Alexander M. Gzgzyan

Genome-wide 5-hydroxymethylcytosine patterns in human spermatogenesis are associated with semen quality

We performed immunofluorescent analysis of DNA hydroxymethylation and methylation in human testicular spermatogenic cells from azoospermic patients and ejaculated spermatozoa from sperm donors and patients from infertile couples. In contrast to methylation which was present throughout spermatogenesis, hydroxymethylation was either high or almost undetectable in both spermatogenic cells and ejaculated spermatozoa. On testicular cytogenetic preparations, 5-hydroxymethylcytosine was undetectable in mitotic and meiotic chromosomes, and was present exclusively in interphase spermatogonia Ad and in a minor spermatid population. The proportions of hydroxymethylated and non-hydroxymethylated diploid and haploid nuclei were similar among samples, suggesting that the observed alterations of 5-hydroxymethylcytosine patterns in differentiating spermatogenic cells are programmed. In ejaculates, a few spermatozoa had high 5-hydroxymethylcytosine level, while in the other ones hydroxymethylation was almost undetectable. The percentage of highly hydroxymethylated (5-hydroxymethylcytosine-positive) spermatozoa varied strongly among individuals. In patients from infertile couples, it was higher than in sperm donors (P<0.0001) and varied in a wider range: 0.12-21.24% versus 0.02-0.46%. The percentage of highly hydroxymethylated spermatozoa correlated strongly negatively with the indicators of good semen quality - normal morphology (r=-0.567, P<0.0001) and normal head morphology (r=-0.609, P<0.0001) - and strongly positively with the indicator of poor semen quality: sperm DNA fragmentation (r=0.46, P=0.001). Thus, the immunocytochemically detected increase of 5hmC in individual spermatozoa is associated with infertility in a couple and with deterioration of sperm parameters. We hypothesize that this increase is not programmed, but represents an induced abnormality and, therefore, it can be potentially used as a novel indicator of semen quality.We performed immunofluorescent analysis of DNA hydroxymethylation and methylation in human testicular spermatogenic cells from azoospermic patients and ejaculated spermatozoa from sperm donors and patients from infertile couples. In contrast to methylation which was present throughout spermatogenesis, hydroxymethylation was either high or almost undetectable in both spermatogenic cells and ejaculated spermatozoa. On testicular cytogenetic preparations, 5-hydroxymethylcytosine was undetectable in mitotic and meiotic chromosomes, and was present exclusively in interphase spermatogonia Ad and in a minor spermatid population. The proportions of hydroxymethylated and non-hydroxymethylated diploid and haploid nuclei were similar among samples, suggesting that the observed alterations of 5-hydroxymethylcytosine patterns in differentiating spermatogenic cells are programmed. In ejaculates, a few spermatozoa had high 5-hydroxymethylcytosine level, while in the other ones hydroxymethylation was almost undetectable. The percentage of highly hydroxymethylated (5-hydroxymethylcytosine-positive) spermatozoa varied strongly among individuals. In patients from infertile couples, it was higher than in sperm donors (P<0.0001) and varied in a wider range: 0.12-21.24% versus 0.02-0.46%. The percentage of highly hydroxymethylated spermatozoa correlated strongly negatively with the indicators of good semen quality – normal morphology (r=-0.567, P<0.0001) and normal head morphology (r=-0.609, P<0.0001) – and strongly positively with the indicator of poor semen quality: sperm DNA fragmentation (r=0.46, P=0.001). Thus, the immunocytochemically detected increase of 5hmC in individual spermatozoa is associated with infertility in a couple and with deterioration of sperm parameters. We hypothesize that this increase is not programmed, but represents an induced abnormality and, therefore, it can be potentially used as a novel indicator of semen quality.

Genomic distribution of 5-formylcytosine and 5-carboxylcytosine in human preimplantation embryos: PENDINA et al.

5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) are derivatives of 5-methylcytosine (5mC). Sequential enzymatic oxidation of 5mC produces 5-hydroxymethylcytosine (5hmC), 5fC, and 5caC. Similarly to 5hmC, 5fC and 5caC serve as transient intermediates in active DNA demethylation which is a crucial step in epigenetic reprogramming of mammalian preimplantation embryos. This article is protected by copyright. All rights reserved.

Pathogenetic justification of possibility of antidiuretic hormone use in incontinent women

Hypothesis/aims of study. Urine incontinence seems to include several pathogenetic forms, as efficient therapy is provided by different medications. Commonly used in the treatment of female patients with overactive bladder and nocturnal polyuria is desmopressin which normalizes the water excretion of the kidney, which is disturbed by a presumed inverted rhythm of vasopressin secretion in these patients. The current analysis was undertaken to evaluate the clinical efficiency of desmopressine in incontinent patients with nocturnal polyuria and polyuria. Study design, materials and methods . A total of 84 patients with complaints of urinary incontinence, polyuria (24-urine volume of 40 mL/kg bodyweight or above) or nocturnal polyuria (nocturnal volume/24-h urine volume of 0.33 or above) and 14 control subjects were included. Mean patient age was 43.6 ± 4.6 years, in control subjects 38.5 ± 6.4 (p > 0.05). All participants performed 24h-urinecollection to determine the voided volumes and the levels of creatinine, osmolality, sodium, magnesium and potassium for each sample. A blood sample was taken during the 24-urinecollection to determine the levels of creatinine, osmolality, sodium, magnesium and potassium. The examination of patients with polyuria and nocturnal polyuria was performed twice: in the initial state and one month after the start of treatment with optimal dose of desmopressin. Optimal dose was established through an open-label dose-titration using 0.1 mg, 0.2 mg and 0.4 mg of desmopressin (Minirin). Safety parameters assessed included incidence of adverse events, vital signs and serum sodium levels. Results . In patients with polyuria and nocturnal polyuria the glomerular filtration rate was normal, whereas diuresis and solute (sodium, magnesium, potassium) excretion in night samples in nocturnal polyuria and both in night and day samples in polyuria were increased. The higher diuresis and the higher solute excretion observed in nocturnal polyuria and polyuria are accompanied by an increase of free water reabsorption. In nocturnal polyuria and polyuria a high correlation was found between the free water reabsorption and solute excretion. This occurs against the background of the high night and day osmotic concentration. The statistically significant recovery of renal function occurred in 12 incontinent women with polyuria and 18 with nocturnal polyuria. In these papients there was a statistically significant decrease in diuresis, osmolar clearance and excretion of sodium, potassium and magnesium. Concluding message . As desmopressin affects cells of the thick ascending limb of Henle’s loop, it is likely that nocturnal polyuria and polyuria result from a disturbed regulation of the function of these cells. Normalization can be achieved by desmopressin administration to stimulate V2-receptors, which increase water permeability and water reabsoption in collecting ducts as well as ion reabsorption by cells of the thick ascending limb of Henle’s loop.

Renal ionoregulation function in incontinent women

Renal ionoregulation function was studied in 143 women with stress incontinence, in 43 with urge incontinence and in 91 with mixed incontinence. Total polyuria was diagnosed in 8,7 ± 1,7% and nocturnal polyuria in 21,7 ± 2,5% incontinent wonen. Changes in kidney function in total and nocturnal polyuria appear to be due not to a decrease in water reabsorption in the renal collecting duct but to reduction of ion reabsorption in the thick ascending limb of the Henle loop. Due to this defect, reabsorption of ions and water is decreased; as a result, lager volumes of fluid enter the collecting ducts.

Endometrial stem cells expansion capability for local and systemic routes of administration in a model of experimentally injured endometrium

Endometrial stem cells due to their therapeutic characteristics could to be an effective tool of cell technologies in reproductive medicine. The aim of the study was to determine the most therapeutically effective route of administration for endometrial stem cells suspension. The study was conducted in approved animal model of injured endometrium. To create the experimental model tissue pieces of autologous endometrium were implanted on the anterior abdominal wall peritoneum using general surgical techniques. Experimental group animals were treated with endometrial stem cells suspension; in the control animal group a placebo was used. Local and systemic routes of endometrial stem cells administration were compared. The direct injections of stem cells suspension in the endometrial implants were used as the local route of administration, the intravenous injections of stem cells suspension were used as a systemic route. Endometrial stem cells expansion didn’t depend on the routes of administration, whereas therapeutic effects of stem cells was more obvious in tissue pieces after local injection of stem cells.

Effectiveness of growth hormone use in women with “poor” ovarian response to the stimulation of gonadotropins in IVF/IVF+ICSI programs

The use of growth hormone in women “poor” response groups to the ovulatory stimulation in IVF cycles raises ovarian sensitivity to the gonadotropin exogenous influence, this way, increasing the probability of pregnancy. Taking into consideration small sample of women examined, it is necessary to continue the study of growth hormone use effectiveness and safety for the recommendation of its wide administration in clinical practice.

Expected applications of the endometrial stem cells in clinical practice

Recent findings in stem cell biology and cell engineering cannot be left without physicians’ attention. Endometrium as actively regenerating tissue represents also a source of endometrial stem cells. Endometrial stem cells have specific properties that distinguish them from other somatic stem cells. Endometrial stem cells have already shown their therapeutic efficiency and safety in a number of experimental and clinical trials. The biological basis of endometrial stem cells therapeutic potential and recent data on their use in clinical practice are discussed in the article.

Role of kidney function change in development of low urinary tract during pregnancy

Background. During pregnancy many women suffer from lower urinary tract symptoms (LUTS). The etiology and pathogenesis of urinary loss during pregnancy remain still unclear. Normal pregnancy is known to be associated with functional changes in various physiological systems. Changes of kidney function, on the one hand, are the part of physiological changes during pregnancy, on the other hand - can promote urinary incontinence. Purposes. The purpose of this study was to estimate the role of kidney function changes in development of LUTS during pregnancy. Materials and methods. A total of 270 women with the physiological course of pregnancy (83 in the I first trimest, 78 - in the II trimester and 109 - in the III trimester) aged from 18 till 44 years, with no LUTS before the current pregnancy were examined. Blood serum and urine osmolality was determined with MT-4 milliosmometer (“Burevestnik”, Russia); the concentration of serum and urine creatinine, sodium, potassium and magnesium ions were determined with automatic Abbott Architect 8000 analyzer. LUTS are revealed in 81.9 ± 2.3% of pregnant women. Urinary loss in pregnant women is combined with increase in diuresis. Grater diuresis is due to increasing osmotically active substances excreation, precisioly sodium. Rise in nocturnal diuresis does not lead to inversion of circadian rhythm of urine production. A direct correlation has been found between urine output and glomerular filtration rate.

Diskussionnye voprosy vedeniya zhenshchin s antifosfolipidnymi antitelami pri primenenii VRT

There are many controversies in the management antiphospholipid antibodies-positive women undergoing assisted reproductive technologies. Controversial and important is not only the effect of antiphospholipid antibodies (APA) on the success of IVF, but also approaches to the management of this group of women. The aim of this work was to study the prevalence of antiphospholipid antibodies in women with 3 or more IVF failures and effectiveness of IVF in women with APA treatment with membrane plasmapheresis before IVF and intravenous immunoglobulin during IVF. In women with 3 or more IVF implantation failure APA were detected in 35.95 % of cases. The use of intravenous immunoglobulin along with the standard therapy is a safe and effective for improvement of IVF outcomes in women with antiphospholipid syndrome (APS) or the presence of APA. The most effective is a combination of the membrane plasmapheresis therapy before IVF and intravenous immunoglobulin during IVF standard therapy including LMWH and low-dose aspirin in IVF protocol. This therapy in patients with the presence of APA increases the pregnancy rate after IVF to 46.15 %.

Doppler indices of uterine vessels in assessing the ability of endometrial implantation during in vitro fertilization (IVF) cycles

Embryo implantation is a multifactorial process. IVF success depends on the quality of the embryo and the receptivity of the endometrium. Ultrasound is non-invasive method to assess the answer of the ovaries and the endometrium in IVF cycles. Color Doppler mapping of the uterine vessels provides information about the degree of vascularization of the uterus and endometrium and ready for implantation. In this article are represented characters uterine blood vessels of 28 infertile women undergoing IVF embryo transfer, followed by 2–3 days of the menstrual cycle, on the day of the trigger and the day of embryo transfer. The comparison of hemodynamic parameters (systolic-diastolic ratio S/D, index of resistance IR, pulsation index PI) of uterine vessels (uterine, arcuate, radial, basal and spiral arteries) was made in 15 pregnant and 13 non-pregnant women after IVF treatment. It was found, that the hemodynamic parameter of uterine vessels on 2–3 days of menstrual cycle does not reflect the ability of endometrium to implantation in IVF cycles. The most effective in the forecast of pregnancy was to determine the mentioned hemodynamic parameters in the basal, spiral arteries of the uterus on the day of the trigger and basal IR and spiral PI arteries on the day of embryo transfer.