Evaluation of infertility indicated for women failed to conceive after one year of unprotected intercourse and for women over 35 years failed to conceive after 6 mo of unprotected intercourse, because the fertility decline as women approach 40 years. Evaluation of infertile couple should begin with semen analysis of male partner. If the semen analysis is within normal range, evaluation then move on to female partner. Evaluation of the infertile woman should be carried in cost-effective manner to identify the causes of infertility using the least invasive methods. Methods of evaluation of infertile women include: 1) history and physical examination; 2) evaluation of the male partner; 3) documentation of ovulation; 4) evaluation of the ovarian reserve; 5) evaluation of cervical causes of infertility; 6) evaluation of uterine causes of infertility; 7) documentation of the tubal patency; and 8) exclusion of peritoneal causes of infertility. Evaluation of infertile couple should include evaluation of both partners. Semen analysis for the male partner, followed by documentation of ovulation and evaluation of genital tract patency. PCT is not used as routine evaluation of infertile women. Laparoscopy is not used as routine evaluation of infertile women unless there is suspected peritoneal factors of infertility or endometriosis or tubal occlusion. Ovarian reserve should only be done for infertile women with diminished response to external gonadotropins (not routine).

Objective: To evaluate the effect of Quercetin (QT) on erectile dysfunction and oxidative stress in penile tissue of streptozotocin-induced diabetic rats. Methods: Two weeks after diabetes induction, QT was treated to normal and diabetic rats for 5 wk. Sexual behavioral parameters including mount latency, intromission latency, ejaculation latency, post-ejaculatory interval, mount frequency and intromission frequency, were observed against stimulus females. Sperm count and their motility and viability were recorded. Serum glucose and testosterone levels were estimated. In penile tissue levels of cyclic guanosine monophosphate, thiobarbituric acid reactive substances and glutathione, and enzymatic activities of superoxide dismutase and catalase were measured. Histopathological changes were evaluated in a cross-section of penile tissue. Results: Sexual behavioral ejaculation latency, post-ejaculatory interval, mount latency and intromission latency were significantly increased while mount frequency and intromission frequency were decreased in diabetic rats. Treatment with QT corrected the male sexual behavioral levels and also enhanced the inhibited sperm count, motility and viability in diabetic rats. Serum testosterone and penile cyclic guanosine monophosphate levels were significantly increased in QT treated diabetic rats compared to untreated diabetic animals. Penile oxidative stress biomarkers were corrected by the QT treatments in diabetic rats. Histopathological evaluation revealed damaged penile tissues in diabetic rats, which was protected following QT treatment. Conclusions: QT eliminated the diabetic-induced sexual impairment and showed significant antioxidant effects in penile tissue. Further experimental studies are recommended for QT therapeutically usage.

Objective: To evaluate the effects of pellucid zone 3 monoclonal antibodies against B-cell lymphoma 2 (BCL-2) expression and mice follicle diameter at various time periods. Methods: The animal model of this study was 36 Balb/c mice (Mus musculus). A true experimental design was used with a post-test only control group approach. BCL-2 expression was observed using immunohistochemistry, while the follicle diameter was observed by haematoxylin-eosin staining. The data was analyzed using nested ANOVA to compare the results of the mean expression of BCL-2 on the 5th and 20th day of observation in the pre-antral and antral follicle between the control and treatment groups. Results: No significant differences were found in BCL-2 gene expression. There were also no significant differences in BCL-2 expression on the 10th day of pre-antral follicle analysis. Moreover, there were no significant differences between the mean follicle diameter on the 5th, 10th, and 20th day of pre-antral and antral follicle development between the control and treatment groups. The addition of bovine pellucid zone 3 (bZP3) monoclonal antibodies on the 5th and 20th day of observation did not decrease the expression of BCL-2 gene in the pre-antral and antral follicle of mice. Administering bZP3 monoclonal antibodies on the 10th day of observation did not affect BCL-2 expression in the pre-antral follicle but did decrease BCL-2 expression in the antral follicle. Supplying bZP3 monoclonal antibodies on the 5th, 10th and 20th day did not affect the diameter of pre-antral and antral follicles of the mice. Conclusion: The monoclonal antibodies bovine zona pelusida 3 has the potential to be developed as a safe immunocontraception preparation.

Objective: To investigate if the protective effect of α-tocopherol against the impact of ethanol on brain morphogenesis involved the activity of the selenoproteins phospholipid hydroperoxide glutathione peroxidase (PHGPx; GPx4) and selenoprotein P (SelPP) that have roles against oxidative stress. Methods: Forty female mice were randomly assigned into natural control (CON), positive control (ETOH), low-, medium-, and high-α-tocopherol-supplemented-ethanol groups (LTOC, MTOC, HTOC, respectively). CON received drinking water without ethanol while ETOH, LTOC, MTOC and HTOC groups received 20% ethanol in drinking water. The supplemented groups were given respective dosages of α-tocopherol, 0.410, 0.819, and 1.640 mg/g body weight, at day 14 before mating onwards to the day 9 of gestation. At 10.5 ED of gestation (1 100 h), the pregnant females were sacrificed by cervical dislocation and the embryos were harvested. Total RNA were extracted, cDNA synthesis and qRT-PCR analyses were carried out. Results: The level of expression of PHGPx in the positive control was significantly lower than that of the natural control. Among the three α-tocopherol-supplemented groups, only the medium dose-group was significantly higher than the positive control. The level of expression of SelPP in the positive control was significantly lower than those of the natural control, the low- and medium-dose-tocopherol supplemented groups. In the high dose-α-tocopherol supplemented group, the level of expression was not significantly different from the positive control but significantly lower than the natural control. Conclusions: The activity of the selenoproteins PHGPx and SelPP are involved in the internetwork of antioxidative enzymes with vitamin E when given up to a medium dose only and is one of the possible pathways of shielding embryonic development against the impact of ethanol on brain morphogenesis. This study strengthens the impact of dietary α-tocopherol and Selenium supplement during the critical period of pregnancy.

Objective: To establish ultrasound guided testicular fine needle aspiration (TFNA) as well as to assess the effectiveness of uni-directional (UD) and multi-directional (MD) TFNA in buck according to testicular cells, echotexture and gross changes of testicle, age of buck was considered. Methods: A total of 120 samples were collected with both directions (UD, n=60) and (MD, n=60) suction from testis of 10 apparently healthy bucks. All slides were stained with May-Grünwald-Giemsa and examined under light microscope with 1 000 × magnifications to count spermatogenic cells, spermatozoa and sertoli cells. The percentage of spermatozoa and sertoli cells were expressed as spermatic index and sertoli cell index. Results: Results revealed no difference in the presence of various spermatogenic and sertoli cells in cell cluster of slides made either unidirectional TFNA or multidirectional TFNA. Early spermatids were the most numerous, followed by late spermatids, primary spermatocytes, spermatogonia. Sertoli cell index was higher in TFNA smears of young bucks prepared 7-13 mo of age and spermatic index was higher in adult bucks 14-24 mo of age. No echogenic change was observed in the echotexture of testisafter TFNA. Conclusions: It seems that TFNA has no serious ill effect on the buck testis when uni-direction aspiration is performed. Moreover, the possibility to standardize this method might provide a greater impulse to the clinical diagnostics of male animal infertility.

Objective: To evaluate the effects of nano-particle of selenium (nSe) on plasma antioxidant capacity, sexual and thyroid hormones and spermatogenesis in male rats exposed to oxidative stress. Methods: Forty rats were randomly divided into four treatments with ten replicates. Treatment groups were: C, the control group received normal saline as gavage and injection (i.p.); OS, received tert-butyl hydroperoxide (0.2 mmol/kg body weight) for inducing oxidative stress; nSe, received nSe (0.3 mg/kg body weight) as gavage, and OS+nSe, received tert-butyl hydroperoxide and nSe. All groups were treated for 28 d and administrations were done each 48 h. Results: Oxidative stress decreased and gavage of nSe to stressed rats increased the antioxidant capacity and activities (P<0.05). Oxidative stress decreased and nSe increased the levels of thyroid, gonadotropic and testosterone hormones as compared with the control group (P<0.05). Gavage of nSe to stressed rat resulted in increase (P<0.05) of gonadotropin and testosterone hormones compared to oxidative stress group. There were no significant differences (P>0.05) between rats exposed to oxidative stress and those in the control group for sperm quantity and quality. Gavage of nSe to stressed rats had no effect (P>0.05) on the sperm parameters, except increased viability and progressive percentages. Conclusions: Nano-particle of Selenium administration in stressed rats could ameliorate the negative effects of oxidative stress on the antioxidant capacity and activities, but not on the quantity and quality parameters of sperm.

Objective: To evaluate the effect of adding different values of polymyxin B (PMB) to bull semen on various motility parameters of post-thawed semen such as total motility, progressive motility and velocity parameters using kinetic parameters of sperm by Computer Assisted Sperm Analysis. Methods: Gram negative bacteria release lipopolysaccharide, which induces the apoptotic pathway. Antibiotics are added to semen in order to prevent bacterial contaminations in bovine semen. These antibiotics kill the bacteria especially gram negative bacteria. Therefore, their endotoxins are released during bacteriolysis and bind to the head region and midpiece of sperm. PMB is a bactericidal antibiotic against multidrug resistant gram-negative bacteria and is able to neutralize the toxic effects of the released endotoxin. This study was performed on 3-year old Taleshi bulls. Results: The results showed both positive and negative significant effects of PMB on semen quality. Total motility and progressive motility were significantly increased (P<0.000 1) by 100 μg per mL of PMB (55.2% and 48.8% respectively) against the control groups (43.5% and 37.7%, respectively). Moreover, they were significantly decreased (P<0.000 1) by 1 000 μg per mL of PMB (35.2% and 28.8% respectively) against the control groups (43.5% and 37.7% respectively) in above-mentioned parameters. In Computer Assisted Semen Analyzer, parameter VAP was significantly decreased (P<0.04) in 1 000 μg (69.6 μm/s) against the control group (78.7 μm/s). Finally, using PMB in processing cryopreserved bull semen is advised, but before using it, the rate of endotoxins must be measured. Conclusions: We advise using PMB after measuring endotoxin concentration; In vitro, in vivo and in field fertilization, adding other sperm evaluation factors such as acrosomal integrity, DNA integrity, mitochondrial function to PMB treated semen.

Objective: To assess the effects of epidermal growth factor (EGF) on the Egyptian buffalo bull frozen semen, EGF was incorporated at 0 (control), 50, 100, 200 and 400 ng/mL of extender (Bioxcell®). Methods: Semen features, spermatozoa biometry, total liberated amounts of enzymes (aspartate transaminase, alanine aminotransferase, lactate dehydrogenase, acid phosphatase and alkaline phosphatase) and lipid peroxidation markers (thiobarbituric acid reactive substances, malondialdehyde, glutathione peroxidase, nitric oxide, catalase (CAT) and superoxide dismutase (SOD)) were determined in the spermatozoa-free extracellular extender. Results: Spermatozoa membrane integrity significantly (P<0.05) increased, but DNA integrity decreased with EGF 200 ng/mL. Spermatozoa head (dimensions, area and perimeter), but not shape, as well as acrosome and midpiece measures substantially differed with regard to EGF. Principle piece length and volume markedly decreased (at 100 and 200 ng/mL), while total tail/flagellum length increased (at 50 ng/mL) after EGF supplementation. EGF 50 ng/mL was associated with the decline of nitric oxide levels and catalase enzyme activity, but EGF 100 ng/mL significantly decreased the total liberated amounts of enzymes (aspartate transaminase, lactate dehydrogenase, acid phosphatase and alkaline phosphatase) as well as lipid peroxidation markers (thiobarbituric acid reactive substances and malondialdehyde). Conclusions: EGF in vitro supplementation would affect the semen characteristics of buffalo bull with 100 ng/mL counteracted the freezing mediated oxidative stress indicated with the lowest enzymes leakage and lipid peroxidation.