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T Humphrey

Publications and source records attributed to T Humphrey.

At least 19 recordsLinked to original sources
PubMed2005

A case-control study of domestic kitchen microbiology and sporadic Salmonella infection.

The microbiology of domestic kitchens in the homes of subjects who had suffered sporadic Salmonella infection (cases) was compared with control domestic kitchens. Case and control dishcloths and refrigerator swabs were examined for the presence of Salmonella spp., total Enterobacteriaceae counts and total aerobic colony counts. Salmonella spp. were isolated from both case and control dishcloths and refrigerators but there were no significant differences between the two groups. Colony counts were similar in case and control dishcloths and refrigerator swabs. There was no relationship between the total counts and presence of Salmonella . There was no evidence that cases of Salmonella infection were more likely to have kitchens which were contaminated with these bacteria or have higher bacterial counts than controls. Total bacterial counts were poor indicators of Salmonella contamination of the domestic kitchen environment. Further factors which could not be identified by a study of this design may increase risk of Salmonella food poisoning. These factors may include individual susceptibility of the patient. Alternatively, sporadic cases of Salmonella food poisoning may arise from food prepared outside the home.

Adolescent↗
PubMed2003

Comparative genome analysis of Campylobacter jejuni using whole genome DNA microarrays.

Whole genome DNA microarrays were constructed and used to investigate genomic diversity in 18 Campylobacter jejuni strains from diverse sources. New algorithms were developed that dynamically determine the boundary between the conserved and variable genes. Seven hypervariable plasticity regions (PR) were identified in the genome (PR1 to PR7) containing 136 genes (50%) of the variable gene pool. When comparisons were made with the sequenced strain NCTC11168, the number of absent or divergent genes ranged from 2.6% (40 genes) to 10.2% (163) and in total 16.3% (269) of the genes were variable. PR1 contains genes important in the utilisation of alternative electron acceptors for respiration and may confer a selective advantage to strains in restricted oxygen environments. PR2, 3 and 7 contain many outer membrane and periplasmic proteins and hypothetical proteins of unknown function that might be linked to phenotypic variation and adaptation to different ecological niches. PR4, 5 and 6 contain genes involved in the production and modification of antigenic surface structures.

Algorithms↗
PubMed2003

The microbiological quality of washing-up water and the environment in domestic and commercial kitchens.

AIMS: To determine the microbiological quality of washing-up water and the environment in domestic and commercial kitchens. METHODS AND RESULTS: Chicken meals were prepared by people without food safety training in their own kitchen (n = 52) or by trained staff in a commercial kitchen (n = 10). Study participants then washed-up, cleaned the kitchen and completed a food hygiene questionnaire. The temperature and microbiological quality of the washing-up water, and the presence of pathogens in dishcloths, tea towels and other kitchen samples was determined. Of the raw chickens used in meal preparation, 96 and 13% were naturally contaminated with Campylobacter or Salmonella spp., respectively. In domestic kitchens, two of 45 sponges, dishcloths or scourers and one of 32 hand- or tea towels were contaminated with Campylobacter after washing-up and cleaning but none of the tap or sink swabs yielded pathogens. The mean washing-up water temperature in the domestic kitchens was 40.7 degrees C, whereas in the commercial kitchen it was 44.7 degrees C (P = 0.04). Study participants who used hotter water (>/=40 degrees C) had lower levels of bacteria in their washing-up water. The aerobic plate counts of the washing-up water samples in domestic homes were usually between 105 and 106 CFU ml-1 but those associated with the commercial kitchen were consistently lower (P = 0.01). Despite this, Campylobacter was detected in one of 10 washing-up water samples from the commercial kitchen but in none of the samples from domestic kitchens. CONCLUSIONS: Pathogenic microorganisms can be recovered relatively frequently from the kitchen environment. SIGNIFICANCE AND IMPACT OF STUDY: By identifying factors that affect the number of microorganisms in washing-up water and the kitchen environment, evidence-based recommendations on implementing domestic food hygiene can be made.

Animals↗
PubMed2002

Risk factors for salmonella food poisoning in the domestic kitchen--a case control study.

Domestic kitchen food handling risk factors for sporadic salmonella food poisoning are largely unknown. We compared food consumption and food handling practices, opportunities for cross contamination and refrigerator temperature control, in 99 households in South East Wales in 1997/8 with a case of salmonella food poisoning, and control households matched for electoral ward. On univariate analyses, cases were significantly more likely than control respondents to have purchased free-range eggs in the preceding week, and more likely than control households to have handled frozen whole chicken in the previous week, and to handle raw chicken portions at least weekly. In multivariate analysis, only consumption of raw eggs and handling free-range eggs were significant risk factors, independent of the age structure of the family and of the season.

Adolescent↗
PubMed2001

Changes in the carriage of Campylobacter strains by poultry carcasses during processing in abattoirs.

The recent development of simple, rapid genotyping techniques for Campylobacter species has enabled investigation of the determinative epidemiology of these organisms in a variety of situations. In this study we have used the technique of fla typing (PCR-restriction fragment length polymorphism analysis of the flaA and flaB genes) to identify the sources of strains contaminating the carcasses of five campylobacter-positive and two campylobacter-negative broiler flocks during abattoir processing. The results confirmed that, in the United Kingdom, individual broiler flocks are colonized by a limited number of subtypes of Campylobacter jejuni or C. coli. In some but not all cases, the same subtypes, isolated from the ceca, contaminated the end product as observed in carcass washes. However, the culture methodology, i.e, use of direct plating or enrichment, affected this subtype distribution. Moreover, the number of isolates analyzed per sample was limited. fla typing also indicated that some campylobacter subtypes survive poultry processing better than others. The extent of resistance to the environmental stresses during processing varied between strains. The more robust subtypes appeared to contaminate the abattoir environment, surviving through carcass chilling, and even carrying over onto subsequent flocks. From these studies it is confirmed that some campylobacter-negative flocks reach the abattoir but the carcasses from such flocks are rapidly contaminated by various campylobacter subtypes during processing. However, only some of these contaminating subtypes appeared to survive processing. The sources of this contamination are not clear, but in both negative flocks, campylobacters of the same subtypes as those recovered from the carcasses were isolated from the crates used to transport the birds. In one case, this crate contamination was shown to be present before the birds were loaded.

Abattoirs↗
PubMed2000

A fission yeast homolog of Int-6, the mammalian oncoprotein and eIF3 subunit, induces drug resistance when overexpressed.

Through a screen to identify genes that induce multi-drug resistance when overexpressed, we have identified a fission yeast homolog of Int-6, a component of the human translation initiation factor eIF3. Disruption of the murine Int-6 gene by mouse mammary tumor virus (MMTV) has been implicated previously in tumorigenesis, although the underlying mechanism is not yet understood. Fission yeast Int6 was shown to interact with other presumptive components of eIF3 in vivo, and was present in size fractions consistent with its incorporation into a 43S translation preinitiation complex. Drug resistance induced by Int6 overexpression was dependent on the AP-1 transcription factor Pap1, and was associated with increased abundance of Pap1-responsive mRNAs, but not with Pap1 relocalization. Fission yeast cells lacking the int6 gene grew slowly. This growth retardation could be corrected by the expression of full length Int6 of fission yeast or human origin, or by a C-terminal fragment of the fission yeast protein that also conferred drug resistance, but not by truncated human Int-6 proteins corresponding to the predicted products of MMTV-disrupted murine alleles. Studies in fission yeast may therefore help to explain the ways in which Int-6 function can be perturbed during MMTV-induced mammary tumorigenesis.

Amino Acid Sequence↗
PubMed1998

Cell-division-cycle defects associated with fission yeast pre-mRNA splicing mutants.

We have isolated six new pre-mRNA splicing mutants (prp) from a collection of temperature-sensitive (ts-) Schizosaccharomyces pombe strains. The prp mutants are defective in the splicing of both messenger RNA and U6 small nuclear RNA precursors. A single recessive mutation is responsible for both the ts- growth and the splicing phenotypes in each of the prp mutants. The six prp mutations are unlinked and fall into separate complementation groups. Two are allelic with the previously described prp3 and prp4 mutations; the remaining four define the new alleles prp5-1, prp6-1, prp7-1, and prp9-1. The six mutants exhibit three splicing phenotypes: accumulation of unspliced precursor at the restrictive but not at the permissive temperature; accumulation of unspliced precursor at both the permissive and restrictive temperatures; and accumulation of unspliced precursor, the intron-exon lariat intermediate, and the intron lariat final product. In addition to their aberrant splicing phenotypes, the prp5-1 and prp6-1 mutants express classical cell-division-cycle defects, while prp7-1 exhibits an unusual hyphal morphology. These results suggest a connection between pre-mRNA splicing and the control of cell division in fission yeast.

Cell Cycle↗
PubMed1998

Sum1, a highly conserved WD-repeat protein, suppresses S-M checkpoint mutants and inhibits the osmotic stress cell cycle response in fission yeast.

The S-M checkpoint ensures that entry into mitosis is dependent on completion of DNA replication. In the fission yeast Schizosaccharomyces pombe, the SM checkpoint mutant cdc2-3w is thought to be defective in receiving the checkpoint signal. To isolate genes that function in the checkpoint pathway, we screened an S. pombe cDNA library for genes that, when overexpressed, could suppress the checkpoint defect of cdc2-3w. Using this approach, we have identified a novel gene, sum1+ (suppressor of uncontrolled mitosis). sum1+ encodes a highly conserved WD-transducin repeat protein with striking sequence similarity to the human transforming growth factor (TGF)-beta-receptor interacting protein TRIP-1 and to the translation initiation factor 3 subunit eIF3-p39, encoded by the TIF34 gene in Saccharomyces cerevisiae. S. pombe sum1+ is an essential gene, required for normal cell growth and division. In addition to restoring checkpoint control, overexpression of sum1+ inhibits the normal cell cycle response to osmotic stress. Furthermore, we demonstrate that inactivation of the stress-activated MAP kinase pathway, required for cell cycle stress response, restores the S-M checkpoint in cdc2-3w cells. These results suggest that Suml interacts with the stress-activated MAP kinase pathway and raise the possibility that environmental conditions may influence the checkpoint response in fission yeast.

Amino Acid Sequence↗
PubMed1998

Suppressors of cdc25p overexpression identify two pathways that influence the G2/M checkpoint in fission yeast.

Checkpoints maintain the order of cell-cycle events. At G2/M, a checkpoint blocks mitosis in response to damaged or unreplicated DNA. There are significant differences in the checkpoint responses to damaged DNA and unreplicated DNA, although many of the same genes are involved in both responses. To identify new genes that function specifically in the DNA replication checkpoint pathway, we searched for high-copy suppressors of overproducer of Cdc25p (OPcdc25(+)), which lacks a DNA replication checkpoint. Two classes of suppressors were isolated. One class includes a new gene encoding a putative DEAD box helicase, suppressor of uncontrolled mitosis (sum3(+)). This gene negatively regulates the cell-cycle response to stress when overexpressed and restores the checkpoint response by a mechanism that is independent of Cdc2p tyrosine phosphorylation. The second class includes chk1(+) and the two Schizosaccharomyces pombe 14-3-3 genes, rad24(+) and rad25(+), which appear to suppress the checkpoint defect by inhibiting Cdc25p. We show that rad24Delta mutants are defective in the checkpoint response to the DNA replication inhibitor hydroxyurea at 37 degrees and that cds1Delta rad24Delta mutants, like cds1Delta chk1Delta mutants, are entirely checkpoint deficient at 29 degrees. These results suggest that chk1(+) and rad24(+) may function redundantly with cds1(+) in the checkpoint response to unreplicated DNA.

Amino Acid Sequence↗
PubMed1995

Cell-cycle checkpoints. Keeping mitosis in check.

Mutations in an essential yeast gene, encoding DNA polymerase epsilon, abolish the dependence of mitosis on the completion of DNA replication, suggesting that the replication complex provides the checkpoint signal.

Animals↗
PubMed1995

The isolation of Campylobacter jejuni from contaminated surfaces and its survival in diluents.

The isolation rates of campylobacters from contaminated surfaces were improved if swabs were placed directly into selective media rather than being stored in diluents before culture. Storage in diluents resulted in a loss of viability and the remaining viable campylobacter cells were often sub-lethally injured which sensitised them to selective agents in culture media and reduced isolation rates. Campylobacter jejuni, suspended in small drops of blood, was capable of prolonged survival on work surfaces if the drops remained liquid but the bacterium died rapidly once drops had dried.

Campylobacter jejuni↗
PubMed1994

RNA 3' end signals of the S.pombe ura4 gene comprise a site determining and efficiency element.

We have defined sequences in the 3' non-coding region of the Schizosaccharomyces pombe ura4 gene that are required for efficient mRNA 3' end formation. Three separate sequence elements have been identified. Two of these are site determining elements which specify alternative sites of polyadenylation [the major poly(A) site and a minor downstream poly(A) site]. The third sequence, located downstream of both poly(A) sites, functions as an efficiency element that enhances utilization of either polyadenylation site. By employing sensitive RT-PCR analysis, we demonstrate that although low levels of transcripts are detected up to the efficiency element, none is detected beyond this point. The downstream site determining element and efficiency element have both been delineated to specific 16 nt sequences which we show are together sufficient for ura4 mRNA 3' end formation. We have further characterized the interaction between these two elements and show that the efficiency element behaves in a position-independent, orientation-dependent manner, but cannot form 3' ends independently of the site determining element. Surprisingly, we find that the efficiency element can be functionally replaced by a second copy of either site determining element. We present a model for the mechanism of RNA 3' end formation of the ura4 gene and note that this bipartite structure for a poly(A) signal in S.pombe may be related to the AAUAAA and downstream GU-rich sequences of poly(A) signals in mammalian genes.

Base Sequence↗
PubMed1991

Homologous mRNA 3' end formation in fission and budding yeast.

Sequences resembling polyadenylation signals of higher eukaryotes are present downstream of the Schizosaccharomyces pombe ura4+ and cdc10+ coding regions and function in HeLa cells. However, these and other mammalian polyadenylation signals are inactive in S. pombe. Instead, we find that polyadenylation signals of the CYC1 gene of budding yeast Saccharomyces cerevisiae function accurately and efficiently in fission yeast. Furthermore, a 38 bp deletion which renders this RNA processing signal non-functional in S. cerevisiae has the equivalent effect in S. pombe. We demonstrate that synthetic pre-mRNAs encoding polyadenylation sites of S. pombe genes are accurately cleaved and polyadenylated in whole cell extracts of S. cerevisiae. Finally, as is the case in S. cerevisiae, DNA sequences encoding regions proximal to the S. pombe mRNA 3' ends are found to be extremely AT rich; however, no general sequence motif can be found. We conclude that although fission yeast has many genetic features in common with higher eukaryotes, mRNA 3' end formation is significantly different and appears to be formed by an RNA processing mechanism homologous to that of budding yeast. Since fission and budding yeast are evolutionarily divergent, this lower eukaryotic mechanism of mRNA 3' end formation may be generally conserved.

Base Sequence↗