Biomedical subjects
Publications and source records attributed to P Sims.
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The FM4-64, a member of the family of fluorescent dyes, has been applied to the cerebellar cortex to evaluate its properties as an intracellular stain and intracortical tracer. Slabs of hamster cerebellum, 1-2 mm thick, were incubated in 10, 30, and 100 microns solutions of FM4-64 in sodium phosphate buffer and observed in a slow scan confocal laser scanning microscope. Mossy and climbing fibers were traced in the cerebellar white and gray substances. They exhibited a high fluorescence signal at the level of the myelin sheath. Mossy fibers were identified in the granular layer by their typical rosette formation and dichotomous bifurcation pattern. Climbing fiber bundles were observed crossing the granular layer and giving collateral branches around Golgi cell bodies. They ascend to the Purkinje cell layer on their way to the molecular layer. Cerebellar macroneurons (Golgi and Purkinje cells) and microneurons (granule, basket, and stellate cells) showed optimal intracellular staining of cell soma, axonal, and dendritic processes. The z-series of stacks of optodigital sections allowed us to explore in depth the cytoarchitectonic arrangement, nerve and glial cell morphology, and the topographic relationship with the afferent fibers.
The intracortical pathway of cerebellar climbing fibers have been traced by means of scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) to study the degree of lateral collateralization of these fibers in the granular Purkinje cell and molecular layers. Samples of teleost fish were processed for conventional and freeze-fracture SEM. Samples of hamster cerebellum were examined by means of CLSM using FM4-64 as an intracellular stain. High resolution in lens SEM of primate cerebellar cortex was carried out using chromium coating. At scanning electron and confocal laser microscopy levels, the climbing fibers appeared at the white matter and granular layer as fine fibers with a typical arborescence or crossing-over branching pattern, whereas the mossy fibers exhibited a characteristic dichotomous bifurcation. At the granular layer, the parent climbing fibers and their tendrils collaterals appeared to be surrounding granule and Golgi cells. At the interface between granule and Purkinje cell layers, the climbing fibers were observed giving off three types of collateral processes: those remaining in the granular layer, others approaching the Purkinje cell bodies, and a third type ascending directly to the molecular layer. At this layer, retrograde collaterals were seen descending to the granular layer. By field emission high-resolution SEM of primate cerebellar cortex, the climbing fiber terminal collaterals were appreciated ending by means of round synaptic knobs upon the spines of secondary and tertiary Purkinje cell dendrites.
The cerebellar basket cells of mice, hamsters, teleost fishes and human have been studied by means of Golgi light microscopy, confocal laser scanning microscopy, scanning and transmission electron microscopy. Golgi light microscopy showed ascending dendrites toward the molecular layer and descending and transverse axonal collaterals contributing to the formation of Purkinje cell pericellular basket. Confocal laser scanning microscopy provided z-series of optodigital sections of ascending basket cell dendrites and descending axonal collaterals participating in the Purkinje cell pericellular nest and the pinceaux. Scanning electronmicrographs displayed the three-dimensional relief of Purkinje pericellular nest formed by basket cell descending and transverse axonal collaterals. Transmission electron microscopy, used as a complementary technique, showed the synaptic contacts formed by basket cell axonal collaterals on Purkinje cell soma and the axosomatic contacts on basket cells by parallel and climbing fiber endings, basket cell axonal terminals and/or Purkinje cell recurrent axonal collaterals. Pre- and postsynaptic membrane specializations were mainly found in parallel fiber axosomatic contacts on basket cells, but not in the Purkinje cell pericellular basket and in other axosomatic contacts on basket cells. These latter findings have been adscribed to a phenomenon of cerebellar synaptic plasticity related with motor learning performance. The correlative microscopy approach demonstrates the potential value of these methodologies for studying the three-dimensional aspect of short intracortical circuits in the central nervous system.
We carried out a prospective study of total lymphocyte counts in 124 adult patients who were diagnosed with HIV (human immunodeficiency virus) infection and/or AIDS (acquired immune deficiency syndrome) and were admitted to Port Moresby General Hospital (PMGH) from January to June 2003. The median and mean values of lymphocyte counts in these patients were found to be 0.7 x 10(9)/l and 0.9 x 10(9)/l, respectively, with a standard deviation of 0.7, both of which counts are significantly lower (p < 0.0001) than those found in members of a control population who were well and HIV-antibody-negative. We found that the lower the total lymphocyte count, the more clinically advanced was the HIV disease state. Haemoglobin values were also significantly lower in these patients. For 35% of these patients, tuberculosis was the principal diagnosis made upon being admitted. An apparent 11% of the patients who had a clinical suspicion of AIDS were HIV-antibody-negative. Total lymphocyte count methods could be used in developing countries that do not have appropriate facilities for CD4 count and viral load assays in order to monitor the patient's disease state and progression towards AIDS.
A study of death and dying was carried out in the settlements of Port Moresby in 2003 and 2004. This descriptive study of 53 patients indicated that they are dying from chronic illnesses, particularly tuberculosis and HIV (human immunodeficiency virus) infection. They are dying slowly over several weeks with minimum care from the health services. The study also showed the problem of violent deaths by guns, by stabbing, by beatings and by police in pursuit of criminals. The lack of a well-coordinated ambulance service leads to preventable deaths in road accidents and in childbirth.
A collaborative study involving 26 laboratories and 5 food types was performed to compare the TECRA Listeria Visual Immunoassay (TLVIA) with standard culture methods. Three foods (lettuce, ice cream, and fish fillets), under the jurisdiction of the U.S. Food and Drug Administration, and 2 foods (cooked chicken and cooked ground turkey), under the jurisdiction of the U.S. Department of Agriculture, were used to determine the effectiveness of the TLVIA. Of the 900 samples tested, 300 were inoculated with low levels (1-5 cells/25 g) of Listeria spp. and 300 were inoculated with high levels of Listeria spp. (10-50 cells/25 g). Method agreement between the conventional culture methods and TLVIA (visual) was 94.7%. Method agreement between the conventional culture methods and TLVIA (reader) was 93.6%. The colorimetric polyclonal enzyme immunoassay (TLVIA) for detection of Listeria in foods has been adopted first action by AOAC INTERNATIONAL.