The regulation of T-cell antigen receptor signal transduction function.
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Biomedical subjects
Publications and source records attributed to D Desai.
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We studied the adherence of Acanthamoeba to three types of new, unworn soft contact lenses: daily wear; extended wear; and disposable contact lenses. We used a human corneal isolate of Acanthamoeba castellanii in commercially prepared saline. Unworn lenses were exposed to Acanthamoeba cysts or trophozoites with stirring. After exposure the numbers of Acanthamoeba adhering were determined by light microscopy and confirmed by scanning electron microscopy. In general, the adherence of Acanthamoeba cysts and trophozoites to disposable lenses was less than that to daily wear and extended wear lenses. However, any adherence of Acanthamoeba to soft contact lenses is a potential source of corneal infection.
The phosphorylation of the invariant chains associated with the human TCR has been investigated after the stimulation of T lymphocytes with CD2 mAb T11(2) and T11(3), PHA, or phorbol 12,13-dibutyrate. As described previously, stimulation of T cells with either CD2 mAb or phorbol 12,13-dibutyrate resulted in the phosphorylation of the CD3 gamma-chain. The combination of T11(2) and T11(3) mAb also induced phosphorylation of the TCR zeta-chain. The phosphorylated zeta-polypeptide of CD2-activated cells was immunoprecipitated with antiphosphotyrosine antibodies and migrated to a 21- to 23-kDa position during SDS/PAGE. These results indicate that stimulation of human T cells via the CD2 Ag with the T11(2) and T11(3) mAb activates not only protein kinase C but also tyrosine kinase(s), resulting in the phosphorylation of the CD3 gamma-chain and the tyrosine phosphorylation of the zeta-chain, respectively.
Advancing age tends to be accompanied by predictable changes in organ-system function and body composition, as well as an increased prevalence of various diseases. Our knowledge of the metabolic responses to surgical stress is derived largely from studies of young and middle-aged individuals. Whether these responses are altered in the elderly is not well established; the characteristic changes in metabolic function and body composition that occur in the elderly may limit their ability to respond adequately and to survive severe or complicated surgical procedures. To evaluate the effects of age and differences in body composition on the metabolic responses to surgery, we studied 20 active and otherwise healthy men aged 43 to 77 years, before and after elective colon resection. Age was not related to weight or total body water (TBW) in the patients studied. Resting preoperative energy expenditure was strongly dependent on TBW (reflecting lean body mass) and age (r2 = 0.80, p less than 0.001), and to TBW and creatinine excretion (reflecting muscle mass) (r2 = 0.85, p less than 0.001). Energy expenditure increased 18 +/- 2% (range, 4% to 40%) after operation but this response was unrelated to age. Postoperative urine nitrogen was related to body weight or TBW, but not to age. Serum glucose, cortisol, white blood cell count count, and C-reactive protein responses were also independent of patient age. The metabolic responses of generally healthy men to a moderate surgical stress do not vary with age in the range studied. The metabolic responses to more severe or prolonged stress may be altered with advancing age and changes in body composition, and in patients with concomitant diseases.
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Contact lens-related Acanthamoeba keratitis has been associated with the use of soft contact lenses and homemade saline solutions. We studied the adherence of Acanthamoeba to unworn extended-wear soft contact lenses on a human corneal isolate of Acanthamoeba castellanii suspended in normal saline (cysts, 6.3 x 10(5)/ml; trophozoites, 3.6 x 10(5)/ml). Unworn hydrogel contact lenses (polymacon, water content 38.6%; 50 lenses, 400 lens segments) were exposed to A. castellanii cysts or trophozoites with or without stirring. After exposure for zero to seven hours, soft contact lenses were or were not washed, and the A. castellanii adherence was determined with a standardized light-microscopy technique (40 fields, ten lens segments for each count) and with scanning electron microscopy. Both cysts and trophozoites adhered to the unworn soft contact lenses. Trophozoites showed acanthopodia, lobopodia, and filopodia. This study indicates that both A. castellanii cysts and trophozoites can firmly adhere to unworn soft contact lens. Such adherence may play a role in the pathogenesis of soft contact lens-related Acanthamoeba keratitis.
The 3'-monophosphates of the major deoxyribonucleoside adducts of five representative polynuclear aromatic hydrocarbons (PAH)-benzo[a]pyrene, benz[a]anthracene, chrysene, 5-methylchrysene and 6-methylchrysene--were prepared by reaction with DNA of the appropriate bay-region diol epoxides followed by enzymatic hydrolysis and HPLC purification. These standards were converted to 3',[32P]-5'-bisphosphates as in the Randerath 32P-postlabeling assay. TLC and HPLC systems for their separation are described. HPLC conditions for separation of the corresponding 3'- and 5'-monophosphates are also reported. These data will be useful for determining whether unknown DNA adducts detected by the 32P-postlabeling assay may have resulted from reactions of DNA with PAH diol epoxides.
The metabolic responses to trauma and surgery have been well described and are observed most typically in otherwise healthy young and middle-aged individuals. To investigate the effect of age on blood glucose, insulin, and cortisol responses after mild to moderate trauma, we studied 33 patients (Injury Severity Scores, 5-38, and ages 16 to 81 years) before resuscitation and 24-96 hours postinjury. Age was associated with an increase in serum glucose during both "ebb" and "flow" phases of the injury response, but not with serum insulin. Serum glucose also increased with the degree of injury as reflected in Glasgow Coma Scale on admission and Injury Severity Score subsequently. Serum cortisol responses were increased in older patients and tended to decrease with time following injury. A more detailed knowledge of how age may alter the ability of elderly patients to respond to trauma and critical illness is essential to allow the continued development of rational therapies for such patients.
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Systemic protothecosis was diagnosed in a 7-year-old dog that had only ocular manifestations. During the 3-month course of disease, a variety of drugs was administered, including amphotericin B, gentamicin, and ketoconazole. The ocular signs initially abated, but subsequently worsened during this period. The dog was found dead 3 months after initial examination, and systemic protothecosis was confirmed at necropsy.
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trans-3'-Hydroxycotinine is a major urinary metabolite of nicotine in smokers, but no straightforward method is available for its synthesis. A simple method was developed for preparation of trans-3'-hydroxycotinine from cotinine in two steps by using NaN[(CH3)3Si]2 and dibenzyl peroxydicarbonate, followed by base-catalyzed hydrolysis.
In previous studies, we have observed unexpected structure-tumorigenicity relationships among the dimethylchrysenes. Thus, 5,6-dimethylchrysene and 5,7-dimethylchrysene were only weakly tumorigenic and were significantly less active than 5-methylchrysene. These results were surprising in view of the known route of metabolic activation of 5-methylchrysene via its 1,2-diol 3,4-epoxide. In this paper, we extended our studies of structure-tumorigenicity relationships among the dimethylchrysenes. We synthesized 5,7-, 5,8-, 5,9-, and 5,10-dimethylchrysene via photochemical ring closure reactions. The tumor-initiating activities of these dimethylchrysenes on mouse skin were compared with those of 5-methylchrysene and 5,6-dimethylchrysene. 5-Methylchrysene and 5,9-dimethylchrysene were highly tumorigenic and were significantly more active than 5,6-, 5,7-, 5,8-, and 5,10-dimethylchrysene. The results of these studies, taken together with those reported in the subsequent two papers, suggest that the molecular shapes of dimethylchrysenes influence the balance between metabolic activation and detoxification pathways.
A previously unknown urinary metabolite of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) was identified as 4-(methylnitrosamino)-1-[3-(6-hydroxypyridyl)]-1-butanone (6-hydroxyNNK). The metabolite was initially isolated from rat urine. On the basis of its MS and NMR, it was either a 4- or 6-hydroxypyridyl derivative of NNK. Model compounds were synthesized to distinguish between these possibilities; the results indicated that the metabolite was 6-hydroxyNNK. This was confirmed by independent synthesis; the spectral and chromatographic properties of 6-hydroxyNNK were the same as those of the metabolite. F-344 rats and A/J mice treated with 100 mg/kg NNK excreted approximately 1% of urinary metabolites as 6-hydroxyNNK; it was not detected as a sulfate or glucuronide conjugate. This is the first example of a pyridyl-hydroxylated metabolite of a tobacco-specific nitrosamine. On the basis of comparison to published data on other pyridine derivatives, 6-hydroxyNNK may be formed by bacterial metabolism. The potential utility of 6-hydroxyNNK as a dosimeter of human uptake of NNK is discussed.
The regioselective synthesis of 3-methoxybenz[a]anthracene-7,12-dione by oxidative photocyclization is described. The synthesis involved preparation of 1-(1-bromo-2-naphthyl)-2-(3- methoxyphenyl)ethylene by Wittig reaction, followed by photocyclization for 4 h. This gave 7-bromo-3-methoxybenz[a]anthracene. Extended photocyclization over 16 h under similar conditions gave 3-methoxybenz[a]anthracene-7,12-dione, an important intermediate in the synthesis of 7,12-dimethylbenz[a]anthracene-3,4-diol 1,2-epoxide.