[The indicators of anti-endotoxin immunity under preeclampsia].

The dysregulation of immune mechanisms of control of gestational process results in a number of obstetric complications, including preeclampsia. Nowadays, there is no detailed description of characteristics of anti-endotoxin immune response under this complication. To evaluate the role of anti-endotoxin immunity in development of preeclampsia the determination of levels of LBP, IgG to core-region LPS, IgA, IgM, IgG in blood plasma was made in 134 pregnant women during II-III terms. The main group (74 pregnant women with preeclampsia) was divided on 2 sub-groups: 50 women with mild preeclampsia of and 24 women with severe preeclampsia. In the first sub-group, in 14 women with mild preeclampsia additionally concentration of soluble form of CD14 and BPI was determined. The control group consisted of 60 pregnant women with physiological course of gestation. The study demonstrated increase of level of LBP up to 14%, double increase of titers IgG to core-region LPS in blood plasma of pregnant women with preeclampsia under standard concentrations of IgA, IgM, IgG. It was demonstrated that under mild preeclampsia an increase of sCD14, BPI occurs as compared with standard values. The results of study reflect characteristic alterations of congenital and adaptive components of anti-endotoxin immunity under preeclampsia.

[Leading role of infection in formation of placental insufficiency].

AIM: Evaluate the role of endotoxinemia in pregnancy complicated with placental insufficiency. MATERIALS AND METHODS: Complex clinical-laboratory examination of 130 pregnant women in 70 of which placental insufficiency was diagnosed (main group) was carried out. The examinees of the main group were divided into 3 subgroups: 36, 20 and 14 pregnant women with compensated, subcompensated and decompensated placental insufficiency, respectively. The control group was composed of 60 pregnant women with physiological course of gestation. Levels of LPS, LPS-binding protein and IgG against core-region, cytokines (TNF-alpha, IL13, IL2, IL4, IL6, IL8, IL10, IFNgamma), C-reactive protein were determined in blood plasma. Infection by Chlamydia trachomatis, Mycoplasma genitalium, Treponema vaginalis, Neisseria gonorrhoeae, Mycoplasma hominis, Ureaplasma urealyticum, Ureaplasma parvum were evaluated by PCR. RESULTS: In the main group in 64.2% of cases the presence of genital tract infection was established, in 47.0%--urinary system. An increase of LPS level, titers of IgG against LPS core-region and LPS-binding protein in blood plasma of pregnant women with placental insufficiency was shown. Cytokine profile in placental dysfunction was characterized by a significant increase of IL1beta, IL8, TNF-alpha Th1 cytokine and IL10 Th2 cytokine concentrations and a decrease of pro-inflammatory IL2, IFNgamma levels. CONCLUSION: The results indicate a leading role of infection in formation of placental dysfunction as well as prove involvement of LPS Gram-negative bacteria in pathogenesis of this complication.

[Pseudogene PTENP1 5'-region methylation in endometrial cancer and hyperplasias].

Genetic mutations in tumor suppressor gene PTEN are often detected in malignant human cells and these genomic changes are especially characteristic ofendometrial cancer. In our previous researches we assumed that alternative epigenetic mechanism of PTEN inactivation trough promoter methylation may exist in endometrial cancer. Moreover, PTENP1 pseudogene has recently been shown to play a role in positive regulation of PTENgene expression. Taking into account these facts, we analyzed PTEN and PTENP1 methylation status in endometrial hyperplasia and cancer. It was demonstrated that PTENgene promoter was not methylated but PTENP1 was methylated in 11 of 18 endometrial cancers and in 5 of9 endometrial hyperplasias. We can assume that PTENP1 methylation may inhibit transcription of this gene and also PTEN gene transcription trough RNA interference in accordance with ceRNA theory. Thus, aberrant suppression of PTENP1 transcription can play a role in endometrial cancer pathogenesis.

[Unique steroid 21-hydroxylase gene CYP21A2 polymorphism in patients with hyperandrogenism signs].

Hyperandrogenism is a medical condition characterized by excessive production of male sex hormones (androgens) in woman organism. One of the major causes of hyperandrogenism is the autosomal-recessive disorder--congenital adrenal hyperplasia (CAH). The mutational defects in the steroid 21-hydroxylase CYP21A2 gene causing steroid 21-hydroxylase deficiency account for over 90% of CAH cases. Our paper describes the sequencing results of entire CYP21A2 gene from 15 patients with hyperandrogenism signs, which had not nine most prevalent mutations associated with nonclassic CAH as it was previously established. 26 polymorphisms were found by sequencing among which 25 were known previously and 23 of them are referred to "normal" gene variants which do not associated with CAH. At the same time the gene of every patient had unique its own distinctive combination of polymorphisms. New SNP represents synonymous substitution C --> T in 3' part of exon 8. All detected SNPs are not regularly distributed but are clustered along the gene. Notably, they were found in the neighborhood of initiation and termination codons and near the intron-exon boundaries of introns 2, 6 and 8. We hypothesize that "normal" clinically insignificant per se SNPs in unique combinations may influence spatial structure of CYP21A2 mRNA or its pre-mRNA splicing efficiency and decrease gene expression level. This assumption may explain the mechanism of pathological phenotype development in our patients.

[A new DNA diagnostic system for the detection of human CYP21 gene mutations associated with adrenal cortex hyperplasia].

Congenital Adrenal Hyperplasia (CAH) is one of the most widespread severe autosomal recessive hereditary diseases. CAH is caused by the impaired biosynthesis of the key human hormones cortisol and aldosterone and is accompanied by the excess synthesis of androgens. Over 90% of CAH cases are caused by a deficiency of the steroid 21-hydrohylase (P450c21). The degree of damage in this enzyme is responsible for the severity of the clinical manifestation of CAH from potentially lethal to mild symptoms. Various mutations of the gene encoding this enzyme are the main source of the reduced activity of the 21-hydrolase. The location of the highly homological pseudogene CYP21P in close proximity to the functional gene impedes the DNA diagnostics of CAH. To detect the eight most frequent CYP21 gene mutations associated with CAH, we developed a new real-time PCR-based system of DNA diagnostics using new allele-specific primers and TaqMan probes for the analyzed mutations. The method was primarily tested on artificial DNA templates, where the analyzed mutations were introduced by site-directed mutagenesis. Then, it was tested on DNA samples from 43 patients with clinical and biochemical manifestations of CAH; seven patients were used as a control. Two mutant alleles were detected in two different individuals: the nonsense Q318X and the missense V281L mutations.

[Promoters of genes MTHFR from patients with hyperhomocysteinemia and PTEN from patients with malignant and benign endometrial and ovarian tumors].

Mutational changes in the promoter regions of MTHFR genes from patients with hyperhomocysteinemia and PTEN genes from patients with endometrial and ovarian tumors were studied. An increased level of homocysteine was found in a part of the patients with a heterozygous C677T mutation in the MTHFR gene, although a moderate hyperhomocysteinemia is usually associated with homozygous mutation. We hypothesized that, in this case, the allele lacking the C677T mutation may be inactivated by the promoter mutation. The sequencing of both DNA strands of the minimal promoter region of the MTHFR gene in ten patients did not reveal any mutation, which implied another mechanism of the development of hyperhomocysteinemia in these patients. A PCR analysis of the minimal promoter region of the tumor suppressor PTEN in the presence of 2-pyrrolidone in 101 patients from Moscow clinics revealed changes in it in patients with endometrial (56%) or ovarian (29%) cancer, as well as in patients with endometrial hyperplasia and benign ovarian tumors (34.6 and 29%, respectively). It was presumed that the found PTEN gene promoters may arise from epigenetic alterations (erroneous methylation) or may (more rarely) be induced by mutations. As a result of the studies, new molecular markers associated with endometrial and ovarian tumors were revealed and a simple and effective method of detection of these markers was developed.