Vol. 13 No. 2 June 2008

DOI: 10.2478/s11658-007-0046-6 Volume 13 (2008) pp 155-181
Authors Mariusz Piechota1, Maciej Banach2*, Anna Jacoń3 and Jacek Rysz4
Abstract The natriuretic peptide family comprises atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), C-type natriuretic peptide (CNP), dendroaspis natriuretic peptide (DNP), and urodilatin. The activities of natriuretic peptides and endothelins are strictly associated with each other. ANP and BNP inhibit endothelin-1 (ET-1) production. ET-1 stimulates natriuretic peptide synthesis. All natriuretic peptides are synthesized from polypeptide precursors. Changes in natriuretic peptides and endothelin release were observed in many cardiovascular diseases: e.g. chronic heart failure, left ventricular dysfunction and coronary artery disease.
Keywords Atrial natriuretic peptide, Brain natriuretic peptide, C-type natriuretic peptide, Dendroaspis natriuretic peptide, Urodilatin, Endothelin-1
Address and Contact Information 1Department of Anaesthesiology and Intensive Care Unit, Boleslaw Szarecki University Hospital No. 5 in Łódź, Medical University in Łódź, Poland,
2Department Cardiology, 1st Chair of Cardiology and Cardiac Surgery, University Hospital No. 3 in Łódź, Medical University in Łódź, Poland,
3Department of Health Protection Policy, Medical University of Łódź, Poland
42nd Department of Family Medicine, University Hospital No. 2 in Łódź, Medical University in Łódź, Poland
* Author for correspondence: Department of Cardiology, Medical University of Łódź, Sterlinga 1/3, 91-425 Łódź, Poland, tel/fax: +48 42 6364 471, e-mail: m.banach@termedia.pl
[Rozmiar: 1332 bajtów]

DOI: 10.2478/s11658-007-0045-7 Volume 13 (2008) pp 182-194
Authors Sylwia Dragojew1, Agnieszka Marczak1, Janusz Maszewski2, Krzysztof Ilnicki3 and Zofia Jóźwiak1*
Abstract In this study, we investigated apoptosis induced in human trisomic and diabetic fibroblasts by daunorubicin (DNR) and its derivative, idarubicin (IDA). The cells were incubated with DNR or IDA for 2 h and then cultured in a drug-free medium for a further 2-48 h. The apoptosis in the cultured cell lines was assessed by biochemical analysis. We found that both drugs induced a timedependent loss of mitochondrial membrane potential, and a significant increase in intracellular calcium and caspase-3 activity. Mitochondrial polarization and changes in the level of intracellular calcium were observed during the first 2-6 h after drug treatment. Caspase-3 activation occurred in the late stages of the apoptotic pathway. Our findings also demonstrated that idarubicin was more cytotoxic and more effective than daunorubicin in inducing apoptosis in trisomic and diabetic fibroblasts.
Keywords Daunorubicin, Idarubicin, Apoptosis, Fibroblasts, Down’s syndrome, Diabetes
Address and Contact Information 1Department of Thermobiology, University of Łódź, Banacha 12/16, 90-237 Łódź, Poland,
2Department of Cytophysiology, University of Łódź, Pilarskiego 14/16, 90-237 Łódź, Poland,
3Department of Medical Genetics, Institute of the Centrum of Child Health, Al. Dzieci Polskich 20, 04-730 Warszawa, Poland
*Author for correspondence; e-mail: zjozwiak@biol.uni.lodz.pl
[Rozmiar: 1332 bajtów]

DOI: 10.2478/s11658-007-0044-8 Volume 13 (2008) pp 195-211
Authors Kelly Schweitzer1, Erran Li2, Venkataramana Sidhaye1, Virginia Leitch1, Sergey Kuznetsov1 and Landon S. King1*
Abstract Altered tissue water homeostasis may contribute to edema formation during various stresses including bacterial infection. We observed induction of aquaporin-1 (AQP1) during Staphylococcus aureus infection of cultured cells indicating a potential mechanism underlying altered water homeostasis during infection. To investigate mechanisms of AQP1 induction, we examined the effects of the S. aureus α-hemolysin on AQP1 abundance in Balb/c fibroblasts. Fibroblasts incubated with 30 µg/ml hemolysin exhibited a 5-10 fold increase in AQP1 protein within 4-6 hours of exposure. The use of multiple signaling cascade inhibitors failed to affect hemolysin-mediated accumulation of AQP1. However, immunoprecipitation revealed an initial accumulation of ubiquitinated AQP1 followed by a decrease to baseline levels after 4 hours. Immunofluorescence indicated that following hemolysin exposure, AQP1 was no longer on the plasma membrane, but was found in a population of submembrane vacuoles. AQP1 redistribution was further indicated by surface biotinylation experiments suggesting diminished AQP1 abundance on the plasma membrane as well as redistribution out of lipid raft fractions. Live cell confocal microscopy revealed that the pattern of cell volume change observed following hemolysin exposure was altered in cells in which AQP1 was silenced. We conclude that alpha-toxin alters proteasomal processing and leads to intracellular accumulation of AQP1, which may likely contribute to disrupted cell volume homeostasis in infection.
Keywords Aquaporin, Toxin, Ubiquitin, Lipid raft
Address and Contact Information 1Department of Medicine, Division of Pulmonary and Critical Care Medicine Johns Hopkins University School of Medicine, 5501 Hopkins Bayview Circle Baltimore, MD 21224, USA,
2Institute of Respiratory Disease, China Medical University, Shenyang, China
*Author for correspondence; e-mail: lsking@jhmi.edu, tel: 443-287-3347, fax: 410-287- 3349
[Rozmiar: 1332 bajtów]

DOI: 10.2478/s11658-007-0048-4 Volume 13 (2008) pp 212-229
Authors Benedykt Władyka* and Katarzyna Pustelny
Abstract Proteases, also referred to as peptidases, are the enzymes that catalyse the hydrolysis of peptide bonds in polipeptides. A variety of biological functions and processes depend on their activity. Regardless of the organism’s complexity, peptidases are essential at every stage of life of every individual cell, since all protein molecules produced must be proteolytically processed and eventually recycled. Protease inhibitors play a crucial role in the required strict and multilevel control of the activity of proteases involved in processes conditioning both the physiological and pathophysiological functioning of an organism, as well as in host-pathogen interactions. This review describes the regulation of activity of bacterial proteases produced by dangerous human pathogens, focusing on the Staphylococcus genus.
Keywords Protease, Protease inhibitor, Zymogen, Operon, Staphylococcus
Address and Contact Information Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Kraków, Poland
# Paper authored by participants of the international conference: XXXIV Winter School of the Faculty of Biochemistry, Biophysics and Biotechnology of Jagiellonian University, Zakopane, March 7-11, 2007, "The Cell and Its Environment". Publication cost was partially covered by the organisers of this meeting.
* Author for correspondence; e-mail: wladykab@interia.pl, tel: +48 12 664 6506, fax: +48 12 664 6915
[Rozmiar: 1332 bajtów]

DOI: 10.2478/s11658-007-0049-3 Volume 13 (2008) pp 230-239
Authors Jan Kowalski1, Maciej Banach2*, Marcin Barylski1, Robert Irzmanski1 and Lucjan Pawlicki1
Abstract The aim of this study was to evaluate the effect of carvedilol on the enzymatic antioxidative defence and plasma antioxidative activity in patients with stable angina. The study comprised 30 patients, aged 37-49 years with stable angina. Patients received carvedilol in escalating doses of 12.5 mg/24 h, 25 mg/24 h, and 50 mg/24 h for 4 weeks each. The control group was matched for age and gender, and consisted of 12 healthy volunteers, aged 39-49 years. Blood samples were collected from the cubital vein before and 4, 8 and 12 weeks after the therapy from the patients and once from the control group. For all the subjects, the superoxide dismutase (SOD-1), glutathione peroxidase (GSH-Px), catalase (CAT) activities in the erythrocytes and the antioxidant activity of the blood plasma were determined. The enzymatic antioxidative defence was significantly decreased in patients with stable angina in comparison to the healthy subjects. During the carvedilol therapy, an increase in the SOD-1, GSH-Px and CAT activities was observed. Moreover, 8 and 12 weeks after carvedilol therapy, the GSH-Px activity did not differ significantly from that observed in the group of healthy subjects. Carvedilol also increased the plasma antioxidative activity in patients with stable angina, but its level remained significantly lower than in the control group. In conclusion, carvedilol enhances antioxidant defense mechanisms in patients with chronic stable angina pectoris.
Keywords Stable angina, Carvedilol, Superoxide dismutase, Peroxidase, Catalase, Plasma antioxidative activity
Address and Contact Information 1Department of Internal Medicine and Cardiological Rehabilitation, Medical University of Łódź, Poland,
2Department of Cardiology, 1st Chair of Cardiology and Cardiac Surgery, Medical University of Łódź, Poland
* Author for correspondence: Department of Cardiology, 1st Chair of Cardiology and Cardiac Surgery, Medical University in Łódź, Poland. Sterlinga 1/3; 91-425 Łódź, Poland, e-mail: m.banach@termedia.pl , tel/fax: +48 42 636 44 71
[Rozmiar: 1332 bajtów]

DOI: 10.2478/s11658-007-0050-x Volume 13 (2008) pp 240-249
Authors Kulthum Mohammed and Amal Shervington٭
Abstract Previously, we identified a novel correlation between the upregulated expression of telomerase (hTERT) and cytochrome P450 1A1 (CYP1A1) in A549 human lung cancer cell line. The expression correlation was confirmed by silencing CYP1A1 expression using siRNA technology and observing a silencing of hTERT transcription. Furthermore, silencing CYP1A1 and subsequently downregulating hTERT resulted in the reduction of cancer cell viability by more than 40%, which appeared as early as 24 hours after the treatment. The concomitant downregulation of CYP1A1 and hTERT resulted in rapid cell death. This finding can be further exploited to develop new molecular targets for the treatment of lung cancer.
Keywords siRNA, Gene knockdown, CYP1A1, hTERT, Transfection
Address and Contact Information University of Central Lancashire, Department of Biological Sciences, Preston, PR1 2HE, United Kingdom
*Author for correspondence; e-mail: aashervington@uclan.ac.uk, tel.: +44(0)1772 893598
[Rozmiar: 1332 bajtów]

DOI: 10.2478/s11658-007-0052-8 Volume 13 (2008) pp 250-259
Authors An Claessens, Christine Weyn and Joseph Merregaert*
Abstract Mouse chondrolectin (chodl) was isolated out of the tail tip of fourday old 129/SvJ mice as a by-product of a PCR-based subtractive cDNA library screening. The gene is predominantly expressed in adult skeletal muscle, heart, testes and lungs and in embryonic stadia. Chodl is the mouse homologue of human chondrolectin (CHODL), a gene that encodes for a type Ia transmembrane protein and that is expressed in human testis, prostate, heart and skeletal muscle tissue. CHODL-splice variants (CHODLf, CHODLfΔE, CHODLΔE) are detected in human leukocytes. The proteins of the chondrolectin family belong to the family of C-type lectins. As the members of this protein family are important for a wide array of biological processes, the function of chodl was investigated by searching for its protein interaction partners. The ß-subunit of Rab geranylgeranyl transferase (Rabggtb) was isolated 8 times after a complete Sos recruitment system (SRS) screen with the cytoplasmic domain of chodl. The interaction was confirmed with in vitro transcription/translation and co-immunoprecipitation (co-IP) experiments.
Keywords C-type lectin, Chondrolectin, SRS
Address and Contact Information Laboratory of Molecular Biotechnology, Department of Biomedical Sciences, University of Antwerp, Universiteitsplein 1, B-2610 Wilrijk, Belgium
* Author for correspondence; e-mail: joseph.merregaert@ua.ac.be, tel.: 32-3-820-2311, fax: 32-3-820-2248.
[Rozmiar: 1332 bajtów]

DOI: 10.2478/s11658-007-0054-6 Volume 13 (2008) pp 260-270
Authors David H. Craig1,4, Keri L. Schaubert4, Hiroe Shiratsuchi1, June Kan-Mitchell4, and Marc D. Basson1,2,3,4*
Abstract Patients with malignancy typically exhibit abnormal dendritic cell profiles. Interstitial tumor pressure is increased 20-50mmHg over that in normal tissue. We hypothesized that elevated pressure in the tumor microenvironment may influence dendritic cell (DC) phenotype and function. Monocyte-derived immature and mature DC isolated from healthy human donors were exposed to either ambient or 40 mmHg increased pressure at 37°C for 12 hours, then assessed for expression of CD80, CD86, CD83, CD40, MHC-I and MHC-II. IL-12 production and phagocytosis of CFSE-labeled tumor lysate were assessed in parallel. Elevated pressure significantly increased expression of all co-stimulatory and MHC molecules on mature DC. Immature DC significantly increased expression of CD80, CD86, CD83 and MHC-II, but not MHC-I and CD40, versus ambient pressure controls. Pressure-treated immature DC phenotypically resembled mature DC controls, but produced low IL-12. Phenotypic maturation correlated with decreased phagocytic capacity. These results suggest increased extracellular pressure may cause aberrant DC maturation and impair tumor immunosurveillance.
Keywords Pressure, Dendritic cell, Maturation, Cancer, Immunosurveillance
Address and Contact Information 1Departments of Surgery, 2Anesthesiology, 3Anatomy and Cell Biology, 4Karmanos Cancer Institute, John D. Dingell VA Medical Center and Wayne State University, 4646 John R. Street, Detroit, MI 48201, USA
*Author for correspondence; e-mail: marc.basson@va.gov, tel.: 313-576-3598, fax: 313-576-1002
[Rozmiar: 1332 bajtów]

DOI: 10.2478/s11658-007-0055-5 Volume 13 (2008) pp 271-282
Authors Ewa Kowalska* and Andrzej Kozik
Abstract Thiamin (vitamin B1) is an essential molecule for all living organisms. Its major biologically active derivative is thiamin diphosphate, which serves as a cofactor for several enzymes involved in carbohydrate and amino acid metabolism. Important new functions for thiamin and its phosphate esters have recently been suggested, e.g. in gene expression regulation by influencing mRNA structure, in DNA repair after UV illumination, and in the protection of some organelles against reactive oxygen species. Unlike higher animals, which rely on nutritional thiamin intake, yeasts can synthesize thiamin de novo. The biosynthesis pathways include the separate synthesis of two precursors, 4-amino -5-hydroxymethyl-2-methylpyrimidine diphosphate and 5-(2-hydroxyethyl)- 4-methylthiazole phosphate, which are then condensed into thiamin monophosphate. Additionally, yeasts evolved salvage mechanisms to utilize thiamin and its dephosphorylated late precursors, 4-amino-5-hydroxymethyl- 2-methylpyrimidine and 5-(2-hydroxyethyl)-4-methylthiazole, from the environment. The current state of knowledge on the discrete steps of thiamin biosynthesis in yeasts is far from satisfactory; many intermediates are postulated only by analogy to the much better understood biosynthesis process in bacteria. On the other hand, the genetic mechanisms regulating thiamin biosynthesis in yeasts are currently under extensive exploration. Only recently, the structures of some of the yeast enzymes involved in thiamin biosynthesis, such as thiamin diphosphokinase and thiazole synthase, were determined at the atomic resolution, and mechanistic proposals for the catalysis of particular biosynthetic steps started to emerge.
Keywords Thiamin biosynthesis, Thiamin diphosphate, Thiazole, Pyrimidine, THI genes, Saccharomyces cerevisiae
Address and Contact Information Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Kraków, Poland
* Author for correspondence; e-mail: ewa.b.kowalska@uj.edu.pl, tel.: (4812) 664 65 44, fax: (4812) 664 69 02
# Paper authored by participants of the international conference: XXXIV Winter School of the Faculty of Biochemistry, Biophysics and Biotechnology of Jagiellonian University, Zakopane, March 7-11, 2007, "The Cell and Its Environment". Publication cost was partially covered by the organisers of this meeting.
[Rozmiar: 1332 bajtów]

DOI: 10.2478/s11658-008-0001-1 Volume 13 (2008) pp 283-302
Authors Jie-Ying Liao1, James Q. Yin1*, Fang Chen1, Tie-Gang Liu2 and Jia-Chang Yue1*
Abstract Although there are many reports about the efficacy of siRNAs, it is not clear whether those siRNAs with high C/G contents can be used to silence their target mRNAs efficiently. In this study, we investigated the structure and function of a group of siRNAs with high C/G contents. The results showed that single siRNAs against the Calpain, Otoferlin and Her2 mRNAs could induce different silencing effects on their targets, suggesting that the accessibility to target sequences influences the efficacy of siRNA. Unexpectedly, a single siRNA could target its cognate sequence in the 3’UTR of EEF1D or the 5’UTR of hTRF2 or CDC6. Their interaction induced different modes of gene silencing. Furthermore, the introduction of mutations into the 3’ end of the passenger strand showed that the position and number of mutated nucleotides could exert some influence on the efficacy of siRNA. However, these mutations did not completely block the passenger strand from exerting its RNAi effect. Interestingly, our findings also indicated that the target mRNA might play essential roles in maintaining or discarding the guide strand in RISCs. Thus, the conclusion could be drawn that favorable siRNA sequences, accessible target structures and the fast cleavage mode are necessary and sufficient prerequisites for efficient RNAi.
Keywords siRNA, RNAi, mRNA, Local structure, Gene silencing
Address and Contact Information 1Systems Biology Research Center, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101, P.R. China
2Molecular Oncology Research Institute, Tufts-NEMC, Boston, MA 02111, USA
*Authors for correspondence. James Q. Yin, e-mail: jqwyin@sun5.ibp.ac.cn, tel.: 86-010-64888572, fax: 86-010-64888572 or Jiachang Yue, e-mail: yuejc@sun5.ibp.ac.cn
[Rozmiar: 1332 bajtów]

DOI: 10.2478/s11658-008-0002-0 Volume 13 (2008) pp 303-311
Authors Jia Huang1, Shu-Jing Li1, Xian-Hua Chen2*, Yu Han and Ping Xu2*
Abstract This study focused on the function of hnRNP-R in the regulation of c-fos expression. We demonstrated that hnRNP-R accelerated the rise and decline phases of c-fos mRNAs and Fos proteins, allowing PMA to induce an augmented pulse response of c-fos expression. Then, we examined the role of the c-fos-derived AU-rich element (ARE) in hnRNP-R-regulated mRNA degradation. Studies with the ARE-GFP reporter gene showed that hnRNP-R significantly reduced the expression of GFP with an inserted ARE. Moreover, immunoprecipitation-RT-PCR analysis demonstrated that in R28 cells and rat retinal tissues, the c-fos mRNA was co-immunoprecipitated with hnRNP-R. These findings indicate that hnRNP-R regulates the c-fos expression in retinal cells, and that the ARE of c-fos mRNAs contributes to this regulation.
Keywords hnRNP-R, Retina, c-fos, mRNA turnover, ARE
Address and Contact Information 1Laboratory of Genomic Physiology and 2State Key Laboratory of Medical Neurobiology, Fudan University, 138 Yixueyuan Road, Shanghai, 200032, P. R. China
*Authors for correspondence; e-mail: matibuck@yahoo.com or xhchen@fudan.edu.cn, tel: 086-21-54237094, fax: 086-21-54237094
[Rozmiar: 1332 bajtów]

DOI: 10.2478/s11658-008-0004-y Volume 13 (2008) pp 312-326
Authors Włodzimierz Korohoda* and Anna Wilk
Abstract In this paper, we discuss the application of various methods of cell electrophoresis in research into cell surface properties (analytical methods), and the separation of uniform cell subpopulations from cell mixtures (preparative methods). The emphasis is on the prospects of the development of simplified and versatile methodologies, i.e. microcapillary cell electrophoresis and horizontal cell electrophoresis under near-isopycnic conditions. New perspectives are considered on the use of analytical and preparative cell electrophoresis in research on cell differentiation, neoplastic transformation, cell-cell interactions and the biology of stem cells.
Keywords Cell electrophoresis, Cell separation, Cell surface
Address and Contact Information Department of Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, ul. Gronostajowa 7, 30-387 Kraków, Poland
# Paper authored by participants of the international conference: XXXIV Winter School of the Faculty of Biochemistry, Biophysics and Biotechnology of Jagiellonian University, Zakopane, March 7-11, 2007, "The Cell and Its Environment". Publication cost was covered by the organisers of this meeting.
*Author for correspondence; e-mail: korohoda@moj.uj.edu.pl
[Rozmiar: 1332 bajtów]