Phenotypic variation within European carriers of the Y-chromosomal gr/gr deletion is independent of Y-chromosomal background.

BACKGROUND: Previous studies have compared sperm phenotypes between men with partial deletions within the AZFc region of the Y chromosome and non-carriers, with variable results. In this study, a separate question was investigated, the basis of the variation in sperm phenotype within gr/gr deletion carriers, which ranges from normozoospermia to azoospermia. Differences in the genes removed by independent gr/gr deletions, the occurrence of subsequent duplications or the presence of linked modifying variants elsewhere on the chromosome have been suggested as possible causal factors. This study set out to test these possibilities in a large sample of gr/gr deletion carriers with known phenotypes spanning the complete range. RESULTS: In total, 169 men diagnosed with gr/gr deletions from six centres in Europe and one in Australia were studied. The DAZ and CDY1 copies retained, the presence or absence of duplications and the Y-chromosomal haplogroup were characterised. Although the study had good power to detect factors that accounted for >or=5.5% of the variation in sperm concentration, no such factor was found. A negative effect of gr/gr deletions followed by b2/b4 duplication was found within the normospermic group, which remains to be further explored in a larger study population. Finally, significant geographical differences in the frequency of different subtypes of gr/gr deletions were found, which may have relevance for the interpretation of case control studies dealing with admixed populations. CONCLUSIONS: The phenotypic variation of gr/gr carriers in men of European origin is largely independent of the Y-chromosomal background.

Immune system parameters in children of Central and Eastern Europe: the CESAR study.

OBJECTIVE: of this paper is to compare observed values of immune parameters obtained in the CESAR study (The Central Europe Study of Air Pollution and Respiratory Health, funded by EC PHARE program) with ranges derived from other large population-based studies. STUDY DESIGN: Data were collected in healthy school children aged 9-11 years, in 6 countries: Bulgaria, the Czech Republic, Hungary, Poland, Romania and the Slovak Republic with the same standard approach in 1996. Random samples of 85 children per country, from 19 communities were selected from children having completed the health questionnaire, in total 495 children were analyzed. Lymphocyte subsets were determined by two-colour flow cytometric immunophenotyping using the lysed whole blood method (Becton-Dickinson). For determination of immunoglobulin concentration in sera nephelometric method (Behring Nephelometer system) was used. RESULTS: Medians, (5th-95th percentiles) of the lymphocyte subsets absolute count (x 10(9)/l) were as follows: CD19+ B cells 0.36 (0.13-0.66), total CD3+ T cells 1.74 (0.98-2.90), CD3+CD4+ helper-inducer T cells 0.95 (0.47-1.78), CD3+CD8+ suppressor/cytotoxic T cells 0.71 (0.38-1.22), CD3-CD16+56+ NK cells 0.36 (0.14-0.78), and for CD3+CD4+/CD3+CD8+ ratio 1.4 (0.8-2.4). Medians, (5th-95th percentiles) of percentages of lymphocyte subpopulations (%) were as follows: CD19+ B 13 (7-22), CD3+ T 70 (59-80), CD3+CD4+ T 38 (27-48), CD3+CD8+ T 28 (20-39), CD3-CD16+56+ NK cells 14 (6-27). Medians, (2.5th-97.5th percentiles) of the total immunoglobulin [g/l] were 11.7 (7.4-18.2) for IgG, 1.2 (0.5-2.5) for IgM, and 1.5 (0.5-3.4) for IgA. Based on the aspects of the size of the CESAR immune biomarker study and on the use of the standardized protocols we recommend to use the reference ranges on lymphocyte subsets and immunoglobulin in Europe as provided by this study.

Reversible meiotic abnormalities in azoospermic men with bilateral varicocele after microsurgical correction.

Because of a possible relationship between microenvironmental disturbances and meiotic abnormalities and of a straight relationship between lower-quality semen in patient carrying a varicocele and first meiotic non-disjunction, bilateral bipolar testicular biopsies are realized according the thermic differential gradient described in varicocele. Systematic meiotic studies of multiple testicular biopsies from 65 azoospermic men with bilateral varicocele were done in a multi-centric study on microsurgical correction of bilateral varicocele with microthermic intra-operative evaluation using minimally invasive thermal microsensors (Betatherm 10K3MCD2). In the present study abnormal temperature raising, histomorphometric abnormalities (spermatocyte arrest) and meiotic abnormalities (class IIC) are strongly correlated. In the ten patients submitted to another testicular biopsy procedure six months after surgery for TESE, normal thermal differential is registered and no meiotic abnormalities recurrences are found.

Aetiology of haemospermia.

There are several unknown factors which cause haemospermia. An earlier developed diagnostic scheme has been expended by novel imaging techniques and biopsy methods. A detailed case history, physical examination and microscopic analysis of the ejaculate is required. In haemo-pyospermia a complete microbiological analysis must be escalated. Noninvasive imaging techniques (ultrasound, computer tomography and magnetic resonance imaging) help in detecting calculous and malignant diseases. So far, as a precise diagnosis has not been available, urethroscopy has been performed. Malignancies (prostate, seminal vesicles) must be histologically verified by biopsies. In contempt of our efforts the practice shows a part of haemospermia remaining essential. Analysing two time periods we found prostatic calculi, chronic prostatitis and carcinoma of the prostate unequivocally as most frequent causes. Considering the rare genital malignancies we find more than 10% frequency. Notably, in our study only 2.4% of the malignancies occurred in patients under 40 years of age. Hence a detailed diagnosis is advocated in haemospermia patients over 40 years. Finally, we may state that in contempt of the applied modern imaging techniques 15% of patients with haemospermia had unknown aetiology.

Five years experiences with microinjection of testicular spermatozoa into oocytes in Hungary.

The aim of the study was to summarize our five years experience (1996-2000) of testicular spermatozoa for intracytoplasmic sperm injection in Hungary. The influence of sperm count, maternal age, number of transferred embryos, and application of assisted hatching on outcome was investigated. Testicular spermatozoa were retrieved by microsurgical testicular sperm extraction. Samples were classified depending on the number of spermatozoa. Indication for testicular sperm extraction in conjunction with intracytoplasmic sperm injection was severe azoospermia or azoospermia combined with tubal origin infertility. Ovarian stimulation was carried out using an ultrashort protocol with GnRH agonist and gonadotrophin. Intracytoplasmic sperm injection was performed without PVP. Embryos were cultured for 48 or 72 h before embryo transfer. Indications for assisted hatching included elevated maternal age, increased zona thickness or at least two previous unsuccessful IVF cycles. Testicular spermatozoa were successfully retrieved in 218 out of 273 cases. Extreme low sperm count was found more frequently in cases of nonobstructive azoospermia. No significant differences were observed in fertilization rate (61.1% vs. 51.7%) or clinical pregnancy rate (29.0% vs. 26.7%) between patients with obstructive or nonobstructive azoospermia. Maternal age, number of transferred embryos and application of assisted hatching had a significant effect on outcome. A total of 55 clinical pregnancies were achieved, including 14 sets of twins, three sets of triplets and two sets of quadruplets. It is concluded that testicular sperm extraction is an efficient way of obtaining testicular spermatozoa, allowing not only successful fertilization by ICSI, but also freezing of testicular spermatozoa for use in subsequent cycles.

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.

Immune biomarkers in relation to exposure to particulate matter: a cross-sectional survey in 17 cities of Central Europe.

Human population data on air pollution and its effects on the immune system are scarce. A survey was conducted within the framework of the Central European Study of Air Quality and Respiratory Health (CESAR) to measure a panel of immune biomarkers in children of Bulgaria, Czech Republic, Hungary, Poland, Romania, and Slovakia. Seventeen cities were chosen to represent a wide range of exposure to outdoor air pollution. In each, ambient particulate matter of less than 10 microns diameter and less than 2.5 microns diameter (PM10 and PM2.5) were measured with a Harvard impactor. Blood was collected from 366 school children aged 9 to 11 yr between 11 April and 10 May 1996. The percentage of B, total T, CD4+, CD8+, and natural killer (NK) lymphocytes was determined by flow cytometry (Becton Dickinson); total immunoglobulins of class G, M, A and E (IgG, IgM, IgA, and IgE) were measured in serum using nephelometry (Behring). Associations between PM and each log-transformed biomarker concentration were studied by linear regression, in a two-stage model. The yearly average concentrations varied from 41 to 96 micrograms/m3 for PM10 across the 17 study areas, from 29 to 67 micrograms/m3 for PM2.5, and from 12 to 38 micrograms/m3 for PM10-2.5 (coarse). Number of B, CD4+, CD8+, and NK lymphocytes increased with increasing concentration of PM, having adjusted for age, gender, parental smoking, laboratory of analysis, and recent respiratory illness. Differences in lymphocyte number were larger and statistically significant for exposure to PM2.5. Similar results were found when we examined the association between PM and lymphocyte number separately for each laboratory. Total IgG was increased with increasing concentration of PM, significantly in the case of PM2.5. When we repeated the analyses with two other statistical approaches the results did not differ from those reported here. The effect of coarse PM on lymphocyte numbers appears small in comparison to PM2.5. One possible interpretation of our findings is that long-term exposure to airborne particulates leads to inflammation of the airways and activation of the cellular and humoral immune system.

A high frequency of Y chromosome deletions in males with nonidiopathic infertility.

Microdeletions of the long arm of the human Y chromosome are associated with spermatogenic failure and have been used to define three regions of Yq (AZFa, AZFb, and AZFc) that are recurrently deleted in infertile males. In a blind study we screened 131 infertile males (46 idiopathic and 85 nonidiopathic) for Y chromosome microdeletions. Nineteen percent of idiopathic males, with an apparently normal 46,XY chromosome complement had microdeletions of either the AZFa, AZFb, or AZFc region. There was no strict correlation between the extent or location of the deletion and the phenotype. The AZFb deletions did not include the active RBM gene. Significantly, a high frequency of microdeletions (7%) was found in patients with known causes of infertility and a 46,XY chromosome complement. These included deletions of the AZFb and AZFc regions, with no significant difference in the location or extent of the deletion compared with the former group. It is recommended that all males with reduced or absence sperm counts seeking assisted reproductive technologies be screened for deletions of the Y chromosome.

Testicular biopsy helping assisted reproduction.

Authors have performed testis biopsy on 21 male patients according to the predesigned protocol. The biopsy has been carried out from scrotal excision, with the exposure of both testis with microsurgical method. The results have been evaluated according to the spermogram groups. Correlation of FSH values and histological appearance of both testis was analyzed. Their method and experiences have been evaluated the effectiveness of assisted reproduction.

[The combined occurrence of pubertal gynecomastia and varicocele]. / Das gemeinsame Vorkommen der Pubertätsgynäkomastie und der Varikozele.

Examinations of 39 men between 14 and 18 years of age revealed that half of the cases of pubertal gynaecomastia are false gynaecomasties. In reality, the patients are suffering from fibrous mastosis. It is shown that gynaecomastia is mostly accompanied by left-sided varicocele, whereas in varicocele complaints there are only few cases of gynaecomastia. It is concluded that in pubertal gynaecomastia it is necessary to determine whether the disease is merely a temporary fibrosis that will heal by itself, or whether it is a sign of some other, graver disease. On the other hand, the state of the testicles and the scrotum should be considered in every case. Surgery of pubertal gynaecomastia is considered unnecessary; however, invasive treatment of the varicocele is very important. The importance of psychological care of the patients is stressed.