Cell Journal (Yakhteh)
Year:2002 | Volume:3 | Issue:12|Papers Count:10

Introduction: Standard concentrations of antibiotics in culture media are thought to have no detectable toxic effects on the cultured cells. Since antibiotics are biologically active substances, the possibility that they interfere to some extent with cellular processes occurring in the cultured cells can not always be totally excluded. This study, therefore, was conducted to assess whether the presence of penicillin-streptomycin (pen-strep) during the IVM (In Vitro Maturation) of bovine COCs (Cumulus Oocyte Complexes) affect nuclear and cytoplasmic maturation and subsequent embryo development. Materials and Methods: Bovine COCs were cultured at 39oC in a humidified atmosphere with 5 % CO2 in air for 24 h in: 1- M199 supplemented with 10 % FCS, 0.05 IU/ml rhFSH and 100 units penicillin and 100 µg streptomycin/ ml. 2- M199 without FCS and rhFSH in the presence of pen-strep. Cultures without antibiotics served as control. Results: In vitro maturation of bovine COCs in the presence of pen-strep in culture medium supplemented with FCS and rhFSH significantly (P< 0.05) increased the percentage of MII oocytes. However, when the COCs were divided, on the basis of apperance of the cumulus investment, into bright and dark groups, the positive effect of pen-strep on nuclear maturation became less obvious in both types of COCs, 76% vs 72% in bright COCs (P= 0.149) and 83% vs 80% in dark COCs (P= 0.296) in treated and control groups, respectively. Moreover, when FCS and rhFSH were excluded from culture medium the positive effect of pen-strep on nuclear maturation (P= 0.402) was omitted. The percentage of oocytes with type III of cortical granules (CGs) distribution and the COCs expansion was not affected by the presence of pen-strep during IVM. The subsequent embryo development of IVM/IVF produced ova, wich were exposed to pen-strep during IVM, was significantly (P<0.05) decreased with respect to the blastocyst formation by day 9. Conclusion: The results indicate that the nuclear maturation of bovine oocytes was positively influenced by the presence of pen-strep during IVM when the culture media was supplemented with FCS and rhFSH. Moreover, despite of no notable effect of pen-strep on CGs distribution the subsequent embryo development was negatively influenced by the presence of pen-strep.

Introduction: Chitosan Possesses more than 5000 glucossamine residue. Since chitosan possesses potentially reactive amino functional groups, this polymer has been shown to be of potential use in many different fields, including as an antifungal compound in agriculture, flocculating in waste water treatment, a food additive and alimentary industries, hydrating agent in cosmetics and more recently, in biomedicine and several pharmaceutical preparations. More research is needed to study the effects of chitosan on the Escherichia coli. It is necessary to do more research on the effect of chitosan on the Escherichia cell. The purpose of the investigation was to determine the antibacterial effects of chitosan against Escherichia cell. Materials and Methods: Escherichia coli:AtCC 25992 and Eosine-Methylen Blue Agar was used .A standard of E.Coli (1000000 per ml) is incubated for 24 hourse in a series of tubes concentration. The effects of reaction temperature, pH value and salts on the inhibitory activity of chitosan against Escherichia Coli were investigated. Results: Chitosan had inhibitory effects on Escherichia coli activity (100% activity vs 1978%, P<0/05). Our results show that higher temperature and acidic pH increased the antibacterial effects of chitosan. Sodium ions might complex with chitosan and accordingly reduce chitosan activity against Escherichia Coli. Divalent cations reduced the antibacterial activity of chitosan. Conclusion: Our results show that higher temperature and acidic pH increased the antibacterial effects of chitosan. Sodium ions might complex with chitosan with chitosan and accordingly reduce activity againts Escherichia Coli. Monovalent and divalent cations might complex with chitosan and accordingly reduce chitosan activity against Escherichia Coli.

Introduction: Membrane cofactor protein (MCP or CD46) is one of the regulatory of complement system. It is present in all somatic cells except in RBC. It is also present in all body fluids and semen. The aim of this study was measuring the concentration of this protein in semen of fertile and infertile individuals and to study the relationship between the concentration of this protein and semen parameters. Materials and Methods: The man concentration of CD46 was measured by sandwich ELIZA in 8 normospermic, 22 asthenopsermic, 22 oligo-asthenospermic and 11 azoospermic individuals. In this technique one monoclonal antibody used for capture and another monoclonal antibody againts CD46 was used for detection. A polycolonal antibody conjugated to HRP was used for staining. Pearson correlation and t-test was used for statistical analysis. Results: The man concentrations were 710, 453, 582 and 639 ng/ml for normospermic, asthenospermic, oligo-asthenospermic and azoo sperm individuals-respectively. The pearson correlation showed a signification correlation between CD46 concentration with sperm motility (r=+0.59 & P<0.001), sperm viability (r=+0.53 & P<0.005) and sperm concentration (r=-0.37 & P<0.003). Conclusions: Previous reports suggest no difference between CD46 concentration between fertile and infertile individuals. However our results showed that the level of CD46 significantly reduces in asthenospermic and hyperspermic (>200 million/ml) individuals. The correlation between sperm motility and sperm concentration with CD46 suggests that CD46 might influence sperm motility and this molecule appears to be taken up by spermatozoa. Reduction in CD46 might predispose to become astheno, oligo or oligo-astheno-spermic.

Introduction: The morphology and ultrastructure of germinal epithelum Basement Memberane (BM) in patients with non-obstructive azoospermia was studied by means of transmision electron microscipy. Materials and Methods: 10 biopsies of testis tissue from patients with non-obstructive azoospermia referred to Royan Institute as patients group and 5 biopsies from patients with prostatic cancer as control group were used. After tissue proccessing for electron microscopy, the tissues were examined.Results: The most important result in this study, was thickening of BM. The other pathological changes were folding and invagination of BM toward germinal epithelum and multilayring and splitting (Delamination) of BM. Conclusion: The results showed that pathological changes were observed in all patients. Because th BM is made by both sertoli and myoid cells, maturation of these cells are important. Probably such pathological changes in BM are due to abnormality in these cells.

Introduction: Male infertility frequently occures after spinal cord injury (SCI). However little in known about the acute effect of SCI on male reproductive function. This study examined the spermatogenic changes during acute phase of SCI. Material and Metods: In experimental groups SCI was produced in male rats by T9 laminectomy and spinal cord transsection (N=23, Age=70 days). Control groups received simlar surgery, though, without laminectomy. The middle one-third of each testis with 3-5 cut on capsule was fixed in Boins and processed for histology. Five microns thick section was stained with H&E and PAS reagent. Results: This study revealed sever abnormalities of spermatogenesis in SCI rats especially during 4 weeks after surgery, SCI, characterized by delay in spermiation, Phagocytosis of elongated spermatid (presence of polynucleated cell mass with pyknotic nuclei), incomplete spermatogenesis, disorganized and incomplete cellular association of seminiferous epithelium, vacolization of epithelium and presence of polynucleated cell mass with pyknotic nuclei and signification decrease in the volume percent of seminiferous epithelium. Conclusion: This study demonstrates spermatogenic abnormality induced by SCI, that this may be indicated by neural cell death after SCI or hypothamo-hypophys-testis axis dysfunction.

Introduction: This study was started to improve in vitro blastocyst development and avoiding of blastocyst collapsment (no visibie cavity, returning of hatching blastocyst into zona pellucida, and tendency to degeneration). Materials and Methods: 24hrs one - cell mouse embryos were recovered from positive plug females and cultured in following canditions: Experiments 1,2: During first 48hrs NMRI 1-cell mouse embryos were cultured in R1 and then transferred to R2 (group 1) for 3 days. Some of one-cell mouse embryos were clutured in R1 plus commercially vero line or fibroblast cells derived from 12-14 days mouse embryos. After 48hrs, embryos were transferred to R2 plus one of the cell lines (groups 2, 3). Experiment 3, 4: One-cell mouse embryos were cultured in R1 for 48hrs and then transferred to R2 plus vero cells or R2 plus embryonic fibroblast cells, (group 5) for 3 days. Experiment 5: The 24hrs and 48hrs R2-conditioned mediums of vero and embryonic fibroblasts cells were prepared by filtration through 0.22 µ filters and freezed in-20oC. 1-cell mouse embryose were cultured in R1 for 48hrs and transfeered to 24hrs or 48hrs-R1 conditioned mediums for other 3 days. Results: Comparsion of blastocyst development in 1st, 2nd and 3th group did not show any significant difference (59%, 54 and 52% respectively). Howerer, a high percent of 1-cell embryo developed as hatching and hatched blastocyst (71%) in both cocultures after 5days. The conditioned media (24hrs and 48hrs) on both type cell lines were not improved the blastocyst development and lot of blastocysts were collapsed in 5th day. Conclusion: These results showed that blastocyst development was significantly improved and blastocyst collapsment was markedly reduced, by using only sequential culture media during 2 days after one-cell embryo retrieval and simultaneously with coculture during following days. 2) Conditioned media did not improve blastocyst development.

Introduction: Vanadium salts have been suggested as a possible therapeutic agent for the treatment of diabetes. The aim of the present study was to clarify immunohistochemical changes that occur in the pancreatic beta ([Pancreatic Islet Beta]) cells of vanadyl sulphate (VS) treated STZ induced diabetic rats. Materials and Methods: Male wistar rats were made diabetic with injecting a single intravenous dose (40 mg/kg) of STZ and were divided into two groups. In the first group (DT) VS was administered in the drinking water at a concentration of 1 mg/ml and treatment was maintained until normoglycemia appeared. Second group of animals received distilled water for the same period as the DT rats end of the and was considered as control diabetic (DC) group. One group of animals (NC) was injected intravenously with the same amounth of veicle as the diabetic rats and was considered as non-diabetic control. In the end of the experiment all of the animals were killed and Horseradish Peroxidase procedure performed on pancreatic islet paraffin sections using guinea pig antiserum to insulin as primary antibody. Results: VS treatment was accompanied by amelioration of the signs of diabetes in DT rats while DC animals remained diabetic during the period of study. In immunohistochemistry while Insulin immunoreactive [Pancreatic Islet Beta] cells accounted for the majority of the islet cells in DT rats, immunostaining of pancreatic islets of DC rats revealed a considerable reduction in the number of Insulin immunoreactive [Pancreatic Islet Beta]cells. Conclusion: The results of this study indicate that the amelioration of the diabetic state in DT rats was accompanied with well preservation of pancreatic islets and insulin immunoreactivity in [Pancreatic Islet Beta] cells.

Introduction: The septal area is an important telencephalic center that plays a prominent role in the learning, emotional sexual behaviour processes and autonomic functions. This area contains two main divisions; lateral (LS) and medial (MS), that receive numerus afferents from hippocampal formation. This study compares, quantitatively distributional pattern of the afferent neurons to these two major subdivisions of septal area from hippocampal formation. Materials and Methods: In two different groups of male rats, stereotoxic injection of one microlitre HRP (25%) by using hamilton syringe into medial and lateral septal areas. After 48-72 hours, the brains were perfused and fixed through the left ventricle. The brains were removed and 70 micrometer coronal sections were cut from the basal forebrain part. TMB-histochemical reaction was performed on them. Then sections were counterstained by Neutral Red. Results: Following injection into each part, retrogradely labeled neurons were observed in the CA1-2 fields of Ammons horn, presubiculum and subiculum. Number of labeled neurons following injection into lateral septum were highly more than that of injection into the medial septum. In comparison to MS group, the neurons in LS group were also more intensely stained. Conclusion: Regarding to the results of this study it could be concluded that however, both lateral and medial septum receive afferents from hippocampal formation, there is a strong quantitative distributional discrepancy on the hippocampal origin of these afferents, that could be the basic for different functional roles of these areas.