ABSTRACT
Background: The most common chromosomal abnormality due to non-obstructive azoospermia [NOA] is Klinefelter syndrome [KS] which occurs in 1-1.72 out of 500-1000 male infants. The probability of retrieving sperm as the outcome could be asymmetrically different between patients with and without KS, therefore logistic regression analysis is not a well-qualified test for this type of data. This study has been designed to evaluate skewed regression model analysis for data collected from microsurgical testicular sperm extraction [micro-TESE] among azoospermic patients with and without non-mosaic KS syndrome
Materials and Methods: This cohort study compared the micro-TESE outcome between 134 men with classic KS and 537 men with NOA and normal karyotype who were referred to Royan Institute between 2009 and 2011. In addition to our main outcome, which was sperm retrieval, we also used logistic and skewed regression analyses to compare the following demographic and hormonal factors: age, level of follicle stimulating hormone [FSH], luteinizing hormone [LH], and testosterone between the two groups
Results: A comparison of the micro-TESE between the KS and control groups showed a success rate of 28.4% [38/134] for the KS group and 22.2% [119/537] for the control group. In the KS group, a significantly difference [P<0.001] existed between testosterone levels for the successful sperm retrieval group [3.4 +/- 0.48 mg/mL] compared to the unsuccessful sperm retrieval group [2.33 +/- 0.23 mg/mL]. The index for quasi Akaike information criterion [QAIC] had a goodness of fit of 74 for the skewed model which was lower than logistic regression [QAIC=85]
Conclusion: According to the results, skewed regression is more efficient in estimating sperm retrieval success when the data from patients with KS are analyzed. This finding should be investigated by conducting additional studies with different data structures
ABSTRACT
Background: the sertoli cells in the testis create unique and safe environment to protect seminiferous tubules from auto antigens and invading pathogens. These cells produce the survival factor of the blood-testis barrier and produce special materials such as androgen binding proteins and contribute to the coordinated action of spermatogenesis. Given that the sertoli cells play an essential role in spermatogenesis and the lack of these cells leads to the disruption of spermatogenesis, it is necessary to investigate the behavior and performance of these cells. To achieve this goal, the cells must first be extracted. The aim of this study was to develop a procedure to isolate, culture, and characterize human sertoli cells
Methods: in order to isolate the sertoli cells of azoospermia patients who underwent [testicular sperm extraction] TESE surgery, washing up and multi_stage enzyme digestion of single cells, culture on petri dishes impregnated with datura stramonium lectin agglutinin [DSA] were done and then the cells were passaged for several times and isolated. For more purification, flow cytometry method with FSH receptor antibody was used. Immunocytochemistry assays and Elisa test for identification of these cells were employed
Results: the purification method resulted in more than 97% purity. The nature of sertoli cells was confirmed by morphology evaluation, detecting anti-mullerian hormone in sertoli cell culture media and the presence of FSH receptor on sertoli cells
Conclusion: this study introduced and applied a method to isolate, culture, and purify human sertoli cells with high purity which made possible further researches on these cells
ABSTRACT
Background: The availability of testis specific genes will be of help in choosing the most promising biomarkers for the detection of testicular sperm retrieval in patients with non-obstructive azoospermia [NOA]. Testis specific chromodomain protein Y 1 [CDY1] is a histone acetyltransferase which concentrates in the round spermatid nucleus, where histone hyperacetylation occurs and causes the replacement of histones by the sperm-specific DNA packaging proteins, TNPs and PRMs
Objective: The aim was to evaluate CDY1 gene as a marker for predicting of successful sperm retrieval in NOA patients
Materials and Methods: This research was conducted on 29 patients with NOA who had undergone testicular sperm extraction [TESE] procedure. NOA patients were subdivided into patients with successful sperm retrieval [NOA+, n=12] and patients with unsuccessful sperm retrieval [NOA-, n=17]. Relative expression of CDY1 gene and chromatin incorporation of CDY1 protein were measured by quantitative real-time polymerase chain reaction [qRT-PCR] and ELISA assay, respectively
Results: Quantification of mRNA relative expression and incorporation of CDY1 protein in chromatin showed significant lower expressions and protein levels of CDY1 in testis tissues of NOA- in comparison to NOA+ group
Conclusion: The findings in this study demonstrated a correlation between the low levels of CDY1 function and unsuccessful sperm recovery in the testicular tissues of NOA- compared to NOA+ patients. Therefore, it can be reasonable to consider CDY1 as a potential biomarker for predicting the presence of spermatozoa, although the claim needs more samples to be confirmed
ABSTRACT
Background: although the effectiveness of ginger as an antioxidant agent has been exploited, little human research has been conducted on its activity on male reproductive functions
Objective: this study was designed to investigate the effects of ginger [Zingiber officinale] on sperm DNA fragmentation [SDF] in infertile men
Materials and Methods: this randomized double-blind, placebo-controlled trial with a 1:1 allocation was performed on 100 infertility treatment candidates who were admitted to Royan Institute for Reproductive Biomedicine, Tehran, Iran. Patients were randomly assigned to receive one of two treatments: ginger and placebo. Patients were given a 3-month oral treatment [members received capsules containing 250 mg of ginger powder twice a day in ginger and a placebo in other group]. Before and after treatment, standardized semen samples were obtained to determine sperm concentration, motility, and SDF according to World Health Organization
Results: there was no significant difference between two groups regarding SDF at baseline [53.48. 95%CI: 37.95-69.02] in cases and [56.75, 95%CI: 40.01-73.5] in controls. The average positive percentage of SDF in patients receiving ginger [17.77, 95%CI: 6.16-29.39] was lower compared with placebo [40.54, 95%CI: 23.94-57.13] after three month of treatment [p=0.02]. In multivariate analysis, SDF was significantly lower in patients receiving ginger compared with placebo [mean difference: 3.21, 95%CI: 0.78-5.63, p=0.009]. There were no significant differences between two groups regarding to semen parameters
Conclusion: the present study has demonstrated that ginger in a controlled study of efficacy was effective in decreasing SDF in infertile men
ABSTRACT
To evaluate predictive factors of successful microdissection-testicular sperm extraction [MD-TESE] in patients with presumed Sertoli cell-only syndrome [SCOS]. In this retrospective analysis, 874 men with non-obstructive azoospermia [NOA], among whom 148 individuals with diagnosis of SCOS in prior biopsy, underwent MD-TESE at Department of Andrology, Royan Institute, Tehran, Iran. The predictive values of follicle stimulating hormone [FSH], luteinizing hormone [LH], and testosterone [T] levels, testicular volume, as well as male age for retrieving testicular sperm by MD-TESE were analyzed by multiple logistic regression analysis. Testicular sperm were successfully retrieved in 23.6% men with presumed SCOS. Using receiver operating characteristic [ROC] curve analysis, it was shown that sperm retrieval rate in the group of men with FSH values >15.25% was 28.9%. This was higher than the group of men with FSH
Subject(s)
Humans , Male , Microdissection , Testis , Spermatozoa , Follicle Stimulating Hormone , Luteinizing Hormone , Sperm Retrieval , Retrospective Studies , AzoospermiaABSTRACT
Male infertility is a multifactorial disorder, which affects approximately 10% of couples at childbearing age with substantial clinical and social impact. Genetic factors are associated with the susceptibility to spermatogenic impairment in humans. Recently, SEPT12 is reported as a critical gene for spermatogenesis. This gene encodes a testis specific member of Septin proteins, a family of polymerizing GTP-binding proteins. SEPT12 in association with other Septins is an essential annulus component in mature sperm. So, it is hypothesized that genetic alterations of SEPT12 may be concerned in male infertility. The objective of this research is exploration of new single nucleotide polymorphism G5508A in the SEPT12 gene association with idiopathic male infertility in Iranian men. In this case control study, 67 infertile men and 100 normal controls were analyzed for genetic alterations in the active site coding region of SEPT12, using polymerase chain reaction sequencing technique. Fisher exact test was used for statistical analysis and p<0.05 was considered as statistically significant. Genotype analysis indicated that G5508A polymorphic SEPT12 alleles were distributed in three peaks of frequency in both control and diseases groups. Categorization of the alleles into [GG], [GA], [AA] types revealed a significant difference between infertile patients [azoospermic and asthenospermic] and normal controls [p=0.005]. According to our finding we suggest that G5508A polymorphism in SEPT12 gene can affect spermatogenesis in men, the opinion needs more investigation in different populations
ABSTRACT
Cytomegalovirus [CMV] is a prevalent infection in humans. Recent studies have shown the role of CMV infection in male infertility disorder. Here we aimed to study the role of CMV infection in men with idiopathic infertility. We performed a case-control study of CMV serology in 200 patients attending male infertility clinic of a university hospital. There were 154 men diagnosed with infertility and 46 men without infertility. The patients were asked to donate their sperm, blood, and urine. The presence of CMV infection was studied using quantitative polymerase chain reaction. CMV infection was present in 25 of all the studied participants. Controls had a higher sperm count and sperm motility and sperm morphology compared to patients. There were no significant differences in the studied variables between those with and without CMV infection, nor in patients, neither in controls. Sperm morphology was negatively correlated with cigarette smoking [r=-0.15; p<0.03]. Even though the prevalence of CMV infection was higher in patients with infertility in control and patient [5/46 vs. 20/154] respectively, this was not statistically significant. We did not show a significant role for CMV infection in male infertility. Based on the previous studies, it could be assumed that CMV infection is an important part of the male infertility and its treatment would improve the sperm quality, however this was not confirmed by the present study
ABSTRACT
Septins are an evolutionary conserved group of GTP-binding and filament-forming proteins that have diverse cellular roles. An increasing body of data implicates the septin family in the pathogenesis of diverse states including cancers, neurodegeneration, and male infertility. The objective of the study was to evaluate the expression pattern of Septin14 in testis tissue of men with and without spermatogenic failure. The samples retrieved accessible random between infertile men who underwent diagnostic testicular biopsy in Royan institute. 10 infertile men with obstructive azoospermia and normal spermatogenesis and 20 infertile men with non-obstructive azoospermia were recruited for real-time reverse transcription [RT]-PCR analysis of the testicular tissue. Total RNA was extracted with trizol reagent. Comparison of the mRNA level of septin14 revealed that in tissues with partial [n=10] or complete spermatogenesis [n=10], the expression of septin 14 was significantly higher than sertoli cell only tissues. The testicular tissues of men with hypospermatogenesis, maturation arrest and sertoli cell only had lower levels of septin 14 transcripts than normal men. These data indicates that Septin 14 expression level is critical for human spermatogenesis
Subject(s)
Humans , Male , Spermatogenesis/genetics , Infertility, Male/genetics , Reverse Transcription , Testis/metabolism , RNA, Messenger , Azoospermia/genetics , Gene Expression Regulation , Reverse Transcriptase Polymerase Chain Reaction , Sertoli Cells/metabolismABSTRACT
Intrauterine insemination [IUI] is one of the most common methods in infertility treatment, but its efficiency in infertile couples with male factor is controversial. This study is a retrospective study about correlation between semen parameters and male and female age with successful rate of IUI in patients attending to Royan Institute. A total of 998 consecutive couples in a period of 6 months undergoing IUI were included. They were classified into two groups: couples with successful and unsuccessful pregnancy. Main outcome was clinical pregnancy. Data about male and female ages and semen analysis including concentration, total sperm motility, class A motility, class B motility, class A+B motility and normal morphology was extracted from patients' records. Semen samples were collected by masturbation or coitus after 2 to 7 days of abstinence. Their female partners were reported to have no chronic medical conditions and have normal menstrual cycles. One hundred and fifty seven of total 998 cycles [15.7%] achieved pregnancy. The average of female age in successful and unsuccessful group was 28.95 +/- 4.19 and 30.00 +/- 4.56 years, respectively. Mean of male age was 33.97 +/- 4.85 years in successful group and 34.44 +/- 4.62 years in unsuccessful group. In successful and unsuccessful groups, average of sperm concentration was 53.62 +/- 38.45 and 46.26 +/- 26.59 [million sperm/ml], normal morphology of sperm was 8.98 +/- 4.31 [%] and 8.68 +/- 4.81 [%], sperm total motility was 47.24 +/- 18.92 [%] and 43.70 +/- 20.22 [%] and total motile sperm count was 80.10 +/- 63.61 million and 78.57 +/- 68.22 million, respectively. There was no significant difference in mean of females' age and males' age between successful and unsuccessful groups [P<0.05]. In addition, there was no significant difference in semen parameters including concentration, total sperm motility, class A motility, class B motility, class A+B motility and normal morphology between two groups. It was shown that common semen analysis and male and female ages cannot predict IUI outcome.
Subject(s)
Humans , Male , Female , Infertility/therapy , Pregnancy Outcome , Pregnancy Rate , Retrospective Studies , Semen Analysis , Sperm MotilityABSTRACT
In patients with non-obstructive azoospermia [NOA], vital spermatozoa from the tissue is obtained from testes by enzymatic treatment besides the mechanical treatment. To increase the sperm recovery success of testicular sperm extraction [TESE], with enzymatic digestion if no sperm is obtained from testis tissue by mechanical method. Tissue samples were collected from 150 men who presented with clinical and laboratory data indicating NOA by means of TESE and micro dissection TESE methods. Initially, mature spermatozoa were examined for by mechanical extraction technique shredding the biopsy fractions. In cases whom no spermatozoa was observed after maximum 30 min of initial searching under the inverted microscope, the procedure was followed by enzymatic digestion using DNaseI and collagenase type IV. Surgery type, pathology, AZF, karyotype, hormones and testis size were compared in patients. Of 150 cases with NOA, conventional mincing method extended with enzymatic treatment yielded successful sperm recovery in 13 [about 9%] patients. Comparison of parameters revealed that level of FSH and LH were significantly different [p=0.04 and 0.08 respectively] between two groups that response negative and positive to enzymatic digestion. The combination of conventional TESE and enzymatic digestion is an effective method to recover spermatozoa. The benefit of the mincing combined with enzyme to sperm retrieval for NOA firstly shorten the mechanical searching time, leading to minimizing further cellular damage as well as exposure to external conditions, and secondly reduce the number of cases with sperm recovery failures. Also, the serum level of FSH and LH are factors that influence the chance of sperm retrieval
Subject(s)
Humans , Male , Azoospermia/therapy , Spermatozoa/cytology , Testis/cytology , Sperm Injections, Intracytoplasmic , Microdissection , Biopsy/methodsABSTRACT
To determine the differences in sperm quality and results of intracyto-plasmic sperm injection [ICSI] cycles between three groups of male factor infertile couples: oligozoospermic, obstructive azoospermic and non-obstructive azoospermic. In this prospective cohort study, 628 male factor infertile couples who underwent ICSI cycles from April 2004 to March 2006 were enrolled. Three hundred fourteen oligozoospermic patients [group I], 180 obstructive azoospermic patients [group II] and 134 non-obstructive azoospermic patients [group III] were included. Fertilization, cleavage, implantation and clinical pregnancy, early abortion rates were assessed. Chi-square and analysis of variances with Post Hoc [Tukey test] were used for data analysis. Fertilization rates were significantly different in the three groups [group I: 66.6%; group II: 51.8%; group III: 47.7%; p=0.004]. There were differences in the implantation rates [I: 19.5%; II: 17.6%; III: 6.4%; p=0.001]. The cleavage rates were found to be 55.1% [group I], 47.5% [group II], 45.5% [group III], respectively. The clinical pregnancy rate was the lowest in the third group [I: 37.6%; II: 28.9%; III: 13.4%; p=0.001]. There was no significant difference in early abortion rates between the three groups: [I: 10.7%; II: 9.8%; III: 8%; p=0.776]. It can be concluded that patients with oligozoospermia may benefit the most from ICSI treatment. ICSI cycles which use spermatozoa from non-obstructive azoospermic patients have a lower chance for successful outcome. The results of this study suggest, in cases of failure to achieve pregnancy after 1 or 2 cycles in non-obstructive azoospermic patients, embryo donation would be a better alternative
Subject(s)
Humans , Male , Female , Adult , Azoospermia , Oligospermia , Infertility, Male , Treatment OutcomeABSTRACT
Infertility can be a major concern for couples trying to conceive, and occupational hazards may constitute a main cause of infertility in men. Studies conducted throughout the world indicate that physical and chemical hazards in the workplace can have a negative impact on male fertility. The main objective of this study was to determine the frequency of occupational categories of men who attended an infertility clinic, and to evaluate the differences in the semen quality parameters among occupational categories. This cross-sectional study was conducted on 1164 males who were referred to the Infertility Research Center in Tehran for treatment of infertility in order to evaluate the effects of certain occupations on infertility. The participants were divided into several categories according to their occupations and evaluated by means of a questionnaire for duration of infertility, BMI, sperm count, percentage of normal sperm morphology and percentages of sperm with class A and class B motilities. Descriptive statistics, analysis of variance, and correlations were conducted using SPSS 16.0 for Windows. There were no statistically significant differences in the mean sperm count or sperm morphology between occupational categories. Assessment of the differences in the frequency of sperm motility classes between occupational categories revealed a significant difference only in the frequency of sperm with class B motility. The lowest mean percentages of sperm with class B motility were seen in those involved in the transportation industry, a finding in agreement with a number of other researches. Our findings revealed an association between occupation and sperm motility. Since our study population was relatively small and in many cases exposures to work hazards were brief, a larger study group must be evaluated in order to support the preliminary results of this study
ABSTRACT
The present study offers our contribution on the topic by a retrospective analysis of the prevalence of chromosomal abnormalities in a population of Iranian infertile men attending assisted reproduction programs. Cytogenetic analysis was performed according to standard methods on cultured cells obtained from the patient peripheral blood. In all, 874 files belonging to male partner of each couple were classified as follows: azoospermic, oligozoospermic and patients with low sperm quality in respect of morphology and motility. Chromosomal abnormalities were observed in 136[15.5%] individuals of the whole population studied including 12.0%, 1.2% and 2.0% of azoospermic, oligozoospermic and patients with low sperm quality, respectively. Of those, 116 [13.2%] had sex chromosome abnormalities and 20[2.3%] had autosomal chromosome abnormalities. We observed high frequency of aneuploidy and sex chromosomal mosaicism in azoospermic men and high structural aberrations in males with low sperm quality. We suggested that type of chromosomal abnormalities had an inverse relation to sperm count. So that, high chromosomal aneuploidy was detected in males with lower sperm count and high structural aberration was detected in males with low sperm quality. Chromosomal abnormalities are a major cause of male infertility. Consequently, Genetic testing and counselling is indicated for infertile men with abnormal semen parameters with either abnormal karyotype or normal karyotype before applying assisted reproductive techniques