Influence of sickle cell disease and treatment with hydroxyurea on sperm parameters and fertility of human males.

BACKGROUND: Recent progress in the treatment of sickle cell disease, in particular the use of hydroxyurea, has considerably modified the prognosis of this disease. Many more patients now reach reproductive age. The objective of this study was to assess the potential impact of hydroxyurea on the semen of patients. DESIGN AND METHODS: In this retrospective multicenter study, we evaluated the sperm parameters and fertility of 44 patients and analyzed the potential impact of hydroxyurea. RESULTS: We report data from the largest series so far of semen analyses in patients with sickle cell disease: 108 samples were analyzed, of which 76 were collected before treatment. We found that at least one sperm parameter was abnormal in 91% of the patients before treatment, in agreement with published literature. All sperm parameters seemed to be affected in semen samples collected during hydroxyurea treatment, and this impairment occurred in less than 6 months, later reaching a plateau. Furthermore, after hydroxyurea cessation, while global results in 30 patients were not statistically different before and after hydroxyurea treatment, in four individuals follow-up sperm parameters did not seem to recover quickly and the total number of spermatozoa per ejaculate fell below the normal range in about half the cases. CONCLUSIONS: The observed alterations of semen parameters due to sickle cell disease seem to be exacerbated by hydroxyurea treatment. Until prospective studies reveal reassuring findings, we suggest that a pre-treatment sperm analysis be performed and sperm cryopreservation be offered to patients before hydroxyurea treatment.

In Vitro fertilization failure of normozoospermic men: search for a lack of testicular isozyme of angiotensin-converting enzyme.

BACKGROUND: Angiotensin converting enzyme (ACE) is a metalloprotease with two isoforms. The somatic isoform is a key component of the renin-angiotensin system; its main function is to hydrolyse angiotensin I into angiotensin II. The germinal or testicular isoform (tACE) located at the plasma membrane of the spermatozoa, plays a crucial role in the spermatozoa-oocyte interaction during in vivo fertilization, in rodents. Disruption of the tACE in mice has revealed that homozygous male tACE-/- sire few pups despite mating normally. Few spermatozoa from these tACE-/- mice are bound to the zona pellucida (ZP) despite normal semen parameters. Based on these findings in mice models, we hypothesized that some infertile men that have the same phenotype as the tACE-/- mice, ie normal semen parameters and a lack of sperm bind to the ZP in vitro, may have a tACE defect. METHODS: Twenty four men participated to this study. The case subjects (n = 10) had normal semen parameters according to the WHO guidelines (WHO 1999) but a total in vitro fertilization failure with absence of sperm fixation to the ZP. The control subjects (n = 14) also had normal semen parameters and a normal fertilization rate ≥65%. We investigated the tACE expression in spermatozoa by Western-Blot and performed a DNA sequencing of the tACE gene. RESULTS: Three case-subjects and one control-subject had no tACE expression. There were no statistic differences between the two groups. No mutation was detected in the tACE DNA sequence. CONCLUSIONS: Our results didn't show any involvement of tACE in human fertilization especially in ZP binding.

INTRODUCTION: L'enzyme de conversion de l'angiotensine (ACE) est une métalloprotéase qui se présente sous deux isoformes. Une isoforme somatique qui intervient dans la régulation de la pression artérielle via le système rénine-angiotensine et une isoforme testiculaire spermatique (ACEt) qui interviendrait dans la fixation du spermatozoïde aux enveloppes ovocytaires (zone pellucide et/ou oelemme). Des expériences d'invalidation du gène de l'ACEt chez la souris ont montré que les mâles homozygotes ACEt−/− présentaient une faible descendance in vivo et une fixation faible des spermatozoïdes à la zone pellucide (ZP) in vitro et ce malgré un sperme normal. Ces résultats nous ont permis d'émettre l'hypothèse que certains échecs de fécondation en fécondation in vitro classique (FIV) chez des patients à sperme normal seraient en rapport avec un déficit de l'ACEt au niveau des spermatozoïdes. MATÉRIEL ET MÉTHODES: Vingt quatre patients ont été inclus dont 10 cas avec un sperme normal et un échec total de fécondation lors d'une FIV et 14 témoins ayant les mêmes caractéristiques spermatiques mais présentant de bons taux de fécondation (≥65%). L'étude de l'expression protéique de l'ACEt a été faite par western-blot sur les culots de spermatozoïdes. Un séquençage du gène codant pour l'ACEt a été fait sur l'ADN lymphocytaire. RÉSULTATS: Trois cas et un témoin présentaient un défaut d'expression de l'ACEt sans diffèrence statistiquement significative entre ces deux groupes. En PCR, aucune mutation n'a été retrouvée après séquençage du gène de l'ACEt. CONCLUSIONS: Nos résultats n'ont pas objectivé une implication de l'ACEt dans l'interaction entre ovocyte et spermatozoïde et notamment dans la fixation à la ZP.

Successful pregnancy by insemination of spermatozoa in a woman with a human seminal plasma allergy: should in vitro fertilization be considered first?

OBJECTIVE: To raise the possibility that pregnancy can be obtained by assisted reproductive techniques in patients with human seminal plasma allergy. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A woman consulted for a 3-year primary infertility. She reported lack of intercourse because of a seminal plasma allergy. INTERVENTION(S): One intrauterine insemination associated with antihistamine treatment was performed with carefully washed spermatozoa. RESULT(S): Immediately after the insemination, the patient had an allergic reaction treated by steroids. Fortunately, this single attempt led to a successful pregnancy, and the patient gave birth to a healthy girl. CONCLUSION(S): Pregnancies can be obtained in patients with seminal plasma hypersensitivity by means of intrauterine insemination. However, serious complications may occur after performing intrauterine insemination, which is not a totally reliable method to prevent an allergic reaction. Thus a multidisciplinary team should follow such patients carefully and watch for the potential risks and side effects. In vitro fertilization may represent the more cautious option.

Learning curve of vitrification assessed by cumulative summation test for learning curve (LC-CUSUM).

We used the cumulative summation test for learning curve (LC-CUSUM), a specifically designed statistical tool, to evaluate the first 50 procedures performed by a trainee in vitrification and to provide a usable model for monitoring the learning process of this technique. Given the lack of models to evaluate IVF technologies, the CUSUM methodology could prove useful for quality control in laboratories.