[Aneuploidy frequency in sperm of fertile men].

Analysis of sperm aneupoidy in 11 healthy men using two- or three-color FISH permitted to determine the average frequency of disomy in chromosomes 13 and 21 (0.11% and 0.2%, respectively), disomy in chromosome 18 (0.05%) and reveal gonosomal aneuploidy variants and their frequency. The frequency of XX disomy was 0.04%; XY, 0.17%; YY, 0.06%; and gonosomal nullisomy, 0.29%. We also assessed the frequency of meiotic nondisjunction of chromosomes 13, 21, 18, X, and Y and the frequency of XX, XY, and YY diploid chromosomes in sperm. The XY variant prevailed in gonosomal aneuploidy and diploidy and was associated with abnormal chromosomal segregation in meiotic anaphase I. The contribution of human sperm chromosomal imbalance to early embryonic lethality and to some chromosomal diseases of syndrome type in the offspring is discussed.

[Effect of herpes simplex virus on spermatogenesis].

To investigate the effect of Herpes Simplex virus (HSV) on spermatogenesis, HSV in ejaculate was detected by a rapid cultural method in 268 infertile males and 47 healthy ones. The number of mobile spermatozoa in HSV infected samples was less than in non-infected samples (21 mln/mlversus 40 mln/ml, p = 0.0001). The relative number of morphologically normal gametes was 13% versus 19% (p = 0.002), respectively. The quantitative karyological test discovered that males with HSV-infected ejaculate have more degenerating sex cells while in high virus contamination (more than 10 virus particles in 1 ml) the number of spermatides and spermatocytes of the 1 order at diploten stage is low. Organic testicular culture was used for more detailed study of pathogenetic mechanisms of HSV impact on spermatogenesis. Testicular explants infection was associated with reduction in the number of spermatogones, spermatocytes and spermatides on culturing week 2. The above findings reveal some pathogenetic mechanisms underling fertility disorders in males with HSV infection: a gametotoxic effect of the virus reducing populations of spermatogones, spermatocytes and spermatide; affected mobility and morphological characteristics of spermatozoa. Detection of the mechanisms of HSV action on spermatogenesis opens a perspective of antivirus drug administration in combined treatment of male infertility.

[The frequency and spectrum of mutations and the IVS8-T polymorphism of the CFTR gene in Russian infertile men].

The frequency and spectrum of mutations and the IVS8- T polymorphism of the CFTR gene have been studied in a sample of 963 in Russian infertile men. Mutations have been found in 48 out of 1926 analyzed chromosomes (2.5%) in the heterozygous state (n = 46) and in the compound heterozygote L138ins/N1303K (n = 1/n = 1). A CFTR gene mutation was combined with the 5T allele (mutCFTR/5T) in 11 patients. The following mutations have been found: F508del (n = 18), CFTRdele2,3 (21kb) (n = 9), W1282X (n = 7), 2143delT (n = 4), 3849+10kbC>T (n = 2), L138ins (n = 2), 1677delTA (n = 1), 2184insA (n = 1), 3821delT (n = 1), G542X (n = 1), N1303K (n = 1), and R334W (n = 1). The F508del mutation is the most frequent; it has been detected in 37.5% of the affected chromosomes. The total proportion of four mutations (F508del, CFTRdele2,3 (21kb), W1282X, and 2143delT) is about 79% of all mutations found. The 5T allele has been found in 10.9% infertile men and 4.8% of control men. Significant differences in the frequency of the IVS8-5T variant of the CFTR gene have been found between these groups (p = 0.005), as well as between infertile patients without mutations and control men (p = 0.019). In total, the mutations and/or 5T allele have been found in 14.6% of the patients examined. These data indicate increased frequencies of the mutations of the CFTR gene and its allele variant IVS8-5T in Russian infertile men.

[Herpes simplex virus and cytomegalovirus in male ejaculate: herpes simplex virus is more frequently encountered in idiopathic infertility and correlates with the reduction in sperm parameters].

The present investigation was undertaken to study the detection rate of herpes simplex virus (HSV) and cytomegalovirus (CMV) in the ejaculates of males with infertility and to evaluate the impact of virus infection on the major parameters of sperm. Ejaculates from 808 patients were studied. As compared with apparently healthy individuals, the coupled males with primary infertility were found to have HSV more frequently in both the whole ejaculate (31% versus 17%; p = 0.049) and the fraction of actively motile spermatozoa (30% versus 8%; p = 0.016). Ejaculate HSV detection directly correlated with the reduced amount of actively motile spermatozoa (p = 0.0001) and the smaller proportion of morphologically normal forms of germ cells (p = 0.002). CMV was found to have no impact on the motility and morphology of spermatozoids in the ejaculate. Both HSV and CMV in the male ejaculate were significantly more frequently detectable in winter months. The findings lead to the conclusion that HSV is one of the factors for male infertility and can negatively affect the results of assisted reproductive technologies.

[Analysis of germ cell populations in ejaculate of men infected with herpes simplex virus].

Cytological and molecular genetics methods were used to study sperm from patients with sperm infected with herpes simplex virus (HSV) as indicated by virological and immunocytochemical tests. The following methods were used: (1) sperm analysis to evaluate the morphology and functional properties of sperm; (2) fluorescence in situ hybridization (FISH) with DNA probes specific for chromosomes 1, X, and Y to evaluate nondisjunction frequencies of these chromosomes in sperm; and (3) quantitative analysis of immature germ cells in the ejaculate to identify spermatogenic abnormalities. The total sperm count and the count of sperm with normal motility proved similar to the norm. FISH analysis demonstrated no difference in the nondisjunction frequency of chromosomes 1, X, and Y between infertile patients with HSV-infected sperm and fertile donors. Comparative quantitative analysis of immature germ cells from the ejaculate has demonstrated a significant and considerable (threefold) increase in the number of spermatocytes I at the prepachytene stages of prophase I (preleptotene, leptotene, and zygotene) in HSV patients compared to normal donors. At the same time, HSV patients demonstrated a significant decrease in the number of spermatocytes I, a decrease in the proportion of spermatocytes II and spermatids, and a twofold increase in the number of unidentifiable immature germ cells. The data obtained indicate a partial spermatogenic arrest at the early stages of meiotic prophase I in HSV patients, which prompts further research into the cellular mechanisms of abnormal spermatogenesis after viral infection in humans.

[Asymptomatic genital herpes infection and infertility in males].

Incidence of Herpes simplex virus (HSV) was studied in ejaculates of 100 men. The examinees had neither history nor clinical symptoms of HSV genital infection. HSV was detected by a rapid cultural method in the ejaculate of 20 out of 100 examinees (20%). Of 67 males with infertility HSV was detected in 25%, in 19 males examined prophylactically and 14 patients with varicocele (a comparison group) it was found in 10.5 and 7% cases, respectively. Oligozoospermia was two times more frequent in HSV-containing ejaculates than in HSV-negative one. Mean values of the majority of sperm parameters in HSV-positive and HSV-negative groups did not differ statistically. However, it was revealed that such damage of the spermatozoa structure as microhead (consequence of the defective acrosome or reduced genome) and cytoplasm drops on the neck (a sign of immature forms) occurred more often in HSV-infected patients than in persons with HSV-negative ejaculate. Thus, asymptomatic HSV infection has a negative effect on male fertility.

[Herpetic infection of spermatozoa in failure of reproductive technologies and spontaneous abortion].

Herpes simplex virus (HSV) was studied in ejaculate of 36 men whose wives had the history of spontaneous abortions, or failure of artificial insemination (AI), or extracorporeal fertilization (ECF). The control group consisted of 222 patients examined prophylactically after urogenital inflammation or for primary infertility. HSV was detected with immunofluorescence (IF) reaction with monoclonal antibodies to HSV antigens, rapid culture method (RCM) and transmission electronic microscopy. HSV antigens were detected in fraction of mobile spermatozoa in 13 (65%) of 20 samples from patients of the study group. HSV detectability in the study group was significantly higher than in the controls (p = 0.003). Intragametic capsides of HSV were detected at ultrastructural investigation in 13 of 19 (68%) ejaculate samples from patients of the study group in whom IF and/ or RCM results were positive. The study of 6 ejaculate samples of this group in which HSV was not detected by IF or RCM and 24 control samples HSV capsides were not detected. The following conclusions can be made: mobile spermatozoa of normal morphology can be infected with HSV; as shown morphologically, samples of ejaculate from men whose wives had spontaneous abortions contained intragametic HSV capsides; intragametic HSV infection of spermatozoa had a negative effect on the course of pregnancy and/or embryon vitality.

[Effects of sulfate mineral water for drinking in combination with laser and magnetolaser radiation on occurrence of pathological mitoses in spermatogones of irradiated rats and their progeny].

Drinking sulfate mineral water in combination with low-intensive infra-red laser or magneto-laser radiation is shown to effectively depress development of different forms of pathological mitoses in spermatogones of irradiated rats and in their first-generation progeny.

[Types of Y chromosome deletions and their frequency in infertile men].

Deletions of Y chromosome AZF locus were analyzed during a large-scale andrological and genetic examination of 810 infertile men. The search for Yq microdeletions was carried out according to the standard EAA/EMQN guidelines. The breakpoints were mapped for the deletions in AZF locus. The Y chromosome macro- and microdeletions were detected in 61 (7.5%) infertile men. The frequencies of AZF deletions during azoospermia and severe oligozoospermia amounted to 12.2 and 8.1 %, respectively. On the whole, the frequencies of Yq microdeletions and the genophenotypic correlations characteristic of various AZF deletion types comply with the relevant published data. However, spermatozoids in the ejaculate sediment of men with completely deleted AZFa region or AZFb+c deletions (from solitary spermatozoids to several dozens) were detected for the first time. It was demonstrated that the breakpoints were localized between AZFa and AZFb regions proximally to AZFb+c microdeletions for the majority of cytogenetically detectable deletions in the Y chromosome long arm. This indicates that the mechanisms underlying Yq macro- and microdeletions are somewhat different. The issues related to the role of Y chromosome deletions in the origins of monosomy for X chromosome and X/XY mosaicism are discussed.

[Effects of sulfate mineral water and vetoron on the rate of radiation-induced aneuplodia in the bone marrow cells of rats]. / Vliianie pit'evoi sul'fatnoi mineral'noi vody i vetorona na chastotu vozniknoveniia radiatsionno-indutsirovannoi aneuploidii v kletkakh kostnogo mozga krys.

Experiments on non-inbred male rats have found that pretreatment with drinking sulfate mineral water and veteron reduced incidence of radiation-induced aneuplodia in bone marrow cells. The proportion of cells with normal number of chromosomes went up.

[Pattern of hereditary reproductive disorders]. / Struktura nasledstvennykh narushenii reproduktivnoi sistemy.

A comprehensive clinical, endocrinological, and medical genetic studies were made in 678 patients with reproductive disorders and infertility. A schedule was developed to examine the patients with successive current cytological, cytogenetic, molecular cytogenetics and molecular genetic methods. The pattern and proportion of chromosomal and genic abnormalities were defined among the total number of patients with reproductive disorders.

[Spermatogenesis in rats given potable sulfated mineral water in the early postradiation period]. / Sostoianie spermatogeneza u krys pri primenenii pit'evoi sul'fatnoi mineral'noi vody v rannem postradiatsionnom periode.

Rat experiments have shown that course intake (21 days) of sulphate sodium-magnesium-calcium water with sulphate concentration 2.6 mg/l and mineralization 3.1 g/l in early postradiation period (1-2 Gy) inhibits development of dystrophic and destructive processes in spermatogenic epithelium of the testes and maintains spermatogenesis at higher level than control.