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Biomedical subjects

L H Browne

Publications and source records attributed to L H Browne.

5 recordsLinked to original sources
PubMed1993

Pure populations of Dictyostelium discoideum prespore and prestalk cells obtained by flow cytometry have different redevelopment characteristics at their cell surfaces.

The multicellular slug stage of Dictyostelium discoideum consists of two major differentiated cell types: prespore and prestalk cells, which become, respectively, the spores and the stalk of the fruiting body. It is known that these cells, although expressing cell-type-specific proteins, remain totipotent, and experimental disruption of slugs results in redifferentiation taking place. We looked at what happens to cell-type-specific surface molecules when a cell changes from one type of another. Using monoclonal antibodies and flow cytometry we were able for the first time to obtain pure populations of single cells of each cell type. These were analysed during redevelopment. The initial hypothesis was that a proportion of each cell type would redifferentiate to reestablish the original proportions. However, it was found that the two cell types responded quite differently. Whereas almost all prestalk cells retained their prestalk surface antigen, in contrast, all prespore cells redifferentiated. During this process redifferentiating prespore cells simultaneously expressed surface determinants of both cell types, an event not seen in normal development.

Animals↗
PubMed1989

Re-expression of 117 antigen, a cell surface glycoprotein of aggregating cells, during terminal differentiation of Dictyostelium discoideum prespore cells.

117 antigen is a glycoprotein expressed on the surface of D. discoideum cells at aggregation. It then disappears and is later re-expressed on the surface of a subpopulation of cells at culmination, the terminal differentiation stage (Sadeghi et al. 1987). A cDNA clone was used to show that the appearance of cell surface 117 antigen accurately reflects the expression of the 117 gene as measured by mRNA levels. It was also shown that during multicellular development there is a reciprocal relationship between the levels of 117 mRNA and the mRNA which codes for prespore surface glycoprotein, PsA. Dual parameter flow cytometry was used to demonstrate that the 117 antigen is found on the surface of maturing prespore cells after the PsA glycoprotein disappears, but that it is not found on mature spores. Using three monoclonal antibodies which identify respectively 117 antigen, PsA, and MUD3 antigen (a spore coat glycoprotein--probably Sp96), two new stages of final spore maturation were defined. These results indicate that there is a recapitulation of at least one aggregative cell surface glycoprotein in the prespore subpopulation of cells as they rise up the stalk during final spore development. This raises the possibility that culmination, which involves complex three dimensional morphogenetic movements not unlike those observed during animal embryogenesis, involves components of the two-dimensional pattern seen during aggregation.

Antigens, Differentiation↗
PubMed1988

Detergent treatment of Dictyostelium discoideum cells allows examination of internal cell type-specific antigens by flow cytometry.

Monoclonal antibodies are used extensively in flow cytometry to identify subpopulations of cells differing in surface antigens. Conventional studies on living cells do not allow analysis of internal antigens, because antibody molecules do not pass through an intact plasma membrane. It is important for developmental studies on Dictyostelium discoideum that not only surface but also internal antigens be analysed. Here techniques are reported that make possible such studies by permeabilising cells with mild detergent treatments using digitonin. Flow cytometer profiles of unfixed cells show that antigens recognised by two monoclonal antibodies, MUD102 and MUD3, are found inside subpopulations of cells in the D. discoideum slug. Double-labelling experiments were carried out to demonstrate that the antigens recognised by these antibodies are present inside prespore but not prestalk cells. The detergent treatment leads to loss of forward-angle light scatter, but 90 degrees light scatter of cells is not greatly affected. While fixed cells sometimes gave satisfactory results, internal labelling did not reliably demonstrate the two subpopulations observed with unfixed cells.

Animals↗
PubMed1987

An evaluation of planned early postnatal transfer home with nursing support.

A community-based programme of planned early postnatal transfer home with the continuity of hospital nursing care was instituted in a defined geographic area of the western suburbs of Sydney in 1983. Mothers were offered the option of discharge in 24-48 hours after delivery, with home visits by a hospital midwife, subject to certain medical and social criteria. An evaluation of the programme in terms of morbidity, psychosocial impact on the family and costs was undertaken. For evaluation, a quasi-experimental study of parallel groups was designed in preference to randomized selection as it was believed that the personal choice would be fundamental to the success of the scheme. A contemporary control group was achieved with volunteer mothers who opted for the traditional five- to seven-day hospital stay. Studies of maternal response and the partner's response and adjustment were undertaken, including the administration of questionnaires that were designed to detect the presence of mild postnatal depression. No increased morbidity occurred in the early discharge group. The early discharge group performed more favourably on the questionnaire that was designed to measure their postpartum adjustment. Continued postnatal domiciliary surveillance reduces the risk that early neonatal pathological changes, especially jaundice, may be overlooked.

Adaptation, Psychological↗