Cell Journal (Yakhteh)
Year:2003 | Volume:5 | Issue:18|Papers Count:9

Introduction: This study was performed to investigate the effect of vitrification procedure using ethyleneglycol as cryoprotectant on the follicular morphology of neonatal ovarian tissue of mouse after thawing andautotransplantation.   Material and Methods: Ovaries from 3 weeks neonate NMRI mice were excised then vitrified in asolution of DMEM medium containing ficol 70, sucrose, acetanied and ethylene glycol (EGFS 40%) andstored in liquid nitrogen. After thawing with 1 M sucrose solution and equilibration with DMEM medium, some vitrified-warmed andfresh ovarian tissues were studied morphologically and the others were autografted intraperitoneally. Fourweeks after transplantation, animals were sacrificed and the fresh and grafted samples were fixed and studiedunder light microscopy.   Results: Our results showed that there was no statistically significant difference between the normalfollicles in vitrified, toxicity tested and fresh control ovaries. After transplantation statistically significant differences was observed between normal follicles in intactcontrol, transplanted-control and vitrified-transplanted groups (P<0.014) and these rates were 98.18% ,89.33% and 91.54% respectively. In the transplanted group the number of follicles had a relative decrease andthe ovarian tissue was invaded by fibrous and fat tissue. However follicles in different developmental stageswere located in the periphery of ovarian tissue.   Conclusion: Our observations showed that this procedure is useful and efficient for preservation ofneonatal mouse ovarian tissues and can be used for preserving the reproductive potential of infertile patientsin the future. However it is necessary to improve maturation conditions such as changing the site oftransplantation.        

Introduction: Different environmental factors such as drugs, ionizing radiation, etc can induce apoptosis.In the present study the effects of dose- rate and post-irradiation incubation time on gamma radiation inducedapoptosis in human breast carcinoma cell line (T-47D) (Invasive ductal carcinoma) was investigated. Material and Methods: The T-47D cells were cultured in T25 flasks and during log phase of growth,the cells were counted and exposed to 4,8,12 and 16Gy gamma radiation. Untreated and etoposide treated cellsserved as negative and positive controls, respectively. At 24,48 and 72 hours post irradiation, the cells wereharvested and stained with HO/PI and AO/PI for morphological and flow cytometric analysis, respectively.Absolute and relative numbers of live(L), apoptotic (Ap) and dead (D) cells were determined using the abovementioned methods and reported as Mean ± SD. Results: In comparison to normal controls, gamma radiation inhibited growth of T-47D cells in a dose andpost-irradiation incubation time dependent manner which was defined by reduction of cell proliferation justafter exposure that continued for 48 hours and was followed by cell count reduction. The relation between irradiation dose and apoptosis induction was not linear and inverted at 8Gy after 48 hours post irradiation.Therefore, maximum apoptotic response (%) occured during 48 hours after expossure to 8Gy. However,relative numbers of a live and dead cells decreased and increased respectively in a dose and time dependentmanner. Conclusions: The present study revealed that gamma-irradiation could induce apoptosis in T-47D cellline and the rate of apoptosis was radiation dose dependent but this relation was not linear and in higherdoses became inverted . However, cell viability and death were dose and time dependent.        

Introduction: Obstructive cholestasis is associated with overproduction of endogenous opioids (EOP) ,nitric oxide (NO) and cytokines in the blood stream. These consequences can affect sex hormones. Sincefertility is a result of physiological balance of sex hormones, we investigated the relationship betweenobstructive cholestasis and sex hormones and apoptotic germ cells in adult male rats. Material and Methods: To study this, we used three groups of animals: Control (No-surgery), Sham(surgical control), and cholestatic (surgical ligation of the bile duct). After 3 weeks all animal were killed byether, and serum concentrations of inhibin B , FSH and LH were determined by ELISA andRadioimmunoassay respectively. Testicular germinal cell apoptosis was evaluated by DNA fragmentationdetected by in situ terminal deoxynucloetidyl Transfrase-mediated dUTP nike end labeling (TUNEL). Results: The findings of this study showed that LH and FSH levels were significantly decreased incholestatic compared to control and sham groups (P<0.05). However, inhibin B level was significantly higherin cholestasis compared to control and sham (P<0.05). On the other hand, no statistically significant differencewas seen between germinal cell apoptotic index of cholestatic group and that of the other groups (P>0.19). Conclusion: These findings revealed that although the serum level change has no significant effect ontesticular germinal cell apoptosis. We speculate that testicular germinal cell apoptosis is not just dependent ongonadoteropin hormone and that other factors may be involved.        

Introduction: Determination of the efficacy of crude Entamoeba histolytica extract and its fractions (FI,FII & FIII) in antiamoebic antibody detection by ELISA. Material and Methods: Entamoeba histolytica NIH:200 was cultured axenically and different fractionsof its crude amoebic extract, i.e. FI, FII & FIII, were obtained by column chromatography. Efficacy of Crudeand fractioned antigens for antiamoebic detection by ELISA was compared in acute amoebiasis cases. For thispurpose, a total of 25 cases including 15 amoebic liver abscess cases, 10 acute amoebic dysentery and 10control cases were included into the study. Results: Crude Entamoeba histolytica extract and fraction I are more useful for antiamoebic antibodydetection. While, efficacy of FII & FIII is lower and theses two extracts are not as effective in antiamoebicantibody detection. By using these antigens the sensitivity of ELISA reduces while using FI for ELISA hasquite a high sensitivity. Conclusion: In intestinal and extraintestinal invasive amoebiasis humoral immune response isaccompanied by antibody protection against amoebic antigens. Antibody present in sera of study groups weredetected by employing ELISA and it was found that ELISA, especially by using fraction I of Entamoeba histolytica extract, is 90_100% specific in detecting antiamoebic antibody.    

Introduction: This study was performed to analyze chromosomal aletrations in G1 and G2 immortalized MOLT-4 cell line following treatment with methotrexate (MTX) and cytarabine (ara-C) alone or incombination.   Material and Methods: MOLT-4 cells were cultured and manitained in RPMI-1640 supplementedwith fetal calf serum, antibiotics and L-gluthamine. Cells were treated with ara-C and MTX with a concentration of 100µmol/L at G1 and 50µmol/L at G2. One and half hour prior to harvesting, cell wereexposed to colcemid. (0.1µg/ml find concentration). Microscopic slides were prepared using standard1000).‚procedure. After staining with 5% Giemsa, metaphase was evaluated using light microscope (×1000).   Results: Results indicate that ara-C and MTX, at a 100µmol/L concentration, used for treatment of G1cells, increase the frequency of chromosomal type aberrations; although ara-C was more potent in aberrationinduction. The effect of combined treatment with ara-C and MTX on G1 cells was more than effect of eachdrug when used alone. The effect of these drugs on G2 cells with a concentration of 50µmol/L was found to besignificantly different with that of controls (P<0.05). The frequency of chromatid type breaks induced by ara-C was considerably higher than MTX. When these drug were uaed in combination , the frequencuy of aberration increased dramatically.   Conclusion: Results indicate that both ara-C and MTX produce clastogenic effects on MOLT-4 cells.Combination treatment with ara-C and MTX has an additive effect on G1-cells. But a synergistic effect wasfound when they were used for treatment of G2-cells. The reason for hypersensitivity of G2-cells to chemicalsmight be due to shorter repair time of lesions induced in DNA, which are experessed as chromatid typeaberrations in the first mitosis.                                    

Introduction: The aim of this study was to determine the orphological and ultrastructural changes ofampullary portion of mouse fallopian tube after ovarian hyperstimulation and progesterone injection duringimplantation period. Material and Methods: For this purpose 6-10 weeks old NMRI mice were hyperstimulated using hCGand hMG injections. In one group of hyperstimulated mice daily injection of progesterone was performed.Non-stimulated control groups and stimulated groups were pseudopregnanted artificially. Three to four daysafter hCG injection, the mice were sacrificed by cervical dislocation. The samples, which were obtained fromfallopin tube were processed for light (H&E and PAS stainning) and electron microscopic studies. Results: There were no noticeable morphological and ultrastructural changes between the hyperstimulatedand control groups. It seems that the number of non-ciliated cells were increased on the fourth day ofpregnancy and or hCG injection. The cytoplasm of these cells were denser than the ciliated cells. Weak PASreaction was seen on the surface epithelium but this reaction was stronger in the lamina propria. Conclusion: Hyperstimulation could not alter the morphology and ultrastructure of ampullary portion of mouse epithelium during implantation periods.    

Introduction: During the recent years many studies have been performed on the effects of nitric oxide andoxygen-derived free radicals (OFR) on the aorta, but there is no report on the effects of simultaneousmanipulation of both systems on the relaxation of this artery. Material and Methods: After anaesthesia, aortic rings from male rats (320-420 g) were obtained. The pieces of aorta were mounted in organ bath and attached to physiograph. The study groups consisted of SNP (n=6,10-10-10-5M ) and sodium nitroprusside) and SNP+DMTU groups (n=6 8×10-5M dimethyle thiourea, DMTU+SNP) Student t-test was used to compare the mean of IC50 of these groups. Results: This study showed that, administration of SNP induces relaxation of rat aorta and this effect isincreased significantly by simultaneous administration of DMTU (IC50=1/975 &10 -10 M, P<0.0003). Conclusion: We concluded that simultaneous administration of moderate dose of OFR scavengers andnitric oxide donors induces arterial relaxations, which may be useful in control of arterial flow and pressure inconditions such as ischemia-reperfusion.    

Introduction: In this study, the role of adenosine A1 receptors of hippocampal CA1 region neurons onentorhinal cortex kindled seizures was investigated.   Material and Methods: A tripolar(stimulating and recording) electrode was implanted in entorhinalcortex and two guide cannulae were implanted in the CA1 region of the left and right hippocampi of allanimals. A week after surgery, animals were stimulated daily to be kindled. In full kindled animals, N6-cyclohexyladenosine (CHA; 1, 10 and 50µM  a selective A1 receptor agonist ) and 1,3- dimethyl -8-cyclohexylxanthine (CPT; 5 and 10µM a selective A1 receptor antagonist) were microinfused (1µl/2min) intothe CA1  region of hippocampus and animals were stimulated at 5, 15 and 120 min after drug injection. Allanimals received artificial cerebrospinal fluid, 24 h before each drug injection and the results were used ascontrol.   Results: Obtained data showed that 5 and 15 min after injection, CHA at concentrations of 10 and 50 µM,reduced entorhinal cortex afterdischarge and stage 5 seizure durations and increased stage 4 latency. It alsodecreased seizure duration only at concentration  of 50µM. CHA (1 µM) did not alter seizure parameters. CPTat concentration of 5µM could not produce any changes in seizure parameters, but at concentration of 10 µM increased after discharge and stage 5 durations and decreased stage 4 latency. Intrahippocampal (CA1 region)pretreatment with CPT (5 µM) before CHA (10 and 50 µM) reduced the effects of  CHA on seizureparameters .    Conclusion: It may be suggested that hippocampal CA1 region plays an important role in seizurepropagation from entorhinal cortex to other brain region/s and activation of  adenosine A1 receptors in thisregion have anticonvulsant effects on entorhinal cortex kindled seizures     .