Cytochemical evaluation of sperm chromatin and DNA integrity in couples with unexplained recurrent spontaneous abortions.

The aim of this study was to examine the possible relationship between sperm DNA integrity and chromatin packaging evaluated by cytochemical assays, traditional sperm parameters and recurrent spontaneous abortion (RSA) of unknown origin. In this cohort study, 40 couples with a history of RSA and 40 couples with proven fertility were considered as case and control groups respectively. The semen samples of all husbands were analysed for sperm parameters and also sperm chromatin and DNA integrity assessed using cytochemical tests including aniline blue (AB), chromomycin A3 (CMA3), toluidine blue (TB), acridine orange (AOT) and nuclear chromatin stability assay. Among different sperm parameters, only slow motility was significantly different between the two groups. In sperm chromatin evaluations, there were significant differences between the two groups in all of the tests. In addition, the majority of semen samples in RSA patients exhibited upper percentages of abnormal spermatozoa than the cut-off values regarding different cytochemical assays. Our study showed that in the cases of RSA, slow motility had a significant reduction in comparison with controls and also spermatozoa of men from RSA group had less chromatin condensation and poorer DNA integrity than spermatozoa that obtained from fertile men with no history of RSA.

Beneficial effect of tamoxifen on sperm recovery in infertile men with nonobstructive azoospermia.

About 10% of infertile men have azoospermia. After the introduction of microinjection [intracytoplasmic sperm injection (ICSI)], many of these men obtain the chance to be a father. But still in many cases of nonobstructive azoospermia, we are not able to find spermatozoa for ICSI. Medications may be able to increase the chance of finding spermatozoa in testis samples. So in this study, we evaluated the effect of tamoxifen citrate on the results of sperm recovery from testis tissue in infertile men with nonobstructive azoospermia. Thirty-two azoospermic infertile men with proved nonobstructive azoospermia were selected. Tamoxifen was administered for 3 months. Semen samples and in the cases of azoospermia second testis biopsy were taken, and the results were compared with the first samples. According to first testis samples, 13 patients had hypospermatogenesis, 9 had maturation arrest and 10 patients sertoli cell syndrome. After tamoxifen treatment, six patients showed spermatozoa in their ejaculates. From other patients all in hypospermatogenesis group, 75% in maturation arrest group and 20% in sertoli cell group showed spermatozoa in their second testis samples. Our study showed that treatment of patients with nonobstructive azoospermia with anti-oestrogenic drugs like tamoxifen can improve the results of sperm recovery in testis samples and also increase the chance of pregnancy by microinjection.

Analysis of antimicrobial, antifungal and antioxidant activities of Juniperus excelsa M. B subsp. Polycarpos (K. Koch) Takhtajan essential oil.

Juniperus excelsa M.B subsp. Polycarpos (K.Koch), collected from south of Iran, was subjected to hydrodistillation using clevenger apparatus to obtain essential oil. The essential was analyzed by gas chromatography/mass spectrometry (GC/MS) and studied for antimicrobial, antifungal and antioxidant activities. The results indicated α-pinene (67.71%) as the major compound and α-cedral (11.5%), δ3-carene (5.19%) and limonene (4.41%) in moderate amounts. Antimicrobial tests were carried out using disk diffusion method, followed by the measurement of minimum inhibitory concentration (MIC). All the Gram positive and Gram negative bacteria were susceptible to essential oil. The oil showed radical scavenging and antioxidant effects.

Effect of varicocele on chromatin condensation and DNA integrity of ejaculated spermatozoa using cytochemical tests.

Varicocele occurs in approximately 15% to 20% of the general male population and it is the most common cause of poor semen production and decreased semen quality. It has been demonstrated that patients with varicocele have a significantly higher DNA fragmentation index (DFI) and spermatozoa with nuclear anomalies than healthy fertile men. Therefore, the aim of this study was to evaluate sperm chromatin integrity in these patients. Sixty men referring to the andrology laboratory were categorised into three different groups: 20 infertile men with varicocele, 20 infertile men with abnormal semen parameters and 20 fertile men who had normal spermatogram were considered as control group. Semen analysis was performed according to WHO criteria. To evaluate sperm chromatin quality and DNA integrity, after fixation of sperm smears, aniline blue, toluidine blue, chromomycin A(3) and acridine orange staining were applied in three groups. The slides were analysed by light and fluorescent microscopy and to determine the percentage of mature or immature spermatozoa, 200 spermatozoa were counted in each slide. The results showed that the rates of aniline blue-reacted spermatozoa were significantly higher in infertile and varicocele patients than in the normal group (P < 0.001). In addition, with regard to chromomycin A(3), acridine orange and toluidine blue staining, there was a significant difference between the three groups (P < 0.001). The results showed that the varicocele samples contain a higher proportion of spermatozoa with abnormal DNA and immature chromatin than those from fertile men as well as infertile men without varicocele. Therefore, varicocele results in the production of spermatozoa with less condensed chromatin and this is one of the possible causes of infertility due to varicocele.