Experimental mild increase in testicular temperature has drastic, but reversible, effect on sperm aneuploidy in men: A pilot study.

In mammals testicular and epididymal temperature increase impairs spermatogenesis. This experimental study investigates the effects of a mild testis temperature increase (i.e. testis temperature remains below core body temperature) on sperm aneuploidy in men. In 5 fertile volunteers a testicular temperature increase was induced by maintaining the testes at suprascrotal position using specially designed underwear for 15 ±â€¯1 h daily for 120 consecutive days. After heating men were followed for next 180 days. A control group (27 men) was recruited. Semen samples were collected before, during and after heating period and analyzed for chromosomes X, Y and 18 for aneuploidy using FISH. A total of 234,038 spermatozoa were studied by FISH. At day 34 of heating, mean sperm aneuploidy values were not modified. From day 34 of heating until day 45 post heating, FISH evaluation was not possible due to the drastic fall of sperm count. At day 45 post-heating total sperm aneuploidy percentage was twice higher than before heating whereas. Sex disomy (sperm XY18), sex chromosome nullisomy (sperm 18) were significantly higher than controls. These effects were completely reversed at 180 days post heat exposure. Conclusion: A mild rise in testicular temperature significantly increases sperm aneuploidies, reflecting an effect on the meiosis stage of spermatogenesis. The effect of heating was reversible and suggests that recovery of aneuploidy to normal values requires at least two cycles of spermatogenesis. Nonetheless, the low number of volunteers was a limitation of this pilot study and warrants further research on larger population.

Mild experimental increase in testis and epididymis temperature in men: effects on sperm morphology according to spermatogenesis stages.

BACKGROUND: A mild increase in testicular and epididymal temperatures in men, bulls and rams (pendulous scrotum) inhibits spermatogenesis and increases the percentage of sperm with an abnormal morphology. However, the stages of spermatogenesis that are most sensitive to a mild increase in testicular temperature in men are unknown. The aim of the present study was to explore the effects of a mild induced increase in testicular and epididymal temperature (i.e., testicular temperature maintained below the core body temperature) on sperm morphology in humans depending on the physiological time of spermatogenesis and epididymal transit. METHODS: Five healthy volunteers were enrolled in an experimental study in which testicular and epididymal temperatures were increased by maintaining the testes in a supra-scrotal position with a specially designed underwear worn 15±1 h a day for 120 consecutive days. Semen collection was scheduled on specific days depending on spermatogenic stages and epididymal transit. RESULTS: Sperm morphology and the multiple anomalies index (MAI) were analysed before, during and after heating. This mild induced increase in testicular and epididymal temperatures resulted in a significant rise in the percentage of morphologically abnormal spermatozoa on day 34 of heating, which remained elevated throughout the heating period and persisted until 45 days after cessation of heating. The MAI was significantly increased on day 20 throughout the heating period and persisted 45 days after cessation of heating. An increase in the percentage of anomalies in the sperm head, acrosome or tail occurred on days 34 and/or 45 of heating. Abnormal sperm morphology and MAI reverted to control values 73 days after cessation of heating. CONCLUSIONS: A mild sustained increase in testicular and epididymal temperature in man leads morphological abnormalities in spermatozoa mainly due to an impairment of spermiogenesis and meiosis.

Sperm cryopreservation incidence in men with testicular cancer: towards a stabilization in testicular cancer incidence? Results from the CECOS network.

BACKGROUND: Testicular cancer (TC) represents 1% of all new male cancer cases but remains the most frequent cancer in adolescents and young adults in industrialized countries. In this study, we assessed time trends in use of sperm cryopreservation by men with TC from 1990 to 2013 in France. METHODS: We collected data from patients diagnosed with TC who underwent sperm cryopreservation in the French national network of sperm banks. Trends in the incidence of sperm cryopreservation were estimated through two statistical models: the commonly used Poisson regression model and the Verhulst model. RESULTS: Between 1990 and 2013, the overall incidence of sperm cryopreservation rose from 1.73 to 5.57 per 100,000 person-years. Poisson regression predicted an incidence of 9 per 100,000 [95% CI = 8.66-9.34] in 2020. However, since 2005, the observed sperm cryopreservation rate seems to be attenuating. The Verhulst model predicted an incidence of 6 per 100,000 after 2020. CONCLUSIONS: Limitations include the impossibility of analyzing age-standardized incidence. Based on the Verhulst model, results suggest that it is still relevant to follow up TC incidence and sperm cryopreservation in order to confirm or refute the potential decrease already observed in this disease.

CONTEXTE: Le cancer des testicules (CT) représente environ 1% de l'ensemble des cas de cancer chez les hommes, mais il demeure néanmoins le cancer le plus fréquent chez les adolescents et les jeunes adultes dans les pays industrialisés. Dans cette étude, nous avons évalué les variations temporelles des autoconservations de sperme réalisées par les hommes atteints d'un CT entre 1990 et 2013 en France. MÉTHODES: Les données proviennent des autoconservations de sperme réalisées auprès de patients diagnostiqués avec CT, issues du réseau national français des banques de sperme. Les tendances de l'incidence des autoconservations de sperme ont été estimées à l'aide de deux modèles statistiques: la régression de Poisson, couramment utilisée, et le modèle de Verhulst. RÉSULTATS: Entre 1990 et 2013, l'incidence globale des autoconservations de sperme est. passée de 1,73 à 5,57 pour 100,000 personnes-années. La régression de Poisson montre une estimation de l'incidence de près de 9 pour 100,000 [IC à 95% = 8,66-9,34] en 2020. Cependant, depuis 2005, le taux observé des autoconservations de sperme semble s'atténuer. Le modèle de Verhulst estime alors une incidence aux alentours de 6 pour 100,000 après 2020. CONCLUSIONS: Les limites de cette étude comprennent l'impossibilité d'analyser l'incidence standardisée sur l'âge. Sur la base du modèle de Verhulst, les résultats suggèrent qu'il est. toujours et encore pertinent d'étudier l'évolution de l'incidence du cancer du testicule et des autoconservations de sperme afin de confirmer ou non la diminution/stagnation potentielle déjà observée dans cette maladie.

Sperm aneuploidy and DNA fragmentation in unexplained recurrent pregnancy loss: a multicenter case-control study.

BACKGROUND: Recurrent pregnancy loss (RPL) is defined as the loss of at least three pregnancies in the first trimester. Although the most common cause is embryo aneuploidy, and despite female checkup and couple karyotyping, in about 50% of cases RPL remain unexplained. Male implication has little been investigated and results are discordant. In this context, we conducted a multi-center prospective case-control study to investigate male gamete implication in unexplained RPL. METHODS: A total of 33 cases and 27 controls were included from three university hospitals. We investigated environmental and family factors with a detailed questionnaire and andrological examination, sperm characteristics, sperm DNA/chromatin status using the sperm chromatin structure assay (SCSA) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and sperm aneuploidy using fluorescence in situ hybridization (FISH). The Mann-Whitney test and the Wilcoxon or Fisher exact tests were used. A non-parametric Spearman correlation was performed in order to analyze the relationship between various sperm parameters and FISH and sperm DNA fragmentation results. RESULTS: We found significant differences between cases and controls in time to conceive, body mass index (BMI), family history of infertility and living environment. In cases, total sperm motility and the percentage of morphologically normal spermatozoa were significantly decreased. No difference was found between cases and controls in sperm DNA fragmentation or chromatin integrity. In cases, spermatozoa with aneuploidy, hyperhaploidy and chromosome 18 disomy were significantly increased. CONCLUSIONS: This prospective case-control study is one of the largest to examine environmental factors, sperm characteristics, sperm DNA fragmentation and chromatin, and chromosome anomalies in spermatozoa in relation to unexplained recurrent pregnancy loss. The originality of our study lies in the comprehensive andrological examination and search for risk factors and fertility history. Further studies are needed to confirm the links between unexplained RPL and a male family history of infertility or miscarriages. The increased sperm aneuploidy observed in unexplained RPL supports a male etiology. These data pave the way for further studies to demonstrate the value of preimplantation genetic screening in men with increased sperm aneuploidy whose partners experience unexplained RPL.

CONTEXTE: Les fausses couches à répétition (FCR) sont définies lorsqu'au moins trois fausses couches ont eu lieu au cours du premier trimestre. Bien que la cause la plus fréquente soit l'aneuploïdie embryonnaire, et malgré un bilan chez la femme et un caryotype du couple, dans environ 50% des cas, les FCR restent inexpliquées. L'implication masculine a été peu étudiée et les résultats restent discordants. Ainsi, nous avons réalisé une étude cas-témoins prospective et multicentrique afin d'investiguer l'implication du gamète mâle dans les FCR inexpliquées. MÉTHODES: Un total de 33 cas et de 27 témoins ont été inclus recrutés au sein de trois hôpitaux universitaires. Nous avons étudié les facteurs environnementaux et familiaux à partir d'un questionnaire détaillé ainsi que les données de l'examen andrologique, les caractéristiques du sperme, la fragmentation de l'ADN et la chromatine du spermatozoïde en utilisant le sperm chromatine structure assay (SCSA) et le test du TUNEL, ainsi que l'aneuploïdie des spermatozoïdes grâce à la méthode d'hybridation in situ de sonde chromosomique (FISH). Le test de Mann-Whitney et les tests exacts de Wilcoxon ou de Fisher ont été utilisés. Une corrélation de Spearman non-paramétrique a été réalisée afin d'analyser la relation entre les divers paramètres de sperme et les résultats de fragmentation d'ADN du sperme et les résultats de la FISH. RÉSULTATS: Nous avons trouvé des différences significatives entre les cas et les témoins pour le délai de conception, l'indice de masse corporelle (IMC), les antécédents familiaux d'infertilité et le milieu de vie. Chez les cas, la mobilité totale des spermatozoïdes et le pourcentage de spermatozoïdes normaux étaient significativement diminués. Aucune différence n'a été trouvée entre les cas et les témoins concernant la fragmentation de l'ADN des spermatozoïdes ou l'intégrité de la chromatine. Chez les cas, la fréquence des spermatozoïdes présentant une aneuploïdie, une hyperhaploïdie et une disomie du chromosome 18 étaient significativement augmentée. CONCLUSIONS: Cette étude cas-témoins prospective est. l'une des plus importantes ayant investigué à la fois les facteurs environnementaux, les caractéristiques des spermatozoïdes, la fragmentation et la chromatine de l'ADN des spermatozoïdes et les anomalies chromosomiques des spermatozoïdes en rapport avec les fausses couches à répétition inexpliquée. L'originalité de notre étude réside dans l'examen andrologique complet et la recherche des facteurs de risque et des antécédents reproductifs. D'autres études sont nécessaires pour confirmer les liens entre les FCR inexpliquées et les antécédents familiaux masculins d'infertilité ou de fausses couches. L'augmentation de l'aneuploïdie des spermatozoïdes observée chez les cas présentant des FCR inexpliquées plaide en faveur d'une étiologie masculine. Ces données ouvrent la voie à d'autres études pour démontrer l'utilité d'un dépistage génétique préimplantatoire chez les hommes présentant une augmentation de l'aneuploïdie des spermatozoïdes dont les partenaires présentent des FCR inexpliquées.

Patterns of residual HIV-1 RNA shedding in the seminal plasma of patients on effective antiretroviral therapy.

BACKGROUND: More and more HIV-1-infected men on effective antiretroviral treatment (ART) have unprotected sex in order to procreate. The main factor influencing transmission is seminal HIV shedding. While the risk of HIV transmission is very low, it is difficult to assess in individuals. Nevertheless, it should be quantified. RESULTS: We retrospectively analysed seminal plasma HIV-1 shedding by 362 treated HIV-infected men attending a medically assisted reproduction centre (1998-2013) in order to determine its frequency, the impact of the antiretroviral regimen on HIV shedding, and to identify shedding patterns. The HIV-1 virus loads in 1396 synchronized blood and semen samples were measured, and antiretroviral treatment, biological and epidemiological data were recorded. We detected isolated HIV-1 shedding into the seminal plasma in 5.3% of patients on efficient antiretroviral treatment, but there was no association with the HIV antiretroviral drug regimen or the CD4 cell count. These men had undergone more regimen changes since treatment initiation and had been on the ongoing drug regimen longer than the non-shedding men. The patterns of HIV seminal shedding among patients with undetectable HIV blood virus load varied greatly. HIV seminal shedding can occur as long as 5 years after starting antiretroviral treatment. CONCLUSIONS: The seminal HIV load was used to monitor risk for infertile HIV-infected patients on an assisted reproductive technology program. This can still be recommended for patients who recently (6 months) started ART, or those with a poor history of adherence to ART but may also be usefull for some patients during counselling. Residual HIV seminal shedding is probably linked to breaks in adherence to antiretroviral treatment but local genital factors cannot be ruled out.

RÉSUMÉ: De plus en plus d'hommes sous traitement antirétroviral (ART) ont des rapports sexuels non protégés à des fins de procréation. Le principal déterminant de la transmission sexuelle est. l'excrétion séminale du VIH. Malgré un risque de transmission très faible, il reste difficile à évaluer au niveau individuel. Dans ce contexte, l'étude de l'excrétion séminale du VIH, notamment chez des hommes sous traitement antirétroviral, est. d'un grand intérêt. RÉSULTATS: Nous avons analysé rétrospectivement l'excrétion séminale du HIV chez 362 hommes sous traitement antirétroviral consultant un centre d'assistance médicale à la procréation (1998­2013) pour déterminer sa fréquence, l'impact des antirétroviraux sur l'excrétion du HIV et identifier les profils d'excrétion. Les charges virales HIV-1 ont été mesurées dans 1396 échantillons de sang et de sperme prélevés concomitamment et les traitements, les données biologiques et épidémiologiques recueillis. Nous avons détecté une excrétion dans le plasma séminal isolée chez 5,3% des patients sous traitement antirétroviral efficace mais nous n'avons pas trouvé d'association avec la composition du traitement antirétroviral ou le taux de lymphocytes T CD4+. Ces hommes avaient eu plus de changements thérapeutiques et leur traitement avait été instauré depuis plus longtemps que pour les hommes non excréteurs. Les profils d'excrétion séminale du HIV parmi les patients avec une charge virale indétectable dans le sang étaient très variables. L'excrétion séminale du HIV peut survenir jusqu'à 5 ans après l'instauration du premier traitement antirétroviral. CONCLUSIONS: La charge virale séminale du HIV était l'outil classique d'évaluation du risque viral de transmission pour les patients infertiles infectés par HIV et inclus dans les programmes d'assistance médicale à la procréation. Ceci peut continuer à être recommandé chez les patients ayant débuté un traitement antirétroviral dans les 6 mois précédent ou chez ceux avec des antécédents de mauvaise adhérence au traitement mais peut aussi être utile pour le conseil de certains patients. Le risque résiduel d'excrétion séminale du HIV est. probablement lié à des défauts d'adhérence au traitement antirétroviral mais des facteurs génitaux ne peuvent pas être éliminés.

Effect of acute Zika virus infection on sperm and virus clearance in body fluids: a prospective observational study.

BACKGROUND: Evidence of human sexual transmission during Zika virus emergence is a matter of concern, particularly in procreation, but to date, kinetics of seminal shedding and the effects of infection on human reproductive function have not been described. To investigate the effects of Zika virus infection on semen and clearance of Zika virus from semen and body fluids, we aimed to study a cohort of Zika virus-infected men. METHODS: This prospective observational study recruited men presenting with acute Zika virus infection at Pointe-à-Pitre University Hospital in Guadeloupe, French Caribbean, where a Zika virus outbreak occurred between April and November, 2016. Blood, urine, and semen were collected at days 7, 11, 20, 30, 60, 90, and 120 after symptom onset, and semen characteristics, such as total sperm count, sperm motility, vitality, and morphology, and reproductive hormone concentrations, such as testosterone, inhibin, follicle-stimulating hormone, and luteinising hormone, were assessed. At days 7, 11, and 20, semen was processed to isolate motile spermatozoa. Zika virus RNA was detected by RT-PCR using whole blood, serum, urine, seminal plasma, semen cells, and motile spermatozoa fractions. Zika virus was isolated from different sperm fractions on Vero E6 cultures. FINDINGS: 15 male volunteers (mean age 35 years [SD 5; range 25-44) with acute Zika virus infection and positive Zika virus RNA detection in blood or urine were enrolled. Total sperm count was decreased from median 119 × 106 spermatozoa (IQR 22-234) at day 7 to 45·2 × 106 (16·5-89·6) at day 30 and 70 × 106 (28·5-81·4) at day 60, respectively, after Zika virus infection. Inhibin values increased from 93·5 pg/mL (IQR 55-162) at day 7 to 150 pg/mL (78-209) at day 120 when total sperm count recovered. In motile spermatozoa obtained after density gradient separation, Zika virus RNA was found in three of 14 patients at day 7, four of 15 at day 11, and four of 15 at day 20, and replication-competent virus was found in the tested patient. Seminal shedding kinetics seemed heterogeneous among patients. Whole blood was the fluid most frequently positive for Zika virus RNA (62 of 92 samples) and three patients remained positive at day 120. INTERPRETATION: Semen alterations early after acute Zika virus infection might affect fertility and could be explained by virus effects on the testis and epididymis. Frequency of shedding and high viral load in semen, together with the presence of replicative virus in a motile spermatozoa fraction, can lead to Zika virus transmission during sexual contact and assisted reproduction procedures. Whole blood seems to be the best specimen for Zika virus RNA detection, diagnosis, and follow-up. FUNDING: Agence de la Biomédecine/Agence Régionale de Santé de la Guadeloupe/Inserm-REACTing.

Diagnosis of Partial Retrograde Ejaculation in Non-Azoospermic Infertile Men with Low Semen Volume.

In non-azoospermic patients with low semen volume (LSV), looking for partial retrograde ejaculation (PRE) by searching sperm in the postejaculatory urine (PEU) is required. The use of a retro-ejaculatory index (R-ratio) was suggested to define PRE, but none of the studies indicated a specific threshold above which PRE must be considered. Our objective was to propose a threshold value for the R-ratio as indicative of PRE in patients with LSV selected to be devoid of any known causes or risk factors for retrograde ejaculation or LSV. Among our data base (2000-2009) including 632 patients with PEU, 245 male patients from infertile couples who had had a first semen analysis with LSV (< 2mL) and a second semen analysis associated with PEU, were selected on the previous criteria. A prospective control group was randomly constituted (2007-2008) of 162 first consulting male patients from infertile couples, with a normal semen volume (≥ 2mL) on a first semen analysis and who accepted to collect PEU with their usual second semen analysis, selected on the previous criteria. To define an R-ratio threshold indicative of PRE, we used a ROC curve analysis and a regression tree based on a classification and regression tree (CART) algorithm. Of the 245 LSV patients, 146 still presented low semen volume (< 2 mL) on the second semen analysis. From the use of the CART algorithm, two low (1.5% and 2.8%) and two high R-values (7.1% and 8.3%) were defined, according to the lower reference limit for semen volume of 2.0 mL (WHO 1999) or 1.5 mL (WHO 2010) respectively. As only one or no patient with normal semen volume was observed above the two high R-values, we suggest an R-value higher than the range of [7.1-8.3]% as indicative of PRE until confirmation by a prospective multicenter study.

Sperm aneuploidy after testicular cancer treatment: data from a prospective multicenter study performed within the French Centre d'Étude et de Conservation des Oeufs et du Sperme network.

OBJECTIVE: To study sperm aneuploidy in a population of testicular cancer (TC) patients treated with the use of either bleomycin-etoposide-cisplatin (BEP) chemotherapy or radiotherapy. DESIGN: Multicenter prospective longitudinal study of TC patients analyzed before treatment and after 3, 6, 12, and 24 months (T3-T24). PATIENT(S): Fifty-four TC patients and a control group of 10 fertile sperm donors. SETTING: University hospital laboratories. INTERVENTION(S): Routine semen analyses; sperm aneuploidy and diploidy. MAIN OUTCOME MEASURE(S): Comparison of sperm characteristics and sperm chromosome abnormalities during TC patient follow-up. RESULT(S): Semen characteristics recovered pretreatment values 12 months after radiotherapy and 24 months after more than two BEP cycles. A significant increase in sperm disomy YY and XX was observed in the TC group before treatment compared with the control group. After more than two BEP cycles, the mean sperm aneuploidy rate increased significantly at T12 and reached the pretreatment value at T24. After radiotherapy, the mean sperm aneuploidy returned to the pretreatment value at T12. At T24, nearly 40% of TC patients did not recover their pretreatment sperm aneuploidy rate. CONCLUSION(S): Genetic counseling of TC patients should include information on the potential elevated risk of aneuploid conceptus from sperm recovered after treatment and the necessity to postpone conception up to ≥12 months after radiotherapy and ≥24 months after more than two BEP chemotherapy cycles. However, few men receiving one or two BEP cycles and some dropouts are the main limitations of this study.

Impact of Hodgkin or non-Hodgkin lymphoma and their treatments on sperm aneuploidy: a prospective study by the French CECOS network.

OBJECTIVE: To assess sperm production and aneuploidy in Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL) before and after treatments. DESIGN: Multicenter, prospective, longitudinal study of lymphoma patients analyzed before treatment and after 3, 6, 12, and 24 months. SETTING: University hospitals. PATIENT(S): Forty-five HL and 13 NHL patients were investigated before and after treatment. Treatment regimens were classified in two groups: ABVD (doxorubicin, bleomycin, vinblastine, dacarbazine) with or without (±) radiotherapy, and CHOP (doxorubicin, cyclophosphamide, vincristine, prednisone)/MOPP-ABV (mechlorethamine, oncovin, procarbazine, prednisone-doxorubicin, bleomycin, vinblastine). A control group of 29 healthy men was also studied. INTERVENTION(S): Semen analyses and aneuploidy study by FISH were performed at each time point. MAIN OUTCOME MEASURE(S): Comparison of mean sperm characteristics and percentage of sperm aneuploidy rates before and after treatment. RESULT(S): Before treatment, HL and NHL men had altered semen characteristics and higher sperm aneuploidy rates (median 0.76 [interquartile range 0.56-0.64]) than the control group (0.54 [0.46-0.74]). After treatment, sperm production was significantly lowered 3 and 6 months after ABVD ± radiotherapy or CHOP/MOPP-ABV. After ABVD ± radiotherapy, the aneuploidy rate increased significantly only at 3 months, and values obtained 1 or 2 years later were lower than pretreatment values. In contrast, in the CHOP/MOPP-ABV treatment group, semen characteristics and aneuploidy rate did not return to normal levels until 2 years after treatment. CONCLUSION(S): Lymphoma itself has consequences on sperm aneuploidy frequency before treatment. Moreover, lymphoma treatments have deleterious effects on sperm chromosomes related to treatment type and time since treatment. Patient counseling is essential concerning the transient but significant sperm aneuploidy induced by lymphoma and its treatments.

Sperm cryopreservation in adolescents and young adults with cancer: results of the French national sperm banking network (CECOS).

OBJECTIVE: To determine the feasibility of fertility preservation in adolescent males with cancer. DESIGN: Large multicenter retrospective study of male patients ≤20 years from 23 centers of a national network of sperm banks over a 34-year period. SETTING: Sperm banks. PATIENT(S): A total of 4,345 boys and young men aged 11 to 20 years. INTERVENTION(S): Age, cancer diagnosis, feasibility of sperm banking, and sperm parameters. MAIN OUTCOME MEASURE(S): Description of patients, and success of their fertility preservation. RESULT(S): We observed a mean yearly increase in referred patients of 9.5% (95% confidence interval, 9.1%-9.8%) between 1973 and 2007. Over the study period, the percentage of younger cancer patients who banked their sperm increased, especially in the 11-14 year age group, rising from 1% in 1986 to 9% in 2006. We found that 4,314 patients attempted to produce a semen sample, 4,004 succeeded, and sperm was banked for 3,616. The mean total sperm count was 61.75 × 10(6) for the 11-14 year age group, and 138.81 × 10(6) for the 18-20 year age group. It was noteworthy that intercenter variations in practices involving young patients seeking to preserve their fertility before cancer therapy were observed within this national network. CONCLUSION(S): Our results emphasize the need for decisive changes in public health policy to facilitate the access to reproductive health-care for young cancer patients.

Impact of lymphoma treatments on spermatogenesis and sperm deoxyribonucleic acid: a multicenter prospective study from the CECOS network.

OBJECTIVE: To determine consequences of lymphoma treatments on sperm characteristics and sperm DNA, and to evaluate predictors of sperm recovery. DESIGN: Multicenter prospective longitudinal study of patients analyzed before treatment and after 3, 6, 12, and 24 months. SETTING: University hospitals. PATIENT(S): Seventy-five Hodgkin lymphoma and non-Hodgkin lymphoma patients and a control group of 257 fertile men. INTERVENTION(S): Semen analyses, and sperm DNA and chromatin assessments. MAIN OUTCOME MEASURE(S): Comparisons of sperm characteristics before and after treatment. RESULT(S): Patients already had altered sperm characteristics before lymphoma treatment, with no identified risk factor. Sperm count, total sperm count, motility, and vitality decreased after treatment, with lowest values at 3 and 6 months. Twelve months after treatment, mean sperm count recovered to pretreatment values after doxorubicin, bleomycin, vinblastine, darcarbacine (ABVD) or ABVD+radiotherapy, but not after doxorubicin, cyclophosphamide, vincristine, prednisone (CHOP) or mechlorethamine, oncovin, procarbazine, prednisone (MOPP) chemotherapies. It was noteworthy that 7% of patients remained azoospermic at 24 months. After 24 months, Kaplan-Meier estimates showed that more than 90% of patients will recover normal sperm count after ABVD or ABVD+radiotherapy vs. 61% for CHOP chemotherapies. In multivariate analyses including diagnosis and treatment protocol, only pretreatment total sperm count was related to recovery. Compared with a control group, lymphoma patients had higher sperm chromatin alterations and DNA fragmentation before any treatment. After treatment, DNA fragmentation assessed by TUNEL assay and sperm chromatin structure assay decreased from 3 and 6 months, respectively, while remaining higher than in the control group during follow-up. CONCLUSION(S): Lymphoma patients had altered sperm DNA and chromatin before treatment. Lymphoma treatment had damaging effects on spermatogenesis. These data on both the recovery period according to treatment modalities and the pre- and post-treatment chromatin status of sperm are useful tools for counseling patients wishing to conceive.

A prospective study in male recipients of kidney transplantation reveals divergent patterns for inhibin B and testosterone secretions.

BACKGROUND: Male patients with chronic kidney disease often exhibit the biological and clinical hallmarks of an abnormal hypothalamo-pituitary-gonadal axis. It is known that dialysis does not reverse this impaired endocrine status; however, the impact of kidney transplantation (KT) is still controversial. The aim of our study was to investigate the levels of serum gonadotropins, testosterone, and inhibin B during dialysis and after KT. METHODS: A longitudinal and prospective single center study was led in an academic setting. Blood hormones levels were assayed by immunoassays in 53 men (mean age: 37 years) receiving dialysis (T0) and at 6 months post-KT (T180). These data were compared with those from 46 fertile semen donors (mean age: 37 years). The main outcome measure was the between-groups differences in hormones levels. A second criterion was the comparison of T0 and T180 hormones levels according to the immunosuppressive regimen. RESULTS: For patients ongoing dialysis, luteinizing hormone (LH) and follicle-stimulating hormone (FSH) mean levels were high, whereas testosterone and inhibin B mean levels remained normal. After KT, LH levels returned to normal whereas FSH was significantly increased. Testosterone levels remained normal whereas inhibin B levels significantly decreased. We found that the combination tacrolimus plus mycophenolic acid significantly decreased post-KT inhibin B levels. Moreover, we found that pre-graft inhibin-B level was independent of testosterone and could predict low post-operative inhibin B level with a sensitivity of 77% and a specificity of 92%. CONCLUSIONS: Our study suggests that endocrine secretions of Leydig and Sertoli cells are differently impacted by dialysis, KT and immunosuppressive regimen raising new issues to explore.

CONTEXTE: Les hommes insuffisants rénaux chroniques présentent souvent des signes cliniques et biologiques d'une atteinte de l'axe hypothalamo-hypophyso-testiculaire. Il est bien établi que la dialyse n'améliore pas ces anomalies endocriniennes mais l'impact de la transplantation rénale reste encore controversé. Le but de l'étude est d'explorer les taux circulants des gonadotrophines, de la testostérone et de l'inhibine B durant la période de dialyse et après la greffe de rein. PATIENTS ET MÉTHODES: Cette étude longitudinale et prospective est réalisée dans un centre hospitalo-universitaire. Les taux sériques des hormones sont mesurés par immunodosage chez 53 patients (âge moyen : 37 ans) durant la période de dialyse (T0) et 6 mois après la greffe rénale (T180). Ces données sont comparées à celles de 46 donneurs de sperme fertiles (âge moyen : 37 ans). Le principal critère de jugement est la différence des taux hormonaux entre les trois groupes. Le critère secondaire est la comparaison des taux pré- et post-greffe en fonction du traitement immunosuppresseur. RESULTATS: Durant la période de dialyse, les taux moyens de LH et de FSH sont élevés alors que ceux de testostérone et inhibine B sont normaux. Après la greffe, la LH revient à la normale alors que la FSH augmente. Les taux de testostérone restent normaux mais ceux d'inhibine B augmentent significativement. Par ailleurs, l'étude montre que la combinaison tacrolimus + mycophénolate diminue clairement les taux post-greffe d'inhibine B. Elle montre également que les taux pré-greffe d'inhibine B sont indépendants de ceux de la testostérone et qu'ils peuvent prédire les taux bas d'inhibine B avec une sensibilité de 77% et une spécificité de 92%. CONCLUSIONS: L'étude suggère que les sécrétions endocrines des cellules de Leydig et de Sertoli sont impactées de manière différente par la dialyse, la transplantation rénale et le traitement immunosuppresseurs ouvrant ainsi de nouvelles perspectives d'investigation.

Impact of chemotherapy and radiotherapy for testicular germ cell tumors on spermatogenesis and sperm DNA: a multicenter prospective study from the CECOS network.

OBJECTIVE: To determine the consequences of adjuvant testicular germ cell tumor treatment (TGCT) on sperm characteristics and sperm DNA, and to evaluate the predictors of sperm recovery. DESIGN: Multicenter prospective longitudinal study of patients analyzed before treatment and after 3, 6, 12, and 24 months. SETTING: University hospitals. PATIENT(S): One hundred twenty-nine volunteer TGCT patients and a control group of 257 fertile men. INTERVENTION(S): Routine semen analyses, sperm DNA, and chromatin assessments. MAIN OUTCOME MEASURE(S): Comparisons of mean sperm characteristics before and after treatment, with sperm recovery analyzed by the Kaplan-Meier method. RESULT(S): The quantitative and qualitative sperm characteristics decreased after treatment, with lowest values at 3 and 6 months and with variations according to treatment type. The mean total sperm count recovered to pretreatment values at 12 months after treatment after two or fewer bleomycin, etoposide, and cisplatin (BEP) cycles, but not after radiotherapy or more than two BEP cycles. Only the treatment modalities and pretreatment sperm production were related to recovery of the World Health Organization reference sperm values. An increased proportion of patients had elevated high sperm DNA stainability at 6 months after radiotherapy. CONCLUSION(S): Adjuvant treatments for testicular germ cell tumor have drastic effects on spermatogenesis and sperm chromatin quality. These new data on both the recovery period according to treatment modalities and the post-treatment chromatin status of sperm are useful tools for counseling patients wishing to conceive.

Treatment discontinuation in couples consulting for male infertility after failing to conceive.

OBJECTIVE: To evaluate rates and reasons for treatment discontinuation in couples with male factor infertility and who failed to conceive. DESIGN: Retrospective study. SETTING: Male Sterility Center, University Hospital. PATIENT(S): A total of 407 couples consulting for male factor infertility and who discontinued treatment without conceiving. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Treatment, reasons for dropout, and reproductive outcomes after discontinuation. RESULT(S): Of the 407 patients, 218 (54%) had had fertility treatment (medical or surgical), and 189 (46%) underwent assisted reproductive techniques (ART) (intrauterine insemination [IUI], in vitro fertilization [IVF], or intracytoplasmic sperm injection [ICSI]). The main reasons for dropout were painfulness of treatment (15% for patients with non-ART treatment vs. 32% for patients who had undergone ART), its ineffectiveness (12% vs. 26%), and separation of the couple (18% vs. 7%). Of the 407 patients, 27% consulted in another fertility center, 8% succeeded in having a child by ART with male partner sperm, 1% by ART with donor sperm, and 11% through adoption. CONCLUSION(S): About half of the couples consulting for male factor infertility discontinued fertility treatment, and of those who discontinued only a fifth finally succeeded in having a child. Although support is available to couples during fertility care, ART is a physical and psychological burden.

Cumulative parenthood rates in 1735 couples: impact of male factor infertility.

BACKGROUND: Most studies assessing the outcome of assisted reproductive technologies (ARTs) have reported live birth rates in couples by taking mainly the female factor into account. However, infertility is a couple's concern, and the majority of publications do not take into consideration the true impact of male infertility on having the desired number of children. METHODS: We carried out a follow-up study to evaluate the probability of having a child during treatments at the Toulouse Male Sterility Centre and after discontinuation from 2000 through 2008. Couples were followed for at least 4 years until discontinuation of treatment or delivery of a live infant. RESULTS: We were able to contact 65% of the 1735 male partners by telephone. Of the 1131 respondents, 56% had become parents (60% if adoption is included), 28% after ART, 16% by natural pregnancy, 8% after non-ART treatment and 4% after ART in another centre. The cumulative rates of success reached 64% [95% confidence interval (CI), 60-67] for men ≤35 years and women ≤35 years after 9 years, and 31% (95% CI, 24-39) in older patients. With optimistic analysis, which assumes that patients for whom no information was available have the same chance of success in having a child as those whose reproductive outcome was known, the cumulative rate of success was 48% (95% CI, 45-50) in the 1735 couples. CONCLUSIONS: More than half of couples consulting for male infertility succeeded in having a child. Male age over 35 years old appears as a key risk factor as well as the woman's age, and these findings should encourage couples to attempt parenthood earlier.

Decrease of mitochondrial DNA level in sperm from patients infected with human immunodeficiency virus-1 linked to nucleoside analogue reverse transcriptase inhibitors.

OBJECTIVE: To study sperm parameters in patients infected with human immunodeficiency virus (HIV)-1 and to analyze mitochondrial DNA (mtDNA) in sperm according to the HIV treatment. DESIGN: Observational study. SETTING: University-affiliated teaching hospital. PATIENT(S): Thirty-two patients infected with HIV-1 and 31 noninfected healthy men provided semen samples. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): After DNA extraction, mtDNA level was assessed using real-time polymerase chain reaction (PCR) (LightCycler) in whole semen and in selected spermatozoa from 90% density centrifugation gradients. For each sample, mtDNA and ß-globin gene sequences were amplified and PCR products were quantified. The mtDNA-to-ß-globin ratio expressed the number of mtDNA copies per cell. RESULT(S): Compared with the control group, several sperm parameters were altered in patients with HIV. The number of mtDNA copies per cell in whole semen was increased in HIV-infected patients (6.3±6.3 vs. 3.5±3.2). However, there was no statistically significant difference in mtDNA copy number in the spermatozoa obtained after density gradient centrifugation. The number of nucleoside analogue reverse transcriptase inhibitors (NRTI) taken by patients during treatment significantly influenced the mtDNA level in sperm (1 NRTI 7.6±8.1, 2 NRTIs 7.0±5.1, 3 NRTIs 3.2±2.1). CONCLUSION(S): Using a specific method to measure sperm mtDNA, we demonstrated a decrease of mtDNA copies in spermatozoa after use of NRTIs with known mitochondrial toxicity.

Ribavirin and pegylated interferon treatment for hepatitis C was associated not only with semen alterations but also with sperm deoxyribonucleic acid fragmentation in humans.

OBJECTIVE: To report the effects of ribavirin plus pegylated interferon on semen parameters and sperm DNA integrity in a man given treatment for hepatitis C. DESIGN: Case report. SETTING: University-affiliated teaching hospital. INTERVENTION(S): None. PATIENT(S): A 37-year-old man given treatment with ribavirin and pegylated interferon for hepatitis C. MAIN OUTCOME MEASURE(S): Semen parameters (sperm count, motility, round cells), sperm protamination state measured by sperm chromatin structure assay, and sperm DNA fragmentation measured by terminal uridine nick-end labeling assay. RESULT(S): The percentage of progressive spermatozoa and the number of motile sperm per ejaculate decreased during treatment. The round cell/spermatozoa ratio, which reflects spermatogenic abnormality, increased from 2.6% +/- 1.4% to 23.6% +/- 13.0% during treatment and returned to baseline value 4 months later. Moreover, the sperm DNA fragmentation index (as measured by sperm chromatin structure assay) increased very markedly during treatment (from 14.5% before to 69.2% at 7 months of treatment) and remained elevated 8 months later. CONCLUSION(S): This study reports for the first time not only quantitative but also qualitative alterations of spermatogenesis with DNA packaging abnormalities and emphasizes the need for prospective clinical studies. While the results of other studies are awaited, the alterations that persisted 8 months after treatment argue for a longer contraception period after treatment discontinuation in men.

Doubling of testicular cancer incidence rate over the last 20 years in southern France.

In recent decades, testicular cancer incidence has considerably increased in a majority of industrialized countries. In France, short reports suggested that the testicular cancer incidence rate has also risen, especially in north-eastern regions. In Europe, geographical variation of incidence rates has been observed in Baltic countries and a clear birth cohort effect has been revealed. This study aimed to assess temporal trends in testicular cancer incidence in southern France. We examined incidence rates over a 20-year time period in a series of 506 consecutive cases of testicular cancer recorded from 1980 to 1999 in the Midi-Pyrenees region of France. Age, calendar period, and birth cohort effects were examined simultaneously using Poisson regression models. Our analysis found a significant rise in the overall incidence rate of testicular germ cell tumors from 1.27 to 3.04 per 100,000 between 1980-1984 and 1995-1999, an annual increase of 5.70%. These results, the first obtained in a large series in southern Europe, show a twofold increase in incidence rate of testicular cancer in the Midi-Pyrenees region, which is very similar to that observed in all European countries, more or less doubling in the last 20 years. Interestingly, this major jump and the apparent testicular cancer gradient between northern and southern Europe suggest considerable geographical heterogeneity in incidence, but low geographical variation in temporal trends.

Sperm deoxyribonucleic acid fragmentation as assessed by the sperm chromatin dispersion test in assisted reproductive technology programs: results of a large prospective multicenter study.

OBJECTIVE: To assess the value of sperm DNA fragmentation, measured by the sperm chromatin dispersion (SCD) test, in predicting fertilization rate, embryo quality, and pregnancy outcome. DESIGN: Prospective study. SETTING: Four French infertility centers, from January to August 2005. PATIENT(S): Six hundred twenty-two couples participating in their first IVF or ICSI program. INTERVENTION(S): Analysis of DNA fragmentation by the sperm chromatin dispersion test in sperm samples used for IVF or ICSI. MAIN OUTCOME MEASURE(S): Correlations and associations between sperm parameters, sperm DNA integrity, and pregnancy outcomes. RESULT(S): A statistically significant correlation was observed between sperm DNA fragmentation rate and the following sperm characteristics: sperm motility, morphology, and concentration. We found a statistically significant relationship between sperm DNA fragmentation rate and fertilization rate, and we were able to suggest a threshold sperm DNA fragmentation rate of 18%, above which fragmentation rate was predictive of fertilization rate. Regarding embryo quality, we observed a relationship between sperm DNA fragmentation and embryo quality. No significant relationship was found between sperm DNA fragmentation rate and clinical pregnancies or births. CONCLUSION(S): The results of this study confirm the utility of the sperm chromatin dispersion test for assessment of DNA fragmentation.

Polyorchidism: presentation of 2 cases, review of the literature and a new management strategy.

OBJECTIVES: To present 2 cases with polyorchidism, to review the literature about polyorchidism, and to propose a rational therapeutic algorithm. METHODS: In our institution, we encountered 2 patients with polyorchidism: The 1st patient had bilateral double testis with testicular torsion, and the 2nd patient presented with an indolent scrotal mass. We also performed a literature search (PubMed) for other reports of polyorchidism. RESULTS: Polyorchidism is a rare congenital anomaly that is not well known by most urologists. So far, as illustrated by our 2 cases, the management of polyorchidism is rarely conservative, and usually the supernumerary testis is removed without any evidence supporting this attitude. Rare cases are complicated by torsion (case 1), cryptorchidism, or testicular neoplasm. In case of torsion, the conservative approach depends on the viability of the twisted testis. In case of cryptorchidism, notably in children or young adults, conservative management should be proposed, if technically feasible. In case of signs of malignancy, orchiectomy must be performed. CONCLUSIONS: Conservative treatment is advised in all uncomplicated cases. Complicated cases need a careful management, but several situations can be managed conservatively. Based on the literature, we propose a simple, rational therapeutic algorithm.