Cisplatin-induced azoospermia and testicular damage ameliorated by adipose-derived mesenchymal stem cells

BACKGROUND: The testes are highly susceptible to the adverse effects of chemotherapy and radiation at all stages of life. Exposure to these threats mainly occurs during cancer treatment and as an occupational hazard in radiation centers. The present study investigated the regenerative ability of adipose-derived mesenchymal stem cells (ADMSCs) against the adverse effects of cisplatin on the structure and function of the testes. METHODS: New Zealand white rabbits (N = 15) were divided into three groups of five: a negative control group (no treatment), a cisplatin group (single dose of cisplatin into each testis followed three days later by a PBS injection), and a cisplatin + ADMSCs group (cisplatin injection followed three days later by an ADMSC injection). On day 45 post-treatment, serum testosterone levels were evaluated, and the testes and epididymis were collected for histology, oxidative stress examination, and epididymal sperm analysis. RESULTS: Cisplatin caused damage to the testicular tissue and decreased serum testosterone levels, epididymal sperm counts, and oxidants. An antioxidant imbalance was detected due to increasing malondialdehyde (MDA) and reduced glutathione (GSH) levels in testicular tissue. The ADMSC-treated group displayed a moderate epididymal sperm count, adequate antioxidant protection, suitable hormone levels, and enhanced testicular tissue morphology. CONCLUSIONS: ADMSCs treatment repaired damaged testicular tissue, enhanced biochemical parameters, and modified pathological changes caused by cisplatin.

Genetic pathogenesis of acephalic spermatozoa syndrome: past, present, and future / 亚洲男科学杂志(英文版)

Acephalic spermatozoa syndrome (ASS) is one of the most severe spermatogenic failures of all infertility in men. The cognition of ASS has experienced a tortuous process. Over the past years, with the in-depth understanding of spermatogenesis and the emergence of new genetic research technologies, the unraveling of the genetic causes of spermatogenic failure has become highly active. From these advances, we established a genetic background and made significant progress in the discovery of the genetic causes of ASS. It is important to identify pathogenic genes and mutations in ASS to determine the biological reasons for the occurrence of the disease as well as provide genetic diagnosis and treatment strategies for patients with this syndrome. In this review, we enumerate various technological developments, which have made a positive contribution to the discovery of candidate genes for ASS from the past to the present. Simultaneously, we summarize the known genetic etiology of this phenotype and the clinical outcomes of treatments in the present. Furthermore, we propose perspectives for further study and application of genetic diagnosis and assisted reproductive treatment in the future.

Research progress on the correlation between abnormal sperm DNA methylation and semen quality / 中华医学遗传学杂志

DNA methylation as an important aspect of epigenetics plays an important role in spermatogenesis and embryonic development. In recent years, researchers have found that male infertility, in particular abnormal semen quality, is related to abnormal DNA methylation. To further delineate the pathogenesis of male infertility and inspire new ideas for the treatment of male infertility, a comprehensive review over the correlation between abnormal methylation of imprinted genes, repetitive DNA elements and non-imprinted genes, semen quality (including sperm count, morphology, and vitality) and male infertility is provided.

Are specialized sperm function tests clinically useful in planning assisted reproductive technology?

CASE STUDY 40-year-old male patient and 32-year-old female partner, with a history of primary infertility of two years duration. The workup revealed idiopathic mild oligoasthenotheratozoospermia, and no apparent female infertility factors. The couple has failed three intrauterine insemination (IUI) cycles, planning more IUI cycles but also considering in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI).

High exposure to boron in drinking water and sperm parameters in Chilean young people / Exposición elevada a boro en el agua potable y parámetros de esperma en gente joven chilena

Our aim was to describe sperm parameters in residents from Northern Chile. We evaluated in 101 volunteers (18 and 30 years old) urinary and drinking water Boron levels using Inductively Coupled Plasma Mass Spectrometry; semen parameters were measured with standardized methods. Each individual was categorized in 3 levels of exposure: low (B levels in urine 2.94 mgL-1 or tap water 3.0 mgL-1), medium (urinary B between 2.95-7.4 mgL-1 and B in tap water with 3.0-7.0 mgL-1) and high (urinary B > 7.4 mgL-1 or tap water > 7.0 mgL-1). We found no significant differences among groups by pH, sperm concentration (45.1; 48.2 and 38 million/mL), motility 1th hour (38.1; 40.0 and 45.5 %) and vitality 1th hour (88.6; 88.0 and 76.9 %) respectively. Abnormal morphology was significant different (83.3; 90 and 83 %). Young men exposed to B in drinking water present sperm variations associated with the level of exposure. Most of these changes are positive at intermediate levels of B. For the highest exposures were observed negative changes in sperm morphology, concentration, motility and vitality, all relevant parameters of fertility. Beneficial effect is observed at medium exposure, like a "U curve".

El objetivo de este trabajo fue describir los parámetros espermáticos en residentes del norte de Chile. Se evaluaron en 101 voluntarios (18 y 30 años), los niveles urinarios y de agua potable de boro, usando "Inductively Coupled Plasma Mass Spectrometry". Los parámetros del semen se midieron con métodos estandarizados. Cada individuo se clasificó en 3 niveles de exposición: bajo (niveles B en la orina 2,94 mgL-1 o agua potable 3,0 mgL-1), medio (B urinario entre 2,95-7,4 mgL-1 y B en agua de beber con 3,0- 7,0 mgL-1) y alto (B urinario >7,4 mgL-1 o agua potable > 7,0 mgL-1). No se encontraron diferencias significativas entre los grupos por pH, concentración de espermatozoides (45,1; 48,2 y 38 millones/mL), motilidad a 1 hora (38,1; 40,0 y 45,5%) y vitalidad 1 hora (88,6; 88,0 y 76,9%) respectivamente. La morfología anormal fue significativamente diferente (83,3; 90 y 83%). Los hombres jóvenes expuestos a B en el agua potable presentan variaciones espermáticas asociadas con el nivel de exposición. La mayoría de estos cambios son positivos en niveles intermedios de B. Para las exposiciones más altas se observaron cambios negativos en la morfología, concentración, motilidad y vitalidad del esperma, parámetros relevantes de la fertilidad. Un efecto beneficioso se observa en la exposición media, como una "curva U".

Epididymal morphophysiology of adult collared peccaries (Pecari tajacu) / Morfofisiologia epididimária de catitus (Pecari tajacu) adultos

A functional understanding of the epididymis makes it possible to increase a species' fertility, since it facilitates the preservation of gametes for use in assisted reproduction techniques. This study analyzed the histological features of the different regions of the epididymis of the Pecari tajacu and the associated pathologies found in the spermatozoa present in this organ. After an orchiectomy surgery to remove the testicles and epididymis obtained from eight adult animals bred in captivity, fixation and histological processing of samples were carried out. The presence of pseudostratified columnar epithelium with stereocilia was observed in all regions of the epididymis (caput, corpus and caudal). Similarities were noted between the average height of the corpus and caudal epithelium, which differed from that found at the caput (1- proximal: region associated with the efferent ducts; 2- distal: region associated with corpus epididymis). It was also observed that the proximal caput presents a smaller average of the tubules diameter compared to other portions of the epididymis. Regarding pathologies of the spermatozoa, thirteen different types of morphological modifications were observed. Underdeveloped spermatozoa and detached heads were most frequently encountered along the epididymis of the collared peccaries. This study is a pioneer in the area and its data will serve as a basis for comparative research on the reproductive biology of artiodactyls, thus contributing to their conservation and reproduction.(AU)

O entendimento funcional do epidídimo possibilita o aumento da fertilidade de uma espécie, pois torna possível a preservação de gametas para o uso em técnicas de reprodução assistida. Este trabalho estudou os aspectos histológicos das diferentes regiões do epidídimo de Pecari tajacu, bem como as patologias associadas encontradas nos espermatozoides presentes nesse órgão. Após procedimento cirúrgico de orquiectomia, para remoção dos testículos e epidídimos, obtidos de oito animais adultos nascidos em cativeiro, realizou-se a fixação e o processamento histológico das amostras. Constatou-se em todas as regiões do epidídimo (cabeça, corpo e cauda) a presença de epitélio pseudoestratificado colunar com estereocílios. Foram observadas semelhanças entre a altura média do epitélio do corpo e da cauda, a qual era diferente da encontrada na cabeça (1- proximal: região associada aos ductos eferentes; 2- distal: região associada ao corpo do epidídimo). Foi verificado que a cabeça proximal apresenta a média de diâmetro dos túbulos menor em relação às demais porções do epidídimo. Em relação às patologias dos espermatozoides, constataram-se treze tipos diferentes de alterações morfológicas. Os espermatozoides subdesenvolvidos e cabeças soltas foram as mais frequentes ao longo do epidídimo de catitus. Os dados deste trabalho são pioneiros, e servirão como base para pesquisas comparativas sobre a biologia reprodutiva dos artiodátilos, contribuindo para a sua conservação e produção.(AU)

Melatonin protective role in mouse cauda epipidymal spermatozoa damage induced by sodium arsenite / Rol protector de la melatonina en el daño de espermatozoides de cauda epididimaria en ratón producido por arsenito de sodio

We evaluated the sperm parameters such as cauda epididymis weight, sperm count, sperm morphology and sperm DNA stability of adult CF-1 male mice treated daily (oral exposure) with the toxic sodium arsenite (As, 7.0 mg/kg/body weight); Melatonin (Me, 10.0 mg/kg/bw), Me (10.0 mg/kg/bw) plus As (7.0 mg/kg/bw) and Negative Control (NaCl 0.9 percent) to assess acute (8.3 days), chronic (33.2 days) and recovery of testicular damage (66.4 days). Arsenic decreases the number of sperm from chronic treatment (33.2 days) and this effect continued until 66.4 days of treatment. The toxic effect of As also altered the morphology of spermatozoa in all treatment periods when compared to the negative control group. However, Metalonin induced protective effects in periods of 33.2 and 66.4 days of treatment. Additionally, the stability of DNA was significantly affected by arsenic in all periods, but the chronic treatment (33.2 days) in the AsMe revealed increased stability compared to the group treated with arsenic only. Melatonin partially protects sperm toxicity caused by Arsenic, especially during periods of 33.2 and 66.4 days.

Se evaluaron los parámetros espermáticos como peso de la cola del epidídimo, conteo de espermatozoides, morfología de los espermatozoides y estabilidad del ADN de espermatozoides de ratones machos adultos CF-1 tratados diariamente (exposición oral) con el tóxico arsenito de sodio (As, 7,0 mg/kg/peso corporal), melatonina (Me, 10,0 mg/kg/pc, Me (10,0 mg/kg/pc) más As (7,0 mg/kg/pc) y el Control Negativo (NaCl 0,9 por ciento) en evaluación aguda (8,3 días), crónica (33,2 días) y recuperación del daño testicular (66.4 días). El arsénico reduce el número de espermatozoides en el tratamiento crónico (33,2 días) y este efecto continuó hasta 66,4 días. El efecto tóxico de As también altero la morfología de los espermatozoides en todos los períodos de tratamiento cuando se compara con el grupo control negativo. Sin embargo, metalonina indujo efectos protectores en períodos de 33,2 y 66,4 días de tratamiento. La estabilidad del ADN se vio afectada significativamente por el arsénico en todos los periodos, pero en el tratamiento crónico (33,2 días) con AsMe se observa un aumento de la estabilidad em comparación com el grupo tratado con arsénico. Sin embargo, la melatonina protege parcialmente a los espermatozoides del daño causado por arsénico, especialmente durante los períodos de 33,2 y 66,4 días.

Predictive factors influencing pregnancy rates after intrauterine insemination

So far, many studies investigated factors that affect pregnancy rates after intrauterine insemination [IUI]. Various investigators have not agreed on the nature and ranking of these criteria. The aim of this study was to assess the predictive factors for pregnancy rate after controlled ovarian hyperstimulation [COH]/ IUI. Retrospective study of all patients undergoing IUI at Zeynep Kamil Gynecologic and Pediatric Training and Research Hospital from January 2006 to December 2009. In total 980 IUI cycles in 569 couples were analyzed. All women in the study underwent ovarian stimulation using gonadotropin and IUI was performed 36 h after triggering ovulation. The primary outcome measure was clinical pregnancy rates. Predictive factors evaluated were female age, body mass index [BMI], duration of infertility, type of infertility, follicle stimulating hormone [FSH] level and estradiol [E[2]] on third day of the cycle, number of preovulatory follicles, endometrial thichness, total motil sperm [TMS] count, and ratio of progressive motile sperm. The overall clinical pregnancy rate was 4.7%. Among the predictive factors after multivariate logistic regression analysis level of BMI [<25 kg/m[2]], number of preovulatory follicles [> =2], level of FSH [<9.4 IU/L], level of E2 [<80 pg/ml] and the ratio of progressive motile sperm [>50%] significantly influenced the clinical pregnancy rate. Level of BMI, FSH, estradiol, number of preovulatory follicles and the ratio of progressive motile sperm may determine IUI procedure as optimum treatment model

Protective effect of melatonin on damage in the sperm parameters of mice exposed to diazinon / Efecto protector de la melatonina sobre el daño en los parámetros espermáticos de ratones expuestos a diazinón

Since normal sperm parameters can be altered by organophosphorous pesticides, this study intended to determine if melatonin is able to prevent the damage on sperm quality after an acute exposure to diazinon. Adult male mice were injected intraperitoneally with melatonin, diazinon (1/3 or 2/3 LD50) or both, and sperm parameters were evaluated on days 1 or 32 post injection. Groups treated with diazinon showed elevated lipid peroxidation levels on day 1 post treatment, while groups pretreated with melatonin before diazinon showed no difference compared to control. Sperm count showed a significant decrease in both diazinon-treated groups only on day 32 post injection; no differences were observed in groups pretreated with melatonin prior to diazinon compared to control. The percentage of abnormal sperm morphology increased in the diazinon-treated groups only on day 32 postinjection. The administration of melatonin prior to exposure to diazinon prevents the alteration of sperm parameters commonly caused by organophosphates, possibly due to its antioxidant properties.

Debido a que los parámetros normales de los espermatozoides pueden ser alterados por algunos contaminantes como los pesticidas organofosforados, este estudio pretende determinar si melatonina es capaz de prevenir o proteger del daño en la calidad espermática, después de una exposición aguda a diazinon. Ratones machos adultos fueron inyectados via intraperitoneal con diazinon 1/3 y 2/3 de la LD50 y otro grupo tratados con melatonina + 1/3 diazinon LD50 y melatonina + 2/3 LD50. Los parámetros espermáticos fueron evaluados al día 1 y al día 32 post tratamiento. Los grupos tratados con diazinon solo o conjugado con melatonina mostraron un incremento significativo en los niveles de lipoperoxidación en el tratamiento después de un día. Al día 32 no se observan diferencias significativas con el grupo control. El recuento espermático al día 1 no presenta diferencias entre los grupos tratados y el control. Sin embargo al día 32 los grupos tratados con diazinon solo, muestran una disminución significativa, solo el grupo de melatonina +1/3 diazinon, presenta valores similares al grupo control. La morfología espermática normal presenta una disminución significativa en grupos tratados con diazinon, pero un aumento significativo al día 32 en los grupos tratados con melatonina. Los mayores porcentajes de anormalidades se presentan en la cabeza y la cola de los espermatozoides. La administración de melatonina antes de la exposición al diazinon evita las alteraciones de los parámetros espermáticos, comúnmente causada por organofosforados, posiblemente debido a sus propiedades antioxidantes.

Identifying potential pitfalls in interpreting mitochondrial DNA mutations of male infertility cases.

Background & objectives: Recently, a significantly higher ratio of nucleotide changes in the mtDNA genes: COII, ATPase 6, ATPase 8, ND2, ND3, ND4, and ND5 was reported in spermatozoa from populations of infertile Indian men, compared suggesting that screening for mtDNA mutations could provide insight into the aetiology of male infertility. In this study, we examined the published data and found serious errors in the original acquisition and analysis of the data. Methods: The mtDNA data associated with male infertility in Indian populations were retrieved from the published sources. The mtDNA substitution values of infertile and control groups were evaluated using phylogenetic methods and previously published mtDNA phylogenies. Results: Most of the mtDNA polymorphisms reported as significantly correlated with infertility were more commonly found in general populations. Further, our analysis showed that some of the mtDNA substitutions were erroneously overestimated in the infertile groups and underestimated in the control groups, and vice-versa. Interpretation & conclusions: Contrary to earlier claims, our analysis demonstrated no significant association between the mtDNA polymorphisms and male infertility in these studies. Further, these errors in the published data impune the usefulness of mitochondrial molecular analyses in male infertility diagnosis.

Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa

The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.

Ejaculate characteristics of rabbits infected with Trypanosoma congolense and changes caused after treatment with Diminazene aceturate (Diminaveto®) / Características eyaculatorias de conejos infectados con Trypanosoma congolense y cambios causados después del tratamiento con Diminazeno aceturato (Diminaveto®)

The effects of experimental Trypanosoma congolense infection on the ejaculate of rabbits and changes caused after treatment with Diminaveto® were investigated using 24 New Zealand White rabbits (bucks). The bucks were housed singly in standard rabbit cages and fed on specialized ration containing 10 percent Protein supplement, grains, legume, salt and fresh water ad libitum during the study. Data on ejaculate characteristics were collected from all the bucks in the first phase (i.e. before infection) and in the second phase (i.e. during infection, with 4.8x10(5) Trypanosoma congolense, intraperitoneally). Similar data were collected from 12 randomly selected bucks treated with 7.0mg/kg Diminaveto® following reconstitution during the third phase. Data collected were analysed using the Paired T- Test and Analysis of Variance. The infection led to significant (P< 0.05) reduction in spermatozoa motility, concentration and mass activity, with a significant (P< 0.05) increase in percentage of sperm cells with morphological abnormalities. Treatment with Diminaveto® led to improvement in all ejaculate parameters investigated. However, it was observed that the ejaculate did not attain the "before-infection" status following treatment with Diminaveto®. The study showed that infection with Trypanosoma congolense in rabbits caused significant reduction in ejaculate characteristics. Treatment with Diminaveto® however led to improvement in the ejaculate though at a rate slower than that at which the infection caused the reduction.

Fue estudiado en 24 conejos Nuevo zelandeses blancos, machos, los efectos de la infección experimental de Trypanosoma congolense sobre la eyaculación y los cambios producidos después del tratamiento con Diminaveto® . Los machos fueron colocados individualmente en jaulas de conejos estándar. Durante el estudio fueron alimentados con ración especializada, con 10 por ciento de suplemento de proteínas, granos, legumbres, sal y agua fresca ad libitum. Los datos sobre las características de la eyaculación se obtuvieron de todos los machos en la primera fase (es decir, antes de la infección) y en la segunda fase (es decir, durante la infección, con 4,8x10(5) Trypanosoma congolense, por vía intraperitoneal). Datos similares se obtuvieron de 12 machos al seleccionados al azar, tratados con Diminaveto® 7,0mg/kg después de la reconstitución durante la tercera fase. Los datos fueron analizados mediante t de student y análisis de varianza. La infección fue significativa (P <0,05) habiendo reducción de la motilidad de los espermatozoides, la concentración y actividad de masas, con un efecto significativo (P <0,05) aumento en el porcentaje de espermatozoides con anomalías morfológicas. El tratamiento con Diminaveto® condujo a una mejoría en todos los parámetros investigados del eyaculado. Sin embargo, se observó que el eyaculado no alcanzó el "antes de la infección" tras el tratamiento con Diminaveto®. Además, el estudio mostró que la infección con Trypanosoma congolense en conejos causó una reducción significativa en las características del eyaculado. Sin embargo, el tratamiento con Diminaveto ® condujo a una mejoría en la eyaculación aunque a un ritmo más lento que en la infección causada por la reducción.

Rat spermatogenesis damage in intermittent hypobaric hypoxia and the protective role of melatonin. II: testicular parameters / Daño de la espermatogénesis en rata en la hipoxia hipobárica intermitente y rol protector de la melatonina. II: parámetros testiculares

At present it is not clear if male fertility is affected by intermittent hypobaric hypoxia (IHH). This is an important issue since a large human population works over 3000 masl. This study analyzes testicular changes in adult Sprague Dawley rats after five cycles of IHH (7 day exposure to 4200 masl in a hypobaric chamber / 7 day at 500 masl). The animals were separated into groups of 8, one group was exposed to hypoxia (7 days), and the others to IHH for one to five cycles. Controls (500 masl) were examined at the beginning and at the end of the 70 experimental days. A duplicate set of rats treated with melatonin (supposedly protecting from hypoxia) was also examined, as were their controls, injected with 0,03 percent ethanol (melatonin solvent).Immunohistochemical and histometric analysis of testicular tissue were performed. Damage caused by IHH increases with time. Morphometry reveals an increase in tubular and luminal diameters and a reduction in epithelial height. Inmunohistochemistry for HIF-1alpha shows an increase with time, however the opposite happens with HSP-70. Spermatogenic cells submitted to comet assay present an increase of (+) cells. Melatonin counteracts all this damage, possibly due to its high efficiency as a reactive oxygen species scavenger. In conclusion, IHH exposure damages male reproductive function.

Actualmente no se conoce claramente si la fertilidad masculina se afecta por la hipoxia hipobarica intermitente (IHH). Esto es de importancia porque una gran población humana trabaja sobre 3000 metros sobre el nivel del mar (sml). Este trabajo analiza los cambios testiculares en ratas adultas Sprague Dawley luego de cinco ciclos de IHH (7 días a 4.200 sml) en una camara hipobarica/7días a 500 metros: Normoxia). Los animales de dividieron en grupos de 8; un grupo expuesto a hipoxia (7 días ) y los otros a IHH por uno y hasta cinco ciclos. Los controles ( 500 sml) se analizaron al inicio y al final de los 70 días experimentales. Un set duplicado de ratas tratadas con melatonina (considerada protectora de la Hipoxia) se examinó también, así como sus controles, inyectados con etanol 0,03 por ciento (solvente de la melatonina).Se realizó análisis histométrico e inmunohistoquímico del tejido testicular. El daño causado por IHH aumenta con el tiempo. La morfometría reveló un aumento de los diámetros del túbulo y lumen y una reducción de la altura del epitelio. La inmunohistoquímica de HIF-1 alpha muestra aumento del número de túbulos positivos con el tiempo aunque lo opuesto ocurre para HSP-70. El ensayo de cometa muestra un aumento del número de células espermátogénicas (+). La melatonina controla este daño, posiblemente debido a asu alta eficiencia como neutralizador de especies reactivas del oxígeno. En conclusión, la exposición a IHH daña la función reproductiva masculina.

Rat spermatogenesis damage in intermittent hypobaric hypoxia and the protective role of melatonin: I cauda epididymal spermatozoa / Daño en la espermatogénesis de rata en hipoxia hipobárica intermitente y el papel protector de la melatonina: I espermatozoides de la cauda epididimaria

At present it is not clear if male fertility is affected by intermittent hypobaric hypoxia (IHH). This is an important issue since a large human population works over 3000 masl. This study analyzes epididymal sperm, in adult Sprague Dawley rats after five cycles of IHH (7 days exposure to 4200 masl in a hypobaric chamber / 7 days at 500 masl). The animals were separated into groups of 8, one group was exposed to hypoxia (7 days), and the others to IHH for one to five cycles. Controls (500 masl) were examined at the beginning and at the end of the 70 experimental days. A duplicate set of rats treated with melatonin (supposedly protecting from hypoxia) was also examined, as were their controls, injected with 0,03 percent ethanol (melatonin solvent). Epididymal sperm parameters, were evaluated. Damage caused by IHH increases with time. Sperm counts drop, while sperm chromatin swelling, DNA instability (metachromasia with acridine orange epifluorescence) and comet (+) tests increase. Melatonin counteracts all this damage, possibly due to its high efficiency as a reactive oxygen species scavenger. In conclusion, IHH exposure damages sperm quality and therefore male reproductive function.

En la actualidad no está claro si la fertilidad masculina se ve afectada por la hipoxia hipobárica intermitente (HHI). Esta es una cuestión importante, ya que una gran población humana trabaja a más de 3000 metros sobre el nivel del mar (msnm). Este estudio analiza los espermatozoides del epidídimo, en ratas Sprague Dawley adultas, después de cinco ciclos de HHI (7 días de exposición a 4200 msnm en una cámara hipobárica / 7 días a 500 msnm). Los animales fueron separados en grupos de 8, un grupo fue expuesto a la hipoxia (7 días), y los otros a HHI de uno a cinco ciclos. Los controles (500 msnm) se examinaron al comienzo y al final de los 70 días de experimentación. Un grupo duplicado de ratas tratadas con melatonina (supuestamente protegiendo de la hipoxia) también fue examinado, al igual que los controles, inyectados con 0,03 por ciento de etanol (solvente de melatonina). Los parámetros espermáticos del epidídimo fueron evaluados. Los daños causados por la HHI aumentaron con el tiempo. Se evaluó el conteo espermático en gota, mientras la cromatina espermática esta hinchada, la inestabilidad del ADN (metacromasia epifluorescente con naranjo de acridina) y la prueba de aumento (+) de cometas. La melatonina neutralizó todo el daño, posiblemente debido a su alta eficacia como un eliminador de especies reactivas de oxígeno. En conclusión, la exposición a HHI daña la calidad espermática y por lo tanto la función reproductora masculina.

Oxidative stress and sperm mitochondrial DNA mutation in idiopathic oligoasthenozoospermic men.

Physiological function of reactive oxygen species (ROS) has been known since a long, but recently toxic effects of ROS on spermatozoa have gained much importance in male infertility. Mitochondrial DNA (mtDNA) is believed to be both source and target of ROS. mtDNA unlike nuclear DNA is not compactly packed and hence more susceptible to oxidative stress (OS) than nuclear DNA. In the present study, the role of OS in mitochondrial genome changes was studied in men with idiopathic infertility. The study included 33 infertile oligo-asthenozoospermic (OA) men and 30 fertile controls. Semen analyses were performed and OS was measured by estimating the level of malondialdehye (MDA) in the seminal plasma and ROS in the sperm. Sperm mtDNA was sequenced by standard PCR-DNA sequencing protocol for ATPase and nicotinamide adenine dinucleotide dehydrogenase (ND) groups of genes. Sperm count and progressive motility were found to be significantly lower in infertile group than the fertile controls. Semen MDA and ROS levels of infertile group were significantly higher (p<0.0001), when compared to the control group. However, catalase and glutathione peroxidase (GPx) levels were significantly lower in infertile group, compared to controls, but no significant difference in superoxide dismutase (SOD) activity was observed between control and cases. This might be due to higher expression of SOD alone in order to overcome OS in the semen. mtDNA analysis showed significant and high frequency of nucleotide changes in the ATPase (6 and 8), ND (2, 3, 4 and 5) genes of infertile cases compared to the controls. Hence excess ROS and low antioxidant levels in the semen might cause mtDNA mutations and vice versa in OA men that might impair the fertilizing capacity of spermatozoa. Thus, it is important to understand the etiology of mitochondrial genome mutations in idiopathic OA cases for better diagnostic and prognostic value in infertility treatment/assisted reproductive technique

Effect of emotional stress on sperm quality.

BACKGROUND & OBJECTIVE: Emotional stress plays a detrimental role on fertility. In this study male patients with idiopathic infertility were selected after evaluation of psychological stress to evaluate a positive effect of a stress therapy on their semen quality. METHODS: A total of 20 patients with infertility were enrolled in the study and randomly divided in two groups. Ejaculates were examined by light and transmission electron microscopy (TEM). Meiotic segregation was also investigated by fluorescence in situ hybridization (FISH). Ten patients were treated with Conveyer of Modulating Radiance (CRM) therapy and sperm characteristics and meiotic segregation were evaluated again three months at the end of treatment. RESULTS: TEM data showed that, among sperm pathologies, necrosis and apoptosis were higher and the number of "healthy" sperm was significantly reduced in both groups of stressed men compared to reference values. The number of "healthy" sperm was significantly higher in the treated group after therapy, indicating a recovery of sperm quality, although no significant decrease in sperm pathologies was observed. FISH analysis showed that the mean frequencies of sex chromosomes disomies and diploidies significantly decreased after stress therapy. INTERPRETATION & CONCLUSION: The effects induced by stress also seem to include meiotic and structural alterations in sperm cells. The spermatogenic process was improved after a cycle of CRM therapy indicating that stress is an additional risk factor for idiopathic infertility.

Clinical relevance of oxidative stress and sperm chromatin damage in male infertility: an evidence based analysis

Oxidative stress (OS) in the reproductive tract is now a real entity and concern due to the potential harmful effects of high levels of reactive oxygen species (ROS) on sperm number, motility, quality, and function including damage to sperm nuclear DNA. Evaluation of OS related damage to non-functional sperm is highly relevant as intracytoplasmic sperm injection (ICSI) technique, an effective therapy for severe male factor infertility, bypasses the majority of reproductive tract deficiencies. Despite the controversial findings in the existing literature, there is now enough evidence to show that sperm DNA damage is detrimental to reproductive outcomes. In addition, spermatozoa of infertile men are suggested to carry more DNA damage than do the spermatozoa from fertile men. Besides impairment of fertility such damage is likely to increase the transmission of genetic diseases during the assisted reproductive procedures. Standardization of protocols to assess reactive oxygen species and DNA damage is very important in introducing these tests in such clinical practice. Thus evaluation of seminal ROS levels and extent of sperm DNA damage especially in an infertile male may help develop new therapeutic strategies and improve success of assisted reproductive techniques (ART).