Cell Journal (Yakhteh)
Year:2001 | Volume:3 | Issue:9|Papers Count:8

Introduction: Sperm Viability Test is important in diagnosis and treatment of male infertility. The common test is E&N and HOST which they are based on membrane permeability and integrity. The aim of this study was to evaluate MTT Assay as a sperm viability test based on mitochondrial activity. Materials and Methods: Washed sperm samples were co-incubated with MTT in different media and different times in order to obtain the best condition for carrying out MTT assay. Then coefficient of variation of MTT was obtained and sensitivity and specificity of each test were calculated. Then MTT and E&N and HOST were carried out on 57 samples from infertile patient referring into Isfahan Fertility and Infertility Center. Then correlation coefficient of these tests with each other and sperm motility were obtained using SPSS software statistical program.Results: Ham's F-10 + 15 mM Hepes + 10% HSA at pH=7.4 at 37° c for 2 hours were the best condition for carrying out Sperm MTT Viability Assay in order to obtain optimal results. MTT assay showed a good significant correlation with HOST and E&N and sperm motility. All the test had high sensitivity but the specificity of HOST was less than that of E&N and MTT.Conclusion: MTT assay appears to be a suitable sperm viability test with high sensitivity and specificity and low coefficient of variation. This test designed not only for differentiating but also selecting and sperating viable sperm from dead sperm. Therefore this test may have an application for intra-cytoplasmic immotile sperm injection which remains to be study.

Introduction: This study was performed to investigate the effect of vitrification procedure using ethylene glycol as cryoprotectant on the ultrastructure and morphology of primary follicles of mouse ovarian tissue. Materials and methods: Ovaries from 8-10 weeks old NMRI mice were vitrified using a solution of T6 medium containing 30% (W/V) Ficol 70, 0.5 M Sucrose, 10.7% (V/V) Acetamide and 40% (V/V) Ethylene glycol (EGFS 40) and stored in liquid nitrogen. After thawing the vitrified ovarian tissues were washed by 1 M sucrose solution and equilibrated by T6 medium. For histology, the frozen-thewed and fresh ovarian tissues were fixed using Bouin"s solution. The fixed tissues were embedded in paraffin wax, serially sectioned at 6 micrometer and stained with Hamatoxylin and Eosin. For electron microscopic studies, the frozen-thawed ovarian and control tissues were fixed with 2.5% Glutaraldehyde and 1% Osmiume tetroxide. After dehydration with ethanol the tissue were embedded in Epon 812 resin. Ultrathin sections were stained with Uranyl acetate and Lead citrate, then examined under TEM.Results: The proportion of morphologically normal follicles was observered frozen and fresh ovaries. At the light microscopic level almost all of the primary follicles appeared normal in frozen and non-frozen ovaries.In electron microscopic study the integrity of cell organelles, nuleus of oocyte and follicular cells were well preserved, but some mitochondria were swell, their cristae disappeared in some parts, perivitilline space was electron lucent and there was few cortical granule in cortex of oocyte.Conclusion: The results of this study showed that the vitrification method, using ethylene glycol as cryoprotectant did not cause any harmful damage to follicular cells and oocytes of primary follicles, thus this procedure could be an efficient method for storage of immature oocytes within ovary especially before cancer therapy.

Introduction: Retinoic acid (RA), one of the syntetic vitamin A derivatives which has been frequently used as a drug to treat face acne, is known to have specific side effects on the testis, liver, and spleen, it is also known as an oxidative agent in high doses. Therefore, we have tried to find a compound which would decrease its side effects. Curcumin which is found in the rhizome of turmeric (Curcuma longa) is known to have antioxidant, antiinflamation and antibacterial properties. It has also shown that this chemical can decrease lipid proxidation and scavenge free radicals. In order to investigate possible protective interaction between curcumin as a strong antioxidant with side effects of RA as an oxidant agent, we conduct a series of experiments involving dietary application of RA and curcumin with white mice. Materials and Methods: In this research we have chosen 6 weeks old male mice of NMRI strain weighting 262 grams. The mice were divided into five experimental groups. The first group were fed only with normal diet (as Control) 1, but the other four groups were fed with normal diet plus olive oil (as Solvent) 2, (Retinoic acid) 3, (Curcumin) 4 and (Curcumin + Retinoic acid) 5. We administrated 10 mg/kg of curcumin and 30 mg/kg of retinoic acid for eight days by gavage method. Following dissection of mice and collection blood various parameters including cholesterol, triglycerides, LDH and ALP were analyzed with laboratory kits. The analysis of the variance and Tukey's test were used to compare the statistical difference among the parameters.Results: Results obtained from these experiment indicated that RA increased the amount of cholesterol and triglyceride by 23.3% (P<0.001) and 3.11% (P<0.05) respectively compared with the control group. Similarly, the amount ot LDH and ALP activity enzymes occuring in blood plasma increased by 25.8% and 42.4% (P<0.001) respectively compared with solvent group (Control). Dietary uptake of curcumin has decreased the amount of cholesterol and triglyceride by 12.2% (P<0.01) and 25.2% (P<0.01) respectively. Mice adminstered with dietary curcumin have demonstrated a marked decreased in LDH and ALP activities by 11.2% and 15.7% (P<0.05) respectively.Conclusion: It was obsereved that the cytotoxicity of retinoic acid had been decreased markedly by curcumin. Therefore anatioxidant role of curcumin can decrease side effects of retinoic acid. It can be concluded that curcumin can be used for treatment against RA side effects.

Introduction: It is clear that the phenol can absorb through skin and mucosal surfaces, and affect on different systems of body and ultimatly cause death. Furthermore during the pregnancy, phenol may pass through the placental barrier and therefore enter the embryonic circulation and effect on organogenesis. The aim of this investigation was to study the embryonic effects of phenol in the devloping mice embryo. Materials and Methods: In this study pregnant mice (experimental group) received 100 mg/kg phenol via interaperitoneally injection during 8, 9 and 10 postmating days. The same manner control group was received physiological serum. At the last day of pregnancy, all of cases (experimental and control) were anesthetized, sacrificed and their fetuses collected and examined. In addition their weight and Crown Rump (CR) measured.Results: The result show a decrease in number of fetuses, high incidence of growth retardation in (P<0.05) and decrease of CR (P<0.005) in experimental group. Furthermore, absorption and mortality among fetuses experimental group as compared with control group (P<0.001).Conclusion: This study suggested that the phenol crosses placenta and able to produce embryotoxicity and embryolethality in experimental group.

Introduction: This study was performed to establish a complementary method to the diagnosis of complete hydatidiform mole and its differentiation from the other cases of gestational trophoblastic disease by measuring the total (ALP) and placental alkaline phosphatase (PLAP) specific activity. Materials and Methods: Serum and tissue extracts from 13 patients, 13 normal non-pregnant and 30 pregnant females were compared for these enzymes activities. Paranitrophenyl phosphate was used as substrate. Placental alkaline phosphatase was simply distingushed from its isoenzymes by its heat stability (65°c for 15min).Results: According to our findings from serum and tissue extract examination the activity of total alkaline phosphatase and placental alkaline phosphatase activities were reduced in hydatidiform mole patients in comparsion with pregnant females (P<0.05) but there was no significant difference in non - pregnant subject. Conclusion: These results show a severe hypophosphatasia in serum and tissue extracts of the patients. The reduction in activity of enzymes may be caused by low synthesis of enzyme by trophoblast cells or some post transcriptional changes that causse the reduction of enzyme activity. Since the ALP is probably important in active transport of suger and phosphate across the trophoblast cell membreanes, so the hydropic villi of hydatidiform mole placenta may due to reduction of enzyme activity in placental villi. Our results suggest that measurment of ALP and PLAP activity in serum and tissue extract can be a complementary method in diagnosis of hydatidiform mole.

Introduction: To recognize the myelinated nerve fibers of the external capsule in human brain. Materials and Methods: 10 adult and normal brains (20 hemispheres) from both sexes were studied using 15 mm serial sections in all three cardinal planes after fixation and processing. These sections were stained by Klüver - Barrera and Heidenhain - Woelcke methods.Results: Some fibers from different parts of the cortex through corona radiata entered the dorsal border of the external capsule. These fibers moved ventraly and ventrocaudally toward the ventral border of the external capsule, and most of them entered the cerbral peduncles trans-, sub-, or retrolenticularly. No connection was seen between these fibers and subthalamic nucleus or substantia nigra. Some fibers passed through dorsal part of putamen and connected the external capsule with posterior limb of the internal capsule; Some fibers exchanged between the putamen and the external capsule, some of them connected the putamen with dorsal parts of the external capsule and the other fibers connected the putamen with ventral parts of the external capsule. Some fibers passed between rostrum of corpus callosum and ventral part of the external capsule. Dorsal to these fibers, there were some fibers which connected the external capsule with ventral part of accumbens nucleus. Some fibers traced from the external capsule to posterior bundle of the anterior commissure; some of them entered the commissure but others terminated in nucleus basalis (Meynert) neurons. Some fibers passed through rostral part of the external capsule and connected the frontal lobe with medial dorsal nucleus of thalamus.Conclusion: The external capsule is a part of the white matter of the cerebral hemisphere which contains different variety of nerve bundles. The main component of the external capsule is the corticotegmental fibers, especialy in dorsal parts. This capsule provides very important pathways for connecting the cortex to subcortical nuclei as well as the different cortical areas with each others.

Introduction: During kidney and other organ's transplantation, the organ to be transplanted, must inevitably remain out of the body with little or no blood perfusion at all, for long period of time (Ischemia). This causes the release of oxygen free radicals (OFR). Reperfusion (reintroduction of blood flow) will further exacerbate the initial damage caused by the ischemic insult and may result in the production of free radicals.Objective: The goal of this study was to investigate the effect of ischemia- reperfusion (IR)(30 min ischemia followed by 10 min reperfusion)by clamping and decamping of the renal artery on the oxidant status of renal tissue and venous plasma.Materials and Methods: White male rats with average weight of 26040 grams were randomly divided into sham-operated, control and ischemia-reperfusion (30 min ischemia, followed by 10 min reperfusion) groups (n= 7 each group). In the control group, 70 min after dissecting the abdomen, renal blood and tissue were removed. In the IR group, 30 min after dissecting the abdomen, 30 min ischemia (clamping of renal artery) and 10 min reperfusion (declamping of renal artery) were applied, then, the kidney and blood were sampled. Vitamin E (as an endogenous antioxidant) was extracted from the right kidney tissue and the venous plasma and measured by HPLC.Results: The amount of vitamin E (َmol/L) in plasma of IR group had a significant decrease in respect to the control group (P<0.002). The same was true for kidney tissue (P<0.0004).Conclusion: This study suggests that during reperfusion, the body uses all its defence mechanism to fight against the produced OFR the measured vitamin E showed the oxidative state of an organ (in this case kidney) wich would be of benefit.

Introduction: Kidney is one of the target organs in diabetes mellitus. Diabetic nephropathy is a major cause of end stage-renal disease and resulting an increased in morbidity and mortality of effected individuals. Since diabetes is a progressive disease, therefore early diagnosis leads to better treatment. In this study primary diagnostic parameters were the goal.Materials and Methods: In this study morphometrical and stereological methods and stains of H&E and PAS were applied. Experimental rats were classified in to two groups, 8 and 17 weeks after treatment, the animals with alloxan.Results: 17 weeks diabetic rats showed increased in glomerular measangium and glomerular volume. Conclusion: Therefor this study showed that glomerular changes happen in earlier time can be an early diagnosis method in affected kidney.