Synthesis and Anti-HIV-1 Activity of Olean-9(11),12(13)-Dien-30-Oic Acid 3ß-(2-O-ß-D-Glucuronopyranosyl-ß-D-Glucuronopyranoside).

The glycyrrhizic acid (GA) analog olean-9(11),12(13)-dien-30-oic acid 3ß-(2-O-ß-D-glucuronopyranosyl-ß-D-glucuronopyranoside) (II) was synthesized via reduction of GA by NaBH4 in refluxing 2-PrOH:H2O with subsequent work up with HCl (5%). The cytotoxicity and antiviral activity of this glycoside against HIV-1 was studied in MT-4 cell culture. It was found that II was practically non-toxic for MT-4 cells while inhibiting accumulation of virus-specific protein p24 and RNA-dependent DNA-polymerase activity of HIV-1 reverse transcriptase (RT) (IC50 3.1 ±1.0 µg/mL).

[Correction of secondary hyperlipidemia in children with diabetes].

At 25 children with insulin dependent diabetes mellitus (IDDM, Type I) in a decompensation stage was estimated efficiency of correction secondary hyperlipidemia. The basic group (n = 11) along with insulinotherapy and a lipotrophic preparation (methionine) accepted probiotic Bifiform Baby in a dose 0.5 g once a day within 10 days; the comparison group (n = 14) received insulin therapy and a lipotrophic preparation. Efficiency of treatment was estimated on dynamics of lipids in serum of blood: triglycerides (TG), the total cholesterol (TH), betalipoproteides (BLP). At patients of the basic group authentic decrease TG, TH, BLP was established; at patients of group of comparison--decrease TH and BLP. Advantage of therapy secondary hyperlipidemia at children with diabetic ketoacidosis with use of Bifiform Baby consists in the elimination of the main infringement of a metabolismus of lipids in the given disease--hypertriglyceridemia.

[Development and certification of libraries of the MDCK continuous cell line for production of influenza vaccine].

Seeding and working banks of the continuous MDCK cell culture suitable for the production of cultured influenza vaccine were created and deposited at liquid nitrogen temperature at the "Vector" State Research Center of Virology and Biotechnology. The MDCK cell culture was shown to have morphology typical of the discussed cell line; it does not have any alien agents and is oncogenically safe; its enzimogram and karyotype are typical of the donor line; finally, its biological properties are stable during a long period of cultivation and its sensitivity to influenza virus is high, therefore, it can be recommended for the production of influenza vaccine. The continuous MDCK cell line was certified at Tarasevich Committee and was recommended by the MIBP Committee, Russia's Health Ministry, for its use as a substrate in the production of diagnostic and preventive immunoglobulins.

[The anti-HIV activity of glycyrrhizic acid penta-O-nicotinate]. / Anti-VICh-aktivnost' penta-O-nikotinata glitsirrizinovoi kisloty.

The anti-HIV activity of niglizin (penta-O-nicotinate of glycyrrhizic acid) and of its combinations was studied in the culture of infected MT-4 cells and in respect to the recombinant reverse HIV-1 transcriptase. Niglizin was shown to suppress effectively the HIV replication and to be a noncompetitive inhibitor of reverse transcriptase. Research of a combined anti-HIV action of niglizin and of azidothimidine (AZT) demonstrated that the preparations, when used at ratios of 1:20, 1:50, 1:200 and 1:2000, suppressed the synergetic effect both in the cell culture and in the recombinant reverse HIV transcriptase. A study of the antiviral activity of niglizin and of its joint use with AZT in respect to the AZT-resistant HIV-1 mutant showed niglizin to be more effective (ID50 = 0.134 microM) versus the "wild" strain. (ID50 = 9.64 micriM); whereas, its combined use AZT:niglizin = 1:100 displayed synergism (FIC = 0.553). The efficiency of the combined AZT/niglizin anti-HIV effect both in respect to the "wild" strain and to the AZT-resistant mutant confirms that such combinations are promising for the treatment of HIV infection.

[Determination of glycyrrhizic acid binding sites by a phage display method]. / Opredelenie uchastkov sviazyvaniia glitsirrizinovoi kisloty metodom fagovogo displeia.

Phages that expose peptides specifically interacting with glycyrrhizic acid (GA) were selected from a phage peptide library by affinity selection and ELISA. Amino acid sequence analysis of the selected peptides and human proteins with the SIM program revealed homology to tyrosine protein kinases, serine/threonine protein kinases, tyrosine phosphatases, and some receptors. Analysis of the peptide and virus protein sequences with the BLAST program showed that GA has affinity for various surface proteins of several human viruses such as HIV-1, hepatitis C virus, and herpesviruses.

Characterization of recombinant integrase of human immunodeficiency virus type 1 (isolate Bru).

Integration of the human immunodeficiency virus type 1 (HIV-1) DNA into the human genome requires the virus-encoded integrase protein. The recombinant integrase protein of HIV-1 (isolate Bru) was prepared by constructing a plasmid based on pET-15b encoding the integrase gene. Integrase of HIV-1 was purified using a bacterial expression system (Escherichia coli). The main kinetic parameters of HIV-1 integrase (K(m) = (3.7 +/- 0.2).10(-10) M, k(cat) = (1.2 +/- 0.3).10(-7 )sec(-1)) were determined using an oligonucleotide duplex constructed on the basis of the U5-terminal sequence of proviral HIV-1 DNA as the substrate. Inhibition of integrase by aurintricarbonic acid ([I](50) = 6.3 +/- 0.4 microM) and dependence of integrase activity on Mg2+ and Mn2+ concentration were studied.

[Nonpolar interactions in phospholipase A2 molecules]. / Nepoliarnye vzaimodeistviia v molekule fosfolipazy A2.

Nonpolar regions (hydrophobic nuclei and microclusters) in porcine pancreatic phospholipase A2 have been investigated and described. Based on the atomic coordinates, we analyzed all nonpolar contacts for all nonpolar groups of each amino acid residue and identified three hydrophobic nuclei (gamma 1, gamma 2, gamma 3) and four microclusters (mu 4, mu 5, mu 6, mu 7) in the enzyme. These nonpolar regions include hydrophobic residues as well as residues with charged or polar side chain. Quantitative characteristics of hydrophobic nuclei show that the latter are condensations of nonpolar contacts in the protein. The stability of the nuclei is comparable to that of the secondary structure elements. Comparative analysis of nonpolar areas in native porcine pancreatic phospholipase A2, mutant enzyme, and its complex with a substrate analog demonstrated the role of nonpolar contacts in protein structure and function.

[Bacterioscopy of cerebrospinal fluid from patients with generalized forms of meningococcal infection]. / Bakterioskopiia obraztsov spinnomozgovoi zhidkosti bol'nykh pri generalizovannykh formakh meningokokkovoi infektsii.

The author analyzes the potentialities of bacterioscopic examinations of cerebrospinal fluid samples of patients with meningococcal infection and correlations of the liquor appearance characteristics and bacteriologic data. The findings are extremely important for practical laboratories, for they permit an adequate assessment of bacteriologic examinations and improve their quality. Six CSF groups may be distinguished, differing in their type. A relationship has been revealed between liquor appearance and bacteriologic and bacterioscopic findings, this relationship being prognostically and diagnostically significant.

[Arterial hypertension in patients with alcoholic abstinence syndrome and delirium]. / Arterial'naia gipertenziia u bol'nykh s alkogol'nym abstinentnym sindromom i deliriem.

Arterial pressure, fear and anxiety affects, urinary excretion of catecholamines and plasma renin activity, central hemodynamics were studied in 134 subjects with alcoholic abstinence syndrome and delirium. The above syndrome and delirium were associated with arterial hypertension termed "reactive". Reactive hypertension declined 1-2 days after the patients' getting out of the acute stage of the affects. It can be considered as a manifestation of stress-response in pathogenesis of which an essential role may be played by plasma catecholamines and renin elevation. Hemodynamically, such hypertension is hyperkinetic with a tendency to high total peripheral resistance. Correlations were not found between the degree of the reactive hypertension and severity of the fear and anxiety affects. There was a weak correlation between the affects and heart beat. Apart from reactive hypertension, alcoholism is a promotor of essential hypertension.

[Assessment of the sensitivity of Neisseria meningitidis strains to benzylpenicillin using the disc diffusion method]. / Otsenka chuvstvitel'nosti shtammov Neisseria meningitidis k benzilpenitsillinu disko-diffuzionnym metodom.

The sensitivity of 235 N. meningitidis strains to 5 antibiotics was estimated by the diameter of growth inhibition zones according to the criteria recommended by a Laboratory (Marseilles, France) collaborating with the WHO. All the strains proved to be sensitive to benzylpenicillin when disks containing 10 and 2 IU of the antibiotic were used. The strains were also shown to be sensitive to chloramphenicol and tetracycline. 95.7 and 7.7 per cent of the strains were sensitive to rifampicin and oleandomycin, respectively. When the strain sensitivity was assayed with the disks containing 10 and 2 IU of benzylpenicillin by the more severe criteria recommended by J. Saez-Nieto et al., significant changes were detected: meningococci with relative resistance to benzylpenicillin were detected in various regions of this country and the number of such strains was found to have a tendency to slightly increase.

[The characteristics of suppurative meningitis morbidity caused by Streptococcus pneumoniae and Haemophilus influenza type B]. / Osobennosti zabolevaemosti gnoinymi meningitami, vyzvannymi Streptococcus pneumoniae i Haemophilus influenzae tipa B.

Of the 1,018 patients with purulent bacterial meningitis, hospitalized at the 2nd Clinical Infectious Hospital in Moscow during the period of 1980-1987, the diagnosis was confirmed in 54.7%; of these, meningitis of pneumococcal etiology was established in 44.8% and meningitis caused by H. influenzae, type b, in 23.8% of the patients. In meningitis of pneumococcal etiology high risk groups included mainly adults, especially those over 50 years, and children under 3 years of age. In meningitis of H. influenzae etiology high risk groups included mainly young children under 2 years of age. Meningitis of pneumococcal etiology was characterized by considerable death rate (on the average, 20%), while in meningitis of H. influenzae etiology death rate was 3 times lower. Pneumococci of serotypes 1, 3, 6, and 19 were found to be of the highest etiological importance for adults and pneumococci of serotypes 19, 6, 12, and 1, for children. In recent years greater etiological role of serotype 42 in adults was noted. The study of the spread of meningitides of different etiology is a high-priority task for this country.

[Suppurative meningitis in adults due to rare species of the causative agent]. / Gnoinye meningity vzroslykh, vyzvannye redkimi vidami vozbuditelia.

Altogether 20 patients with purulent meningitis caused by agents of rare species were observed. Difficulties in etiological diagnosis and the severity of a clinical course were noted. In most cases this disease developed in persons with associated diseases. In a lingering course of the disease repeated bacteriological investigations of blood and liquor, and serological investigations are recommended.

[Specific latex preparations in the etiological diagnosis of suppurative bacterial meningitis]. / Spetsificheskie lateks-preparaty v étiologicheskoi diagnostike gnoinykh bakterial'nykh meningitov.

The results of the evaluation of the diagnostic latex preparations Bactigen, manufactured by Wampole Laboratories (USA) and intended for the detection of meningococcal antigens, serogropus A, B, C, Y, pneumococcal polyantigens and type b Haemophilus influenzae antigens in the spinal fluid and blood of patients with meningococcal infection and purulent bacterial meningitides, are presented. The pathological material was studied by traditional methods and by the latex agglutination (LAG) test. 522 LAG tests were made, including 414 tests for meningococcal infection, 60 tests for pneumococcal infection and 48 tests for type b H. influenzae. The results of this study revealed that the latex preparations were highly specific with respect to type b H. influenzae antigens and meningococcal antigens (false positive reactions constituted 0.96%). The simplicity of the test and the rapid techniques making it possible to obtain results within 30-40 minutes indicate good prospects of using the LAG test in laboratory practice.

[Etiological diagnosis of suppurative bacterial meningitis today]. / Etiologicheskaia diagnostika gnoinykh bakterial'nykh meningitov v sovremennykh usloviiakh.

As the result of laboratory examination of 2165 patients with virulent bacterial meningitides, including cases of meningococcal infection, the etiological diagnosis was confirmed in 1407 patients (65.0%), the number of cases confirmed by the laboratory examination being significantly greater among adults than among children: 67.5 +/- 1.37% and 63.1 +/- 1.53%, respectively, (t = 2.1). Meningococcal infection was confirmed in 1111 (70.6%) out of 1572 patients under examination. Of the patients with purulent meningitides, pneumococcal etiology was determined in 27.4%, type b of Haemophilus influenzae in 13.5%, other infective agents in 10.0%. The comparison of the results obtained in the examination, carried out by different methods, of 946 children and 770 adults with meningococcal infection revealed a considerable difference in the number of positive results yielded by the bacteriological method and countercurrent immunoelectrophoresis (CIE). Among adults meningococci were isolated twice as frequently (41.1 +/- 2.5% - 19.4 +/- 1.6%), and the results yielded by CIE were predominantly positive (55.1 +/- 2.5% and 40.1 +/- 2.5%). CIE and the immunoenzyme assay were shown to have advantages in the diagnosis of the disease. Under the conditions of intensive antibiotic therapy the methods based on the detection of specific antigens in body fluids can greatly assist in diagnosis.

[Etiology of suppurative bacterial meningitis]. / Etiologiia gnoinykh bakterial'nykh meningitov.

The results of the laboratory examination of 2034 patients with meningococcal infection and purulent meningitides, hospitalized during the period of June 1980 to October 1983, revealed that three main etiological agents were responsible for these diseases: meningococci, pneumococci and Haemophilus influenzae. The susceptibility of the patients to different etiological agents was found to depend on their age. Children aged up to 3 years constituted 75% of the patients with meningitis caused by H. influenzae; 50% of the patients with meningococcal infection were children aged up to 5 years; pneumococcal meningitis occurred more frequently in adults. Serogroup A meningococci were found to prevail in patients with meningococcal infection. Besides, in children serogroup C meningococci could be isolated in 24% of cases. Since 1983 the cases of the isolation of strains belonging to serogroup B increased in number. Among the pneumococci responsible for the disease serotypes 1, 19, 6 and in children serotype 12 occurred most frequently.

[Immunoenzyme method in the diagnosis of meningococcal infections]. / Immunofermentnyi metod v diagnostike meningokokkovoi infektsii.

The materials on the development and use of the test system, based on the enzyme-linked immunosorbent assay (ELISA) and intended for the detection of specific group A and C meningococcal polysaccharides and type b Haemophilus influenzae polysaccharide in the spinal fluid of patients, are presented. In this work commercial preparations manufactured in the USSR were used, and all parameters of the assay were developed on the basis of these preparations. The study was made on the samples of spinal fluid from 410 patients; of these, 203 had meningococcal infection, 57 had purulent bacterial meningitides and 150 had other diseases (acute respiratory diseases, influenza, etc.). As demonstrated by the results of this study, ELISA proved to be a highly specific and sensitive technique. In the investigation of the spinal fluid samples from the patients with meningococcal infection the use of ELISA with bacteriological techniques increased the number of positive results to 67%; with countercurrent electrophoresis, to 78%; and with bacterioscopy, to 83.8%. ELISA is recommended for practical use as an auxiliary laboratory technique and as a rapid method for the diagnosis of meningococcal infection.

[Specific phages used for differentiation of the entomopathogenic bacterium Bacillus thuringiensis]. / O primeneii spetsifichnykh fagov dlia differentsiatsii entomopatogennykh bakterii, otnosiashchikhsia k Bacillus thuringiensis

The sensitivity to specific phages and morphological, physiological, and antigenic properties were compared among several strains of Bacillus thuringiensis isolated from insects inhabiting various geographical zones. All 43 cultures assigned to Bac. thuringiensis var. sotto and 198 among 170 cultures classed as Bac. thuringiensis var. dendrolimus were found to belong to Bac. thuringiensis var. dendrolimus. None of these cultures was resistant to its specific phage. The same was true of 22 studied cultures of Bac. thuringiensis var. galleriae. Only two among studied 45 cultures of Bac. thuringiensis var. thuringiensis were resistant to phages specific for this variety. Therefore, the abundance of variants resistant to specific phages in natural conditions differs among the varieties of Bac. thuringiensis. In most cases, cultures of the same variety of Bac. thuringiensis isolated from various insects inhabiting different geographical zones are identical by their sensitivity to specific phages and by other important characteristics.