Evaluations of cytotoxicity of Smilax myosotiflora and its effects on sexual hormone levels and testicular histology in male rats

Objective: To investigate the cytotoxicity of Smilax myosotiflora (S. myosotiflora) methanolic extract and its effects on sexual hormone levels and testicular histology in male rats. Methods: The cytotoxicity of S. myosotiflora methanolic extract was investigated by employing brine shrimp lethality assay. Forty eight male rats were randomly divided into four groups (Groups I-IV) of 12 each. Rats in Group I were administered with 0.5 mL of distilled water (vehicle), whilst Groups II, III and IV received 200, 400 and 800 mg/kg of the methanolic extract of S. myosotiflora in 0.5 mL of the vehicle, respectively. Male rats treated with continuous daily dosing were killed and necropsied after a total dose period of 60 days. Sexual hormones were assayed and histological examination of testes was performed according to standard methods. Results: S. myosotiflora extracts did not produce any cytotoxicity to brine shrimp in all concentrations tested. Serum testosterone level was significantly higher in rats treated with high dose of S. myosotiflora. Testicular histology showed normal architecture with all stages of spermatogenesis in all experimental groups. Conclusions: The present work confirmed that S. myosotiflora extract improves reproductive functions, without any cytotoxic activity and produces no histological changes to the testes.

MORFOHISTOLOGÍA TESTICULAR DE Hypsiboas pulchellus (AMPHIBIA, HYLIDAE)DURANTE LA ESTACIÓN REPRODUCTIVA

En este estudio se aportan datos sobre la morfohistología testicular de Hypsiboas pulchellus durante el período reproductivo con el fin de conocer aspectos de su biología reproductiva. Los testículos, ovoides y amarillentos de 4,64 ± 0,48 mm de largo por 2,05 ± 0,23 mm de ancho, están rodeados por la túnica albugínea que mide 5,60 ± 0,82 µm. Internamente presentan lóculos seminíferos de 257,47 ± 58,25 µm de diámetro, distinguiéndose en ellos cistos con células espermatogénicas asociadas a células de Sertoli. Existe escaso tejido intersticial. Las espermatogonias I son las células más grandes de la serie germinal (14,34 ± 1,74 µm), estas originan a las espermatogonias II (10,14 ± 1,33 µm). Los espermatocitos I presentan la cromatina levemente condensada midiendo 9,34 ± 0,32 µm. Los espermatocitos II miden 8,12 ± 1,07 µm. Las espermátidas I son esféricas, miden 7,61 ± 1,45 µm, y se agrupan en cistos redondeados. Las espermátidas II, en cambio, son alargadas, no hallándoselas dentro de cistos (4,09 ± 0,51 µm). Los espermatozoides, alargados y flagelados, se hallan libres en el centro del lóculo. La morfohistología de los testículos analizados muestra características similares a las observadas en otros anfibios neotropicales, y presentan todas las células del linaje espermatogénico en un mismo lóculo.

In this study, data concerning to the morphological testicular features of Hypsiboas pulchellus during the breeding season is provided, in order to know aspects of their reproductive biology. The testis, ovoid and yellowish with 4.64 ± 0.48 mm length and 2.05 ± 0.23 mm wide, are surrounded by the tunica albuginea which measures 5.60 ± 0.82 µm. The seminiferous locules, placed internally, measure 257.47 ± 58.25 µm in diameter, and cysts with spermatogenic cells associated with Sertoli cells are distinguished in them. The interstitial tissue is scarce. The biggest cells of the germinal series are the spermatogonia I (14.34 ± 1.74 µm), from which spermatogonia II (10.14 ± 1.33 µm) originate. Spermatocytes I measure 9.34 ± 0.32 µm and have slightly condensed chromatin. The spermatocytes II are 8.12 ± 1.07 µm long. The spermatids I are spherical, grouped in rounded cysts, and measure 7.61 ± 1.45 µm. On the other hand, spermatids II are elongated (4.09 ± 0.51 µm) and not within the cysts. Towards the center of the loculus, the free, lengthened and flagellated spermatozoids are located. The morphohistology of the analyzed testicles show similarities with those observed in other neotropical amphibians, being all the cells of the spermatogenic lineage in the same locule.

Azoospermia y fertilización asistida: rol de la histología testicular en el éxito de icsi / Azoospermia and assisted reproduction: role of testicular histology in the success of ICSI

Introducción: La infertilidad masculina afecta aproximadamente al 7por ciento de los hombres, presentándose hasta el 15 por ciento de ellos con azoospermia. El conocimiento del tipo de azoospermia (obstructiva o no obstructiva) y la localización de la falla (pre-testicular, testicular o post-testicular) es vital para conocer el pronóstico de fertilidad de la pareja y plantear un plan terapéutico adecuado. Actualmente, la extracción de espermatozoides desde epidídimo o testículo de pacientes azoospérmicos, y la posterior inyección intracitoplásmática de éstos (ICSI, por sus siglas en inglés) ha permitido obtener embriones viables para su posterior transferencia. Materiales y métodos: Estudio descriptivo retrospectivo de 42 parejas infértiles con diagnóstico de azoospermia; que se sometieron a biopsia testicular, ICSI y posterior transferencia de embriones, entre los años 2004 y 2012. Se lleva a cabo un análisis de la edad de los pacientes, resultados de la histopatología testicular y su asociación con los resultados de la fertilización asistida. Resultados: 42 pacientes azoospérmicos se sometieron a biopsia testicular y extracción de espermatozoides en el mismo acto quirúrgico. La edad promedio de los pacientes fue de 36 años para los hombres y 32 años para las mujeres. En el análisis histológico de los tejidos testiculares, el 31por ciento de los pacientes presentaban espermatogénesis conservada (EC), el 35.7 por ciento atrofia mixta (AM), el 14.3 por ciento hipoespermatogénesis (HE), el 14.3 por ciento detención de la maduración (DM) y un 4.8 por ciento presentaba otras histologías. Lograron embarazo clínico 14 de 42 parejas (33,3 por ciento), siendo la tasa de embarazo específica por patología de 38,5 por ciento para EC, 26.7 por ciento para AM, 50 por ciento para HE, 16,7 por ciento para DM y 50 por ciento para las otras histologías. 12 de las 42 parejas realizaron más de un ciclo de transferencias...

Introduction: Male infertility affects approximately 7percent of men, presenting up to 15 percent with azoospermia. Knowing the type of azoospermia (obstructive or non-obstructive) and the location of the problem (pre-testicular, testicular and post-testicular) is vital to recognize the fertility prognosis of the couple and present a proper treatment plan. Currently, the extraction of sperm from epididymis or testis of azoospermic patients, and subsequent intracytoplasmic sperm injection (ICSI) has yielded viable embryos for transfer. Materials and Methods: Retrospective study of 42 infertile couples diagnosed with azoospermia, who underwent testicular biopsy, ICSI and subsequent embryo transfer, between 2004 and 2012. We performed an analysis of the patients’ age, testicular histopathology results and their association with assisted fertilization outcome. Results: 42 azoospermic patients underwent testicular biopsy and sperm extraction in the same surgery. The average age of patients was 36 years for men and 32 years for women. Histologic analysis of testicular tissue showed that 31 percent of patients had normal spermatogenesis (NS), 35.7 percent mixed atrophy (MA), 14.3 percent hypospermatogenesis (HS), 14.3 percent maturation arrest (MTA) and 4.8 percent had other histologies. 14 of 42 couples achieved clinical pregnancy (33.3 percent), with a specific pregnancy rate of 38.5 percent for NS, 26.7 percent for MA, 50 percent for HS, 16.7 percent for MTA and 50 percent for other histologies. 12 of 42 couples underwent more than one transfer cycle. Conclusions: The use of ICSI is a suitable alternative for infertile couples with severe male factor. The results of this technique are favorable for most histologies. Patients with MA exhibit sertoli solo syndrome and / or tubular sclerosis foci associated to regions with normal spermatogenesis, enabling the sperm extraction for ICSI.

Investigations on the Factors Predicting the Results of Testis Sperm Extraction / 대한비뇨기과학회지

PURPOSE: Testis sperm extraction and intracytoplasmic sperm injection(TESE-ICSI) has been widely used for the treatment of non-obstructive azoospermia and obstructive azoospermia which cannot be corrected by any other surgical method. We evaluate the results of TESE between groups of obstructive and non-obstructive azoospermia and determine whether there is a significant relationship between the testicular histology, biophysical and endocrine profiles, and the results of TESE in the non-obstructive azoospermia group. MATERIALS AND METHODS: Between 1997 and 1998, a total of 64 patients underwent TESE procedures, including 23 patients with non-obstructive azoospermia and 41 with obstructive azoospermia, who were beyond the reach of surgical correction and microsurgical epididymal sperm aspiration(MESA). We compared TESE results between two groups. We investigated the factors predicting the results of TESE and tried to correlate the results with biophysical, endocrine profiles and testicular histology. RESULTS: Non-obstructive azoospermia group had significantly smaller testes, fewer motile sperm, and lower sperm retrieval rate than obstructive group. Statistically significant differences were detected in plasma FSH, LH, and testis volume according to different histology patterns in the non-obstructive azoospermia group. There were no significant differences in age, testis volume, semen volume, plasma FSH, LH, and testis histology between the patients with successful and failed sperm retrieval in the non-obstructive azoospermia group. CONCLUSIONS: Obstructive azoospermia group is expected to have better chance of pregnancy than non-obstructive azoospermia group due to a difference of sperm retrieval rate. In non-obstructive azoospermia group, sperm retrieval has poor correlations with biophysical and endocrine profiles, and with histological patterns. No single variable can provide a perfect discrimination between the groups with successful and failed TESE. TESE should be performed in carefully selected cases until a reliable criteria predicting the TESE result is established.