[Synthesis and biological activity of 2,3-secotriterpene acid mono- and diamides].

Four types of amide (C3; C28; C3-C28) conjugates based on 2,3-seco-18alphaH-oleanane and 2,3-secolupane mono- and dicarboxylic acids were synthesized. The range of diamide derivatives was supplemented with C3-C3' and C28-C28' dicondensed amides with two A-secotriterpene backbones educed by reacting monocarboxylic A-secoacids with biogenic amino acid lysine. Compounds with inhibitory action against herpes virus reproduction (EC50 8.7 and 4.1 McM) were found among the synthesized mono- and diamide derivatives containing an ethyl-beta-alaninate fragment. It has been ascertained that diamide with ethyl-beta-alaninate fragment combines anti-herpes virus properties and anti-HIV activity (EC50 5.1 McM). For active compounds, the maximum non-toxic concentration (MNTC)/EC50 ratios ranges from 9.7 to 40.8. The synthesized amide conjugates do not exhibit any marked cytotoxic effects against human tumor cell lines rabdomiosarcoma RD TE32, A549 lung carcinoma and melanoma MS.

[A new unique HIV-1 recombinant form detected in Belarus].

Republican Research-and-Practical Center for Epidemiology and Microbiology, Ministry of Health of Belarus, Minsk The paper presents data on the molecular genetic characteristics of a new HIV-1 recombinant form. The study has shown that the virus is referred to as HIV-1 subtype B in terms of the gag gene and HIV-1 subtype A in terms of the pol and env genes. At the same time the new isolate is closer, in terms of the gag gene, to the HIV-1 DQ207943 strain isolated in Georgia, in terms of the pol gene, to the HIV-1 AF413987.1 strain isolated in Ukraine and, in terms of the env gene to the HIV-1 AY500393 strain isolated in Russia. Thus, the described new HIV-1 recombinant form has the following structure: BgagApolAenv. The gag, pol, and env gene sequences from the new unique HIV-1 recombinant form have been registered in the international database EMBL/Genbank/DDBJ under accession numbers FR775442.1, FN995656.1, and FR775443.1.

[Molecular epidemiology of genital papillomavirus infection in the Republic of Belarus].

AIM: Determination of epidemiologic features of genital HPV infection in women living in the Republic of Belarus, and detection of phylogenetic features of HPV-16. MATERIALS AND METHODS: 1035 samples of epithelial cervix smears from patients with benign pathology, pre-cancer and cervix cancer were included into the study. 35 samples containing HPV-16 type DNAwere selected for sequencing. HPV L1 gene sequencing (300 n.p.) was performed by using 3100-Avant (Applied Biosystems). Sequencing results analysis was performed by using ClustalX (1.8) and BioEdit. MEGA 4.1 was used for phylogenetic analysis of the fragments obtained and tree construction. RESULTS: HPV-16 type is a dominant genotype that is responsible for the development of cervix cancer, its frequency of occurrence is 56.7% among patients with the specified pathology living in the Republic of Belarus. Predominance of East-Asian and European HPV-16 subtypes that can be attributed to a single evolutionary branch are the territorial features. At the same time detection of HPV-16 subtypes that have 0.003 - 0.007 p-distance between probes may indicate the presence of the several progenitors circulating in the country. A unique HPV-16 subtype was sequenced in the Gomel Region that could be one of the progenitors of HPV-16 circulating on the European continent. CONCLUSION: Further studies performed in the field of molecular epidemiology will allow to give a qualitative characteristic of the epidemic process, determine the direction and time of introduction of the virus into the country, control the circulation of the virus in the population.

[Prevalence of high- and low-risk oncogenic human papillomaviruses in patients with external genital pathology].

The paper presents the results of examining 49 patients with genital papillomas, vulvar and vaginal leukoplakia, dysplasia, and cancer. The findings may suggest that human papillomavirus plays an important role in the development of vulvar and vaginal lesions and reconsider the importance of high- and low-risk oncogenic genotypes in the development of benign neoplasms, precancerous conditions, and malignant tumors of the vulva and vagina.

Multiple genotypes and subtypes of hepatitis B and C viruses in Belarus: similarities with Russia and western European influences.

The Republic of Belarus reports a seroprevalence of 4.8% for hepatitis B virus (HBV) and 1.26% for hepatitis C virus (HCV), but little is known about the molecular characteristics of the circulating viruses. This study analysed 69 HBV surface antigen (HBsAg)-positive and 113 anti-HCV-positive donors attending a national reference hospital in Minsk. Among the HCV patients, 70% were co-infected with human immunodeficiency virus (HIV). Phylogenetic analysis of 12 complete genomes and 31 partial HBV sequences, as well as 78 core/E1 HCV sequences, revealed that multiple genotypes and subtypes of both viruses were circulating in Belarus. Of the HBV strains, 11.6% were genotype A2 and 88.6% were genotype D. The genotype D strains segregated into four recently described subtypes, with D2 being the most prevalent (58.1%), followed by D3 (16.3%), D1 (11.6%) and D4 (2.3%), but with inter-subtypic distances lower than the minimal 4% distance proposed to define subtypes. The 78 HCV strains belonged to subtypes 1b (53.8%), 3a (38.5%), 1a (5.1%), 4a (1.3%) and 4d (1.3%). Subtype 1b was less prevalent (45.1% vs. 70.4%) among HCV/HIV co-infected donors, while subtype 3a was more prevalent (29.6% vs. 43.1%). The relative prevalence of HBV and HCV genotypes in Belarus corresponded to the prevalence in Russia, although with a clear European influence that reflected the socio-political context of the country.

[The molecular epidemiology of HIV-1 in Belarus in (1996-2004): a predominance of subtype A variants and circulation of subtype B variants].

To study the molecular epidemiology of HIV-1 in Belarus, the genetic sequences of HIV-1 variants were obtained from 50 infected persons, which represented the main stages, risk groups, and geographic areas of the epidemic. The env and gag sequences were studied for HIV-1 variants from 31 persons, the env sequences were for HIV-1 variants from 18 persons, and the gag sequence was for HIV-1 variant from 1 person. Phylogenetic analysis indicated that the sequences of HIV-1 variants from 46 persons were homogenic and evolutionally closely related to IDU-A strains specific for other epidemics in the former Soviet Union are dominating in the epidemic in Belarus. Circulation of epidemiologically unrelated subtype B viruses was also established.

[Synthesis and antiviral activity of amides and conjugates of betulonic acid with amino acids]. / Sintez i protivovirusnaia aktivnost' amidov i kon''iugatov betulonovoi kisloty s aminokislotami.

Betulonic acid amides with aliphatic and heterocyclic amines and with L-amino acids were synthesized by the acid chloride method. Betulonic acid amide and L-methionine derivatives of betulonic acid and its 3-oxime effectively inhibit the influenza A virus. Betulonic acid octadecylamide is active against the herpes simplex type 1 virus. The conjugate of betulonic acid 3-oxime with L-methionine is also active toward HIV-1. The tested compounds mainly show no activity toward the ECHO6 virus, which is devoid of a coat. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 1; see also http://www.maik.ru.

[Synthesis and antiviral activity of hydrazides and substituted benzalhydrazides of betulinic acid and its derivatives]. / Sintez i protivovirusnaia aktivnost' gidrazidov i zameshchennykh benzal'gidrazidov betulinovoi kisloty i ee proizvodnykh.

New nitrogen-containing derivatives of betulinic and betulonic acids, hydrazides and N'-benzalhydrazides, were synthesized. Their antiviral activities toward of influenza A virus, herpes simplex type I virus, enterovirus ECHO6, and HIV-1 were studied in vitro. Betulinic acid 3-oxime was found to have the highest activity against the influenza virus. Betulonic acid, betulinic acid 4-chlorobenzalhydrazide, betulonic acid 3-oxime benzalhydrazide, and betulinic acid hydrazide inhibited the replication of herpes simplex type I virus. Betulinic acid hydrazide also showed antiviral activity toward HIV-1. All the derivatives of betulinic acid under study displayed a low antiviral activity toward enterovirus ECHO6.

Viral water contamination as the cause of aseptic meningitis outbreak in Belarus.

In the recent years Echovirus-30 associated outbreaks have taken place in different European countries. Aseptic meningitis caused by Echovirus-30 was the main diagnosis of a large outbreak in Belarus in Summer-Autumn, 1997, involving 460 patients. Echovirus-30 was detected in cerebrospinal fluid of the patients with aseptic meningitis. This serotype played the dominant role in the outbreak. Minor serotypes and mixtures of enteroviruses were detected in faeces and nasopharyngeal lavages. Investigation of environmental samples gave evidence of expressed viral contamination of drinking water and water sources (river and ground sources). River water sources were considerably contaminated with viruses. The incidence of virus isolation was 50%. After cleaning procedures, the incidence became two times lower, proving imperfect water purification and disinfection procedures. Sequence analysis of isolates from Belarus (isolates from water and patient's cerebrospinal fluid) showed the difference of 0.2%. The outbreak peculiarities such as high attack rate and wide-spread of the disease incidences, clinical form variability, isolation of outbreak strain from water and a good agreement between minor serotypes isolated from faeces and water samples as well as correlation in the dynamics of acute intestine infections, aseptic meningitis morbidity and bacterial water contamination can be considered as evidence of its water-borne. Echovirus-30 isolates from Belarus were very closely related to each other and to several European isolates. Sequence difference between isolates of 1994-1998 from European countries was found to be 4.3%. The data can point to the common primary source of enterovirus infection, connected to water and to the possibility of epidemic strain transmission from neighbouring states to the Republic of Belarus.

[Water-borne outbreak of serous meningitis caused by Echovirus-30 in Belarus]. / Vodnaia vspyshka seroznogo meningita, vyzvannaia virosom Echo-30, v Belarus.

In recent years outbreak of enterovirus infections caused by Echovirus-30 were rather frequently registered in different European countries. A major outbreak caused by this virus took place during the summer-autumn period of 1997 in the city of Gomel, Belarus. Sanitary epidemiological and molecular epidemiological studies made it possible to determine that the outbreak was water-borne. The sequence analysis of Echovirus-30 strains isolated from water and the cerebrospinal fluid of patients revealed a minor divergence between them (0.2%) indicative of their practical identity. The comparison of the Belorussian isolates with the strains isolated in Europe in 1994-1998 also showed a small percentage of differences in their genomes, which showed that the outbreak of Echovirus-30 infection was probably brought to Belarus from the territories of European countries.

[Increase in the serotypical heterogeneity in HIV-1 in Svetlogorsk focus 1 year after the start of an epidemic]. / Narastanie serotipicheskoi geterogennosti VICh-1 v svetlogorskom ochage cherez 1 god posle nachala épidemii.

Serotypical heterogeneity of HIV-1 variants in the Svetiogorsk focus of HIV-1 epidemic (Gomel region, Belarus) increased in the cohort of intravenous drug addicts in comparison with the situation one year before. A more complex serotypical profile of the epidemic can reflect the routes of virus transmission.

[Use of rapid methods in detection of enteroviruses]. / Ispol'zovanie ékspress-metodov dlia detektsii énterovirusov.

Indirect immunofluorescence (IIF) and RT-PCR were used for rapid diagnosis of enterovirus infection during an outbreak of aseptic meningitis in Gomel. IgM to enterovirus were detected in 93.6% sera by IIF and in 71.4% cerebrospinal fluid specimens by RT-PCR. IIF takes only 1.5-2 h and RT-PCR 6-7 h. The methods are recommended for rapid diagnosis of enterovirus infection.

Dehydrogenase activity of infected cells and biological properties of HIV-1 variants.

Changes in dehydrogenase activity of some continuous cell lines were studied during the response to acute infection in vitro with HIV-1 variants having different biological features. Soon after infection, the dehydrogenase activity of infected cells increased, and this increase was greater and more prolonged with the HIV-1 r/h variant than with the HIV-1 s/l variant. Later stages showed decreased dehydrogenase activity of HIV-1-infected cells compared to the noninfected control; this is a manifestation of the cytodestructive effect of the virus. The changes increased monotonously (but not in direct proportion) with an increase in the infecting dose.

[Serological properties of HIV-1 isolates from the focus of an epidemic in the Gomel region of the Belarus Republic (1996)]. / Serologicheskie svoistva izoliatov VICh-1 iz ochaga épidemii v Gomel'skoi oblasti Respubliki Belarus' (1996 g.).

Immunoreactivity of sera obtained in 1996 from HIV-1-infected intravenous narcomaniacs from an epidemic focus in Gomel region, Belarus, is studied with a panel of 10-16-component synthetic peptides simulating apical epitope of surface glycoprotein gp120 V3 loop of HIV-1 variants. Comparative analysis of resultant spectra with representative immunoreactivity spectra of sera from a sampling of HIV-1-positive sera collected in 1986-1997 in different regions of the former USSR demonstrated a high homogeneity of immunoreactivity spectra of sera from Gomel region. Serotypes of HIV-1 A/C variants circulating in populations of intravenous narcomaniacs in the focus in Gomel region in 1996 and in Southern and South-Eastern Ukraine in 1995-1996 are similar. This confirms a previous conclusion about the prevalence of A/C serotype among intravenous narcomaniacs in the former USSR.

[Phenotypic characteristics of HIV isolates, isolated in the Belarus Republic]. / Fenotipicheskaia kharakteristika izoliatov VICh, vyiavlennykh na territorii respubliki Belarus'.

Biological characteristics of 26 HIV strains circulating in the Republic of Belarus were studied in cell cultures. By the replicative activity, infectivity, and tropism to certain cell lines, all the strains can be divided into 2 groups: rapid/high and slow/low. Slow/low HIV strains were isolated from asymptomatic patients with lymphadenopathy and from AIDS patients. On the other hand, two rapid/high strains were isolated from patients without clinical signs of AIDS 6-8 months after probable infection.

Antibody to hepatitis E virus in HIV-infected individuals and AIDS patients.

Antibody to hepatitis E virus of IgG class (anti-HEV IgG) is regularly detected in industrialized countries, where HEV is non-endemic, at levels not exceeding 2-3%; seropositive individuals are often found in certain groups of patients and professionals exposed to an increased risk of blood-borne infections. The present study was aimed at the identification of anti-HEV IgG in patients with human immunodeficiency virus (HIV) infection, including acquired immune deficiency syndrome (AIDS), living in Russia and Belarus, an area of low anti-HEV prevalence with a moderate spread of HIV infection and AIDS. In Russia, 13 out of 117 HIV-infected patients (11.1%) were found to be anti-HEV seropositive. This differed significantly from the frequency observed in the normal population (1.7%) but not from the frequency in a matching control, high-risk group consisting of male prisoners (8.0%). No difference in the frequency of anti-HEV IgG seropositivity was found between groups of HIV-infected men subdivided by sexual orientation. The rate of anti-HEV seropositivity increased with the progression of HIV infection, reaching 43.3% in AIDS patients and 38.1% in those who died from AIDS. In Belarus, anti-HEV IgG seropositivity was not found among 20 HIV-infected subjects nor among individuals from the control risk group, which consisted of 25 intravenous drug users. In conclusion, HEV infection may have common transmission mechanisms (risk factors) with HIV infection rather than represent an additional opportunistic infection in AIDS.