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B C Ross

Publications and source records attributed to B C Ross.

53 records · Page 3Linked to original sources
PubMed1991

Characterization of hepatitis A virus capsid proteins with antisera raised to recombinant antigens.

The capsid proteins of hepatitis A virus (HAV) were expressed as fusion proteins of beta-galactosidase in E. coli using the expression vector lambda gt11. Four fusion proteins were stably expressed and used to immunize rabbits to obtain mono-specific antisera. The antisera were unable to neutralize viral infectivity or react with HAV by radioimmunoassay. Three of the antisera were able to recognize HAV antigens in infected BS-C-1 cells by immunofluorescence and denatured capsid proteins by immunoblot analysis. The antisera were used to investigate the migration of the capsid proteins in gels by immunoblot analysis using standard SDS-PAGE conditions and in gels containing urea. The migration of VP1 and VP3 correlated with their molecular weights predicted from the nucleotide sequence and was consistent in either the presence or absence of urea. However, VP2 migrated with an apparent molecular weight significantly higher than the predicted value and, in gels containing urea, migrated as a doublet. It is proposed that the upper band of this doublet represents VP0, the proteolytic precursor of VP2 and VP4. The relative molecular mass (Mr) of VP4 was estimated to be less than 1 kDa, which is substantially lower than the 2.5 kDa predicted from the nucleotide sequence.

Animals↗
PubMed1991

Differentiation of Mycobacterium tuberculosis strains by use of a nonradioactive Southern blot hybridization method.

The only means of dividing strains of Mycobacterium tuberculosis is by phage typing. Attempts at developing a convenient method based on differences in chromosomal DNA by detecting restriction fragment length polymorphisms (RFLP) have had limited success. This report describes the development of a nonradioactive RFLP technique that differs from most methods by using enzymes that have four base recognition sites rather than six. The restriction enzymes AluI, DdeI, HinfI, NdeII, RsaI, and TaqI were used to digest genomic DNA, and high-molecular-weight fragments were visualized after Southern blotting with digoxigenin-labeled M. tuberculosis DNA. Digestion with AluI resulted in different banding patterns among all eight clinical isolates of M. tuberculosis and three type strains from the tuberculosis complex. NdeII distinguished all but two strains, while HinfI and RsaI were unable to distinguish two pairs of strains. Digestion with the enzymes DdeI and TaqI failed to result in clearly discernible bands. These preliminary results suggest that this method provides a fine differentiation of M. tuberculosis strains and may be useful as an epidemiologic tool.

Blotting, Southern↗
PubMed1991

Identification of Mycobacterium kansasii by DNA hybridization.

A DNA probe specific for Mycobacterium kansasii was obtained from a plasmid clone library of EcoRI-digested genomic DNA. The probe specifically identified culture-confirmed isolates of M. kansasii and isolates in cultures of environmental water samples. In an attempt to distinguish between isolates of M. kansasii, we used two methods to demonstrate restriction fragment length polymorphisms in the genomic DNA. Both of these methods failed to detect any differences between the isolates. These isolates included the type strain TMC 1201, environmental isolates, and clinical isolates from Australia and the Solomon Islands. This result suggests that the genome of M. kansasii is highly conserved and that genetic divergence within this species in insignificant.

Australia↗
PubMed1990

Morphogenesis of hepatitis A virus: isolation and characterization of subviral particles.

The morphogenesis of hepatitis A virus (HAV) in BS-C-1 cells was examined by immunoblotting with antisera to capsid proteins and labeling of virus-specific proteins with L-[35S]methionine. Antiserum to VP2 detected two virus-specific proteins with apparent molecular masses of 30.6 and 30 kDa, representing VP0 and VP2, while antiserum to VP1 detected proteins with molecular masses of 33 and 40 kDa, representing VP1 and a virus-specific protein which we designated PX, respectively. Sedimentation of cell lysates revealed the presence of virions, procapsids, and pentamers, but particles analogous to the protomers of other picornaviruses were not detected. Although provirions and virions were not found as discrete species in our gradient system, it was evident that the rate of sedimentation was proportional to the relative amounts of VP0 and VP2 in particles, with slower-sedimenting particles (provirions) containing predominantly VP0 rather than VP2. Procapsids contained VP0 in addition to VP1 and VP3. Pentamers also contained VP0, but PX was present rather than VP1. These results suggest that PX is a precursor to VP1 and is most likely 1D2A. Primary cleavage of the viral polyprotein also occurs at the 2A-2B junction in cardioviruses and aphthoviruses, but assembly of pentamers containing 1D2A has not been reported for those viruses. The absence of detectable levels of protomers suggests a high efficiency of pentamer formation, which may be related to the high efficiency of viral RNA encapsidation for HAV (D.A. Anderson, B.C. Ross, and S.A. Locarnini, J. Virol. 62:4201-4206, 1988). The results of this study reveal further unusual aspects of the HAV replicative cycle which distinguish it from other picornaviruses and may contribute to its restricted replication in cell culture.

Capsid↗
PubMed1989

Noise exposure of motorcyclists.

This study was designed to evaluate noise exposure of motorcyclists at work. Open and full face safety helmets were compared and the effects of helmet design and the contribution of radio intercoms investigated. Special measuring equipment is described and hearing protection discussed. Previous studies concentrated mainly on the attenuation of noise by safety helmets, and were made under conditions untypical of day-to-day motorcycling. During town driving the sample equivalent continuous noise level (s.leq) ranged from 63 to 90 dB(A), the intercom giving the highest peak levels. On the open road s.leq was up to 105 dB(A); exposure to this level for only 15 min would exceed the present recommended maximum of 90 dB(A) for an 8 h working day.

Head Protective Devices↗
PubMed1989

Nucleotide sequence of high-passage hepatitis A virus strain HM175: comparison with wild-type and cell culture-adapted strains.

The nucleotide sequence of cDNA from a high-passage, cell culture-adapted variant of hepatitis A virus strain HM175 was compared with the previously determined sequences of wild-type virus and two other cell culture-adapted variants. A total of 42 nucleotide changes were detected when the sequence was compared with wild-type virus. Five of these changes were common to all cell culture-adapted strains and a further two changes were shared by the strains that had experienced the greatest number of cell culture passages. The mutations were distributed throughout the genome coding for amino acid substitutions in regions 2B, 2C and 3D with silent changes in 1C and the 5' non-coding region. The possible relevance of these mutations to cell culture adaptation and attenuation is discussed.

Animals↗
PubMed1988

Restricted replication of hepatitis A virus in cell culture: encapsidation of viral RNA depletes the pool of RNA available for replication.

The replication of hepatitis A virus (HAV) in BS-C-1 cells was examined under single-cycle growth conditions by using strand-specific probes for detection of viral RNA species. No measurable lag phase was demonstrated between accumulation of positive-strand HAV RNA and production of infectious virions, indicating that replication of virion RNA is rate limiting for the production of infectious virus. Intracellular viral RNA was further analyzed by using 2 M LiCl to fractionate the insoluble nonvirion 35S RNA and replicative intermediates (RI) from the soluble virions and double-stranded replicative forms, in conjunction with sucrose density gradient ultracentrifugation to separate the different forms of viral RNA. Throughout the productive phase of HAV infection, 95 to 97% of positive-strand HAV RNA was soluble in 2 M LiCl and was shown to be contained in mature virions. Of the LiCl-insoluble HAV RNA, more than 99% was positive-stranded 35S RNA, whereas 0.4% was negative stranded and had the sedimentation and partial RNase resistance characteristics of RI. The pattern of RNA accumulation in HAV-infected cells is thus very different from that seen in poliovirus-infected cells, where large pools of RI and mRNA are produced before RNA is sequestered into mature virions. The results of this study suggest that encapsidation of positive-strand HAV RNA inhibits transcription at all times during the growth cycle, thereby reducing the pool of replicating RNA and the final yield of infectious HAV.

Capsid↗
PubMed1987

Atypical campylobacters associated with gastroenteritis.

Nine strains of Campylobacter species other than Campylobacter jejuni, Campylobacter coli, and Campylobacter laridis were isolated from patients with acute diarrhea. All nine strains showed preferred growth at 37 degrees C under microaerophilic conditions. Conventional microbiological tests and DNA-DNA dot blotting were used to identify these strains. Three of the nine Campylobacter strains hydrolyzed hippurate, reduced nitrate, produced catalase, were resistant to cephalothin, and were shown to be highly related to C. jejuni type strains. Two strains had negative or weak catalase activity and were hippurate negative. Three other strains had characteristics similar to those of Campylobacter cinaedi. The ninth strain, isolated from a homosexual man with antibodies to human immunodeficiency virus (human T-cell lymphotropic virus type III), showed unique features different from those of all the known campylobacters used in this study. This strain grew well at 25 and 37 degrees C and was catalase and nitrate positive, hippurate negative, and resistant to cephalothin.

Adult↗
PubMed1986

Molecular cloning of cDNA from hepatitis A virus strain HM-175 after multiple passages in vivo and in vitro.

Hepatitis A virus (HAV) strain HM-175 was passaged six times in marmosets, 59 times in cell culture and purified from infected cell culture supernatant fluid. The viral RNA was extracted, copied into cDNA and the cDNA:RNA hybrids were cloned into the PstI site of plasmid pBR322. The cDNA clones were authenticated by hybridization to RNA extracted from HAV-infected cells and clones representing the 3' end of the genome were identified using a previously authenticated cDNA clone. The clones represented all but 29 bases of the HAV genome. They were compared to HAV strain HM-175 cDNA cloned from viral RNA after three passages in marmosets on the basis of restriction endonuclease mapping and DNA sequencing. No differences were found in either the presence or absence of restriction endonuclease sites using 33 different restriction enzymes. Sequencing of cDNA representing bases 29 to 1002 of the HAV genome revealed eight base changes all of which were within the 5' noncoding region.

Animals↗
PubMed1985

In vitro histamine H2-antagonist activity of the novel compound HUK 978.

Histamine stimulated adenylate cyclase from guinea-pig fundic mucosa and 3H-tiotidine binding in guinea-pig cerebral cortex were used to assess the in-vitro histamine H2-activity of the novel H2-antagonist HUK 978. The results showed that HUK 978 was a more potent H2-antagonist than either cimetidine or ranitidine. HUK 978 was also shown to be devoid of activity at the histamine H1-receptor, the muscarinic receptor and the alpha and beta-adrenergic receptors.

Adenylyl Cyclases↗
PubMed1985

Anti-secretory activity of the novel H2-antagonist, HUK 978, in rat, guinea-pig and dog.

The anti-secretory activity of the competitive H2-antagonist HUK 978 was determined in rat, guinea-pig and dog. In all systems examined, HUK 978 was more potent than cimetidine and ranitidine both intravenously and orally. In addition, the compound at approximately equipotent doses as these established H2-antagonists exhibited a significantly longer inhibitory profile following oral and systemic administration. Data from these pharmacological studies and the in vitro investigations previously reported, suggest that HUK 978 is a highly specific H2-antagonist and inhibits acid secretion for longer periods than other competitive compounds.

Administration, Oral↗
PubMedDate not supplied

Genetic differentiation of Candida albicans strains by mixed-linker polymerase chain reaction.

Restriction fragment length polymorphisms (RFLPs) of C. albicans were visualized using a mixed-linker polymerase chain reaction (PCR) and a C. albicans-specific primer (C. albicans 1059-bp species-specific repeat sequence). The method produced 10-14 bands of between 200 and 1400 bp on agarose gel electrophoresis and ethidium bromide staining. It gave comparable results to Southern blot hybridization with good reproducibility and the time required for the production of typing profiles was reduced to less than 2 days.

Base Sequence↗