have stem–loop structures and subsequently processed by Drosha and Dicer. EBV miRNAs are expressed in B cells, nasopharyngeal carcinoma cells, and gastric carcinoma cells infected with EBV. EBV miRNAs can be divided into two groups: BHRF1 miRNAs and BART miRNAs. In this study, we investigated the biogenesis of EBV miRNAs. Treatment of the SNU-719 EBV-positive gastric cancer cell line with α-amanitin at a concentration that selectively inhibits RNA polymerase II activity decreased the expression levels of BART miRNAs. The expression levels of BART miRNAs were also reduced by RNA interference targeting Drosha and Dicer. Two of each C/EBPβ and c-Myc binding sites are located upstream of the proposed initiation sites for primary BART miRNA transcripts. Knockdown of C/EBPβ but not c-Myc using siRNAs reduced BART miRNA expression by 25–55% compared with the control. These results suggest that BART miRNAs are transcribed by pol II and undergo a similar biogenesis process with cellular miRNAs. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1261-7 Authors Do Nyun Kim, Department of Medical Lifescience, Research Institute of Immunobiology, College of Medicine, The Catholic University of Korea, 505 Banpo-dong, Seocho-gu, Seoul, 137-701 Republic of Korea Suk Kyeong Lee, Department of Medical Lifescience, Research Institute of Immunobiology, College of Medicine, The Catholic University of Korea, 505 Banpo-dong, Seocho-gu, Seoul, 137-701 Republic of Korea Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Currently, obesity is an important health problem in all countries, both developed and developing. Dietary habits and neurohormonal imbalances play a critical role in obesity. Circulating amounts of ghrelin, which is a neurohormonal hormone, decrease with obesity and increase with weight loss. Although it is known that both mRNA and peptide version of the ghrelin hormone are expressed in almost all tissues of both humans and animals, it is not known how obesity changes the expression of this hormone in the tissues, with the exception of the gastrointestinal system tissues. Therefore, the objective of the present study is to show how diet-induced obesity in rats changes ghrelin expression in all system tissues, and thus, to shed light on the etiopathology of obesity. The study included 12 male and 12 female 2-month-old Wistar albino species rats. The animals in the control group were fed on standard rat pellet, while those in the experiment group were fed ad libitum on a cafeteria-style diet for 2 months. When their body mass index reached 1 g/cm 2 , diet-induced obese (DIO) rats were sacrificed in a sterile environment after one night fasting. Ghrelin localizations in the tissues were studied immunohistochemically using avidin-biotin-peroxidase complex (ABC) method, while tissue ghrelin amounts were analyzed using radioimmunoassay (RIA) method. When the ghrelin amounts in the urogenital system (with the exception of kidney tissues), sensory organs, respiratory system, immune system, skeletal muscle system, cardiovascular system, nervous system, and adipose tissue of rats analyzed by RIA method were compared to those in the control group, tissue ghrelin amounts in the DIO group were found lower. Immunohistochemical findings which showed a similar fall in ghrelin concentrations in the tissues were parallel to RIA results. In addition, ghrelin was shown to be synthesized in the cardiovascular system, heart muscle cells, tails of the sperms, hair follicles, lacrimal glands, tongue, and teeth of rats for the first time in this study and ghrelin syntheses in these tissues were found to decrease in obesity. Nutritional obesity is among the most common causes of obesity and the findings we have obtained through diet-induced obesity will contribute to the illumination of the etiopathology of obesity. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1256-4 Authors Suleyman Aydin, Department of Medical Biochemistry and Clinical Biochemistry (Firat Hormones Research Group), School of Medicine, Firat University Hospital, Firat University, Elazig, 23119 Turkey İbrahim Sahin, Department of Medical Biochemistry and Clinical Biochemistry (Firat Hormones Research Group), School of Medicine, Firat University Hospital, Firat University, Elazig, 23119 Turkey Yusuf Ozkan, Department of Internal Medicine, Firat University Hospital, Firat University, Elazig, 23119 Turkey Ersel Dag, Department of Neurology, School of Medicine, Kirikkale University, Kirikkale, 71100 Turkey Ahmet Gunay, Department of Periodontics, Faculty of Dentistry, Dicle University, Diyarbakir, 21280 Turkey Saadet Pilten Guzel, Department of Medical Biochemistry and Clinical Biochemistry (Firat Hormones Research Group), School of Medicine, Firat University Hospital, Firat University, Elazig, 23119 Turkey Zekiye Catak, Department of Medical Biochemistry and Clinical Biochemistry (Firat Hormones Research Group), School of Medicine, Firat University Hospital, Firat University, Elazig, 23119 Turkey Mehmet Resat Ozercan, Department of Medical Pathology, Firat University Hospital, Firat University, Elazig, 23119 Turkey Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Hypertrophic scarring (HTS) is a common fibroproliferative disorder that typically follows thermal and other injuries involving the deep dermis. These pathogenic mechanisms are regulated by connective tissue growth factor (CTGF) and transforming growth factor-β. We found that neuregulin-1 (NRG1), as well as NRG receptors, HER-2, and HER-3 were upregulated in HTS fibroblasts (HTSF), compared with normal fibroblasts. Furthermore, NRG1 stimulation increased the expression of CTGF in HTSF. In the presence of inhibitors of PI3K, Src, Smad, or reactive oxygen species, the effect of NRG1 on CTGF expression decreased significantly. In particular, the combination of LY294002 or PP2 with SB431542 blocked NRG1-mediated CTGF expression in HTSF. Finally, we demonstrated that siRNA for CTGF, AG825, LY294002, and PP2, either alone or in co-treatment, effectively reduced extracellular matrix expression. Taken together, our results suggest that NRG1 is involved in fibrotic scar pathogenesis via PI3K- or Src-mediated CTGF expression. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1258-2 Authors Jun-Sub Kim, Specialization Research Center, Hallym University Burn Institute, Hangang Sacred Heart Hospital, Hallym University, Seoul, South Korea Ihn-Geun Choi, Department of Neuropsychiatry, Hallym University Burn Institute, Hangang Sacred Heart Hospital, Hallym University, Seoul, South Korea Boung-Chul Lee, Department of Neuropsychiatry, Hallym University Burn Institute, Hangang Sacred Heart Hospital, Hallym University, Seoul, South Korea Jae-Bong Park, Department of Biochemistry, Hallym University Burn Institute, College of Medicine, Hallym University, Chuncheon, Kangwon-Do, South Korea Jin-Hee Kim, Specialization Research Center, Hallym University Burn Institute, Hangang Sacred Heart Hospital, Hallym University, Seoul, South Korea Je Hoon Jeong, Department of Neurological Surgery, Hallym University Burn Institute, Hangang Sacred Heart Hospital, Hallym University, Seoul, South Korea Ji Hoon Jeong, College of Pharmacy, Sungkyunkwan University, Suwon, South Korea Cheong Hoon Seo, Department of Rehabilitation Medicine, Hangang Sacred Heart Hospital, Hallym University, 94-200 Yeongdeungpo-Dong, Yeongdeungpo-Gu, Seoul, 150-719 South Korea Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The G protein-coupled oestrogen receptor GPER1, also known as GPR30, has been implicated in oestrogen signalling, but the physiological importance of GPER1 is not fully understood. The GPER1 agonist G-1 has become an important tool to assess GPER1-mediated cellular effects. Here, we report that this substance, besides acting via GPER1, affects the microtubule network in endothelial cells. Treatment with G-1 (3 μM) for 24 h reduced DNA synthesis by about 60 % in mouse microvascular endothelial bEnd.3 cells. Treatment with 3 μM G-1 prevented outgrowth of primary endothelial cells from mouse aortic explants embedded in Matrigel. Treatment with G-1 (0.3–3 μM) for 24 h disrupted bEnd.3 cell and HUVEC microtubule structure in a concentration-dependent manner as assessed by laser-scanning confocal immunofluorescence microscopy. G-1-induced (3 μM) disruption of microtubule was observed also after acute (3 and 6 h) treatment and in the presence of the protein synthesis inhibitor cycloheximide. Disruption of microtubules by 3 μM G-1 was observed in aortic smooth muscle cells obtained from both GPER1 knockout and wild-type mice, suggesting that G-1 influences microtubules through a mechanism independent of GPER1. G-1 dose dependently (10–50 μM) stimulated microtubule assembly in vitro. On the other hand, microtubules appeared normal in the presence of 10–50 μM G-1 as determined by electron microscopy. We suggest that G-1-promoted endothelial cell anti-proliferation is due in part to alteration of microtubule organization through a mechanism independent of GPER1. This G-1-promoted mechanism may be used to block unwanted endothelial cell proliferation and angiogenesis such as that observed in, e.g. cancer. Content Type Journal Article Pages 1-11 DOI 10.1007/s11010-012-1301-3 Authors Anders Holm, Department of Experimental Medical Science, Lund University, BMC D12, 221 84 Lund, Sweden Per-Olof Grände, Department of Clinical Science, Lund University and Lund University Hospital, BMC F11, 221 84 Lund, Sweden Richard F. Ludueña, Department of Biochemistry, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900, USA Björn Olde, Department of Experimental Medical Science, Lund University, BMC D12, 221 84 Lund, Sweden Veena Prasad, Department of Biochemistry, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900, USA L. M. Fredrik Leeb-Lundberg, Department of Experimental Medical Science, Lund University, BMC D12, 221 84 Lund, Sweden Bengt-Olof Nilsson, Department of Experimental Medical Science, Lund University, BMC D12, 221 84 Lund, Sweden Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The study was focused to the influence of higher intake of cholesterol on properties of the renal Na,K-ATPase, a key system in maintaining the homeostasis of sodium in the organism. Feeding for 4 weeks with cholesterol-enriched food for rats afflicted with hereditary hypertriglyceridemia by itself enhanced the activity of Na,K-ATPase, probably as a consequence of higher number of active enzyme molecules as suggested by 32 % increase of V max value. This may be hypothesized as a reason for the increased retention of sodium. Three-week-lasting treatment of animals kept on high cholesterol diet with antioxidant SMe1EC2 in a dose of 10 mg kg −1  day −1 normalized the function of renal Na,K-ATPase to the level comparable in hypertriglyceridemic rats fed with the standard diet. Therefore, our results suggest that the antioxidant SMe1EC2 in the applied dose seems to be effective in the attenuation of cholesterol-induced retention of sodium. Treatment for 3 weeks with Fenofibrate in a dose of 100 mg kg −1  day −1 reversed the function of renal Na,K-ATPase only slightly. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1281-3 Authors Lucia Mézešová, Institute for Heart Research, Department of Biochemistry, Slovak Academy of Sciences, Dúbravská cesta 9, P.O. Box 104, 840 05 Bratislava 45, Slovak Republic Veronika Jendruchová-Javorková, Institute for Heart Research, Department of Biochemistry, Slovak Academy of Sciences, Dúbravská cesta 9, P.O. Box 104, 840 05 Bratislava 45, Slovak Republic Jana Vlkovičová, Institute for Heart Research, Department of Biochemistry, Slovak Academy of Sciences, Dúbravská cesta 9, P.O. Box 104, 840 05 Bratislava 45, Slovak Republic Zuzana Kyselova, Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Dúbravská cesta 9, 841 04 Bratislava, Slovak Republic Jana Navarová, Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Dúbravská cesta 9, 841 04 Bratislava, Slovak Republic Štefan Bezek, Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Dúbravská cesta 9, 841 04 Bratislava, Slovak Republic Norbert Vrbjar, Institute for Heart Research, Department of Biochemistry, Slovak Academy of Sciences, Dúbravská cesta 9, P.O. Box 104, 840 05 Bratislava 45, Slovak Republic Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Previously, we reported that fructose-1,6-bisphosphate (FBP) was taken up by rat cardiac myocytes by two processes: a component that was saturable at micromolar levels and a nonsaturable component that dominated at millimolar levels. Here, we continued to characterize the saturable high-affinity component, with the aim of identifying the physiological substrate and role for this activity. ATP, ADP, and AMP inhibited the uptake of FBP with apparent affinities of 0.2–0.5 mM. Fumarate and succinate were very weak inhibitors. Several phosphorylated sugars (ribulose-1,5-phosphate, fructose-1-phosphate, ribose-5-phosphate, and inositol-2-phosphate) inhibited FBP uptake with apparent affinities of 40–500 μM. As in our previous study, no tested compound appeared to bind as well as FBP. The data suggest that the best ligands have two phosphoryl groups separated by at least 8 Å. The rates of FBP uptake were measured from 3° to 37°. The calculated activation energy was 15–50 kJ/mol, similar to other membrane transport processes. Uptake of FBP was tested in several types of cells other than cardiac myocytes, and compared to the uptake of 2-deoxyglucose and l -glucose. While FBP uptake in excess of that of l -glucose was observed in some cells, in no case was the uptake as high as in cardiac myocytes. The physiological substrate and role for the high-affinity FBP uptake activity remain unknown. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1279-x Authors Thomas J. Wheeler, Department of Biochemistry and Molecular Biology, University of Louisville School of Medicine, Louisville, KY, USA Sufan Chien, Department of Surgery, University of Louisville School of Medicine, Health Sciences Center, Louisville, KY 40202, USA Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The metastasis of colorectal cancer is one of the most common causes of death in the world. In this investigation, we used the human colon cancer cell lines LOVO and HT29 as model systems to determine the role of the chloride intracellular channel 1 (CLIC1) in the metastasis of colonic cancer. In the present study, we found that regulatory volume decrease (RVD) capacity was markedly up-regulated in LOVO cells, which are characterized by a high metastatic potential. Functionally suppressing CLIC1 using the specific chloride intracellular channel 1 blocker Indanyloxyacetic acid 94 inhibited RVD and decreased the migration and invasion of colon cancer cells. Moreover, these effects occurred in a dose-dependent manner. The migration and invasion abilities in two cell lines also were inhibited by the knockdown of CLIC1 using small interfering RNA transfection. The mRNA and protein expression of CLIC1 is up-regulated in LOVO cells. In human colon cancer cells, CLIC1 is primarily located in the plasma membrane, where it functions as a chloride channel. Taken together, the results suggest that CLIC1 modulates the metastasis of colon cancer through its RVD-mediating chloride channel function. This study demonstrates, for the first time, that CLIC1 regulates the migration and invasion of colon cancer. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1271-5 Authors Pan Wang, Department of General Surgery and Minimally Invasive Gastrointestinal Surgery, Southwest Hospital, Third Military Medical University, 29 Gaotanyan Street, Shapingba District, Chong Qing, 400038 People’s Republic of China Chao Zhang, Department of General Surgery and Minimally Invasive Gastrointestinal Surgery, Southwest Hospital, Third Military Medical University, 29 Gaotanyan Street, Shapingba District, Chong Qing, 400038 People’s Republic of China PeiWu Yu, Department of General Surgery and Minimally Invasive Gastrointestinal Surgery, Southwest Hospital, Third Military Medical University, 29 Gaotanyan Street, Shapingba District, Chong Qing, 400038 People’s Republic of China Bo Tang, Department of General Surgery and Minimally Invasive Gastrointestinal Surgery, Southwest Hospital, Third Military Medical University, 29 Gaotanyan Street, Shapingba District, Chong Qing, 400038 People’s Republic of China Tao Liu, Department of General Surgery and Minimally Invasive Gastrointestinal Surgery, Southwest Hospital, Third Military Medical University, 29 Gaotanyan Street, Shapingba District, Chong Qing, 400038 People’s Republic of China Hao Cui, Department of General Surgery and Minimally Invasive Gastrointestinal Surgery, Southwest Hospital, Third Military Medical University, 29 Gaotanyan Street, Shapingba District, Chong Qing, 400038 People’s Republic of China JianHua Xu, Department of General Surgery and Minimally Invasive Gastrointestinal Surgery, Southwest Hospital, Third Military Medical University, 29 Gaotanyan Street, Shapingba District, Chong Qing, 400038 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Reactive oxygen species has been suggested to be one of the key factors associated with the development of obesity. During spontaneous differentiation of mouse stromal preadipocytes OP9 into adipocytes, intracellular superoxide anion radicals (O 2 −. ) level markedly increases and is accompanied by a significant elevation of intracellular lipid accumulation. This differentiation-dependent increase in intracellular O 2 −. level positively correlated with the intracellular augmentation of the lipid level. Super-highly hydroxylated fullerene (SHH-F; C 60 (OH) 44 ), a novel polyhydroxylated fullerene derivative, quenched intracellular O 2 −. , and lipid accumulation to 38.7 and 42.7 % of that in the control, respectively. By thin-layer chromatographic analysis of extracted cellular lipid components, SHH-F clearly decreased the triglycerides ratio in the whole lipid droplet fraction, but scarcely influenced other lipids components. PPARγ2 expression, which plays a key role in regulating adipogenic differentiation, was significantly suppressed by SHH-F at the late stage of differentiation, with unaltered PPARγ1 expression. The intracellular superoxide anion radical augmentation preceded expression of PPARγ2, strongly suggesting that the primary O 2 −. generation was closely associated with lipid accumulation and subsequent PPARγ2 induction. These results indicate that SHH-F suppresses intracellular lipid accumulation, particularly in lipid droplets, and decreases O 2 −. level and subsequent PPARγ2 upregulation during spontaneous differentiation of OP9 preadipocytes into adipocytes. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1297-8 Authors Yasukazu Saitoh, Laboratory of Cell-Death Control BioTechnology, Faculty of Life and Environmental Sciences, Prefectural University of Hiroshima, 562 Nanatsuka, Shobara, Hiroshima, 727-0023 Japan Hiromi Mizuno, Laboratory of Cell-Death Control BioTechnology, Faculty of Life and Environmental Sciences, Prefectural University of Hiroshima, 562 Nanatsuka, Shobara, Hiroshima, 727-0023 Japan Li Xiao, Laboratory of Cell-Death Control BioTechnology, Faculty of Life and Environmental Sciences, Prefectural University of Hiroshima, 562 Nanatsuka, Shobara, Hiroshima, 727-0023 Japan Sayuri Hyoudou, Vitamin C60 BioResearch Corporation, 9F, 1-3-19 Yaesu Chuo-ku, Tokyo, 103-0028 Japan Ken Kokubo, Division of Applied Chemistry, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871 Japan Nobuhiko Miwa, Laboratory of Cell-Death Control BioTechnology, Faculty of Life and Environmental Sciences, Prefectural University of Hiroshima, 562 Nanatsuka, Shobara, Hiroshima, 727-0023 Japan Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    PPARδ, a member of the peroxisome proliferator-activated receptor superfamily, plays a key role in the transcriptional regulation of genes involved in cellular lipid and energy metabolism. Therefore, PPARδ may represent a new target for the treatment of obesity, hyperlipidemia, and type 2 diabetes. MafA is a β-cell-specific and glucose-regulated transcriptional activator for insulin gene expression and plays a crucial role in pancreas development, β-cell differentiation as well as maintenance of β-cell function. However, little is known about how PPARδ regulates MafA and ameliorates glucose-stimulated insulin secretion impaired by free fatty acids (FFA). In the present study, we evaluated the basal insulin secretion (BIS), glucose-stimulated insulin secretion (GSIS), and insulin secretion index (ISI) of INS-1E cells that were cultured in media supplemented with or without 0.5 mM palmitate and treated with or without a PPARδ agonist (GW501516) or PPARδ siRNA. The expression of MafA, glucose transportor-2 (GLUT2), and insulin was found to be up-regulated in cells treated with GW501516. Finally, analysis of the level of JNK phosphorylation revealed that activated PPARδ could inhibit the activation of JNK and increase the expression of MafA. Accordingly, the insulin secretion dysfunction in lipotoxic INS-1E cells was improved. Collectively, these results demonstrate that activation of PPARδ improves insulin secretion impaired by palmitate and plays a role in the JNK–MafA–GLUT2 pathway. Content Type Journal Article Pages 1-7 DOI 10.1007/s11010-012-1296-9 Authors Mingming Cao, Division of Endocrinology, West China Hospital of Sichuan University, 37 Guoxuexiang, Chengdu, 610041 Sichuan, China Yang Long, Research Laboratory of Endocrine and Metabolic Diseases, West China Hospital of Sichuan University, Chengdu, China Yuzhen Tong, School of Clinical Medicine, West China Hospital of Sichuan University, 37 Guoxuexiang, Chengdu, 610041 Sichuan, China Jun Wan, Division of Endocrinology, West China Hospital of Sichuan University, 37 Guoxuexiang, Chengdu, 610041 Sichuan, China Nanwei Tong, Division of Endocrinology, West China Hospital of Sichuan University, 37 Guoxuexiang, Chengdu, 610041 Sichuan, China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    GPR12, a member of the orphan G-protein-coupled receptor family, constitutively activates the Gs protein and increases intracellular cyclic AMP concentrations. GPR12 can be activated by its known ligand—sphingosylphosphorylcholine, which regulates cellular physiological activities, including proliferation, neurite extension, cell clustering, and maintenance of meiotic arrest. However, signaling pathways involved in the GPR12-mediated physiological and biochemical changes are still not clearly illustrated. In the present study, heterologous GPR12 expression was demonstrated to promote proliferation and survival in human embryonic kidney 293 cells. Immunochemical analysis showed that Ki67, a prototypic cell cycle-related nuclear protein, might participate in the regulation of GPR12-mediated cell proliferation. Activation of extracellular signal-regulated protein kinase signaling and increased total Erk1/2 and B-cell lymphoma/leukemia-2 expression were also observed in HEK293 cells overexpressing human GPR12. In addition, we found that GPR12 promoted cell survival under serum deprivation, indicating that GPR12 may play a role in cell proliferation and survival. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1287-x Authors Xiaoming Lu, Institutes for Advanced Interdisciplinary Research, East China Normal University, 3663 North Zhongshan Road, Shanghai, 200062 China Ning Zhang, Institutes for Advanced Interdisciplinary Research, East China Normal University, 3663 North Zhongshan Road, Shanghai, 200062 China Bo Meng, Key Laboratory of Brain Functional Genomics, MOE & STCSM, East China Normal University, 3663 North Zhongshan Road, Shanghai, 200062 China Suzhen Dong, Key Laboratory of Brain Functional Genomics, MOE & STCSM, East China Normal University, 3663 North Zhongshan Road, Shanghai, 200062 China Yinghe Hu, Institutes for Advanced Interdisciplinary Research, East China Normal University, 3663 North Zhongshan Road, Shanghai, 200062 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Both the aorta and the liver are major organs that play important roles in lipid metabolism, and they are subject to systemic as well as local inflammatory responses in metabolic syndrome. Our previous study indicated that TNFα deficiency influences atherogenesis by reducing inflammation of the aorta. To better understand this phenomenon, the mRNA and protein expression profiles of a panel of cytokines in the aorta and liver of young TNFα-null (TNFα −/− ) mice were analyzed and compared with age- and gender-matched wild-type (WT) control mice. In the aorta, IL-2 and GM-CSF were up-regulated versus WT mice, while IL-1β, IL-4, IL-6, IL-10, MCP-1, IFN-γ, and the adhesion molecules ICAM-1 and VCAM-1 were down-regulated. In the liver, however, the expressions of NF-κB, IL-1, IL-2, IL-6, IL-10, ICAM-1, and VCAM-1 were significantly up-regulated in TNFα −/− mice, while IFN-γ and IL-4 were down-regulated. Out of the 62 cytokines analyzed, 22 in the aorta and 27 in the liver were altered by 2–fivefolds at the protein level in TNFα −/− mice. Our data demonstrated that the loss of TNFα function led to various changes in the levels of cytokine expression in these organs at both the transcriptional and translational levels. These results indicated that the changes in cytokine expression patterns in the aorta and the liver may further influence the progression of systemic or local lipid metabolism dysregulation and pathogenesis in animals with TNFα dysfunction representing inflammation-related diseases, such as atherosclerosis and metabolic syndrome. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1283-1 Authors Yanrong Sun, The Key Laboratory of Animal Resistant Biology of Shandong, College of Life Sciences, Shandong Normal University, #88 East Wenhua Ave., Jinan, 250014 China Miao Yin, The Key Laboratory of Animal Resistant Biology of Shandong, College of Life Sciences, Shandong Normal University, #88 East Wenhua Ave., Jinan, 250014 China Liang Zhang, The Key Laboratory of Animal Resistant Biology of Shandong, College of Life Sciences, Shandong Normal University, #88 East Wenhua Ave., Jinan, 250014 China Jie Pan, The Key Laboratory of Animal Resistant Biology of Shandong, College of Life Sciences, Shandong Normal University, #88 East Wenhua Ave., Jinan, 250014 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The aim of this study was to evaluate whether citreorosein (CIT), a naturally occurring anthraquinone isolated from Polygoni cuspidati ( P. cuspidati ) radix, modulates degranulation and 5-lipoxygenase (5-LO)-dependent leukotriene C 4 (LTC 4 ) generation in mast cells. Cit suppresses both degranulation and the generation of LTC 4 in a dose-dependent manner in stem cell factor (SCF)-mediated mouse bone marrow-derived mast cells (BMMCs). With regard to its molecular mechanism of action, we investigated the effects of CIT on intracellular signaling and mast cell activation employing BMMCs. Binding of SCF to c-Kit on mast cell membranes induced increases in intrinsic tyrosine kinase Syk activity and activation of multiple downstream events including phosphorylation of phospholipase Cγ (PLCγ), mobilization of intracellular Ca 2+ , phosphatidylinositol 3-kinase (PI3K), Akt, MAP kinases (MAPKs), translocation of phospho-phospholipase A 2 (PLA 2 ) and 5-LO. The results from the biochemical analysis demonstrate that CIT attenuates degranulation and LTC 4 generation through the suppression of multiple step signaling and would be beneficial for the prevention of allergic inflammation. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1273-3 Authors Yue Lu, College of Pharmacy, Yeungnam University, Gyeongsan, 712-749 Republic of Korea Ying Li, College of Pharmacy, Yeungnam University, Gyeongsan, 712-749 Republic of Korea Yurndong Jahng, College of Pharmacy, Yeungnam University, Gyeongsan, 712-749 Republic of Korea Jong-Keun Son, College of Pharmacy, Yeungnam University, Gyeongsan, 712-749 Republic of Korea Hyeun Wook Chang, College of Pharmacy, Yeungnam University, Gyeongsan, 712-749 Republic of Korea Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Ginsenoside Rg1 promotes antioxidative protection and intracellular calcium homeostasis in cardiomyocytes hypoxia/reoxygenation (H/R) model. However, the pharmacological effects of G-Rg1 on autophagy in cardiomyocytes have not been reported. In this study, we employed H9c2 cardiomyocytes as a model to investigate the effects of G-Rg1 on autophagy in cardiomyocytes under H/R stress. Our results showed that H/R induced increased level of LC3B-2, an autophagy marker, in a time-dependent manner in association with decreased cell viability and cellular ATP content. H/R-induced autophagy and apoptosis were further confirmed by morphological examination. 100 μmol/l Rg1-inhibited H/R induced autophagy and apoptosis, and this was associated with the increase of cellular ATP content and the relief of oxidative stress in the cells. Mechanistically, we found that Rg1 inhibited the activation of AMPKα, promoted the activation of mTOR, and decreased the levels of LC3B-2 and Beclin-1. In conclusion, our data suggest that H/R induces autophagy in H9c2 cells leading to cell injury. Rg1 inhibits autophagosomal formation and apoptosis in the cells, which may be beneficial to the survival of cardiomyocytes under H/R. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1265-3 Authors Zi-Long Zhang, Department of Geriatric, Peking University First Hospital, No. 8, Xishiku Street, Xicheng District, Beijing, 100034 China Yan Fan, Department of Geriatric, Peking University First Hospital, No. 8, Xishiku Street, Xicheng District, Beijing, 100034 China Mei-Lin Liu, Department of Geriatric, Peking University First Hospital, No. 8, Xishiku Street, Xicheng District, Beijing, 100034 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Lamins are the major structural components of the nucleus and mutations in the human lamin A gene cause a number of genetic diseases collectively termed laminopathies. At the cellular level, lamin A mutations cause aberrant nuclear morphology and defects in nuclear functions such as the response to DNA damage. We have investigated the mechanism of depletion of a key damage sensor, ATR (Ataxia-telangiectasia-mutated-and-Rad3-related) kinase, in HeLa cells expressing lamin A mutants or lamin A shRNA. The degradation of ATR kinase in these cells was through the proteasomal pathway as it was reversed by the proteasomal inhibitor MG132. Expression of lamin A mutants or shRNA led to transcriptional activation of three ubiquitin ligase components, namely, RNF123 (ring finger protein 123), HECW2 (HECT domain ligase W2) and the F-box protein FBXW10. Ectopic expression of RNF123, HECW2 or FBXW10 directly resulted in proteasomal degradation of ATR kinase and the ring domain of RNF123 was required for this degradation. However, these ligases did not alter the stability of DNA-dependent protein kinase, which is not depleted upon lamin misexpression. Although degradation of ATR kinase was reversed by MG132, it was not affected by the nuclear export inhibitor, leptomycin B, suggesting that ATR kinase is degraded within the nucleus. Our findings indicate that lamin misexpression can lead to deleterious effects on the stability of the key DNA damage sensor, ATR kinase by upregulation of specific components of the ubiquitination pathway. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1272-4 Authors Bhattiprolu Muralikrishna, Centre for Cellular and Molecular Biology (CSIR), Uppal Road, Hyderabad, 500007 India Pankaj Chaturvedi, Centre for Cellular and Molecular Biology (CSIR), Uppal Road, Hyderabad, 500007 India Kirti Sinha, Centre for Cellular and Molecular Biology (CSIR), Uppal Road, Hyderabad, 500007 India Veena K. Parnaik, Centre for Cellular and Molecular Biology (CSIR), Uppal Road, Hyderabad, 500007 India Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    ZNF403 , also known as GGNBP2 ( gametogenetin binding protein 2 ), is a highly conserved gene implicated in spermatogenesis. However, the exact biological function of ZNF403 is not clear. In this study, we identified the role of ZNF403 in cell proliferation and cell-cycle regulation by utilizing short hairpin RNA (shRNA)-mediated knockdown. ZNF403-specific shRNA expressing helper-dependent adenoviral vector (HD-Ad-ZNF403-shRNA) was constructed and transduced human cell lines. ZNF403 mRNA and protein expression levels were inhibited as evidenced by real-time PCR and western blot analyses. Noticeably, we found that knockdown of ZNF403 expression suppressed cell proliferation compared to the non-target shRNA and vector controls. Furthermore, cell-cycle analysis demonstrated that downregulation of ZNF403 promoted G2/M cell-cycle arrest in a dose-dependent manner. Moreover, human cell-cycle real-time PCR array revealed that ZNF403 knockdown influenced the expression profile of genes in cell-cycle regulation. Among these genes, western blot analysis confirmed the protein up-regulation of p21 and down-regulation of MCM2 in response to the ZNF403 knockdown. Additionally, knockdown of ZNF403 also showed an anti-carcinogenetic effect on anchorage-independent growth by colony formation assay and tumor cell migration by wound-healing assay with human laryngeal cancer cell line Hep-2 cells. Altogether, our findings suggest an essential role of ZNF403 in cell proliferation and provide a new insight into the function of ZNF403 in regulating the G2/M cell-cycle transition. Content Type Journal Article Pages 1-12 DOI 10.1007/s11010-012-1262-6 Authors Rui Guan, Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital and Cancer Research Institute, Central South University, Changsha, 410008 Hunan Province, China Xiao-Yan Wen, Keenan Research Center, Li Ka Shing Knowledge Institute, St. Michael’s Hospital and Department of Medicine, University of Toronto, Toronto, ON, Canada Jing Wu, Physiology and Experimental Medicine Program, Hospital for Sick Children, 555 University Avenue, Toronto, ON M5G 1X8, Canada Rongqi Duan, Physiology and Experimental Medicine Program, Hospital for Sick Children, 555 University Avenue, Toronto, ON M5G 1X8, Canada Huibi Cao, Physiology and Experimental Medicine Program, Hospital for Sick Children, 555 University Avenue, Toronto, ON M5G 1X8, Canada Simon Lam, Physiology and Experimental Medicine Program, Hospital for Sick Children, 555 University Avenue, Toronto, ON M5G 1X8, Canada Defu Hou, Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital and Cancer Research Institute, Central South University, Changsha, 410008 Hunan Province, China Youdong Wang, Keenan Research Center, Li Ka Shing Knowledge Institute, St. Michael’s Hospital and Department of Medicine, University of Toronto, Toronto, ON, Canada Jim Hu, Physiology and Experimental Medicine Program, Hospital for Sick Children, 555 University Avenue, Toronto, ON M5G 1X8, Canada Zhuchu Chen, Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital and Cancer Research Institute, Central South University, Changsha, 410008 Hunan Province, China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    There is a clear relationship between the pelvic floor muscles and urinary systems, which relates to urgency, frequency, incontinence, pelvic pain, and bowel complaints. The specific mechanisms which relate these two systems are not clear. Improved understanding of the relation between the pelvic floor muscles and bladder function is clinically relevant in establishing effective treatments to such problems as incontinence, secondary to birth. The following tissues were collected from normal adult female rabbits: pubococcygeus (Pc) and ischiocavernosus/bulbospongiosus (Ic/Bs) pelvic floor muscles. Bladder body muscle and mucosa, bladder base muscle and mucosa, and leg skeletal muscle were also collected. The following enzymatic assays were performed on each tissue: citrate synthase (CS), sarcoplasmic–endoplasmic reticular ATPase (SERCA), and choline acetyltransferase (ChAT). CS and SERCA activities were significantly higher in the Pc compared with the Ic/Bs pelvic floor muscles, whereas the ChAT activity of the Ic/Bs was higher than that of the Pc muscle. Based on our results, the Pc muscle is expected to have a significantly greater capacity to contract and a higher metabolic activity than those of the Ic/Bs muscles. We believe that an understanding of the biochemical activities of these three biomarker enzymes in normal pelvic floor muscles is essential in evaluating the effects of specific experimental dysfunctions created in pelvic floor muscle activity. Content Type Journal Article Pages 1-5 DOI 10.1007/s11010-012-1347-2 Authors Sara Spettel, Urology, Albany Medical Center, Albany, NY, USA Elise De, Urology, Albany Medical Center, Albany, NY, USA Tamer Elias, Albany College of Pharmacy and Health Sciences, Albany, NY 12208, USA Catherine Schuler, Stratton VA Medical Center, Albany, NY 12208, USA Robert E. Leggett, Stratton VA Medical Center, Albany, NY 12208, USA Robert M. Levin, Stratton VA Medical Center, Albany, NY 12208, USA Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    In type 2 diabetes, pancreatic β-cells cannot secret enough insulin compensate for insulin resistance, which are often accompanied by abnormality in lipid metabolism such as hypertriglyceridemia. It is reported that oxidative stress is involved in pancreatic β-cell dysfunction. However, molecular mechanisms linking between excessive generations of reactive oxygen species (ROS) and β-cell dysfunction and apoptosis induced by high levels of very low-density lipoprotein (VLDL) are poorly understood. In this study, we test the hypothesis that NADPH oxidase 2 (NOX2)-derived ROS may play a key role in dysfunction and apoptosis of pancreatic β-cell induced by VLDL. Our results show that the ApoCIII transgenic mice displayed increased serum TG levels, enhanced generation of ROS and impaired insulin content in pancreatic β-cells. In vitro, the treatment of pancreatic NIT-1 cells with 1 mg/ml VLDL for 12 h stimulated NOX2-derived ROS generation, decreased expression and secretion of insulin. Furthermore, we found that VLDL induced dysfunction and apoptosis of pancreatic β-cells through JNK and p53 pathways, which were rescued by siRNA-mediated NOX2 reduction. In conclusion, our data demonstrate a critical role of NOX2-derived ROS in dysfunction and apoptosis through JNK and p53 pathways in pancreatic β-cells induced by VLDL. Content Type Journal Article Pages 1-11 DOI 10.1007/s11010-012-1402-z Authors Juan Jiao, Graduate School of Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100730, China Lin Dou, Graduate School of Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100730, China Miao Li, The Key Laboratory of Geriatrics, Beijing Hospital & Beijing Institute of Geriatrics, Ministry of Health, Beijing, 100730 China Yonggang Lu, Graduate School of Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100730, China Han-Bang Guo, The Key Laboratory of Geriatrics, Beijing Hospital & Beijing Institute of Geriatrics, Ministry of Health, Beijing, 100730 China Yong Man, The Key Laboratory of Geriatrics, Beijing Hospital & Beijing Institute of Geriatrics, Ministry of Health, Beijing, 100730 China Shu Wang, The Key Laboratory of Geriatrics, Beijing Hospital & Beijing Institute of Geriatrics, Ministry of Health, Beijing, 100730 China Jian Li, Graduate School of Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100730, China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Aging progress and degeneracy of functional activity are mainly attributed to the decreased DNA repair potential. DNA polymerase (pol) δ activity plays an essential role in genome stability by virtue of its crucial DNA replication and repair capacity. To order to clarify the role of DNA pol δ in aging progression, we firstly examined the expressions of its catalytic subunit named DNA pol δ1 in human lymphocytes at different age stages, respectively, and then observed the effect of diseases on DNA pol δ1 in vivo and of nutriture on its expressions in 2BS cells in vitro. Blood samples from the healthy subjects and patients with diabetes mellitus and coronary heart disease were collected, respectively, for analysis of transcription and protein expressions of DNA pol δ1 by RT-PCR and western blot. 2BS cells of PD30 and PD47 were incubated in both normal medium and other mediums of different nutritures for verifying the differential expressions of DNA pol δ1. Results showed that the mRNA expression of DNA pol δ1 decreased substantially with age and the protein levels were well consistent with gene levels. Furthermore, there were no significant differences in DNA pol δ1 expressions between the groups of healthy individuals and the age matched patients. In addition, DNA pol δ1 gene expression levels were not affected by nutritional status in vitro. Our findings collectively confirmed that the down-regulations of DNA pol δ1 are age-related and have little bearing on diseases and nutritures. DNA pol δ1 has great potential for a new biomarker of aging. Content Type Journal Article Pages 1-7 DOI 10.1007/s11010-012-1432-6 Authors Jin-Ling Wang, Department of Clinical Laboratory, Xuanwu Hospital, Capital Medical University, Beijing, 100053 People’s Republic of China Hong-Lin Guo, Department of Clinical Laboratory, Beijing Traditional Chinese Medical Hospital, Capital Medical University, Beijing, 100010 People’s Republic of China Pei-Chang Wang, Department of Clinical Laboratory, Xuanwu Hospital, Capital Medical University, Beijing, 100053 People’s Republic of China Chen-Geng Liu, Department of Clinical Laboratory, Xuanwu Hospital, Capital Medical University, Beijing, 100053 People’s Republic of China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    In this study, we investigated the effect of 6 weeks of swimming training on the ecto-nucleotidase activities and platelet aggregation from rats that developed hypertension in response to oral administration of l -NAME. The rats were divided into four groups: control ( n  = 10), exercise ( n  = 10), l -NAME ( n  = 10), and exercise l -NAME ( n  = 10). The animals were trained five times per week in an adapted swimming system for 60 min with a gradual increase of the workload up to 5 % of animal’s body weight. The results showed an increase in ATP, ADP, AMP, and adenosine hydrolysis, indicating an augment in NTPDase (from 35.3 ± 8.1 to 53.0 ± 15.1 nmol Pi/min/mg protein for ATP; and from 21.7 ± 7.0 to 46.4 ± 15.6 nmol Pi/min/mg protein for ADP as substrate), ecto-5′-nucleotidase (from 8.0 ± 5.7 to 28.1 ± 6.9 nmol Pi/min/mg protein), and ADA (from 0.8 ± 0.5 to 3.9 ± 0.8 U/L) activities in platelets from l -NAME-treated rats when compared to other groups ( p  〈 0.05). A significant augment on platelet aggregation in l -NAME group was also observed. Exercise training was efficient in preventing these alterations in the exercise l -NAME group, besides showing a significant hypotensive effect. In conclusion, our results clearly indicated a protector action of moderate intensity exercise on nucleotides and nucleoside hydrolysis and on platelet aggregation, which highlights the exercise training effect to avoid hypertension complications related to ecto-nucleotidase activities. Content Type Journal Article Pages 1-10 DOI 10.1007/s11010-012-1431-7 Authors Andréia Machado Cardoso, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Margarete Dulce Bagatini, Collegiate of Nursing, University of Southern Frontier, Chapecó Campus, Chapecó, SC, Brazil Caroline Curry Martins, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Fátima Hussein Abdalla, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Daniela Zanini, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Roberta Schmatz, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Jessié Gutierres, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Victor Camera Pimentel, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Gustavo Thomé, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Claudio Alberto Martins Leal, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Juliano Marchi Vieira, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Naiara Stefanello, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Fernando da Silva Fiorin, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Jucimara Baldissareli, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Luiz Fernando Freire Royes, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Adriane Bello Klein, Health Basic Sciences Institute, Department of Physiology of Federal University of Rio Grande do Sul, RS, Brazil Vera Maria Morsch, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Maria Rosa Chitolina Schetinger, Post-Graduation Program in Toxicological Biochemistry, Department of Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    The aim of present work was to elucidate the role of actin-depolymerizing factor (ADF), an important regulator of actin cytoskeleton, in the oxidized low-density lipoprotein (ox-LDL)-induced blood–brain barrier (BBB) disruption. The primary mouse brain microvascular endothelial cells (MBMECs) were exposed to ox-LDL. Treatment with LDL served as control. It was found that ADF mRNA level and protein expression were decreased when exposed to ox-LDL in MBMECs. Then, we investigated the influence of ADF overexpression on ox-LDL-treated MBMECs. Structurally, overexpression of ADF inhibited ox-LDL-induced F-actin formation. Functionally, overexpression of ADF attenuated ox-LDL-induced disruption of endothelial barrier marked by restoration of transendothelial electrical resistance, permeability of Evans Blue and expression of tight junction-associated proteins including ZO-1 and occludin, and blocked ox-LDL-induced oxidative stress marked by inhibition of reactive oxygen species (ROS) formation and activity of NADPH oxidase and Nox2 expression. However, overexpression of ADF in control cells had no significant effect on endothelial permeability and ROS formation. In conclusion, overexpression of ADF blocks ox-LDL-induced disruption of endothelial barrier. In addition, siRNA-mediated downregulation of ADF expression aggravated ox-LDL-induced disruption of endothelial barrier and ROS formation. These findings identify ADF as a key signaling molecule in the regulation of BBB integrity and suggest that ADF might be used as a target to modulate diseases accompanied by ox-LDL-induced BBB compromise. Content Type Journal Article Pages 1-8 DOI 10.1007/s11010-012-1415-7 Authors Jun Wang, Department of Neurosurgery, The General Hospital of PLA, 28 Fuxing Road, Beijing, 100853 China Lu Sun, Department of Pathology, The General Hospital of PLA, Beijing, China Yan-Fang Si, Department of Ophthalmology, The 309th Hospital of PLA, Beijing, China Bao-Min Li, Department of Neurosurgery, The General Hospital of PLA, 28 Fuxing Road, Beijing, 100853 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177

Description:    Calcineurin B subunit (CnB) is the regulatory subunit of calcineurin (Cn), a Ca 2+ /calmodulin-dependent serine/threonine protein phosphatase. It has been reported that mice deleting the CnB gene lose nearly all Cn activity and show poor tolerance to cardiac stress; CnB gene expression is downregulated in the hearts of rats that have suffered ischemia/reperfusion (I/R) injury. Therefore, we wonder whether injection of exogenous CnB protein can prevent the rats from suffering I/R injury. In cardiomyocytes, fluorogenic labeling shows that exogenous CnB quickly enters the cell. Pretreatment of cardiomyocytes with CnB reduces apoptosis in response to hypoxia/reoxygenation injury (an in vitro model mimicking ischemia/reperfusion injury), and CsA reverses this effect by inhibiting Cn activity. Furthermore, CnB upregulates Bcl-2 and Bcl-XL expression in the process of hypoxia/reoxygenation injury, which may contribute to protecting cardiomyocytes against apoptosis. In vivo experiments shows that pretreatment with CnB improves cardiac contractile function and reduces the frequency of arrhythmias induced by global I/R injury. These findings reveal a novel function for CnB protein in cardiac stress response and suggest a possible application of CnB in coronary disease therapy. Content Type Journal Article Pages 1-9 DOI 10.1007/s11010-012-1407-7 Authors Junxia Guo, Department of Biochemistry and Molecular Biology, Beijing Key Laboratory of Genetic Engineering Medicine and Biotechnology, Beijing Normal University, Beijing, 100875 China Shengquan Mi, Beijing Key Laboratory of Bioactive Substances and Functional Foods, College of Arts and Science of Beijing Union University, Beijing, 100191 China Jing Li, Department of Biochemistry and Molecular Biology, Beijing Key Laboratory of Genetic Engineering Medicine and Biotechnology, Beijing Normal University, Beijing, 100875 China Wei Liu, Department of Biochemistry and Molecular Biology, Beijing Key Laboratory of Genetic Engineering Medicine and Biotechnology, Beijing Normal University, Beijing, 100875 China Yanxia Yin, Department of Biochemistry and Molecular Biology, Beijing Key Laboratory of Genetic Engineering Medicine and Biotechnology, Beijing Normal University, Beijing, 100875 China Qun Wei, Department of Biochemistry and Molecular Biology, Beijing Key Laboratory of Genetic Engineering Medicine and Biotechnology, Beijing Normal University, Beijing, 100875 China Journal Molecular and Cellular Biochemistry Online ISSN 1573-4919 Print ISSN 0300-8177