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Biomedical subjects

P Whyte

Publications and source records attributed to P Whyte.

At least 37 records · Page 2Linked to original sources
PubMed2001

Distribution and prevalence of airborne microorganisms in three commercial poultry processing plants.

Airborne microbial contaminants and indicator organisms were monitored within three poultry processing plants (plants A, B, and C). In total, 15 cubic feet (c.f.) of air was sampled per location during 15 visits to each plant and quantitatively analyzed for total mesophilic and psychrophilic aerobic counts, thermophilic campylobacters, Escherichia coli, and Enterobacteriaceae. The prevalence of Salmonella spp. in air samples was also evaluated. Significant reductions in total aerobic counts were observed between defeathering and evisceration areas of the three plants (P < 0.05). Mesophilic plate counts were highest in the defeathering areas of all plants compared to equivalent psychrophilic plate counts. Enterobacteriaceae counts were highest in the defeathering areas of all three plants with counts of log10 1.63, 1.53, and 1.18 CFU/15 c.f. recovered in plants A, B, and C, respectively. E. coli enumerated from air samples in the defeathering areas exhibited a similar trend to those obtained for Enterobacteriaceae with log10 1.67, 1.58, and 1.18 CFU for plants A, B, and C, respectively. Thermophilic campylobacters were most frequently isolated from samples in the defeathering areas followed by the evisceration areas. The highest mean counts of the organism were observed in plant A at 21 CFU/15 c.f. sample with plants B and C at 9 and 8 CFU/sample, respectively. With the exception of low levels of Enterobacteriaceae recovered from samples in the on-line air chill in plant A, E. coli, Enterobacteriaceae, or Campylobacter spp. were not isolated from samples in postevisceration sites in any of the plants examined. Salmonella spp. were not recovered from any samples during the course of the investigation.

Air Microbiology↗
PubMed1999

SHIP-mediated inhibition of K562 erythroid differentiation requires an intact catalytic domain and Shc binding site.

Growing evidence supports a role for the SHIP inositol 5'-phosphatase in the negative regulation of a variety of receptor-mediated signaling pathways in hematopoietic cells. SHIP expression among cultured cell lines was examined and found to be restricted to cells of hematopoietic origin, with the exception of the K562 erythroleukemia cell line, in which SHIP protein and mRNA were undetectable. The absence of endogenous SHIP in K562 cells provided a useful system to study the role of SHIP in growth and differentiation. When stably expressed in K562 cells, SHIP was found to be constitutively tyrosine phosphorylated and associated with endogenous Shc and Grb-2. Stable expression of SHIP did not affect growth of the cells but resulted in decreased synthesis of hemoglobin protein and epsilon-globin mRNA in response to hemin, an inducer of erythroid differentiation. This effect was not due to increased cell death in the SHIP-expressing lines following hemin stimulation, but was likely the result of an impaired differentiation program in these cells. Mutational analysis indicated that SHIP must retain both an intact catalytic domain and Shc binding site to efficiently inhibit K562 erythroid differentiation.

Adaptor Proteins, Signal Transducing↗
PubMed1999

A new pathway for mitogen-dependent cdk2 regulation uncovered in p27(Kip1)-deficient cells.

BACKGROUND: The ability of cyclin-dependent kinases (CDKs) to promote cell proliferation is opposed by cyclin-dependent kinase inhibitors (CKIs), proteins that bind tightly to cyclin-CDK complexes and block the phosphorylation of exogenous substrates. Mice with targeted CKI gene deletions have only subtle proliferative abnormalities, however, and cells prepared from these mice seem remarkably normal when grown in vitro. One explanation may be the operation of compensatory pathways that control CDK activity and cell proliferation when normal pathways are inactivated. We have used mice lacking the CKIs p21(Cip1) and p27(Kip1) to investigate this issue, specifically with respect to CDK regulation by mitogens. RESULTS: We show that p27 is the major inhibitor of Cdk2 activity in mitogen-starved wild-type murine embryonic fibroblasts (MEFs). Nevertheless, inactivation of the cyclin E-Cdk2 complex in response to mitogen starvation occurs normally in MEFs that have a homozygous deletion of the p27 gene. Moreover, CDK regulation by mitogens is also not affected by the absence of both p27 and p21. A titratable Cdk2 inhibitor compensates for the absence of both CKIs, and we identify this inhibitor as p130, a protein related to the retinoblastoma gene product Rb. Thus, cyclin E-Cdk2 kinase activity cannot be inhibited by mitogen starvation of MEFs that lack both p27 and p130. In addition, cell types that naturally express low amounts of p130, such as T lymphocytes, are completely dependent on p27 for regulation of the cyclin E-Cdk2 complex by mitogens. CONCLUSIONS: Inhibition of Cdk2 activity in mitogen-starved fibroblasts is usually performed by the CKI p27, and to a minor extent by p21. Remarkably p130, a protein in the Rb family that is not related to either p21 or p27, will directly substitute for the CKIs and restore normal CDK regulation by mitogens in cells lacking both p27 and p21. This compensatory pathway may be important in settings in which CKIs are not expressed at standard levels, as is the case in many human tumors.

Animals↗
PubMed1999

Intact proprioception and control of labour pain during epidural analgesia.

BACKGROUND: Accurate proprioception is critical while walking, yet an ambulatory epidural regimen that provides adequate analgesia for labour while simultaneously preserving proprioceptive input has not been described. METHODS: Sixty primigravidae in established labour received bupivacaine 15 mg (15 ml of 0.1% w/v) and fentanyl 100 micrograms through a lumbar epidural catheter. Clinical assessment of dorsal column sensory function included: vibration sense, distal proprioception and the Romberg test, and were all performed before catheter insertion and 30 min after the study bolus. Sensory modalities were also tested compared to an unblocked dermatome. Pain was scored on a 0-10 cm visual analogue scale (VAS) before and 30 min after induction. Intensity of the motor block was tested using a modified Bromage score (grade 1-6). RESULTS: The study bolus provided reliable analgesia with 43 parturients attaining a VAS pain score of zero. Mean duration of analgesia was 67.5 min (SD 22.85). All parturients retained the ability to perform a partial knee bend while standing (grade 6). No mothers exhibited impaired distal proprioception, altered vibration sense or a positive Romberg sign. CONCLUSION: This study confirms that the addition of lumbar epidural fentanyl 100 micrograms to 15 mg of epidural bupivacaine provides good control of labour pain with no motor block and establishes that this combination preserves dorsal column sensory function.

Adult↗
PubMed1998

Elderly patients with heart failure: a study of satisfaction with care and quality of life.

OBJECTIVE: To assess the level of patient satisfaction with care, patient quality of life, the relationship between these measures and hospital readmission rates, and patient outcomes. METHODOLOGY: A prospective cohort study was conducted of all patients aged 60 years and over admitted to the John Hunter and Mater Hospitals, in the Hunter Area, in whom congestive heart failure contributed to the need for hospital admission. Patients recruited into the main study were then asked to complete a questionnaire on discharge from hospital. Patients were asked to complete the satisfaction questionnaire on discharge because this would reflect the patients' satisfaction with the overall stay. To provide a baseline quality-of-life score, patients were asked to complete a questionnaire 2 weeks after discharge. FINDINGS: Overall scores on the satisfaction questionnaire were high, indicating that the patients were very satisfied with their care. There were six questions in which 17-35% of patients indicated some degree of dissatisfaction, four related to patient knowledge. No difference in satisfaction was found between patients who had or did not have a readmission. Quality-of-life results showed that patients who had a readmission had a significantly lower quality-of-life score at 12 month follow up (P=0.007) than those without a readmission. CONCLUSION: This finding has supported our hypothesis that a higher level of quality of life would be related to fewer readmissions.

Adult↗
PubMed1997

Identification of a p130 domain mediating interactions with cyclin A/cdk 2 and cyclin E/cdk 2 complexes.

P130 shares structural and functional homology with pRb and p107. One property common to p107 and p130, but not to pRb, is the ability to stably interact with cyclin A/cdk2 and cyclin E/cdk2 complexes in vitro and in vivo. Using GST-p130 fusion proteins representing various regions of p130, baculovirus-produced cyclin A/cdk2 and cyclin E/cdk2 complexes were found to interact with residues within a part of p130 known as the spacer region. Cyclin E was able to bind the p130 spacer region in the presence or absence of cdk2 whereas cyclin A binding was dependent upon the presence of cdk2. The smallest p130 fusion protein sufficient to interact with cyclin A/cdk2 or cyclin E/cdk2 complexes contained p130 amino acids 652-698 and deletion of p130 amino acids 680-682 abolished binding to both of the cyclin/cdk2 complexes. When overexpressed in C33A cells, a p130 mutant containing a deletion of amino acids 620-697 was unable to form complexes with either cyclin A or cyclin E. This p130 mutant was at least as active as wild type p130 in suppressing the growth of G418 resistant colonies when overexpressed in C33A or SAOS-2 cells.

CDC2-CDC28 Kinases↗
PubMed1997

The amino-terminal transforming region of simian virus 40 large T and small t antigens functions as a J domain.

Simian virus 40 (SV40) encodes two proteins, large T antigen and small t antigen that contribute to virus-induced tumorigenesis. Both proteins act by targeting key cellular regulatory proteins and altering their function. Known targets of the 708-amino-acid large T antigen include the three members of the retinoblastoma protein family (pRb, p107, and p130), members of the CBP family of transcriptional adapter proteins (cap-binding protein [CBP], p300, and p400), and the tumor suppressor p53. Small t antigen alters the activity of phosphatase pp2A and transactivates the cyclin A promoter. The first 82 amino acids of large T antigen and small t antigen are identical, and genetic experiments suggest that an additional target(s) important for transformation interacts with these sequences. This region contains a motif similar to the J domain, a conserved sequence found in the DnaJ family of molecular chaperones. We show here that mutations within the J domain abrogate the ability of large T antigen to transform mammalian cells. To examine whether a purified 136-amino-acid fragment from the T antigen amino terminus acts as a DnaJ-like chaperone, we investigated whether this fragment stimulates the ATPase activity of two hsc70s and discovered that ATP hydrolysis is stimulated four- to ninefold. In addition, ATPase-defective mutants of full-length T antigen, as well as wild-type small t antigen, stimulated the ATPase activity of hsc70. T antigen derivatives were also able to release an unfolded polypeptide substrate from an hsc70, an activity common to DnaJ chaperones. Because the J domain of T antigen plays essential roles in viral DNA replication, transcriptional control, virion assembly, and tumorigenesis, we conclude that this region may chaperone the rearrangement of multiprotein complexes.

Adenosine Triphosphatases↗
PubMed1995

Characterization of transcription factor E2F complexes during muscle and neuronal differentiation.

The activities of E2F transcription factors are inhibited by interactions with members of the retinoblastoma (RB) tumor suppressor family, p105RB, p107 and p130. In cycling cells p107 and p130 also interact with heterodimers comprised of Cdk2 and either A or E cyclins. We characterized E2F complexes present in C2C12 and P19 mouse cells induced to differentiate into muscle and neuronal cells, respectively. In both undifferentiated C2C12 and P19 cells, in addition to free species, E2F was found in complexes containing p107 or p130 and Cdk2. No E2F-pRB complexes were detected by electrophoretic mobility shift assays even though such cells were shown to contain pRB and E2F species capable of interacting in vitro. These results suggested that although present, pRB was unable to interact with E2F. Following differentiation of C2C12 cells into myotubes, E2F was present in at least two complexes which contained p130, but not in those containing p107 or Cdk2. Low levels of E2F-pRB complexes were also detected in fully differentiated C2C12 myotubes and in freshly isolated skeletal muscle. In the case of differentiated P19 neuronal cells, E2F was found in complexes containing pRB, p107 and p130. However, such cells may not be representative of fully differentiated neurons, as studies with rodent brain extracts indicated that only pRB-E2F complexes and those recognized by a p130-specific serum were present. These results suggested that in both muscle and neurons, pRB and p130 may play specific roles in the development or maintenance of terminal differentiation.

Animals↗
PubMed1995

The retinoblastoma protein and its relatives.

The retinoblastoma tumor suppressor gene is inactivated in retinoblastomas and a variety of other tumor types. In addition, it is one of several cellular proteins targeted by the transforming proteins of the small DNA tumor viruses. At least two other cellular proteins that are targeted by the viral transforming proteins are structurally and functionally related to the protein encoded by the retinoblastoma tumor suppressor gene. Recent data has described a complex series of interactions with cyclins, kinases and transcription factors. It is presumably through these interactions that the retinoblastoma-related proteins exert their effects on cellular proliferation.

Adenovirus E1A Proteins↗
PubMed1993

Diagnosis of Huntington disease: a model for the stages of psychological response based on experience of a predictive testing program.

Persons diagnosed as affected with Huntington's disease (HD) may have similar stages of psychological response to the clinical presentation of the illness. Here we describe a model of these stages of response based on our experience during a predictive testing program for HD. During the Warning Stage, asymptomatic persons are aware of their risk status for HD and develop defenses which favor adaptation to their genetic risk. In response to the initial signs and symptoms of HD (the Incipient Stage) unconscious working through of this realization occurs while it is still kept out of conscious awareness. When symptoms become obvious such that recognition of disease onset is inevitable (Breakthrough Stage) the possibility of the diagnosis of HD is assimilated. After the delivery of the diagnosis during the Adjustment Stage, short- and long-term adaptive responses to living with HD occur. Recognition of the stage of psychological response of a patient who presents with HD is important prior to delivering a clinical diagnosis. In a significant minority of cases, the psychological readiness lags behind the clinical symptomatology and premature presentation of a diagnosis may result in significant untoward adverse events. Understanding of the stages of response may provide a framework for evaluating the psychological state of the person with HD and determining their readiness to receiving the diagnosis. This model may have relevance to the psychological responses of patients to the diagnosis of other late onset autosomal dominant disorders.

Adaptation, Psychological↗
PubMed1993

Conformational constraints of conserved neutralizing epitopes from a major antigenic area of human respiratory syncytial virus fusion glycoprotein.

To study the conformational requirements of epitopes from a conserved antigenic area (area II) of respiratory syncytial (RS) virus fusion (F) glycoprotein, peptides of increasing length containing amino acids essential for these epitopes were synthesized. The synthetic peptides were tested for binding to a panel of neutralizing monoclonal antibodies (MAbs) for this area as well as to rabbit hyperimmune and human convalescent antisera. Antibody binding was dependent on peptide length; thus, a 61-residue peptide spanning amino acids 215 to 275 of the F1 subunit (peptide F215-275) reacted with more antibodies than a shorter (41-residue) peptide F235-275, and this one with more than the (21-residue) peptide F255-275. Most human convalescent sera contained antibodies that reacted with peptides F215-275 and F235-275 but failed to react with F255-275. The results of antibody binding could be related to the structure adopted by the peptides in solution, as determined by circular dichroism spectroscopy and susceptibility of peptides to trypsin digestion. Pretreatment of peptide F215-275 with SDS abolished reactivity with certain MAbs, supporting the notion that higher order structures were needed for antibody binding. High titre anti-peptide antisera were induced in rabbits inoculated with the peptides; however, these sera failed to react with the native F molecule. In mice, only the largest F215-275 peptide induced an anti-peptide response, but their sera reacted poorly with the native F protein and the animals were not protected against an RS virus challenge. These results illustrate the potential use of synthetic peptides in studies of the F protein physical and antigenic structures as well as the problems in designing synthetic RS virus vaccines.

Amino Acid Sequence↗
PubMed1993

The adenovirus E1A-associated 130-kD protein is encoded by a member of the retinoblastoma gene family and physically interacts with cyclins A and E.

The adenovirus E1A proteins form complexes with a group of cellular proteins, including a protein of 130 kD. E1A-associated p130 was purified through coimmunoprecipitation with E1A, and sequence data from four tryptic peptides was obtained. Oligonucleotides derived from the peptide sequences were used to clone a 4.85-kb cDNA. The cDNA contained an 1139-amino-acid open reading frame with homology to the retinoblastoma protein and E1A-associated p107 but was more closely related to p107. In vitro-translated p130 bound to E1A, and anti-p130 antibodies detected p130 in immunoblots of E1A immunoprecipitates. p130 was also detected in immunoprecipitates of cyclins A and E and was an efficient substrate in vitro for kinase activities associated with these cyclins. The p130 gene mapped to chromosome 16q12.2-13, a region that undergoes allelic loss in several types of tumors, including hepatocellular, prostate, and breast carcinomas.

Adenovirus E1A Proteins↗
PubMed1993

Cell cycle-specific association of E2F with the p130 E1A-binding protein.

Association of the E2F transcription factor with the pRb and p107 proteins appears to regulate the activity of E2F and, in turn, affect cell cycle progression. We found, however, that pRb and p107 are only minor E2F-associated proteins in G0/G1 mouse fibroblasts, and we sought to identify the major E2F partner protein in these cells. Because the adenovirus E1A oncoprotein seemed able to bind to the G0 E2F partner, we enriched for proteins that associated both with an E2F-binding site DNA column and with E1A. The major species in G0 and early G1 fibroblasts detected with this approach had properties identical to the pRb- and p107-related p130 protein. In serum-stimulated cells, p107 replaced p130 as the major E2F-associated protein near the G1/S border, concomitant with an increase in p107 protein levels. p130-E2F complexes resembled p107-E2F complexes in their ability to bind to cyclin-cdk kinases, and they appeared to be associated with the cyclin E-cdk2 kinase in late G1 cells. These observations indicate that E2F transcription factors are regulated by a succession of partner proteins with which they associate during defined stages of the cell cycle.

3T3 Cells↗
PubMed1993

Five year study of prenatal testing for Huntington's disease: demand, attitudes, and psychological assessment.

Adult predictive and prenatal testing programmes for Huntington's disease (HD) in Canada have been available since 1986. However, the demand for prenatal testing and the reasons why some people choose not to have the prenatal test for this late onset disorder have not been well documented. In addition, the knowledge and attitudes of adult predictive testing candidates and their partners about prenatal testing are not well known nor are the psychological effects of prenatal testing well understood. As of September 1991, 425 subjects had entered the Canadian Collaborative Study of Predictive Testing and, of these, 47 subjects or their partners had become pregnant. Of this group, 14 (30%) couples requested prenatal testing, 24 (51%) couples did not want prenatal testing, and nine (19%) at risk subjects had already received a decreased risk through adult predictive testing and, therefore, were not eligible for the prenatal test. Of the 14 couples who initially requested prenatal testing, seven withdrew. Thus, demand for the prenatal test by eligible candidates was 7/38 or 18%, which is much lower than the 32 to 65% expected based on early survey data. The most frequently cited reason for declining prenatal testing was the hope that a cure would be found in time for their children. While the majority of adult predictive testing candidates (71%) in our study had accurate information about definitive prenatal testing, many (63%) did not have a correct understanding of exclusion prenatal testing. Although no serious adverse events such as suicide planning or admission to psychiatric hospital have occurred, a particular need for careful counselling was identified for those at risk candidates and their partners who have one prenatal test and feel compelled to use the test again in future pregnancies. Even though prenatal testing for HD is not requested as often originally expected, it still remains a desired option for some at risk persons and their partners.

Adult↗
PubMed1992

The psychological consequences of predictive testing for Huntington's disease. Canadian Collaborative Study of Predictive Testing.

BACKGROUND: Advances in molecular genetics have led to the development of tests that can predict the risk of inheriting the genes for several adult-onset diseases. However, the psychological consequences of such testing are not well understood. METHODS: The 135 participants in the Canadian program of genetic testing to predict the risk of Huntington's disease were followed prospectively in three groups according to their test results: the increased-risk group (37 participants), the decreased-risk group (58 participants), and the group with no change in risk (the no-change group) (40 participants). All the participants received counseling before and after testing. Standard measures of psychological distress (the General Severity Index of the Symptom Check List 90-R), depression (the Beck Depression Inventory), and well-being (the General Well-Being Scale) were administered before genetic testing and again at intervals of 7 to 10 days, 6 months, and 12 months after the participants received their test results. RESULTS: At each follow-up assessment, the decreased-risk group had lower scores for distress than before testing (P < 0.001). The increased-risk group showed no significant change from base line on any follow-up measure, but over the year of study there were small linear declines (P < 0.023) for distress and depression. The no-change group had scores lower than at base line on the index of general well-being at each follow-up (P < or = 0.045). At the 12-month follow-up, both the increased-risk group and the decreased-risk group had lower scores for depression and higher scores for well-being than the no-change group (P < or = 0.049). CONCLUSIONS: Predictive testing for Huntington's disease has potential benefits for the psychological health of persons who receive results that indicate either an increase or a decrease in the risk of inheriting the gene for the disease.

Adult↗
PubMed1992

Characterization of two antigenic sites recognized by neutralizing monoclonal antibodies directed against the fusion glycoprotein of human respiratory syncytial virus.

Two antigenic sites recognized by neutralizing monoclonal antibodies (MAbs) directed against the fusion (F) glycoprotein of human respiratory syncytial virus were mapped on the primary structure of the protein by (i) the identification of amino acid substitutions selected in antibody-escape mutants and (ii) the reactivity of synthetic peptides with MAbs. The first site contained several overlapping epitopes which were located within the trypsin-resistant amino-terminal third of the large F1 subunit. Only one of these epitopes was faithfully reproduced by a short synthetic peptide; the others might require specific local conformations to react with MAbs. The second antigenic site was located in a trypsin-sensitive domain of the F1 subunit towards the carboxy-terminal end of the cysteine-rich region. One of these epitopes was reproduced by synthetic peptides. In addition, mutagenized F protein with a substitution of serine for arginine at position 429 did not bind MAbs to the second site. These results are discussed in terms of F protein structure and the mechanisms of virus neutralization.

Antibodies, Monoclonal↗