Vol. 16 No. 2 June 2011

DOI: 10.2478/s11658-011-0001-4 Volume 16 (2011) pp 201-213
Authors Xiangjun Huang§, Hongwu Luo§*, Fei Zhou Huang and Xun Yang Liu
Abstract Human amniotic epithelial cells (hAECs) are a recently identified type of stem cell. Thanks to their ready availability and the lower risk of teratoma formation, hAECs have been studied and tested for a variety of human disease treatments and tissue reconstruction efforts. This aim of this study was to establish a stable tracking system to further monitor hAECs in vivo after transplantation. hAECs were isolated from the placentas of patients who visited the Hunan Province Maternity and Child Care Hospitals between Jan 2008 and Jan 2009. Using the classic transfection/infection technique, we successfully introduced green fluorescent protein (GFP) into cultured hAECs with an adeno- associated virus (AAV) vector. The initial preparation of the AAV-GFP virus stock was titrated using HT1081 cells, and further used for the infection of hAECs. GFP+ hAECs preserve the capacity of differentiation into hepatocyte- like cells with the expression of cytokeratin-18 (CK18) and albumin (ALB). AAV-GFP virus-infected hAECs were transplanted through the spleen into severe combined immune deficiency (SCID) mice via hepatectomy. Four weeks later, the GFP and human albumin expressions were examined in multiple organs through immunoflourence staining. In culture, over 50% of the hAECs were GFP-positive 3 days after infection. Following transplantation, AAV-GFP- infected hAECs survived and continued to express GFP in the host for up to 4 weeks. These cells were primarily found in the spleen and liver, expressing human albumin. This study provides a feasible and stable system to track hAECs. It may prove useful to further identify their biological characteristics after transplantation and to elucidate their beneficial roles for therapeutic purposes.
Keywords Human amniotic epithelial cells, Adeno-associated virus, Green fluorescent protein, Transplantation, Tracking system
Address and Contact Information Department of Hepatobiliary Surgery, Xiangya Third Hospital, Central South University, No. 138, Hexi Tongzipo Road, Changsha Hunan 41300, P. R. China
§ These Authors equally contributed to this paper
* Author for correspondence. hxiangj168@126.com, phone: 0731-8861-8230, fax: 0731- 8861-8030
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DOI: 10.2478/s11658-011-0003-2 Volume 16 (2011) pp 214-225
Authors Alexander Spassov1,*, Tomasz Gredes1, Tomasz Gedrange1, Silke Lucke1, Dragan Pavlovic2 and Christiane Kunert-Keil1
Abstract The activities of myogenic regulatory factors (MRF) and muscle growth factors increase in muscle that is undergoing regeneration, and may correspond to some specific changes. Little is known about the role of MRFs in masticatory muscles in mdx mice (the model of Duchenne muscular dystrophy) and particularly about their mRNA expression during the process of muscle regeneration. Using Taqman RT-PCR, we examined the mRNA expression of the MRFs myogenin and MyoD1 (myogenic differentiation 1), and of the muscle growth factors myostatin, IGF1 (insulin-like growth factor) and MGF (mechano- growth factor) in the masseter, temporal and tongue masticatory muscles of mdx mice (n = 6 to 10 per group). The myogenin mRNA expression in the mdx masseter and temporal muscle was found to have increased (P < 0.05), whereas the myostatin mRNA expressions in the mdx masseter (P < 0.005) and tongue (P < 0.05) were found to have diminished compared to those for the controls. The IGF and MGF mRNA amounts in the mdx mice remained unchanged. Inside the mdx animal group, gender-related differences in the mRNA expressions were also found. A higher mRNA expression of myogenin and MyoD1 in the mdx massterer and temporal muscles was found in females in comparison to males, and the level of myostatin was higher in the masseter and tongue muscle (P < 0.001 for all comparisons). Similar gender-related differences were also found within the control groups. This study reveals the intermuscular differences in the mRNA expression pattern of myogenin and myostatin in mdx mice. The existence of these differences implies that dystrophinopathy affects the skeletal muscles differentially. The finding of gender-related differences in the mRNA expression of the examined factors may indicate the importance of hormonal influences on muscle regeneration. 
Keywords Mdx mice, Dystrophy, Masticatory muscles, MyoD1, Myogenin
Address and Contact Information 1Department of Orthodontics, Faculty of Medicine, University of Greifswald, Rotgerber Str. 8, 17475 Greifswald, Germany,
2Department of Anaesthesiology and Intensive Care Medicine, Faculty of Medicine, University of Greifswald, Friedrich-Loeffler Str. 23c, 17475 Greifswald, Germany
* Author for correspondence: e-mail: alexspas@uni-greifswald.de; phone: +49 3834 867119; fax: +49 3834 867113
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DOI: 10.2478/s11658-011-0002-3 Volume 16 (2011) pp 226-235
Authors Yanping Fu1,§ , Gang Shi2,§, Yong Wu3, Yasuyuki Kawai4, Qing Tian1, Linlin Yue1, Qinjie Xia1, Isamu Miyamori4 and Chunyuan Fan1,5*
Abstract High aldosterone (Ald) levels can induce hypertrophy of vascular smooth muscle cells (VSMCs), which carries high risks of heart failure. A previous study showed that Ald induces hypertrophy of VSMCs by up- regulating NOX1, a catalytic subunit of NADPH oxidase that produces superoxides. However, the precise mechanism remains unknown. Diphenylene iodonium (DPI) is known as an inhibitor of complex I in the mitochondrial respiratory chain, and it was also found to almost completely suppress the induction of NOX1 mRNA and the phosphorylation of activating transcription factor (ATF-1) by PGF2α or PDGF in a rat VSMC cell line. In this study, we found that the Ald-induced phosphorylation of ATF-1 and NOX1 expression was significantly suppressed by DPI. Silencing of ATF-1 gene expression attenuated the induction of NOX1 mRNA expression, and over-expression of ATF-1 restored Ald-induced NOX1 expression. On the basis of this data, we show that the mitochondria mediate aldosterone-induced NOX1 gene expression in an ATF-1-dependent manner.
Keywords Aldosterone, Mitochondria, ATF-1, NOX1, VSMC
Address and Contact Information 1Department of Nephrology, West China Hospital of Sichuan University, Guoxue Lane 37, WuHou Zone, Chengdu, 610041, China,
2State Key Laboratory of Biotherapy, SiChuan University, 1 Keyuan Road 4, GaoPeng Street, High Technological Development Zone, Chengdu, 610041, China,
3The Second Out-Patient Institutions of the Chengdu Military Region, Chengdu 610041, China,
4Third Department of Internal Medicine, Faculty of Medical Sciences, University of Fukui, Fukui, Japan,
5The Second People's Hospital in Chengdu, Chengdu 610041, Chin
§These authors contributed equally to this work
* Author for correspondence. e-mail: fanchunyuan2009@yahoo.cn, phone: +86-28-81812896, fax: +86-28-85164005
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DOI: 10.2478/s11658-011-0005-0 Volume 16 (2011) pp 236-257
Authors Malgorzata Witkowska-Zimny* and Katarzyna Walenko
Abstract This is a review of the growing scientific interest in the developmental plasticity and therapeutic potential of stromal cells isolated from adipose tissue. Adipose-derived stem/stromal cells (ASCs) are multipotent somatic stem cells that are abundant in fat tissue. It has been shown that ASCs can differentiate into several lineages, including adipose cells, chondrocytes, osteoblasts, neuronal cells, endothelial cells, and cardiomyocytes. At the same time, adipose tissue can be harvested by a minimally invasive procedure, which makes it a promising source of adult stem cells. Therefore, it is believed that ASCs may become an alternative to the currently available adult stem cells (e.g. bone marrow stromal cells) for potential use in regenerative medicine. In this review, we present the basic information about the field of adipose-derived stem cells and their potential use in various applications.
Keywords Adult stem cells, Adipose-derived stem cells/stromal cells, Adipose tissue, Regenerative medicine
Address and Contact Information Department of Biophysics and Human Physiology, Medical University of Warsaw, Chalubinskiego 5, 02-004 Warsaw, Poland
* Author for correspondence. e-mail: mwitkowska@wum.edu.pl, tel. +48 22 628 63 34, fax: +48 22 628 78 46
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DOI: 10.2478/s11658-011-0004-1 Volume 16 (2011) pp 258-263
Authors Dariusz Plewczynski1* and Tomas Klingstrom1,2
Abstract Studying the interactome is one of the exciting frontiers of proteomics, as shown lately at the recent bioinformatics conferences (for example ISMB 2010, or ECCB 2010). Distribution of data is facilitated by a large number of databases. Metamining databases have been created in order to allow researchers access to several databases in one search, but there are serious difficulties for end users to evaluate the metamining effort. Therefore we suggest a new standard, “Good Interaction Data Metamining Practice” (GIDMP), which could be easily automated and requires only very minor inclusion of statistical data on each database homepage. Widespread adoption of the GIDMP standard would provide users with:
  • a standardized way to evaluate the statistics provided by each metamining database, thus enhancing the end-user experience;
  • a stable contact point for each database, allowing the smooth transition of statistics;
  • a fully automated system, enhancing time- and cost-effectiveness.
The proposed information can be presented as a few hidden lines of text on the source database www page, and a constantly updated table for a metamining database included in the source/credits web page.
Keywords Proteins, Interactome, Pathways, Signaling, Metamining, Literature curation, Protein-protein interaction, Bioinformatics, Systems biology
Address and Contact Information 1Interdisciplinary Centre for Mathematical and Computational Modelling, University of Warsaw, ul. Pawinskiego 5a, 02-106 Warsaw, Poland,
2Master of Science Programme in Molecular Biotechnology Engineering at Uppsala University, Norbyvägen 14, 752 36 Uppsala, Sweden
* Author for correspondence. e-mail: darman@icm.edu.pl
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DOI: 10.2478/s11658-011-0008-x Volume 16 (2011) pp 264-278
Authors Piyali Chatterjee1, Subhadip Basu2, Mahantapas Kundu2, Mita Nasipuri2 and Dariusz Plewczynski3*
Abstract Protein-protein interactions (PPI) control most of the biological processes in a living cell. In order to fully understand protein functions, a knowledge of protein-protein interactions is necessary. Prediction of PPI is challenging, especially when the three-dimensional structure of interacting partners is not known. Recently, a novel prediction method was proposed by exploiting physical interactions of constituent domains. We propose here a novel knowledge-based prediction method, namely PPI_SVM, which predicts interactions between two protein sequences by exploiting their domain information. We trained a two-class support vector machine on the benchmarking set of pairs of interacting proteins extracted from the Database of Interacting Proteins (DIP). The method considers all possible combinations of constituent domains between two protein sequences, unlike most of the existing approaches. Moreover, it deals with both single-domain proteins and multi-domain proteins; therefore it can be applied to the whole proteome in high- throughput studies. Our machine learning classifier, following a brainstorming approach, achieves accuracy of 86%, with specificity of 95%, and sensitivity of 75%, which are better results than most previous methods that sacrifice recall values in order to boost the overall precision. Our method has on average better sensitivity combined with good selectivity on the benchmarking dataset. The PPI_SVM source code, train/test datasets and supplementary files are available freely in the public domain at: http://code.google.com/p/cmater-bioinfo/.
Keywords Protein-protein interaction, Domain-frequency values, Domain- domain interaction affinity value, Proteome, Interactome, Brainstorming, Machine learning, Consensus, DIP, Protein domains, Sequences, Structures, Protein-protein complexes
Address and Contact Information 1Department of Computer Science and Engineering, Netaji Subhash Engineering College, Garia, Kolkata - 700152, India,
2Department of Computer Science and Engineering, Jadavpur University, Kolkata - 700032, India,
3Interdisciplinary Centre for Mathematical and Computational Modelling, University of Warsaw, ul. Pawinskiego 5a, 02-106 Warsaw, Poland
* Author for correspondence. e-mail: darman@icm.edu.pl
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DOI: 10.2478/s11658-011-0006-z Volume 16 (2011) pp 279-295
Authors Xuejiao Zhang1,3, Cheng Li1, Huiling Gao1, Hiroaki Nabeka1, Tetsuya Shimokawa1, Hiroyuki Wakisaka1, Seiji Matsuda1 and Naoto Kobayashi2*
Abstract We investigated the effects of Rho-associated kinase (ROCK) on migration and cytoskeletal organization in primary human osteoblasts and Saos- 2 human osteosarcoma cells. Both cell types were exposed to two different ROCK inhibitors, Y-27632 and HA-1077. In the improved motility assay used in the present study, Y-27632 and HA-1077 significantly increased the migration of both osteoblasts and osteosarcoma cells on plastic in a dose-dependent and reversible manner. Fluorescent images showed that cells of both types cultured with Y-27632 or HA-1077 exhibited a stellate appearance, with poor assembly of stress fibers and focal contacts. Western blotting showed that ROCK inhibitors reduced myosin light chain (MLC) phosphorylation within 5 min without affecting overall myosin light-chain protein levels. Inhibition of ROCK activity is thought to enhance the migration of human osteoblasts through reorganization of the actin cytoskeleton and regulation of myosin activity. ROCK inhibitors may be potentially useful as anabolic agents to enhance the biocompatibility of bone and joint prostheses.
Keywords Osteoblast, Migration, ROCK inhibitor, Cytoskeleton, Stress fiber, Focal contact, MLC phosphorylation
Address and Contact Information 1Department of Anatomy and Embryology, Ehime University Graduate School of Medicine,
2Medical Education Center, Ehime University School of Medicine,Shitsukawa, To-on City, Ehime 791-0295, Japan,
3Current address: Shenyang Women and Children Health Center, Shenyang city, Liaoning, 110032, China
* Author for correspondence. e-mail: naoto@m.ehime-u.ac.jp, phone: 81-89-960-5928, fax: 81-89-960-5931
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DOI: 10.2478/s11658-011-0009-9 Volume 16 (2011) pp 296-327
Authors Marta Nekulova1, Jitka Holcakova1, Philip Coates2 and Borivoj Vojtesek1*
Abstract The transcription factor p63 has important functions in tumorigenesis, epidermal differentiation and stem cell self-renewal. The TP63 gene encodes multiple protein isoforms that have different or even antagonistic roles in these processes. The balance of p63 isoforms, together with the presence or absence of the other p53 family members, p73 and p53, has a striking biological impact. There is increasing evidence that interactions between p53-family members, whether cooperative or antagonistic, are involved in various cell processes. This review summarizes the current understanding of the role of p63 in tumorigenesis, metastasis, cell migration and senescence. In particular, recent data indicate important roles in adult stem cell and cancer stem cell regulation and in the response of cancer cells to therapy.
Keywords p63, TAp63, ΔNp63, p53 family, Cancer, Stem cells
Address and Contact Information 1Masaryk Memorial Cancer Institute, Zluty kopec 7, 65653 Brno, Czech Republic,
2Division of Medical Sciences, University of Dundee, DD1 9SY, UK
* Author for correspondence. e-mail: vojtesek@mou.cz
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DOI: 10.2478/s11658-011-0007-y Volume 16 (2011) pp 328-341
Authors Blaž Banič1,§, Damijan Nipič1,§, Dušan Šuput1 and Irina Milisav1, 2*
Abstract We demonstrate here that distribution of caspase-9 influences the pathway of apoptosis triggering, since caspase-9 is activated efficiently only when it is distributed solely in the cytosol. Caspase-9 moves to the nuclei in a response to cell stress during isolation of primary hepatocytes; this is called preapoptotic cell stress response. The dimethyl sulfoxide (DMSO) treatment cannot prevent the migration of caspase-9 into the nuclei when it is added to primary hepatocytes immediately after isolation; however, it can trigger redistribution of caspase-9 from the nuclei into the cytosol when added 1 day post-isolation. This redistribution is temporary, since caspase-9 returns to the nuclei within 48 hours of DMSO treatment. Thereafter, some caspase-9 is retained in the nuclei of DMSO-treated hepatocytes for longer than in the nuclei of untreated hepatocytes. By measuring caspase activities, we demonstrate that the addition of DMSO to cell culture medium can temporarily normalize the susceptibility of hepatocytes for apoptosis triggering through the intrinsic pathway. DMSO contributes also to the prolonged pathway inactivation, i.e., by extending preapoptotic cell stress response. We propose that DMSO extends the survival of primary hepatocytes by modulating preapoptotic cell stress response, which could be exploited for extending the lifespan of other primary cell cultures.
Keywords Apoptosis, Preapoptotic cell stress response, DMSO, Hepatocytes, Caspase-9, Intrinsic pathway
Address and Contact Information 1Institute of Pathophysiology, Faculty of Medicine, University of Ljubljana, Zaloška 4, SI-1000 Ljubljana, Slovenia,
2Faculty of Health Sciences, University of Ljubljana, Slovenia
§ These authors contributed equally to this work
* Author for correspondence. e-mail: irina.milisav@mf.uni-lj.si, phone: +386-1-543-7089, fax +386-1-543-7021
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DOI: 10.2478/s11658-011-0010-3 Volume 16 (2011) pp 342-358
Authors Teresa Janas1,2* and Tadeusz Janas2
Abstract Noncovalent DIDS binding to Band 3 (AE1) protein in human erythrocyte membranes, modified by non-penetrating, water soluble 1-ethyl-3- (4-azonia-4,4-dimethylpentyl)-carbodiimide iodide (EAC), was studied at 0o C in the presence of 165 mM KCl. Under experimental conditions applied up to (48 ± 5) % of irreversible chloride self-exchange inhibition was observed. The apparent dissociation constant, KD, for “DIDS-Band 3” complex, determined from the chloride transport experiments, was (34 ± 3) nM and (80 ± 12) nM for control and EAC-treated resealed ghosts, respectively. The inhibition constant, Ki, for DIDS was (35 ± 6) nM and (60 ± 8) nM in control and EAC-treated ghosts, respectively. The reduced affinity for DIDS reversible binding was not a result of negative cooperativity of DIDS binding sites of Band 3 oligomer since Hill’s coefficients were indistinguishable from 1 (within the limit error) both for control and EAC-treated ghosts. By using tritium-labeled DIDS, 4,4’-diisothiocyanato-2,2’-stilbenedisulfonate ([3H]DIDS), the association rate constant, k+1 (M-1s-1), was measured. The mean values of (4.3 ± 0.7) x 105 M-1s-1 for control and (2.7 ± 0.7) x 105 M-1s-1 for EAC-treated ghosts were obtained. The mean values for KD, evaluated from [3H]DIDS binding measurements, were (37 ± 9) nM and (90 ± 21) nM for control and EAC-modified ghosts, respectively. The results demonstrate that EAC modification of AE1 reduces about 2-fold the affinity of AE1 for DIDS. It is suggested that half of the subunits are modified near the transport site by EAC.
Keywords Band 3, Carbodiimide, Dissociation constant, Erythrocyte membrane, Stilbenedisulfonate
Address and Contact Information 1Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, Colorado, USA,
2Department of Biotechnology and Molecular Biology, University of Opole, Opole, Poland
* Author for correspondence. e-mail: teresa.janas@colorado.edu, phone: 303-492-8377, fax: 303-492-7744
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