Cell Journal (Yakhteh)
Year:2003 | Volume:4 | Issue:16|Papers Count:8

Introduction: Cryopreservation of single human spermatozoa is a new technique designed to improve the management of male infertility, it was first introduced in 1997 and was reported by others latter. The aim of this research was to evaluate the efficiency of this method in preserving either ejaculated or surgically retrieved spermatozoae. Materials and Methods: Spermatozoae were obtained from patients attending the ICSI program at Koassar Fertility Center, Tehran, Iran, and the cases were divided into three groups according to the method of sperm retrieval : a-ejaculated sperm, b-PESA and c-TESE (both with shaking sperm).For each group, 10 empty zonae were prepared using degenerated, immature or unfertilized human oocytes. Spermatozoa (8-10) were transferred inside each empty zona. The zonae were put in a droplet of follicular fluid + 15% glycerol (V/V) for five minutes before it was loaded in 0.25 ml straws and plunged in liquid nitrogen. After thawing, spermatozoa were extracted from the zonae and sperm viability and motility were recorded and analyzed statistically.Results: The results showed that less than 12% of the sperms were lost prior to cryopreservation and that there was no significant difference among the groups. Post -thaw viability rate was 80-84% and it was the same for all the groups. Motility rate was 70-74%, again with no significant difference among the groups.Conclusion: This sperm cryopreservation method is suitable for oligozoospermia patients because of high recovery and motility rates. It can also help azoospermic patients by preventing repeated surgeries.

Introduction: Morphology and ultra-structure of myoid and sertoli cells were studied in patients with non-obstructive azoospermia who were admitted to testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI), by transmission electron microscopy (TEM). Materials and Methods: 12 biopsies of testicular tissues from patients with non-obstructive azoospermia referred to Royan institute were used as a patient group, another 5 biopsies from person with prostate cancer were used as a control group. The tissues were fixed and processed for TEM, and Sertoli and myoid cells were examined under TEM.Results: Myoid cells of the patient group had lost their slender shape and orientation in relation to the wall of seminiferous tubules. These cells were separated by increased amount of collagen fibers, furthermore, their polarity was different from normal and the nuclear shape changed. Sertoli cells were round to ovoid in shape, and the nuclei were more regular in shape than that of the normal with peripherally located nucleoli.Conclusion: Sertoli cells in the patient group were immature or undifferentiated. This may have resulted in other changes in the testicular tissues such as the alteration in myoid cells and basement membrane, moreover, the immaturity of Sertoli cells may be the cause for the failure in initiating or triggering normal process of spermatogenesis.

Introduction: Garlic is considered to be one of the best diseaseتpreventive foods because of its potent and widespread effects. On the other hand, some cardiovascular disorders including hypertension are accompanied with altered responsiveness of vascular alpha-adrenergic receptors. The purpose of this study was to investigate the effect of aqueousgarlic extract on alpha1-adrenoceptor agonist-induced contraction of rat aorta, aorta ring from rats pretreated with aqueousgarlic extract were used as a model in this study. Materials and Methods: Four and eight weeks after treatment with garlic extract, aortic rings were studied for contractile and relaxation response to phenylephrine (PE) and acetylcholine, respectively.Results: The results showed that the contractile response of aortic rings to PE in garlic treated rats for 4 and 8 weeks decreased (P<0.05), and their endothelium-dependent relaxation response increased in comparison with the controls (P<0.05). The relaxing activity of garlic on rat aorta was time-dependent and this effect was attenuated following endothelium removal. Conclusion: These data suggest that alpha1-adrenoceptor antagonistic action of garlic on rat aorta involves at least two different mechanisms: one is direct relaxing action of garlic on smooth muscle, and the other is indirect and dependent on garlic effect onalpha1-adrenoceptors signal transduction cascade.

Introduction: Eugenol (Eg), a compound extracted from Clove tree, is known for its anti- anaphylactic, anti-inflammation, antiseptic, antibacterial and analgesic properties as well as chain breaking antioxidant in inhibition of lipid peroxidation. Retinoic acid (RA), one of the synthetic derivatives of vitamin A, has been frequently used for treatment of face acne. In our studies Eg was used to test its probable correcting action on decreasing the toxicity and the teratogenic effects of retinoic acid in female NMRI mice embryo. Materials and Methods: Five groups of 8 weeks old pregnant female mice were fed as follows: 1- an oral dose of 60 mg RA per kg b.w. on day 10 of pregnancy, 2- an oral dose of 100 mg/kg b.w. Eg for ten days (5th to 15th day of pregnancy), 3- olive oil (solvent) for 15 days, 4-Eg plus RA (as in groups 1 and 2) and 5- untreated controls. On day 18 of pregnancy the mice were dissectioned and the embryos were taken from the uterus. The length of the Crown Rump (CR), Cranio-Facial (CF), hands, feet and tail were measured and the organs were observed for probable defects. ANOVA , Tukeys and Chi-square were used for data analysis.Results: A positive relationship was observed between the quantity of RA taken by the animals and the frequency of embryonic organ defects. Simultaneous administration of Eg with RA significantly reduced the frequency and the intensity of embryonic defects. RA could significantly reduce the length of hands, feet, tail, CR and CF (p<0.001). It could also cause embryonic defects such as short lower jaw, spiral tail, foot bending, hand and feet oligodactily and syndactily. Simultaneous administration of Eg with RA could significantly correct the length reduction in hands, tail and CF by 16, 18.85, 7.44%, respectively (P<0.001), and CR by 6.57% (P<0.05).Conclusion: The results of this study show that Eg may decrease the teratogenic effects of RA, and might also be suggested as aprotective agent to be used with RA in skin therapy during pregnancy.

Introduction: Heterochromatin consists of DNA sequences that are not transcribed, and are repeated in short tandem at heterochromatic regions of chromosomes 1, 9, 16 as well as the distal part of long arm of chromosome Y. Slightly large tandem repeats at the centromeres of human chromosomes are also considered as heterochromatin regions. The main aim of the present study is to evaluate the heterochromatin polymorphism associated with chronic myeloid and acute non-lymphocytic leukemias.Material and Methods: 16 Leukemic patients and 10 normal healthy persons were selected. By applying Barium Hydroxide Saline Giemsa (BSG) method. The variant heterochromatin of chromosome 1, 9 and 16 on bone marrow samples and lymphocyte cultures were evaluated.Results: the result of present study indicated no significant differences for heteromorphisms between Acute Non-Lymphocytic Leukemia (ANLL) Patients and normal individuals. But differences were significant on comparing the complete inversions in ANLL patients with the controls. There were significant differences in heterochromatin variant of C-segment in chromosomes 1 and 9 between Chronic Myeloid Leukemia patients (CML) and normal group. Partial and complete inversion between CML patients and the control group is significant. Conclusion: A number of reports have indicated pronounced heteromorphism in size and localization in constitutive band region of chromosomes 1, 9 and 16 in many malignancies. However, this study showed similarities and dissimilarities with other investigators regarding heterochromatin variations and inversions.

Introduction: Listeria monocytogenes is a Gram positive bacterium, often found in the environment and is responsible for serious food-borne diseases such as perinatal infections, septicaemia and meningoencephalitis in humans and animals. For this reason, distribution of the ctp A (copper transport protein A) among L. monocytogenes was isolated from clinical, environment, dairy, and poultry samples, and was investigated. Then, ctp A gene was transferred into E.coli DH5-α.Materials and Methods: This investigation was carried out in two steps (ctpA was found in L. monocytogenes isolated from different sources, which was kept in culture collection of Adelaide University, Australia. Then ctpA gene was transferred into E. coliDH5-α. CtpA DNA from L. monocytogenes was amplified by PCR, identified on agarose gel, purified by phenol, and ligated intopGEM-T vector. Then, it was transferred on X-gal plate containing ampicillin. The sequencing of ctpA DNA was determined by using dyeterminator kit to purify DNA, and followed by sequencing machine. Results: By using PCR to identify the homologous DNA in 69 isolates, 38% of tested isolates contained ctpA determinant. Our results showed that: 90% of clinical and dairy isolates; 85% of environmental isolates and 7% of poultry isolates of L. monocytogenes contained ctpA in chromosome DNA. Fortunately, the transformation of ctpA from L. monocytogenes into E. coli DH5-a was successful.Conclusion: Since, the existence of ctpA in clinical, dairy and environmental samples was 90% and in poultry was 7%, there fore, the virulence of all strains of this bacteria are not the same. Introducing such clone (ctpA gene) into suitable carrier strains, could be expected to produce a good oral immunogen against L. monocytogenes.

Introduction: The aim of this study was to determine mouse endometrium glycocalyx cell surface alteration after ovarian hyperstimulation using hMG, hCG and daily progesterone injection at implantation time. Materials and Methods: For this purpose adult NMRI mice were super ovulated using hMG, hCG and then daily injections of progesterone (1 mg/mouse) performed in one group. The animals were sacrificed by cervical dislocation 3.5 and 4.5 days after hCG injection. Tissues were obtained from 1/3 middle part of uterine horns and processed for light and transmission electron microscopic studies.Results: Our results showed that the intensity of PAS reaction on surface epithelium of the control and the hyperstimulated groups increased 4.5 days after mating or hCG injection, but in the hyperstimulated and progesterone injected group there were intense PAS reaction on the 3.5 day after hCG injection. A well defined glycocalyx was observed on the epithelial cell surface of the control group and the hyperstimulated group, there were a lot of long cylindrical microvilli on their epithelium but after progesterone injection some of the microvilli and glycocalyx disappeared and there were some cytoplasmic projections.Conclusion: Thus ovarian hyperstimulation caused alteration in the glycocalyx and these changes may affect the electrical charge of the endometrium at the implantation period. Thus these changes could have influenced endometrial receptivity.

Introduction: L. pneumophila is the most important cause of legionaires, disease, which is currently reported either as a nosocomial or community acquired infection. Rapid and reliable diagnosis of legionella has hampered the epidemiological studies and disease control activities. In spite of good sensitivity and specificity, the culture method encountered serious shortcomings. Polymerase chain reaction (PCR) has been used to detect the viable but non-culturable legionella. In the present investigation, the efficacy and accuracy of mip gene based primers were tested in PCR for culture-negative samples. Materials and Methods: The samples used in this investigation were previously reported as negative by means of conventional culture methods. DNA extraction was carried out by freezing-boiling method. The small fragment of mip gene (of L. pneumophila) was used for the primer set (LEG 1 and LEG-2) in PCR, and also for specific probe LEG-3 in southern blot.Results: From 32 culture-negative samples subjected to these primer sets, L. pneumophila DNA was detected in 6 samples. System accuracy was then checked by dot blot hybridization.Conclusion: The results of this investigation indicated that the PCR method, with suitable primer set and probe, detects the viable but non-culturable Legionella in clinical and environmental samples.