Vol. 18 No. 4 December 2013

DOI: 10.2478/s11658-013-0101-4 Volume 18 (2013) pp 479-493
Title ADIPOSE TISSUE-DERIVED STEM CELLS SHOW CONSIDERABLE PROMISE FOR REGENERATIVE MEDICINE APPLICATIONS
Authors Izabela Harasymiak-Krzyżanowska, Alicja Niedojadło, Jolanta Karwat, Lidia Kotuła, Paulina Gil-Kulik, Magdalena Sawiuk and Janusz Kocki*
Abstract The stromal-vascular cell fraction (SVF) of adipose tissue can be an abundant source of both multipotent and pluripotent stem cells, known as adipose-derived stem cells or adipose tissue-derived stromal cells (ADSCs). The SVF also contains vascular cells, targeted progenitor cells, and preadipocytes. Stromal cells isolated from adipose tissue express common surface antigens, show the ability to adhere to plastic, and produce forms that resemble fibroblasts. They are characterized by a high proliferation potential and the ability to differentiate into cells of meso-, ecto- and endodermal origin. Although stem cells obtained from an adult organism have smaller capabilities for differentiation in comparison to embryonic and induced pluripotent stem cells (iPSs), the cost of obtaining them is significantly lower. The 40 years of research that mainly focused on the potential of bone marrow stem cells (BMSCs) revealed a number of negative factors: the painful sampling procedure, frequent complications, and small cell yield. The number of stem cells in adipose tissue is relatively large, and obtaining them is less invasive. Sampling through simple procedures such as liposuction performed under local anesthesia is less painful, ensuring patient comfort. The isolated cellsare easily grown in culture, and they retain their properties over many passages. That is why adipose tissue has recently been treated as an attractive alternative source of stem cells. Essential aspects of ADSC biology and their use in regenerative medicine will be analyzed in this article.
Keywords Adipocyte, Mesenchymal stem cells, Regenerative medicine, Adipose tissue, Stem cell therapy, Adipose-derived stem cells, Stromal cells, Flow cytometry
Address and Contact Information Department of Clinical Genetics, Medical University of Lublin, ul. Radziwiłłowska 11, 20-080 Lublin, Poland
* Author for correspondence. e-mail: janusz.kocki@umlub.pl, tel.: +48 81 5288408, fax: +48 81 5288408

DOI: 10.2478/s11658-013-0102-3 Volume 18 (2013) pp 494-506
Title THE ROLE OF GLYCOGEN SYNTHASE KINASE-3βIN GLIOMA CELL APOPTOSIS INDUCED BY REMIFENTANIL
Authors Jing Xu, Pengjuan Xu, Zhigui Li, Lu Xiao and Zhuo Yang*
Abstract The aim of malignant glioma treatment is to inhibit tumor cell proliferation and induce tumor cell apoptosis. Remifentanil is a clinical anesthetic drug that can activate the N-methyl-D-aspartate (NMDA) receptor. NMDA receptor signaling activates glycogen synthase kinase-3β(GSK-3β). Discovered some 32 years ago, GSK-3βwas only recently considered as a therapeutic target in cancer treatment. The purpose of this study was to assess whether remifentanil can induce the apoptosis of C6 cells through GSK-3β activation. 3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) was used to detect cell viability. Hoechst 33342 staining and flow cytometry were used to detect cell apoptosis. The effect of GSK-3βactivation was detected using a GSK-3βactivation assay kit and 4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione (TDZD-8), a potent and selective small molecule inhibitor of GSK-3β. The MTT assay indicated that remifentanil induced C6 cell death in a concentration- and time-dependent manner. Hoechst 33342 staining and flow cytometry showed that remifentanil significantly induced C6 cell apoptosis. The measurement of GSK-3βactivation showed that remifentanil increased the cellular level of GSK-3β. All of these toxic effects can be attenuated by treatment with TDZD-8. These results suggest that remifentanil is able to induce C6 cell apoptosis through GSK-3βactivation, which provides a basis for its potential use in the treatment of malignant gliomas.
Keywords Remifentanil, Apoptosis, C6 cells, GSK-3β, TDZD-8, MTT, Hoechst 33342, Flow cytometry, NMDA receptor, Glioma
Address and Contact Information College of Medicine, Nankai University, Tianjin 300071, China
* Author for correspondence. e-mail: zhuoyang@nankai.edu.cn, tel.: 86-22-23504364, fax: 86-22-23502554

DOI: 10.2478/s11658-013-0104-1 Volume 18 (2013) pp 507-521
Title In vitro AND in vivo CHARACTERISTICS OF HEPATIC OVAL CELLS MODIFIED WITH HUMAN HEPATOCYTE GROWTH FACTOR
Authors Zhu Li1,§,*, Juan Chen1,§, Li Li2, Jiang-Hua Ran2, Xue-Hua Li1, Zhi-Heng Liu1, Gui-Jie Liu1, Yan-Chao Gao1, Xue-Li Zhang1 and Hiu-Dong Sun1
Abstract Hepatocyte growth factor (HGF) is a multifunctional growth factor that controls cell scattering. It has been suggested that it regulates the proliferation of hepatic oval cells (HOCs). Using a HOC line that stably expresses the human HGF gene (hHGF), we investigated the in vitro proliferation and differentiation characteristics of hHGF-modified HOCs and explored their potential capacity for intrahepatic transplantation. A modified 2-acetylaminofluorene and partial hepatectomy (2-AAF/PH) model was established to activate the proliferation ofoval cells in the rat liver. HOCs were transfected with the pBLAST2-hHGF plasmid and hHGF-carrying HOCs were selected based on blasticidin resistance. The level of hHGF secretion was determined via ELISA. Cell proliferation was determined using the MTT assay. Differentiation was induced by growth factor withdrawal. A two-cuff technique was used for orthotopic liver transplantation, and HOCs or hHGF-modified HOCs were transplanted into the recipients. The levels of biochemical indicators of liver function were measured after transplantation. An HOC line stably expressing hHGF was established. The transfected line showed greater hHGF secretion than normal HOCs. The hHGF gene promoted the proliferation capability of HOCs by reducing the peak time in vitro. The hHGF-modified HOCs differentiated into hepatocytes and bile duct epithelial cells upon growth factor withdrawalin vitro. In addition, hHGF-modified HOC transplantation significantly prolonged the median survival time (MST) and improved the liver function of recipients compared to HOC transplant recipients and nontransplanted controls. Our results indicate that hHGF-modified HOCs may have valuable properties for therapeutic liverregeneration after orthotopic liver transplantation.
Keywords Hepatic oval cells, Hepatocyte growth factor, pBLAST2-hHGF, Plasmid transfection, Proliferation, Differentiation, Liver transplantation, Rats, In vitro, Liver regeneration
Address and Contact Information 1Department of Hepatobiliary Surgery, Liaocheng People’s Hospital, No. 67 Dongchang West Road, Liaocheng 252000, Shandong, China,
2 The First People’s Hospital of Kunming, No. 504 Qinnian Road, Kunming 650011, Yunnan, China
§ Zhu Li and Juan Chen contributed equally to this study
* Author for correspondence. e-mail: lllllzhu@126.com, tel.: 0086-0635-8272383, fax: 0086-0635-8272246

DOI: 10.2478/s11658-013-0103-2 Volume 18 (2013) pp 522-537
Title PROTEOLYTIC ACTIVATION OF Chlamydia trachomatis HTRA IS MEDIATED BY PDZ1 DOMAIN INTERACTIONS WITH PROTEASE DOMAIN LOOPS L3 AND LC AND BETA STRAND β5
Authors James W. Marsh1, William B. Lott1, Joel D.A. Tyndall2* and Wilhelmina M. Huston1*
Abstract Chlamydia trachomatisis a bacterial pathogen responsible for one of the most prevalent sexually transmitted infections worldwide. Its unique development cycle has limited our understanding of its pathogenic mechanisms. However, CtHtrA has recently been identified as a potential C. trachomatis virulence factor. CtHtrA is a tightly regulated quality control protein with a monomeric structural unit comprised of a chymotrypsin-like protease domain and two PDZ domains. Activation of proteolytic activity relies on the C-terminus of the substrate allosterically binding to the PDZ1 domain, which triggers subsequent conformational change and oligomerization of the protein into 24-mers enabling proteolysis. This activation is mediated by a cascade of precise structural arrangements, but the specific CtHtrA residues and structural elements required to facilitate activation are unknown. Using in vitro analysis guided by homology modeling, we show that the mutation of residues Arg362 and Arg224, predicted to disrupt the interaction between the CtHtrA PDZ1 domain and loop L3, and between loop L3 and loop LD, respectively, are critical for the activation of proteolytic activity. We also demonstrate that mutation to residues Arg299 and Lys160, predicted to disrupt PDZ1 domain interactions with protease loop LC and strand β5, are also able to influence proteolysis, implying their involvement in the CtHtrA mechanism of activation. This is the first investigation of protease loop LC and strand β5 with respect to their potential interactions with the PDZ1domain. Given their high level of conservation in bacterial HtrA, these structural elements may be equally significant in the activation mechanism of DegP and other HtrA family members.
Keywords Chlamydia, HtrA, DegP, Protease, Oligomerization
Address and Contact Information 1 Institute of Health and Biomedical Innovation (IHBI), Queensland University of Technology (QUT), 60 Musk Avenue, Kelvin Grove, Queensland, Australia, 4059,
2 National School of Pharmacy, University of Otago, PO Box 56, Dunedin, New Zealand, 9054
* Authors for correspondence. Wilhelmina (Willa) Huston – e-mail: w.huston@qut.edu.au, tel.: +61 7 3138 6258, fax: +61 7 3138 6030, Joel Tyndall – e-mail: joel.tyndall@otago.ac.nz, tel.: +64 3 4797293, fax: +64 3 4797034

DOI: 10.2478/s11658-013-0105-0 Volume 18 (2013) pp 538-554
Title TELOMERASE AND ITS EXTRACURRICULAR ACTIVITIES
Authors Rishi Kumar Jaiswal, Pramod Kumar and Pramod Kumar Yadava*
Abstract The classical activity of telomerase is to synthesize telomeric repeats and thus maintain telomere length, which in turn ensures chromosome stability and cellular proliferation. However, there is growing evidence that implicates telomerase in many other functions that are independent of TERC being used as its template. Telomerase has an RNA-dependent RNA polymerase (RdRP) activity in the mitochondria. Other than viral RdRPs, it is the only RNAdependent RNA polymerase that has been identified in mammals. It also plays a role in the Wnt signaling pathway by acting as a transcriptional modulator. Telomerase acts as a reverse transcriptase independent of its core subunit, TERC. Studies indicate that telomerase is also involved in apoptosis and DNA repair.
Keywords Telomerase, Telomere, RNA-dependent RNA polymerase, TERT, TERC, Apoptosis
Address and Contact Information Applied Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi 110067, India
* Author for correspondence. e-mail: pky0200@mail.jnu.ac.in, pkyadava1953@gmail.com, tel.: +91-11-26704520, fax: +91-11-26742558

DOI: 10.2478/s11658-013-0106-z Volume 18 (2013) pp 555-578
Title REGULATION OF THE UNFOLDED PROTEIN RESPONSE BY microRNAs
Authors Sylwia Bartoszewska1,§, Kinga Kochan2,§, Piotr Madanecki2, Arkadiusz Piotrowski2, Renata Ochocka2, James F. Collawn3 and Rafal Bartoszewski2,*
Abstract The unfolded protein response (UPR) is an adaptive response to the stress that is caused by an accumulation of misfolded proteins in the lumen of the endoplasmic reticulum (ER). It is an important component of cellular homeostasis. During ER stress, the UPR increases the protein-folding capacity of the endoplasmic reticulum to relieve the stress. Failure to recover leads to apoptosis. Specific cellular mechanisms are required for the cellular recovery phase after UPR activation. Using bioinformatics tools, we identified a number of microRNAs that are predicted to decrease the mRNA expression levels for a number of critical components of the UPR. In this review, we discuss the potential role of microRNAs as key regulators of this pathway and describe how microRNAs may play an essential role in turning off the UPR after the stress has subsided.
Keywords MicroRNA, Unfolded protein response, Adaptive response, Endoplasmic reticulum stress
Address and Contact Information 1 Department of Inorganic Chemistry, Medical University of Gdansk, Poland,
2 Department of Biology and Pharmaceutical Botany, Medical University of Gdansk, Poland,
3 Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham, USA
§ These authors contributed equally to this work.
* Author for correspondence. Department of Biology and Pharmaceutical Botany, Medical University of Gdansk, Hallera 107, 80-416 Gdansk, Poland, e-mail: rafalbar@gumed.edu.pl, tel.: + 48 58 349 32 14, fax: + 48 58 349 32 11

DOI: 10.2478/s11658-013-0108-x Volume 18 (2013) pp 579-594
Title MEMBRANE POTENTIAL-DEPENDENT BINDING OF POLYSIALIC ACID TO LIPID MONOLAYERS AND BILAYERS
Authors Krzysztof Nowotarski, Karolina Sapoń, Monika Kowalska, Tadeusz Janas and Teresa Janas*
Abstract Polysialic acids are linear polysaccharides composed of sialic acid monomers. These polyanionic chains are usually membrane-bound, and are expressed on the surfaces of neural, tumor and neuroinvasive bacterial cells. We used toluidine blue spectroscopy, the Langmuir monolayer technique and fluorescence spectroscopy to study the effects of membrane surface potential and transmembrane potential on the binding of polysialic acids to lipid bilayers and monolayers. Polysialic acid free in solution was added to the bathing solution to assess the metachromatic shift inthe absorption spectra of toluidine blue, the temperature dependence of the fluorescence anisotropy of DPH in liposomes, the limiting molecular area in lipid monolayers, and the fluorescence spectroscopy of oxonol V in liposomes. Our results show that both a positive surface potential and a positive transmembrane potential inside the vesicles can facilitate the binding of polysialic acid chains to model lipid membranes. These observations suggest that these membrane potentials can also affect the polysialic acid-mediated interaction between cells.
Keywords Polysialic acid, Polyanion, Lipid bilayer, Lipid monolayer, Membrane potential, Liposome, DPH anisotropy
Address and Contact Information Department of Biotechnology and Molecular Biology, University of Opole, Kominka 6, 45-032 Opole, Poland
* Author for correspondence. e-mail: tjanas@uni.opole.pl, tel.: 48-77-4016045, fax: 48-77-4016051

DOI: 10.2478/s11658-013-0109-9 Volume 18 (2013) pp 595-611
Title THE CLINICOPATHOLOGICAL SIGNIFICANCE OF LAMIN A/C, LAMIN B1 AND LAMIN B RECEPTOR mRNA EXPRESSION IN HUMAN BREAST CANCER
Authors Umar Wazir1,2, Mai Hassan Ahmed3,4, Joanna M. Bridger3, Amanda Harvey4, Wen G. Jiang5, Anup K. Sharma1 and Kefah Mokbel1,2,*
Abstract Lamin A/C (LMNA), lamin B1 (LMNB1) and lamin B receptor (LBR) have key roles in nuclear structural integrity and chromosomal stability. In this study, we have studied the relationships between the mRNA expressions of A-type lamins, LMNB1 and LBR and the clinicopathological parameters in human breast cancer. Samples of breast cancer tissues (n = 115) and associated non-cancerous tissue (ANCT; n = 30) wereassessed using reverse transcription and quantitative PCR. Transcript levels were correlated with clinicopathological data. Higher levels of A-type laminsand LMNB1 mRNA expression were seen in ANCT. Higher lamin A/C expression was associated with the early clinical stage (TNM1 vs. TNM3 – 13 vs. 0.21; p = 0.0515), with better clinical outcomes (disease-free survival vs. mortality – 11 vs. 1; p = 0.0326), and with better overall (p = 0.004) and disease-free survival (p = 0.062). The expression of LMNB1 declined with worsening clinicaloutcome (disease-free vs. mortalities – 0.0011 vs. 0.000; p = 0.0177). LBR mRNA expression was directly associated with tumor grade (grade 1 vs. grade 3 – 0.00 vs. 0.00; p = 0.0479) and Nottingham Prognostic Index (NPI1 vs. NPI3– 0.00 vs. 0.00; p = 0.0551). To the best of our knowledge, this is the first study to suggest such a role for A-type lamins, lamin B1 and LBR in human breastcancer, identifying an important area for further research.
Keywords Lamin A/C, Lamin B, Lamin B receptor, Breast cancer, qPCR, Chromosomal instability,Cell senescence, Cell cycle, DNA repair, Ageing
Address and Contact Information 1 The London Breast Institute, Princess Grace Hospital, London, UK,
2 Department of Breast Surgery, St. George’s Hospital and Medical School, University of London, London, UK,
3 Centre for Cell & Chromosome Biology, 4 Brunel Institute for Cancer Genetics and Pharmacogenomics, School of Health Sciences and Social Care, Brunel University, Uxbridge, London, UK,
5 Metastasis and Angiogenesis Research Group, University Department of Surgery, Cardiff University School of Medicine, Cardiff University, Cardiff, Wales, UK
* Author for correspondence. London Breast Institute, the Princess Grace Hospital, 45 Nottingham Place, London W1U 5NY, UK, e-mail: kefahmokbel@hotmail.com

DOI: 10.2478/s11658-013-0107- Volume 18 (2013) pp 612-630
Title THE EXPRESSION OF THE EOTAXINS IL-6 AND CXCL8 IN HUMAN EPITHELIAL CELLS FROM VARIOUS LEVELS OF THE RESPIRATORY TRACT
Authors Magdalena Paplińska-Goryca*, Patrycja Nejman-Gryz, Ryszarda Chazan and Hanna Grubek-Jaworska
Abstract Airway epithelium acts as multifunctional site of response in the respiratory tract. Epithelial activity plays an important part in the pathophysiology of obstructive lung disease. In this study, we compare normal human epithelial cells from various levels of the respiratory tract in terms of their reactivity to pro-allergic and pro-inflammatory stimulation. Normal human nasal, bronchial and small airway epithelial cells were stimulated with IL-4 and IL-13. The expressions of the eotaxins IL-6 and CXCL8 were evaluated at the mRNA and protein levels. The effects of pre-treatment with IFN-γ on the cell reactivity were measured, and the responses to TNF-α, LPS and IFN-γ were evaluated. All of the studied primary cells expressed CCL26, IL-6 and IL-8 after IL-4 or IL-13 stimulation. IFN-γ pre-treatment resulted in decreased CCL26 and increased IL-6 expression in the nasal and small airway cells, but this effect was not observed in the bronchial cells. IL-6 and CXCL8 were produced in varying degrees by all of the epithelial primary cells in cultures stimulated with TNF-α, LPS or IFN-γ. We showed that epithelial cells from the various levels of the respiratory tract act in a united way, responding in a similar manner to stimulation with IL-4 and IL-13, showing similar reactivity to TNF-α and LPS, and giving an almost unified response to IFN-γ pre-stimulation.
Keywords CXCL8, Eotaxin-3, Inflammation, Interferon gamma, Interleukin 6, Respiratory epithelium, United airways
Address and Contact Information Medical University of Warsaw, Department of Internal Medicine, Pneumonology and Allergology, Banacha 1a, 02-097 Warsaw, Poland
* Author for correspondence. e-mail: mpaplinska@wum.edu.pl, tel.: +48225991241, fax: +48225991560.

DOI: 10.2478/s11658-013-0110-3 Volume 18 (2013) pp 631-638
Title DETECTION OF A NOVEL MUTATION IN EXON 20 OF THE BRCA1 GENE
Authors Abhijit Chakraborty1, Atul Katarkar2, Keya Chaudhuri2, Ashis Mukhopadhyay3 and Jayasri Basak1,*
Abstract Hereditary breast cancer constitutes 5-10% of all breast cancer cases. Inherited mutations in the BRCA1 and BRCA2 tumor-suppressor genes account for the majority of hereditary breast cancer cases. The BRCA1 C-terminal region (BRCT) has a functional duplicated globular domain, which helps with DNA damage repair and cell cycle checkpoint protein control. More than 100 distinct BRCA1 missense variants with structural and functional effects have been documented within the BRCT domain. Interpreting the results of mutation screening of tumor-suppressor genes that can have high-risk susceptibility mutations is increasingly important in clinical practice. This study includes a novel mutation, p.His1746 Pro (c.5237A>C), which was found in BRCA1 exon 20 of a breast cancer patient. In silico analysis suggests that this mutation could alter the stability and orientation of the BRCT domain and the differential binding of the BACH1 substrate.
Keywords Breast cancer, BRCA1, BRCT repeat, BACH1, Cancer predisposition, DNA sequencing, Docking, Exon 20, In silico analysis, Missense mutation
Address and Contact Information 1 Department of Molecular Biology, Netaji Subhas Chandra Bose Cancer Research Institute (NCRI), Kolkata, India,
2 Molecular & Human Genetics Division, Indian Institute of Chemical Biology (IICB), Kolkata, India,
3 Department of Oncology, Netaji Subhas Chandra Bose Cancer Research Institute (NCRI), Kolkata, India
* Author for correspondence. E-mail: hmcwt@dataone.in; ncri.molecularbiology@gmail.com; abhijit.drems@gmail.comtel.: +91-33-2229 1049/5628, fax: +91-33-2226 4704

DOI: 10.2478/s11658-013-0116-x Volume 18 (2013) pp 639
Erratum to the article GABA EXISTS AS A NEGATIVE REGULATOR OF CELL PROLIFERATION IN SPERMAOGONIAL STEM CELLS

published in:

Cellular & Molecular BiologyLetters, Vol. 18. No. 2, 2013, pp. 149-162. DOI: 10.2478/s11658-013-0081-4

The original version of the article title unfortunately contained mistake. The title should be:

GABA EXISTS AS A NEGATIVE REGULATOR OF CELL PROLIFERATION IN SPERMATOGONIAL STEM CELLS