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1.
Khirurgiia (Mosk) ; (10): 44-48, 2020.
Artigo em Russo | MEDLINE | ID: mdl-33047585

RESUMO

OBJECTIVE: To reveal the association of tension-free inguinal hernia repair and pathospermia in fertile men. MATERIAL AND METHODS: We have retrospectively analyzed medical records of 512 men who appealed to andrologist with complaints of the absence of pregnancy in wife in 2018. We evaluated duration and features of infertility, presence/absence of previous inguinal hernia repair, spermogram data (according to WHO criteria, 2010) in all patients. RESULTS: Duration of infertility in men after inguinal hernia repair persists for 4.2±2.1 years. Right-sided hernia repair was performed in 36 (48.6%) patients, left-side - 23 (31%), bilateral repair - 15 (20.2%) patients. Men with impaired sperm motility prevailed among patients after right-sided inguinal hernia repair (17 (47.2%) people). Left-sided hernia repair was followed by asthenozoospermia in 8 (34.7%) cases, bilateral hernia repair - in 3 (20%) cases. The most severe abnormalities in semen analysis (azoospemia) develop after bilateral hernia repair. CONCLUSION: Inguinal tension-free hernia repair is a risk factor for male infertility in 14.4% of cases. It is very important to examine a man in case of infertile marriage. Previous surgical interventions including inguinal hernia repair should be considered.


Assuntos
Hérnia Inguinal/cirurgia , Herniorrafia/efeitos adversos , Infertilidade Masculina/etiologia , Astenozoospermia/diagnóstico , Astenozoospermia/etiologia , Azoospermia/diagnóstico , Azoospermia/etiologia , Herniorrafia/métodos , Humanos , Infertilidade Masculina/diagnóstico , Masculino , Estudos Retrospectivos , Fatores de Risco , Análise do Sêmen
2.
J Urol ; 204(6): 1312-1317, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32799727

RESUMO

PURPOSE: In 2012 the American Urological Association published vasectomy guidelines to promote best practices, including when to obtain post-vasectomy semen analyses. In this study we assessed practice patterns of post-vasectomy semen analysis since this guideline publication. MATERIALS AND METHODS: We retrospectively analyzed a database of men who underwent post-vasectomy semen analysis between 2013 and 2017. Vasectomies were performed by urologist and nonurologist providers in academic and community settings. RESULTS: A total of 4,827 men underwent post-vasectomy semen analysis with 22.3% undergoing 1 or more repeat analyses. On initial analysis 58.2% were azoospermic, 28.3% had less than 100,000/ml rare nonmotile sperm, 8.7% had greater than 100,000/ml nonmotile sperm and 4.8% had motile sperm. The rate of repeat post-vasectomy semen analysis decreased from 30.7% in 2013 to 18.6% in 2016. Overall 72% of repeat post-vasectomy semen analyses were performed for patients with azoospermia or rare nonmotile sperm on initial post-vasectomy semen analysis. Of the 421 men with greater than 100,000/ml nonmotile sperm, 61.3% did not obtain a repeat analysis. Among cases of repeat analysis after initially having greater than 100,000/ml nonmotile sperm, 67.5% were downgraded to rare nonmotile sperm or azoospermia, 32.5% had a persistent count greater than 100,000/ml nonmotile sperm and none developed motile sperm. CONCLUSIONS: The rate of repeat post-vasectomy semen analysis is decreasing, likely highlighting a decrease in unnecessary testing. However, there is ongoing discordance between vasectomy guidelines and practice patterns, with 72% of repeat post-vasectomy semen analyses obtained unnecessarily based on guideline recommendations. Interestingly, no men with greater than 100,000/ml nonmotile sperm went on to have motile sperm on repeat post-vasectomy semen analysis. Further provider education is warranted and subsequent studies may allow for guideline modification wherein all nonmotile sperm are characterized similarly.


Assuntos
Azoospermia/diagnóstico , Fidelidade a Diretrizes/estatística & dados numéricos , Padrões de Prática Médica/estatística & dados numéricos , Análise do Sêmen/estatística & dados numéricos , Vasectomia , Adulto , Azoospermia/etiologia , Humanos , Masculino , Período Pós-Operatório , Guias de Prática Clínica como Assunto , Padrões de Prática Médica/normas , Estudos Retrospectivos , Análise do Sêmen/normas , Sociedades Médicas/normas , Fatores de Tempo , Estados Unidos , Urologia/normas
3.
Fertil Steril ; 113(4): 774-780.e3, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32228879

RESUMO

OBJECTIVE: To assess the timing of patency and late failure (secondary azoospermia) after vasovasostomy (VV) using standardized kinetics definitions. DESIGN: Retrospective cohort study. SETTING: University-affiliated hospital. PATIENT(S): Patients with obstructive azoospermia. INTERVENTION(S): Vasovasostomy. MAIN OUTCOME MEASURE(S): Univariate and multivariate logistic regression assessed predictors of patency and late failure. Patency was defined as any sperm return to the ejaculate; and >2 million total motile sperm (TMS) in ejaculate. Late failure after VV was defined as azoospermia; or <2 million TMS in ejaculate. RESULT(S): 429 men underwent VV, with median follow up of 242 days. Mean time to patency was 3.25 months versus 5.29 months in the "any sperm" versus ">2 million TMS" groups. Finding sperm intraoperatively during VV significantly improved patency rates in multivariable analysis (odds ratio [OR] 4.22). This association was further boosted when sperm was found bilaterally (OR 6.70). Late failure rate (azoospermia) was 10.6% at mean time of 14.1 months and 23% for <2 million, at mean time of 15.7 months. When assessing predictors of late failure, intraoperative motile sperm bilaterally was a statistically significant protective factor on multivariate analysis (hazard ratio 0.22). CONCLUSION(S): Vasovasostomy remains highly efficacious in treating obstructive azoospermia. Young patients, shorter obstructive intervals, and sperm identified intraoperatively predict improved outcomes. Clinicians can expect VV patency in 3 months and late failure within the first 2 years after surgery. However, patency rates, late failure rates, and kinetics vary by definition.


Assuntos
Azoospermia/diagnóstico , Ducto Deferente/cirurgia , Vasovasostomia/métodos , Adulto , Azoospermia/fisiopatologia , Estudos de Coortes , Seguimentos , Humanos , Cinética , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Estudos Retrospectivos , Motilidade Espermática/fisiologia , Resultado do Tratamento , Ducto Deferente/fisiopatologia , Vasovasostomia/tendências
4.
Urol Clin North Am ; 47(2): 147-155, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32272986

RESUMO

For men with obstructive azoospermia, several surgical sperm retrieval techniques can facilitate conception with assisted reproductive technology. The evolution of both percutaneous and open approaches to sperm retrieval has been affected by technological innovations, including the surgical microscope, in vitro fertilization, and intracytoplasmic sperm injection. Further modifications to these procedures are designed to minimize patient morbidity and increase the quality and quantity of sperm samples. Innovative technologies promise to further ameliorate outcomes by selecting the highest quality sperm. Although various approaches to surgical sperm retrieval are now well established, several advancements in sperm selection and optimization are being developed.


Assuntos
Azoospermia/diagnóstico , Azoospermia/genética , Fertilização In Vitro/métodos , Injeções de Esperma Intracitoplásmicas/métodos , Recuperação Espermática , Fragmentação do DNA , Humanos , Masculino , Análise do Sêmen
5.
Urol Clin North Am ; 47(2): 157-164, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32272987

RESUMO

Guiding a couple with nonobstructive azoospermia requires an integrated approach to care by the urologist and the reproductive endocrinologist. After informing the couple of the implications of the diagnosis, care must be taken to outline the options of parenthood. Most experts agree that sperm retrieval in men can be challenging. This article describes various options of sperm retrieval, historic and contemporary, and highlights the advantages and disadvantages of each. The authors find that using a testicular map can invariably help guide sperm retrieval and overall fertility care. The right approach is one that involves a shared decision with the couple.


Assuntos
Azoospermia/diagnóstico , Azoospermia/genética , Biópsia por Agulha Fina/métodos , Recuperação Espermática , Testículo/patologia , Azoospermia/etiologia , Humanos , Masculino , Microdissecção , Análise do Sêmen , Recuperação Espermática/efeitos adversos , Espermatozoides/patologia
6.
Asian J Androl ; 22(1): 100-105, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31134916

RESUMO

Many studies have shown that microRNAs (miRNAs) play vital roles during the spermatogenesis. However, little is known about the altered miRNA profiles of testicular tissues in nonobstructive azoospermia (NOA). Using microarray technology, the miRNA expression profiles of testicular biopsies from patients with NOA and of normal testicular tissues were determined. Bioinformatics analyses were conducted to predict the enriched biological processes and functions of identified miRNAs. The microarray data were validated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), the results of which were then validated with a larger sample size. Correlations between the miRNA expression levels and clinical characteristics were analyzed. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic ability of miRNAs for azoospermia. Hierarchical clustering showed that 129 miRNAs were significantly differentially expressed between the NOA and control groups. Bioinformatics analysis indicated that the differentially expressed miRNAs were involved in spermatogenesis, cell cycle, and mitotic prometaphase. In the subsequent qRT-PCR assays, the selected miRNA expression levels were consistent with the microarray results, and similar validated results were obtained with a larger sample size. Some clinical characteristics were significantly associated with the expression of certain miRNAs. In particular, we identified a combination of two miRNAs (miR-10b-3p and miR-34b-5p) that could serve as a predictive biomarker of azoospermia. This study provides altered miRNA profiles of testicular biopsies from NOA patients and examines the roles of miRNAs in spermatogenesis. These profiles may be useful for predicting and diagnosing the presence of testicular sperm in individuals with azoospermia.


Assuntos
Azoospermia/genética , MicroRNAs/metabolismo , Espermatogênese/genética , Testículo/metabolismo , Adulto , Azoospermia/diagnóstico , Biópsia , Análise por Conglomerados , Biologia Computacional , Hormônio Foliculoestimulante/metabolismo , Perfilação da Expressão Gênica , Humanos , Hormônio Luteinizante/metabolismo , Masculino , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Testosterona/metabolismo , Análise Serial de Tecidos
7.
Acta Cytol ; 64(3): 216-223, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31550721

RESUMO

BACKGROUND: Infertility is an ever-increasing problem in today's world. It can be due to male or female causes. Azoospermia seen in 5-10% of infertile men is due to obstructive or non-obstructive causes. Traditionally, testicular biopsy is the gold standard for evaluation. Fine-needle aspiration (FNA), however, is minimally invasive, provides qualitative and quantitative information about spermatogenesis, and can aid in assisted reproductive techniques making it a novel technique for the evaluation of male infertility. OBJECTIVE: We aimed to classify different causes of azoospermia into different patterns based upon FNA, and assess the utility of cell indices in classifying cases into different patterns. METHOD: We conducted a prospective and a retrospective study of 42 azoospermic males, confirmed on semen analysis, over a period of 5 years. Patients were subjected to FNA of the testes. Smears were prepared, air-dried, wet-fixed, and then stained with May-Grünwald Giemsa and Papanicolaou stains, respectively. Cells were identified using predetermined morphologic criteria, and various indices were calculated followed by statistical analysis of the observations. RESULTS: The mean age of 40 patients who satisfied the adequacy criteria was 32.75 years (range 22-48 years). Thirty-four patients had primary infertility and 6 had secondary infertility. Of these, 12 had normal spermatogenesis, 8 had hypo-spermatogenesis, 3 had early and 7 had late maturation arrest, 6 had Sertoli cell-only syndrome (SCOS), and there were different results in each testicle in 4 cases. The Sperm Index (SI) was significantly higher in all cases of normal spermatogenesis than in any of the hypo-spermatogenesis cases (p = 0.009). The Sertoli Index (SEI) in cases of hypo-spermatogenesis and maturation arrest was significantly higher than in cases of normal spermatogenesis (p < 0.001). The Sperm-Sertoli Index (SSI) also showed significant differences between cases of hypo-spermatogenesis and normal spermatogenesis (p < 0.001). These indices were useful in categorising patients with azoospermia. CONCLUSION: FNA helps to easily and accurately identify all types of testicular cells without biopsy. SI, SEI, and SSI are powerful cell indices for assessing the extent of spermatogenesis and classifying various causes of azoospermia. Bilateral sampling and multiple aspirations give a better mapping of spermatogenesis within the testes. Testicular FNA can thus play a very important role in the evaluation of male infertility.


Assuntos
Azoospermia/classificação , Azoospermia/diagnóstico , Biópsia por Agulha Fina/métodos , Testículo/cirurgia , Adulto , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
8.
Andrologia ; 52(1): e13453, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31762071

RESUMO

miRNAs (MicroRNAs), known as noncoding and important endogenous factors regulating the expression protein-coding genes, are vital regulators in each biological process. Thus, this study aims to explore the key role of four microRNAs in regulating the spermatogenesis. To conduct this experiment, 55 infertile and fertile men provided the study with the sperm and testicular tissue samples. To study the spermatozoa in terms of the morphology, Diff-Quick was applied. Then, quantitative real-time polymerase chain reaction (RT-PCR) was conducted on samples. Our data indicated that in contrast to the miR-15b, significant increasing of miR-383 and miR-122 occurred in both severe oligoasthenoteratozoospermia (SOAT) and moderate oligoasthenoteratozoospermia (MOAT) compared to normal sperm group (N). In addition, it was observed that miR-15b and miR-122 increased in patients with nonobstructive azoospermia (NOA) compared with obstructive azoospermia (OA) group. Expression levels of target genes including P53, CASPASE-9 and CYCLIN D1 underwent principle changes according to miRNAs expression level. Our finding indicated that miRNAs had essential role in the regulation of spermatogenesis, and their expression altering was associated with sperm abnormalities. Thus, microRNAs can be introduced as useful biomarkers to determine male infertility reasons to choose the effective treatment.


Assuntos
Azoospermia/diagnóstico , Redes Reguladoras de Genes , MicroRNAs/metabolismo , Oligospermia/diagnóstico , Espermatogênese/genética , Adulto , Azoospermia/genética , Biomarcadores/análise , Biomarcadores/metabolismo , Caspase 9/genética , Ciclina D1/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Humanos , Masculino , MicroRNAs/análise , Oligospermia/genética , Espermatozoides/metabolismo , Proteína Supressora de Tumor p53/genética , Adulto Jovem
9.
J Urol ; 203(4): 809-816, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31742474

RESUMO

PURPOSE: We sought to determine whether gene products from spermatogenic cells could be detected in Sertoli cell-only testes. Transcriptome analysis using next generation sequencing was performed using human testicular samples. MATERIALS AND METHODS: This study enrolled 20 men with nonobstructive azoospermia with a histological diagnosis of Sertoli cell-only and no germ cells on in vitro fertilization analysis and 5 with obstructive azoospermia and histologically normal spermatogenesis. Individual differences in the transcriptome of obstructive azoospermia testicular tissues generated on the Illumina® platform were compared as the number of fragments per kilobase of exon per million mapped sequence reads. We confirmed mRNA expression and targeted gene product localization by quantitative reverse transcriptase-polymerase chain reaction and immunohistochemical analysis, respectively. RESULTS: Using next generation sequencing we analyzed 5,666 of the 23,003 screened genes. As representative markers of immature germ cells, UTF1, CD9, DDX4, EPCAM, GFRA1, KIT, LIN28, DMRT, GPR125, UCHL1 and NANOG transcripts were detected in 13 Sertoli cell-only samples (65%). Reverse transcriptase-polymerase chain reaction showed significant mRNA expression of UTF1, CD9, DDX4 and KIT in Sertoli cell-only samples. Immunostaining revealed the localization of DDX4-positive cells, presumably spermatogonia, in 9 Sertoli cell-only samples (45%). These cells lacked proliferative cell nuclear antigen expression. CONCLUSIONS: A number of Sertoli cell-only testes contain immature germ cells (spermatogonia and spermatogenic stem cells) with various gene expression profiles. Sertoli cell-only is a heterogeneous pathophysiology which may be corrected by medical treatment or regenerative technologies.


Assuntos
Azoospermia/patologia , Espermatogênese , Espermatogônias/metabolismo , Testículo/citologia , Adulto , Azoospermia/diagnóstico , Azoospermia/terapia , Biomarcadores/análise , Biomarcadores/metabolismo , Separação Celular/métodos , Humanos , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/metabolismo , RNA-Seq , Células de Sertoli/patologia , Testículo/patologia
10.
Indian J Med Res ; 150(1): 81-86, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31571633

RESUMO

Background & objectives: For improved male contraception, a new polymeric drug molecule - Reversible Inhibition of Sperm under Guidance (RISUG) has been synthesized and has been found to be effective, safe and reversible in various animal species. Phase-I and phase-II clinical trials have confirmed its safety and contraceptive efficacy. The present study was undertaken as a multicentric-limited phase-III clinical trial to test the efficacy and safety of RISUG in human volunteers. Methods: One hundred and thirty nine young males each having at least two children and living with wife were given 120 µl of RISUG as bilateral vas intraluminal injection. After the single-dose administration, the individuals were followed in respect of general health and semen parameters. Their wives were also followed particularly to determine onset of pregnancy. Results: During the six month follow up, the health of male volunteers and their wives was normal with no significant adverse effects. Temporary scrotal enlargement and mild scrotal and inguinal region pain were manifested in most individuals and resolved within one month without any routine activity impairment. In six individuals, there was injection procedure failure and azoospermia was not achieved. The other 133 individuals had either severe oligozoospermia or azoospermia at the first semen examination one month following RISUG injection; 82.7 per cent individuals had continued azoospermia in the month following first semen examination onwards and the rest 17.3 per cent manifested azoospermia within three to six months. Interpretation & conclusions: RISUG intravasal injection appears to be a safe clinical procedure with no significant adverse effects and has high sustained contraceptive efficacy. The localized intervention and continued contraceptive action on single-dose administration were significant features of the RISUG technology.


Assuntos
Anticoncepção/métodos , Anticoncepcionais Masculinos/administração & dosagem , Poliésteres/administração & dosagem , Poliestirenos/administração & dosagem , Ducto Deferente/efeitos dos fármacos , Adulto , Animais , Azoospermia/induzido quimicamente , Azoospermia/diagnóstico , Azoospermia/patologia , Anticoncepcionais Masculinos/efeitos adversos , Feminino , Humanos , Injeções , Masculino , Poliésteres/efeitos adversos , Poliestirenos/efeitos adversos , Gravidez , Sêmen/efeitos dos fármacos , Análise do Sêmen , Espermatozoides/efeitos dos fármacos , Espermatozoides/patologia , Cônjuges , Voluntários
11.
Hum Reprod ; 34(10): 1891-1898, 2019 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-31586185

RESUMO

STUDY QUESTION: Can Chlamydia be found in the testes of infertile men? SUMMARY ANSWER: Chlamydia can be found in 16.7% of fresh testicular biopsies and 45.3% of fixed testicular biopsies taken from a selection of infertile men. WHAT IS KNOWN ALREADY: Male chlamydial infection has been understudied despite male and female infections occurring at similar rates. This is particularly true of asymptomatic infections, which occur in 50% of cases. Chlamydial infection has also been associated with increased sperm DNA damage and reduced male fertility. STUDY DESIGN, SIZE, DURATION: We collected diagnostic (fixed, n = 100) and therapeutic (fresh, n = 18) human testicular biopsies during sperm recovery procedures from moderately to severely infertile men in a cross-sectional approach to sampling. PARTICIPANTS/MATERIALS, SETTING, METHODS: The diagnostic and therapeutic biopsies were tested for Chlamydia-specific DNA and protein, using real-time PCR and immunohistochemical approaches, respectively. Serum samples matched to the fresh biopsies were also assayed for the presence of Chlamydia-specific antibodies using immunoblotting techniques. MAIN RESULTS AND THE ROLE OF CHANCE: Chlamydial major outer membrane protein was detected in fixed biopsies at a rate of 45.3%. This was confirmed by detection of chlamydial DNA and TC0500 protein (replication marker). C. trachomatis DNA was detected in fresh biopsies at a rate of 16.7%, and the sera from each of these three positive patients contained C. trachomatis-specific antibodies. Overall, C. trachomatis-specific antibodies were detected in 72.2% of the serum samples from the patients providing fresh biopsies, although none of the patients were symptomatic nor had they reported a previous sexually transmitted infection diagnosis including Chlamydia. LIMITATIONS, REASONS FOR CAUTION: No reproductively healthy male testicular biopsies were tested for the presence of Chlamydia DNA or proteins or Chlamydia-specific antibodies due to the unavailability of these samples. WIDER IMPLICATIONS FOR THE FINDINGS: Application of Chlamydia-specific PCR and immunohistochemistry in this human male infertility context of testicular biopsies reveals evidence of a high prevalence of previously unrecognised infection, which may potentially have a pathogenic role in spermatogenic failure. STUDY FUNDING/COMPETING INTEREST(S): Funding for this project was provided by the Australian NHMRC under project grant number APP1062198. We also acknowledge assistance from the Monash IVF Group and Queensland Fertility Group in the collection of fresh biopsies, and the Monash Health and co-author McLachlan (declared equity interest) in retrieval and sectioning of fixed biopsies. E.M. declares an equity interest in the study due to financing of fixed biopsy sectioning. All other authors declare no conflicts of interest. TRIAL REGISTRATION NUMBER: N/A.


Assuntos
Azoospermia/microbiologia , Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis/isolamento & purificação , Testículo/microbiologia , Infecções Assintomáticas , Azoospermia/diagnóstico , Azoospermia/patologia , Azoospermia/terapia , Infecções por Chlamydia/complicações , Infecções por Chlamydia/microbiologia , Infecções por Chlamydia/patologia , Chlamydia trachomatis/genética , Estudos Transversais , DNA Bacteriano/isolamento & purificação , Humanos , Masculino , Recuperação Espermática , Testículo/patologia
12.
Fertil Steril ; 112(3): 426-437, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31446902

RESUMO

Over the past few decades, there have been numerous paradigm shifts in male reproductive surgery, resulting from a combination of technologic advancements in both the operating room and the laboratory. The operating microscope transformed the field of male reproductive surgery, enabling novel techniques and higher precision for the treatment of male-factor subfertility. The subsequent widespread adoption of microsurgical approaches was largely responsible for the emergence of a cadre of highly specialized male infertility microsurgeons. The advent and evolution of in vitro fertilization/intracytoplasmic sperm injection was a concurrent story that further revolutionized the field. The ability to achieve fertilization and pregnancy with just a single sperm changed the ways in which male reproductive surgeons could approach a wide range of pathologies from obstructive to nonobstructive causes, culminating in the microdissection testicular sperm extraction procedure for the treatment of nonobstructive azoospermia. Here we review the recent advancements in fertility-enhancing male reproductive surgery for the treatment of four disease processes: varicocele, obstruction of the excurrent ductal system, ejaculatory duct obstruction, and nonobstructive azoospermia. While examining the great strides of the past, we look forward to exciting developments on the horizon.


Assuntos
Preservação da Fertilidade/métodos , Infertilidade Masculina/diagnóstico , Infertilidade Masculina/cirurgia , Azoospermia/diagnóstico , Azoospermia/cirurgia , Preservação da Fertilidade/tendências , Previsões , Humanos , Masculino , Microcirurgia/métodos , Microcirurgia/tendências , Vasovasostomia/métodos , Vasovasostomia/tendências
13.
Hum Reprod ; 34(6): 978-988, 2019 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-31125047

RESUMO

STUDY QUESTION: What is the diagnostic potential of next generation sequencing (NGS) based on a 'mouse azoospermia' gene panel in human non-obstructive azoospermia (NOA)? SUMMARY ANSWER: The diagnostic performance of sequencing a gene panel based on genes associated with mouse azoospermia was relatively successful in idiopathic NOA patients and allowed the discovery of two novel genes involved in NOA due to meiotic arrest. WHAT IS KNOWN ALREADY: NOA is a largely heterogeneous clinical entity, which includes different histological pictures. In a large proportion of NOA, the aetiology remains unknown (idiopathic NOA) and yet, unknown genetic factors are likely to play be involved. The mouse is the most broadly used mammalian model for studying human disease because of its usefulness for genetic manipulation and its genetic and physiological similarities to man. Mouse azoospermia models are available in the Mouse Genome Informatics database (MGI: http://www.informatics.jax.org/). STUDY DESIGN, SIZE, DURATION: The first step was to design of a 'mouse azoospermia' gene panel through the consultation of MGI. The second step was NGS analysis of 175 genes in a group of highly selected NOA patients (n = 33). The third step was characterization of the discovered gene defects in human testis tissue, through meiotic studies using surplus testicular biopsy material from the carriers of the RNF212 and STAG3 pathogenic variants. The final step was RNF212 and STAG3 expression analysis in a collection of testis biopsies. PARTICIPANTS/MATERIALS, SETTING, METHODS: From a total of 1300 infertile patients, 33 idiopathic NOA patients were analysed in this study, including 31 unrelated men and 2 brothers from a consanguineous family. The testis histology of the 31 unrelated NOA patients was as follows: 20 Sertoli cell-only syndrome (SCOS), 11 spermatogenic arrest (6 spermatogonial arrest and 5 spermatocytic arrest). The two brothers were affected by spermatocytic arrest. DNA extracted from blood was used for NGS on Illumina NextSeq500 platform. Generated sequence data was filtered for rare and potentially pathogenic variants. Functional studies in surplus testicular tissue from the carriers included the investigation of meiotic entry, XY body formation and metaphases by performing fluorescent immunohistochemical staining and immunocytochemistry. mRNA expression analysis through RT-qPCR of RNF212 and STAG3 was carried out in a collection of testis biopsies with different histology. MAIN RESULTS AND THE ROLE OF CHANCE: Our approach was relatively successful, leading to the genetic diagnosis of one sporadic NOA patient and two NOA brothers. This relatively high diagnostic performance is likely to be related to the stringent patient selection criteria i.e. all known causes of azoospermia were excluded and to the relatively high number of patients with rare testis histology (spermatocytic arrest). All three mutation carriers presented meiotic arrest, leading to the genetic diagnosis of three out of seven cases with this specific testicular phenotype. For the first time, we report biallelic variants in STAG3, in one sporadic patient, and a homozygous RNF212 variant, in the two brothers, as the genetic cause of NOA. Meiotic studies allowed the detection of the functional consequences of the mutations and provided information on the role of STAG3 and RNF212 in human male meiosis. LIMITATIONS, REASONS FOR CAUTION: All genes, with the exception of 5 out of 175, included in the panel cause azoospermia in mice only in the homozygous or hemizygous state. Consequently, apart from the five known dominant genes, heterozygous variants (except compound heterozygosity) in the remaining genes were not taken into consideration as causes of NOA. We identified the genetic cause in approximately half of the patients with spermatocytic arrest. The low number of analysed patients can be considered as a limitation, but it is a very rare testis phenotype. Due to the low frequency of this specific phenotype among infertile men, our finding may be considered of low clinical impact. However, at an individual level, it does have relevance for prognostic purposes prior testicular sperm extraction. WIDER IMPLICATIONS OF THE FINDINGS: Our study represents an additional step towards elucidating the genetic bases of early spermatogenic failure, since we discovered two new genes involved in human male meiotic arrest. We propose the inclusion of RNF212 and STAG3 in a future male infertility diagnostic gene panel. Based on the associated testis phenotype, the identification of pathogenic mutations in these genes also confers a negative predictive value for testicular sperm retrieval. Our meiotic studies provide novel insights into the role of these proteins in human male meiosis. Mutations in STAG3 were first described as a cause of female infertility and ovarian cancer, and Rnf212 knock out in mice leads to male and female infertility. Hence, our results stimulate further research on shared genetic factors causing infertility in both sexes and indicate that genetic counselling should involve not only male but also female relatives of NOA patients. STUDY FUNDING/COMPETING INTEREST(S): This work was funded by the Spanish Ministry of Health Instituto Carlos III-FIS (grant number: FIS/FEDER-PI14/01250; PI17/01822) awarded to CK and AR-E, and by the European Commission, Reproductive Biology Early Research Training (REPROTRAIN, EU-FP7-PEOPLE-2011-ITN289880), awarded to CK, WB, and AE-M. The authors have no conflict of interest.


Assuntos
Azoospermia/congênito , Proteínas de Ciclo Celular/genética , Testes Genéticos/métodos , Ligases/genética , Meiose/genética , Alelos , Animais , Azoospermia/diagnóstico , Azoospermia/genética , Azoospermia/patologia , Análise Mutacional de DNA/métodos , Bases de Dados Genéticas , Conjuntos de Dados como Assunto , Modelos Animais de Doenças , Estudos de Viabilidade , Sequenciamento de Nucleotídeos em Larga Escala , Homozigoto , Humanos , Masculino , Camundongos , Mutação , Testículo/citologia , Testículo/patologia
15.
Reprod Biomed Online ; 39(1): 134-140, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31097323

RESUMO

RESEARCH QUESTION: Are the outcomes of (i) surgical sperm retrieval (SSR) and (ii) intracytoplasmic sperm injection (ICSI) influenced by the obstructive interval (time elapsed since vasectomy)? DESIGN: Medical records from 148 patients (194 cycles) with secondary azoospermia due to vasectomy, who presented for percutaneous epididymal sperm aspiration (PESA) and ICSI in a private university-affiliated IVF centre, from January 2012 to February 2017, were analysed in this historical cohort study. The obstructive interval was recorded for each couple, and its influences on the outcomes of SSR and ICSI treatment were investigated using general mixed models with adjustment for potential confounders. Clinical pregnancy rate was the main outcome measure. RESULTS: The obstructive interval was negatively correlated with the presence of spermatozoa (ß = -0.032, P = 0.009) and motile spermatozoa (ß = -0.031, P = 0.010) during PESA. The need to convert to testicular sperm aspiration was significantly influenced by the obstructive interval (ß = 0.012, P = 0.003). The blastocyst development rate on day 5 was inversely correlated with the obstructive interval (ß = -0.011, P = 0.014). Implantation and clinical pregnancy rates were negatively influenced by the obstructive interval (ß = -1.107, P = 0.039 and ß = -0.016, P = 0.031, respectively). The receiver operating characteristic curve analysis demonstrated that the obstructive interval has a predictive value on the achievement of clinical pregnancy (area under the curve = 0.667, P = 0.001, Youden index 0.3385, associated criterion >17 years). CONCLUSIONS: Men undertaking vasectomy should be made aware of the long-term effects and their implications for future reproductive treatment.


Assuntos
Infertilidade Masculina/diagnóstico , Infertilidade Masculina/terapia , Taxa de Gravidez , Injeções de Esperma Intracitoplásmicas , Recuperação Espermática , Vasectomia/reabilitação , Adulto , Idoso , Azoospermia/diagnóstico , Azoospermia/etiologia , Azoospermia/terapia , Estudos de Coortes , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Gravidez , Prognóstico , Injeções de Esperma Intracitoplásmicas/estatística & dados numéricos , Recuperação Espermática/estatística & dados numéricos , Fatores de Tempo , Resultado do Tratamento , Vasectomia/efeitos adversos , Adulto Jovem
16.
Fertil Steril ; 112(1): 61-72.e1, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31103287

RESUMO

OBJECTIVE: To study peripheral blood DNA differential methylation in oligozoospermic infertile men in comparison with normozoospermic fertile controls. DESIGN: Case-control study. SETTING: Reproductive biology laboratory. PATIENTS(S): Azoospermic and oligozoospermic infertile patients (n = 6) and normozoospermic fertile controls (n = 6) in the discovery phase, and oligo/asthenozoospermic infertile men (n = 11) and normozoospermic fertile controls (n = 10) in the validation phase. INTERVENTION(S): Blood samples drawn from all participants, DNA isolation and methylation analysis. MAIN OUTCOME MEASURE(S): DNA methylation values analyzed using genomewide methylation 450K BeadChip array, followed by deep sequencing of selected regions for methylation analysis in the neighborhood regions of differentially methylated CpGs. RESULT(S): We found 329 differentially methylated CpG spots, out of which 245 referred to the genes, representing 170 genes. Deep-sequencing analysis confirmed the methylation pattern suggested by 450K array. A thorough literature search suggested that 38 genes play roles in spermatogenesis (PDHA2, PARP12, FHIT, RPTOR, GSTM1, GSTM5, MAGI2, BCAN, DDB2, KDM4C, AGPAT3, CAMTA1, CCR6, CUX1, DNAH17, ELMO1, FNDC3B, GNRHR, HDAC4, IRS2, LIF, SMAD3, SOD3, TALDO1, TRIM27, GAA, PAX8, RNF39, HLA-C, HLA-DRB6), are testis enriched (NFATC1, NMNAT3, PIAS2, SRPK2, WDR36, WWP2), or show methylation differences between infertile cases and controls (PTPRN2, RPH3AL). CONCLUSION(S): We found a statistically significant correlation between peripheral blood DNA methylation and male infertility, raising the hope that epigenome-based blood markers can be used for screening male infertility risk. The study also identified new candidates for spermatogenesis and fertility.


Assuntos
Azoospermia/diagnóstico , Metilação de DNA , Fertilidade/genética , Perfilação da Expressão Gênica/métodos , Análise de Sequência com Séries de Oligonucleotídeos , Oligospermia/diagnóstico , Azoospermia/sangue , Azoospermia/genética , Azoospermia/fisiopatologia , Estudos de Casos e Controles , Ilhas de CpG , Marcadores Genéticos , Predisposição Genética para Doença , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Oligospermia/sangue , Oligospermia/genética , Oligospermia/fisiopatologia , Fenótipo , Valor Preditivo dos Testes
17.
Genet Med ; 21(5): 1209-1217, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31048812

RESUMO

PURPOSE: Comorbid familial nonobstructive azoospermia (NOA) and congenital cataract (CC) have not been reported previously, and no single human gene has been associated with both diseases in humans. Our purpose was to uncover novel human mutations and genes causing familial NOA and CC. METHODS: We performed whole-exome sequencing for two brothers with both NOA and CC from a consanguineous family. Mutation screening of TDRD7 was performed in another similar consanguineous family and 176 patients with azoospermia or CC alone and 520 healthy controls. Histological analysis was performed for the biopsied testicle sample in one patient, and knockout mice were constructed to verify the phenotype of the mutation in TDRD7. RESULTS: Two novel loss-of-function mutations (c.324_325insA (T110Nfs*30) and c.688_689insA (p.Y230X), respectively) of TDRD7 were found in the affected patients from the two unrelated consanguineous families. Histological analysis demonstrated a lack of mature sperm in the male patient's seminiferous tubules. The mutations were not detected in patients with CC or NOA alone. Mice with Tdrd7 gene disrupted at a similar position precisely replicated the human syndrome. CONCLUSION: We identified TDRD7 causing CC as a new pathogenic gene for male azoospermia in human, with an autosomal recessive mode of inheritance.


Assuntos
Azoospermia/genética , Catarata/genética , Ribonucleoproteínas/genética , Adulto , Animais , Azoospermia/diagnóstico , Humanos , Mutação com Perda de Função/genética , Masculino , Camundongos , Camundongos Knockout , Pessoa de Meia-Idade , Mutação , Linhagem , Ribonucleoproteínas/metabolismo , Irmãos , Espermatozoides , Testículo , Sequenciamento Completo do Exoma/métodos
18.
JBRA Assist Reprod ; 23(3): 246-249, 2019 08 22.
Artigo em Inglês | MEDLINE | ID: mdl-30969740

RESUMO

OBJECTIVE: To study the outcomes of testicular sperm extraction (TESE) among men with pure Sertoli cell-only histology identified during diagnostic testicular biopsy. METHODS: This retrospective cohort study involved 1680 cases of patients with nonobstructive azoospermia (NOA) diagnosed with pure Sertoli cell-only histology who underwent testicular biopsy with TESE in a reference center in Brazil by a single surgeon. Sperm retrieval rates (SSR) were the main outcome measure. RESULTS: Overall, 14.83% of patients with Sertoli cell-only had sperm retrieved with TESE in quantity that allowed the performance of ICSI. No differences were observed in SSR based on testis volume (<15 mL vs. <15 mL) or serum FSH level. CONCLUSIONS: Patients with Sertoli cell-only histology can be counseled that they have some likelihood of sperm retrieval with TESE. Based on the findings, patients to be submitted to testicular biopsy for histologic analysis may be concomitantly prepared for ICSI with TESE in case sperm is available.


Assuntos
Síndrome de Células de Sertoli/patologia , Recuperação Espermática , Testículo/patologia , Adulto , Azoospermia/complicações , Azoospermia/diagnóstico , Azoospermia/patologia , Biópsia , Brasil , Técnicas Histológicas , Humanos , Infertilidade Masculina/diagnóstico , Infertilidade Masculina/etiologia , Infertilidade Masculina/patologia , Masculino , Pessoa de Meia-Idade , Tamanho do Órgão , Estudos Retrospectivos , Análise do Sêmen , Síndrome de Células de Sertoli/diagnóstico
19.
Fertil Steril ; 111(5): 873-880, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31029241

RESUMO

Infertility due to obstructive azoospermia may be treated effectively by surgical reconstruction or by retrieval of sperm from the epididymis or testis, followed by in vitro fertilization with intracytoplasmic sperm injection. This replaces the ASRM documents titled "Sperm retrieval for obstructive azoospermia" and "The management of infertility due to obstructive azoospermia," last published in 2008.


Assuntos
Comitês Consultivos , Azoospermia/terapia , Procedimentos Cirúrgicos Reconstrutivos/métodos , Injeções de Esperma Intracitoplásmicas/métodos , Comitês Consultivos/tendências , Azoospermia/diagnóstico , Humanos , Infertilidade Masculina/diagnóstico , Infertilidade Masculina/terapia , Masculino , Microcirurgia/métodos , Microcirurgia/tendências , Procedimentos Cirúrgicos Reconstrutivos/tendências , Injeções de Esperma Intracitoplásmicas/tendências , Recuperação Espermática/tendências
20.
Hum Reprod ; 34(4): 666-671, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30838384

RESUMO

STUDY QUESTION: Are there genetic variants that can be used for the clinical evaluation of azoospermic men? SUMMARY ANSWER: A novel homozygous frame-shift mutation in the MEIOB gene was identified in three azoospermic patients from two different families. WHAT IS KNOWN ALREADY: Up to 1% of all men have complete absence of sperm in the semen, a condition known as azoospermia. There are very few tools for determining the etiology of azoospermia and the likelihood of sperm cells in the testis. The MEIOB gene codes for a single-strand DNA binding protein required for DNA double-strand breaks repair during meiosis. MEIOB appears to be exclusively expressed in human and mouse testis, and MeioB knockout mice are azoospermic due to meiotic arrest. STUDY DESIGN, SIZE, DURATION: Two brothers with non-obstructive azoospermia (NOA) underwent whole-exome sequencing followed by comprehensive bioinformatics analyses. Candidate variations were further screened in infertile and fertile men, as well as in public and local reference databases. PARTICIPANTS/MATERIALS, SETTING, METHODS: This study included 159 infertile and 77 fertile men. The exomes of two Arab men were completely sequenced. In addition, 213 other men of the same Arab ethnicity (136 infertile and 77 fertile men) underwent restriction fragment length polymorphism (RFLP) screening, as did 21 NOA men, of other ethnicities, with testicular impairment of spermatocyte arrest. All of the infertile men underwent Y-chromosome microdeletion and CFTR gene mutation assessments. Comprehensive bioinformatics analyses were designed to uncover candidate mutations associated with azoospermia. MAIN RESULTS AND THE ROLE OF CHANCE: A novel homozygous frame-shift mutation in the MEIOB gene was identified in two brothers of Arab ethnicity. This frame-shift is predicted to result in a truncated MEIOB protein, which lacks the conserved C-terminal DNA binding domain. RFLP screening of the mutation in 157 infertile men, including 112 NOA patients of Arab ethnicity, identified an additional unrelated NOA patient with the same homozygous mutation and a similar testicular impairment. This mutation was not found in available public databases (n > 160 000), nor in the 77 proven fertile men, nor in our database of local Israeli population variations derived from exome and genome sequencing data (n = 500). LIMITATIONS, REASONS FOR CAUTION: We have thus far screened for only two specific MEIOB probable pathogenic mutations in a relatively small local cohort. Therefore, the relative incidence of MEIOB mutations in azoospermia should be further assessed in larger and diverse cohorts in order to determine the efficiency of MEIOB sequence screening for clinical evaluations. WIDER IMPLICATIONS OF THE FINDINGS: The relatively high incidence of likely NOA-causing mutations in MEIOB that was found in our cohort supports the idea that a complete screening of this gene might be beneficial for clinical evaluation of NOA patients. STUDY FUNDING/COMPETING INTEREST(S): This research was supported in part by a grant to EA from the European Research Council under the European Union's Seventh Framework Programme (FP/2007-2013)/ERC grant agreement (616088). There are no competing interests. TRIAL REGISTRATION NUMBER: N/A.


Assuntos
Azoospermia/genética , Proteínas de Ligação a DNA/genética , Meiose/genética , Mutação , Testículo/metabolismo , Adulto , Árabes/genética , Azoospermia/diagnóstico , Azoospermia/etnologia , Azoospermia/patologia , Estudos de Coortes , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Linhagem , Irmãos , Sequenciamento Completo do Exoma
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