Vascular endothelial growth factor expression in heart of rats exposed to hypobaric hypoxia: differential response between mRNA and protein.

This study investigated the role of vascular endothelial growth factor (VEGF) in the neoangiogenesis induced in heart in response to hypoxia. The time-course of adaptive changes in capillary supply, expression of VEGF mRNA and protein was studied in right (RV) and left ventricles (LV) of rats exposed to hypobaric hypoxia during 1-25 days (barometric pressure = 505 hPa). VEGF mRNA levels encoding for VEGF 188 and 164 isoforms were measured by reverse transcription-polymerase chain reaction (RT-PCR) and VEGF protein was determined by Western blotting.

Short report: phylogenetically distinct hepatitis E viruses in Pakistan.

Hepatitis E, which is enterically transmitted, is the most common cause of acute hepatitis in much of Asia. Phylogenetic analysis of several isolates of hepatitis E virus (HEV) from Asia suggests that transmission of this virus is geographically restricted. In Sarghoda, Pakistan, HEV Sar-55 was isolated from a 1987 outbreak.

Identification of a novel hepatitis E virus in Nigeria.

Sporadic cases of acute hepatitis E among ten native Nigerian adults were reported in Port-Harcourt (Nigeria). Hepatitis E virus (HEV) was detected in serum and/or faecal samples of seven patients by RT-PCR of the open reading frame (ORF)-1 polymerase region and the 3'-end of ORF2. Restriction analysis widely used to distinguish genotypes I and III showed that all Nigerian strains have a pattern similar to the Mexican strain (NotI, nt 286; SmaI, nt 397; no KpnI restriction site) but displayed a BsmI restriction site at nt 213 as do most African HEV strains sequenced so far.

[Geographical distribution of hepatitis E virus genotypes].

Hepatitis E virus (HEV) is the major agent of acute hepatitis in developing countries where the infection occurs sporadically or in large waterborne epidemics. HEV, classified in the Caliciviridae, is not culturable. The detection of HEV RNA by RT-PCR in serum and stool samples is reliable during the 7 to 15 days following the onset of the disease.

Phylogenetic analysis of hepatitis E virus isolates from Egypt.

Hepatitis E virus (HEV) genome was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in fecal samples of two sporadic cases of hepatitis E in Cairo Egypt. Sequence of the complete putative structural region [open reading frame (ORF)-2] and complete region of unknown function (ORF-3) was determined for the two HEV isolates. Phylogenetic analysis of the nucleotide sequences was performed using neighbor joining or maximum parsimony methods of tree reconstruction.

Experimental African HEV infection in cynomolgus macaques (Macaca fascicularis).

Experimental infection with hepatitis E virus (HEV) from Africa has not been investigated. Our purpose was to study hepatitis E produced by HEV from Chad (North Africa) and to analyze the genetic sequence of the HEV obtained after animal passage. An HEV-containing fecal sample from Chad was intravenously inoculated in four cynomolgus macaques.

Role of hepatitis E virus in sporadic cases of acute and fulminant hepatitis in an endemic area (Chad).

Forty-one patients with acute or fulminant hepatitis and 86 control patients were entered into a study of sporadic, acute, and fulminant hepatitis in the N'Djamena area of Chad in 1993. Acute hepatitis B was diagnosed in nine (22%) patients and acute hepatitis E in 27 (66%) patients. No acute hepatitis A was observed and 10% of the patients had serologic markers of hepatitis C virus (HCV) infection.

Characterization of hepatitis E virus (HEV) from Algeria and Chad by partial genome sequence.

The purpose of this study was to analyze partial nucleotide sequences and derived peptide sequences of hepatitis E virus (HEV) from two outbreaks of hepatitis E in Africa (Chad 1983-1984; Algeria 1978-1980). A portion of ORF3 and the major portion of ORF2 were amplified by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). The PCR products were sequenced directly or after cloning into the pCRII vector.

Short report: polymerase chain reaction detection of hepatitis E virus in north African fecal samples.

Epidemics of enterically-transmitted non-A, non-B hepatitis were described in 1983-1984 involving French soldiers in Chad and in 1979-1980 in residents of Algeria. Hepatitis E virus (HEV) was subsequently implicated by serology. In this study, the presence of HEV in patient stool specimens from both outbreaks and from sporadic cases in residents of Chad (1994) was documented.

Use of digoxigenin-labeled RNA probe to test hepatitis A virus antiviral drugs.

A nucleic acid hybridization assay was used to evaluate inhibitory activity of antiviral compounds against hepatitis A virus (HAV) in cell culture and compared to radioimmunoassay by analysis of variance procedure. The 5' genomic end of the HM-175 strain was used as digoxigenin-labeled RNA probe. Dot-blot examination showed a reduction of detectable HAV RNA in infected cells when treated with amphotericin B.

Studies on mechanism of action of glycyrrhizin against hepatitis A virus replication in vitro.

Glycyrrhizin (GL) achieved a concentration-dependent inhibition of the replication of hepatitis A virus (HAV) in PLC/PRF/5 cells. GL has been shown to inhibit an early stage of the HAV replication. GL was not virucidal and had no measurable effect on the adsorption of [3H]uridine-labelled virions to cells.

Development of an RNA/RNA hybridization assay for the detection of the HAV CF53 strain.

A quick and sensitive dot-blot assay using non-radioactive labelled RNA probes was developed for the detection of the CF53 strain of hepatitis A virus (HAV) in cell culture. The cDNA of the 5' end of the HM175 strain was inserted in a transcription vector pSPT18 and was used to synthesize 32P- or digoxigenin-labelled RNA probes. These RNA probes specifically detected the RNA of the CF53 strain and can be used to detect HAV in PLC/PRF/5 cells.

[Detection of hepatitis A virus by riboprobe labeled with digoxigenin: comparison of methods].

A riboprobe (RNA probe), corresponding to the 5' end of the HM175 hepatitis A virus (HAV) genome, was synthetized in vitro and was digoxigenin-labeled. Then the riboprobe was used to detect the CF53 HAV strain. Conditions of virus denaturation (with or without SDS and proteinase K, timing of assay) to release viral RNA were tested by dot-blot hybridization on a ten fold dilution of HAV suspension.

[Water and viral hepatitis].

The main agents responsible for acute viral hepatitis throughout the world are the hepatitis A virus (HAV) and the hepatitis E virus (HEV). Both are transmitted by fecal-oral route and can provoke large epidemics, HAV in developed countries and HEV in developing countries. Water is a major vehicle of spread.

Antiviral activity of carrageenan on hepatitis A virus replication in cell culture.

Sulphated polysaccharides such as iota-, lambda- and kappa-carrageenans showed a potent inhibitory effect on the replication of hepatitis A virus (HAV) in the human hepatoma cell line PLC/PRF/5. No cytotoxic effects were detected with concentrations of carrageenans up to 200 micrograms/ml. The selectivity indices of these substances, calculated as the ratio of the dose that reduced the number of viable cells to 50% (CD50) to the effective dose that inhibited 50% of viral antigen expression (ED50), were greater than 400 with iota-carrageenan, greater than 222 with lambda-carrageenan and greater than 10 with kappa-carrageenan.

Inhibition of hepatitis A virus replication in vitro by antiviral compounds.

Forty antiviral compounds were screened for inhibitory effect on hepatitis A virus (HAV) antigen expression in the human hepatoma cell line PLC/PRF/5. Ribavirin, amantadine, glycyrrhizin, and pyrazofurin were selected in this screening test and were studied further. The selectivity indices of these four compounds, calculated as the ratio of 50% cytotoxic dose (determined by the trypan blue exclusion and by inhibition of [3H] leucine incorporation) to the 50% effective dose (determined by the viral antigen expression), were 4.

[Comparative study of plasmid resistance to mercury of 2 bacterial strains of animal origin].

Bacterial resistance to mercury has been studied in two different strains from animal origin, Salmonella typhimurium 9205 and Escherichia coli 467. These two strains are resistant to mercuric chloride but sensitive to phenylmercury, and thus belong to the group of bacteria that possess a "narrow" spectrum resistance. The presence of plasmids within the cells has been demonstrated through conjugation experiments and direct detection of extrachromosomal DNA in transconjugants.

[Plasmid resistance to antiseptics. Demonstration of methods].

Susceptibility to chlorhexidine, benzalkonium chloride and mercury chloride was studied for 70 Gram negative strains (56 Enterobacteriaceae and 14 Pseudomonas aeruginosa) recovered from humans or animals. Minimal inhibitory concentration (MIC) was determined on solid agar (replica plating) and in a liquid medium (laser nephelometry). For resistant strains, plasmidic mediation was looked for using conjugation.