Biomedical subjects
Publications and source records attributed to A M Rich.
The purpose of this article is to bring to the attention of clinicians the diagnostic difficulties encountered in some patients with connective tissue syndromes. The relevant literature is reviewed, and a case of undifferentiated connective tissue disease in a patient with an oral mucosal disorder is described.
One hundred and eighteen patients with oral squamous cell carcinoma were staged according to the TNM and STNMP systems. Although both staging systems separated patients into those with a good prognosis and those with a poorer prognosis the more sophisticated system (STNMP) did not provide any additional information. A possible explanation for this anomaly lies in the weighting used in the STNMP system.
The clinical sign of a 'halo-like' white border to an ulcer of the oral mucosa is discussed and the pertinent histopathology described. The term 'peripheral keratosis' has been applied to this appearance which can be useful in the initial clinical assessment of an oral mucosal ulcer.
In an attempt to extend and update information relating to oral cancer in Australia and to provide a basis for international comparison a group of 244 patients suffering from oral cancer have been studied in detail. These patients were all diagnosed in one Oral Pathology Biopsy Service used almost exclusively by oral surgeons and dentists, and although this introduced bias into the sample it had the advantage of standardising documentation. The male: female ratio (1.3 : 1) of patients with oral cancer was lower than that previously reported in Australia, but the average age of patients (60 years) was comparable with many previous reports. The floor of the mouth was the most common site of cancer. There was a significant over-representation of tobacco and alcohol consumers amongst the patients, but a group of elderly females who had never used tobacco or alcohol, yet developed oral cancer, was identified. It was of particular interest that these elderly females had significantly lower haemoglobin and serum iron levels than a sex- and age-matched control group.
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Aggregation of dissociated sponge cells has been proposed as a model for cell-cell recognition mediated by a specific proteoglycan aggregation factor (Microciona aggregation factor). To test whether sponge cells undergo stimulus-response coupling in which intracellular Ca is a messenger, aggregation of mechanically dissociated cells was studied. Changes in light transmission through cell suspensions paralleled aggregation as judged by microscopy. In the presence, but not absence, of Ca (>5 mM) partially purified Microciona aggregation factor aggregated both living and glutaraldehyde-fixed cells. Evidence for a messenger role of Ca was the following: (i) Addition of Ca to Ca-depleted cells induced aggregation that varied with [Ca]. (ii) Addition of Ca ionophores (A23187 and ionomycin) caused aggregation that varied with [Ca] and far exceeded that provoked by Ca alone. Glutaraldehyde-fixed cells did not respond to ionophores with or without Ca. (iii) Calcium antagonists inhibited aggregation. These included inhibitors of the Ca-calmodulin complex (N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride and 1-[bis(p-chlorophenyl)methyl]-3-[2,4-dichloro-beta-(2,4-dichlorobenzyloxyl)phenylethyl]imidazolinium chloride), Ca channel blockers (La, Co, Cd, and verapamil), and three nonsteroidal anti-inflammatory agents (indomethacin, ibuprofen, and piroxicam). Results indicated not only that early events of sponge aggregation can be quantified by continuous recording but that aggregation is not simply due to passive agglutination of inert cells by an extracellular proteoglycan. Rather, sponge cells recognize surface ligands to which they respond by Ca-dependent stimulus-response coupling.
Neutrophils respond to a variety of stimuli by generating superoxide anion, degranulating, and aggregating. Because it has been suggested that fusion of granules with the plasmalemma (degranulation) is necessary for aggregation and superoxide anion generation, we have tested whether these responses can be demonstrated in "neutrophilic cytoplasts" (granule-free vesicles of cytoplasm enclosed by plasmalemma). When examined by electron microscopy, cytoplasts were found to be approximately 4 microns in diameter and essentially granule free. Cytoplasts exposed to fMet-Leu-Phe (0.1 microM) generated superoxide anion after a lag of 16 sec but released no detectable beta-glucuronidase, lysozyme, or elastase. Aggregation of cytoplasts, as measured by changes in light transmission, was also activated by fMet-Leu-Phe; no lag period was observed. Electron microscopy of the aggregates demonstrated clusters of cytoplasts with a scalloped appearance. Superoxide anion generation and aggregation of cytoplasts were also activated by phorbol 12-myristate 13-acetate, concanavalin A, and leukotriene B4. Exposure of cytoplasts to the dye 3,3'-dihexyloxacarbocyanine iodide (DiOC6(3)] led to dye uptake and enhancement of fluorescence, implying that the vesicles were sealed and maintained a membrane potential across the plasmalemma. Exposure of DiOC6(3)-loaded cytoplasts to fMet-Leu-Phe and PMA caused a rapid loss of dye fluorescence that was not inhibited by CN-, compatible with their lack of mitochondria. Exposure of dye-loaded cytoplasts to concanavalin A or leukotriene B4 caused an increase in fluorescence--i.e., a hyperpolarization. These results demonstrate that degranulation is not a prerequisite for aggregation or superoxide anion generation. The retention of ionic gradients and changes in membrane potential, as measured by DiOC6(3) fluorescence changes, suggest a fundamental role for ionic movements in activating superoxide anion generation and aggregation.
Neutroplasts, which are vesicles consisting of cytoplasm enclosed by plasmalemma, have been prepared and found to be incapable of degranulation in response to f-Met-Leu-Phe. However, neutroplasts generate superoxide anion in response to f-Met-Leu-Phe and PMA, and therefore, degranulation is not essential for superoxide anion generation. In addition, neutroplasts aggregate and show shape changes in response to f-Met-Leu-Phe and PMA, and thus degranulation is not essential for aggregation. Neutroplasts take up the carbocyanine dye DiOC6(3), thereby providing evidence for ionic gradients. Dye-loaded neutroplasts show fluorescence changes in response to a variety of stimuli. This response is not CN--inhibitable; therefore, activation of neutroplasts is associated with changes in membrane potential at the plasmalemma, suggesting a role for ion fluxes in the activation sequence for aggregation and superoxide anion generation.
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An abscess that developed following the extraction of periodontally involved teeth persisted after surgical drainage and ampicillin therapy. Subsequent culture of pus from this abscess gave a pure growth of Actinobacillus actinomycetemcomitans which was resistant to ampicillin. Surgical drainage and the use of appropriate antibiotic therapy cleared the infection. The identification of A. actinomycetemcomitans and the types of infection it causes are described. The probable mechanism of infection by the bacterium is discussed. A case that illustrates the importance of the microbiologic examination of pus from dental abscesses is reported.
The random migration of neutrophils under agarose as measured by the number of cells leaving the well, is enhanced when the pH or the osmolality of the medium is reduced or when microtubule agents are used. Concentrations of colchicine above 5 x 10-7 M increased the number of cells migrating and decreased the mean number of centriolar microtubules in a dose-dependent fashion from 16 to 4 per 4 micron 2 at 10-5 M. The distance that colchicine-treated neutrophils migrated from the well was not different from the control. Lowering the pH from 7.4 to 6.0 also increased random migration and decreased pericentriolar microtubules from a mean of 16 to a mean of 10. At pH 6.0, both the number of cells that migrated and the distance the cells forming the leading edge travelled from the well were increased. Since peripheral microtubules may play a greater role in cell migration than centiolar ones, we examined the numerical density of microtubules in the peripheral cytoplasm. Lowering the medium pH reduced the mean number of microtubules per 10 micron 2 from 6 to 2. Colchicine reduced micro-tubules in the same area to I. At the low pH, colchicine reduced even further the numbers of both centriole-associated and peripheral microtubules but the migration pattern was the same as that seen at pH 6.0 without colchicine. Lowering medium osmolality from 280 to 230 m-osmol increased random migration but did not affect microtubule numbers. The addition of colchicine to this system decreased microtubule numbers and increased migration even further. Conditions that enhanced neutrophil migration also affected cell shape. Whereas cells at pH 7.4 were generally fan-shaped with a broad, smooth leading edge, cells at pH 6.0 with or without colchicine were long and narrow. Neutrophils at pH 7.4 but 230 m-osmol had a scalloped edge, which often appeared thickened. This too was not altered by colchicine. The morphology of cells treated with colchicine was similar to controls except for the more frequent presence of long retraction fibres. Each of these treatments thus appears to act on a different aspect of the cell's locomotory apparatus. The mechanisms by which colchicine and lowered ph enhance migration may partially overlap since both significantly decrease peripheral microtubules. The data suggest that microtubules play a constraining role within the cell, limiting the ability of the cell to move and change direction.
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