Testicular steroid sulfatase overexpression is associated with Leydig cell dysfunction in primary spermatogenic failure.

BACKGROUND: Decreased testosterone (T) to LH ratio and increased 17ß-estradiol (E2) serum concentrations represent a common finding among patients with severe spermatogenic failure, suggesting a concurrent Leydig cell steroidogenic dysfunction. Aromatase overexpression has been associated with increased serum and intratesticular E2 in these patients. However, it is unknown whether the sulfatase pathway contributes to the increased availability of active estrogens in patients with primary spermatogenic failure. OBJECTIVES: To assess estrogen sulfotransferase (SULT1E1) and steroid sulfatase (STS) mRNA abundance in testicular tissue of patients with Sertoli cell-only syndrome (SCOS) and normal tissues, its association with serum and intratesticular hormone levels, and to explore the mRNA and protein testicular localization of both enzymes. MATERIALS AND METHODS: Testicular tissues of 23 subjects with SCOS (cases) and 22 patients with obstructive azoospermia and normal spermatogenesis (controls) were obtained after biopsy. SULT1E1 and STS transcripts accumulation was quantified by RT-qPCR. For mRNA and protein localization, we performed RT-qPCR in Leydig cell clusters and seminiferous tubules isolated by laser-capture microdissection and immunofluorescence in testicular tissues. Serum and intratesticular hormones were measured by immunoradiometric assays. RESULTS: SULT1E1 mRNA accumulation was similar in both groups. The amount of STS mRNA was higher in cases (p = 0.007) and inversely correlated with T/LH ratio (r = -0.402; p = 0.02). Also, a near significant correlation was observed with intratesticular E2 (r = 0.329, p = 0.057), in agreement with higher intratesticular E2 in cases (p < 0.001). Strong STS immunoreaction was localized in the wall of small blood vessels but not in Leydig cells. Both SULT1E1 and STS mRNA abundance was similar in Leydig cell clusters and the tubular compartment, except for lower SUTL1E1 mRNA in the seminiferous tubules of SCOS patients (p = 0.001). CONCLUSIONS: Our results suggest that an unbalance of the STS/SULT1E1 pathway contributes to the testicular hyperestrogenic microenvironment in patients with primary spermatogenic failure and Leydig cell dysfunction.

Partial-AZFc deletions in Chilean men with primary spermatogenic impairment: gene dosage and Y-chromosome haplogroups.

PURPOSE: To investigate the association of partial-AZFc deletions in Chilean men with primary spermatogenic failure and their testicular histopathological phenotypes, analyzing the contribution of DAZ dosage, CDY1 copies, and Y-chromosome haplogroups. SUBJECTS AND METHODS: We studied 479 Chilean men: 334 infertile patients with histological examination (233 cases with spermatogenic defects and 101 normal spermatogenesis, obstructive controls, OC), and 145 normozoospermic controls (NC). AZFc subdeletions were detected by single-tagged sequences and single nucleotide variants analysis. DAZ-copy number was quantified by real-time qPCR. Y-chromosome haplogroups (Y-hg) were hierarchically genotyped through 16 biallelic-markers. RESULTS: The prevalence of AZFc-partial deletions was increased in cases (6%) compared with NC (1.4%) (P = 0.035). There was no difference between 143 Sertoli-cell only syndrome, 35 maturation arrest, or 35 mix atrophy patients and controls. However, gr/gr deletions were more frequent in 16 subjects with hypospermatogenesis compared with NC (P = 0.003) and OC (P = 0.013). Y-hg R was the most prevalent (~ 50%), but decreased among gr/gr deletions (21%, P = 0.03). The prevalence of Y-hg M increased in cases versus controls, both in total and non-deleted men (3.9 and 3.7% versus 0.4%, P = 0.009 and P = 0.016, respectively). Among gr/gr deletions, Y-hg H increased compared with non-deleted men (14.3% versus 0.4%, P = 0.0047). CONCLUSION: Partial-AZFc deletions in a Chilean admixed population are associated with secretory azo/oligozoospermia and might have a role in the development of hypospermatogenesis. Low represented haplogroups, Y-hg M and Y-hg H, show an association with the occurrence of spermatogenic failure and gr/gr deletions respectively; however, additional studies are required.

Caracterización de población masculina que consulta por infertilidad: experiencia local de 10 años / Caracterization of menpopulation with infertility: 10 years local experience

Introducción: La Infertilidad ha mostrado un aumento progresivo de su prevalencia. Se estima que alcanza un 15 porciento de las parejas, con un 35 porciento a 40 porciento de componente masculino y un 20 a 30 porciento afectando a ambos miembros. El objetivo de este estudio es la descripción de las características de los hombres que consultan por infertilidad a nivel local así como los resultados de sus estudios iniciales.Material y métodos. Estudio transversal descriptivo en donde se revisaron fichas de 500 pacientes de sexo masculino que consultaron por infertilidad entre enero de 2006 y junio de 2016. De cada paciente se registró edad, paridad previa, frecuencia sexual, factores de riesgo como antecedentes de criptorquidea, varicocele, cáncer, entre otros. Además se registraron los resultados de estudios realizados. Los datos fueron analizados con el sistema estadístico STATA 13.0.Resultados. La mediana de edad fue de 34 años (15-68 años) con un 70,29 porciento de infertilidad primaria. Respecto al tiempo de exposición la mediana fue de 7 meses (3-31) y el promedio de frecuencia sexual fue de 2,5 veces por semana (0-7). En un 48,69 porciento se encontró un examen genital alterado, un 9,32 porciento tuvo un desarrollo puberal alterado, y el antecedente de criptorquidea y varicocele estuvo presente en un 15 porciento y 23,2 porciento respectivamente. El 18,4 porciento presentó alguna disfunción sexual, el 46 porciento era tabáquico y un 8,77 porciento consumía marihuana. Respecto al estudio se observaron 2 casos de microdeleciones del cromosoma Y, y 2 casos de cariogramas alterados. Se rescataron 254 espermiogramas con un 67,32 porciento de azospermicos, de los cuales un 14,62 porciento fueron obstructivos.Conclusión. El componente masculino en la infertilidad requiere de una evaluación anamnésica prolija debido a la variedad de factores de riesgo involucrados, siendo importante la pesquisa de condiciones que puedan intervenirse precozmente para así lograr mayores tasas de éxito en la recuperación de espermatozoides y posterior técnica de fertilidad. Existen escasos datos epidemiológicos de infertilidad en Chile, por lo que este trabajo nos permite conocer el perfil de pacientes que consultan por esta patología a nivel local.(AU)

Introduction: Infertility prevalence has shown a progressive increase. It is estimated that it reaches 15 pertcent of the couples, with 35 pertcent to 40 pertcent of male component and 20 to 30 pertcent affecting both members. The objective of this study is to describe the characteristics of men who consult for infertility in our center.Material y Methods. A descriptive cross-sectional study was carried out in which we reviewed the records of 500 male patients who consulted for infertility between January 2006 and June 2016. It registered of each patient, age, previous parity, sexual frequency, risk factors such as history of cryptorchid, varicocele, Cancer, among others. In addition, the results of studies performed were recorded. The data were analyzed using the statistical system STATA 13.0.Results. The median age was 34 years (15-68 years) with 70.29 pertcent of primary infertility. Regarding the exposure time the median was 7 months (3-31) and the average sexual frequency was 2.5 times per week (0-7). In 48.69 pertcent an altered genital examination was found, 9.32 pertcent had an altered pubertal development, and the antecedent of cryptoquidea and varicocele was present in 15 pertcent and 23.2 pertcent respectively. 18.4 pertcent had sexual dysfunction, 46 pertcent were smokers and 8.77 pertcent used marijuana. Regarding the exams, 2 cases of microdeletions of the Y chromosome were observed, and 2 cases of Klinefelter Syndrome. 254 spermiograms were rescued with 67.32 pertcent of azoospermia, of which 14.62 pertcent were obstructive.Conclusion. The male component in infertility requires a complete anamnestic evaluation due to the variety of risk factors involved, being important the research of conditions that can intervene early to achieve higher rates of success in sperm retrieval and subsequent fertility technique. There are few epidemiological data on infertility in Chile, so this work allows us to know the profile of patients who consult for this pathology at the local population.(AU)

Levels of the retinoic acid synthesizing enzyme aldehyde dehydrogenase-1A2 are lower in testicular tissue from men with infertility.

OBJECTIVE: To determine whether decreased testicular levels of enzymes necessary for retinoic acid biosynthesis were associated with male infertility, as retinoic acid is known to be necessary for spermatogenesis. DESIGN: Observational analysis of testicular tissue samples, sperm indices, and serum hormone concentrations. SETTING: Two infertility centers in Chile. PATIENT(S): 32 infertile men and 11 control men. INTERVENTION(S): Measurement of the three enzymes necessary for retinoic acid biosynthesis, aldehyde dehydrogenase (ALDH) 1A1, 1A2, and 1A3, in testicular tissue by a novel liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) peptide assay. MAIN OUTCOME MEASURE(S): ALDH isozyme levels compared by type of infertility and correlated with testicular germ cell numbers, sperm parameters, and serum and intratesticular hormone concentrations. RESULT(S): Men with infertility had statistically significantly reduced levels of ALDH1A2 but not ALDH1A1 or ALDH1A3 in their testicular tissue compared with men with normal spermatogenesis. The ALDH1A2 protein levels were strongly correlated with the number of germ cells found via testicular biopsy. CONCLUSION(S): These findings suggest that ALDH1A2 is the enzyme involved in retinoic acid biosynthesis in human germ cells. Further study of the relationship between intratesticular ALDH1A2 and male infertility is warranted to determine whether men with infertility have a reduced ability to synthesize retinoic acid within their germ cells that could impair spermatogenesis.

Greater prevalence of Y chromosome Q1a3a haplogroup in Y-microdeleted Chilean men: a case-control study.

PURPOSE: To determine the prevalence of South Amerindian Y chromosome in Chilean patients with spermatogenic failure and their association with classical and/or AZFc-partial Y chromosome deletions. METHODS: We studied 400 men, 218 with secretory azo/oligozoospermia (cases) and 182 controls (116 fertile and/or normozoospermic, and 66 azoospermic with normal spermatogenesis). After a complete testicular characterization (physical evaluation, hormonal and/or biopsy) peripheral blood was drawn to obtain DNA for Y chromosome microdeletions, AZFc-partial deletions and biallelic analysis by allele specific polymerase chain reaction (PCR) of the M3 (rs3894) single nucleotide polymorphism (SNP). RESULTS: Classical AZF microdeletions were found in 23 cases (Y-microdeleted). AZFc-partial deletions were observed in 10 cases (6 "gr/gr", 3 "b2/b3" and 1 "b1/b3") and 4 controls (4 "gr/gr"). The AZFc-partial deletions were mainly associated with the absence of DAZ1/DAZ2 (64 %). No significant differences in the prevalence of AZFc-partial deletions were observed between cases and controls. We observed a significant higher proportion of the Q1a3a haplogroup in Y-microdeleted men compared to patients with spermatogenic failure without deletions and control men (P<0.01 and P<0.05, respectively by Bonferroni test). Among them, patients with AZFb deletions had an increased prevalence of the Q1a3a haplogroup compared to controls, cases without deletions and to those with complete or partial-AZFc deletions (P<0.01, Bonferroni test). CONCLUSIONS: The Q1a3a South Amerindian lineage seems to increase the susceptibility to non AZFc microdeletions. On the other hand, in Chilean population the AZFc-partial deletions ("gr/gr", "b1/b3" and/or "b2/b3") does not seem to predispose to severe spermatogenic impairment.

Analysis of 6 single-nucleotide polymorphisms in the androgen receptor gene in Chilean patients with primary spermatogenic failure.

Androgens are essential for spermatogenesis. It has been postulated that androgen activity is modulated directly or indirectly by genetic variability in the androgen receptor gene sequence, including CAG/GGN polymorphisms and single-nucleotide polymorphisms (SNP). In this study, the frequency of 6 SNPs that constitute a haplotype in the androgen receptor sequence was determined by enzyme restriction assays and allele-specific polymerase chain reactions in 117 secretory azo/oligozoospermic men (93 idiopathic and 24 excryptorchidic), and in 121 controls with normal spermatogenesis (42 obstructive and 79 normozoospermic men) whose hormonal measurements and length of CAG/GGN polymorphisms were previously determined. The frequency of these 6 SNPs was not different between patients and controls. A total of 10 haplotypes (HAPs 1-10) formed by these 6 SNPs were found, and one of these haplotypes was observed with high frequency in the total population (HAP1, 83.2%; P < .001, χ(2) test). The frequency of the 10 haplotypes was not different between patients and controls, except for HAP5, which was only detected in one patient with a history of bilateral cryptorchidism (P = 0.014, Bonferroni test). On the other hand, no associations were found between the haplotypes studied and shorter or longer CAG or GGN polymorphisms. Interestingly, we found that the CAG 21 allele, which was previously correlated with an increased risk of idiopathic spermatogenic impairment, was more frequently found among the less common haplotypes that have higher follicle-stimulating hormone serum levels. In summary, we did not find an increased frequency of particular haplotypes in infertile men with idiopathic spermatogenic impairment compared with control men; however, we found that the CAG 21 allele, which appears to be associated with male infertility, is observed at a significantly higher proportion among the less common androgen receptor haplotypes.

Linfoma testicular: incidencia, descripción epidemiológica y manejo durante el período 2005-2011 / Testicular lymphoma: incidence, epidemiologic description and management during the 2005-2011 period

El linfoma testicular es una patología infrecuente, correspondiendo al 9 por ciento de los cánceres testiculares, presentándose más frecuentemente entre los 60 a 80 años (25-50 por ciento). La presentación clínica más frecuente es el aumento de volumen unilateral e indo/oro. El tipo histológico más común es linfoma difuso de células grandes B (60-90 por ciento). La orquidectomía radical asociada a quimioterapia y radioterapia es la primera línea de tratamiento para los pacientes con enfermedad limitada. Material y método: Estudio retrospectivo descriptivo. Se revisó y filtró la lista de pacientes ingresados al SIGGES como tumor testicular entre enero 2005 a abril 2011. De los pacientes con diagnóstico histológico e inmunohistoquímico compatible, se registraron las características epidemiológicas, estudio, manejo y sobrevida. Posteriormente se realizó un análisis de la base de datos con el programa estadístico SPSS 13. 0. Resultados: De un total de 299 pacientes con el diagnóstico histológico de cáncer testicular, 8 pacientes fueron diagnosticados como linfoma testicular confirmado por histología e inmunohistoquímica. El promedio y mediana de edad fue 52 años y 63 años (18-73) respectivamente. Tres casos (37,5 por ciento) correspondieron a presentaciones secundarias. En 6 de los casos (75 por ciento) el testículo afectado fue el derecho. Histológicamente, el 63 por ciento correspondió a Linfoma difuso de células grande B. Clínicamente, el todos los casos se presentaron con aumento de volumen y con marcadores en rango normal. En 7 casos (8 7, 5 por ciento) el diagnóstico y manejo inicial fue mediante orquidectomía radical, y en un caso por biopsia testicular, con orquidectomía posterior 3 casos presentaron diseminación...

esticular lymphoma is a rare disease, happening in 9 percent of testicular cancers, most commonly between the ages 60 to 80 years (25 percent-50 percent). The most common presentation is unilateral indolent testicular growth. Histology shows a diffuse big B cell lymphoma in most of the cases (60 percent-90 percent). Radical orchiectomy, chemotherapy and radiation are the first line therapy for patients with limited disease. Materials and methods: Retrospective clinical study. We included and filtered the SIGGES list of patients admitted for Testicular Tumor from January 2005 to April 2011. Patients with a compatible diagnosis were analyzed, using SPSS 13.0® as statistical software. Result: Of a total number of 299 testicular cancer patients 8 presented with a histological and inmunnohistochemical testicular lymphoma. Mean age was 52 years and the median 63 years (18-73). ln three cases (37.5 percent) it was a secondary localization. ln 6 cases ( 75 percent) the affected testicle was the right one. 63 percent corresponded to a diffuse big cell B cell Lymphoma. All patients presented normal tumor markers. ln 7 (87,5 percent) cases the initial treatment was radical orchiectomy in one patient the diagnosis was don through a testicular biopsy, and the orchidectomy was differed. 3 cases presented dissemination. In 7 patients adjuvant chemotherapy was performed. Mortal/ty was 38 percent with a 1 7-month follow-up. Conclusion: Testicular lymphoma is a rare condition with bad prognosis. Histology is fundamental for treatment, an in this sense inmunohystochemcal analysis is especially helpful...

Cirugía percutánea renal bilateral simultánea en decúbito supino: experiencia inicial / Percutaneous bilateral renal surgery in supine position: initial experience

La litiasis renal bilateral voluminosa es un desafío quirúrgico para el endourólogo. La cirugía percutánea bilateral simultánea (CPRBS) es una modalidad aceptada, tanto en adultos como niños, sin embargo existe poca literatura al respecto. Presentamos nuestra experiencia de 3 pacientes (6 unidades renales) en posición de Valdivia-Galdakao. Incluimos el primer reporte mundial de un caso de CPRBS tubeless bilateral en de cúbito supino...

Simultaneous bilateral PCNL (SBPCNL) is a safe procedure and can be used effectively in adults as well as in children. In addition to being cost effective, it involves only a single anesthesia with a shorter hospital stay and faster convalescence. We present our initial experience of 3 cases (6 renal units) in supine position. We include the first report to our knowledge of a tubeless SBPCNL in supine position...

46,XX/SRY-negative true hermaphrodite.

OBJECTIVE: To describe genetic evaluation and response to surgery and letrozole therapy of a 46,XX/SRY-negative true hermaphrodite. DESIGN: Case report. SETTING: University Medical Center. PATIENT(S): Nineteen-year-old male with penile hypospadias, micropenis, and crytorchidism at the time of birth. INTERVENTION(S): Unilateral gonadectomy, and contralateral conservative gonadal surgery, followed by therapy with letrozole. MAIN OUTCOME MEASURE(S): Histopathologic, genetic and hormonal studies. RESULT(S): Genetic analysis showed that the subject was 46,XX/SRY-negative. Gonadectomy of the left gonad was performed at 16 years. The gonad resected was an ovotestes. The patient's estradiol was high (492±25 pmol/L), whereas the testosterone was low (4.2±0.5 nmol/L). Nineteen months later, conservative gonadal surgery of the contralateral gonad was performed to resect ovarian tissue, and treatment with letrozole was started. During letrozole treatment, testosterone was significantly increased (8±0.7 nmol/L), but estradiol was not changed (323±118 pmol/L). After letrozole withdrawal, testosterone did not decreased significantly (6.9±0.4 nmol/L), estradiol showed an oscillating pattern and a gonadal ultrasound showed an ovoid structure, which appeared to correspond to a follicle. At that time, estradiol was elevated (393 pmol/L). CONCLUSION(S): We present the case of a 46,XX/SRY-negative phenotypic male with bilateral ovotestes. Conservative gonadal surgery should be performed only when all ovarian tissue can be resected. Our results suggest that letrozole is not an adequate treatment for 46,XX true hermaphrodite males with ovotestes.

Androgen receptor gene CAG and GGN repeat polymorphisms in Chilean men with primary severe spermatogenic failure.

There is ample documentation supporting the fact that androgens are required for normal spermatogenesis. A minority of infertile men have abnormal testosterone blood levels or mild androgen receptor mutations. We investigated the androgen receptor CAG and GGN repeat lengths in Chilean men with spermatogenic impairment. We studied 117 secretory azoospermic/oligozoospermic men (93 idiopathic and 24 excryptorchidic), without Y-chromosome microdeletions, and 121 controls with normal spermatogenesis (42 obstructive and 79 normozoospermic men). Peripheral blood was drawn to obtain genomic DNA for polymerase chain reaction and automated sequencing of CAG and GGN repeats. Testicular characterization included hormonal studies, physical evaluation, and seminal and biopsy analysis. The CAG and GGN polymorphism distributions were similar among idiopathic men, excryptorchidic men, and controls and among the different types of spermatogenic impairment. However, the proportion of the CAG 21 allele was significantly increased in idiopathic cases compared to controls (P = .012 by Bonferroni test, odds ratio = 2.99, 95% confidence interval, 1.27-7.0) and the CAG 32 allele only was observed in excryptorchidic patients (P < .0002, Bonferroni test). Idiopathic cases with Sertoli cell-only syndrome showed the highest proportion of the CAG 21 allele (P = .024, χ(2) test). On the other hand, in idiopathic cases and controls the most common GGN allele was 23, followed by 24, but an inverse relation was found among excryptorchidic cases. The joint distribution of CAG and GGN in control, idiopathic, and excryptorchidic groups did not show an association between the 2 allele repeat polymorphisms (P > 0.05, χ(2) test). Our results suggest that the CAG 21 allele seems to increase the risk of idiopathic Sertoli cell-only syndrome. Moreover, the GGN 24 allele could be contributing to deranged androgen receptor function, associated with cryptorchidism and spermatogenic failure.

AZFc partial deletions in Chilean men with severe spermatogenic failure.

OBJECTIVE: To determine the prevalence of AZFc subdeletions in infertile Chilean men with severe spermatogenic impairment. DESIGN: Prospective analysis. SETTING: University infertility clinic. PATIENT(S): Ninety-five secretory azo/oligozoospermic men without AZFc Y chromosome microdeletions: 71 whose testicular histology showed severe spermatogenic impairment and 24 who exhibited reduced testicular volume and elevated serum FSH levels. As controls, we studied 77 men (50 fertile and/or normozoospermic, and 27 with azoospermia and normal spermatogenesis). INTERVENTION(S): Peripheral blood was drawn to obtain genomic DNA for polymerase chain reaction (PCR) digestion assays of DAZ-sequence nucleotide variants and for AZFc-STS PCR after a complete testicular characterization (biopsy, hormonal, and physical evaluation). MAIN OUTCOME MEASURE(S): DAZ genes and AZFc subdeletion types. RESULT(S): In cases we observed two "gr/gr" subdeletions (2.1%), one with absence of DAZ1/DAZ2 (g1/g2 subtype), and the other with absence of DAZ3/DAZ4 (r2/r4 subtype). Additionally, we found a g1/g3 subdeletion in a patient with Sertoli-cell-only syndrome. In controls, we observed two gr/gr subdeletions with absence of DAZ1/DAZ2 (2.6%) in a fertile/normozoospermic and in an obstructive azoospermic man. CONCLUSION(S): AZFc subdeletions do not seem to cause severe impairment of spermatogenesis. Moreover, gr/gr-DAZ1/DAZ2 subdeletions do not appear to affect fertility in Chilean men.