Synthesis and Aminomethylation of 2-Amino-4-(2-chlorophenyl)-6-(dicyanomethyl)-1,4-dihydropyridine-3,5-dicarbonitrile N-Methylmorpholinium Salt.

Sequential reaction of 2-chlorobenzaldehyde, cyanothioacetamide, and malononitrile dimer in the presence of an excess of N-methylmorpholine resulted in the formation of N-methylmorphlinium salt of 2-amino-4-(2-chlorophenyl)-6-(dicyanomethyl)-1,4-dihydropyridine-3,5-dicarbonitrile. The resulting salt reacts under Mannich conditions with primary amines and an excess of formaldehyde to form substituted 2-alkylamino-4-(dicyanomethylene)-3,7-diazabicyclo[3.3.1]non-2-ene-1,5-dicarbonitriles. Structure of the key compound was confirmed by single crystal X-ray diffraction analysis.

N,N'-Diphenyldithiomalonodiamide: Structural Features, Acidic Properties, and In Silico Estimation of Biological Activity.

The spectral characteristics of dithiomalondianilide (N,N'-diphenyldithiomalonodiamide) were studied, and the dissociation constant was determined by potentiometric titration. Quantum-chemical methods at the B3LYP-D3BJ/6-311+G (2d,p) level were used to calculate the molecular geometry and vibrational spectra of the most stable tautomeric forms of dithiomalondianilide. The bioavailability parameters were calculated, and possible protein targets were predicted by the protein ligand docking method.

[Line immunoassay for detection of IgG antibodies to hepatitis E virus (Hepeviridae, Orthohepevirus, Orthohepevirus A)].

INTRODUCTION: The diagnostic efficacy of methods for hepatitis E serodiagnostic varies over a wide range; therefore, the combined use of tests of various formats is recommended. The aim of the research was to develop a test system for the detection of IgG antibodies to hepatitis E virus (HEV) in human serum by linear immunoassay (LIA). MATERIAL AND METHODS: Serum samples from patients with hepatitis and healthy individuals were tested using commercial enzyme-linked immunosorbent assay systems for the presence of IgG antibodies to viral agents causing hepatitis and other infections associated with liver pathology. Recombinant antigens ORF2 and ORF3 of HEV genotypes 1 and 3 were used. The "RecomLine HEV IgG/IgM" reagent kit (Mikrogen GmbH, Germany) was used as a comparison test system. RESULTS: The first Russian diagnostic kit "Blot-HEV", designed to detect IgG antibodies to individual HEV proteins in human serum using LIA, was developed. The antigenic base is represented by strips of a nitrocellulose membrane with immobilized recombinant antigens ORF2 (aa 406-660) and ORF3 (aa 1-113) of HEV genotypes 1 and 3, and control antigens in the form of discrete lines. The conjugate was mouse monoclonal antibodies to human class G immunoglobulins labeled with horseradish peroxidase. The chromogen solution contained the 3,3',5,5'-tetramethylbenzidine. A visual and digital recording of results was provided. The analytical sensitivity of the test kit was 0.625 IU/ml for ORF2 antigens and 2.5 IU/ml for ORF3 antigens. The absence of the influence of endogenous interfering substances on the results of the analysis and the absence of cross-reactions with antibodies to hepatitis pathogens of the other etiologies had been shown. The sensitivity of the test system compared to the "RecomLine HEV IgG/IgM" kit was 92%, specificity 97%. Shelf life in condition of storage was determined to be 12 months. CONCLUSIONS: The developed test can be used to confirm the results of ELISA in laboratory diagnosis of hepatitis E.

[The estimation of the hepatitis E proportion in the etiological structure of acute viral hepatitis in certain regions of of Kyrgyzstan.]

Despite the fact that the Kyrgyz Republic (KR) belongs to the highly endemic regions of the world for hepatitis E, the true extent of the spread of this infection in the country remains poorly understood. It was estimated the prevalence of serological markers of hepatitis E virus (HEV) infection among patients with acute viral hepatitis (AVH) from the regions of the Kyrgyz Republic with a high level of seroprevalence previously established by us. Blood sera samples of hepatitis patients who were admitted to hospitals of Kyrgyzstan in the period 2018-2019 were examined by the enzyme immunoassay method using the kits «DS-ELISA-Anti-HEVIgG¼ and «DS-ELISA-ANTI-HEV-IgM¼ (RPC Diagnostic Systems, Russia). IgG and IgM antibodies to HEV were detected in 103 of 344 studied samples (29.9%). Most often, seropositive specimens were detected among people of age groups under 20 and over 40 years old. Hepatitis with the fecal-oral mode of transmission was dominated in the structure of AVH: the specific gravity of hepatitis E was 47.9%, hepatitis A - 35.32%. Markers of mixed infections with other hepatitis viruses have been detected in 40.4% IgM-positive individuals. Thus, high prevalence of serological markers of HEV infection in the territory of Kyrgyzstan during the interepidemic period had been shown. The necessity of including the determination of serological markers of hepatitis E into the algorithm for the comprehensive diagnosis of AVH in patients of all age groups with liver pathology had been confirmed.

[The analysis of sero-prevalence of virus of hepatitis E among patients with HIV-infection and syphilis.]

The infection with hepatitis E virus is one of causes of liver diseases in patients with secondary immunodeficiency, including HIVinfected ones. The study was carried out concerning analysis of rate of detection of serological markers of infection with hepatitis E virus in samples of blood serums of HIV-infected patients and other group of patients with expressed immuno-suppression - patients with syphilis. The sero-prevalence of hepatitis E virus on IgM-antibodies among HIV-infected patients in Moscow made up to 21.7% and 2.9% correspondingly. In the examined group from the Far-East region the highest sero-prevalence of hepatitis E virus on specific IgE-antibodies (73%) is established in the group of patients infected with HIV subtype B. The most frequently IgG and IgM antibodies to hepatitis E virus were detected in HIV-infected patients at the stage of disease 4B. The examined group of patients with syphilis the sero-prevalence of hepatitis E virus made up to 14.7% that significantly surpasses the given indicator in the group of healthy persons (1.7%). The increasing of the level of sero-postivity is demonstrated in the groups of patients with latent syphilis and on the second stage of disease. Therefore, the highest values of sero-prevalence of hepatitis E virus is observed in the groups of HIV-infected patients and patients with syphilis at late stages of disease. In the Russian Federation, the identification of antibodies to hepatitis E virus in HIV-infected patients depends of the region of residence.

[Pathomorphological peculiarities of hemomicrocirculatory bed of the small and large intestine in acute peritonitis].

Experimental comparative morphological investigation of hemomicrocirculation bed (HMCB) of the small and large bowel wall was performed in dynamics of an acute serous peritonitis. Spreaded aseptic peritonitis was simulated using injection of 5 ml of gamma-caraginen (Sigma, USA) in 1 ml of isotonic solution of sodium chloride. On the early stage of peritonitis (in 12 h from beginning of the experiment) in mucosa of small bowel nonsignificant venuls dilatation and the capillary lumen reduction were observed. In 1 day (reactive stage of peritonitis) in mucosa the quantity of capillars have had reduced significantly, comparing with such observed previously. On the 2-nd day (toxic stage of peritonitis) some capillary dilatation in intestinal villi and crypts coexistant with the blood rheology disorders in a form of stasis, change in permeability of the vessels walls, predominantly of the venous, was noted. On the 3-d day (late stage) the arteriol's spasm have had reduced, capillary paralytic dilatation was revealed. The staged course of experimental peritonitis with the HMCB changes, characteristic for every stage, was confirmed, basing on analysis of the investigation result.

[Immune status of patients after splenectomy, performed for the spleen traumatic injury in remote terms and choice of preparations for its correction].

Immune status was analyzed in 28 patients in remote period after splenectomy. In accordance to immunological data obtained in splenectomy patients, inhibition of cellular chain of speciphic and natural immunity was noted on background of hyperactivation of its humoral chain with signs of autoimmune processes. In such patients, while immunomodulators selection, the mostly pronounced stimulating impact was noted in immunoglobulin; of sorbents the most binding capacity has rheopolyglukin. This opens further perspectives of postsplenectomy patients treatment, using immunomodulating and elimination correction of the disorders revealed. Expediency of application of rheopolyglukin and immunoglobulin in contents of eliminating amed immunomodulating therapy was substantiated. Studying of peculiarities of immune status constitute a perspective way in the patients, in whom various variants of organpreserving tactics while surgical interventions on spleen were applied.

[Estimation of the results and quality of life indices in patients with gastroduodenal ulcer depending on the method of operative treatment].

The results of operative treatment of 261 patients, suffering perforative gastroduodenal ulcer, are analyzed. In 159 patients the ulcer excision with pyloroplasty and without vagotomy was performed and in 102--the ulcer excision with vagotomy. In all the patients the chronic ulcer perforation was noted. In operated patients gastric mucosa have had morphologic peculiarities, characteristic for chronic atrophic gastritis with complete intestinal metaplasia foci. In surgical treatment for perforative gastroduodenal ulcer its excision with pyloroplasty conduction and medicinal correcting therapy are recommended.

[Prevalence of antibodies to hepatitis E virus among residents of different climate and geographic zones of Russia Federation]. / Chastota vstrechaemosti antitel k virusu gepatita E u naseleniia razlichnykh klimatogeograficheskikh zon Rossiiskoi Federatsii.

The detection frequency of antibodies to Hepatitis E Virus (HEV) was studied in residents of the South and of the Middle European Part of the Russia Federation as well as of Siberia. Antibodies to HEV were most often found both in patients with hepatic pathologies and in subjects with diseases unrelated with a primary hepatic lesion, in particular, in patients with skin and venereal diseases and with HEV. A higher concentration of antibodies to HEV was noted also in blood donors, medical personnel and isolated communities, like prisons or psychiatric clinics. A correlation was established between the rate antibodies to HEV are registered and such risk factor as contacting with blood or a gross violation of the hygienic rules.

[Propagation of hepatitis E virus in an endemic and nonendemic regions]. / Osobennosti rasprostraneniia virusa gepatita E v éndemichnom i neéndemichnom regionakh.

The role of hepatitis E (HE) in sporadic morbidity at an endemic territory (Southern Uzbekistan) and the incidence of anti-HEV in different populations of a nonendemic region (Russia) were evaluated. Antibodies to HEV were detected in 22.1% of patients with acute HE, including mixed infections (+ HA or HB), in the Dekhkanabad district of Kashkadarya region in 1993. The estimated incidence of acute HE was 51.7 per 100,000 population. Analysis of monthly incidence of acute HE demonstrated a seasonal pattern of the morbidity: more than 80% of total recorded cases occurred in August-September. These data indicate the presence of group HEV infections and an important role of this infection in the structure of acute viral hepatitis at the endemic territory. Anti-HEV were found in some population groups at a nonendemic territory: in free-of-charge blood donors in Surgut (4%), in patients with HIV infection (1.6%), and in medical workers in Moscow (1.1%).