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A study on the reproductive toxicity of erythrosine in male mice.

Worldwide usage of different colouring agents in the food industry prompted us to study their toxicity. The potential adverse effects of erythrosine (ER, FD & C Red No. 3) on the spermatogenesis process were investigated in adult mice. Testicular lactic dehydrogenase isoenzyme activity (LDH-X), a pachytene spermatocyte marker of testicular toxicity, was significantly decreased to 71.8% and 68.6% of the control value after daily p.o. administration of ER (21 days) in doses of 68 and 136 mg kg-1 respectively. At the same time, the normal average epididymal sperm count as well as the percentage of motile sperms were significantly inhibited by about 50% and 57% respectively. Moreover, ER was shown to disrupt the normal morphology of the sperm head. Thus, after 5 daily p.o administrations of ER in doses of 680 and 1360 mg kg-1 (equivalent to 10 and 20% of its LD50) it increased the incidence of sperms with abnormal head by about 57% and 65% respectively. The induced increase in sperm abnormalities could enhance the spermatogenic dysfunction and germ cell mutagenicity. These findings indicate that ER in the used doses has a potential toxic effect on spermatogenesis in mice and in turn, it may affect its testicular function and reproductive performance.

Animals↗

Reproductive effects of chronic administration of murine interferon-gamma.

Daily subcutaneous doses of 0.02, 0.2, or 2 mg/kg/d of recombinant murine interferon-gamma (rmuIFN-gamma) were given to mice on postnatal days 8 through 60 to determine effects on maturation, behavioral/ functional development, and reproductive capacity. Male mice receiving 2 mg/kg/d rmuIFN-gamma had delayed sexual maturation, reduced epididymal and testes weights, reduced sperm count and concentration, and sperm abnormalities (crimped flagellum). Mating performance and fertility were also reduced in the absence of altered histopathology of the testes. Males given 0.2 and 2 mg/kg/d had swelling and ulcerative dermatitis around the urogenital area, which were observed after sexual contact and attributed to a bacterial infection. Motor activity (time spent in movement) was decreased in all mice receiving 2 mg/kg/d. No microscopic changes observed in any organs were attributed to rmuIFN-gamma administration.

Analysis of Variance↗

A low sperm concentration does not preclude fertility in men with isolated hypogonadotropic hypogonadism after gonadotropin therapy.

In order to define the minimal number of sperm needed for conception, we studied semen characteristics of men with isolated hypogonadotropic hypogonadism (IHH) who became sperm-positive during gonadotropin therapy. Twenty-two of 24 men (92%) proved fertile, initiating a total of 40 pregnancies. The mean (+/- standard error of the mean) sperm concentration at the time of conception was 16.7 +/- 4.0 X 10(6)/ml. However, 71% of pregnancies were conceived when the mean sperm concentration was less than 20 X 10(6)/ml; in 16%, the mean sperm concentration was less than 1 X 10(6)/ml. Mean total sperm count correlated highly with sperm concentration (r = 0.67, P less than 0.001). We conclude that men with IHH can initiate conception even when their sperm concentration is well below the conventional lower limit of 20 X 10(6)/ml.

Adolescent↗

Subclinical varicocele: the effectiveness of varicocelectomy.

To investigate the controversial concept that subclinical varicoceles are detrimental to spermatogenesis, we undertook a prospective study of the effectiveness of surgery in 56 infertile men with clinical and subclinical varicoceles. Our statistical approach specifies improvement as a change from individual baseline level. The over-all variability in sperm counts precluded the demonstration of significant changes in sperm density consequent to surgery. Improvement in sperm motility was seen in both groups, being slightly greater in the subclinical group. The changes were inversely related to baseline levels and increased with time from surgery. Therefore, it appears that the detection of subclinical varicoceles may be warranted in infertile men with abnormal semen parameters.

Humans↗

Flow cytometric evaluation of sperm from patients with testicular carcinoma.

Semen samples from 14 patients with testicular cancer were analyzed by new flow cytometry techniques and by conventional semen analysis. Samples were obtained post-unilateral orchiectomy and prior to further treatment. With only 1 exception, light microscopic analysis of sperm count and morphology indicated a decrease of normal sperm production and incomplete or abnormal differentiation. Flow cytometry of semen aliquots showed, with the same exception as above, increased acridine orange staining of the sperm nuclear chromatin, attributed to abnormal chromatin composition and condensation. In addition, the chromatin structure of isolated sperm nuclei was abnormally sensitive to thermal stress. These changes were present up to 15 months post orchiectomy and suggest abnormal spermiogenesis in the contralateral testis of patients with testicular tumors. Of the 14 patients studied only 1 fell well within the normal range for all parameters measured; 2 other patients were in the low normal range for all parameters and 1 patient fell below normal for light microscope derived measurements but was normal by flow cytometry. The other 10 patients had abnormal measurements for all parameters. The assessment of spermiogenesis by flow cytometry studies of sperm chromatin appears to be a sensitive and valuable new parameter of fertility that cannot be determined by conventional semen analysis.

Acridine Orange↗

Vasovasostomy and semen quality.

During a recent 8-year 4-month period 205 men underwent bilateral vasovasostomy. Of those patients for whom followup information was available 43 per cent have produced a pregnancy. Semen analyses after vasovasostomy revealed good sperm counts for the most part but generally reduced sperm motility and viability. Efforts are underway to evaluate whether the defect may be the result of abnormal epididymal function or of sperm agglutinating or immobilizing antibodies.

Adult↗

Comparison of the Sperm Quality Analyzer IIC variables with the computer-aided sperm analysis estimates.

Sperm Quality Analyzer (SQA) IIC, an upgrade version, is an inexpensive device and provides a quantitative estimation of sperm motility, whereas the use of computer-aided sperm analysis (CASA) provides high precision and provision of quantitative data on sperm kinetics. The aim of the present study was to evaluate if the SQA IIC variables correlated with the CASA estimates. Semen quality analysis of 71 fresh semen samples was performed using SQA IIC and CASA. Total sperm concentration, percentage of progressively motile sperm, percentage of normal morphology, motile sperm concentration, sperm motility index (SMI) and functional sperm count (FSC) determinations were performed using SQA IIC. Sperm concentration, sperm motility, and sperm motion variables including amplitude of lateral head displacement (ALH), beat cross frequency (BCF), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN=VSL/VCL), and straightness (STR=VSL/VAP) were evaluated simultaneously on the same semen samples using CASA. The sperm characteristics were compared between SQA IIC and CASA. There were significant correlations of sperm concentration (r=0.634, p < 0.0001), sperm motility (r=0.697, p < 0.0001), and motile sperm concentration (r=0.757, p < 0.0001) between the two devices. Both SMI and FSC significantly correlated with eight CASA estimates, including sperm concentration, sperm motility, motile sperm concentration, ALH, VCL, VSL, VAP, and Rapid. SQA IIC is simple and easy to use. Moreover, the SQA IIC variables well correlated with the CASA estimates. As a screening test for semen quality, SQA IIC is considered as useful in the management of male infertility.

Diagnosis, Computer-Assisted↗

Semen collection from mice: electroejaculation.

The effects of device type (electrostimulator, function generator or computer-generated waveforms), waveform (square, triangle or sine wave), probe type (ring or strip) and anaesthetic compound (ketamine/xylazine combination or pentobarbitone sodium) were investigated on electroejaculation (EEJ) responses of C57B1 x CBA and C57Bl/6J mice. Ejaculates were analysed for total sperm count and motility variables using computer-assisted sperm analyses. Automated computer-generated waveforms delivered through a sound card were more effective and reproducible compared with waveforms generated by function generator and electrostimulator. Sine waves and triangle waves were found to be more effective in producing ejaculate than square waves. As an anaesthetic, pentobarbitone sodium tended to outperform ketamine/xylazine across waveforms and strains. Strip probes failed to produce any ejaculate regardless of the device or waveform employed. Sperm obtained by EEJ exhibited poor motility and C5B1/6J mice had lower motility variables than C57BI x CBA mice.

Anesthesia↗

Semen quality of 324 fertile Japanese men.

BACKGROUND: A number of studies have indicated regional differences in semen quality. To examine the current status in Japan, we undertook a cross-sectional study on the semen quality of fertile Japanese men for comparison with recent European results. METHODS: Semen parameters of 324 fertile men from the Kawasaki/Yokohama area were investigated. The semen parameters were compared with those published for fertile men from four European cities, Copenhagen, Paris, Edinburgh and Turku. RESULTS: When adjusting for confounders such as ejaculation abstinence period and age, the lowest sperm concentrations were detected in men from Kawasaki/Yokohama followed by men from Copenhagen, Paris, Edinburgh and Turku, but only the differences between men from Kawasaki/Yokohama and men from Edinburgh and Turku were significant (P=0.0008 and P<0.0001, respectively). Total sperm count, percentage of motile sperm and percentage of normal sperm observed in Kawasaki/Yokohama were significantly lower than those from all European centres except for motile sperm in men from Paris. CONCLUSIONS: Japanese fertile men had a semen quality at the level of Danish men, who have been reported to have the lowest among investigated men in Europe. The low level of semen quality of the fertile Japanese men may be due to lifestyle or other environmental factors; however, ethnic differences caused by different genetic variation or combinations cannot be ruled out by this study.

Ejaculation↗

Is smoking a risk factor for decreased semen quality? A cross-sectional analysis.

BACKGROUND: Previous studies suggest a deleterious effect of cigarette smoking on semen quality, but their results have not been consistent. We studied the association between current smoking and semen characteristics and hormonal levels in a large group of healthy men. METHODS: From 1987 to 2004, seven separate occupational or environmental semen quality studies were co-ordinated by our department. A total of 2562 men participated, each providing semen and blood sample and answering a questionnaire about lifestyle and factors related to health. Appropriate semen and smoking data were available for 2542 men. RESULTS: Adjusting for study, age and other covariates, we observed an inverse dose-response relation between smoking and semen volume, total sperm count and percentage motile sperm. Heavy smokers had a 19% lower sperm concentration than non-smokers. We found a positive dose-response relationship between smoking and testosterone, LH and the LH/free testosterone ratios. CONCLUSION: Current smoking in adult life moderately impairs the semen quality. It is well known that semen quality is associated to fecundity. Therefore, it would be sensible to advise men to abstain from smoking to avoid decreased fecundity.

Adult↗

Kallikrein and male subfertility. Usefulness of high-unit kallikrein tablets.

We studied the therapeutic effect of long-term oral administration of Kallikrein tablets on male subfertility. Subjects were 26 patients with poor sperm motility and divided into two groups according to the rate of motile spermatozoa: less than 40% (Group A: 9 cases) and 40% or more (Group B: 17 cases). Patients in Group A administered with 600 K.U./day orally for 6 months and those of Group B received 300 K.U./day for initial 3 months and 600 K.U/day for the ensuing 3 months. The changes of semen parameters during the treatment were as follows. a) No increase in sperm count. b) significant improvement of quantitative and qualitative sperm motility. c) Improvement of sperm morphology. d) Significant increase in total number of motile spermatozoa. e) Volume of semen was increased without statistical significance. Testosterone, FSH, and LH levels in blood plasma did not change significantly. The constituents of seminal plasma (fructose, cholesterol, ALP, ACP, GOT and LDH) were determined. Although ACP and GOT showed an increasing tendency and ALP tended to decrease during treatment, we found no biochemical parameter changing parallel with therapeutic results. Eight out of the 26 subjects successfully impregnated their wives (conception rate: 30.8%). No remarkable side effect was detected.

Administration, Oral↗

Quantitative studies of cytological damage in mouse testis produced by exposure to heat.

Damage in gonadal cells produced by exposure of the hindquarters of conscious male mice to water at temperatures up to 43 degrees C was studied by measuring the percentage of testicular cells which do not exclude trypan blue 4 h after exposure, sperm count up to ten wk later, and the percentage of morphologically abnormal sperm up to ten wk later. It was found that the percentage of cells stained with trypan blue was significantly increased by exposure for 0.5 h at 41 degrees C or 2 h at 37 degrees C. Sperm count and the percentage of abnormal sperm were affected by 0.5 h at 43 degrees C or 2 h at 40 degrees, but not by lower temperatures. Since there is a high incidence of abnormal sperm after some heat treatments, the use of this assay for mutagen screening should be approached with caution.

Animals↗

Predictive indicators of successful varicocele repair in men with infertility.

OBJECTIVE: We sought predictive indicators of functionally successful repair in 168 infertile Japanese males who underwent high ligation of a varicocele via a retroperitoneal approach. METHODS: Possible indicators evaluated included age at marriage, duration of infertility, testicular volume, varicocele grade, seminal analysis findings, and serum concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), and testosterone. All patients had palpable or visible unilateral left varicoceles. Success in conception was evaluated in all subjects 1 year after varicocele repair. RESULTS: No associations were observed between outcome and mean age at marriage, infertility duration, varicocele grade, seminal volume, or serum PRL and testosterone concentrations, but testicular volume, sperm count, percentage of motile sperm, and serum FSH and LH concentrations were significantly associated with outcome. Multiple regression analysis of dependence of outcome revealed testicular volume and serum FSH concentrations to be independent predictors. CONCLUSIONS: Varicocele repair is likely to accomplish fertility in patients with a combined testicular volume of at least 30 mL or serum FSH concentrations lower than 11.7 mIU/mL.

Adult↗

[The effect of methylmercury on testicular function of mice].

The results showed that the amount of methylmercury (MeHg) in testis correlated positively with exposure doses (r = 0.99 P less than 0.01). MeHg affected the process of spermatogenesis, causing decrease of sperm count and increase of sperm abnormalities. Pathological and electron microscopic examinations indicated that spermatogenic cells were damaged by MeHg especially spermatogonium and spermatocyte. Changes of microstructure were mainly degeneration, fragmentation vacuolation and large lipid drop formation of spermatogenic cells. The acrosome of many spermatids showed obvious changes. Destruction of blood-testis barrier was observed in 1/10 LD50 group. Quantitative histological examination also confirmed that MeHg resulted in lessening of its area of seminiferous tubules and reduction of germ cells. The contents of testosterone were not decreased in the treated groups other than the 1/10 LD50 group.

Animals↗

Physical, metabolic, and hormonal effects on men of long-term therapy with medroxyprogesterone acetate.

Twenty-three men were treated with injectable medroxyprogesterone acetate (MPA) for sex-offending behavior. The physical, hormonal, and metabolic effects were monitored. The patients had significant (P less than 0.001) weight gain and increases (P less than 0.0001) in systolic blood pressure independent of weight gain. MPA therapy decreased testosterone (P less than 0.0001), luteinizing hormone (P less than 0.005), and testicular size (P less than 0.001), but not follicle-stimulating hormone. MPA's suppression of total sperm count was not consistent, but MPA significantly reduced sperm motility (P less than 0.001) and percent normal morphology (P less than 0.005). Statistically insignificant increases in insulin response without changes in glucose tolerance occurred in all patients, but only one patient developed overt diabetes mellitus. Three men developed gallstones, but none had significant changes in liver function. Patients receiving high-dose MPA therapy should be carefully monitored for blood pressure changes, diabetes mellitus, and gallbladder dysfunction.

Adult↗

The clinical varicocele in infertility.

Of 238 infertile males seen over a 47-month period, 47 (20%) had a varicocele. Of these 51% had an abnormal sperm count (21% were azoospermic and 30% oligozoospermic) and 60% had less than a third of sperms motile; 23 patients were subjected to gonadotrophin assays. Serum follicle stimulating hormone and leutinising hormone levels were elevated in 48% and 70% of patients respectively. Serum prolactin assayed in 19 patients was raised in 10% and serum testosterone estimated in 16 patients was low in 6%. Twenty-five patients (53%) were advised to undergo surgery but only 10 agreed. Of these, 1 showed a dramatic improvement in his spermiogram and a pregnancy ensued. The sperm count and motility of 2 others improved after treatment with prostaglandin synthetase inhibitors, and 1 after testosterone. Medical treatment improved the count and motility in 7 of 8 patients in whom surgery was not suggested as primary treatment because of abnormal hormonal results. This study has shown that infertile patients with varicocele are a heterogeneous group and that improvement in spermiograms can best be achieved by selective treatment; this may be surgery, surgery followed by medical treatment, or medical treatment alone.

Follicle Stimulating Hormone↗

Testicular cancer and spermatogenesis.

We retrospectively analysed the characteristics of 54 men with testicular cancer. The group comprised 32 men with pure seminoma and 22 with non-seminoma germ cell tumours (NSGCT). This group was further compared to 190 healthy sperm donor candidates. Sperm quality was found to be higher in the seminoma versus the NSGCT patients in: sperm concentration [50 (0-230) versus 17 (0-288) x 10(6)/ml, P < 0.001], total motile sperm counts (TMC) [57 (0-508) versus 12 (0-854) x 10(6)/ejaculate, P = 0.002], post-thaw forward motile concentration [3 (0-28) versus 1.7 (0-17) x 10(6)/ml, P = 0.003] and motility percentage [20 (0-57) versus 12.5 (0-42) %, P = 0.002]. Serum hormone concentrations did not differ between these two sub-groups, although the follicle stimulating hormone concentrations were higher than normal in both (14.6 +/- 2.5 versus 10.4 +/- 1.4 mIU/ml, P > 0.05). As is well documented, cancer patients were found to have lower sperm quality compared to healthy candidates. The existence of these differences, and the fact that testicular cancer affects spermatogenesis, indicated that the mechanisms involved in the deterioration of sperm quality can, at least partially, be attributed to the type and origin of the malignant cancer. The higher sperm counts in the seminoma group may be related to the fact that the resemblance of the seminoma cells to normal germ cells is greater than that of the NSGCT cells, and therefore they retain a better capacity to function. Due to modern assisted reproductive technologies and micromanipulation achievements, the lower yield of spermatozoa in severe cases is no longer a major obstacle to offering cryopreservation to these patients.

Cryopreservation↗

Histological evaluation of multiple testicular biopsies helping assisted reproduction.

Because of the fall in population in Europe, the evaluation of causes of male infertility is becoming more and more important. Testicular biopsy is a useful method of examination in the area of assisted reproduction. Testicular biopsies of 85 infertile males were evaluated histologically. According to the sperm count, 31 patients had azoospermia, 22 oligo-astheno-teratozoospermia (OATS), 29 oligozoospermia, and 3 astheno-teratozoospermia. The biopsies were carried out by atraumatic microsurgery. Samples were taken from the upper inner and lower outer quadrant of both sides. Bouin-fixed, paraffin-embedded blocks were cut and HE slides examined. Samples taken from azoospermic patients showed atrophy, maturation arrest, normal histology or Sertoli cell-only syndrome. Oligozoospermic patients presented maturation arrest, hypospermatogenesis or normal histology. In the OATS group maturation arrest hypospermatogenesis and atrophy were found. The spermatogram groups showed significant correlation with the homogeneity of the spermatogenesis. The sperm count alone is not sufficient in assessing spermatogenesis. Four samples give more adequate information than just one, because spermatogenesis is not homogeneous in the testis. After evaluating diagnostic biopsies, tissue for TESE can be taken from the most appropriate location in the testis.

Adult↗