Jean-Marie Grillo

Sperm cryopreservation before cancer treatment: a 15-year monocentric experience

Sperm banking is an important procedure to preserve fertility before cancer therapy. The aim of this study was to comprehensively analyse cryopreservation activity retrospectively for 1080 patients referred to the sperm bank for sperm cryopreservation before cancer treatment. This study included 1007 patients diagnosed with testicular cancer (TC) (41.7%), lymphoma (26%), other haematological cancers (9.4%) or other types of cancer (22.8%); of these, 29 patients did not produce any semen sample and cryopreservation was impossible for 67 patients. Semen characteristics before treatment were within normal ranges, except moderate asthenospermia. Sperm concentration was significantly lower in TC than in non-TC. Straws from 57 patients (6.3%) were used in assisted reproductive technologies, which led to a 46.8% cumulative birth rate. Straws were destroyed for 170 patients (18.7%) and 140 patients performed semen analyses after cancer therapy. After an average delay of 22.5 months after the end of therapy, 43 patients (30.7%) exhibited azoospermia. This study of a large population of cancer patients revealed a high level of successful sperm storage. Utilization of cryopreserved spermatozoa led to good chances of fatherhood. Nevertheless, sperm banks should be aware of the low rates of straw use and straw destruction by cancer patients.

Occupational Exposures Obtained by Questionnaire in Clinical Practice and Their Association With Semen Quality

In industrial countries, evidence suggests that semen quality has been steadily decreasing over the past 5 decades. We employed a short questionnaire to examine the association between self-reported physical or chemical occupational exposures and semen quality. The study included 402 men consulting for couple infertility (314 with oligospermia, asthenospermia, or teratospermia and 88 with normal semen; World Health Organization criteria). Exposure effects on global sperm quality and total sperm count, sperm motility, and sperm morphology were investigated. We found significant associations between semen impairment and occupational risk factors such as exposure to heavy metals (adjusted odds ratio [OR] = 5.4; 95% confidence interval [CI], 1.6-18.1), solvents (OR = 2.5; 95% CI, 1.4-4.4), fumes (OR = 1.9; 95% CI, 1.1-3.4), and polycyclic aromatic hydrocarbons (OR = 1.9; 95% CI, 1.1-3.5). Exposure to pesticides or cement was nearly significant (OR = 3.6; 95% CI, 0.8-15.8, and OR = 2.5; 95% CI, 0.95-6.5, respectively). Physical risk factors were associated with some sperm anomalies, such as mechanical vibrations with oligospermia and teratospermia as well as excess heat and extended sitting periods with impaired motility. Exposure to ionizing radiation and electromagnetic fields was not associated with semen impairment; these results, however, may be skewed, because very few subjects reported such exposure. Despite the small dataset, self-reported exposures were correlated with semen impairment. This approach may be recommended in routine clinical practice to seek relationships between occupational exposures to reprotoxic agents and impaired semen parameters. This knowledge would allow preventive measures in the workplace to be established and could be complemented by the use of biomarkers to better characterize exposure to chemical substances and their spermiotoxic effects.

Prognosis factors of pregnancy after intrauterine insemination with the husband's sperm: conclusions of an analysis of 2,019 cycles

OBJECTIVE To identify the prognostic factors for pregnancy after intrauterine insemination with the husbands sperm (IUI-H). DESIGN Retrospective study. SETTING A single university medical center. PATIENT(S) 851 couples, for 2,019 IUI-H cycles. INTERVENTION(S) After controlled ovarian stimulation, IUI-H performed 36 hours after ovulation triggering or 24 hours after a spontaneous luteinizing hormone (LH) surge. MAIN OUTCOME MEASURE(S) Clinical pregnancy rate per cycle (PR) and delivery rate per cycle (DR). RESULT(S) The overall PR was 14.8% and DR 10.8%. Higher PR and DR were observed for patients presenting with ovulation disorders (particularly polycystic ovary syndrome) or with male infertility. Secondary infertility in the woman appeared to be a positive prognostic factor as did a basal follicle-stimulating hormone (FSH) level ≤ 7 IU/L and ovulation triggering over spontaneous LH rise. The other parameters influencing the results were the womens age, the number of mature follicles obtained (≥ 2), the endometrial thickness (10-11 mm), and the number of progressive motile spermatozoa inseminated (>1 million). CONCLUSION(S) In women aged ≤ 38 years, IUI-H should be considered as an option, particularly in cases of female infertility from ovulation disorders, in cases of a normal ovarian reserve, in cases of secondary infertility, or when ≥ 1 million progressive sperm are inseminated. Bifollicular stimulation is required. In other cases, in vitro fertilization should be discussed as the first-line treatment.

Tobacco consumption and benzo(a)pyrene-diol-epoxide–DNA adducts in spermatozoa: in smokers, swim-up procedure selects spermatozoa with decreased DNA damage

OBJECTIVE To analyze the distribution of benzo(a)pyrene-diol-epoxide (BPDE)-DNA adducts in spermatozoa selected and nonselected by a swim-up procedure with relation to smoking habits. DESIGN Comparative study. SETTING Public university and public university hospital. PATIENT(S) Seventy-nine men (37 smokers and 42 nonsmokers) who visited an infertility clinic for diagnostic. INTERVENTION(S) Tobacco and environmental exposure assessment, semen sample analysis, swim-up procedure, BPDE-DNA adduct immunolabeling. MAIN OUTCOME MEASURE(S) BPDE-DNA adduct quantification in selected (SEL-SPZ) and nonselected (NONSEL-SPZ) spermatozoa. Data were normalized by using a normalized fluorescence value (NFV). RESULT(S) The mean NFV (±SD) in SEL-SPZ was significantly higher in smokers than in nonsmokers (18.9±11.5 vs. 10.5±10.4, respectively). Within smokers, a paired analysis (SEL-SPZ and NONSEL-SPZ) showed that NFV was significantly lower in SEL-SPZ than in NONSEL-SPZ (20.0±11.3 vs. 31.5±16.0, respectively). Conversely, within nonsmokers, the mean NFV was higher in SEL-SPZ than in NONSEL-SPZ (10.3±10.6 vs 4.3±7.1, respectively). CONCLUSION(S) Tobacco consumption is associated with BPDE-DNA adducts in spermatozoa. In smokers, semen processing by swim-up recovers potentially fertilizing spermatozoa that show a significantly lower amount of BPDE-DNA adducts compared with NONSEL-SPZ. Further study is needed to improve the spermatozoa selection in smoking patients requiring assisted reproductive technologies.

Nuclear envelope remodelling during human spermiogenesis involves somatic B-type lamins and a spermatid specific B3 lamin isoform

The nuclear lamina (NL) is a filamentous protein meshwork, composed essentially of lamins, situated between the inner nuclear membrane and the chromatin. There is mounting evidence that the NL plays a role in spermatid differentiation during spermiogenesis. The mouse spermatid NL is composed of the ubiquitous lamin B1 and the spermatid-specific lamin B3, an N-terminally truncated isoform of lamin B2. However, nothing is known about the NL in human spermatids. We therefore investigated the expression pattern and localization of A-type lamins (A, C and C2) and B-type lamins (B1, B2 and B3) during human spermiogenesis. Here, we show that a lamin B3 transcript is present in human spermatids and that B-type lamins are the only lamins detectable in human spermatids. We determine that, as shown for their mouse counterparts, human lamin B3, but not lamin B2, induces strong nuclear deformation, when ectopically expressed in HeLa cells. Co-immunofluorescence revealed that, in human spermatids, B-type lamins are present at the nuclear periphery, except in the region covered by the acrosome, and that as the spermatid matures the B-type lamins recede towards the posterior pole. Only lamin B1 remains detectable on 33-47% of ejaculated spermatozoa. On spermatozoa selected for normal head density, however, this fell to <6%, suggesting that loss of the NL signal may be linked to complete sperm nucleus compaction. The similarities revealed between lamin expression during human and rodent spermiogenesis, strengthen evidence that the NL and lamin B3 have conserved functions during the intense remodelling of the mammalian spermatid nucleus.

Should sperm be cryopreserved after spinal cord injury

In spinal cord injured (SCI) patients, three main factors may cause infertility: erectile dysfunction, ejaculatory dysfunction and impaired semen quality. This letter aims to discuss how we can manage SCI patients’ fertility in accordance with patient-centred care. For such SCI patients aged 20 to 40, having children represents hope for the future. Furthermore, it is a way to rebuild a life after the spinal injury and must be seen as an important part of the rehabilitation program. We suggest that sperm cryopreservation may contribute to patient-centred care management of SCI patients’ fertility, although there is no scientific evidence that cryopreservation will improve fertility outcome after SCI. Indeed, sperm cryopreservation is an affordable and simple technique in specialised centres with trained staff. Here, a protocol to manage SCI patients’ fertility is discussed: we propose PVS for sperm banking to all SCI patients after the phase of spinal shock during the rehabilitation program. If live sperm are retrieved, they are frozen and stored; however, if no live sperm are retrieved, electroejaculation and/or surgical sperm extraction are proposed only for patients who desire biological fatherhood. Prospective studies on the evolution of semen parameters, ejaculatory dysfunction, post-infectious obstructions and spermatogenesis impairment in chronic SCI patients are urgently needed to provide robust data for the evidence-based management of SCI patients’ fertility. Even if use rates are expected to be low, sperm banking may be a simple and affordable preventative measure for selected male SCI patients.RésuméChez les patients blessés médullaires (BM), trois facteurs principaux peuvent provoquer une infertilité : la dysfonction érectile, la dyséjaculation et l’altération des paramètres spermatiques. Cette lettre a pour but de discuter comment gérer la fertilité des BM en accord avec le principe de soins axés sur le patient. Pour ces patients principalement âgés de 20 à 40 ans, la paternité représente un espoir pour le futur et un moyen de se reconstruire après la lésion médullaire ; cet aspect doit être considéré comme une partie importante du programme de réhabilitation. Nous suggérons que la cryoconservation de sperme (CS) chez le BM pourrait contribuer au principe de soins axés sur le patient, bien qu’il n’y ait pas de preuve scientifique que la CS améliore les résultats en assistance médicale à la procréation. En effet, la CS est une technique abordable et simple dans les centres spécialisés au personnel entraîné. Un protocole de prise en charge de la fertilité du BM est discuté : nous proposons la stimulation vibratoire pénienne à tous les patients BM après la phase de choc spinal, au cours du séjour en rééducation fonctionnelle. Si des spermatozoïdes sont obtenus dans l’éjaculat, ils sont cryoconservés ; dans le cas contraire, nous proposons une électroéjaculation et/ou un prélèvement chirurgical de spermatozoïdes seulement aux patients présentant un désir parental actuel. Afin d’assurer une gestion de la fertilité des BM basée sur des preuves scientifiques solides, des études prospectives de l’évolution à long terme des paramètres spermatiques, de la capacité à éjaculer, des obstructions post-infectieuses du tractus séminal et des altérations de la spermatogenèse chez les patients BM chroniques sont urgemment nécessaires. Dans l’attente de ces données, même si les taux d’utilisation attendus des paillettes sont faibles, la CS pourrait être une mesure préventive simple et abordable pour les patients BM.

Case report of apoptosis in testis of four AZFc-deleted patients: increased DNA fragmentation during meiosis, but decreased apoptotic markers in post-meiotic germ cells

AZFc deletions of the Y chromosome are the major known genetic cause of spermatogenetic failure. Meiotic studies have shown a prevalence of synaptonemal complex fragmentation and an excess of early-stage sperm cells, suggesting that the maturation block could involve apoptosis. We present a prospective and observational study of apoptotic markers in the sperm of four AZFc-deleted patients and two non-obstructive azoospermic controls without an AZFc deletion. Polycaspases assays and terminal deoxynucleotidyl transferase dUDP nick-end labelling (TUNEL) assays were combined to evaluate the incidence of apoptosis in pre-meiotic, meiotic and post-meiotic germs cells identified, respectively, using anti-melanoma-associated antigen A4 (MAGE-A4), anti-synaptonemal complex protein 3 (SCP3) and anti-sperm acrosome membrane-associated protein 1 (SPACA1) antibodies. We detected apoptosis at all stages of AZFc-deletion spermatogenesis. Using the caspase assay, the incidence of positive cells was found to be heterogeneous for pre-meiotic (from 4.8 to 84.5%) and meiotic stages (from 7.9 to 57.6%), while for post-meiotic cells, the mean incidence was 6% in AZFc-deleted patients compared with 26.5% in controls (P < 0.05). Using the TUNEL assay, the mean percentage with DNA fragmentation for meiotic cells was 54.0% in AZFc-deleted patients compared with 20.3% in controls (P < 0.05), while the percentage of TUNEL-positive post-meiotic cells ranged from 5.3 to 44.7%. Spermatocyte loss in AZFc-deleted patients occurs via the apoptotic pathway. In post-meiotic cells, the lower incidence of apoptosis in testis from three of the four AZFc-deleted patients, compared with controls, is consistent with AZFc deletions having little negative impact on sperm quality.

Association entre anomalies spermatiques et environnement professionnel chez les hommes consultant pour infertilité de couple

Alteration of sperm parameters related to occupational exposures is the subject of several studies, often on a case-control approach. The study populations usually comprise men consulting in infertility clinics for couple infertility. The objective of this review is to identify, from these case-control studies, the main occupational factors that may be associated with altered sperm parameters. We selected 13 articles in the PubMed database. Participation in these studies varied from 61 to 2619 subjects, with great methodological heterogeneity, particularly in the characterization of exposure. The main occupations that appear significantly associated with a risk of altered sperm parameters are workmen, painters, farmers, welders, plumbers and technicians. When analysis focuses on occupational exposures, a significant result is reported for solvents, heavy metals, heat, vibrations and non-ionizing radiation. None of the selected studies has found a link with exposure to pesticides.