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Biomedical subjects

P Webster

Publications and source records attributed to P Webster.

At least 73 records · Page 4Linked to original sources
PubMed1995

As good as anyone: antenatal shared care at an inner Sydney hospital.

An exploratory survey design was used to assess satisfaction with antenatal care over a two-month period of women giving birth in an inner Sydney teaching hospital. Patients received obstetric services from private obstetricians, midwives, the hospital outpatient clinic, or 'shared care' between general practitioners and the outpatient clinic or birth centre. Insurance status and demographic information were collected across all groups. Shared care patients gave reasons why they chose that model of antenatal service. Ten per cent of women in the sample received shared care. Shared care patients were equally as satisfied as those in other modes of care in all but one factor--promptness of service (in which private obstetricians received higher ratings). They also judged shared care to have the advantages of being convenient, personal, and culturally appropriate. Significantly more patients in the shared care group were born overseas and they were less likely to hold private insurance. This paper discusses the results of the current study in the context of the Australian literature, explores some issues surrounding satisfaction research, and suggests further research arising from this work.

Australia↗
PubMed1994

Novel infectious particles generated by expression of the vesicular stomatitis virus glycoprotein from a self-replicating RNA.

Self-propagating infectious particles were produced in animal cells transfected with an RNA replicon encoding a single viral structural protein, the vesicular stomatitis virus glycoprotein (VSV-G). The replicon is derived from an alphavirus, Semliki Forest virus (SFV), and encodes the SFV RNA replicase, but none of the SFV structural proteins. After transfection of the replicon into tissue culture cells, expression of G protein spread from small foci throughout the culture. Supernatants from the cells contained infectious, virus-like particles that could be passaged and were neutralized by anti-VSV serum. The majority of the infectious particles were smaller and less dense than either VSV or SFV. Characterization by electron microscopy showed membrane-enveloped vesicles that contained the VSV-G protein. Infectious particles were apparently generated by budding of vesicles containing VSV-G protein and the RNA replicon. These experiments reveal that an enveloped infectious agent can be much simpler than previously thought.

Animals↗
PubMed1994

Brefeldin A sensitivity and resistance in Schizosaccharomyces pombe. Isolation of multiple genes conferring resistance.

The fungal metabolite brefeldin A (BFA) causes the inhibition of protein secretion and the disruption of the structure and function of the Golgi complex in mammalian cells. Here we show that BFA has identical effects in the fission yeast Schizosaccharomyces pombe which normally contains a Golgi complex of stacked cisternae similar to the Golgi complexes in animal cells. After treatment with BFA, secretion was inhibited, Golgi complexes disappeared, and there was an accumulation of endoplasmic reticulum. These results indicate that the effects of BFA in fungi are very similar to those in mammalian cells and provide direct evidence for an effect of BFA on Golgi morphology in fungi. Five spontaneous BFA-resistant mutants were isolated. Genetic analysis showed that the mutations conferring BFA resistance were dominant and in two separate linkage groups. One of the BFA-resistant mutations was found to be allelic to crm1, a gene affecting chromatin structure. All BFA-resistant mutants overexpressed a 20-kDa protein, and the corresponding gene obr1 was isolated and sequenced. However, obr1 overexpression was not sufficient to confer BFA resistance. Plasmids capable of conferring BFA resistance to wild type cells were isolated from libraries constructed from the two BFA-resistant mutants. These plasmids contain six different genes capable of conferring resistance when present in high copy. One of these genes encoded the transcription factor pap1, a homolog of the mammalian AP1 protein. The overexpression of pap1 probably confers BFA resistance indirectly by inducing expression of one or more other proteins. The isolation of several genes conferring BFA resistance suggests several mechanisms are involved.

Amino Acid Sequence↗
PubMed1994

Transient accumulation of new class II MHC molecules in a novel endocytic compartment in B lymphocytes.

Endocytosis of antigen by antigen-presenting cells results in the production of peptides that bind to newly synthesized class II molecules of the major histocompatibility complex. A new population of class II-enriched vesicles has been discovered in B lymphocytes that accumulate internalized antigen but are distinct from endosomes and lysosomes. These vesicles also transiently accumulate newly synthesized class II and class II-peptide complexes and appear to be a compartment specialized for the transport and loading of class II molecules.

Animals↗
PubMed1994

The invariant chain is required for intracellular transport and function of major histocompatibility complex class II molecules.

The major histocompatibility complex (MHC) class II-associated invariant chain (Ii) is thought to act as a chaperone that assists class II during folding, assembly, and transport. To define more precisely the role of Ii chain in regulating class II function, we have investigated in detail the biosynthesis, transport, and intracellular distribution of class II molecules in splenocytes from mice bearing a deletion of the Ii gene. As observed previously, the absence of Ii chain caused significant reduction in both class II-restricted antigen presentation and expression of class II molecules at the cell surface because of the intracellular accumulation of alpha and beta chains. Whereas much of the newly synthesized MHC molecules enter a high molecular weight aggregate characteristic of misfolded proteins, most of the alpha and beta chains form dimers and acquire epitopes characteristic of properly folded complexes. Although the complexes do not bind endogenously processed peptides, class II molecules that reach the surface are competent to bind peptides added to the medium, further demonstrating that at least some of the complexes fold properly. Similar to misfolded proteins, however, the alpha and beta chains are poorly terminally glycosylated, suggesting that they fail to reach the Golgi complex. As demonstrated by double label confocal and electron microscope immunocytochemistry, class II molecules were found in a subcompartment of the endoplasmic reticulum and in a population of small nonlysosomal vesicles possibly corresponding to the intermediate compartment or cis-Golgi network. Thus, although alpha and beta chains can fold and form dimers on their own, the absence of Ii chain causes them to be recognized as "misfolded" and retained in the same compartments as bona fide misfolded proteins.

Animals↗
PubMed1994

Characterization of extreme apical antigens from Toxoplasma gondii.

We have isolated 26 monoclonal antibodies which specifically recognize the extreme apex of Toxoplasma gondii, a protozoan parasite which attaches to and invades host cells via its specialized apical end. The unique apical organelles which define the phylum Apicomplexa are thought to be involved in mechanical and enzymatic aspects of invasion. Immunoblots, immunofluorescence morphology, and immunogold labeling define six classes of apically localized antigens recognized by these antibodies. Three of the classes are detergent-insoluble and localize to the conoid and the cytoplasmic face of the apical membrane, suggesting that they may be part of the parasite's membrane cytoskeleton. The remaining three classes extract with detergent and are associated with internal membrane bounded vesicles (micronemes and the upper necks of rhoptries). One class of micronemal antigens appears to be cell cycle regulated. This antigen localizes to the cytoplasm, especially the perinuclear region, in thin (recently replicated) parasites, but is apical in larger parasites.

Animals↗
PubMed1994

Meta-analytic review of the efficacy of smoking cessation interventions.

A meta-analysis of randomized and controlled evaluations of the efficacy of smoking cessation interventions compared 146 estimates of the difference in abstinence rates between treated and control conditions (effect sizes) from 85 publications. Simple advice to quit and other brief intervention techniques, nicotine chewing gum and behavioural techniques were all found to be significantly better than relevant control conditions in promoting abstinence, although the results were not homogeneous. In five studies of acupuncture compared with control, consistent results were found showing no benefit for acupuncture.

Journal Article↗
PubMed1994

Subcellular localization of CFTR to endosomes in a ductal epithelium.

Plasma membrane chloride transport by the cystic fibrosis transmembrane conductance regulator (CFTR) may be regulated by cellular processes that affect the cycling of CFTR with the plasma membrane. Testing this hypothesis requires cytochemical evidence for the presence of a subcellular compartment of CFTR. In this study, the subcellular distribution of CFTR in a normal epithelial cell population was characterized using immunofluorescence and immunoelectron microscopy. Two anti-CFTR antibodies, raised against different epitopes of the CFTR molecule, specifically labeled the apical pole of striated duct epithelial cells in tissue sections of rat submandibular gland. By use of electron microscopy, the CFTR immunoreactivity was associated with the apical plasma membrane and the membranes of many subapical vesicles. In this preparation, some of the CFTR-labeled vesicles were also labeled with antibodies against transferrin receptor and rab4, two markers of early endosomes and receptor-mediated endocytosis. These observations provide direct cytochemical evidence for the existence of peripherally located CFTR-expressing endosomes and support the hypothesis that membrane recycling may contribute to CFTR function.

Animals↗
PubMed1994

Prevalence of antibodies to Borrelia burgdorferi in Danish deer.

To estimate the prevalence of antibodies to Borrelia burgdorferi in the most common species of Danish deer, blood samples were collected from roe deer (Capreolus capreolus), fallow deer (Dama dama), and red deer (Cervus elaphus). A total of 156 blood samples were collected primarily from hunts in three areas of Denmark from November 1990 to December 1991. Presence of deer IgG antibodies to the Borrelia burgdorferi strain DK ECM 1 were shown by an indirect immunofluorescence assay at serum titre 1:64. Antibodies to spirochaetes were detected in 52% of roe deer, 38% of fallow deer, and 27% of red deer. There were significant differences between the number of seropositive animals in the three areas of Denmark examined, but no differences could be shown for age, sex or season. The high antibody prevalence indicates that deer are exposed to tick-borne Borrelia burgdorferi throughout Denmark.

Age Factors↗
PubMed1993

Stepwise dismantling of adenovirus 2 during entry into cells.

Adenoviruses enter their host cells by receptor-mediated endocytosis and acid-activated penetration from endosomes into the cytosol and deliver their DNA genome into the nucleus. Our results show that incoming adenovirus type 2 particles undergo a stepwise disassembly program necessary to allow progress of the virus in the entry pathway and release of the genome into the nucleus. The fibers are released, the penton base structures dissociated, the proteins connecting the DNA to the inside surface of the capsid degraded or shed, and the capsid-stabilizing minor proteins eliminated. The uncoating process starts immediately upon endocytic uptake with the loss of fibers and ends with the uptake of dissociated hexon proteins and DNA into the nucleus.

Adenoviruses, Human↗
PubMed1993

The flagellar pocket of trypanosomatids.

The surface of the trypanosomatid forms the interface between the parasite and its host, and has evolved to repel a variety of host anti-microbial defences. The flagellar pocket constitutes a highly differentiated region of the trypanosomatid surface that facilitates internalization of host macromolecules, while restricting host access to the exposed, endocytic receptors of the parasite. In this review, Paul Webster and David Russell discuss the ability of this organelle to accumulate efficiently nutrients obtained from the host as a major factor in the success of this group of parasites.

Journal Article↗
PubMed1993

A simple modification to the LKB 7800 series Knifemaker and a balanced-break method to prepare glass knives for cryosectioning.

A novel method of using the LKB 7800 series Knifemaker to produce glass knives using a balanced break is described. The method produces knives of sufficient quality to section aldehyde-fixed, sucrose-cryoprotected, frozen biological material in a cooled cryochamber of an ultramicrotome. The modifications to the Knifemaker are minimal and, if required, the machine can be returned easily to its normal state after use.

Cryoultramicrotomy↗
PubMed1993

The transferrin receptor in African trypanosomes: identification, partial characterization and subcellular localization.

All eukaryotic cells, including African trypanosomes, require iron for growth and division, and this iron is acquired by the receptor-mediated endocytosis of iron-loaded transferrin (diFe(3+)-transferrin). In trypanosomes transferrin (Tf) has been shown to be delivered into lysosomes and may not recycle back to the cell surface as it does in mammalian cells (Grab, D. J., et al., Eur. J. Cell Biol. 59, 398-404 (1992)). Here, we describe for the first time, the characteristics of a Tf-binding protein with receptor-like properties in Trypanosoma brucei brucei. Bloodstream forms of rodent-adapted T. brucei were incubated with [35S]methionine and detergent lysates chromatographed on a Sephacryl S-300 column. Fractions were incubated with anti-Tf serum to immunoprecipitate Tf/Tf-binding protein complexes. On sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) the molecular mass of the major protein in the immunoprecipitate was 88 to 92 kDa. Tf-binding proteins could also be isolated using diferric Tf-Sepharose. The molecular mass of the major Tf-binding protein, as estimated from Sephacryl S-300 column chromatography, in the presence of detergent, was approximately 90 to 100 kDa and 90 kDa with SDS-PAGE. Each 90 kDa Tf-binding protein was able to bind one molecule of diferric Tf. Since monoclonal antibodies to human and bovine Tf receptors failed to react with any trypanosome proteins, antisera were raised against the T. brucei Tf-binding proteins eluted from Tf-Sepharose at low pH. These antibodies recognized a 90 kDa protein on Western blots of a T. brucei lysate and inhibited the growth of T. brucei in vitro. Immunolocalization studies, using this antiserum showed that the Tf-binding protein was localized in the flagellar pocket and within the early endosomal compartments. In the presence of protease inhibitors there was additional localization in lysosome-like organelles. The Tf-binding characteristics and localization of this 90 kDa protein suggest that this molecule is a strong candidate as a physiological receptor for Tf in these parasites.

Animals↗
PubMed1992

Lysosome recruitment and fusion are early events required for trypanosome invasion of mammalian cells.

Trypanosoma cruzi invades most nucleated cells by a mechanism distinct from classical phagocytosis. Although parasites enter at the lysosome-poor peripheral cell margins, lysosomal markers are immediately incorporated into the parasitophorous vacuole. No accumulation of polymerized actin was detected around recently internalized parasites, and disruption of microfilaments significantly facilitated invasion. Lysosomes were observed to aggregate at the sites of trypanosome attachment and to fuse with the vacuole at early stages of its formation. Experimentally induced, microtubule-dependent movement of lysosomes from the perinuclear area to the cell periphery enhanced entry. Conditions that deplete cells of peripheral lysosomes or interfere with lysosomal fusion capacity inhibited invasion. These observations reveal a novel mechanism for cell invasion:recruitment of lysosomes for fusion at the site of parasite internalization.

Animals↗
PubMed1992

The small GTP-binding protein rab4 controls an early sorting event on the endocytic pathway.

rab4 is a ras-like GTP-binding protein that associates with early endosomes in a cell cycle-dependent fashion. To determine its role during endocytosis, we generated stable cell lines that overexpressed mutant or wild-type rab4. By measuring endocytosis, transport to lysosomes, and recycling, we found that overexpression of wild-type rab4 had differential effects on the endocytic pathway. Although initial rates of internalization and degradation were not inhibited, the transfectants exhibited a 3-fold decrease in fluid phase endocytosis as well as an alteration in transferrin receptor (Tfn-R) recycling. Wild-type rab4 caused a redistribution of Tfn-R's from endosomes to the plasma membrane. It also blocked iron discharge by preventing the delivery of Tfn to acidic early endosomes, instead causing Tfn accumulation in a population of nonacidic vesicles and tubules. rab4 thus appears to control the function or formation of endosomes involved in recycling.

Animals↗
PubMed1992

Desialylation of lysosomal membrane glycoproteins by Trypanosoma cruzi: a role for the surface neuraminidase in facilitating parasite entry into the host cell cytoplasm.

Trypanosoma cruzi enters host cells via formation of an acidic vacuole which is subsequently disrupted, allowing the parasite access to the cytoplasm. We show that in an acid environment, release of the parasite surface neuraminidase is enhanced, and this release is likely mediated by a phosphatidylinositol-specific phospholipase C (PIPLC), since antibodies to a carbohydrate epitope (CRD) revealed in glycosylphosphatidylinositol (GPI)-anchored proteins after PIPLC cleavage remove the great majority of the soluble neuraminidase activity from culture supernatants. The neuraminidase is active at acidic pH, and is capable of desialylating known vacuolar constituents, i.e., lysosomal membrane glycoproteins. Parasite escape into the cytoplasm is significantly facilitated in terminal sialylation-defective mutant Lec 2 cells, and enzymatically desialylated membranes are more susceptible to lysis by a parasite hemolysin previously implicated in vacuole membrane rupture. These findings provide evidence that terminal sialylation on carbohydrate moieties contributes to maintaining lysosomal membrane integrity, and indicate a role for a protozoan-derived neuraminidase in facilitating parasite entry into host cells. These observations raise the possibility that other microbial neuraminidases may serve a similar function in acidic intracellular compartments.

Animals↗