MicroRNAs association with azoospermia, oligospermia, asthenozoospermia, and teratozoospermia: a systematic review.

Infertility is a major health problem across the world. One of the main reasons for male infertility are defects in sperm. Semen analysis is the most common test utilized to evaluate male fertility and since it suffers from multiple drawbacks, reproduction scientists have tried to find new molecular markers for detecting sperm defects. MicroRNAs (miRNAs) are small molecules in cells which take part in regulating gene expression. Various studies have confirmed miRNAs to have a role in defining multiple sperm characteristics, including sperm count, motility, and morphology. In this paper, we have systematically reviewed the role of miRNAs in infertile men with sperm defects including azoospermia, oligospermia, asthenozoospermia, and teratozoospermia. Also, we have assembled various bioinformatics tools to come up with a pipeline for predicting novel miRNAs which could possibly participate in sperm count, motility, and morphology. Also, related KEGG and GO terms for predicted miRNAs have been included in order to highlight their role in sperm function. Our study emphasizes the potential role of miRNAs in male infertility and provides a general overview for future studies aiming to find robust molecular markers for this condition.

[Distribution of traditional Chinese medicine syndrome types of male infertility in Chengdu area].

OBJECTIVE: To explore the distribution characteristics of the traditional Chinese medicine (TCM) syndrome types of male infertility in Chengdu area, China, in order to provide some objective evidence for the clinical treatment and studies of male infertility. METHODS: We collected the clinical data on 500 cases of male infertility treated in the outpatient and inpatient departments of the Affiliated Hospital of Chengdu University of Chinese Medicine and Chengdu Hospital for Reproduction, Women and Children from January to December 2017. Based on the results of examinations using the four diagnostic methods of TCM, namely, observation, auscultation and olfaction, interrogation, and palpation, we differentiated the TCM syndromes of the patients and analyzed the distribution characteristics of the syndrome types. RESULTS: Analysis of the baseline characteristics and the results of Chi-square test showed statistically significant differences in the distribution of the frequency of sexual intercourse, body mass index, history of urinary tract infection, testis volume, testicular texture, vas deferens and varicocele among the 500 patients (P < 0.05). As for the distribution of the TCM syndrome types, 115 cases (23.0%) were diagnosed with the unlicensed discernible type of syndrome, 109 (21.8%) with kidney-yang deficiency, 36 (7.2%) with kidney-essence deficiency, 30 (6.0%) with both kidney-yang deficiency and liver-qi stagnation, 30 (6.0%) with both kidney-yang deficiency and spleen-asthenia with excessive dampness, 28 (5.6%) with kidney-yin deficiency, 20 (4.0%) with spleen-asthenia and excessive dampness, 19 (3.8%) with liver-qi stagnation, 19 (3.8%) with phlegm dampness obstruction, 19 (3.8%) with kidney-yang and kidney-essence deficiency, 16 (3.2%) with downward damp-heat, 11 (2.2%) with both kidney-yin and kidney-yang deficiency, 10 (2.0%) with qi-stagnation and blood stasis, and 38 (7.6%) with other types of syndromes. CONCLUSIONS: The main TCM syndrome types of male infertility in Chengdu area include kidney-yang deficiency, kidney-yang deficiency with liver-qi stagnation, and kidney-yang deficiency with spleen-asthenia and excessive dampness. The distribution and influencing factors of the syndrome types need to be further explored and clarified by more large-sample and high-quality studies.

[Correlation of reproductive hormone levels and seminal plasma oxidative stress with semen quality in obese males].

OBJECTIVE: To investigate the correlation of the levels of reproductive hormones and oxidative stress in the seminal plasma with semen parameters in obese males. METHODS: Based on the body mass index (BMI), we divided 138 infertile men into three groups: normal (BMI <24 kg/m2, n = 48), overweight (24 kg/m2≤BMI<28 kg/m2, n = 47), and obesity (BMI ≥28 kg/m2, n = 43). We determined the concentrations of follicle-stimulating hormone (FSH), luteotropic hormone (LH), prolactin (PRL), testosterone (T) and estradiol (E2) in the serum by electrochemiluminescence and measured the levels of superoxide dismutase (SOD), glutathione-S-transferases (GSTs), reactive oxygen species (ROS) and malondialdehyde (MDA) in the seminal plasma by ELISA, compared the above indexes among the three groups, and analyzed their correlation with the semen volume, sperm concentration, total sperm count, and percentage of progressively motile sperm (PMS). RESULTS: The semen volume was significantly lower in the obesity than in the normal group (ï¼»2.63 ± 0.74ï¼½ vs ï¼»3.37 ± 1.00ï¼½ ml, P < 0.05), and so was the percentage of PMS in the overweight and even lower in the obesity than in the normal group (ï¼»47.91 ± 12.89ï¼½ and ï¼»41.27 ± 15.77ï¼½ vs ï¼»54.04 ± 13.29ï¼½%, P < 0.05). Compared with the normal group, both the overweight and obesity groups showed markedly decreased levels of serum T (ï¼»4.83 ± 1.42ï¼½ vs ï¼»3.71 ± 1.22ï¼½ and ï¼»3.49 ± 1.12ï¼½ ng/ml, P<0.05), T/LH ratio (1.53 ± 0.57 vs 1.19 ± 0.54 and 0.97 ± 0.51, P<0.05), SOD (ï¼»112.05 ± 10.54ï¼½ vs ï¼»105.85 ± 6.93ï¼½ and ï¼»99.33 ± 8.39ï¼½ U/ml, P<0.05), and GSTs (ï¼»31.75±6.03ï¼½ vs ï¼»29.54±5.78ï¼½ and ï¼»29.02±4.52ï¼½ U/L, P<0.05), but remarkably increased seminal plasma ROS (ï¼»549.93±82.41ï¼½ vs ï¼»620.61±96.13ï¼½ and ï¼»701.47±110.60ï¼½ IU/ml, P<0.05) and MDA (ï¼»7.46 ± 2.13ï¼½ vs ï¼»8.72 ± 1.89ï¼½ and ï¼»10.47 ± 2.10ï¼½ nmol/L, P<0.05). BMI was correlated positively with ROS and MDA, but negatively with the semen volume, PMS, T, T/LH, SOD and GSTs (P<0.05); LH negatively with sperm concentration, total sperm count and GSTs (P<0.05); PRL negatively GSTs (P<0.05); E2 positively with SOD (P<0.05); T positively with SOD (P<0.05) but negatively with MDA (P<0.05); T/LH positively with PMS and SOD (P<0.05) but negatively with ROS and MDA (P<0.05); SOD positively with semen volume, PMS and GSTs (P<0.05) but negatively with ROS and MDA (P<0.05); GSTs negatively with sperm concentration; total sperm count and MDA (P<0.05); ROS positively with MDA (P<0.01) but negatively with PMS (P<0.05); and MDA negatively with semen volume (P<0.05). Multivariate logistic regression analysis showed that the independent factors influencing the semen volume were BMI and GSTs, those influencing the total sperm count were BMI and T, and those influencing PMS were BMI and MDA. CONCLUSIONS: Increased BMI induces changes in the levels of male reproductive hormones and seminal plasma oxidative stress and affects semen quality, which may be associated with male infertility.

Current approaches for the treatment of male infertility with stem cell therapy.

Infertility is the inability to conceive after having regular unprotected sexual intercourse for more than 12 months and it can be caused by reproductive problems of men, women, or both, but sometimes the cause of infertility is unknown. At least 30 million men around the world suffer from infertility, most of whom are in Africa and Eastern Europe. Since infertility problems can have devastating emotional effects on couples as well as negative effects on the society, treatment of infertility is very important, as a result of which many studies have been carried out in this field and many treatments have been proposed. Recently, due to several advances in infertility treatments, more than 80% of infertile couples now have children. Among these treatments, stem cells as undifferentiated cells and because of their self-renewing and high differentiating potential have been considered by researchers. This review explained different types of male infertility and various therapies, with emphasis on stem cells therapy.

Validity of Claims Data for the Identification of Male Infertility.

PURPOSE OF REVIEW: We sought to determine whether infertile men can accurately be identified within a large insurance claims database to validate its use for reproductive health research. RECENT FINDINGS: Prior literature suggests that men coded for infertility are at higher risk for chronic disease though it was previously unclear if these diagnostic codes correlated with true infertility. We found that the specificity of one International Classification of Disease (9th edition) code in predicting abnormal semen parameters was 92.4%, rising to 99.8% if a patient had three different codes for infertility. The positive predictive value was as high as 85%. The use of claims data for male infertility research has been rapidly progressing due to its high power and feasibility. The high specificity of ICD codes for men with abnormal semen parameters is reassuring and validates prior studies as well as future investigation into men's health.

Estandarización en la técnica de pretratamiento y tinción para la realización de la morfología espermática humana automatizada tipo asma (assisted sperm morphometry analysis) / Standardization of pretreatment and staining technique to perform automatic human sperm morphology with asma (assisted sperm morphometry analyses)

Objetivo. Estandarización de la técnica de pretratamiento y tinción para la automatización de la morfología espermática con metodología tipo computer-assisted sperm morphometry analysis, con el analizador de sémenes SCA 5.4 (Sperm Class Analyzer, Microptic). Material y métodos. La automatización de la morfología se ha realizado con el analizador de semen SCA 5.4 (Microptic S.L., Barcelona, España). El método de tinción ha sido una modificación del equipo Hemacolor (Merck). Se procesan entre 69 y 125 sémenes, los cuales han sido muestras escogidas aleatoriamente de nuestro laboratorio. Resultados. El pretratamiento de la muestra de semen escogido, debido a los resultados obtenidos, fue una centrifugación suave durante 5 min a 300g, se descarta el plasma seminal y se homogeneiza suavemente el sedimento con 0,2ml del propio plasma seminal. Este pretratamiento ya se comprobó que no artefactaba los espermatozoides. La tinción que se ha escogido es el kit Hemacolor (Merck), pero con modificaciones. Se tampona el fijador con buffer fosfato pH 7,2 al 10%, se reduce los tiempos aconsejados por el fabricante a fijación durante 5seg, tinción con eosina 30 seg y tinción con azur 2 seg. Finalmente se lava 30 seg con tampón fosfato pH 7,2, como indica el fabricante. Tras dicho pretratamiento y tinción estandarizadas se hallan los coeficientes de variación del pretratamiento y valores de referencia para nuestra metodología. Conclusiones. La automatización de la morfología espermática reduce los coeficientes de variación de la determinación, aumentando su fiabilidad técnica y eliminando la subjetividad que conlleva un análisis manual. Esta estandarización constituye el primer paso para el estudio del valor diagnóstico de la morfología avanzada en la infertilidad y en enfermedades urológicas (AU)

Objective. Standardization of pretreatment and staining technique to realize morphological semen evaluation with computer-assisted sperm morphometry analysis system, with semen system analyzer SCA 5.4 (Sperm Class Analyzer, Microptic). Material and methods. Morphological analysis was performed with semen system analyzer SCA 5.4 (Microptic SL, Barcelona, Spain). The staining method was a modification of Hemacolor kit (Merck). Between 60 and 125 semen samples were chosen randomly from our laboratory. Results. Pretreatment of semen samples was a centrifugation for 5minutes at 300g, seminal plasma was rejected and the pellet was homogenized with .2mL of the seminal plasma itself. Not change in sperm morphology have been found with this pretreatment. Staining was performed with Hemacolor kit (Merck) but with some modifications. Fixer has been buffered with phosphate buffer pH 7.2 at 10%, time recommended by the manufacturer has been reduced. Fixation for 5 seconds, 30 seconds with Eosin staining and 2 seconds with staining Azur. Finally it was washed for 30 seconds with pH 7.2 phosphate buffer, as indicated by the manufacturer. After the pretreatment and staining we have got reference values for our methodology. Conclusions. An automation methodology to perform sperm morphology reduces coefficient variations of determination thereby we can increase its technical reliability and remove the subjectivity of the manual analysis. This standardization can be the first step to study the diagnostic value of advanced morphology in infertility and urological diseases (AU)

PATTERNS OF SEMINAL FLUID ANALYSIS IN MALE PARTNERS OF INFERTILE COUPLES ATTENDING GYNAECOLOGY CLINIC AT FEDERAL MEDICAL CENTRE, ABEOKUTA.

BACKGROUND: Infertility affects 19% of the general population I, and this constitutes a reproductive health concern for the affected couples. The contribution of male factors as cause of infertility is increasingly being noted in the recent times and has become a source of concern to the affected couples with its attendant social and psychological effects and with the potential of threatening relationships. OBJECTIVES: To assess the seminal fluid analysis parameters in male partners of infertile couples presenting at gynaecological clinic of Federal Medical Centre, Abeokuta and to determine the patterns of seminal fluid abnormalities in the seminalysis results. METHODOLOGY: The study is a 3 year retrospective review of seminal fluid analysis results of male partners in infertility cases at the Federal Medical Centre Abeokuta. Analysis was done using the WHO laboratory manual for the examination and processing of human semen revised fifth edition. RESULTS: During this study period, a total of 214 semen samples were analysed for semen quality over a 3 year period. Sixty four (30%) of the men had normal semen parameters, while one hundred and fifty (70%) had abnormal semen parameters. The abnormal semen parameters consists of low volume (12.6%), prolonged liquefaction time (9.8%), oligospermia (28%), azoospermia (8%), asthenozoospermia (25%), teratozoospermia (9%), combined defects of oligo-asthenozoospermia (23.8%), oligo-teratozoospermia (9.8%), asthenoteratozoospermia (12.60%) and oligoasthenoteratozoospermia (11.20%). CONCLUSION: This study has confirmed that male factor infertility remains a significant contributor to infertility in our environment. Efforts should be made in enlightening men on the common aetiologies of abnormal semen and options of treatment of likely causes.

Total motile sperm count: a better indicator for the severity of male factor infertility than the WHO sperm classification system.

STUDY QUESTION: Does the prewash total motile sperm count (TMSC) have a better predictive value for spontaneous ongoing pregnancy (SOP) than the World Health Organization (WHO) classification system? SUMMARY ANSWER: The prewash TMSC shows a better correlation with the spontaneous ongoing pregnancy rate (SOPR) than the WHO 2010 classification system. WHAT IS KNOWN ALREADY: According to the WHO classification system, an abnormal semen analysis can be diagnosed as oligozoospermia, astenozoospermia, teratozoospermia or combinations of these and azoospermia. This classification is based on the fifth percentile cut-off values of a cohort of 1953 men with proven fertility. Although this classification suggests accuracy, the relevance for the prognosis of an infertile couple and the choice of treatment is questionable. The TMSC is obtained by multiplying the sample volume by the density and the percentage of A and B motility spermatozoa. STUDY DESIGN, SIZE, DURATION: We analyzed data from a longitudinal cohort study among unselected infertile couples who were referred to three Dutch hospitals between January 2002 and December 2006. Of the total cohort of 2476 infertile couples, only the couples with either male infertility as a single diagnosis or unexplained infertility were included (n = 1177) with a follow-up period of 3 years. PARTICIPANTS/MATERIALS, SETTING, METHODS: In all couples a semen analysis was performed. Based on the best semen analysis if more tests were performed, couples were grouped according to the WHO classification system and the TMSC range, as described in the Dutch national guidelines for male infertility. The primary outcome measure was the SOPR, which occurred before, during or after treatments, including expectant management, intrauterine insemination, in vitro fertilization or intracytoplasmic sperm injection. After adjustment for the confounding factors (female and male age, duration and type of infertility and result of the postcoital test) the odd ratios (ORs) for risk of SOP for each WHO and TMSC group were calculated. The couples with unexplained infertility were used as reference. MAIN RESULTS AND THE ROLE OF CHANCE: A total of 514 couples did and 663 couples did not achieve a SOP. All WHO groups have a lower SOPR compared with the unexplained group (ORs varying from 0.136 to 0.397). Comparing the couples within the abnormal WHO groups, there are no significant differences in SOPR, except when oligoasthenoteratozoospermia is compared with asthenozoospermia [OR 0.501 (95% CI 0.311-0.809)] and teratozoospermia [OR 0.499 (95% CI: 0.252-0.988)], and oligoasthenozoospermia is compared with asthenozoospermia [OR 0.572 (95% CI: 0.373-0.877)]. All TMSC groups have a significantly lower SOPR compared with the unexplained group (ORs varying from 0.171 to 0.461). Couples with a TMSC of <1 × 10(6) and 1-5 × 10(6) have significantly lower SOPR compared with couples with a TMSC of 5-10 × 10(6) [respectively, OR 0.371 (95% CI: 0.215-0.64) and OR 0.505 (95% CI: 0.307-0.832)]. LIMITATIONS, REASON FOR CAUTION: To include all SOPs during the follow-up period of 3 years, couples were not censured at the start of treatment. WIDER IMPLICATIONS OF THE FINDINGS: Roughly, three prognostic groups can be discerned: couples with a TMSC <5, couples with a TMSC between 5 and 20 and couples with a TMSC of more than 20 × 10(6) spermatozoa. We suggest using TMSC as the method of choice to express severity of male infertility. STUDY FUNDING/COMPETING INTERESTS: None.

Light microscopy morphological characteristics of the sperm flagellum may be related to axonemal abnormalities.

Although electron microscopy provides a detailed analysis of ultrastructural abnormalities, this technique is not available in all laboratories. We sought to determine whether certain characteristics of the flagellum as assessed by light microscopy were related to axonemal abnormalities. Forty-one patients with an absence of outer dynein arms (type I), a lack of a central complex (type III) and an absence of peripheral doublets (type IV) were studied. Sperm morphology was scored according to David's modified classification. Flagella with an irregular thickness were classified as being of normal length, short or broken. There were correlations between missing outer dynein arms and abnormal, short or coiled flagellum. Type III patients showed the highest flagellar defects (a short (P = 0.0027) or an absent flagellum (P = 0.011)). Just over 68% of the irregular flagella were short in Type III patients, whereas this value was only 34.5% in type I and 26.4% in type IV (P = 0.002). There was a negative correlation between misassembly and spermatozoa of irregular flagella (r = -0.79; P = 0.019). It is concluded that light microscopy analysis of flagellum abnormalities may help provide a correct diagnosis, identify sperm abnormalities with fertility potentials and outcomes in assisted reproduction technologies and assess the genetic risk.

[IL-6 and sICAM-1 in seminal plasma relate to male immune infertility].

OBJECTIVE: To detect the expressions of interleukin-6 (IL-6) and soluble intercellular adhesion molecule-1 (sI- CAM-1) in the seminal plasma of infertile men and explore the role of inflammatory cytokines in male immune infertility. METHODS: Based on the results of the sperm-bound antibody immunobead test, 123 males with clinically suspected infertility were divided into an immune infertility group (n = 41), other infertility group A (n = 37), and other infertility group B (n = 45). The immune infertility patients were further subdivided into a leukocyte-positive and a leukocyte-negative group according to the results of leukocyte peroxidase staining. The control group included 31 males confirmed to be fertilein the clinic. Statistical analyses were conducted on the differences in inflammatory cytokines expressions and main parameters in the seminal plasma among different groups. The seminal liquefaction time was measured by visual and microscopic observation, sperm concentration and motility detected using the computer-assisted sperm analysis system, sperm viability determined by hypotonic swelling assay, and the expression levels of IL-6 and sICAM-1 meas- ured by ELISA. RESULTS: The infertility groups showed significantly lowers perm viability (P < 0.05) and progressive motility (P < 0.01) than the fertile control, but no remarkable differences from the latter in sperm concentration (P > 0.05) and semen liquefaction time (P > 0.05). No statistically significant differences were observed in seminal parameters between the immune infertility group and other infertility groups (P > 0.05). The IL-6 and sICAM-1 levels in the seminal plasma were extremely significantly higher in the im- mune infertility group ([37.92 ± 17.01] ng/L and [89.15 ± 41.82] ng/ml), other infertility group A ([22.23 ± 13.77] ng/L and [67.81 ± 33.24] ng/ml), and other infertility group B ([18.75 ± 14.32] ng/L and [53.25 ± 27.09] ng/ml) than in the normal control group ([9.47 ± 5.76] ng/L and [19.46 ± 9.77] ng/ml) (P <0.01) , with remarkable differences between the immune infertility group and the other two infertility groups (P < 0.05). The leukocyte-positive patients showed significantly increased levels of IL-6 ([49.25 ± 21.46] ng/L) and sICAM-1 ([104.36 ± 46.41] ng/ml) as compared with the leukocyte-negative ones ([31.38 ± 15.54] ng/Land [80.38 ± 35.52] ng/ml) (both P < 0.05). CONCLUSION: IL-6 and sICAM-1 in the seminalplasma are involved in male immune infertility.

Protamine-1 and -2 polymorphisms and gene expression in male infertility: an Italian study.

BACKGROUND: Correct histone/protamine replacement is an important stage in chromatin condensation during spermiogenesis in humans. There are two types of protamines: protamine 1 (P1) and the protamine 2 family (P2, P3, and P4), coded by the genes PRM1 and PRM2. AIM: We analyze the sequences and gene expression of PRM1 and PRM2 and their relationship with defective spermatogenesis. MATERIALS AND METHODS: Sequence analysis was carried out on 163 patients attending our laboratory for analysis of seminal fluid. Patients were divided into three groups: normozoospermic (53), teratozoospermic (60), and azoospermic (50). Gene expression was analyzed in seven patients with azoospermia and one with cryptozoospermia. RESULTS: Seven single nuclotide polymorphisms (SNP) were identified: G54A, G102T and C230A for PRM1, and C246T, G288C, G298C and C373A for PRM2. For C230A, the CA genotype was present in 38% of teratozoospermic vs 55% of normozoospermic and 64% of azoospermic patients; for C373A, CA was found in 37% of teratozoospermic vs 47% of normozoospermic and 64% of azoospermic patients. In contrast, for G298C, GC was more common in the teratozoospermic (63%) than in the normozoospermic (49%) or azoospermic (48%) groups. These differences could suggest a greater susceptibility of these patients to abnormal sperm morphology. In five patients the levels of transcripts were reduced with respect to the control. CONCLUSION: These data suggest that premeiotic arrest is associated with extremely reduced protamine expression. New studies of both PRM1 and PRM2 and their mRNA expression could help us better understand the molecular mechanisms underlying the protamine transcription and translation processes.

Patient and hospital characteristics associated with variation in guideline adherence in intrauterine insemination care.

OBJECTIVE: To assess the association of patient and hospital characteristics with adherence to guidelines for intrauterine insemination (IUI) care. DESIGN: Retrospective cohort study using multilevel regression analysis. Characteristics studied at the patient level were female age, type and duration of subfertility, diagnosis and number of started IUI cycles. At the hospital level, the characteristics studied were hospital size, teaching hospital, in vitro fertilization (IVF) licence and number of physicians involved in the IUI programme. Data were obtained from medical records and questionnaires for gynaecologists. SETTING AND PARTICIPANTS: Five hundred and fifty-eight subfertile couples who underwent IUI treatment at 10 Dutch hospitals. MAIN OUTCOME MEASURES: Adherence to systematically developed guideline-based performance indicators describing 20 processes of IUI care. RESULTS: A total of 558 couples who started 2,334 IUI cycles participated. Guideline adherence in IUI care was often substandard and varied considerably between hospitals. Variation in guideline adherence in IUI care was associated with the patient characteristics 'diagnosis' and 'female age'. Only adherence to the guideline recommendation regarding 'screening for tubal occlusion' was associated with hospital characteristics ('hospital size' and 'IVF licence'). Large explained variances up to 39% were found for the different models. CONCLUSIONS: A number of patient and hospital characteristics were associated with variation in guideline adherence in IUI care, particularly the patient characteristics 'diagnosis' and 'female age'. The identification of different subgroups in the patient population and different types of hospitals with regard to the extent of guideline adherence in IUI care is important for the tailoring of interventions to improve IUI care.

A prospective randomized noninferiority study comparing recombinant FSH and highly purified menotropin in intrauterine insemination cycles in couples with unexplained infertility and/or mild-moderate male factor.

OBJECTIVE: To demonstrate the noninferiority of highly purified menotropin (HP-hMG) compared with recombinant FSH (rFSH) regarding clinical pregnancy rate (PR) in intrauterine insemination (IUI) cycles. DESIGN: Prospective randomized noninferiority trial. SETTING: Unit of physiopathology of human reproduction, university hospital. PATIENT(S): Five hundred twenty-three patients with unexplained infertility or mild male infertility undergoing controlled ovarian hyperstimulation for IUI. INTERVENTION(S): Patients were randomized for treatment with rFSH (262 patients) or HP-hMG (261 patients). Insemination was performed 34-36 hours after hCG injection. MAIN OUTCOME MEASURE(S): The primary outcome was clinical pregnancy rate (PR). The secondary outcome was the number of interrupted cycles for high risk of ovarian hyperstimulation syndrome (OHSS) and multiple pregnancy. RESULT(S): The clinical PR was 19.7% (95% confidence interval [CI] 15.3%-25.1%) in the HP-hMG group and 21.4% (95% CI 16.9%-26.8%) in the rFSH group [absolute difference -1.7% (95% CI -8.6%-5.2%)]; therefore, the noninferiority was demonstrated. The number of interrupted cycles for OHSS risk and multiple pregnancy was significantLy higher in the rFSH group, 8.4% (95% CI 5.6%-12.4%) than in the HP-hMG group 1.2% (95% CI 0.4%-3.3%) [absolute difference -7.27% (95% CI -11.3 to -3.7)]. CONCLUSION(S): HP-hMG is not inferior compared with rFSH regarding clinical PR.

Development of dimethandrolone 17beta-undecanoate (DMAU) as an oral male hormonal contraceptive: induction of infertility and recovery of fertility in adult male rabbits.

Dimethandrolone undecanoate (DMAU: 7α,11ß-dimethyl-19-nortestosterone 17ß-undecanoate) is a potent orally active androgen with progestational activity that is in development for therapeutic uses in men. We hypothesized that because of its dual activity, DMAU might have potential as a single-agent oral hormonal contraceptive. To test this possibility, adult male rabbits (5/group) of proven fertility were treated orally with vehicle or DMAU at 1.0, 2.5, 5.0, or 10.0 mg/kg/d for 12 or 13 weeks. Semen and blood samples were collected every other week through week 30. Sperm were decreased (P < .05) in semen samples from DMAU-treated rabbits at 2.5 and 5.0 mg/kg/d at weeks 12, 14, 16, 18, and 20 compared to week 0 (prior to treatment). The percentage of forward progressive motile sperm in those rabbits that still had measurable sperm was also reduced by DMAU treatment at 2.5 mg/kg/d at weeks 14, 16, 18, and 20 and at 5.0 mg/kg/d at week 18 (P < .05). At 1.0 mg/kg/d only 1 rabbit had reduced sperm numbers and motility. A mating trial was performed at week 15. The number of bred males that were fertile was 4 of 4 in the vehicle-treated group and 4 of 5, 0 of 4, and 2 of 5 in the 1.0, 2.5, and 5.0 mg/kg/d DMAU treatment groups. By week 22, sperm numbers and forward progressive motility increased, and they returned to pretreatment levels in all DMAU-treated rabbits by week 30. All bred males were fertile at week 31. Serum levels of testosterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) were significantly suppressed in DMAU (1.0, 2.5, or 5.0 mg/kg/d)-treated rabbits during the 12-week dosing interval, but were comparable to pretreatment levels after cessation of dosing. These data indicate that DMAU suppressed the hypothalamic-pituitary-gonadal axis, resulting in severe oligospermia in the majority of rabbits in the 2.5 and 5.0 mg/kg/d dosing groups. Infertility was observed when sperm numbers decreased to about 10% of pretreatment levels. In rabbits dosed with DMAU at 10.0 mg/kg/d, no effect on sperm numbers or motility was observed by week 12. Dosing continued for another week, and the rabbits underwent a gross necropsy on week 13 with removal of testes and epididymides for histology and preparation of testicular cytosol. Serum testosterone, FSH, and LH levels were considerably suppressed in these rabbits as in the lower-dose groups. The lack of oligospermia in the 10.0 mg/kg group as well as in the 2 fertile males in the 5.0 mg/kg group may have been due to high intratesticular levels of 7α,11ß-dimethyl-19-nortestosterone, the active metabolite of DMAU. Hence, as observed previously for testosterone, DMAU has a biphasic effect on spermatogenesis. Collectively, these data indicate that DMAU has the potential to be an orally active single-agent male hormonal contraceptive at an appropriate dose level and should be tested for contraceptive efficacy in nonhuman primates.

Alterations in sperm DNA methylation patterns at imprinted loci in two classes of infertility.

OBJECTIVE: To evaluate the associations between proper protamine incorporation and DNA methylation at imprinted loci. DESIGN: Experimental research study. SETTING: Research laboratory. PATIENT(S): Three populations were tested-abnormal protamine patients, oligozoospermic patients, and fertile donors. INTERVENTION(S): The CpG methylation patterns were examined at seven imprinted loci sequenced: LIT1, MEST, SNRPN, PLAGL1, PEG3, H19, and IGF2. MAIN OUTCOME MEASURE(S): The DNA methylation patterns were analyzed using bisulfite sequencing. The percentage of methylation was compared between fertile and infertile patients displaying abnormal protamination. RESULT(S): At six of the seven imprinted genes, the overall DNA methylation patterns at their respective differentially methylated regions were significantly altered in both infertile patient populations. When comparing the severity of methylation alterations among infertile patients, the oligozoospermic patients were significantly affected at mesoderm-specific transcript (MEST), whereas abnormal protamine patients were affected at KCNQ1, overlapping transcript 1 (LIT1), and at small nuclear ribonucleoprotein polypeptide N (SNRPN). CONCLUSION(S): Patients with male factor infertility had significantly increased methylation alteration at six of seven imprinted loci tested, with differences in significance observed between oligozoospermic and abnormal protamine patients. This could suggest that risk of transmission of epigenetic alterations may be different with diagnoses. However, this study does not provide a causal link for epigenetic inheritance of imprinting diseases, but does show significant association between male factor infertility and alterations in sperm DNA methylation at imprinted loci.

Changes in affect and state anxiety across an in vitro fertilization/intracytoplasmic sperm injection cycle.

OBJECTIVE: To identify pattern of change in average positive affect (PA), negative affect (NA), and state anxiety (St ANX) across three biological end points of an IVF/intracytoplasmic sperm injection (ICSI) procedure and to examine whether the pattern varied across sociodemographic and biomedical subgroups. DESIGN: Longitudinal follow-up study of PA, NA, and St ANX at three different time points: before start of study, before ovum pick-up (OPU), and before embryo transfer. SETTING: Three infertility centers in northern India. PATIENT(S): Baseline data were obtained from a consecutive sample of 85 women. However, final analysis was done on data obtained from 74 women who reached the embryo transfer stage and completed the questionnaires at both OPU and embryo transfer. INTERVENTION(S): The PA, NA, and St ANX scores. MAIN OUTCOME MEASURE(S): Change in PA, NA, and St ANX scores at three stages of the treatment: baseline (T(0)), before OPU (T(1)), before embryo transfer (T(2)). RESULT(S): The PA scores before OPU and embryo transfer were significantly lower than those at baseline. The mean NA and St ANX scores before OPU and embryo transfer were significantly higher than baseline scores. Furthermore, mean NA before embryo transfer was significantly higher than mean NA before OPU. The PA and St ANX scores showed statistically insignificance within cycle variations. Furthermore, the mean PA and St ANX for a subgroup of women who reported more than moderate level of burden were less variable. CONCLUSION(S): The OPU and embryo transfer stages are more stressful than the baseline stage for most women across various sociodemographic and biomedical subgroups. Women with more than a moderate level of financial burden were relatively more stable.

Seminal plasma total antioxidant capacity and semen parameters in patients with varicocele.

Total antioxidant capacity (TAC) was evaluated in the seminal plasma of infertile patients with varicocele in relation to their semen parameters. The study recruited 60 patients affected by varicocele and 10 fertile non-varicocele subjects as controls. Controls had normal semen parameters and proven fertility. On the basis of semen parameters, patients with varicocele were grouped into normozoospermic (n = 12), asthenozoospermic (n = 8), oligoasthenozoospermic (n = 40). The group with oligosthenozoospermia was divided into mild (<20 x 10(6)/ml; > or =15 x 10(6)/ml), moderate (<15 x 10(6)/ml; > or =5 x 10(6)/ml), and severe (<5 x 10(6)/ml), based on sperm count. Antioxidant activity was measured in seminal plasma and peripheral blood using the free oxygen radicals defence test. No significant differences were observed in peripheral blood TAC concentrations between controls and groups. In patients with varicocele and moderate oligoasthenozoospermia or severe oligoasthenozoospermia, seminal plasma TAC concentrations were significantly lower (P < 0.05) than in controls and normozoospermic patients with varicocele. Moreover, in patients with severe oligosthenozoospermia, seminal plasma TAC concentrations were also significantly lower (P < 0.05) than in asthenozoozpermic patients with varicocele. In all subjects, concentrations of TAC showed a positive correlation with sperm concentration (r = 0.93, P < 0.05) and motility (r = 0.92, P < 0.05).

Y chromosome microdeletions in Tunisian infertile males.

AIM: To determine frequency of Y microdeletions in azoospermic and oligospermic Tunisian infertile males. METHODS: A Sample of 146 Tunisian infertile males with a low sperm count (<5 x 10(6) sperms per mililiter) and normal karyotype was screened for Y chromosome microdeletions. 76 men were azoospermic and 70 men were oligospermic. Genomic DNA was isolated from blood and multiplex PCR was carried out with a set of 20 AZFa, AZFb and AZFc STS markers to detect the microdeletions as recommended by the European Academy of Andrology. RESULTS: In 10/146 (6.85%) subjects AZF deletions were observed. Of these ten males with microdeletions, 9/10 subjects were azoospermic (90%), 1/10 was oligospermic (10%). Frequency of microdeletions in azoospermic men was 9/76 (11.84%). None of the patients showed isolated microdeletion in the AZFa region, but one azoospermic man had deletion in the AZFb region. Eight azoospermic patients and one oligospremic man have AZFc microdeletions. AZFc and AZFb were deleted in three azoospermic patients. AZFc, AZFb and AZFa were deleted in three azoospermic patients We estimate the sensitivity of the test comprising six STS in our sample to be 90%. CONCLUSION: The incidence of Yq microdeletions in the study population of infertile Tunisian men falls within the range published in other countries. We suggest to analyze 9STS in the first step to detect efficiently Y microdeletions in our population.