[Myorenal syndrome in forensic practice: molecular aspects of etiology and pathogenesis]. / Miorenal'nyi sindrom v sudebno-meditsinskoi praktike: molekulyarnye aspekty etiopatogeneza.

The objective of the study is to analyze the publications on biochemical aspects of myorenal syndrome (crush-syndrome) pathogenesis. Factors of trauma and other etiologies significant in terms of forensic practice that cause muscle tissue destruction are presented. Molecular processes in rhabdomyolysis and subsequent renal damage, the establishment of the sequence of which is important for forensic medicine, are outlined. The study results will improve our understanding of the of myorenal syndrome pathophysiology, its biochemical features, and optimize methods for its forensic diagnosis.

Genetic resources of narrow-leaved lupine (Lupinus angustifolius L.) and their role in its domestication and breeding.

Narrow-leaved lupine (Lupinus angustifolius L.) is a cultivated multipurpose species with a very short history of domestication. It is used as a green manure, and for feed and food. This crop shows good prospects for use in pharmacology and as a source of f ish feeds in aquaculture. However, its genetic potential for the development of productive and adaptable cultivars is far from being realized. For crop species, the genetic base of the cultivated gene pool has repeatedly been shown as being much narrower than that of the wild gene pool. Therefore, eff icient utilization of a species' genetic resources is important for the crop's further improvement. Analyzing the information on the germplasm collections preserved in national gene banks can help perceive the worldwide diversity of L. angustifolius genetic resources and understand how they are studied and used. In this context, the data on the narrow-leaved lupine collection held by VIR are presented: its size and composition, the breeding status of accessions, methods of studying and disclosing intraspecif ic differentiation, the classif ications used, and the comparison of this information with available data on other collections. It appeared that VIR's collection of narrow-leaved lupine, ranking as the world's second largest, differed signif icantly from others by the prevalence of advanced cultivars and breeding material in it, while wild accessions prevailed in most collections. The importance of the wild gene pool for the narrow-leaved lupine breeding in Australia, the world leader in lupine production, is highlighted. The need to get an insight into the species' ecogeographic diversity in order to develop cultivars adaptable to certain cultivation conditions is shown. The data on the testing of VIR's collection for main crop characters valuable for breeders are presented. Special attention is paid to the study of accessions with limited branching as a promising gene pool for cultivation in relatively northern regions of Russia. They demonstrate lower but more stable productivity, and suitability for cultivation in planting patterns, which has a number of agronomic advantages. Analyzing the work with narrow-leaved lupine genetic resources in different national gene banks over the world helps shape the prospects of further activities with VIR's collection as the only source of promising material for domestic breeding.

ARTERIAL CHEMOINFUSION IN PATIENTS WITH LOCALLY ADVANCED AND METASTATIC PANCREAS CANCER.

The chemoinfusions (310) were carried out in celiac trunk in 167 patients with non-removed pancreas cancer at the period from 2000 to 2015. Locally advanced timorous process (stage III, n=79) was revealed in 79 patients and liver metastases (stage IV, n=88) were noted in 88 cases. The celiac axis infusion by Gemcitabine (1000 mg/m²) was applied for patients and GEMOX (Gemcitabine+Oxaliplatin 75 mg/m²) has been using since 2012. Symptomatic improvement such as decrease of pain, growth of body weight was noted in majority of patients. An average lifetime, median and one-year survival consisted of 7,6 months, 5,8 months and 10%. The patients (133) were treated by 1­2 cycles and after that by course of total body chemotherapeutics. There weren't any serious complications. Toxic manifestations of chemotherapy weren't higher than I­II degree and they were arrested by corrective therapy in 92 patients (55%). The celiac axis infusion is safe in patients with locally advanced and inoperable pancreas cancer. Symptomatic improvement showed the most patients. The objective response to the treatment had 20% patients and performance of repeated cycles led to increase of their survival.

[Palivizumab: four seasons in Russia].

In 2010, the Russian Federation (RF) registered palivizumab--innovative drug, based on monoclonal antibodies for passive immunization of seasonal respiratory syncytial virus (RSV) infection in children of disease severe progress risk group, which include primarily premature infants, children with bronchopulmonary dysplasia and hemodynamically significant congenital heart disease. Currently, palivizumab is included in the list of recommended medicines and medical care standards of different countries, including Russia. In the review the results of Russian research on the progress of RSV infection, its epidemiology and immunization experience gained over the 2010-2014 period are summarized in relation to the foreign data. During the four epidemic seasons palivizumab immunization covered more than 3,200 children of severe RSV infection risk group with a progressive annual increase in the number of patients who received the drug. Geography of palivizumab immunization is also greatly expanded in our country during this time. If during the first two seasons measures of immunization were taken mainly in Moscow and St. Petersburg, at the present time, thirty one territorial entities of the Russian Federation have the experience in the drug application. Analysis of the results of RSV infection immunization (made in several regions) confirms the high clinical efficacy and palivizumab safety already demonstrated in international studies. In addition, the analysis presents the potential to improve the efficiency of the integrated RSV infection immunization programs, realizing in the establishment of high-risk child group register, adequate counseling for parents, as well as the development of the routing of patients and coordination of interaction between different health institutions during the immunization.

[Biological properties of salmonellae and local immunity in Salmonella infection].

The comparative study of the biological properties of S. enteritidis and S. typhimuruim, isolated from patients and convalescent carriers, was carried out. Factors inactivating the components of the local immunity of the intestine (lysozyme, complement, lactoferrin, IgG, IgM and IgA) were detected in the causative agents of Salmonella infections. The spread and expression degree of properties of a causative agent were serovar-depended: high penetration characteristics and the expression of anti-lactoferrin and anti-immunoglobulin activity were characteristic of S. typhimurium. S. enteritidis strains isolated from patients with carrier state formed in the convalescence period were found to have higher persistence level. In co-profiltrates obtained from carriers at the peak of the disease and during convalescence lower levels of IgM, IgG, sIgA, complement and lactoferrin were determined in comparison with those in coprofiltrates obtained from patients in whom no subsequent carrier state was formed. These results indicate that an increase in the persistence of salmonellae, occurring simultaneously with the local immunodeficiency, contributes to the prolonged survival of bacteria in the intestine.

Association of prognosis in surgically treated lung cancer patients with cytometric, histometric and ligand histochemical properties: with an emphasis on structural entropy.

OBJECTIVE: To explore new tumor features for refined category formation that permits the tailoring of individualized treatment schemes in lung cancer. STUDY DESIGN: Survival data on patients from six independent studies on cases with surgically treated lung cancer, primary lung carcinoids or metastasizing breast carcinoma (including data on primary breast carcinoma) were analyzed by nonhierarchic multivariant discriminant analysis with respect to a set of cytometric/histometric and immunohistochemical/ligand histochemical parameters. The number of stem lines, S-phase-related tumor cell fraction and the extent of structural entropy and its current were measured. In addition, the expression of binding capacities for histo-blood group trisacharides, galectins, the alpha/beta-interferon antagonist sarcolectin, the lymphokine macrophage migration inhibitory factor and a monoclonal antibody to the Le(y) epitope was monitored for insight into aspects of immunologic and biologic behavior. RESULTS: In all studies, a correlation between tumor parameters, according to TNM stage and survival, was seen. In order to refine this category formation, at least certain selected features should provide an even more stringent association than TNM stages. Indeed, statistical correlation of the cytometric and histometric parameters as well as the expression of receptors for the two histo-blood group trisaccharides, ligands for the galectins (CL-16, CL-14) and macrophage migration inhibitory factor was stronger than that of TNM stage. A large amount of the current of structural entropy was especially highly significantly associated with poor survival. This observation could be verified in each of the different studies. CONCLUSION: The obtained data strongly support the notion that thermodynamic evaluation of tumor growth focusing on the "entropy distance" of the tumor from its environment is a promising perspective warranting extended studies. Additionally, glycohistochemical features, including binding capacities for histo-blood group trisaccharides, have the potential to aid in establishment of a biologic marker set for tumor staging.

[Study of fluorescence from isolated yeast plasma membranes in visible range of the spectrum]. / Issledovanie fluorestsentsii izolirovannykh plazmaticheskikh membran drozhzhei v vidimoi oblasti spektra.

The fluorescence with maximum at 683 nm from isolated yeast plasma membranes has been detected. The fluorescence was due to a membrane-bound compound absorbing in the visible range of the spectrum. The fluorescence excitation spectrum of this compound has a structure typical for porphyrins. At the same time, in several fluorescence properties the porphyrin localized in the plasma membrane is different from other intracellular porphyrins (protoporphyrin, coproporphyrin).

Summary report on the ISOBM TD-6 workshop: analysis of 20 monoclonal antibodies against Sialyl Lewisa and related antigens. Montreux, Switzerland, September 19-24, 1997.

The ISOBM TD-6 Workshop is the first international workshop on monoclonal antibodies against the Sialyl Lewisa (SLea) antigen. Eight research groups participated in a blind study to characterize the epitope binding, relative affinity and performance in immunoradiometric assays, of a panel of 20 monoclonal antibodies. The antibodies were tested against a diverse panel of neoglycoconjugates, purified antigens and human serum pools from gastrointestinal malignancies. Epitope specificities were determined for the majority of antibodies in the panel. Cross-reactivity with related saccharide structures was noted in several antibodies. Overall, the results of the TD-6 Workshop show further development of SLea immunoassays may yield yet more specific assays for the detection and management of gastrointestinal and other malignancies.

[Selectin receptors. 5. Monoclonal antibodies to the synthetic antigens SiaLe(a) and SiaLe(x)]. / Retseptory selektinov. 5. Monoklonal'nye antitela k sinteticheskim antigenam SiaLea i SiaLex.

Five IgM class monoclonal antibodies (MAb) to the SiaLea tetrasaccharide, which is known as a serologic tumor marker CA 19.9, and three MAbs (one of IgG3 and two of IgM class) to the SiaLex tetrasaccharide (differentiation antigen CD15s) were obtained against totally synthetic immunogens. The epitope specificity of the antibodies was determined using a wide range of oligosaccharides and their polyacrylamide conjugates. MAb 4E10 against SiaLea and MAb 4G5 to SiaLex were highly specific to the antigen predefined by immunization: they did not cross-react either with structurally and conformationally related oligosaccharides or with their disaccharide fragments. Two MAbs to SiaLea (D7 and E5B1) showed a weak binding to SiaLex. MAb CC1 recognized SiaLea and SiaLex almost equally, and MAb 5H9 to SiaLea cross-reacted with the non-sialylated form, the Lea trisaccharide. Two MAbs against SiaLex A3 and B11 bound to all carbohydrate structures containing the core disaccharide Gal beta 1-3(4)GlcNAc.

Monoclonal antibody LU-BCRU-G7 against a breast tumour-associated glycoprotein recognizes the disaccharide Gal beta 1-3GlcNAc.

The monoclonal antibody LU-BCRU-G7, that was generated by in vitro immunization, shows clinical value as a prognostic marker in early stage breast carcinoma. It has now been characterized with regard to its binding epitope. Using a recently described method based on the construction of N-substituted polyacrylamide (PAA) derivatives of carbohydrates (pseudopolysaccharides), the structure of the epitope for the monoclonal antibody LU-BCRU-G7 has been determined. Competitive binding assays and inhibitory enzyme-linked immunosorbent assays (ELISAs) using these pseudopolysaccharides have shown the LU-BCRU-G7 epitope to be a disaccharide Gal beta 1-3GlcNAc (Lec; where Gal is D-galactose, Glc is D-glucose and GlcNAc is N-acetyl-D-glucosamine). Both galactose and N-acetyl glucosamine moieties are essential for binding. Substitution on C-2 or C-3 of the terminal galactose abolished binding, as did galactose-alpha terminated oligosaccharides. The galactose moiety alone, as expressed by the Gal beta-PAA conjugate, appeared to be a more important feature of the epitope than the GlcNAc-PAA conjugate, which failed to bind or inhibit the LU-BCRU-G7 antibody. In the N-acetyl glucosamine moiety, binding was decreased but not eliminated by fucose substitution, as in Lea, or change in configuration of C-4, as in Gal beta 1-3GlcNAc. Omission of the NAc group resulted in complete loss of activity. The tetrasaccharide lacto-N-tetraose, although containing the terminal Lec disaccharide, does not react with the antibody, suggesting conformational interference of the binding site.(ABSTRACT TRUNCATED AT 250 WORDS)

Purification of monoclonal antibodies to Le(y) and Le(d) carbohydrate antigens by ion-exchange and thiophilic-adsorption chromatography.

The paper deals with convenient and fast methods for purification of monoclonal antibodies (MAbs) to carbohydrate antigens Le(y) and Le(d) from the cell culture and ascite fluid by ion-exchange chromatography on S-Sepharose and salt-promoted chromatography on a "thiophilic" adsorbent. One-step procedure on S-Sepharose of MAbs to Le(y) (IgG and IgM) provides significant purification (over 90% of contaminants were removed), while a purification factor for IgM to Le(d) is much lower. Highly purified IgM to Le(d) could be obtained by a two-step purification procedure including "thiophilic-adsorption" chromatography and gel-filtration (90-98% of contaminants from the cell culture and ascite fluid were removed). The preparations of IgG and IgM retain their initial antibody activity.

Monoclonal antibodies directed to the synthetic carbohydrate antigen Ley.

Tetrasaccharide Fuc alpha 1-2Gal beta 1-4(Fuc alpha 1-3)GlcNAc is known as carbohydrate determinant of cancer- and AIDS-associated antigen Lewisy (Ley). Synthetic antigen to generate mouse monoclonal antibodies (MAbs) directed to Ley was prepared and constructed as a spacer-armed tetrasaccharide coupled with lipophilized polymer, Ley-PAA-PE, where PAA is a 30-kD polyacrylamide and PE is phosphatidylethanolamine. An efficient immune response was provided by using Ley-PAA-PE adsorbed on Salmonella minnesota. Positive hybridomas were screened by enzyme-linked immunosorbent assay (ELISA) using Ley-PAA as a coating agent. An inhibitory version of the same test system showed absolute specificity of two MAbs: only hapten Ley and Ley-PAA were strong inhibitors, in contrast to Leb, tri- and disaccharidic fragments of the mentioned tetrasaccharides, as well as their PAA-conjugates. MAbs obtained against synthetic antigen specifically stained the Ley (+) cell line A431.

On the in vitro neutralization test of Clostridium perfringens type A toxin.

Data are presented on the detection in crude animal and human sera of Cl. perfringens phospholipase C (PLC) inhibitor. When the level of Cl. perfringens type A antitoxin is determined in the in vitro toxin neutralization test the inhibitor is found to decrease PLC activity in the test dose of experimental homologous toxin. The extent of decrease accounts for the variation of results obtained in the in vitro and in vivo toxin neutralization tests. The variation may be cancelled out by introducing a corresponding coefficient to calculate the level of alpha-antitoxin. It is suggested that the isolation and investigation of the PLC inhibitor will contribute to the development of preparations for treatment of gas gangrene due to Cl. perfringens type A.

Comparative study on the immunogenic properties of Clostridium perfringens type A toxoid.

The paper presents the results of a study on the immunogenic properties of toxoid preparations from Cl. perfringens type A obtained using the routine method of detoxifying alpha = toxin in the culture medium (commercial preparations) and by means of detoxifying a previously purified alpha = toxin (experimental preparations). When tested in immunized guinea pigs, the immunogenicity of experimental preparations was found to be 4.5 to 6 times that of commercial preparations. In mice, there was no difference in the immunogenic properties of the two types of preparations as determined by the ED30 of the antigen and the serum levels of Cl. perfringens antitoxin. The possibility is discussed of using the guinea pig as a laboratory animal model due to its ability to reflect most clearly the differences in the immunogenicity of Cl. perfringens type A toxoid preparations.

[Comparative evaluation of the methods of determining perfringens A antitoxin]. / Sravnitel'naia otsenka metodov opredeleniia urovnia antitoksina perfringens tipa A.

The methods for determining the level of type A Perfringens antitoxin in human blood sera were examined and compared. The ratios for correlating the data obtained in the toxin neutralization test (NT) in vivo, in the passive hemagglutination test (PHT), and as a result of the enzyme-labeled immunosorbent assay (ELISA) with regard to the antitoxin level measured in the NT in vitro were equal to 0.88, 0.64 and 0.39, respectively. The sensitivity of the NT in vivo and in vitro was 0.25 IU/ml, that of the PHT 0.01-0.005 IU/ml, and that of the ELISA 0.01-0.02 IU/ml Perfringens antitoxin. To perform the NT, not less than 1 ml blood serum is required, while for the PHT and ELISA, 0.1-0.05 ml. Provided hyghly purified anatoxin is used for preparing the erythrocyte diagnosticum Perfringens, and polysterene plates are sensitized in performing the ELISA, all the reactions are specific. While titrating human blood sera containing type A Perfringens antitoxin, use in the PHT may be made of type A Perfringens rabbit antiserum as reference.

[Factors modulating production of antibodies against Clostridium perfringens in mammals]. / Faktory, moduliruiushchie antitelogenez v otnoshenii anatoksina Clostridium perfringens u mlekopitaiushchikh.

Immunogenic properties of Clostridium perfringens type A toxoid preparations obtained by different methods are described. As regards immunogenicity for guinea-pigs and man, toxoid obtained by the detoxification of preliminarily purified alpha-toxin (experimental toxoid) compares favourably with preparations obtained by the detoxification of alpha-toxin in a culture fluid. It was shown in experiments on guinea pigs that immunogenicity of experimental toxoid rises with the increase in the degree of purification of alpha-toxin used for detoxification. The least purified preparations turned out to be the most immunogenic for mice. In Cl. perfringens culture fluid, factor (factors) was found and separated, which reduced the immunogenicity of the respective toxoid in experiments with immunization of guinea pigs and increased it in mice. Activation of antibody-formation was disclosed to be nonspecific. The existence of this factor(s) should be taken into account in the development of vaccines and methods for evaluating their immunogenic properties.

[Characteristics of the immunologic response of animals to Clostridium perfringens anatoxin]. / Osobennosti immunnogo otveta zhivotnykh na anatoksin Cl. perfringens.

Experiments were conducted on guinea pigs, rabbits and mice (mongrel and inbred); immunogenic properties of Cl. perfringens toxoids of different purity were studied. Toxin neutralization and passive hemagglutination tests were used to determine the antitoxic immunity level. It appeared that in the immunization of guinea pigs and rabbits the degree of immunogenicity of the preparations increased with the elevation of their specific activity. Under the same conditions both the mongrel and the inbred mice displayed the maximum immune response in the immunization with the least purified preparations, and the minimum after the injection of a highly purified antigen.

[Properties of C1. perfringens toxoids obtained from purified toxins]. / Svoistva anatoksinov C1. perfringens, poluchennykh iz ochishchennykh toksinov

The authors present the results of comparative study of the properties of experimental perfringens toxoids obtained from purified alpha-toxoids of different degrees of purity. Experimental toxoids proved to possess a greater immunogenicity than preparations obtained by detoxication of alpha-toxin under conditions of cultural fluid, the greater--the more the purity of alpha-toxin used for procuring the experimental toxoid. C1. perfringens alpha-toxin recovered as a result of two-stage alpha-toxin purification, including its primary concentration and fractionation on DEAE-cellulose under conditions of negative alpha-toxin sorption, changed during detoxication into toxoid whose immunogenicity exceeded that of manufactured preparations 3--4 fold. The toxoid was harmless and sorbed in a dose of 100 BU on 2--3 mg of aluminium hydroxide; stability of its antigenic properties and its yield was not less than those of manufactured toxoids. Perfringens toxoid prepared from highly purified alpha-toxoid was 10 times greater by immunogenicity than the manufactured preparation, and was sorbed on 1--2 mg of aluminium hydroxide.

[Immune response of mice of different inbred lines to Clostridium oedematiens anatoxin]. / Immunnyi otvet myshei raznykh inbrednykh linii na anatoksin Clostridium oedematiens

Mice belonging to a number of inbred strains were immunized intradermally with Cl. oedematiens alpha-toxoid. The immunization was repeated 30 days later. On the 20th and the 30th days after the first injection and on the 10th day after the second one the antibody level against the toxoid was determined in the blood of mice by the passive hemagglutination test. The maximum response to the primary immunization was observed in the mice of the C3H strain, and the minimum one--in mice of the DBA/2 strain; the difference was more than 30-fold. The rest of the strains used in the test (A,CBA, BALB/c, AKR, CC57BR) displayed an intermediate level of the immune response. The differences reduced after the repeated immunization. The immune response to this antigen in mice is supposed to be genetically controlled.