BACKGROUND:No study has yet explored telomere length or interaction in sperm nuclei of testicular cancer (TC) patients exposed to chemotherapy or radiotherapy. However, sperm telomere dynamics have emerged as a potential marker in male infertility. OBJECTIVES:We aimed to investigate in a pilot and exploratory study whether TC and its adjuvant treatments alter sperm telomeres and DNA integrity during a 2-year follow-up after treatment. MATERIALS AND METHODS:This ancillary study is part of the multicentric prospective and longitudinal French GAMATOX study. Twenty-nine TC patients treated with orchiectomy and either adjuvant chemotherapy (n = 14) or radiotherapy (n = 15) were included, alongside ten fertile donors. Semen samples were collected before orchiectomy and adjuvant therapy, and at 3, 6, 12, and 24 months after treatment. Sperm telomere length (STL) and telomere interactions were assessed by quantitative fluorescent in situ hybridisation (FISH). Sperm DNA fragmentation (SDF) was measured using the TUNEL assay, aneuploidy by FISH, and chromatin condensation defects by aniline blue staining. RESULTS:Before adjuvant therapy, patients presented a higher number of sperm telomere signals compared with controls (p = 0.004), persisting 2 years after treatment (p = 0.001). STL was not affected by tumour histology. Extremely short telomeres and the highest number of telomere signals per nucleus were more frequently observed after chemotherapy. SDF and chromatin condensation defects were transient and paralleled the recovery of conventional semen parameters, regardless of treatment type. A moderate increase in sperm aneuploidy was observed 6 months after chemotherapy (p = 0.03). CONCLUSION:TC patients presented impaired sperm telomere interactions before and after adjuvant therapy, as reflected by a persistently elevated number of telomere signals per nucleus, and a higher prevalence of extremely short sperm telomeres, especially after chemotherapy. While relative STL remained comparable to controls, these findings raise questions about the impact of telomere architecture on germ cell tumour biology and post-treatment fertility.

OBJECTIVES: Etiological factors of testicular germ cell tumors (TGCT) remain largely unknown, but a causal role of occupational exposures to solvents has been suggested. Previous studies analyzing these exposures reported discordant results, potentially related to exposure assessment methods. The aim of this study was to investigate the role of occupational exposure to solvents on the risk of developing TGCT among young men. METHODS: This study examined occupational exposures to solvents and TGCT risk based on the lifetime work histories of 454 cases and 670 controls, aged 18–45 years, of the French national TESTIS case–control study. Solvent exposure was estimated using: (i) exposure assignment by job-exposure matrix (JEM) and (ii) JEM combined with self-reported exposure data from specific questionnaires (SQ) and expert assessment (EA). Odds ratios (OR) and 95% confidence intervals (CI) were estimated using conditional logistic regression models. RESULTS: Both approaches (JEM and JEM+SQ+EA) showed a consistent association between TGCT and trichloroethylene exposure (exposed versus not exposed; JEM=OR 1.80 [95% confidence interval (CI) 1.12–2.90] and JEM+SQ+EA= OR 2.59 (95% CI 1.42–4.72). Both approaches also observed positive associations with ketone esters and fuels & petroleum-based solvents. CONCLUSION: The results suggest that some organic solvents might be involved in the pathogenesis of TGCT among occupationally exposed men. The combined use of JEM+SQ+EA seemed to limit misclassification by considering individual exposure variability and is, therefore, an appealing approach to assess occupational exposures in epidemiological studies.

BackgroundSevere acute respiratory syndrome coronavirus 2, the virus responsible for coronavirus disease 2019, affects multiple organs. The virus enters cells through angiotensin-converting enzyme-2 and host factors present in genital organs, leading to concern over virus shedding in semen and reproductive function.ObjectivesTo investigate severe acute respiratory syndrome coronavirus 2 in semen from patients with a mild infection, identify the seminal infected cells, and explore the effect of the infection on sex hormones and semen parameters.Materials and methodsProspective study of 54 men with mild severe acute respiratory syndrome coronavirus 2 infection. Semen was collected at 7, 15, 30, 60, 90, 180, and 365 days after symptom onset, and severe acute respiratory syndrome coronavirus 2 RNA was measured in serum, saliva, urine, and semen. The presence of infectious severe acute respiratory syndrome coronavirus 2 in semen was assessed using Vero cell culture. Infected semen cells were identified using immunofluorescence against severe acute respiratory syndrome coronavirus 2 nucleoprotein antigen and cell markers. Semen characteristics as well as testosterone, inhibin B, luteinizing hormone, and follicle-stimulating hormone levels were determined.Results11% of patients had at least one severe acute respiratory syndrome coronavirus 2 RNA-positive semen. One patient had viral semen shedding up to day 90 after infection onset, with replication-competent virus isolated from semen and 40% cell fraction at day 7. After sperm preparation, 90% fraction was severe acute respiratory syndrome coronavirus 2 RNA-positive at days 7 and 15. The swim-up fraction was positive only on day 7. In semen, nucleoprotein antigen was detected mainly in exfoliated epithelial cells and less frequently in Sertoli cells. Sperm count and motile sperm count were lower at day 30 than at day 7. Round cells in semen were increased during the acute phase. At days 7 and 15, sperm count and motile sperm count were lower in severe acute respiratory syndrome coronavirus 2 RNA-positive semen compared with negative semen, while semen volume and follicle-stimulating hormone levels were increased. Long-term follow-up shows no evidence of a detrimental effect on hormonal or semen characteristics.Discussion and conclusion11% of patients with mild coronavirus disease 2019 who were not hospitalized had severe acute respiratory syndrome coronavirus 2 excretions in semen, which persisted for up to 90 days in one patient. No germ cells appeared infected by the virus, but the detection of nucleoprotein antigen-positive epithelial semen cells and Sertoli cells suggests genital tract infection. Albeit infrequent, semen may contain the replication-competent virus during the acute phase with potential risk of severe acute respiratory syndrome coronavirus 2 transmissions during sexual contact and assisted reproduction procedures. The effect of mild coronavirus disease 2019 on spermatogenesis and reproductive hormones was moderate and reversible.

STUDY QUESTION:What is the impact of low- or moderate-risk gonadotoxic chemotherapy received prior to testicular tissue freezing (TTF), and of the cancer itself, on spermatogonia quantity in testicular tissue from (pre)pubertal boys?SUMMARY ANSWER:Vincristine, when associated with alkylating agents, has an additional adverse effect on spermatogonia quantity, while carboplatin has no individual contribution to spermatogonia quantity, in testicular tissue of (pre)pubertal boys, when compared to patients who have received non-alkylating chemotherapy.WHAT IS KNOWN ALREADY:The improved survival rates after cancer treatment necessitate the inclusion of fertility preservation procedures as part of the comprehensive care for patients, taking into consideration their age. Sperm cryopreservation is an established procedure in post-pubertal males while the TTF proposed for (pre)pubertal boys remains experimental. Several studies exploring testicular tissue of (pre)pubertal boys after TTF have examined the tubular fertility index (TFI, percentage of seminiferous tubule cross-sections containing spermatogonia) and the number of spermatogonia per seminiferous tubule cross-section (S/T). All studies have demonstrated that TFI and S/T always decrease after the introduction of chemotherapeutic agents, especially those which carry high gonadotoxic risks such as alkylating agents.STUDY DESIGN, SIZE, DURATION:Testicular tissue samples from 79 (pre)pubertal boys diagnosed with cancer (from 6 months to 16 years of age) were cryopreserved between May 2009 and June 2014. Their medical diagnoses and previous chemotherapy exposures were recorded. We examined histological sections of (pre)pubertal testicular tissue to elucidate whether the chemotherapy or the primary diagnosis affects mainly TFI and S/T.PARTICIPANTS/MATERIALS, SETTING, METHODS:(Pre)pubertal boys with cancer diagnosis who had been offered TTF prior to conditioning treatment for hematopoietic stem cell transplantation were included in the study. All the patients had previously received chemotherapy with low- or moderate-risk for future fertility. We have selected patients for whom the information on the chemotherapy received was complete. The quantity of spermatogonia and quality of testicular tissue were assessed by both morphological and immunohistochemical analyses.MAIN RESULTS AND THE ROLE OF CHANCE:A significant reduction in the number of spermatogonia was observed in boys treated with alkylating agents. The mean S/T values in boys exposed to alkylating agents were significantly lower compared to boys exposed to non-alkylating agents (P = 0.018). In contrast, no difference was observed for patients treated with carboplatin as the sole administered alkylating agent compared to the group of patients exposed to non-alkylating agents. We observed an increase of S/T with age in the group of patients who did not receive any alkylating agent and a decrease of S/T with age when patients received alkylating agents included in the cyclophosphamide equivalent dose (CED) formula (r = 0.6166, P = 0.0434; r = -0.3759, P = 0.0036, respectively). The TFI and S/T decreased further in the group of patients who received vincristine in combination with alkylating agents (decrease of 22.4%, P = 0.0049 and P < 0.0001, respectively), but in this group the CED was also increased significantly (P < 0.0001). Multivariate analysis, after CED adjustment, showed the persistence of a decrease in TFI correlated with vincristine administration (P = 0.02).LIMITATIONS, REASONS FOR CAUTION:This is a descriptive study of testicular tissues obtained from (pre)pubertal boys who were at risk of infertility. The study population is quite heterogeneous, with a small number of patients in each sub-group. Our results are based on comparisons between patients receiving alkylating agents compared to patients receiving non-alkylating agents rather than chemotherapy-naive patients. The French national guidelines for fertility preservation in cancer patients recommend TTF before highly gonadotoxic treatment. Therefore, all the patients had received low- or moderate-risk gonadotoxic chemotherapy before TTF. Access to testicular tissue samples from chemotherapy-naive patients with comparable histological types of cancer was not possible. The functionality of spermatogonia and somatic cells could not be tested by transplantation or in vitro maturation due to limited sample sizes.WIDER IMPLICATIONS OF THE FINDINGS:This study summarizes the spermatogonial quantity of (pre)pubertal boys prior to TTF. We confirmed a negative correlation between the cumulative exposure to alkylating agents and spermatogonial quantity. In addition, the synergistic use of vincristine in combination with alkylating agents showed a cumulative deleterious effect on the TFI. For patients for whom fertility preservation is indicated, TTF should be proposed for chemotherapy with a predicted CED above 4000 mg/m2. However, the data obtained from vincristine and carboplatin use should be confirmed in a subsequent study including more patients.STUDY FUNDING/COMPETING INTEREST(S):This study had financial support from a French national research grant PHRC No. 2008/071/HP obtained by the French Institute of Cancer and the French Healthcare Organization. The sponsors played no role in the study. The authors declare no conflicts of interest.TRIAL REGISTRATION NUMBER:N/A.

Abstract Study question What is the impact of the cancer treatment received prior to TTF and the disease on spermatogonia quantity in testicular tissue from (pre)pubertal boys? Summary answer A decrease in spermatogonia number was observed in testicular tissue after cancer treatment when cyclophosphamide equivalente dose (CED) is above 4000 mg/m2. What is known already The improved survival rates associated with the development of sperm and testicular tissue freezing (TTF) renders difficult not to offer fertility preservation to children or adolescents before cancer. Several studies exploring cancer patients have examined the number of spermatogonia per seminiferous tubular cross-section (S/T) or tubular fertility index (TFI, percentage of tubular cross-sections containing spermatogonia) in testicular biopsies. All studies, demonstrated that the S/T and TFI always decreased after the introduction of chemotherapy and more specifically in case of highly gonadotoxic risk such as alkylating agents. Study design, size, duration Testicular tissue samples from 79 (pre)pubertal boys diagnosed with cancer (ranging from 6 month to 16 years of age) were cryopreserved between May 2009 and June 2014. Medical diagnosis and previous chemotherapy exposure were recorded. We examined histological sections of (pre)pubertal testicular tissue to elucidate whether chemotherapy, doses or primary diagnosis affects the quality of testicular tissue. Participants/materials, setting, methods (Pre)pubertal boys with cancer diagnosis who benefitted from TTF prior to conditioning treatment for hematopoietic stem cell transplantation. All the patients included had previously received chemotherapy with moderate risk for future fertility. We have selected patients for whom data on chemotherapy received were complete. The quantity of spermatogonia and quality of testicular tissue were assessed by both morphological and immunohistochemical analysis. Main results and the role of chance The main finding was a significant reduction in spermatogonial cell counts in boys treated with alkylating agents. The mean S/T values in boys exposed to alkylating agents was significantly lower than in a group exposed to non-alkylating agents (p = 0.018). In contrast, no difference was observed for patients treated with carboplatin as the only alkylating agent compared to the group of patients exposed to non-alkylating agents. We observed an increase of S/T with age in the group of patients who did not receive alkylating agents and a decrease of S/T with age when patients received alkylating agents included in the CED formula (r = 0.6166, p = 0.0434; r= -0.3759, p = 0.0036, respectively). The TFI and S/T were decreased in the group of patients who received vincristine (p = 0.0049; p < 0.0001, respectively), but the CED was also significantly increased (p < 0.0001). Multivariate analysis, adjusted for CED, showed the persistence of a decrease in TFI correlated with vincristine administration (-0.5 [-0.96; 0.09], p = 0.02). Limitations, reasons for caution This is a descriptive study of testicular tissues obtained from patients who were at risk of infertility. Spermatognia functionanlity could not be tested by transplantation due limited sample size. Wider implications of the findings This study summarizes spermatogonia quantity and quality of testicular tissue of (pre)pubertal boys after potentially sterilizing treatments. We confirmed a negative correlation between the cumulative exposure to alkylating agents and the spermatogonial quantity. For patients in whom fertility preservation is indicated, TTF should be performed before initiation of alkylating agents. Trial registration number N/A

STUDY QUESTION In a non-commercial national gamete donation programme, do the motivations and personality characteristics of candidate sperm and oocyte donors differ according to their parenthood status? SUMMARY ANSWER Moderate differences exist between non-parent and parent candidate donors in motivations for gamete donation and representations as well as in personality characteristics. WHAT IS KNOWN ALREADY Several studies have analysed the motivations and experiences of oocyte or sperm donors, but mainly in countries where gamete donation is a commercial transaction, and very few studies have reported results of personality traits using personality inventory tests. No study has specifically investigated the motivations and personality characteristics of candidate gamete donors according to parenthood status. STUDY DESIGN, SIZE, DURATION A prospective study was carried out including 1021 candidate donors from 21 centres (in university hospitals) of the national sperm and egg banking network in France between November 2016 and December 2018. PARTICIPANTS/MATERIALS, SETTING, METHODS In total, 1021 candidate gamete donors were included in the study. During their first visit, male (n = 488) and female candidate donors (n = 533) completed a questionnaire on sociodemographic characteristics, their motivations for donation and their representations of donation, infertility and family. Secondly, a NEO Personality Inventory (NEO-PI-R) exploring the Big Five personality traits was completed online. Results were compared between parent and non-parent candidate donors. MAIN RESULTS AND THE ROLE OF CHANCE Altruistic values were the principal motive for donation irrespective of parenthood status. Reassurance about their fertility or preservation of sperm for future use was more often reported in non-parent than in parent candidate donors. With regard to representation of gamete donation or of the family, independently of their parenthood status, candidate donors more frequently selected social rather than biological representations. Mean personality characteristics were in the normal range. Non-parent candidate donors had higher scores on openness and depression than parents, while parent candidate donors appeared more social than non-parents. LIMITATIONS, REASONS FOR CAUTION The personality characteristics inventory was not completed by all candidate donors included in the study. However, family status did not differ between the two groups (NEO-PI-R completed (n = 525) or not), while the group who completed the NEO-PI-R had a higher educational level. This national study was performed in a country where gamete donation is subject to strict legislation. WIDER IMPLICATIONS OF THE FINDINGS In a global context where reproductive medicine is commercialized and gamete donor resources are limited, this study found that altruism and social representations of gamete donation and family are the main motivations for gamete donation in a country which prohibits financial incentive. These findings are relevant for health policy and for gamete donation information campaigns. STUDY FUNDING/COMPETING INTEREST(S) Grant from the Agence de la Biomedecine, France. The authors have nothing to disclose related to this study.

Testicular tissue freezing is proposed for fertility preservation to (pre)pubertal boys with cancer before highly gonadotoxic treatment. Studies accurately comparing human (pre)pubertal testicular tissue quality before freezing and after thawing are exceptional. No study has reported this approach in a systematic manner and routine care.

Background Men with congenital unilateral absence of vas deferens were reported to be mainly azoospermic, with both unilateral renal absence and mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) but some have neither. Objectives To assess whether in infertile couples the male partners with congenital unilateral absence of vas deferens are mainly azoospermic men. Material and Methods Retrospective study in a unique university hospital; reproductive, clinical, CFTR analysis and seminal data of male partners of infertile couples (from 1998 to 2018) were analysed. Diagnosis of congenital unilateral absence of vas deferens was based on transrectal ultrasounds (TRUS): complete or partial absence of one vas deferens with complete contralateral vas deferens confirmed in 63 men. Distribution of sperm count in three classes: azoospermia, oligozoospermia or normozoospermia. Ultrasound determination of renal status; seminal biomarkers assays; and search for CFTR mutations. Results Among the 63 men, 39.7% displayed azoospermia, 27% oligozoospermia and 33.3% normozoospermia; 42% of the non-azoospermic men (16/38) had previously obtained a natural pregnancy. We found unilateral renal absence in 17/59 patients (29%). Among 50 men with CFTR testing, five carried an allele associated with cystic fibrosis belonging to the 29 men without renal anomalies, indicating a high allelic frequency (8.6%). The 63 patients displayed high rates of surgical histories for undescended testicles or inguinal hernia, low values of semen volume and of total seminal glycerophosphocholine. Conclusions Our results indicate that men with congenital unilateral absence of vas deferens mainly display oligozoospermia or normozoospermia and that they were previously fertile. They clearly confirm, first, that CFTR testing is recommended in congenital unilateral absence of vas deferens men and it should be mandatory for those with normal kidneys; and, second, that TRUS is needed for the diagnosis of congenital unilateral absence of vas deferens. As congenital unilateral absence of vas deferens may be present whatever the sperm count, biological warnings are represented by semen volume and seminal epididymal markers and clinical warnings by surgical histories of undescended testes or inguinal hernia.

Context Abnormalities in the hypothalamo-pituitary-gonadal axis have long been reported in Noonan syndrome (NS) males with only few data available in prepubertal children. Objective The aim of this study was to describe the gonadal function of NS males from childhood to adulthood. Design It is a retrospective chart review. Patients and methods A total of 37 males with a genetically confirmed diagnosis of NS were included. Clinical and genetic features, as well as serum hormone levels (LH, FSH, testosterone, anti-Müllerian hormone (AMH), and inhibin B) were analysed. Results Of the 37 patients, 16 (43%) children had entered puberty at a median age of 13.5 years (range: 11.4-15.0 years); age at pubertal onset was negatively correlated with BMI SDS (r = -0.541; P = 0.022). In pubertal boys, testosterone levels were normal suggesting a normal Leydig cell function. In contrast, NS patients had significant lower levels of AMH (mean SDS: -0.6 ± 1.1; P = 0.003) and inhibin B (mean SDS: -1.1 ± 1.2; P < 0.001) compared with the general population, suggesting a Sertoli cell dysfunction. Lower AMH and inhibin B levels were found in NS-PTPN11 patients, whereas these markers did not differ from healthy children in SOS1 patients. No difference was found between cryptorchid and non-cryptorchid patients for AMH and inhibin B levels (P = 0.43 and 0.62 respectively). Four NS-PTPN11 patients had a severe primary hypogonadism with azoospermia/cryptozoospermia. Conclusions NS males display Sertoli cell-specific primary testicular insufficiency, whereas Leydig cell function seems to be unaffected.

While reproductive technologies are increasingly used worldwide, epidemiologic, clinical and genetic data regarding infertile men with combined genital tract and renal abnormalities remain scarce, preventing adequate genetic counseling.

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.

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.

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. (C)2016 by American Society for Reproductive Medicine.

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. (C) 2016 by American Society for Reproductive Medicine.

The tropical pathogen Zika virus has been neglected for decades because it generally results in moderate or clinically asymptomatic infections in Africa and Asia. However, the 2015 outbreak in Brazil has focused attention on the impact of Zika virus on human health, as evidence accumulated indicating that it causes severe neurological injuries and adverse fetal outcomes.1Musso D Gubler DJ Zika virus.Clin Microbiol Rev. 2016; 29: 487-524Crossref PubMed Scopus (970) Google Scholar The Zika virus is a flavivirus that is primarily transmitted to human beings by mosquitoes of the genus Aedes. Other routes of transmission are via the blood, with the implied risk to blood transfusions; via semen,2Mansuy JM Dutertre M Mengelle C et al.Zika virus: high infectious viral load in semen, a new sexually transmitted pathogen?.Lancet Infect Dis. 2016; 16: 405Summary Full Text Full Text PDF PubMed Scopus (292) Google Scholar with the risk of sexual transmission;3D'Ortenzio E Matheron S de Lamballerie X et al.Evidence of sexual transmission of Zika virus.N Engl J Med. 2016; 374: 2195-2198Crossref PubMed Scopus (546) Google Scholar and via the placenta, with the risk of transmission to fetuses. Transmission via semen and the placenta are real challenges for women of reproductive age. We report the case of a 27-year-old man who had had a mass in the popliteal area for several months, and was diagnosed with a monophasic grade 2 synovial cell sarcoma in mid-February, 2016. A combination of treatment, including aggressive chemotherapy, was planned for the beginning of April, and the patient was therefore sent to the local centre for the study and conservation of human sperm and ova. French guidelines on Zika virus infection risk during fertility preservation have been designed for patients returning from epidemic areas. They recommend that fertility preservation management should be delayed for up to 28 days after travel and that semen should be checked for Zika virus within 6 months of travel. The patient therefore underwent a systematic medical interview, which revealed that he had recently travelled to southeast Asia. Although he had not travelled within any epidemic areas, samples of blood, urine, and semen were collected on March 9, 2016, and sent to the department of virology at Toulouse University Hospital for Zika virus diagnosis. No traces of the Zika virus genome were detected in the blood or urine samples, but Zika virus RNA was detected twice in the semen. The viral load was 4800 copies per mL (RealStar Zika Virus RT-PCR kit 1·0, Altona Diagnostics, Hamburg, Germany) and the strain that was detected clustered with the Asian genotype group on a 376 nucleotide fragment from the NS5 region. The semen characteristics were in the normal range. A second medical interview found that the patient had been travelling in Thailand between Oct 18 and Nov 30, 2015, and had been bitten by mosquitos, but had not been exposed to any known risk of Zika virus transmission since his return to France. Mild symptoms (asthenia, myalgia, and conjunctivitis) compatible with Zika virus infection appeared on Dec 6, 2015, and lasted for a few days. Zika virus was detected in the semen 93 days after the onset of the Zika fever symptoms. The data from this case indicate that people returning from non-epidemic areas such as Thailand can become infected with Zika virus.4Buathong R Hermann L Thaisomboonsuk B et al.Detection of Zika virus infection in Thailand, 2012–2014.Am J Trop Med Hyg. 2015; 93: 380-383Crossref PubMed Scopus (194) Google Scholar Also, they suggest that semen can carry Zika virus produced in a currently unknown replication reservoir. Third, protracted shedding into the semen should be considered when taking measures to preserve male fertility before any aggressive surgical, chemical, or radiological treatment. Lastly, the protracted shedding into the semen should be taken into account when designing measures to protect pregnant women, women trying to get pregnant, and couples using assisted reproductive technology from the risk of Zika virus infection. The 6-month risk period after symptom onset during which patients returning from areas of active Zika virus transmission should be monitored for a Zika virus infection, as recommended by French guidelines and by the US Centers for Disease Control and Prevention,5Petersen EE Polen KN Meaney-Delman D et al.Update: interim guidance for health care providers caring for women of reproductive age with possible Zika virus exposure—United States, 2016.MMWR Morb Mortal Wkly Rep. 2016; 65: 315-322Crossref PubMed Scopus (100) Google Scholar should be extended to patients returning from non-epidemic areas. It should also be regularly revised to keep pace with the evolution of scientific knowledge about the seminal shedding of Zika virus. We declare no competing interests. Zika virus: high infectious viral load in semen, a new sexually transmitted pathogen?The recent, rapid spread of Zika virus in South America and increasing reports of cases of congenital abnormalities spatiotemporally associated with Zika virus infections led WHO to declare a Public Health Emergency of International Concern on Feb 1. WHO also recently described measures that pregnant women should take to avoid infection. Full-Text PDF Should testing of donors be restricted to active Zika virus areas?Jean Michel Mansuy and colleagues1 report the detection of Zika virus in the semen of a patient returning from a non-epidemic area, even while usual recommendations limit testing to only those returning from epidemic areas.1 However, categorisation of Zika-virus-affected countries by WHO, US Centers for Disease Control and Prevention (CDC), US Food and Drug Administration (FDA), European Centre for Disease Prevention and Control (ECDC), and others differ and are often discordant and their update cycles for recommendations for those exposed in endemic and epidemic areas vary in timeliness (appendix). Full-Text PDF

STUDY QUESTION:In which cases is freezing of ejaculated sperm indicated before ICSI? SUMMARY ANSWER:Sperm freezing should be performed only when out of two analyses at least one total sperm count in the ejaculate is lower than 10(6). WHAT IS KNOWN ALREADY:Due to variations in individual sperm parameters, in cases of severe oligozoospermia there is a risk of absence of spermatozoa on the day of ICSI, leading to cancellation of the attempt. Sperm freezing can avoid this problem but little is known of the parameters governing the decision to freeze sperm or not. STUDY DESIGN, SIZE, DURATION:This retrospective study included 247 men who underwent sperm cryopreservation to prevent the risk of azoospermia on the day of ICSI, from 2000 to 2012. PARTICIPANTS/MATERIALS, SETTING, METHODS:Receiver operating characteristic curve analysis was used to define the threshold value. The lowest total sperm count per ejaculate was studied as a predictive factor for the use of frozen sperm in a total of 593 ICSI attempts. Moreover, 2003 patients who had at least 4 semen analyses for andrological diagnosis have been studied to evaluate the reproducibility of sperm count. To evaluate the psychological impact of sperm freezing, a questionnaire was administered to 84 men who attended for sperm cryopreservation between June and December 2014. The cost of sperm freezing was analysed according to the French prices. MAIN RESULTS AND THE ROLE OF CHANCE:When at least one total sperm count was <10(5) the risk of azoospermia in at least one ICSI attempt was 52% (34/66) versus 3% (5/181) when all counts were ≥10(5) (P < 0.0001). However, the study of the reproducibility of pre-ICSI semen analyses has shown wide variations among ejaculates, and therefore sperm freezing is recommended when one analysis from at least two, showed a sperm count <10(6). Such a policy could allow a saving of about €70 000 by avoiding unnecessary sperm freezings. The psychological impact of sperm freezing was good since >70% of men had positive feelings about this technique. LIMITATIONS, REASONS FOR CAUTION:This was a fairly short-term study and preservation of future fertility was not assessed. It appeared impossible to find a threshold that would predict the risk of azoospermia with 100% accuracy. Therefore there is still a risk of absence of spermatozoa on the day of ICSI despite a good negative predictive value when no total sperm count was lower than 10(5). WIDER IMPLICATIONS OF THE FINDINGS:These data suggest that sperm freezing should be proposed when total sperm count is lower than 10(6) to avoid cancellation of the ICSI attempt due to azoospermia.