Tumor necrosis factor-alpha -308 polymorphism in infertile men with altered sperm production or motility.

BACKGROUND: One of the most well-documented cytokines suspected as a hazard to male fertility is tumor necrosis factor-alpha (TNFalpha). Genetic factors such as single-nucleotide polymorphisms (SNPs) in the TNF gene cluster impact TNFalpha levels. Our objective was to establish the potential involvement of -308 TNF SNP in male infertility risk. METHODS: In 684 infertile male patients undergoing an intracytoplasmic sperm injection procedure, we used allele-specific polymerase chain reaction (PCR) and PCR-RFLP to investigate the distribution of the guanine (G)-to-adenosine (A) substitution at position -308 in the promoter region of the TNFalpha gene. RESULTS: An increased frequency of the -308 TNFalpha A allele was found in patients with low sperm count of testicular origin [P = 0.002; odds ratio (OR) = 2.93] or with normal production count but altered sperm motility (P = 0.003; OR = 2.32), compared with a patient group with normal sperm count and quality (morphology and motility). In patients with low sperm count exhibiting TNFalpha A allele, compared with those with G allele, an alteration in hormonal balance was observed with increased inhibin B levels and subsequent reduced FSH plasma levels, leading to an FSH/inhibin B ratio roughly half as high (from 0.07 +/- 0.01 in TNFA versus 0.13 +/- 0.02 in TNFG allele groups, P < 0.0001). CONCLUSION: As the -308 TNFalpha A allele has been associated with an increased expression/production of TNFalpha, the potential use of therapies based on inhibition of TNFalpha activities could represent possible therapeutic opportunities for patients with low sperm count (i.e. primary testicular dysfunction) or with altered sperm motility.

Cystic fibrosis phenotype evaluation and paternity outcome in 50 males with congenital bilateral absence of vas deferens.

BACKGROUND: Most infertile males with congenital bilateral absence of vas deferens (CBAVD) carry mutations on the cystic fibrosis transmembrane conductance regulator gene and may express mild cystic fibrosis (CF) symptoms. Barriers to paternity for these men can now be overcome by assisted reproduction. Our aims were to investigate the CF-related phenotype and clinical outcome for 50 patients with CBAVD seen at a CF adult centre between 1992 and 1999. METHODS AND RESULTS: The investigation of the patients included screening for 22 CF mutations and identification of the poly-T variant of intron 8, sweat testing, clinical investigation for CF-related extra-genital manifestations, and genetic counselling. CFTR mutations were detected on 56 alleles of the 50 patients. A total of 15 (30%) was compound heterozygote and 26 (52%) heterozygote. In all, 38% of the patients had a positive sweat test. Four patients were diagnosed with typical CF not detected previously. Twenty-one patients became fathers following ICSI (eight cases), artificial insemination by donor or IVF with sperm donor (seven cases) or through adoption (six cases). A mail survey allowed the identification of CF-related clinical symptoms. Information on the occurrence of CF-related symptoms was obtained for 58.5% of patients: in the absence of initial symptoms, no new clinical signs were reported. CONCLUSION: Patients diagnosed with CBAVD need genetic counselling before assisted reproduction. Even when no wish for paternity is expressed, CF gene screening should be associated with at least a sweat test and clinical evaluation because of possible mild forms of CF disease. Medical follow-up did not reveal any new symptoms.

Seminal transforming growth factor-beta in normal and infertile men.

Transforming growth factor-beta (TGFbeta) is a cytokine with autocrine and paracrine action in the testis and potent immunoregulatory and anti-inflammatory activities. In the present study, we examined the concentration of latent (acid-activatable) and free (active) TGFbeta in seminal plasma from normal subjects (n = 23) and infertile (n = 40) patients, by using a TGFbeta specific immunoenzymological assay, and a bioassay (CCL64 cell line growth inhibition) detecting any form of TGFbeta. Free TGFbeta1 was present in normal subjects at a concentration (1.82 +/- 1.06 ng/ml) close to that known to give maximal stimulation in vitro. In pathological groups, the mean concentrations were not significantly different from the normal ones. Latent TGFbeta1 was present in normal seminal plasma at a high concentration (92.4 +/- 29.2 ng/ml). In subjects with pathologies of both testis and genital apparatus, or with epididymal occlusion, mean latent TGFbeta1 concentrations were normal, whereas transferrin concentrations were lower. The concentrations found in the epididymal occlusion group indicate that TGFbeta1 is, for a large part, secreted by the genital tract. In the testicular pathology group, TGFbeta1 concentrations were 130.7 +/- 61.2 ng/ml, a mean not statistically different from normal, although higher. No differences were found between patients with high and normal blood plasma follicle stimulating hormone, and this is consistent with the notion that most TGFbeta1 in seminal plasma is not of testicular origin. The TGFbeta bioassay ensured that immunologically detected TGFbeta was present in a bioactive or bioactivatable form. Furthermore, the values found in normal and pathological seminal plasmas were usually higher than those detected by the immunoassay, suggesting that other forms of TGFbeta might be present. Together, the present data show that very large amounts of TGFbeta are present in human seminal plasma. The TGFbeta ligand assay in the seminal plasma appears to indicate no differences between normal and infertile subjects.

[Agenesis of the vas deferens in male infertility. A tentative classification based on 39 cases]. / Les absences de canaux déférents dans l'infertilité masculine. Tentative de classification à partir de 39 cas.

OBJECTIVES: We attempted to establish a classification of vas deferens agenesia observed in male infertility to better ascertain the pathophysiology involved and help guide genetic counselling. PATIENTS AND METHODS: Among 387 men consulting for infertility, agenesia of the vas deferens was confirmed by transrectal ultrasonography in 39. A search for 13 different cystic fibrosis mutations was performed in subjects without associated renal agenesia. RESULTS: Among the 39 cases, we identified 4 ultrasonographic categories: unique bilateral agenesia (25 cases), bilateral agenesia associated with renal agenesia (1 case), unique unilateral agenesia (6 cases) and unilateral agenesia with renal agenesia (7 cases). Presence or absence of a seminal vesicle was variable. A cystic fibrosis mutation was observed in 64% of the bilateral cases and in none of the unilateral cases. The delta F 508 mutation accounted for 62% of the mutations. Phenotypically, there was not distinction between patients with and without a mutation. In cases of unilateral agenesia of the vas deferens, azoospermia was more frequent in unique forms than in forms with associated renal agenesia. CONCLUSION: Presence or absence of a mutation does not affect the pathophysiology of vas deferens agenesia in cases without associated renal agenesia. Genetic counselling should however take into account the presence of an associated mutation. Vas deferens agenesia associated with renal agenesia is not associated with cystic fibrosis mutations and results from a different pathogenic mechanism.

[Male infertility caused by bilateral agenesis of the vas deferens: a new clinical form of cystic fibrosis?]. / Stérilité masculine par agénésie bilatérale des canaux déférents: une nouvelle forme clinique de mucoviscidose?

Congenital bilateral absence of vas deferens causes male excretory infertility and represents 1 to 2% of male infertility. Because of a genotypic similarity with cystic fibrosis, the possible in vitro fertilization with epididymal sperm requires careful genetic counselling. We studied genotype, sweat chloride concentration, respiratory function tests, sinus abnormalities, pancreatic and hepatic functions in 22 subjects with congenital bilateral absence of vas deferens. Among them, four were compound heterozygotus, all of them with the R117H mutation. Ten had a positive sweat test, one of them also being compound heterozygotus. Congenital bilateral absence of vas deferens and double mutation or positive sweat test led to high probable cystic fibrosis diagnosis in 13 subjects. Six subjects were heterozygotus for one cystic fibrosis mutation, criterium which is not sufficient for cystic fibrosis diagnosis; five of them had sinus abnormalities, present in 11 of the 22 subjects. Only three patients had no mutation nor sweat chloride abnormalities. This work confirms the high frequency of cystic fibrosis mutations in males with congenital bilateral absence of vas deferens, with a higher frequency of positive sweat test than in other publications, and a high frequency of sinus abnormalities. This monosymptomatic phenotype of cystic fibrosis suggests new hypotheses for a relationship between genotype and phenotype.

Diagnostic criteria for cystic fibrosis in men with congenital absence of the vas deferens.

The high frequency of cystic fibrosis (CF) mutations in males with absence of vas deferens supported the hypothesis of a primarily genital phenotype of CF disease. To consider the idea of an attenuated form of CF, we investigated 14 men with congenital bilateral aplasia of the vasa deferentia. All patients were consulting for infertility and none was known to have CF. The median age was 30.5 years (range, 20-38 yr). DNA analysis for 22 CF mutations showed at least 1 mutation in 10 patients (71%), whereas the CF carrier frequency is only 4% in the general population. Three compound heterozygotes were identified, all carriers of the R117H mutation. The sweat test was considered positive in 6 patients (43%), and a high frequency of radiologic evidence of sinus disease (8 patients) and of elevated antibodies to Pseudomonas (8 patients) was found. Only 2 patients were free of all these criteria for CF disease. This study strengthens the hypothesis that absence of vas deferens is an attenuated form of CF. We propose a combination of tests including DNA study, computerized tomographic scan of the paranasal sinuses, and testing of anti-Pseudomonas antibodies when the sweat test is inconclusive.

Detection of t(X;Y) in 2 XX males using fluorescent in situ hybridization.

Males with a 46,XX karyotype generally have Y chromosomal material translocated to the pseudoautosomal region of the X chromosome. We have delineated two such cases using two color fluorescent in situ hybridization with probes from the short arm (DYZ2), centromere (DYZ3), and long arm (DYZ1) of the Y chromosome and a centromeric probes for the X chromosome (DXZ1). Using these techniques, the two patients are identified as having the karyotype 46,X,der(X)t(X;Y) (p22;p11) and a phenotype consistant with this translocation. Azoospermia in these patients is explained by the absence of Y long arm material including the recently identified candidate gene family for spermatogenesis.

[Is male infertility caused by congenital bilateral atrophy of the vas deferens a clinical form of mucoviscidosis?]. / La stérilité masculine par atrophie congénitale bilatérale des canaux déférents est-elle une forme clinique de mucoviscidose?

We investigated clinical data, sweat electrolytes and cystic fibrosis (CF) mutations in twelve patients with congenital bilateral aplasia of vasa deferentia (CBAVD) to debate arguments for diagnosing CF. Sweat chloride concentration was definitely raised in four patients. Three patients are CF compound heterozygotes. Six patients are CF heterozygotes. This result reinforces the hypothesis that white males with CBAVD might have a mild form of CE.

Seminal alpha-glucosidase activity as a marker of epididymal pathology in nonazoospermic men consulting for infertility.

Glucosidase (alpha G) activity was measured in sperm free seminal plasma from 1200 patients consulting for primary infertility, in whom clinical examination of epididymides revealed some abnormalities and histories of genital infections. They constituted the group with epididymal pathology (P) that was compared with a reference group (R) of 246 men without any epididymal pathology. The distribution of alpha G was significantly different between the two groups, even if we considered only the subjects in group P with normal sperm count (PN: 353 men: p less than 10(-6). 15.9% of subjects in group PN exhibited alpha G values as low as vasectomized men, versus 1.2% in group R. A linear relationship was established between alpha G and sperm content in both groups, but alpha G activities were systematically lower in group P (y = 0.19 x + 64) than in group R (y = 0.30 x + 86). There was no correlation between alpha G and the percent of sperm motility. On the contrary, we found statistically more clinical epididymal abnormalities in cases of decreased alpha G activity than in cases of normal alpha G activity (p less than .01).

HLA antigens and infertility with sperm autoantibodies in man.

Vasectomized men with sperm auto-antibodies (S.A.A.) and high frequency of HLA-A20 antigen (Ag) were observed by Law et al. (1979); Hancock et al. (1983) observed the prevalence of HLA-A28 in infertile men with S.A.A. HLA-A, B and DR Ag were determined in a population of 80 Caucasian infertile men with (n = 22) or without (n = 58) agglutinating and/or cytotoxic S.A.A. in serum (S) and/or seminal plasma (SP), and the association between HLA-determinants and infertility with S.A.A. was investigated. The frequence of HLA-A28 Ag was found higher in the group with S.A.A. than without (18.2% vs 0.0%) but not statistically significant (chi 2 = 8.87; pc less than 0.15). The same result was obtained with HLA-B13 Ag (chi 2 = 8.87; pc less than 0.32). Lastly, there was no prevalence of HLA-DR Ag in men with S.A.A. In conclusion, no association was found between HLA-A, B and DR Ag and infertility with S.A.A. Nevertheless, two HLA-Ag were more frequently observed in the group with S.A.A.

[Etiologic evaluation of oligo-astheno-teratospermia]. / Bilan étiologique des oligo- asthéno- tératospermies.

The numerous etiological factors of male hypofertilities and the lack of precision relating to physiopathological mechanisms explain the weak therapeutic armamentarium. It is therefore absolutely necessary to perform a clinical, biological and paraclinical evaluation, logical and standardized, in order to determine etiology which can be cured and attempt a prognosis. The interview, the clinical examination and the sperm count represent the basis from which additional explorations (hormonal assays, biochemistry of the semen, immunological work-up, caryotype, paraclinical examinations) may be requested and provide more specific data.

Inhibition of spermatogenesis in men using various combinations of oral progestagens and percutaneous or oral androgens.

Eight men (experiment 1) requesting male contraception received a daily oral dose of 20 mg medroxyprogesterone acetate (MPA) combined with 125 mg percutaneous dihydrotestosterone (DHT). Three months later the mean sperm count was only diminished slightly; the replacement of DHT for four men by percutaneous testosterone at the same concentration led to a dramatic fall in sperm count. For 6-18 months all men were treated with MPA plus percutaneous testosterone (250 mg daily). The latter dose restored physiological levels of plasma testosterone. Follicle-stimulating hormone levels were inhibited more severely than in the DHT-treated group, whereas LH levels were variable. Azoospermia was achieved and maintained in six cases; two men were oligozoospermic and in one case a moderate secondary rise in the sperm count was observed. Twelve volunteers (experiment 2) received a daily oral dose of either 5 or 10 mg norethisterone acetate plus percutaneous testosterone (250 mg daily). All of them achieved azoospermia within 2 months, but two subjects later exhibited a partial restoration in sperm count. Follicle-stimulating hormone and LH levels were inhibited more severely than in the first experiment. The sperm count and gonadotrophin levels returned to initial values within 6 months after cessation of the treatment in both experiments. No side-effects were noted concerning blood parameters, libido or body weight. However, several female partners had elevated levels of plasma testosterone. In experiment 3 (13 volunteers), percutaneous testosterone was replaced by oral testosterone undecanoate (160 mg daily). Only seven men were azoospermic and most of them had lowered levels of plasma testosterone. Thus, the combination of percutaneous testosterone and oral progestagens appears to be the most convenient for male hormonal contraception.

PIP: In 3 successive experiments, the efficacy of various combinations of progestagens and androgens for male contraception was tested and compared. In the 1st experiment, 8 men received a daily oral dose of 20 mg medroxyprogesterone acetate (MPA) combined with 125 mg percutaneous dihydrotestosterone (DHT). After 3 months of this regimen, the mean sperm count was not significantly reduced. Thus DHT was replaced with percutaneous testosterone (250 mg/day) for 6-18 months. This move resulted in a dramatic fall in sperm count. Azoospermia was achieved in 6 cases and 2 men were oligozoospermic. In the 2nd experiment, 12 male volunteers received a daily dose of either 5 or 10 mg norethisterone acetate and 250 mg percutaneous testosterone. Azoospermia was achieved within 2 months by all 12 subjects, but 2 men later demonstrated a partial restoration in their sperm count. This regimen inhibited follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels more severely than was the case in the 1st experiment. In the 3rd experiment, percutaneous testosterone was replaced by 160 mg/day of oral testosterone undecanoate. Only 7 of the 13 volunteers in this experiment achieved azoospermia and most had lowered levels of plasma testosterone. The sperm count and gonadotropin levels returned to initial values within 6 months after cessation of treatment in these experiments and there were no significant side effects in terms of blood parameters, libido, or body weight. Overall, this research indicates that a combination of progestagens and androgens administered daily without injections induces satisfactory inhibition of spermatogenesis. If a fully active, nontoxic oral androgen were to become available as a substitute for percutaneous administration, hormonal male contraception could achieve more widespread acceptability.

[Silastic spermatocele]. / Spermatocèle en silastic.

The study was carried out in two stages. An initial experimental study in the rat showed that it was possible to attach a spermatocele to the rat epididymus and to collect spermatozoa during the months that followed. The clinical trial involved the implantation of 8 spermatoceles in 5 male patients. The technic is simple, as predicted by the animal study but, as previously described by numerous other authors, the communication between epididymus and spermatocele is obliterated within several months. These findings confirm the possibility of use of a spermatocele that is well tolerated and of simple functioning, but which does not allow fertile spermatozoa capable of producing pregnancy to be obtained.

[Comparative study of epididymal disorders in two populations of male patients consulting for infertility: one from the Maghreb region of North Africa and the other of European origin]. / Etude comparative des pathologies epididymaires dans deux populations d'hommes consultant pour infecondite: l'une maghrebine, l'autre europeene.

PIP: The authors carried out a retrospective study of 162 cases of male infertility explored in a hospital unit in Lyon, France. Assays of 1 -alpha-1,4-glucosidase (epididymal function marker) backed up by clinical findings were used to select 3 types of epididymal malfunction. 1) There was complete obliteration of the epididymal duct, resulting in azoospermia. This diagnosis was based on both testicular biopsy findings, demonstrating unimpaired spermatogenesis and on the dramatically reduced level of assayed activity ( 40 mIU/ejaculation), as well as on clinical findings. 2) There was anamalous epididymal function combined with moderate oligoasthenozoospermia or normospermia. In these cases, low levels of assayed activity do not parallel fairly high sperm counts (between 20-30 million spermatozoal/ml). 3) There were those cases which were difficult to interpret and which involved severe oligoasthenozoospermia ( 5 million/ml) and reduced level of epididymal marker, suggesting partial blockage of the epididymis due to a focus of infection. Varicoceles were found more frequently among the European population, whereas a history of genital infection was more frequent among the North African population. However, when the various types of abnormality in the spermatogram were related to patient history and epididymal abnormality, no differences were found between the 2 populations. (author's modified)^ieng

Alpha-glucosidase as a specific epididymal enzyme marker. Its validity for the etiologic diagnosis of azoospermia.

Azoospermic semen was obtained from 39 vasectomized men and 93 patients consulting for infertility. The latter underwent bioclinical investigations including measurement of plasma FSH and testicular biopsies. Carnitine content and alpha-glucosidase activity in semen samples were measured. The activity of alpha-glucosidase was determined systematically by a semiquantitative microtechnique and was verified by a spectrophotometric assay. A positive correlation was observed between carnitine and alpha-glucosidase activity. Both parameters were severely diminished in semen from the vasectomized men and the patients suffering from a complete obstruction of the genital tract. Enzyme activity was the most discriminant parameter in terms of sensitivity and specificity. The measurement of alpha-glucosidase in semen is a simple and sensitive method for determining the origin of azoospermia when used in conjunction with assays for plasma FSH.

[Role of scanography in the diagnosis of parathyroid adenomas. Apropos of 27 cases]. / Place de la scanographie dans le diagnostic des adénomes parathyroïdiens. A propos de 27 cas.

Scanner imagine pre-operatively in 27 patients with hyperparathyroidism provided assessment of the sensitivity (92 p.cent) and diagnostic precision (92.5 p.cent) of this investigational method. The need for a strict methodology and the advantages gained by high resolution and angioscanning programmes are emphasized. After analyzing the results, a review of the published literature enables the advantages and the limits of the scanner X to be defined, and a strategy for exploration of parathyroid adenomas to be proposed.