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Biomedical subjects

C Norman

Publications and source records attributed to C Norman.

At least 19 recordsLinked to original sources

U5 snRNA interacts with exon sequences at 5' and 3' splice sites.

U5 snRNA is an essential pre-mRNA splicing factor whose function remains enigmatic. Specific mutations in a conserved single-stranded loop sequence in yeast U5 snRNA can activate cleavage of G1----A mutant pre-mRNAs at aberrant 5' splice sites and facilitate processing of dead-end lariat intermediates to mRNA. Activation of aberrant 5' cleavage sites involves base pairing between U5 snRNA and nucleotides upstream of the cleavage site. Processing of dead-end lariat intermediates to mRNA correlates with base pairing between U5 and the first two bases in exon 2. The loop sequence in U5 snRNA may therefore by intimately involved in the transesterification reactions at 5' and 3' splice sites. This pattern of interactions is strikingly reminiscent of exon recognition events in group II self-splicing introns and is consistent with the notion that U5 snRNA may be related to a specific functional domain from a group II-like self-splicing ancestral intron.

Base Sequence

Mutations in yeast U5 snRNA alter the specificity of 5' splice-site cleavage.

Recognition of 5' splice sites in pre-mRNA splicing is achieved in part by base pairing with U1 snRNA. We have used interactive suppression in the yeast Saccharomyces cerevisiae to look for other factors involved in 5' splice-site recognition. This approach identified an extragenic suppressor that activates a cryptic 5' splice site. The suppressor is a gene for U5 snRNA (snR7) with a single base mutation in a strictly conserved 9 base sequence. This suggests that U5 snRNA can play a part in determining the position of 5' splice-site cleavage. Consistent with this, we have been able to isolate other mutations in the 9 base element in U5 snRNA that specifically activate a second cryptic 5' splice site nearby.

Base Composition

The serological responses of chickens to mass vaccination with a live V4 Newcastle disease virus vaccine in the field and in the laboratory. 1. Meat chickens.

Meat chickens on commercial broiler farms were vaccinated once at 1 to 15 days of age with a live V4 Newcastle disease virus (NDV) vaccine administered by drinking water, aerosol or coarse spray. Hatchmates were housed and similarly vaccinated in laboratory isolation pens. Samples of birds were bled at weekly to fortnightly intervals and the serums tested for haemagglutination inhibiting antibody to NDV. Log2 mean titres of up to 6.26, and assumed protection levels (based on the percentage of birds with log2 titres of 4 or greater) of up to 89%, were obtained in field trials within 4 weeks of vaccination. Differences were observed between the results obtained from parallel field and laboratory trials. The presence of maternal NDV antibody reduced the response to vaccination. The results show that this V4 vaccine can produce an adequate serological response following mass administration to Australian meat chickens housed under commercial conditions.

Aerosols

The serological responses of chickens to mass vaccination with a live V4 Newcastle disease virus vaccine in the field and in the laboratory. 2. Layer pullets.

Layer chickens on a commercial started pullet farm were vaccinated once at 31 to 52 days of age by drinking water or aerosol with live V4 Newcastle disease virus (NDV) vaccine. Flockmates which had been rehoused in laboratory isolation pens shortly beforehand were similarly vaccinated. Samples of birds were bled at intervals and the serums tested for haemagglutination inhibiting antibody to NDV. Log2 mean titres of up to 4.88 and assumed protection levels (based on the percentage of birds with log2 titres of 4 or greater) of up to 81%, were obtained in the field trials within 4 weeks of vaccination. A subsequent laboratory trial further compared the response of different breeds of chicken to different routes of vaccination. Differences were observed between breeds, routes of vaccination, and parallel field and laboratory trials. The results show that this V4 vaccine can produce an adequate serological response following mass vaccination of Australian layer pullets housed under commercial conditions, and that care should be exercised in extrapolating results obtained under laboratory conditions.

Aerosols

Natural infection of broiler breeder chickens with endemic apathogenic Newcastle disease virus and their subsequent response to vaccination with a live V4 Newcastle disease virus vaccine.

Flocks of broiler breeder chickens housed on a commercial farm were monitored from 13 w of age for natural infection with endemic lentogenic Newcastle disease virus (NDV). Seroconversion was first detected at 17 w. By 24 w, all 8 flocks had achieved peak log2 mean haemagglutination inhibiting antibody titres of up to 4.8. Antibody titres then declined and rose again over several months, suggesting cyclic reinfection with NDV. A lentogenic NDV indistinguishable from V4 was isolated from the cloaca of one bird at 18 weeks of age. At 54 weeks of age, 6 of 8 flocks were vaccinated en masse with live V4 NDV vaccine, 3 flocks by drinking water and 3 flocks by aerosol. All flocks were serologically monitored for a further 8 w. Drinking water vaccination induced an anamnestic response in 3 flocks, showing that flocks with pre-existing active immunity to NDV may be successfully vaccinated with V4. However, in all aerosol vaccinated flocks, the procedures failed to induce a response different to that observed in unvaccinated flocks. The serological response to vaccination was greater in sires than in dams.

Animals

The resistance of meat chickens vaccinated by aerosol with a live V4 Newcastle disease virus vaccine in the field to challenge with a velogenic Newcastle disease virus.

Meat chickens housed on a commercial broiler farm in Australia were vaccinated once at 10 to 11 days-of-age by aerosol with live V4 Newcastle disease virus (NDV) vaccine. Groups of vaccinated and unvaccinated birds were flown to Malaysia, where they were challenged with a virulent strain of NDV. Survival rates in vaccinated chickens challenged 7, 14, 21 or 31 d after vaccination were 0.47, 0.77, 0.97 and 0.92, respectively. All unvaccinated chickens died due to Newcastle disease (ND) following challenge. Chickens in Australia and Malaysia were bled and the serums tested for haemagglutination-inhibiting (HI) antibody to NDV. Many vaccinated birds with no detectable antibody, and all birds with a log2 titre of 2 or greater, survived challenge. The results showed that this V4 vaccine induced protective immunity in a significant proportion of chickens within 7 d of mass aerosol vaccination. This early immunity occurred in the absence of detectable circulating HI antibody. Non-HI antibody mediated immunity continued to provide protection up to 31 d after vaccination. Almost all vaccinated birds were protected within 3 w of vaccination. It is concluded that the V4 vaccine is efficacious and could be useful during an outbreak of virulent ND in Australia.

Aerosols

Molecular pathology of chronic myelogenous leukemia.

The presence of Philadelphia chromosome t(9:22) is a hallmark of 95% of clinical cases of chronic myelogenous leukemia (CML) as well as 20% of adult acute lymphoblastic leukemia (ALL) and 5% of acute myeloid leukemia (AML). The product of t(9;22) is a fusion protein BCR-ABL. The fusion proteins of CML, ALL and AML have increased tyrosine kinase activity and show a transforming potential in vitro and in animal models. The shorter p190 protein is associated almost only with ALL and AML, while the protein p210 is present in both chronic phase and blast crisis of CML and also in 50% of Philadelphia-positive (Ph1+) ALL. In CML the transition from chronic phase to blast crisis is usually accompanied by additional genetic events, e.g. additional chromosomal abnormalities, and oncogene activation(s). The detailed understanding of molecular basis of CML, and Ph1+ ALL and AML provides highly sensitive molecular and serological methods to complement classical cytogenetics. The advantages and limitations of these techniques are described and discussed below.

Animals