Anatomical Sciences Journal
Year:2006 | Volume:4 | Issue:1|Papers Count:8

Introduction: The aim of this study was to compare ultra structure of embryos obtained from young and old mouse oocytes.Materials and Methods: Metaphase II oocytes were retrieved from old NMRI mouse (7-9 months old) and young NMRI (10-12 weeks old) after ovarian stimulation. NMRI mouse sperms (10-12 weeks old) were added to T6 medium containing retrieved oocytes.After PN formation, embryos were transferred to culture medium (comtaining 15mg/ml BSA).Developmenting of embryos was recorded every 24 hours. 2 cell embryos after were fined in 2.5% glutardehid phosphate the samples dehydrated and embedded. Blokcs were sliced after timming. Seim thin (500 nm) and ultra thin (70 nm) sections had been (collected on copper grids) stained. Samples were studied by election microscopy.Results: At the (LM) level, embryos driven from young and old oocytes were surrounded by zona pellocida polar bodies and cellular fragmentation were seen in perivitelline space which was morefrequent in older embryos.In embryos driven from old oocytes, perivittelline space was more enpanded and microvillies were arragned reqularly. Nuclei of blastocytes were enchromatin in both embroyos. In old embryos, intramuscular anulatellae lamllae was ween which was secndary to folding of nucleous inhere membra.Mithochondies and intermediate filaments were seen in both embryos in cytoplasm. Which were similar in morphology and ultra structure. But their arrangement and distribution were different.In embryos dreven from oocytes. Most of mitochonderies were central and around the nucleuses.While in young embryos, they were distributed in cytoplasm.SER collection was higher in young embryos while residual bodies and lipid were more frequent in old embryos.Conclusion: This study revels that embryos driven from old oocytes had several ultra structure changes in comparison to young embryos. This may be one of the reason follow implantation and birth rate in old subjects.

Human embryonic stem cells (hESCs) represent a population of undifferentiated cells with both self renewal and multilineage differentiation characteristics representing all three embryonic germ layes thus revealing their pluriptent potential. Human ESCs originate from chromosomally euploid, aneuploid, and mutant human embryos that are available from in vitro fertilization clinics treating patients for infertility or preimplantation genetic diagnosis. The pluripoent nature of hESCs has attracted great interest in using them as an important source for functional genomics, drug screening, and, perhaps eventually, cell and gene therapy. The derivation of the first hESCs was reported in 1998. Since then we have learnt a great deal about how to isolate and culture these cells to use them for the aforementioned objectives. This review gives an verview of the different methods of establishment, characterization, and maintenance of hESCs.

Introduction: To evaluate the 8 layer discontinues puresperm gradient in separation of human spermatozoa according to sex chromosomes by fluorescent in situ hybridization (FISH). Material and Methods: This study was carried out on three patient referring to Royan fertility and infertility center. Semen analysis was assessed according to WHO criteria. Each sample was divided into two aliquots, one as a control was washed with phosphate bufer saline and another one for processing through 8- step discontinues puresperm gradient 35%-84% (7% differences between each subsequent layers). After centrifugation, sperm from top and bottom fraction was aspirated and fixed on slide, FISH procedure was performad by using X and Y specific probes. DNA probes bind to alpha satellite complementary sequence on centromeric region of X and Y chromosomes. More than 1000 sperm were scored for each sample. The proportion of X and Y bearing sperm chromosomes was determined by presence of red or green florescent signals. Results: Statistical analysis by chi-square test revealed no significant differences in the X:Y probration in the 84% layer compared to the controls. But there were significantly differences in the 35% layer compared to 84% layers and the controls (P=0.001, P=0.006 respectively), and X bearing spermatozoa was enriched in bottom layer.Conclusion: The result of this study showed that 8-step discontinues Puresperm gradient is not so reliable for separating X and Y bearings spermatozoa.

Introduction: Leukemia inhibitory factor is a secreted glycoprotein with a range of molecular weight forms, from 38-67kd, resulting from differential glycosylation of protein of approximately 20Kd. LIF has the four a helix cytokine structure that contains six cysteine residues, already known to occur in many hematopoietic factor. LIF is expressed at high constitutive in human fallopian tubal epithelium. Some studies has been done on the efect of LIF on normal sperm motility and shown that this factor could improve normal sperm motility. Objective: In this project y knowing that there is no report avout the effect of LIF on low motility sperms of asthenospermic men we decide to determine effect of this factor on low motility sperms. Materials and Methods: In this investigation the sperm sample of 15 infertile men who have already referred to IVF department of Imam Khomeini hospital were collected and then preparing them by successive stages for culturing in Ham's F10+FCS 10% medium that contained various concentrations (0 ng/ml, 3 ng/ml, 5 ng/ml, 10 ng/ml, 50 ng/ml) of LIF at 37oC and under 5% of Co2 in air for up to 48 hours. Sperm motion characteristics were measured using Makler chamber. Sperm survival was determined by hyposmotic swelling test. Collected data with ANOVA method by means of SPSS software were analyzed. Results: Sperm Motility and survival were not significantly enhance until 6 hours exposure to different concentration of LIF but sperm progressive. Motility were significantly higher after 24h exposure to 10 ng/ml of LIF. P<0.05. The survival rate of sperm was significantly prolonged in that time exposure to 10 ng/ml of LIF P<0.05. progressive motility of sperms and survival rate were signiicantly higher after 48h exposure to 50 ng/ml of LIF. P<0.05. Conclusion: It can be concluded that the effect of Leukemia Inhibitory Factor on sperm motility and survival were concentration dependent and LIF increased sperm motility and survival in vitro after at least 24 hours of hours of incubation and we could not find significantly differences in shorter incubation period.

Introduction: Previous studies have shown that thyroid hormones (T3 and T4) play importand role for postnatal development and maturation. In this study, the effect of hypothyroidism on spermatogenesis 2 month age Wistar rats were investigated.Materials and Methods: The animals were divided into experimental and control groups. The experimental group were made hypothyroid by chemical thyroidectomy with propylthyourasil (PTU). The control were remind untreated. After 3 week, the animals were scarified and testes were fixed in formalin for histological studies.Results: Our data indicated that, hypothyroidism resulted n significant reduction in seminiferous tubule and lumen diameter. Control ras showed active spermatogenesis whereas in hypothyroid rats, the proliferation and differentiation of germ cells were arrested and their number was decreased.Conclusion: The present study clearly indicates that hypothyroidism adversely affects spermatogenesis; it also indicates that thyroid hormones are essential for normal spermatogenesis.

Purpose: The effects of Amiodarone on lung tissue and morphologic changes of alveolar cells.Materials and Methods: in this study, we used 21 mature male rabbits and they were divided into control and two experimental groups. Daily 80 mg/kg amiodarone was injected intra peritoneally to the experimental animals for oe or two weeks. The control group received normal saline the same dose as amiodarone. Then the animals were anesthetized, perfused and the lung tissues were fixed in 2.5% glutaraldehyde and 4% paraformaldehyde. After the routine tissue processing was done, the specimens were blocked in paraffin and serial 8 mm sections was prepared. After staining with hematoxylin and eosin, the sections were studied by light microscope.Results: The acute changes in lung tissue such as congestion of alveolar capillaries, hyperplasia of pneumocytes type II, death and pyknosis of alveolar cells, thickening of alveolar membrane and existance of fibrin like materials in the cells was seen. Furthermore the average of the nucleus diameter was decreased but the alveolar membranes increased statistically (P<0.05).Conclusion: These results indicate that i.p administration of amiodarone to rabbits can cause severe damages in their lung tissue.

Sperm premature chromosomal condensaion (PCC) and failed oocyte activation are considered to result in failed fertilization post intracytoplasmic sperm injection (ICSI). Recent studies also suggested that sperm protamine deficiency may induce PCC. The ain of this study was to assess the effect of failed oocyte activation and protamine deficiency on failed ferilization post ICSI and to indicate whether these factors result fertilization failure, dependently or independently of each other.Materials and methods: This study consisted of 56 (first group) and 86 (second group) patients undergoing ICSI. After ICSI, the remaining processed samples were used for evaluation of protamine deficiency (Chromomycin A3 staining) and sperm morphology (Papanicolaou staining). 16-18 h post ICSI, oocytes were assessed for presence or absence of pronucleai in the both groups. In the second group te failed fertilized oocytes were chemically activated by ionomycin. Failed fertilized oocytes and chemically activated failed fertilized oocytes were fixed and stainde (Gimsa staining) for chromatin analysis.Reslts: Percentage of fertilization in the first and second groups of patients was 60.20% and 59.94%, respectively. In the second grop, percentage of fertilization rise to 83.72% following chemical activation. A significant negative correlation was observed between percentage of CMA3 positivity with ferilization rate in both groups of patients. A significant negative correlation was observed between percentage of PCC and fertilization rate in the first group while a significant posigive correlation was observed between percentage of PCC and CMA3 positivity in the second group.By subgrouping the patients into low and high CMA3 positivity in each group, the result reveal that ferilization rate, percentage of intact and PCC sperm were significantly different in the two subgroups in both groups of patients.Conclusion: the results of this study reveal that after failed oocyte activation, possibly due to sperm factors, protamine deficiency may induce failed fertilization by inducing PCC. However, mouse sperm lacking P2 protamine; have been shown to activate oocyte post ICSI, further emphasizaing these two factors may function independently of each ogher in failed fertilization.

Sirenomelia is a rare congenital anomaly that characterized by complete or incomplete fusion of lower lims and often associated with various malformations.In this report we describe one case of sirenomelia with multiple malformations including fibular agenesis, complete agenesis of anurectum, external genitalia and urinary tract, Inadditin, there was multiple anomaly upper limbs including agenosis of radius that is rarely seen in other reports.