Introducing a quantitative method to calculate the rate of primary infertility

In the previous studies, the rate of primary infertility was reported differently. It seems the main reasons are related to the different methods of data collection and information analysis. Therefore, introducing a precise method to determine the infertile couples and the population exposed to the risk of infertility is an important issue to study primary infertility. The proposed methodology for assessing primary infertility rate has been designed and applied by Avicenna Research Institute in a national survey. Sampling was conducted based on probability proportional to size cluster method. In this survey, after reviewing the former studies, the reproductive history was used as a basis for data collection. Every reproductive event was recorded with a code and a date in the questionnaire. To introduce a precise method, all possible events were considered thoroughly and for each situation, it was determined whether these cases should be considered in numerator, denominator or it should be eliminated from the study. Also in some situations where the correct diagnosis of infertility was not possible, a sensitivity analysis was recommended to see the variability of results under different scenarios. The proposed methodology can precisely define the infertile women and the population exposed to the risk of infertility. So, this method is more accurate than other available data collection strategies. To avoid bias and make a consistent methodology, using this method is recommended in future prevalence studies

Testis specific gene 10 expression in the testes of patients with non-obstructive azoospermia

Disorders in the expression of any gene effective in spermatogenic pathway is known as a probable cause of non-obstructive azoospermia and male infertility. The way responsible genes for sperm motility are expressed can considerably affect male fertility. Recent studies show that TSGA10 gene is effective in the natural process of spermatogenesis as protein produced by this gene in mouse results in the production of the main structure of sperm tail. Up to now, no comprehensive studies have been done on the way this gene is expressed in the infertile's testical tissue. In this study, TSGA10 mRNA expression in testicular samples of 84 patients with non-obstructive azoospermia was investigated by semi-quantitative nested RT-PCR in Avesina Infertility Clinic during 2005-6. Moreover, expression levels of TSGA10 during spermatogenesis were evaluated using Johnsen's method for histopathological scoring of the samples. For statistical analysis, SPSS software [Version 11.2] was used. The difference between gene expressions was done based on quantitative variables by the use of t-test and covariance analysis and alpha<0.05 was regarded as a statistically significant value. Testicular TSGA10 mRNA expression was observed in 31 patients, [36.9%], with non-obstructive azoospermia which it had a statistically significant correlation with spermatogenesis progress [p<000.0]. Histopathologically, the gene had been expressed in patients with higher Johnsen's score of spermatogenesis while a lack of expression was seen in all of those with Johnsen's score less than 4.5. The findings indicate that TSGA10 is expressed in human testis and it is restricted to germ cells. It seems that lack of TSGA10 expression may have negative effects on spermatogenesis and on male fertility. On the other hand, determination of the timing of gene expression in a certain level of spermatogenesis may also be used to determine levels of spermatogenesis in azoospermic patients alongside histopathological findings

Polymorphism in phospholipid hydroperoxide glutathione peroxidase [PHGPX] gene in Iranian infertile men

Leukocytes and defective or dead spermatozoa in human semen are a source for the production of reactive oxygen species [ROS] and subsequent injury to intact sperms. Enzymatic and non-enzymatic defensive mechanisms in semen detoxify these compounds. Glutathione peroxidase-4 [GPX-4 or PHGPX] is a major selenoprotein in sperm and it is one of the enzymatic mechanisms that play multiple roles during spermatogenesis. Some of these roles are formation of the mitochondrial capsule, hydroperoxide detoxification and sperm chromatin condensation. Any decrease in the enzyme activity or content, may create disorders in spermatogenesis and sperm fertilizing ability. Considering defects in the expression of the enzyme gene or presence of mutations which may cause decreases in PHGPX activity or content, this study was carried out to identify a number of important mutations in GPX-4 gene by PCR-RFLP method in Iranian infertile men. This study was performed on 128 Iranian men who had been referred to Avesina Infertility Clinic, including 74 infertile men with defective sperm parameters, 18 normozoospermic and 36 fertile subjects as controls. Mean +/- SD for sperm parameters were determined. Genomic DNA was extracted using salting out procedure from peripheral blood leukocytes. PCR-RFLP was done by two sets of primers with 237 bp and 148 bp PCR products that were designed for 1A and 4 exons of GPX-4 gene covering nucleotides of+6 [C[right wards arrow]T], +17 [G[right wards arrow]A], +1725 [G[right wards arrow]A] by Mwol, PshAI and SatI enzymes. Digestion of a 237 bp intact PCR product by Mwol generates two fragments [151 bp and 86 bp]. When a mutation occurs in the restriction site +6 [C[right wards arrow]T], the enzyme would not recognize the sequence, therefore 237 bp segment remains undigested. Treatment of 237 bp segment with PshAI generates two fragments [161 bp and 76] in the intact gene but the same enzyme can not digest 237 bp segment when a mutation occurs in the restriction site +17 [G [right wards arrow] A]. Ultimately, digestion of 148 bp intact segment with SatI generates two fragments [108 bp and 40 bp] but when a mutation occurs in the restriction site +1725 [G [right wards arrow] A], the enzyme will not recognize the sequence; therefore 148 bp segment remains undigested. Enzymatic digestion evaluations of 237 bp and 148 bp segments in all participants revealed that neither of the examined mutations existed in GPX-4 gene. According to the results of this study, it is determined that the prevalence of these mutations in Iranian infertile men is probably low and it may have no association with the etiology of the disorder affecting sperm parameters. Hence, a study with a larger number of patients is suggested to determine the exact prevalence of these and other mutations of the gene in Iranian infertile men

effects of human chorionic gonadotropin on germ cell maturation and testosterone secretion in neonatal mouse testis

Human chorionic gonadotropin [hCG] as an LH agonist affects spermatogenesis and germinal cell numbers, and has extensive usages in infertility treatments. The aim of this study was to determine the effects of varied doses of hCG on germinal cell proliferation and androgenic status in mouse model. In this study, hCG dosages of 5 to 50 IU were injected into 18 mice in three experimental groups and 6 mice served as the control group [Group 1]. The mice in groups 2, 3 and 4 received subcutaneous injections of 5, 10 and 50 IU doses of hCG respectively, on days 15 and 25 of their lives. Blood samples were obtained from each mouse on days 28 and 65 for serum measurements of testosterone. One testis of each mouse was harvested for flow cytometric DNA analysis on day 65. Serum testosterone levels on day 28 were greater in groups 2, 3 and 4 compared to that of the control group. With increasing doses of hCG, the mean testosterone levels increased too and the highest values were observed in group 4. However, serum testosterone levels on day 65 were greatest in group 1 but progressively decreased in groups 2, 3 and 4, lowest in group 4, but there were no significant statistical differences among the groups. Groups 3 and 4 had a significantly reduced mean haploid cell numbers on day 65. The results of this study showed that testosterone production in neonatal mouse testis increases after hCG injection and there is a linear relationship between serum testosterone and hCG injections. With the passage of time and clearance of hCG, Leydig cell stimulation decreases and subsequently testosterone levels diminish too, especially in mice with highest doses of hCG injections. Therefore, for testosterone production in neonatal mouse testis, continuous stimulation of Leydig cells is essential

[Designing an ELISA method using sperm antigens for detection of antisperm antibody in comparison with SpermMar test

The role of antisperm antibodies with a prevalence of 6-26% is well known in immunological infertility. Thus, there is clinical importance to determine ASA levels in both male and female. Nowadays, one of the most important discussed controversies in the field of immunological infertility is establishing an standard method to determine ASA. It seems that ELISA method will be more sensitive, specific and more diagnostic in determination of ASA if sperm surface antigens could be used as coated antigens, with least contamination to sperm intracellular antigens and nonspermic antigens. So, the aim of this study is designing an ELISA method by using the best method of sperm antigens extraction with at least contamination. In this study we designed an ELISA method with three different extraction methods of sperm antigens including sonication method, using SDS detergent, and application of LIS detergent, then we compared ELISA method based on the three extraction methods as well as two similar commercial ELISA kit [IBL Co, and Bioserv Co] with SpermMar test. Comparing designed method with commercial kit indicated that among 28 sera which had 16 positive sera and 12 negative sera by SpermMar, 14 sera were true positive by LIS method and only 2 cases were false negative without any false positive results, whereas there were 5 true positive results and 11 cases false negative by the sonication method. The SDS method also had 13 true positive results with 3 false negative and 4 false positive results. In addition, two commercial kit had in turn 7 and 4 cases true positive and both of them had 1 case false positive and in turn 9 and 12 cases with false negative result. ELISA method designed by LIS detergent has adequate sensitivity [87.5%] with higher specificity [100%] and efficacy [92.8%] than other extraction methods. There is a significant correlation between this designed method and SpermMar test [r=0.572]. The results of this study indicated that ELISA method by LIS antigens has at least contamination with nonspermic antigens and it is better than other extraction methods and commercial ELISA kits for detection of antisperm antibody

[Effects of different media on fallopian tube epithelial cells culture and human]

The advantage of fallopian tube epithelial cell [FTEC] co-culture system on development of human embryo has been shown. However, lack of appropriate media for both FTEC and embryo is one of the disadvantages of this system. The aim of the present study was to determine the effects of different culture media on both FTEC and human embryo development. Healthy fallopian tube samples were removed from 19 previously fertile women under 40 ages undergoing total abdominal hysterectomy. Epithelial cell-rich suspension was prepared mechanically and enzymatically and cultured in three different media [DMEM/F12, RPMI-1640 and Ham's F10]. After four passages, the cells were cultured on plastic to obtain unpolarised confluent cells, or seeded into matrigel to produce polarized cells for 7 days. Viability of cells during and after passages was evaluated by neutral red and trypan blue vital staining. In addition, 117, 45 and 48 surplus 4-8 cell human embryos were cultured in Ham's F10, RPMI and DMEM/F12 for 120 hours, respectively. Their morphology were assessed daily. Cellular vitality during culture in DMEM/F12 was 100%, which was significantly different from that in RPMI and Ham's F10 [95% and 93%, respectively] [P<0.05]. However, after cellular proliferation and passages, no significant difference was observed for different media. Moreover, a significantly higher ratio of embryos reached the morula stage in Ham's F10 [79.4%] than RPMI [68.8%] and DMEM/F112 [62.5%] [P<0.05]. It is concluded that DMEM/F12 was superior to other media during cell proliferation and passage of FTEC. However, after cell passage and during co-culture of embryos with FTEC, Ham's F10 was a more appropriate medium than the other two

Psychosocial aspects of infertility from viewpoint of infertility treating physicians

Infertility phenomenon extra to medical science territory has been studied in the fields of behavior and social sciences. Infertility as a psychological crisis, has a lot if stress on infertile couples and in different ways, has threatened their mental health. Considering high expenses of fertility in Iran with its socio-cultural aspects, especially for women, has propounded special importance for studying of infertility phenomenon from psychological-social aspects. This research, as preliminary study, has studied viewpoint of Iranian physicians and specialists involved on infertility which have beneficial experiences in regard if infertile couples, as one if most important and reliable resources for cultural-social studies of infertility. This study is a purposive research which by design of psychological-social primary problems in infertility, it investigate the recognition if role if psychological-social factors from view point of Iranian physicians and recognition rate of importance of psychological-social problems with these physicians. In this way infertile couples get more psychological-social supports and also in respect of economical it prevent expending unnecessary time and expenses or performing improper medical treatments and it can accompany the medical treatments with more effective results. After validity inspection of sandbi questionnaire [1997] in primary study, for studying psychological-social factors involved in infertility, questionnaire with 40 questions was prepared and was delivered to 120 physicians and specialist of different fields involved in infertility affairs [Obs and Gyn, Urology, Embryology, Infectious disease and General practitioner]. Findings of this descriptive research showed that infertile on the viewpoint of Iranian physicians of infertile community in Iran, which needs more attention. Also, from their point of view, infertile communities belong to families with low income and infertility treatments enforce high expenses to them. Most common emotional and psychological problems of infertile couples were despair, frustration, fear and anxiety. It is has been accompanied in less cases with rage and violence. These problems, with familial and social problems, such as suspension of marital relations and conflicts such as second marriage, separation and divorce are of important matters, which make the psychological consultation services very important and vital. Hopelessness and despair, fear and anxiety, are important psychological factor diseases. Women are more than men patent to psychological hams in this regard. In view point of physicians, stress and glumness in causing infertility, consultation and psychological treatment and awareness of details of diagnostic and remedial proceedings in treatment and improvement of infertility, has effective role and presence of psychiatrist and consultant seem to be necessary in different treatment and clinical infertility wards. In this way, necessary psychological interventions will be provided in all process of before, during and after treatments