Cellular & Molecular Biology Letters
International Scientific Journal
Established 1996
Volume 10 (2005) No. 3
| Volume 10 (2005) pp 373-382 | |
| Title | 31P MRS ANALYSIS OF THE PHOSPHOLIPID COMPOSITION OF NORMAL HUMAN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMC) |
| Authors | Ma³gorzata Kuliszkiewicz-Janus1*, Mariusz Adam Tuz2, Stanis³aw Baczyñski3, Iwona Prajs1 and Bożena Jaĵwiec1 |
| Abstract | The aim of this investigation was to characterize the phospholipid composition of normal human blood mononuclear cells using 31P NMR spectroscopy. Mononuclear cells of peripheral blood were obtained from 10 volunteers. Phospholipid extracts were prepared from 60´106 cells according to modified Folch’s method. An AMX 300 Bruker spectrometer 7.05 T was used. The 31P spectrum of phospholipid extracts from normal human PBMC consisted of 9 peaks, with one each for phosphatidylcholine (PC), plasmalogen of phosphatidylcholine (CPLAS), lysophosphatidylcholine (LPC), sphingomyelin (SM), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS) and cardiolipin (CL), and another one due to the external reference substance, methylenediphosphonic acid (MDPA). The concentrations of these phospholipids (PL), based on the integral intensities, were as follows: 0.398ħ0.078 mmole/l for PC; 0.033ħ0.019 mmole/l for CPLAS; 0.155ħ0.043 mmole/l for SM; 0.266ħ0.104 mmole/l for PI+PE; 0.101ħ0.040 mmole/l for PS, and 0.026ħ0.033 mmole/l for CL. The results of this study confirmed that 31P MRS is a convenient tool for measuring the phospholipid concentrations of biological samples. |
| Adress and Contact Informations | 1Department. of Hematology, Wroc³aw Medical University, Poland, 2Institute of Experimental Physics, University of Wroc³aw, Poland, 3Institute of Chemistry, University of Wroc³aw, Poland * Corresponding author; e-mail: mkj@hemat.am.wroc.pl |
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| Volume 10 (2005) pp 383-388 | |
| Title | ENHANCED PRODUCTION OF ISOAMYL ALCOHOL AND ISOAMYL ACETATE BY UBIQUITINATION-DEFICIENT Saccharomyces cerevisiae MUTANTS # |
| Authors | Fumiyoshi Abe* and Koki Horikoshi |
| Abstract | Aromatic compounds are an important element in the flavor of yeastfermented alcohol. We isolated mutants of Saccharomyces cerevisiae capable of growth at high levels of hydrostatic pressure. Among them, the HPG1 mutants, with a defect in their Rsp5 ubiquitin ligase, were found to produce high amounts of aromatics due to enhanced leucine uptake, with isoamyl alcohol production 2- to 3-fold and isoamyl acetate production 4- to 8-fold that of the wild-type strain. The result suggests that the HPG1/RSP5 mutant alleles could be new resources for producing these flavoring compounds for yeast-fermented alcoholic beverages. |
| Adress and Contact Informations | Extremobiosphere Research Center, Japan Agency for Marine-Earth Science and
Technology, 2-15 Natsushima-cho, Yokosuka 237-0061, Japan # Invited paper * Corresponding author; e-mail: abef@jamstec.go.jp |
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| Volume 10 (2005) pp 389-400 | |
| Title | THE INFLUENCE OF METHYLENE BLUE ON THE SPONTANEOUS CONTRACTITY OF THE NON-PREGNANT HUMAN MYOMETRIUM AND ON THE MYOMETRIAL RESPONSE TO DEA/NO |
| Authors | Beata Modzelewska* and Anna Kostrzewska |
| Abstract | Nitric oxide (NO) is a potent inhibitor of spontaneous contractions of the human non-pregnant myometrium; however, the precise mechanism by which NO causes the myometrial smooth muscles to relax remains unclear. The aim of this study was to determine the influence of methylene blue (MB) on myometrial contractions and the response of the myometrium to DEA/NO in vitro. Concentration-response curves to DEA/NO were constructed in the absence and presence of MB (5·10-6, 10-4 and 10-2 mol/l) and 5·10-3 mol/l cystamine. Cystamine did not counteract the DEA/NO-induced relaxation of the myometrial strips. MB itself, excluding the lowest concentration, caused noticeable changes in spontaneous activity. The changes involved a concentration-dependent increase in the frequency of contractions, and a decrease in their amplitude. In conclusion, our results confirm that NO relaxes the human myometrium via a cGMP-independent mechanism. The results obtained in the presence of MB may be misleading because of its complex influence on myometrial contractile activity |
| Adress and Contact Informations | Department of Biophysics, Medical University of Bia³ystok, ul. Mickiewicza
2A, 15-230 Bia³ystok, Poland * Corresponding author, e-mail: beamo@amb.edu.pl |
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| Volume 10 (2005) pp 401-411 | |
| Title | CLONING AND CHARACTERIZATION OF cDNA FOR THE Oryza sativa PHOSPHATE TRANSPORTER |
| Authors | Feng Ming1,2*, Guo Hua Mi3, Qun Lu1,2, Sai Yin1,2, Shan Shan Zhang1,2, Bin Guo1,2 and Da Leng Shen1,2 |
| Abstract | A putative high-affinity phosphate (Pi) transporter gene in rice (Oryza sativa), OsLPT1, was isolated by RT-PCR from the leaves of the plants. The 1635-bp nucleotide sequence of OsLPT1 spans an open reading frame encoding a polypeptide of 535 amino acids with sequence similarity to phosphate transporters from other plant species. Southern blot analysis showed that the OsLPT1 gene might be present in three transcripts in the rice genome. RT-PCR analysis demonstrated the expression of OsLPT1 in both leaves and roots. The expression of OsLPT1 in the roots was enhanced by Pi deprivation. In situ hybridization revealed OsLPT1 expression in mesophyll cells, xylem parenchyma and phloem cells in the leaves, and in the epidermis, exodermis, and in the vasculature surrounding metaxylem vessels in the roots. The data suggests that the OsLPT1 protein may be involved in enhancing phosphate uptake under conditions of Pi starvation, and in the translocation of Pi among cells in shoots to increase the efficiency of internal Pi use. |
| Adress and Contact Informations | 1Institute of Genetics, State Key Laboratory of Genetic Engineering, 2 Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, Institute of Biodiversity Science, School of Life Science, Fudan University 220 Handan Road, Shanghai 200433, P.R. China, 3 Plant Nutrition Department of China Agricultural University, Beijing, 100094, P.R. China * Corresponding author; tel: +86-21-65643331, fax: +86-21-65648376, e-mail: fming@fudan.edu.cn |
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| Volume 10 (2005) pp 413-423 | |
| Title | LEAD-INDUCED CELL DEATH OF HUMAN NEUROBLASTOMA CELLS INVOLVES GSH DEPRIVATION |
| Authors | Chellu S. Chetty1*, Mohan C. Vemuri2, Khamisi Campbell1 and Challa Suresh3 |
| Abstract | Lead (Pb2+) is a toxic heavy metal that has adverse effects on the health of humans and other animals. The developing central nervous system is especially sensitive and vulnerable to Pb2+ toxicity. In this study, the effects of low levels of Pb2+ exposure on human SH-SY5Y neuroblastoma cell cultures were assessed. The cells were exposed to Pb2+ (0.01 mM - 10 mM) for 48 hrs, and the level of cell proliferation was determined. Pb2+ significantly inhibited the proliferation of neuroblastoma cells in a concentration-dependent manner. A 50% inhibition (IC50) in cellular proliferation was observed with 5 mM Pb2+. A significant decrease in the levels of glutathione (GSH), a critical intracellular antioxidant, was observed at all the lead concentrations. There was also a multifold increase in the activity of caspase-3, a key executioner of apoptosis, and in the levels of prostaglandin E2 (PGE2). Our results suggest that the neurotoxic effects of Pb may be mediated by apoptosis and PGE2 release, which could be potentially detrimental to neuronal survival. |
| Adress and Contact Informations | 1Department of Natural Sciences and Mathematics, Savannah State University,
Savannah, GA 31404, USA, 2Department of Surgery, Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USA, 3National Institute of Nutrition, Hyderabad, AP 500 007, India * Corresponding author; tel: 912-353-3057, fax: 912-303-4376, e-mail: chettyc@savstate.edu |
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| Volume 10 (2005) pp 425-437 | |
| Title | CROSS-SPECIES BACTERIAL ARTIFICIAL CHROMOSOME (BAC) LIBRARY SCREENING VIA OVERGO-BASED HYBRIDIZATION AND BAC-CONTIG MAPPING OF A YIELD ENHANCEMENT QUANTITATIVE TRAIT LOCUS (QTL) YLD1.1 IN THE MALAYSIAN WILD RICE Oryza rufipogon |
| Authors | Beng-Kah Song1, Kalaivani Nadarajah2*, Michael N. Romanov3 and Wickneswari Ratnam1 |
| Abstract | The construction of BAC-contig physical maps is an important step towards a partial or ultimate genome sequence analysis. Here, we describe our initial efforts to apply an overgo approach to screen a BAC library of the Malaysian wild rice species, Oryza rufipogon. Overgo design is based on repetitive element masking and sequence uniqueness, and uses short probes (~40 bp), making this method highly efficient and specific. Pairs of 24-bp oligos that contain an 8-bp overlap were developed from the publicly available genomic sequences of the cultivated rice, O. sativa, to generate 20 overgo probes for a 1- Mb region that encompasses a yield enhancement QTL yld1.1 in O. rufipogon. The advantages of a high similarity in melting temperature, hybridization kinetics and specific activities of overgos further enabled a pooling strategy for library screening by filter hybridization. Two pools of ten overgos each were hybridized to high-density filters representing the O. rufipogon genomic BAC library. These screening tests succeeded in providing 69 PCR-verified positive hits from a total of 23,040 BAC clones of the entire O. rufipogon library. A minimal tilling path of clones was generated to contribute to a fully covered BAC-contig map of the targeted 1-Mb region. The developed protocol for overgo design based on O. sativa sequences as a comparative genomic framework, and the pooled overgo hybridization screening technique are suitable means for high-resolution physical mapping and the identification of BAC candidates for sequencing. |
| Adress and Contact Informations | 1School of Environmental and Natural Resource Sciences, Faculty of Science
and Technology, Kebangsaan University Malaysia, 43600 Bangi, Selangor,
Malaysia; 2School of BioScience and Biotechnology, Faculty of Science and Technology, Kebangsaan University Malaysia, 43600 Bangi, Selangor, Malaysia; 3Genetics Division, CRES (Conservation and Research for Endangered Species), Zoological Society of San Diego, Arnold and Mabel Beckman Center for Conservation Research, 15600 San Pasqual Valley Road, Escondido, CA 92027-7000, USA * Corresponding author; tel.: +60-3-89213465, fax: +60-3-89252698, e-mail: vani@pkrisc.cc.ukm.my |
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| Volume 10 (2005) pp 439-453 | |
| Title | THE SUBCELLULAR DISTRIBUTION OF THE p53 TUMOUR SUPPRESSOR, AND ORGANISMAL AGEING # |
| Authors | Józefa Wêsierska-Għdek*1 and Gerald Schmid1,2 |
| Abstract | The p53 protein, the product of a tumour suppressor gene, is a key regulator of cell growth, differentiation and apoptosis. It is able to induce a transient cell cycle arrest and terminal senescence. Most of its functions are exerted by the transcriptional activation of genes involved in cell cycle control, DNA repair and apoptosis. The activation of p53 is primarily mediated by posttranslational modifications that affect its conformation and capacity to bind to several proteins, resulting in its stabilization and enhanced DNA-binding potential. Another way to regulate the biological function of p53 involves changes in its intracellular distribution. This paper presents an overview of the role of p53 in cellular senescence and the regulation of p53 activity by its intracellular distribution. |
| Adress and Contact Informations | 1Cell Cycle Regulation Group, Div.: Institute of Cancer Research, Dept. of
Medicine I, Vienna Medical University, Borschkegasse 8 a, A-1090 Vienna,
Austria, 2LMU Klinikum Grosshadern; Experimentelle Forschung Chirurgie, Muenchen, Germany # Invited paper * Corresponding author, tel: (+43-1) 4277-65247, fax: (+43-1) 4277-65194, e-mail: Jozefa.Gadek-Wesierski@meduniwien.ac.at. |
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| Volume 10 (2005) pp 455-470 | |
| Title | SERUM-RESISTANT GENE TRANSFER TO ORAL CANCER CELLS BY METAFECTENE AND GENEJAMMER: APPLICATION TO HSV-tk/GANCICLOVIR-MEDIATED CYTOTOXICITY |
| Authors | Krystyna Konopka*, Basma Fallah, Jomarie Monzonduller, Nathan Overlid and Nejat Duzgunes |
| Abstract | Cationic lipids and polyamines have been used as non-viral gene transfer reagents, both in vitro and in vivo. One of the limitations to their use in vivo is the inhibition of gene delivery by serum. We showed previously that, in the absence of serum, relatively high cytotoxicity in oral cancer cell lines could be achieved via transfection of the Herpes Simplex Virus thymidine kinase (HSV-tk) gene followed by treatment with ganciclovir (GCV), despite the low efficiency of transfection (Konopka et al., Gene Ther. Mol. Biol. 8 (2004) 307- 318). In this study we evaluated the effect of high concentrations (20-60%) of fetal bovine serum (FBS) on the transfection efficiency of two novel reagents, the polycationic liposome, Metafectene, and the polyamine reagent, GeneJammer, in HSC-3 and H357 human oral squamous cell carcinoma (OSCC) cells. We also examined whether the HSV-tk gene delivered in the presence of FBS (up to 60%), could induce cell death following treatment with GCV. Transfection was optimized using a luciferase-expressing plasmid. Both Metafectene- and GeneJammer-mediated luciferase gene expression in HSC-3 cells was reduced by 40-50% when transfection was performed in the presence of 20-60% FBS. The delivery of the HSV-tk gene by Metafectene in the absence and the presence of 60% FBS, followed by GCV treatment for 9 days, resulted in 95% and 70% cytotoxicity, respectively. With GeneJammer, transfection in 0% and 60% FBS resulted in 90% and 40% cytotoxicity, respectively, after 9 days. In contrast, very low transfection activity and a much higher inhibitory effect of serum were observed in H357 cells. Nevertheless, about 35% GCV mediated cytotoxicity was observed with H357 cells at both 0% and 60% FBS, using GeneJamer. Thus, Metafectene and GeneJammer can be used in the delivery of genes in biological milieu and in the gene therapy of OSCC in animal models. |
| Adress and Contact Informations | Department of Microbiology, University of the Pacific, Arthur A. Dugoni
School of Dentistry, 2155 Webster Street, San Francisco, CA 94115, USA * Corresponding author; e-mail: kkonopka@pacific.edu |
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| Volume 10 (2005) pp 471-478 | |
| Title | CHANGES IN THE ACTIVITY OF ACETYLCHOLINESTERASE AND NA,K-ATPASE IN HUMAN ERYTHROCYTES IRRADIATED WITH X-RAYS |
| Authors | Anita Krokosz* and Zofia Szweda-Lewandowska |
| Abstract | The response of human erythrocytes to X-rays in the dose range from 40 Gy to 600 Gy was determined on the basis of changes in the activities of AChE and ATPase. The Na,K-ATPase activity increased above the control value at doses below 200 Gy, while at the doses higher than 200 Gy, it decreased, reaching 96% of the control value at a dose of 600 Gy. In the range of doses up to 200 Gy, the AChE activity, expressed as Vmax, did not change. At higher doses, it fell drastically, reaching 33% of the control value at a dose of 600 Gy. Simultaneously, the enzyme substrate affinity decreased at 200 Gy, and then started to increase at lower values of Vmax. The obtained results suggest that under appropriate conditions, low doses of radiation may have the opposite effects to high doses. |
| Adress and Contact Informations | Department of Molecular Biophysics, University of £ódĵ, ul. Banacha 12/16,
90-237 £ódĵ, Poland * Corresponding author; e-mail: krokosz@biol.uni.lodz.pl |
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| Volume 10 (2005) pp 479-498 | |
| Title | THE MAMMALIAN TARGET OF THE RAPAMYCIN (mTOR) KINASE PATHWAY: ITS ROLE IN TUMOURIGENESIS AND TARGETED ANTITUMOUR THERAPY |
| Authors | Agnieszka Janus, Tadeusz Robak and Piotr Smolewski* |
| Abstract | The mammalian target of rapamycin (mTOR) is a kinase responsible for mitogen-induced cell proliferation/survival signaling. Its activation in response to mitogens leads to a cell-cycle progression from G1 to S phase. mTOR controls the acivation of ribosomal protein translation and the initiation of cap-dependent translation. A role of mTOR signaling pathway dysregulation in tumourigenesis is postulated. mTOR and pathways upstream of this kinase were found to be frequently upregulated in neoplastic diseases. Therefore, it is also an attractive target for antitumour therapy. Several mTOR inhibitors were developed, including rapamycin and its analogues: CCI-779, RAD001 and AP23573. After promising phase I studies, their potential clinical significance is currently under evaluation in several phase II-III trials on patients with solid tumours and some hematological malignancies. |
| Adress and Contact Informations | Department of Hematology, Medical University of £ódĵ, Copernicus Memorial
Hospital, Cio³kowskiego 2, 93-510 £ódĵ, Poland * Corresponding author: tel: +48-42-689-51-91, fax: +48-42-689-51-92; e-mail: piotr_smolewski@wp.pl |
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| Volume 10 (2005) pp 499-514 | |
| Title | CONSTRUCTION OF A STABLE CELL LINE UNIFORMLY EXPRESSING THE RAT TRPV1 RECEPTOR |
| Authors | Zoltan Sandor1, Angelika Varga1, Peter Horvath2, Barbara Nagy3 And Janos Szolcsanyi1* |
| Abstract | We constructed and analyzed a new cell line called HT5-1, which stably expresses an enhanced green fluorescent protein-tagged version of the rat vanilloid receptor 1 (VR1/TRPV1). The fluorescent receptor allowed easy measurement of receptor expression and expression level-based purification of cells via fluorescence-activated cell sorting. The HT5-1 cell line was compared to cells transiently transfected with the fluorescent receptor, to cells expressing the native rat vanilloid receptor, and to isolated capsaicin-sensitive rat trigeminal sensory neurons. Fura-2 microfluorimetry measurements of the calcium influx upon capsaicin induction showed that, by contrast to transiently transfected cells, HT5-1 cells respond uniformly to the stimulation, due to the similar level of receptor expression in individual cells. HT5-1 cells showed similar behaviour to isolated trigeminal root ganglion neurons, including marked tachyphylaxis upon repeated capsaicin induction, and a lack of calcium ion release from intracellular storage sites. |
| Adress and Contact Informations | 1Department of Pharmacology and Pharmacotherapy, Faculty of Medicine,
Neuropharmacology Research Group of the Hungarian Academy of Sciences,
University of Pécs, Szigeti u. 12, H-7624 Pécs, Hungary, 2Department of Pharmacology and Pharmacotherapy, Medical and Health Science Center, University of Debrecen, Nagyerdei krt. 98, H-4032, Debrecen, Hungary, 3Department of Biophysics, Faculty of Medicine, University of Pécs, Szigeti u. 12, H-7624 Pécs, Hungary * Corresponding author; tel: 36-72-536217, fax: 36-72-536218, e-mail: janos.szolcsanyi@aok.pte.hu |
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| Volume 10 (2005) pp 515-534 | |
| Title | THE CORRELATION BETWEEN OXIDATIVE STRESS AND LEAF SENESCENCE DURING PLANT DEVELOPMENT # |
| Authors | Petra Zimmermann and Ulrike Zentgraf* |
| Abstract | In plants, besides being the final step leading to the death of the whole organism, senescence has a developmental function involving the coordinated degradation of macromolecules and the mobilization of nutrients out of senescing tissues into developing parts of the plant. Free radicals are thought to play an essential role in senescence, especially those derived from oxygen. Since these molecules are extremely toxic, the levels of the different reactive oxygen species have to be tightly regulated. However, at low concentrations, hydrogen peroxide may also serve as a signalling molecule. Therefore, a coordinated regulation of the free radical scavenging system, which comprises enzymatic components such as catalase, superoxide dismutase and ascorbate peroxidase, and non-enzymatic molecules such as ascorbate and glutathione is essential. The increased radical levels displayed during senescence are not only caused by the elevated production of radicals but also by a loss in antioxidant capacity. |
| Adress and Contact Informations | ZMBP, Centre of Plant Molecular Biology, University of Tübingen,
Auf der Morgenstelle 28, 72076 Tübingen, Germany # Invited paper * Corresponding author; e-mail: ulrike.zentgraf@uni-tuebingen.de |
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| Volume 10 (2005) pp 535-551 | |
| Title | RAT COLONIC LIPID PEROXIDATION AND ANTIOXIDANT STATUS: THE EFFECTS OF DIETARY LUTEOLIN ON 1,2- DIMETHYLHYDRAZINE CHALLENGE |
| Authors | Vaiyapuri Manju, Vairappan Balasubramaniyan and Namasivayam Nalini* |
| Abstract | Colon cancer is the third most common cancer and second leading cause of cancer-related death in the United States. A number of recent articles demonstrate the importance of natural products as cancer chemopreventive agents. In this study, we evaluated the chemopreventive efficacy of luteolin, a flavonoid, on tissue lipid peroxidation and antioxidant status, which are used as biomarkers in DMH-induced experimental colon carcinogenesis. Rats were given a weekly subcutaneous injection of DMH at a dose of 20 mg/kg body weight for 15 weeks. Luteolin (0.2 mg/kg body weight/everyday p.o.) was given to the DMH-treated rats at the initiation and post-initiation stages of carcinogenesis. The animals were killed after 30 weeks. After a total experimental period of 32 weeks (including 2 weeks of acclimatization), tumor incidence was 100% in DMH-treated rats. In those DMH-treated rats that had received luteolin during the initiation or post-initiation stages of colon carcinogenesis, the incidence of cancer and the colon tumor size was significantly reduced as compared to that for DMH-treated rats not receiving luteolin. In the presence of DMH, relative to the results for the control rats, there were decreased levels of lipid peroxidation, as denoted by thiobarbituric acid reactive substances (TBARS), conjugated dienes and lipid hydroperoxides, decreased activities of the enzymic antioxidants superoxide dismutase (SOD) and catalase (CAT), and elevated levels of glutathione and the glutathionedependent enzymes reduced glutathione (GSH), glutathione peroxidase (GPx), glutathione–S-transferase (GST) and glutathione reductase (GR), and of the nonenzymic antioxidants vitamin C and vitamin E. Our study shows that intragastric administration of luteolin inhibits colon carcinogenesis, not only by modulating lipid peroxidation and antioxidant status, but also by preventing DMH-induced histopathological changes. Our results thus indicate that luteolin could act as a potent chemopreventive agent for colon carcinogenesis. |
| Adress and Contact Informations | Department of Biochemistry, Faculty of Science,
Annamalai University, Annamalainagar – 608002, Tamilnadu, India * Corresponding author: tel.: 91- 4144-238343-227, e-mail: nalininam@yahoo.com |
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Office:
Cellular & Molecular Biology Letters
Faculty of Biotechnology
University of Wroc³aw
Przybyszewskiego 63/77
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Faculty of Biotechnology
University of Wroc³aw
Przybyszewskiego 63/77
51-148 Wroc³aw, Poland
fax:
+48 71 375 6208 or
+48 71 375 6234
+48 71 375 6234
e-mail:
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