Biomedical subjects
Publications and source records attributed to R C Wong.
A case of polyclonal B-cell pharyngeal lymphoma in a renal transplant patient is reported. No evidence of Epstein-Barr virus involvement was detected, but the lymphoma underwent complete remission with acyclovir and reduction in immunosuppression.
CD43 is a constitutively phosphorylated 115-kDa sialoglycoprotein expressed on a variety of blood cells including lymphocytes and monocytes. L10, a mAb directed against CD43, triggers T cell activation and enhances hydrogen peroxide production in monocytes. Activation of mononuclear cells by L10 initiates phosphoinositides hydrolysis, C2+ mobilization, and protein kinase C (PKC) activation. In turn, activated PKC hyperphosphorylates CD43, suggesting a potential role for PKC in the regulation of signaling via CD43. To address this issue, we have analyzed the effect of PKC activation by the tumor promoter PMA on L10-triggered rise in intracellular free Ca2+ concentrations ([Ca2+]i). Treatment of mononuclear cells with PMA profoundly inhibited the increase in [Ca2+]i induced by L10. The inhibition of CD43-mediated signaling by PMA was due, in part, to uncoupling of CD43 from the signal-transducing G protein. This was evidenced by the comparatively modest inhibition by PMA of the increase in [Ca2+]i induced by the direct G protein activator AlF4-. PMA treatment did not affect the surface expression of CD43. However, it induced the hyperphosphorylation of CD43, the extent of which correlated with the inhibition of CD43-mediated increase in [Ca2+]i. Staurosporine, a potent inhibitor of PKC, abrogated the hyperphosphorylation of CD43 and normalized CD43-mediated signaling in PMA-treated cells. Significantly, in the absence of PMA, staurosporine enhanced the rise in [Ca2+]i triggered by L10, suggesting that engagement of CD43 by activating ligands results in feedback inhibition by PKC. It is concluded that activation of PKC inhibits signaling via CD43 by mechanisms involving phosphorylation and uncoupling of CD43 from the signal-transducing apparatus and by distal, post-receptor events.
In order to study the relationship between assembly, surface expression, and signal transduction of the alpha/beta T-cell antigen receptor-CD3 complex (TCR.CD3), a series of T-cell mutants with a partial block in assembly of the complex was generated. By chemical mutagenesis, we produced somatic cell variants of the human T-leukemia cell line, HPB-ALL, which expressed low amounts of TCR.CD3 complexes on their surface. RNA and protein analyses demonstrated that most variants synthesized normal amounts of the individual members of the complex, i.e. TCR-alpha, TCR-beta, CD3-gamma, -delta, -epsilon, and -zeta. In these variants, less than 10% of the TCR.CD3 complexes inside the cell contained the CD3-zeta 2 homodimer due to an intrinsic deficiency in the formation of the TCR-alpha/beta heterodimer. The low level of assembly of CD3-zeta 2 into the TCR.CD3 complex and an additional decrease in the rate of export of the TCR.CD3 complex from the endoplasmic reticulum explained the low level of expression of alpha/beta receptors on the surface of these mutants. Only cells with the complete set of subunits of the TCR.CD3 complex on their surface were capable of transducing CD3-mediated signals. The results presented in this paper indicate that TCR-alpha/beta heterodimer formation is an obligatory requirement for assemblage of CD3-zeta 2 into a functionally competent TCR.CD3 complex.
CD43 (sialophorin, gpL115) is a sialoglycoprotein expressed on a wide variety of blood cells including lymphocytes, monocytes, neutrophils, and platelets. L10, an anti-CD43 mAb, has been shown to induce monocyte-dependent activation and proliferation of human T lymphocytes. We have studied the signaling mechanism involved in this activation process. Treatment of PBMC and purified populations of T cells and monocytes with L10 induced the hydrolysis of phosphoinositides with the resultant generation of the phosphoinositide-derived second messengers diacylglycerol and inositol phosphates. This was associated with the translocation of protein kinase C from cytosol to membrane fractions and an increase in free intracellular Ca2+ in treated cells. In human leukemic T cell lines, the magnitude of signaling via CD43 did not correlate with the density of the TCR/CD3 surface expression nor with the intensity of signaling via the TCR/CD3. Moreover, a mutant derived from the leukemic T cell line HPB-ALL that was severely defective in TCR/CD3 surface expression and signaling nevertheless had normal CD43 surface expression and signaling compared with the parent cell line. It is concluded that CD43 is functionally coupled to the phospholipase C/phosphoinositides signaling pathway. In human T cells, signaling via CD43 proceeds independently of TCR/CD3. The widespread expression of CD43 suggests a potentially important role for this molecule in orchestrating the activation of multiple cell types.
Lymphocyte proliferation is associated with cell-cell aggregation. In order to assess the importance of cell-cell contact in T-cell proliferation we examined the effect of disruption of cellular aggregation by anti LFA-1(4) mAb on T-cell proliferation. Monocyte-dependent T-cell proliferation induced by anti-CD3 mAb, pairs of anti-CD2 mAbs, or PHA was inhibited by anti-LFA-1 mAb. Monocyte-independent proliferation of highly purified T cells to anti-CD3 mAb plus PMA or plus IL-2 and to PHA plus IL-2 was, surprisingly, also inhibited by anti-LFA-1 mAb. Anti-LFA-1 mAb caused the partial inhibition of both low-affinity and high-affinity IL-2 receptor and the complete inhibition of IL-2 synthesis. In contrast to the above, the proliferation of highly purified T cells to PMA plus ionomycin was not inhibited by anti-LFA-1 mAb. These results suggest that optimal activation of highly purified T cells via cell surface receptors requires LFA-1-dependent cell-to-cell contact between proliferating T cells as well as between T cells and accessory cells. Such contact appears to be crucial for initiating IL-2 production and for optimal action of IL-2 through its receptor.
Conventional ventilators are frequently used at high rates in the intensive care nursery to achieve adequate oxygenation and ventilation with reduced peak inspiratory pressure. The efficacy and limitations of high frequency positive-pressure ventilation (HFPPV) using a conventional ventilator were studied by comparing the cardiopulmonary effects of HFPPV with those of high frequency jet ventilation (HFJV) in an animal model of respiratory failure. Sixteen saline-lavaged rabbits were ventilated with either HFPPV or HFJV for 2 h using rates of 200 breaths/min, inspiratory to expiratory ratio of 1:2, and FIO2 of 1.0. As controls an additional eight lavaged rabbits were ventilated at conventional rates (40 to 60 breaths/min). Proximal peak inspiratory pressure as indicated on the ventilator manometer or drive pressure was adjusted to maintain acceptable blood gases. Cardiac output (CO) was measured by thermodilution. Although there was a significant decrease in cardiac function over time, there were no significant differences between the groups in CO or stroke volume. Satisfactory oxygenation and ventilation were maintained in all groups. Static respiratory system compliance and mean airway pressure were similar among the groups. Histologic examination of the lungs revealed no differences between the three ventilator groups. The results of this study indicate that both HFPPV and HFJV are effective in short-term maintenance of normal blood gases in respiratory failure without any discernable differences in their effects on cardiovascular function. At very high rates, however, increases in VT are not possible with HFPPV, which limits its usefulness and flexibility in respiratory failure.
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Phenytoin (diphenylhydantoin; Dilantin) is a highly effective and widely prescribed anticonvulsant and antiarrhythmic agent. Since 1938 it has been invaluable in the treatment of grand mal and psychomotor epilepsy. Hydantoin derivatives have been used medicinally for more than a half-century. In recent years dermatologists have broadened the indications for phenytoin use to include recessive dystrophic epidermolysis bullosa, linear scleroderma, and pachyonychia congenita. In spite of widespread use and popularity, it is interesting that the frequency of complications relating to drug therapy remains low, relatively speaking. Nevertheless, a broad spectrum of cutaneous and immunologic reactions to phenytoin have been reported. These range from tissue proliferative syndromes (side effects), drug hypersensitivity syndromes (allergic effects), and a possible linkage with lymphoma (idiosyncratic effects). Therapeutic and toxic reactions to this commonly prescribed drug are comprehensively reviewed, analyzed, and summarized in this monograph.
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A rapid and sensitive spectrophotometric assay for ornithine decarboxylase is described. It is based on the observation that the product of ornithine decarboxylase, putrescine, reacts with 2,4,6-trinitrobenzenesulfonic acid to give a colored product soluble in 1-pentanol whereas ornithine does not. The amount of putrescine produced by the enzyme was determined by measuring the absorbance of the 1-pentanol extract of the reaction mixture at 420 nm, and by comparing the results to those obtained by the trapping of 14CO2 and by HPLC assays. The three assays were found to be equivalent in sensitivity, with the spectrophotometric assay having the advantages of being relatively rapid, requiring only common laboratory equipment, and not requiring the use of radioactive isotopes.
Spiders are a ubiquitous component of the fauna of North America. At least 50 to 60 species in the United States are known to bite humans, although in most cases, the diagnosis is never suspected nor is treatment necessary. A few species of spiders are capable of causing necrotic wounds and, occasionally, death. This review is intended to provide an overview of the recent advances in the diagnosis and treatment of spider bites, as well as a compilation of the many species of spiders that have been reported to give clinically important bites.
The efficiency of immunoprecipitation and immunoblotting for detecting Trypanosoma cruzi antigens using rabbit and mouse antisera has been examined. Comparison of the starting material for each technique showed that the extraction methods resulted in a similar range of polypeptides when assessed by apparent Mr. Reaction with either hyperimmune rabbit sera or a sequential series of sera from infected mice, showed a significant disparity in the range of antigens revealed by each technique. Epitopes on polypeptides greater than 50 kDa were poorly preserved by immunoblotting compared to immunoprecipitation; however, below this threshold both techniques were equally efficient. Under the conditions of assay, it is probable that immunoprecipitation allows effective detection of both sequence and conformational determinants, whereas immunoblotting favours the detection of sequence determinants alone.
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In vitro exposure of adult rat alveolar macrophages to either paraquat or diquat resulted in concentration dependent cytotoxicity (cell death). The herbicide paraquat, however was statistically significantly more potent toward these cells than was diquat. The LC50 value for paraquat (8-h exposure, 37 degrees C) was determined to be 0.94 mM [95% confidence interval (C.I.) 0.79-1.12 mM], whereas the corresponding LC50 value for diquat was 1.97 mM (C.I. 1.58-2.51 mM). Interestingly, diquat was shown to enter these cells to a much greater extent than was paraquat. The latter data, while seemingly contradictory to the above findings, is consistent with other reported findings in this study that show that cell respiration, as measured by loss of oxygen consumption, was more sensitive to diquat than it was to paraquat. Also, only paraquat cytotoxicity was found to be dependent on oxygen tension and could be altered by the presence of antioxidant enzymes in the culture medium. Both compounds, however, were found to be equipotent toward purified mitochondria. Both paraquat and diquat were able to uncouple oxidative phosphorylation and induce active oxygen species (superoxide anions and hydrogen peroxide) from this organelle. It is concluded that free-radical pathology is the most likely mechanism of action by which paraquat is cytotoxic toward these cells, but that diquat poisoning probably originates from some other mode of action.
A 19-year-old woman presented with macroglossia of five months' duration and without bullae or erosions. A biopsy revealed a picture consistent with pemphigus vulgaris. The patient subsequently developed typical oral erosions. Her tongue enlargement and oral ulcerations improved dramatically with topical and systemic steroid therapy. Pemphigus vulgaris should be considered in the differential diagnosis of macroglossia.
When the herbicide paraquat (1,1'-dimethyl-4,4'-bipyridylium) was administered to adult rat pulmonary alveolar macrophages (PAM) in primary culture, both a time-dependent and a dose-dependent cytotoxic response (cell death) was observed. An LD50 value of 1 mM was calculated when these cells were exposed to paraquat in vitro for 12 h in Ham's F12 culture medium at 30 degrees C. Cell death was accompanied by the formation of TBA-reactive substances (lipid peroxidation) and was potentiated by hyperoxia (95% O2). In a 95% O2-5% CO2 atmosphere, an LD50 value of 0.1 mM was calculated. In addition, the presence of superoxide dismutase in the culture medium (1700 units/ml) inhibited the cytotoxic response. Since [14C]paraquat was not absorbed into these cells, extracellular superoxide anion radical formation was investigated as the cause of the observed cell death. Paraquat (0.5 mM) was found to stimulate extracellular O-2 generation, from PAM, but only in nonactivated cells. A sevenfold enhancement over the resting rate of radical generation was observed in the presence of paraquat. No increase in the O-2 generation rate of activated macrophages was observed upon the addition of paraquat to the culture medium. These data indicate that paraquat is cytotoxic to the pulmonary alveolar macrophage and further suggest that this cytotoxicity is mediated, at least in part, by an excess, extracellular production of active oxygen species. Implications of these findings with respect to the currently accepted hypothesis of paraquat poisoning in vivo are discussed.
We report a patient with a hyperpigmented, non-hairy plaque on the forearm. The presence of disorganized smooth-muscle bundles within the dermis proved this lesion to be a dermal smooth-muscle hamartoma. The relationship of this lesion to the pigmented hairy nevus of Becker is discussed.
Psoralen photoproducts were identified in association with cataracts in a psoriatic patient several years after oral methoxsalen plus ultraviolet A light (PUVA) therapy. Patients should be required to wear adequate optical protection not only during PUVA therapy but 24 hours following ingestion of methoxsalen.