LABORATORY DIAGNOSTICS OF ACTIVE AND LATENT HHV 6-INFECTION IN PATIENTS WITH HEMATOLOGICAL MALIGNANCIES.

INTRODUCTION: Human herpes virus type 6 (HHV 6) can cause serious infectious complications in immunodeficient patients. It is also capable of integrating into the genome of the infected cell. Due to this, there can be a misdiagnosis between viral integration and active infection during laboratory diagnostics. Thus, determination of HHV 6 infection using proper laboratory tools is relevant. Also the data on viral interference of HHV 6 and other herpes viruses are very poor especially for patients with hematological malignancies. The aim of the study was to identify laboratory markers of HHV 6 and the form of infection in patients with hematological malignancies. MATERIALS AND METHODS: 98 patients with hematological malignancies positive for HHV 6 DNA during the infectious complication were enrolled in the study. Viral load in leukocytes and plasma of peripheral blood, antiviral M and G immunoglobulins and peripheral blood leukocytes count were evaluated. RESULTS: The majority of patients (66 out of 98, 67.3%) showed laboratory signs of latent HHV 6. Integrated HHV 6 was suspected in 2 patients due to high viral load (1.5x105 copies and 1.7x105 copies), but it was not confirmed subsequently. Additional testing of HCMV and EBV in patients with laboratory signs of active HHV 6 infection revealed the superiority of monoinfection over mixed infection (20 of 32, 62.5%). In cases of mixed infection, the most common co-infectant was HCMV observed in 9 out of 12 (75%) cases. Mild leukopenia accompanied HHV 6 active infection. CONCLUSION: Laboratory signs of latent HHV 6 tend to be prevalent in patients with hematological malignancies. In patients with laboratory markers of active HHV 6, the monoinfection demonstrated the superiority over mixed one. In cases of mixed infection, HCMV appeared to be the most commonly co-infectant. No cases of an integrated form of HHV 6 have been observed. The viral load of HHV 6 in leukocytes and blood plasma is almost 3 times lower in patients with a mixed infection than with a monoinfection. Active replication of HHV 6 was accompanied with mild leukopenia.

[Impact of current approaches to laboratory screening of donated blood and its components on hepatitis B virus infection in patients with blood system diseases]. / Vliianie sovremennykh podkhodov k laboratornomu obsledovaniiu donorov krovi i ee komponentov na infitsirovannost' virusom gepatita V u bol'nykh s zabolevaniiami sistemy krovi.

AIM: To evaluate the detection rate of markers for hepatitis B virus (HBV) in the blood samples taken from patients with blood system diseases, by applying the current approaches to examining donated blood and its components for markers of viral infections. MATERIAL AND METHODS: The investigation included blood samples from patients with blood system diseases (n=364) and donors (n=5,011). The results of laboratory screening of donated blood samples (n=13,081) were retrospectively analyzed. Commercial kits of reagents were used for immunochemical assay and polymerase chain reaction. RESULTS: Patients with blood system diseases were recorded to have markers of active HBV infection in 12.6% of cases, anti-HBc in 31.3%, and anti-HBs in 37.6%. A retrospective analysis of the results of screening donated blood samples showed the presence of markers for active HBV infection in 0.28% of cases. A prospective examination of blood donors revealed markers of HBV infection in 4.83% of cases, including those of active forms in 0.54% and anti-HBc in 4.79%. The markers of active HBV infection in donors were only anti-HBc IgM in 0.42% of cases. The blood samples from donors with an anti-HBs titer of >200 mIU/ml contained anti-HBc IgM in 10.5%. CONCLUSION: In the last 5-7 years, the detection rate of markers of HBV infection in the blood samples of patients with blood system diseases have remained at a high level. Screening for decreed markers fails to identify people with inapparent infections among the donors. Even high anti-HBs concentrations in the donated blood may be a risk for HBV transmission by transfusion to a recipient.

[The screening of donor blood on antibodies to nuclear antigen of hepatitis B virus as a tool of increasing of safety of transfusion for patients with diseases of blood system.]

Despite application of decreed modes of laboratory analysis of components of donors' blood, the risk of infection of recipients with hepatitis B virus continues to be actual. The isolated identification of HBsAg provides no control of all categories of persons infected with hepatitis B virus. The analysis of presence of antibodies to nuclear antigen of hepatitis B virus that are the first out of antiviral ones and are preserved for life, is an expedient technique of screening testing of donor's blood that permits implementing an additional selection of donors. During March 2014 - March 2015, cohort of regular anti-hepatitis B virus negative donors of blood and its components. The testing of blood samples for anti-hepatitis B virus can be recommended as a routine test increasing viral safety of blood transfusions for patients with diseases of blood system.

Hepatitis B and C viruses interference in coinfected multitransfused recipients.

Data on hepatitis B (HBV) and c (HCV) viruses interference in hematological patients are described. Patients with a hematological malignancy are at high risk of HBV and HCV infection as recipients of multiple transfusions. Results of the laboratory testing of 339 blood samples of patients treated at the National Research center for Hematology, Russian Federation, were studied. Among these patients, HBV/HCV coinfection markers were observed in 153 patients; HBV markers only, in 76 patients; HCV markers only, in 110 patients. The vast majority of coinfected patients had HBV DNA in blood (significantly more in HBsAg-negative patients: 100% vs. 82.8%, p = 0. 0005). HBsAg-negative coinfected patients had low HBV DNA levels (102-103ME/ml) and reduced (or completely absent) HCV RNA levels. The virus interference leads to a decrease in the viral nucleic acid concentrations. Thus, virus detection should include implementation of high sensitive molecular techniques (such as real-time PCR), and an enhanced set of serological HBV markers along with routine screening methods (HBsAg, anti-HCV).

[RPS4 and NAD5 sequences evidenced of polyphyly of ditrichaceae and parallelisms in the evolution of haplolepidous mosses].

Molecular phylogenetic analysis of 66 representatives of haplolepidous mosses showed polyphylia of Ditrichaceae. According to the data obtained, the structure of the peristome, as well as features of the gametophyte on which a family traditionally allocated, arose independently in different groups of haplolepideous mosses. At least six genera (Distichium, Saelania, Eccremidium, Garckea, Rhamphidium, and Wilsoniella) should be excluded from the Ditrichaceae family, while Saelania and Distichium should be assigned even to another order. The loss of the peristome and forming of cleistocarpous capsules also occurs independently in at least two lineages of Ditrichaceae s. str., and in representatives of several lineages of Pottiaceae, a family derived from this group. Ditrichum, the type genus of Ditrichaceae, is also polyphyletic, species of this genus belong to two clades. It was concluded that parallel lines of the morphological variability in this group of mosses occur and its phylogeny need to be resolved based on molecular data.

Structure of Intergenic Spacer IGS1 of Ribosomal Operon from Schistidium Mosses.

The structure of the intergenic spacer 1 (IGS1) of the ribosomal operon from 12 species of Schistidium mosses was studied. In the IGS1 sequences of these species, three conserved regions and two areas of GC- and A-enriched repeats were identified. All of the studied mosses have a conserved pyrimidine-enriched motif at the 5'-end of IGS1. Species-specific nucleotide substitutions and insertions were found in the conserved areas. The repeated units contain single nucleotide substitutions that make unique the majority of repeated units. The positions of such repeats in IGS1 are species-specific, but their number can vary within the species and among operons of the same specimen. The comparison of IGS1 sequences from the Schistidium species and from representatives of ten other moss genera revealed the presence of common conserved motifs with similar localization. Presumably, these motifs are elements of termination of the pre-rRNA transcription and processing of rRNA.

[The alternative view on diagnostic of hyperlipoproteinemia, cholesterol lipoproteins of low density and effect of statins: a lecture].

The effect of statins occur in several stages: 1) inhibition in hepatocytes of synthesis of functionally specific pool of spirit cholesterol, polar mono-layer of lipoproteins of very low density; 2) activation of hydrolysis of triglycerides in lipoproteins of very low density, formation of apoE/B-100-ligand and absorption of lipoproteins of very low density by insulin-depended cells; 3) decreasing of content of and spirit cholesterol-lipoproteins of very low density in blood plasma; 4) activation of hydrolysis of triglycerides in lipoproteins of low density, formation of apoB-100-ligand and absorption of lipoproteins of low density by insulin-independent cells; 5) decreasing of level of and increasing of content of lipoproteins of high density. During first weeks of effect of statins occurs decreasing of concentration of triglycerides and unesterified spirit cholesterol-lipoproteins of very low density in blood plasma. Then, slower and more durational decreasing of level of spirit cholesterol-lipoproteins of low density occurs. The value of spirit cholesterol-lipoproteins of low density is primarily determined by content of palmitic saturated fatty acid in food, its endogenous synthesis from glucose and concentration of palmitic triglycerides and lipoproteins of very low density of the same name in blood plasma. The effect of preparations is biologically valid and corresponds to alternative hypolipidemic preparations. All these preparations have an effect following a common algorithm: they activate, using different mechanisms, receptor absorption of lipoproteins of very low density or lipoproteins of low density by cells. The level of spirit cholesterol-lipoproteins of low density in full measure depends on content of triglycerides in blood. The concentration of spirit cholesterol in blood plasma has a reliable diagnostic significance only under physiological content of triglycerides. The main criterion of diagnostic and control of hypolipidemic therapy biologically is content of triglycerides. The comprehension of differences in effect of hypolipidemic preparations within framework of common algorithm permits rationally combine them under treatment of both primary inheritable phenotypes of glucolipoproteins and secondary symptomatic types of glucolipoproteins under obligatory observation of strict dietary treatment.

[The PCR markers of viral infections under chronic gastritis in children].

The extended monitoring (up to 1 year 11 months) of PCR markers was implemented concerning viral infections: cytomegalovirus, Epstein-Barr virus, simple herpes virus type I and II, hepatitis B virus, hepatitis C virus and bacterial infection of Helicobacter pylori in bioassays (blood, biopsy material of mucous coat of stomach and inferior third of esophagus) from children with different types of chronic gastritis. In biological samples from patients with gastritis type A and type A + B DNA of hepatitis B virus (87% and 71% of patients correspondingly) and DNA of Epstein-Barr virus (63% and 67% of patients) were detected with high rate. Under gastritis type B and C these markers were detected significantly rarely (20-36%). Among patients with gastritis type A, B and A + B, the positive results on DNA of cytomegalovirus consisted 13-17%. In patients with gastritis type C DNA of cytomegalovirus was not detected. In any of analyzed samples no DNA of simple herpes virus type I and II was detected. The control of DNA of H. pylori demonstrated its presence in biological materials of 67% and 84% of patients with gastritis type B and A +B. This type of DNA was absent in patients with gastritis type A and C. Under gastritis type A, B and A+B, DNA of Epstein-Barr virus and DNA of hepatitis B virus detected more often in biological materials of mucous coat of stomach (71%-100%) and out of them simultaneously in blood in 33%-60% of examined patients and only in blood up to 29%. DNA of Epstein-Barr virus was detected in leukocytes of peripheral blood and DNA of hepatitis B virus both in plasma and leukocytes of peripheral blood. Under gastritis type C DNA of Epstein-Barr virus was always detected in leukocytes of peripheral blood (in 20% out of these patients simultaneously in biological material) and DNA of hepatitis B virus just as much in blood (plasma and/or leukocytes of peripheral blood) and biological materials. The lower concentrations (less than 700 copies/ml) DNA of hepatitis B virus in most samples were detected in absence of markers of hepatitis B virus. In patients with autoimmune gastritis and in absence of bacterial infection H. pylori (group I) or against its background (group III) PCR-markers of hepatitis B virus and Epstein-Barr virus were detected quite often. The evidence of persistence (in superior sections of digestive organs) of Epstein-Barr virus nad hepatitis B virus is detection of DNA of these viruses under their extended monitoring (up to 1 year 11 months) in biological samples from patients with autoimmune forms of gastritis type A and type A+B.

Analysis of circulating tumor cells in patients with triple negative breast cancer during preoperative chemotherapy.

The presence of circulating tumor cells in the blood of patients with triple negative breast cancer (early and locally advanced cancer) before and after preoperative chemotherapy was assessed using expression markers. Before therapy, circulating tumor cells were detected in 5 of 13 (38%) patients with early cancer and in 7 of 17 (41.2%) patients with locally advanced cancer. After therapy, the circulating immune cells were detected in one patient with locally advanced cancer, who had no circulating cells before therapy. The tumor was resistant to chemotherapy and the disease progressed. The detected circulating tumor cells were HER-2-positive, while the primary tumor was HER-2-negative. It was concluded that the circulating immune cells can be a potential marker of the efficiency of therapy and predictors of the disease course, while their phenotype can differ from the phenotype of the primary tumor.

[Genotyping of BRCA1, BRCA2 and CHEK2 germline mutations in Russian breast cancer patients using diagnostic biochips].

Germline mutations of BRCA1/2 genes cause the predisposition of their carriers to breast or/and ovary cancers (BC or/and OC) during the lifetime. Identification of these mutations is a basis of molecular diagnosis for BC susceptibility. Rapid genotyping technique using microarrays for identification of BRCA1 185delAG, 300T>G, 4153delA, 5382insC mutations and 4158 A>G sequence variant; BRCA2 695insT and 6174delT mutations; 1100delC mutation in CHEK2 gene was applied for 412 randomly collected breast cancer samples from the central region of European area of Russia. In 25 (6.0%) patients (6.0%) BC was associated with other tumours: OC, cervical cancer, colorectal cancer etc. BRCA1/2 and CHEK2 mutations were found in 33 (8.0%) BC patients. The most frequent mutation was BRCA1 5382insC, occurred in 16 (3.9%) BC patients, and CHEK2 1100delC, revealed in 7 (1.7%) BC patients. An application of diagnostic BC-microarray for genetic testing of BRCA1/2 and CHEK2 founder mutations has been discussed.

[Discordance of estrogen receptor status between primary and metastatic breast cancer--possible reasons and prognostic value].

The aim of the review was systematization of the data on discordance in expression of estrogen receptors between primary and metastatic breast cancer, different metastases and repeated analyses of the same tissue. The possible reasons for the phenomenon are discussed. The authors emphasize the need to analyze estrogen receptors in breast cancer metastases, regardless of the receptor status of the primary tumor, for predicting the course of the metastatic disease and providing an adequate treatment of the metastatic tumor in strict accordance with its receptor status during drug therapy. The works cited in the search engine Pub Med to May 2013 were analyzed.

[Comparative analysis of occupational risk evaluation results through various methodic approaches].

Comparative analysis covered three models of occupational risk evaluation: (1) Method to evaluate risks at workplace, elaborated in Finland and recommended by International Work Safety Agency for Eastern Europe and Central Asia countries; (2) Method elaborated in Research Institute for Occupational Medicine with RAMSc under the direction of N.F. Izmerov and E.I. Denisov; (3) Method evaluating individual occupational risk, with consideration of work conditions and worker's health state, also elaborated in Research Institute for Occupational Medicine with RAMSc and Klin Institute of Work conditions and safety, approved in 2011 as methodic recommendations. Findings are that in contemporary Russia a unified method evaluating occupational risk is more expedient; the first method satisfactorily describes actual ratio of occupational risk levels and could be useful as an additional method for its evaluation, especially for psycho-social factors; the second method does not allow to evaluate occupational risk acceptably if absent longstanding occupational morbidity, but is recommended for risk evaluation in evidence-based medicine; the third method is recommended at federal level to fulfil requirements of Labour Code in Russian Federation and obligatory social insurance purposes.

[The phylogeny of Schistidium (Bryophyta, Grimmiaceae) based on primary and secondary structure study of nuclear rDNA internal transcribed spacers].

The phylogeny of Schistidium (Bryophyta, Grimmiaceae) was studied on the basis of nucleotide sequences of internal transcribed spacers ITS1-2 of nuclear DNA and trnT-trnD region of chloroplast DNA. The consistency of phylogenetic trees constructed from nuclear and chloroplast sequences was shown. A basal grade and two large clades were resolved on the phylogenetic trees. Morphological characteristics specific for these clades were described. ITS1 and ITS2 secondary structures of Schistidium species were modeled using thermodynamic criteria. Four different structures of the longest ITS1 hairpin were identified. Possible paths of Schistidium evolution were considered based on the four types of ITS1 secondary structure and phylogenetic trees.

[Laboratory diagnosis of herpesvirus infections in patients with acute leukemias].

The paper presents the results of monitoring the markers of herpes simplex viruses types 1 and 2, cytomegalovirus, Epstein-Barr virus, and human herpesvirus type 6 in the blood and bone marrow of patients with acute leukemias during induction multidrug therapy. Whether it is expedient to diagnose herpesvirus markers in patients with acute leukemias in the period of remission induction is discussed.

Stimulatory effect of green tea extract on the growth of neurites in the rat spinal ganglion culture.

Addition of green tea extract in concentrations of 0.004-0.006% to the nutrient medium markedly stimulated the growth of spinal ganglion neurites of 1-2-day-old rats.

Baikal skullcap extract stimulates neurite growth in cultures of rat dorsal root Ganglia.

The effect of baikal skullcap extract on the development of dorsal root ganglia from 1-2-day-old rats in organotypic cultures was studied. Baikal skullcap extract produced a dose-dependent stimulating effect on neurite growth in neurons of dorsal root ganglia.

[Search for lectin producers among the yeast of Pichia Hansen emend. Kurtzman genus]. / Poisk produtsentov lektinov sredi drozhzhei roda Pichia Hansen emend. Kurtzman.

Strains (103) of yeast from Pichia genus: P. anomala, P. guilliermondii and P. membranaefaciens species have been investigated. The researchers have found 87 strains (84.5%) capable to form extracellular and surface lectins with activity to 512 hemagglutinating units. The overwhelming number of strains (75.7-92.6%) synthesized lectins of the both forms. Considerably less quantity of lectin producers of some form--extracellular or surface ones--have been found. Availability of producers of extracellular lectins was characteristic to greater extent of the yeast of P. anomala and P. guilliermondii, and producers of surface lectins--mainly of P. guilliermondii. The most active producers of lectins have been isolated from human and animal organisms, soils and wine.

[On the identity of Candida utilis and Pichia jadinii yeast species]. / Ob identichnosti vidov drozhzhei Candida utilis i Pichia jadinii.

Life cycles of Candida utilis (Henneberg) Lodder et Kreger-van Rij imperfect yeast (13 strains) used in industry were the study subject. When the strains were mated, we detected one of the stages of sexual process--conjugation of cells of the opposite mating type. Most of the studied cultures conjugated on the 2d-3d day. No ascospores were formed. Haploidy and heterothallism of the studied C. utilis strains were confirmed by hybridization of auxotrophic mutants. Based on PCR assay results, the yeasts are demonstrated to belong to the ascospore perfect yeast species of Pichia jadinii (A. et R. Sartory. We ill et Meyer, Wickerham) Kurtzman.

Development and maintenance of otoconia: biochemical considerations.

The first part of this review deals with recent advances in the understanding of biochemical mechanisms of otoconial morphogenesis. Most important in this regard is the molecular characterization of otoconin 90, the principal matrix protein of mammalian calcitic otoconia, which was found to be a homologue of the phospholytic enzyme PLA2. The unique and unexpected expression pattern of this protein required radical rethinking of traditional concepts. The new data, when integrated with existing information, provide a rational basis for an explanation of the mechanisms leading to crystal nucleation and growth. Based on this information, a hypothetical model is presented that posits interaction of otoconin 90 with microvesicles derived from the supporting cells as a key event in the formation of otoconia. The second part of the review is directed at the controversial subject of maintenance of mature otoconia and systematically analyzes the available indirect information on this topic. A synthesis of these theoretical considerations is viewed in relation to the pathogenesis of the important otoneurologic entities of BPPN and senile otoconial degeneration. The last part of the review deals with several animal models that promise to help elucidate normal and abnormal mechanisms of otoconial morphogenesis, including mineral deficiencies, mutations with selective otoconial agenesis, as well as targeted disruption of essential genes.

[Determination of type-specific synthetic peptide antibodies to hepatitis delta virus ]. / Opredelenie tipospetsificheskikh antitel k virusu gepatita del'ta na osnove sinteticheskikh peptidov.

Nine peptides from immunodominant (53-68 and 65-80 aa) and hypervariable (201-213 aa) regions derived from delta antigen sequences corresponding to 3 HDV genotypes were synthesized. Type specificity of antibodies to the resultant peptides was evaluated by indirect enzyme immunoassay with sera from patients with hepatitis D and asymptomatic carriers of anti-delta antibodies. Analysis of the results showed that HDV circulating in the Central Volga region belongs to type I in the majority of cases. High heterogeneity and changeability of HDV genome during the period of transition from acute forms to asymptomatic carriership was revealed. Possibility of circulation of new hybrid HDV strains causing mixed infection in Russia is discussed.