[Competitive ELISA test system for the detection of antibodies to the rabies virus in animals (Rhabdoviridae: Lyssavirus)].

INTRODUCTION: The main approach to the rabies prevention is the vaccination of domestic and wild carnivores. For the routine evaluation the anti-rabies vaccination effectiveness, World Organization for Animal Health (OIE) recommends various enzyme-linked immunosorbent assays (ELISA).The aim of the study was to design and validate a competitive ELISA (cELISA) test system for the detection of antibodies to the rabies virus (RABV). MATERIALS AND METHODS: The development of the cELISA was carried out following the OIE recommendations. RESULTS: The repeatability of the cELISA results within one laboratory was satisfactory (coefficient of variation 7.95-13.61%). The coefficient of determination (CD) between the results of the virus neutralization reaction (FAVN) and cELISA was 0.988, p < 0.001. The lower threshold for antibody detection was less than 0.02 IU/ml. The cELISA did not demonstrate cross-reactivity against antibodies to canine distemper virus, parainfluenza virus, parvovirus, coronavirus, and canine adenovirus (types I and II). During the study of 137 dog blood sera, diagnostic specificity (DSp) and diagnostic sensitivity (DSe) for the cELISA were 83.1% and 94.9%, respectively, and CD between the cELISA and FAVN results was 0.968, p < 0.001. DISCUSSION: Indirect ELISA test systems for determining the level of antibodies to RABV are not sensitive enough compared to reference tests, unlike cELISA. The developed test system is not inferior for its DSp and DSe to the commercial cELISA BioPro ELISA Rabies Ab (DSp 66.7%, DSe 94.4%). CONCLUSION: The developed cELISA test system can be used to detect antibodies to RABV in the blood serum of dogs for evaluating the effectiveness of mass vaccination programs.

[Vitamin B12 is a diagnostic marker of decreased men reproductive function]. / Vitamin B12 kak diagnosticheskii marker snizheniia reproduktivnoi funktsii u muzhchin.

Although the relationship between the amount of vitamin B12 and the quality of sperm exists, but results are controversial and require several additional research. The objective of our study was to analyse the amount of vitamin B12 in the sperm samples from patients with chronic prostatitis and varicocele with accompanying asthenozoospermia, and to identify the relationship between the amount of vitamin B12 and asthenozoospermia. The research was carried out of men with chronic prostatitis and varicocele with accompanying asthenozoospermia and infertility at the age of 27±2 years. The material of the investigation was spermoplasm. A chemical microscopic examination of the ejaculate was carried out with a sperm analyzer and with the light microscopy. The amount of vitamin B12 in the spermoplasm was determined by the method of competitive ELISA. It was found that the level of vitamin B12 was 3.6 times lower in patients with chronic prostatitis III B and asthenozoospermia then in the control group. Among patients with varicocele of II and III grade and asthenozoospermia, the level of vitamin B12 was 4.4 times lower than in control group. A positive correlation relationship of average strength was revealed (r=0,683; p=0,001). Additionally, it was revealed that among patients with Chronic prostatitis III B and varicocele of II and III grades with accompanying asthenozoospermia, there was the positive correlation relationship of average strength (r=0,690; p=0,001) between the amount of vitamin B12 and sperm mobility. A decrease in vitamin B12 levels may serve as a marker of reproductive dysfunction in men.

Spherical particles derived from TMV virions enhance the protective properties of the rabies vaccine.

In this study the ability of spherical particles (SPs) obtained from the tobacco mosaic virus (TMV) virions to enhance the immunogenic potential of the vaccine was evaluated. TMV SPs were shown to increase the protective properties of the widely used effective Russian adjuvant-free rabies vaccine, composed of killed rabies virions. The results of the NIH potency test showed enhancement of protectivity, that is comparable with the effect of the incomplete Freund׳s adjuvant on the same vaccine.

[The main metabolites ferrokinetics in differential diagnostic of anemic syndrome with microcytic, hypochromic characteristics of erythrocytes in patients with colorectal cancer.]

The clinical analysis of blood and main metabolites of ferrokinetics was implemented before treatment in 46 patients with colorectal cancer mainly in stage III-IV. The analysis of obtained results demonstrated that the technique of detection of hepcidin 25 can be used as an additional diagnostic criterion of anemic syndrome in complex with ferritin, soluble receptors of transferrin and expanded clinical analysis of blood. This is especially important in differential diagnostic of anemia of chronic diseas combined with iron-deficiency anemia and anemia of chronic disease with functional iron deficiency because treatment differs in the main. The perspective consists in pathogenic approach to treatment of anemia of chronic disease in oncologic patients using anti-hepcidin medications on the basis of antibodies to hormone, inhibitors and blockers of its expression under control of concentration of hepcidin.

[The disorders of hemostasis system and thrombotic complications in oncologic patients having liver operation.]

The surgical interventions in patients with malignant tumors of liver cause a development of various forms of syndrome of disseminated intravascular coagulation (DIC) with significant increasing of concentration of markers of intravascular blood coagulation against the background of dramatic decreasing of activity of factors of prothrombin complex, level of natural anti-coagulants and components of fibrinolytic system. The thrombosis of veins of various localization developed in 12 patients with subacute form of DIC. The application of low-molecular heparin in combination with concentrate of anti-thrombin III and freshly frozen plasma decreases intensity of intra-vascular blood coagulation and results in reduction of factors of coagulation and is an efficient and adequate method of treatment of thrombosis and thromboembolism of pulmonary artery in oncologic patients. The significant increasing of level of D-dimer, von Willebrand factor against the background of dramatic decreasing of prothrombin activity, content of antithrombin III and level of plasminogen are hemostasiologic factors of development of severe liver insufficiency. The corresponding timely diagnostic and correction permits to ameliorate results of surgical treatment of patients with malignant tumors of liver.

[Radionecrosis after radiation synovectomy]. / Radionekrose nach Radiosynoviorthese.

Intra-articular radiation synovectomy is an established and well documented therapy in rheumatology. A rare side effect is necrosis of surrounding tissue caused by leakage of the radionuclide.We describe the case of a 34-year old patient with rheumatoid arthritis who developed skin necrosis 4 weeks after an intra-articular radiation synovectomy of the left wrist. When these lesions are recognized early surgical therapy or hyperbaric oxygen therapy can be initiated.

[THE DIFFERENTIAL DIAGNOSTIC POSSIBILITIES IN EVALUATION OF IRON-DEFICIENT CONDITION UNDER ANEMIAS].

The article presents data concerning differential diagnostic possibilities of evaluation of genuine iron-deficient anemia and anemia of chronic diseases. The variety of mechanisms of development of anemia of chronic diseases is demonstrated, including effect of humoral inhibitors of erythropoiesis, disorder of iron metabolism at the expense of its redistribution into cells of macrophage system, suppression of erythropoiesis resulted in redistributed or functional iron deficiency. The data is presented concerning significance in diagnostic of anemia of chronic diseases of such factors as content of ferritin, dissolving receptors of transferrin and role of hepcidin protein in pathogenesis of anemia of chronic diseases. The analysis of scientific publications demonstrated that hepcidin is a negative regulator of iron metabolism. Under iron-deficient anemia its level in blood decreases that contribute to extensive absorption of iron in gastrointestinal tract. On the contrary, under anemia of chronic diseases its content drastically increases and results in blocking of iron transport everywhere, including internal epithelium, macrophages, placenta and other types of cells. The hyper-production of hepcidin during infection and inflammation is responsible for anemia of chronic diseases. The perspectives of development of pharmaceuticals decreasing level of hepcidin for treatment of anemia of chronic diseases is demonstrated.

[Differential diagnosis of anemia in true and functional iron deficiency in patients with chronic diseases (malignant tumors)]. / Differentsial'naya diagnostika anemicheskogo sindroma pri istinnom zhelezodefitsitnom sostoyanii i funktsional'nom defitsite zheleza u bol'nykh s khronicheskimi zabolevaniyami (zlokachestvennymi novoobrazovaniyami).

AIM: To make a differential diagnosis of true and functional iron deficiency (ID) in anemia of chronic diseases (ACD). MATERIALS AND METHODS: A total of 540 patients (785 investigations) and 311 healthy individuals (a control group) were examined. Among them, there were 118 cancer patients admitted for surgical treatment and 226 for chemotherapy (CT), 56 blood cancer patients, 86 with breast diseases without anemia, and 54 workers of the institute with the signs of ID. An enzyme immunoassay was used to determine the plasma levels of ferritin (FR), soluble transferrin receptor (sTFR), and endogenous erythropoietin (EPO). RESULTS: 183 cancer patients did not receive specific treatment, 161 had different cycles of CT. In true ID, anemia was microcytic (average red blood cell size, <80.0 fl) and hypochromic (mean corpuscular hemoglobin concentration, <27.0 pg) with low FR levels (less than 20 ng/ml) and high sTFR concentrations (more than 4.0 µg/ml). The EPO level corresponded to the degree of anemia and was 4-5-fold the normal values. In ACDs, 292 (55%) patients had normocytic normochromic anemia with high FR levels (more than 100 ng/ml), the level of sTFR was low and that of EPO did not correspond to the degree of anemia in 74% of the patients. Functional ID was detected in 108 (37%) cancer patients in the presence of ACD. Despite high FR levels and low sTFR, anemia in Hodgkin's and non-Hodgkin's lymphomas was hypochromic microcytic, which may be considered as severe functional ID in ACD. CONCLUSION: In addition to a clinical blood test, the pharmacokinetic parameters (FR, sRFR, and EPO) should be determined for the differential diagnosis of true and functional ID.

[The main metabolites of ferrokinetics in differentiated diagnostic of anemic syndrome].

The article presents data concerning clinical significance of interleukin 6, hepcidin-25 and its predecessor prohepcidin in differentiated diagnostic of anemic syndrome. The sampling of 192 workers of the oncologic center undergoing annual dispensarization was examined. The genuine reference values for immune enzyme (interleukin 6, hepcidin-25, prohepcidin, soluble receptors of transferrin, ferritin, erythropoietin) techniques are presented. The analysis of results demonstrated that values of interleukin 6, hepcidin-25, prohepcidin can be applied as additional diagnostic criteria of anemic syndrome and especially in differentiated diagnostic of anemia of chronic disease with functional iron deficiency. The carried out study is a new perspective direction of modern concept of iron metabolism in patients with anemic syndrome.

[Methodological aspects of the assessment of phytotoxicic properties of ice-melter reagents].

One of the main criteria which determine the possibility of the use of a particular type of ice-melter reagents (IMR) is the degree of their safety for the environment and human health, which is reflected in the establishment of safe doses and concentrations. In this regard, the current area of research is to improve the ecological and epidemiological principles of risk assessment of modern types of anti-icing agents. Currently available data concerning monitoring soil studies and the snow held in various cities of Russia, show that there is a process of accumulation of the main components of IMR - sodium and chlorine ions in the areas related to the roadway. The article is designated a problem of existing methodological approaches to the assessment of the phytotoxic impact in the investigation of anti-icing agents in the laboratory. There was executed the comparative characteristics of the results of the preliminary pilot studies on the phytotoxic properties of IMR under using different substrates for germination of seeds - soil and filter paper. The data obtained are characterized by differences in the degree of phytotoxic action of the same species depending upon ice-melter reagents methodical setting circuit laboratory experiment. As a result, there was shown the imperfection of the existing method of rapid analysis in relation to ice-melter materials (IMM).

[The anemic syndrome in patients with prevalent stages of Hodgkin's lymphoma under application of intensive chemotherapy according the scheme EACOPP-14.]

The sampling of 61 patients with prevalent stages of Hodgkin's lymphoma at various stages of chemotherapy became subject of clinical blood analysis and evaluation of anemic syndrome. The anemia of I-IV degrees was diagnosed in all patients with Hodgkin's lymphoma. Among them patients with anemia of chronic disease prevailed. The single cases of iron-deficient and auto-immune hemolytic anemia were marked. In 24 patients with Hodgkin's lymphoma (39.3%) the deficiency of endogenous erythropoietin which was marked in both patients with anemia of chronic disease and patients with iron-deficient anemia. The lower level of endogenous erythropoietin was fixed in patients with leukopenia and very low erythropoietic activity of marrow. In 8 patients (13.1%) functional deficiency of iron against the background of anemia of chronic disease was established. Therefore, anemic syndrome in patients with prevalent stages of Hodgkin's lymphoma both inactive phase of disease and during application of chemotherapy represents complicated complex of symptoms. It requires accurate laboratory analysis and individual approach in treatment that in turn will favor optimal efficiency of therapy.

[If digital clubbing appears bear Marie-Bamberger disease in mind]. / Bei neu aufgetretenen Trommelschlegelfingern daran denken: Marie-Bamberger-Syndrom.

Secondary hypertrophic osteoarthropathy, also known as Marie-Bamberger disease, occurs in up to 5 % of patients with non-small cell lung cancer (NSCLC). If the syndrome constellation of finger clubbing and bone pain on palpation is recognized early, lung cancer can be diagnosed at an early stage. This article reports the case of a 52-year-old male patient with knee pain, distal edema, watchglass nails and finger clubbing which first appeared in August 2013. Following angiologic, cardiologic and orthopedic consultations radiographs of the hands, feet and lungs were taken. Following the presumptive diagnosis of hypertrophic osteoarthropathy, the patient was referred to a specialist pulmonary clinic where an adenocarcinoma of the lungs was diagnosed.

[The characteristics of anemic syndrome in patients with prevalent phases of Hodgkin lymphoma before treatment].

The study was carried out to analyze peripheral blood covering 23 patients with prevalent phases of Hodgkin lymphoma received no treatment previously. The clinical analysis of blood was implemented using hematologic analyzer Sysmex XE-2100. The content of ferritin, soluble receptors of transferrin and erythropoietin was estimated using enzyme-linked immunosorbent assay. The free hemoglobin was estimated using hemoglobin cyanide method. The direct Coombs test, counting of leukogram and analysis of morphology of erythrocytes were applied to all patients. The anemia is diagnosed in 19 patients (83%). In 18 out of them the anemic syndrome corresponded to anemia of chronic disease and in one patient asiderotic anemia was established. The rest of patients had no anemia. The anemia of chronic diseases characterized by microcytosis and hypochromia of erythrocytes, inadequate degree of anemia by production of erythropoietin and functional deficiency of iron in most of the patients. It should be emphasized that anemia of chronic diseases commonly is normocyte normochrome anemia whereas in patients with prevalent phases of Hodgkin Iymphoma the microcyte hypochrome anemia was detected before the treatment and it was followed by functional deficiency of iron. In patients without anemic syndrome microcytosis and hypochromia of erythrocytes were marked too. The techniques of laboratory diagnostic objectively reflecting iron metabolism (evaluation of level of soluble receptors of transferrin andferritin) and appropriate hormonal response to degree of anemia (production of erythropoietin) are to be included into algorithm of treatment of patients. The purpose is to timely detect type of anemia with the purpose of its appropriate correction prior to treatment onset.

[SCIENTIFIC EVIDENCE FOR HYGIENIC STANDARDS HARMONIZED WITH INTERNATIONAL RECOMMENDATIONS FOR PRIORITY POLLUITIONS OF SOILS].

In accordance with international approaches the ranking of permissible levels of chemicals in the soil for the Russian Federation was performed with the use of actual maximum permissible concentration (MPC), there were scientifically substantiated 33 standardsfor 26 substances and their combinations in the soil, differentiatedfor 6 groups offunctional areas for soil use. There is presented the algorithm determining the procedure for the establishment of MPC for unstudied chemicals for various functional areas of soil of populated areas, as well as draft of amendments to existing sanitary-hygienic regulations prior to processing the latter in accordance with modern requirements.

[Methodical approaches to the substantiation of hygienic requirements for the application of deicing materials].

In the paper there is presented a review of existing guidelines and rules of operation of deicing materials (DIM), as well as opportunities for their processing with the aim to ensure the security for the environment and public health. There are described the ecological- hygienic and toxicological properties of chemicals. There are reported results of a pot experiment for the assessment of the impact of DIM on the lawn.

[The algorithm of modern laboratory diagnostics of anemic syndrome in oncologic patients].

The algorithm of analysis of blood of oncologic patients with anemic syndrome was elaborated using large clinical sampling (n = 284). It is established that 72% of oncologic patients developed anemia by kind of chronic diseases anemia. However in some patients the anemic syndrome was caused by the iron deficiency The cases of B12 deficient, autoimmune hemolytic anemia were registered and inefficient erythropoiesis under metastatic lesion of bone marrow and myelodysplasia syndromes as well. The high importance of methods of objective evaluation of iron resources (level of ferritin and soluble receptors of transferrin), erythropoietic activity of bone marrow (reticulocytes and reticulocyte indices) and adequate response to anemia degree (level of endogenic erythropoietin). The possibility to detect the concentration of hemoglobin in reticulocytes makes it possible to monitor the restoration of hemoglobin content on early stages of treatment of patient with iron deficiency anemia.

[Changes of trace element content in MARC-145 cells before and after infection with virulent and attenuated strains of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV)].

The influence of virulent and attenuated strains of Porcine Reproductive and Respiratory Syndrome Virus with defined genomic differences on trace element profile of MARC-145 cells was investigated to elucidate the process of infection in vitro. The concentrations of seven trace elements (Al, Cr, Mn, Ni, Fe, Cu, Zn) in infected and non-infected cells were determined. Although viral replication was similar in all cases, there were distinct features of difference in the trace element profiles (changes of concentrations of Ni, Mn, Fe) in infected cells. In particular, cells infected with the attenuated strain of PRRSV (NADC8-251) contained very low level of Ni, compared with its pathogenic counterpart NADC8-252K. The infection with the Lelystad strain also led to a moderately high level of this trace element.

Neutron flow exposure as a test for survival of Artemia salina spores.

Live and heat-inactivated Artemia salina spores (samples with the same mass and filling density) were exposed to a flow of thermal neutrons from a (252)Cf radioactive source at an equivalent dose power of about 1 microSv/h. Irradiation led to a 4-fold acceleration of nauplius development and to modification of the element profiles of live spores. The difference between absorption/diffusion of thermal neutrons by live and dead spores was revealed.