Cell Journal (Yakhteh)
Year:2008 | Volume:9 | Issue:4 (36)|Papers Count:9

Objective: The aim of this study was to examine the efficiency of HA-binding test and sperm gradient processing method for the selection of sperm with normal protamine, intact DNA and normal morphology.Materials and Methods: The study was conducted on 29 patients undergoing ICSI. Semen analysis was carried out according to WHO criteria. Semen samples were divided into three equal portions. One portion was left untreated (control), the second portion was used for HA-binding (HA: Hyaluronic acid) and the third portion underwent PureSperm gradient processing. Then, the three portions were evaluated to examine protamine deficiency (Chromomycin A3 staining), sperm morphol­ogy (Papanicolaou staining) and DNA integrity (Sperm Chromatin Dispersion Test). Coefficients of correlation and student t-test were carried out using SPSS 11.5 and P-value lower than 0.05 was considered as significant. Results: The mean number of sperm abnormality, protamin deficiency, and DNA fragmentation in HA binding and sperm gradient processing groups were significantly decreased as compared to the control group (p<0.05). In addition, sperm gradient processing was superior to HA-binding method in the selection of sperm with normal protamine, intact DNA and normal morphology.Conclusion: Both HA-binding and sperm gradient processing methods were effective in the selection of sperm with better quality in terms of normal morphology, normal protamine content and DNA integrity. However, sperm gradient processing was superior to HA-binding method in the selection of normal sperm.

Objective: The purpose of this study was to investigate the role of fast inactivating (IA) voltage dependent potassium channels in shaping the discharge activity of cerebellar Purkinje cells (PCs) in ataxic rats.Materials and Methods: Male Sprauge-Dawley rats (40-60g) were used in this study. Ataxia was induced via single i.p. injection of 3-acethylpyridine (65mg/kg). Four days following induction of ataxia, the rats were decapitated after being anesthetized using ether. 300mm thick parasagittal slices were prepared from the vermis of the cerebellum. Sharp intracellular recording was achieved from Purkinje cells under continuous superfusion of ACSF containing synaptic blockers.Results: Intracellular recording of 21 Purkinje neurons in cerebellar slice prepara­tion showed that Purkinje cells of ataxic rats fire in three modes of tonic (23.81%), burst (19.05%) and silence (57.14%). Following the application of 4-AP, 83.33% of the silent PCs were spontaneously fired and the burst activity was robustly enhanced in bursting PCs. In addition, 4-AP significantly increased the duration of burst and active periods but decreased the duration of inactive period. It also increased the number of sodium spikes within each burst.Conclusion: 4-AP sensitive current in Purkinje cell of ataxic rat involves in three modes of firing pattern. In addition, the inhibition of fast inactivating potassium channels (IA) with 4-AP enhances the spontaneous discharge activity of Purkinje cells.

Objective: This study was carried out to examine the effect of vero cells co-culture on developmental competence of immature oocytes. Materials and Methods: Bovine cumulus-oocyte-complexes (COCs) were matured in either presence or absence of vero cells. Matured oocytes were inseminated and cultured for up to nine days. Cleavage percentages were recorded on day two post insemination (pi). Embryos were evaluated on daily basis till 9pi. Expanding/expanded and hatching/hatched blastocysts were used for cell number assay. Results: The results indicated a significantl increase in the cleavage number in oocytes matured in the presence of vero cells than that of the control (86% vs. 76%). The percentages of advanced embryos appear to be greater on a daily basis in COCs matured in the presence of vero cells compared to those of the control. However, these differences were not significant. Blastocysts derived from COCs matured in the presence of vero cells had a significantly higher number of inner cell mass, trophectoderm and total cell number in expanding/expand and hatching/hatched embryos in comparison to those of the control. Conclusion: Results confirm that co-culture of bovine COCs, during in vitro matura­tion, enhances the potential for cleavage and for producing blastocysts with higher quality.

Objective: The current study was undertaken to examine the bactericide effects of atmospheric plasma produced by flat and comb electrodes on E.coli.Materials and Methods: DC (20 W) and AC (500 W) power supplies, flat and comb electrodes, insulator and oxygen gas were used to produce atmospheric plasma. Bactericide effects of the atmospheric plasma were studied on E.coli on sterilized plate placed on ice powder, at 3 McFarland concentration.Results: Flow of the atmospheric plasma produced by flat electrodes and AC power supply reduced E.coli population by 100% during 15 minutes. But the flow of the atmospheric plasma produced by comb electrodes reduced E.coli population by 12% in 15 minutes and 51% in 30 minutes. In addition, the flow of atmospheric plasma produced with DC power supply with flat and comb electrodes reduced E.coli population about 50% and 17%, respectively in 30 minutes. Statistical analysis indicated significant differences (p<0.005) between bactericide effects of atmospheric plasma from DC and AC power supply in 15 and 30 minutes.Conclusion: Flat electrode structures in both down and high plasma atmospheric sterilize and control E.coli populations. We recommend further studies on examining the bactericide properties of atmospheric plasma on other bacteria, especially staphylococcus aureus and bacillus subtilis.

Objective: Purple cone flower plant (Echinacea purpurea) is one of the mostimportant Herbal products in many countries. Up to now a lot of experiments demonstrated the controversial effects of this herb on immune system. In this research we study the in vivo and in vitro effect of Iranian E.purpurea extract on cellular immunity.Materials and Methods: At first we determined the lethal dose of E.purpurea extract after intraperitoneal injection in BALB/c mice. Then we made five groups of mice and treat them by four times intraperitoneal injection of 1 ml extract at different doses (0, 0.4, 2, 10 and 50mg/ml) during two weeks. Splenocyte proliferation response to extract was assessed by MTT method. Delayed-Type Hypersensitivity (DTH) response was evaluated by priming mice with 1×108 Sheep Red Blood Cell (SRBC) injected subcutaneously in the back on day 7after treatment. Results: As a result no significant variation in weight and spleen index of test groups to control was observed. Splenocyte proliferation and DTH response of test groups to control increased significantly (p<0.05).Conclusion: However these data confirmed the results of previous studies, inaddition presented the first results about significant increase in DTH response that could not be seen before. Scince the main reason of this difference refers to active compounds of herb extract, comparing effective component of this extract with that of E.purpurea cultivated in other geographical condition will consider as the next studies.

Objective: In this investigation Murine Embryonic Stem (ES) cells were differentiated into endothelial cells.Materials and Methods: Murine ES cells (CCE cell line) exposed to Alpha-MEM medium containing 10% FBS for 4 days. Then obtained Flk-1 (Flk-1: Vascular Endothelial Growth Factor Receptor 2) Positive cells were cultuted in Endothelial Growth Medium-2 (EGM-2) until the last day of experiment. Differentiated cells were evaluated by immunocytochemistry, RT-PCR and Tube Formation Assays.Results: When the ES cells cultured in collagen coated dishes containing Alpha-MEM & FBS, Flk-1 positive cells were obtained. After transfering Flk-1 positive cells into fibronectin coated dishes containing EGM2, the cells were assumed a relatively uniform endothelial cell morphology and could be propagated and expanded. Immunocytochemical and RT-PCR analysis of differentiated cells showed that they take up acetylated low-density lipoprotein (LDL), express Flk-1, CD31 and bind the BS-I lectin. When placed in Matrigel, these Murine ES cell–derived endothelial cells formed capillary-like structures characteristic of endothelial cells.Conclusion: ES cell–derived endothelial cells provide a novel means to examine the mechanisms of endothelial cell development, and may open up new therapeutic strategies.

Objective: Strong evidence indicate that activity of glucocorticoid receptors is necessary for emotional memory processess and these effects probably are mediated by hipppocampal opioid receptors. The aim of this study was to test whether there were any interaction between glucocorticoid and hippocampal opiate receptors on consolidtaion and retrieval of spatial memory. Materials and Methods: In this experimental study, 120 male Wistar rats (250-300 gr) were used. At first, the animals underwent surgery implant canulae in bilateraly of dorsal hippocampus (DH). Then one week later they were trained in a Water Maze task with eight trials for one day for consolidation and six trials per day for six consecutive days for retrieval processes. The retention of the spatial training was assessed 24h after the last training session with a 60-s probe trial. Corticosterone (1mg/kg the best dose) was injected IP immediately after training or 30 min before retrieval. Also Nalthroxone (10 and 20mg/ul/ per side) was injected bilaterally into hippocmapus immediately after training or 60 min before retrieval test.Results: The results indicated that injection of corticostrone immediatly after training or 30 min before retention significantly enhanced or impaired consolidation and retrieval memory respectively (p<0.01) and these effects were blocked by injection of naltroxone in to DH (p<0.05) in either cases.Conclusion: The findings suggest that the opioid receptors in hippocampus play an important role in mediating the enhancing or impairing effects of systemic corticosterone on memory consolidation and retrieval.

Oxidative stress arises when there is a marked imbalance between the production and removal of reactive oxygen species that may lead to several diseases. Cadmium is an environmental and industrial pollutant that results in formation of free radicals. The aim of this study was to determine the oxidative stress status in cad­mium exposure workers.This is a cross-sectional study of 36 people of cadmium exposure workers with two years employment records who were randomly selected as the case group and 36 other people who were selected with matched age and gender as the control group. Both groups were compared for the different parameters of oxidative stress.The mean and standard deviation (mean SD) of the total antioxidant capacity of serum was 2.9±0.9 in case group and 2.4±1.2 in control group (p=0.003). The rate of lipid peroxidation was 7.2±3.4 in case group and 9.3±3.6 in control group (p=0.015). The rate of total thiol group was 34±243 in case group and 0.246±0.114 in control group (p=0.004).The results of the study indicated that following exposure to cadmium, their TAC and thiol group antioxidants markedly enhanced to retaliate the influence of excessive reactive oxygen radicals. This decreased lipid peroxidation.