Non-typhoidal Salmonella rates in febrile children at sites in five Asian countries.

There is increased recognition of non-typhoidal Salmonella (NTS) as a major cause of severe febrile illness in sub-Saharan Africa. However, little is known about community-based incidence of NTS in Asia. In a multicentre, community-based prospective Salmonella surveillance study, we identified a total of six NTS cases: three in Karachi, Pakistan, one in Kolkata, India, and two in North Jakarta, Indonesia. No NTS cases were identified in Hechi, People's Republic of China, and Hue, Viet Nam. Three cases were in children under 3 years, and one case was in a child aged 10 years and one in a child aged 15 years. Only one case was an adult (29 years). The highest incidence of NTS infection was in Karachi (7.2 culture-proven NTS cases per 100,000 person years in age group of 2-15 years). However, in comparison with sub-Saharan Africa, the NTS burden in Asia appears rather limited.

[Detection of tuberculosis in the countries of Central Asia by the electronic monitoring system data].

The basic stages of putting into practice of tuberculosis monitoring systems in the Central Asian Region (CAC) (Uzbekistan, Kazakhstan, Kyrghyz, and Tajikistan) with the assistance of the USA Centers for Disease Control and Prevention, Central Asian Region Programs (CDC/CAR), and the USA Agency for International Development in 2000-2006 are considered. These stages comprised: 1) modification of accounting and reporting forms in accordance with the requirements of the uniform statistical tuberculosis registration system; 2) development, adaptation, and introduction of an electronic tuberculosis monitoring and management system (ETMMS) in the regions and countries of Central Asia; 3) epidemiological analysis of information of tuberculosis monitoring systems, by using the elements of evidence-based medicine. At present, policy electronic tuberculosis monitoring systems entirely cover the areas of three countries of the region. The quality of entries and their processing and analysis is assured by a wide training system set up by the CDC/CAR jointly with the leading national tuberculosis facilities of CAR with the support of the Global Fund for AIDS, Tuberculosis, and Malaria Control. The information obtained by ETMMS has permitted a detail comparative analysis of the structure of tuberculosis at the level of individual regions to identify characteristic groups of areas in the demographic and sociooccupational characteristics of new tuberculosis cases.

Nucleotide sequences in human chromosomal DNA from nonhepatic tissues homologous to the hepatitis B virus genome.

DNA extracted from human nonhepatic tissues (placenta and kidney) have been digested with restriction endonucleases and examined by the Southern procedure with cloned 32P-labelled DNA of hepatitis B virus (HBV). In placental DNAs of women with the history of a hepatitis B infection and in one out of four cases of patients with no known HBV exposure or manifestation, HBV-related chromosomal nucleotide sequences were detected. The integration of HBV-related sequences was observed also in human kidney DNA. Moreover, in the placenta of women who had hepatitis B infection prior to delivery, unusual unintegrated forms of HBV have been found. We conclude that HBV sequences can be found not only in hepatic tissue but also in placental and kidney DNA, both of HBV-exposed and in one case even of a nonexposed patient.

A small open reading frame of the hepatitis delta virus antigenomic RNA encodes a protein that elicits antibodies in some infected patients.

A small open reading frame (ORF) was found in the hepatitis delta virus (HDV) antigenomic RNA encoding a short peptide that shares structural similarity with a region of the hepatitis B virus terminal protein. Analysis of all published HDV genome sequences indicates a high degree of conservation for the small ORF. This ORF is located at the 3'-terminal region of the gene encoding the hepatitis delta antigen (HDAg). We speculated that a peptide encoded by this ORF can be represented as the C-terminal domain of a new protein called HDAg'. This protein contains almost the entire sequence represented in the small form of HDAg and a peptide as an additional 'extension' sequence at the C-terminus. Two long synthetic peptides representing the two different types of peptides encoded by the small ORF were synthesized. These peptides were used for the development of an immunoassay for the detection of antibody to the HDAg' specific domain in sera of patients with HDV infection. Among 162 serum samples analyzed, 13 were found to be positive for an antibody reactive with these synthetic peptides. These antibodies were identified in patients with HDV infections and were not found in patients infected with hepatitis B virus, hepatitis C virus, or non-A,non-B,non-C virus. Thus, these data support the identification and existence of a new antigen encoded by the antigenomic RNA of the HDV.

Injections given in healthcare settings as a major source of acute hepatitis B in Moldova.

BACKGROUND: Reported rates of acute hepatitis B are high in many former Soviet Union republics and modes of transmission are not well defined. METHODS: Two case-control studies were undertaken in Moldova to identify risk factors for acute hepatitis B in people aged 2-15 years (children) and > or =15 years (adults). Serologically confirmed acute hepatitis B cases occurring between 1 January 1994 and 30 August 30 1995, were matched on age, sex, and district of residence to three potential controls who were tested for hepatitis B markers to exclude the immune. Stratified odds ratios (SOR) were calculated using bivariate and multivariate methods. RESULTS: In multivariate analysis, compared with the 175 controls, the 70 adult cases (mean age 25 years, 66% male) were more likely to report receiving injections in the 6 months before illness during a dental visit (SOR = 21; 95% CI: 3.7-120), a hospital visit (SOR = 35; 95% CI: 7.2-170), or a visit to the polyclinic (SOR = 13; 95% CI: 2.4-74). Among children, receiving injections during a hospital visit (SOR = 5.2; 95% CI: 1.2-23) was the only exposure reported significantly more often by the 19 cases (mean age 8 years, 68% male) compared with the 81 controls. CONCLUSION: These results, along with reported unsafe injection practices in Moldova, suggest that injections are a major source of hepatitis B virus transmission and highlight the importance of proper infection-control procedures in preventing transmission of blood-borne infections.

PIP: Two case-control studies were conducted between January 1994 and August 1995 to determine the relative importance of injections and other exposures as a source of acute hepatitis B in Moldova among adults (aged 15 years) and children (aged 2-15 years). Results showed that injections in various health care settings were associated with acute hepatitis B and showed a higher proportion among adults compared with children. Contact with an HBsAg-positive person was also associated with illnesses; however, there was no statistically significant association between acute hepatitis B and other exposures. The risk of HBV transmission following percutaneous exposure is high (at least 30%). Calculation of the population attributable to risk suggests that injections associated with acute hepatitis B cases occurred in adults (52%) and children (21%). Adverse effects of injections may not be apparent in causing chronic infections. Transmission of blood-borne pathogens through unsafe injection practices is a problem increasingly identified worldwide.

Enzyme immunoassay for the detection of antibody to hepatitis E virus based on synthetic peptides.

Five synthetic peptides were prepared based on the nucleotide sequence of open reading frames 2 and 3 encoded in the hepatitis E virus (HEV) genome and were used to develop an enzyme immunoassay (EIA) for the detection of anti-HEV activity in sera. Three different approaches were employed to ascertain the optimal preparation of these peptides as an immunodiagnostic reagent, including (1) a mixture of unconjugated peptides, (2) conjugating individual peptides to bovine serum albumin (BSA) followed by mixing each conjugate at various concentrations, and (3) mixing the peptides before conjugation with BSA to create an artificial antigen complex. The third method was superior in discriminating anti-HEV activity in sera previously tested by Western blot (WB). A frequency distribution of optical density values demonstrated that the peptide-based EIA was able to readily discriminate anti-HEV positive sera from sera devoid of anti-HEV activity. To confirm anti-HEV activity a neutralization test was developed using a mixture of 5 unconjugated peptides. With the exception of sera containing high levels of anti-HEV activity, all sera were neutralized greater than 50%. Strong sera required a higher dilution before a 50% neutralization was achieved. The sensitivity of the WB compared to EIA was 89.5% with and overall concordance of 94.8%. The peptide-EIA was used to determine anti-HEV activity in sera collected from various populations worldwide. In six outbreaks of ET-NANB hepatitis in various geographic regions, anti-HEV activity was demonstrated in 78-100% of cases. The peptide-EIA also detected anti-HEV activity in 14 out of 14 follow-up sera obtained 4-6 months after onset of disease and in 2 of 2 of these patients 5 yr after the acute episode. Anti-HEV activity was found in 8.5% of sera obtain from a healthy population residing in an HEV endemic region and 0.5% in two non-endemic regions (P < 0.001). These data demonstrate that a synthetic peptide-based EIA is sensitive for detecting anti-HEV activity in the sera of patients with acute hepatitis E, convalescents, and among healthy individuals.

Rhesus monkeys infected with hepatitis E virus (HEV) from the former USSR are immune to subsequent challenge with an Indian strain of HEV.

Two rhesus monkeys (M. mullata) of approximately two years of age were inoculated intravenously with a 10% suspension of hepatitis E virus (HEV) positive stool from Kirghistan as evidenced by immuno-electron microscopy. Evidence of HEV infection was demonstrated by rise in serum alanine transaminase (ALT) levels and seroconversion of these monkeys to anti-HEV after 1-1/2 months post-inoculation as evidenced by immunoblot. One year after the primary inoculation, these monkeys were challenged with an Indian strain of HEV. No rise in serum ALT levels was noted during an observation period of 6 months. The same inoculum produced HE in two rhesus monkeys. The results showed that strains from India and Kirghistan were antigenically closely related and rhesus monkeys infected with one strain of virus were immune to another strain.

[Synthesis and antigenic activity of peptides from the ORF3 protein sequence of hepatitis E virus]. / Sintez i antigennaia aktivnost' peptidov iz posledovatel'nosti belka ORF3 virusa gepatita E.

The hepatitis E virus (HEV) is an infection agent (detected recently) responsible for an enterically transmitted non-A, non-B hepatitis [1,2]. Hepatitis E is a big problem in many developing countries, including the Central Asian areas of the former Soviet Union [2]. By cloning followed by sequence analysis of the HEV genome, three open reading frames (ORF) have been identified, among them ORF3 encoding a protein containing 123 amino acid residues, the function of this protein being unknown. Recently [3], one of the immunodominant regions of ORF3 protein was revealed between the 91st and 123rd amino acid residue. The purpose of the present study was a more precise localization of epitopes in the C-terminal portion of HEV ORF3 protein by using synthetic peptides.

[Synthesis and immunological properties of the peptide fragment of the hepatitis B virus envelope preS-protein]. / Sintez i immunologicheskie svoistva peptidnogo fragmenta preS-oblasti belka obolochki virusa gepatita B.

We have synthesized the 24-41 fragment of the preS region of the hepatitis B (subtype ayw) envelope. The peptide was prepared by the solid phase synthesis on perfluoropoly-ethylene polymer grafted with polystyrene. The peptide chain was elongated from C-terminus using pentafluorophenyl- and p-nitrophenyl esters of Boc-amino acids. The peptide was cleaved off the solid phase by HBr in CF3COOH, purified by gel filtration, and, after conjugation with serum albumin (BSA), inoculated into mice. The resultant antibodies were shown to react with the peptide. The blood sera from patients with acute hepatitis B reacted with the peptide-BSA conjugates in the immunoenzymic solid phase assay.

[A comparative study of the diagnostic value of peptides, corresponding to various genomic sequences of the hepatitis delta virus]. / Sravnitel'noe issledovanie diagnosticheskoi tsennosti peptidov, sootvetstvuiushchikh razlichnym genomnym posledovatel'nostiam virusa gepatita del'ta.

A set of 5 peptides from the immunodominant region (65-80 aa) of delta-antigen was prepared by solid phase synthesis. Peptide 71-80 was synthesized in two variants with different amino acid residues in positions 73, 74, 76. Free peptides and their conjugates with bovine serum albumin were tested for antigenicity in ELISA. Correlation between the length of the peptide chain and its antigenic activity was noted. Peptides 65-80 and 69-80 demonstrated positive reaction with all sera from acute HDV patients. Both variants of peptide 71-80 reacted with 83% of positive sera. The smaller peptide 73-80 reacted strongly with one positive serum out of five used.

[Non-A, non-B hepatitis: a new form of serum hepatitis]. / Gepatit "ni A, ni B"--novaia forma syvorotochnogo gepatita.

Forty-seven patients with acute icteric form of "non-A-non-B" hepatitis which was diagnosed on the basis of clinico-epidemiological data, confirmed by the lack of HBsAg in the blood serum of the patients tested by sensitive methods (RIA, ELISA), and by the lack of rise in antibody titers to HBsAg and hepatitis A virus were observed. All the patients had undergone parenteral manipulations including blood transfusions in 34% of the cases. The incubation period was 66.9 +/- 7.07 days. "Non-A, non-B" hepatitis ran a comparatively mild course, with rapid regression of biochemical and clinical signs. Chronic persisting hepatitis developed in 5% of the cases. The above results suggest that "non-A-non-B" hepatitis is a distinct form of serum hepatitis.

[Subtypes of HBsAg of the hepatitis B virus in Western Siberia]. / Subtipy HBsAg virusa gepatita B v Zapadnoi Sibiri.

HBs antigens were subtyped in blood samples of intravenous drug-addicts (IDA) and of donors from as many as 10 cities of Western Siberia by using the immune-enzyme assay with 6 high-specific monoclonal antibodies. Two HBsAg subtypes were found, in a ratio of 3% ayw2: 97% ayw3 (varA and varB), in IDA blood samples from Novosobirsk. Three HBsAg subtypes were found, in a ratio of 57% ayw2: 42% ayw3 (varA and varB): 1% adw2, in the donors' blood samples. The obtained data are sufficient for developing the first national sera panel containing different HBsAg subtypes of hepatitis B virus typical of Russia.

[Clinico-epidemiological characteristics and diagnosis of viral non-A, non-B hepatitis with fecal and oral mechanisms of transmission of the infection]. / Kliniko-épidemiologicheskie osobennosti i diagnostika virusnogo gepatita ni A, ni B s fekal'no-oral'nym mekhanizmom peredachi infektsii.

The paper deals with observations of the patients with non-A-non-B hepatitis (NANBH) transmitted by the fecal-oral mode. The disease was diagnosed by ruling out other similar diseases of the liver, primarily viral A (HA) and B (HB) hepatitides, using clinical and epidemiological data and highly sensitive methods of laboratory diagnosis of HA and HB. Cases of NANBH occurred in one of Central Asia regions in the period of the usual seasonal rise of incidence. The disease was more frequent in adults, running a mild course in most patients, although there were also severe forms with fatal outcomes observed only in pregnant women in the second half of pregnancy. The lethality among the pregnant women was 15.7%. Immune electron microscopy of fecal specimens collected from the patients in the early days of jaundice revealed virus-like particles of 27-30 nm in diameter, morphologically similar to HA virus but forming no immune complexes upon treatment with blood sera containing antibody to HA virus antigen.

[Epidemiological characteristics of non-A, non-B viral hepatitis with a fecal-oral transmission mechanism]. / Epidemiologicheskie osobennosti virusnogo gepatita ni A ni B s fekal'no-oral'nym mekhanizmom peredachi.

The analysis of verified cases of non-A-non-B hepatitis with the fecal-oral mode of transmission which had occurred in one of the districts of the Turkmen SSR, 1984-1985, revealed a this infection: an explosive nature of the incidence, number of epidemiological characteristics of this infection: an explosive nature of the incidence, even distribution in the territory of the district and within one residential area, predominant affection of 15-29-year-old subjects, high mortality among hepatitis-affected pregnant women, insignificant number of secondary cases in the families of index cases. The occurrence of these non-A-non-B hepatitis cases was associated with water. The results of virological and serological studies ruled out the role of hepatitis A and B viruses in the etiology of the acute hepatitis cases occurring in the area. Anti-hepatitis A IgM was detected in the blood in only 3% of the patients in 1984 and in 2% in 1985, exclusively in young patients, and HBsAg in 11% and 9%, respectively. Immune electron microscopy revealed virus-like particles 27-30 nm in diameter in fecal extracts from the patients. The antigen of non-A-non-B hepatitis virus was detected in the first days of the jaundice period in feces of 14% of the patients in 1984 and in 11% in 1985 by an enzyme-immunoassay using a test developed at the Institute of Virology of the USSR Academy of Medical Sciences.

[Electron microscopic research in non-A, non-B viral hepatitis with a fecal-oral mechanism of infection transmission]. / Elektronno-mikroskopicheskie issledovaniia pri virusnom gepatite ni A ni B s fekal'no-oral'nym mekhanizmom peredachi infektsii.

Fecal specimens collected in the early stage of the disease from hepatitis patients classified by serological, clinical, and epidemiological data as non-A-non-B hepatitis were examined by means of cross immune electron microscopy. Extracts of feces from such patients were found to contain full, semi-empty, and empty virus particles without envelopes, 27-30 nm in size, forming immune complexes only in the presence of acute and convalescent sera from patients with this type of hepatitis. The lack of serological crosses with hepatitis A and non-A-non-B viruses isolated in other regions suggests that this virus is antigenically distinct. The final solution requires further studies in which methods of molecular biology of viruses should be used.

[Various physico-chemical properties of non-A, non-B hepatitis virus with a fecal-oral route of transmission and specific diagnosis of the illness]. / Nekotorye fiziko-khimicheskie svoistva virusa gepatita ni A ni B s fekal'no-orallnym mekhanizmom peredachi infektsii i spetsificheskaia diagnostika zabolevaniia.

Observations of 416 patients with and 112 convalescents after non-A-non-B virus hepatitis (HnAnB) with fecal-oral mechanism of transmission were carried out in 1984-1986. An enzyme immunoassay (EIA) was developed for the detection of HnAnB antigen in faeces. The rate of this antigen detection varied from 9.1% to 34.6% in different areas. The HnAnB antigen is present in two areas of cesium chloride density gradient: the maximum at 1.39-1.40 g/cm3 and another peak in the zone of 1.18-1.23 g/cm3. The method of radioimmunoprecipitation followed by SDS-polyacrylamide gel analysis showed the presence of several polypeptides of HnAnB virus. Most clearly, protein 30 K was documented, also polypeptides 91-94 K and 48-57 K were demonstrated. The resistance of nucleic acid of HnAnB virus to RNase was tested. The results indicate that the isolated NA is likely to be DNA. The above data suggest that the agent inducing HnAnB belongs to the group of parvoviruses.

[Improvements in the laboratory diagnosis of hemorrhagic fever with a renal syndrome]. / Usovershenstvovanie laboratornoi diagnostiki gemorragicheskoi likhoradki s pochechnym sindromom.

The results of the employment of direct solid-phase enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) for the detection of antigen and antibody to the virus of hemorrhagic fever with renal syndrome (HFRS) are first presented. Unlike the previously used fluorescent antibody technique (FAT), the immunosorbent methods allow the quantitation of te HFRS antigen, the sensitivity of ELISA and RIA methods for antigen detection being 8-16-fold higher than that of FAT. In antibody determinations, a significant advantage of ELISA and RIA consists in reproducibility of the results due to the standardization of the antigen dose. The observed dynamics of antibody response to HFRS virus is peculiar and somewhat similar to seroconversion in epidemic nephropathia in the Scandinavian countries.

[Specific safety and immunogenicity of a subunit vaccine against hepatitis B]. / Izuchenie spetsificheskoi bezvrednosti i immunogennosti sub"edinichnoi vaktsiny protiv gepatita B.

The results of trials of a subunit vaccine against hepatitis B in volunteers are presented. The active substance of the vaccine was HBsAg isolated from the plasma of asymptomatic antigen carriers. The preparation under study is highly immunogenic, shows poor reactogenicity, and safety in use.

[Further study of viral non-A, non-B hepatitis]. / Dal'neishee izuchenie virusnogo gepatita ni A, ni B.

The paper describes the study of non-A-non-B virus hepatitis with parenteral mechanism of the infection transmission. Immunofluorescence method was used to test 9 liver biopsies from patients with non-A-non-B hepatitis using blood sera from convalescents after this disease. In 6 liver preparations, diffuse fluorescence of hepatocyte cytoplasm was observed. No markers of hepatitis A or hepatitis B were found. In the control group of 17 patients with HB and HA, no non-A-non-B hepatitis antigen was detected. Analysis of the blood sera from the acute period by the ELISA demonstrated the presence of anti-HBs in 28.2% of the patients with non-A-non-B virus hepatitis which corresponds to the frequency of their detection in the normal population. Antibody to HBeAg were found in 59.5%, i. e. significantly more frequently than anti-HBs in non-A-non-B virus hepatitis (p less than 0.03) and anti-HBe in the normal population (15%, p less than 0.01). Experiments of DNA-DNA hybridization demonstrated the homology of HBV DNA and that isolated from the blood serum and liver of patients with non-A-non-B virus hepatitis. The above results suggest that the agent or one of the agents of parenteral non-A-non-B virus hepatitis belongs to the group of Hepadnaviruses.

[The etiological structure of acute viral hepatitis in Tadzhikistan in a period of decreased morbidity]. / Etiologicheskaia struktura ostrykh virusnykh gepatitov v Tadzhikistane v period snizheniia zabolevaemosti.

In the period of reduced incidence of viral hepatitis in Tajikistan, January-December, 1990, 1562 patients with acute viral hepatitis (AVH) were examined in the first days of the jaundice phase (928 children under 14 years and 634 adults) in Dushanbe. Markers of hepatitis A, B, and D (HBsAg, anti-HA IgM, anti-HBc IgM, anti-delta IgM) were determined by enzyme immunoassay. Hepatitis A occurred in 25.8% of the patients with AVH, mostly children of 1-6 years, HB in 22.8%, HD co- and superinfection) in 9.2%. In 42.1% of the patients who had no HA, HB, or HD markers in the blood, non-A, non-B hepatitis (mostly hepatitis E) was diagnosed, mainly in the age groups of 30-39 years (70.7%) and 15-29 years (59.2%). Thus, in Tajikistan hepatitis E occurs not only during outbreaks of this infection but also sporadically.