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Biomedical subjects

S D Gupta

Publications and source records attributed to S D Gupta.

At least 19 recordsLinked to original sources

Morphometric image analysis and eosinophil counts in human liver allografts.

Histology of liver allografts is the gold standard for diagnosis of acute cellular rejection. However, scoring the severity of rejection and distinguishing it from other infiltrations is not easy. Only one group has evaluated biopsies morphometrically and also suggested that eosinophils are a specific diagnostic feature. We quantitated eosinophil count in 92 biopsies in a group of 25 patients and, in another group of 30 patients, used morphometric image analysis to measure the cross-sectional area and cell density in each portal tract in day 5 protocol liver biopsies. Rejection was diagnosed by pathological evaluation confirmed with clinical and biochemical graft dysfunction graded histologically into mild or moderate-to-severe. The control groups were five patients with no rejection, nine patients with CMV infection, and eight biopsies in eight patients for whom the cause of the liver dysfunction was obscure. The cross-sectional area, the inflammatory cell count of each portal tract and the mean portal tract inflammatory cell density (cells/mm2) increased with the severity of rejection. In each case the regression coefficient was statistically significant. Correlating the mean of the total inflammatory cell count with the mean of the portal inflammatory cell density (cell/mm2) gave far better separation of the mild rejection and moderate-to-severe rejection groups. Eosinophils were specific for the presence of acute cellular rejection and increased with the severity of rejection. They were absent in the no rejection group, in the CMV group and in those with obscure liver dysfunction. The eosinophil count fell markedly following treatment of rejection.(ABSTRACT TRUNCATED AT 250 WORDS)

Eosinophils

Primitive neuroectodermal kidney tumor: 2 case reports and review of the literature.

Peripheral neuroectodermal tumors are uncommon cancers arising from outside the central nervous system. The urinary system is rarely involved. The differentiation of these tumors from other small cell cancer and neuroblastoma is based on immunohistochemical differences. We report 2 cases of such tumors arising from the renal parenchyma. Tumor behavior and treatment modalities are discussed.

Adult

Identification of cutC and cutF (nlpE) genes involved in copper tolerance in Escherichia coli.

It has been suggested previously that copper transport in Escherichia coli is mediated by the products of at least six genes, cutA, cutB, cutC, cutD, cutE, and cutF. A mutation in one or more of these genes results in an increased copper sensitivity (D. Rouch, J. Camakaris, and B. T. O. Lee, p. 469-477, in D. H. Hamer and D. R. Winge, ed., Metal Ion Homeostasis: Molecular Biology and Chemistry, 1989). Copper-sensitive cutC and cutF mutants were transformed with a genomic library of E. coli, and copper-tolerant transformants were selected. Two distinct clones were identified, each of which partially restores copper tolerance in both the cutC and cutF mutants of E. coli. Subcloning, physical mapping, and sequence analysis have revealed that the cutC gene is located at 42.15 min on the E. coli genome and encodes a cytoplasmic protein of 146 amino acids and that the cutF gene is located at 4.77 min on the E. coli genome and is allelic to the nlpE gene independently identified by Silhavy and coworkers (W. B. Snyder, L. J. B. Davis, P. N. Danese, C. L. Cosma, and T. J. Silhavy, J. Bacteriol. 177:4216-4223, 1995). Results from the genetic mapping of the copper-sensitive mutations in the cutF mutant and sequencing of the cutC and cutF (nlpE) alleles from both cutC and cutF mutants indicate that both the cutC and cutF mutants are in fact double mutants altered in these two genes, and mutations in both the genes appear to be required for the copper-sensitive phenotype in each mutant.

Alleles

Phosphatidylethanolamine is not essential for the N-acylation of apolipoprotein in Escherichia coli.

It has been postulated that the N-acyl fatty acid attached to the amino terminus of the major Escherichia coli lipoprotein is derived from the fatty acid at the 1-position of phosphatidylethanolamine (PtdEtn) (Jackowski, S., and Rock, C.O. (1986) J. Biol. Chem. 261, 11328-11333). To ascertain the role of PtdEtn in the conversion of apolipoprotein to the mature lipoprotein, the lipoprotein from E. coli strain AH930 (pss::kan) containing a null mutation in the phosphatidylserine synthase gene (pss) was studied. Pulse labeling with [35S]methionine for 30 s or 5 min revealed the formation of mature lipoprotein in both wild-type (W3110) and mutant (AH930) cells. [3H]Palmitate-labeled lipoproteins from both the mutant and wild-type cells were found to contain nearly identical amounts of alkali-resistant (amide-linked, 41-42%) and alkali-labile (ester-linked, 58-59%) fatty acids. Edman degradation and dansylation of the immuno-affinity-purified [35S]cysteine-labeled lipoprotein showed that the NH2 terminus of the lipoprotein in the mutant was blocked as in the wild type. In vitro assay of apolipoprotein N-acyltransferase using membranes either from the mutant or the wild-type strain as the source of both the enzyme and the acyl donor revealed that both membranes were equally active in the conversion of [35S]methionine-labeled apolipoprotein to lipoprotein. These data strongly suggest that PtdEtn is not essential for the N-acylation of apolipoprotein to form lipoprotein, and other major phospholipids such as phosphatidylglycerol and cardiolipin can serve as the donor of fatty acid in the N-acylation of apolipoprotein.

Acylation

Identification and subcellular localization of apolipoprotein N-acyltransferase in Escherichia coli.

Apolipoprotein N-acyltransferase, the enzyme catalyzing the conversion of apolipoprotein to mature lipoprotein, was detected by an in vitro assay using [35S]methionine-labeled apolipoprotein as the substrate. Triton X-100 solubilized the enzyme, and was required for its activity. The enzyme showed a broad pH optimum (pH 6.5-7.5). N-Acylation of apolipoprotein with ethanol-washed membranes was dependent on exogenous phospholipids, with phosphatidylethanolamine, phosphatidylglycerol and cardiolipin all showing about 10- to 20-times enhancement of the enzyme activity in the delipidated membranes. Incubation of apolipoprotein with [3H]palmitate-labeled membranes resulted in the incorporation of [3H]palmitate into lipoprotein. The enzyme was found to be enriched in the inner membrane and in the inner membrane/outer membrane mixed fractions of the E. coli cell envelope.

Acyltransferases

Histochemical pattern in alimentary tract duplications of children.

Detailed histological and histochemical studies using PAS, AB-PAS, and high-iron diamine-AB were done on 12 alimentary tract duplication cysts in children. Nine of these were thoracic and three were abdominal in location. A total of 11 cysts (nine thoracic and two abdominal cysts) were lined by gastric mucosa seen in varying stages of maturation. One triloculated cyst had, in addition, small intestinal mucosa and bronchial wall mucosa. One intra-mucosal rectal cyst was lined exclusively with primitive rectal mucosa. The cysts showed a variable pattern of mucin histochemistry. In three cysts, the gastric mucosa showed neutral mucins. In another three cysts, in addition to neutral mucins, focal positivity for sulphomucins was observed in the apical region of the surface epithelial cells. In the remaining five cysts, the gastric mucosa showed neutral mucins along with a variable mixture of sulpho- and sialomucins. A correlation was seen between mucin histochemistry and the age of the patient. Neutral mucins or focal positivity for sulphomucins was seen in patients aged 7 months or more, whereas a mixture of sulpho-, sialo-, and neutral mucins was identified in patients who were 1 month old, or less.

Abdomen

Adult embryonal rhabdomyosarcoma of the vagina complicating pregnancy: a case report and review of the literature.

Embryonal rhabdomyosarcoma of the vagina in adults per se is uncommon and pregnancy complicated by such tumors is extremely rare. We wish to report one case of vaginal embryonal rhabdomyosarcoma, diagnosed at the age of 23 years who was carrying at the time of initial presentation. The problem of management of rhabdomyosarcoma in adults, particularly when they are associated with pregnancy is discussed. We advocate the use of multiagent chemotherapy and pelvic radiotherapy in its overall management.

Adult

The biosynthesis of sulfoquinovosyldiacylglycerol: studies with groundnut (Arachis hypogaea) leaves.

The biosynthetic pathway of sulfoquinovosyldiacylglycerol (SQDG) was investigated using groundnut (Arachis hypogaea) leaf discs and 35S-labeled precursors. [35S]SO4(2-) was actively taken up by the leaf discs and rapidly incorporated into SQDG. After 2 h, 1.5% of the [35S]SO4(2-) added to the incubation medium was taken up, of which 28% was incorporated into SQDG. The methanol-water phases of the lipid extracts of the leaf discs were analyzed for the 35S-labeled intermediates. Up to 2 h of incubation, cysteic acid, 3-sulfopyruvate, 3-sulfolactate, 3-sulfolactaldehyde, and sulfoquinovose (SQ) which have been proposed as intermediates [Davies et al. (1966) Biochem. J. 98, 369-373] were not labeled. Only a negligible amount of radioactivity was observed in these compounds after incubation for 4 h and more. Addition of sodium molybdate inhibited the uptake of [35S]SO4(2-) as well as its incorporation into SQDG by the leaf discs, suggesting that 3'-phosphoadenosine-5'-phosphosulfate may be involved in the biosynthesis of SQDG. Addition of unlabeled cysteic acid to the incubation medium enhanced the uptake of [35S]SO4(2-) but did not affect its incorporation into SQDG. 35S-labeled cysteic acid was taken up by the leaf discs and metabolized to sulfoacetic acid but not incorporated into SQ or SQDG. These results show that cysteic acid is not an intermediate in SQDG biosynthesis. [35S]SQ was taken up by the leaf discs and incorporated into SQDG in a time-dependent manner. [35S]Sulfoquinovosylglycerol was also taken up by the leaf discs but not incorporated into SQDG. It is concluded that SQDG is not biosynthesized by the proposed sulfoglycolytic pathway in higher plants. Though [35S]SQ was converted to SQDG, the rates are much lower compared to [35S]SO4(2-) incorporation, which suggests that a more direct pathway involving sulfonation of a lipid precursor may exist in higher plants.

Arachis

Metabolism of the plant sulfolipid--sulfoquinovosyldiacylglycerol: degradation in animal tissues.

Metabolism of the plant sulfolipid--sulfoquinovosyldiacylglycerol (SQDG)--was studied in animal tissues. In vivo experiments with [35S]SQDG in guinea pigs showed that this lipid is not absorbed intact in the gastrointestinal tract. In these experiments, 3 h after administration of [35S]SQDG, the intestinal mucosa contained 1 to 5% of the radioactivity as SQDG, while the remainder was in a water-soluble form. Analysis of the water-soluble components showed that about 60% of the radioactivity was present as sulfoquinovosylglycerol (SQG) and the remainder was present as free SO4(2-). In the blood, 99% of the radioactivity was present as SO4(2-), SQG was not observed. In liver, only very little radioactivity was observed and appeared to be mainly in the form of SO4(2-). Experiments with everted intestinal sacs of guinea pigs confirmed the formation of SQG, SO4(2-), and, in addition, sulfoquinovosylmonoacylglycerol (SQMG) in this tissue. In vitro experiments with saline extracts of acetone powders of pancreas and intestinal mucosa of guinea pig, sheep, and rat showed that [35S]SQDG was deacylated to SQMG (sulfolipase A activity) and SQG (sulfolipase B activity). It is concluded that animal tissues deacylate SQDG in a stepwise manner to SQG. It is further metabolized to yield free SO4(2-) by cleavage of the C-S bond which appears to be brought about by the intestinal microflora. Sheep pancreatic sulfolipases were characterized. Bile salts, sodium dodecyl sulfate, and Triton X-100 inhibited the pancreatic sulfolipases, while CaCl2 activated them. Substrate competition experiments and investigations on substrate specificity with a partially purified preparation indicated that relatively specific sulfolipase(s) may exist in pancreas. Among the species tested, guinea pig tissues showed the highest sulfolipase A and B activities followed sheep and rat tissues. Pancreatic enzymes were 18 to 60 times more active than intestinal enzymes.

Animals

Endoscopic evaluation of dysphagia in the elderly.

A survey of 100 consecutive endoscopies on elderly patients with suspected obstructive dysphagia is reported. Seventy-eight patients had positive findings and one to three lesions were reported in these cases. Benign stricture of the oesophagus was the commonest finding, and led to active treatment (dilation) in most cases either with Hurst bougies or Eder dilators. Fifteen patients had upper gastro-intestinal malignancy (12 oesophagus, three stomach). All of those with oesophageal carcinoma received some form of active treatment, either in the form of radiotherapy, surgery, dilation or Celestin tube insertion. Six patients with negative endoscopies had diagnoses established by other means and in 14 no cause was identified, but in four of these symptoms settled after endoscopy. The duration of dysphagia was not particularly helpful clinically. Two thirds of 'endoscopically normal' patients had short histories with dysphagia for less than 1 year, as did half of the patients with benign stricture. Most patients who were found to be malignant had experienced dysphagia for only a short time although one patient had a 4-year history! Upper gastro-intestinal endoscopy is a safe and valuable procedure in elderly patients with dysphagia and often leads to positive therapeutic intervention even in quite frail subjects.

Aged

Changes of lipid spectrum in different tissues of Furadan-treated mice.

The effects of multiple intraperitoneal doses of Furadan (0.125 mg/kg, 0.25 mg/kg and 0.50 mg/kg, respectively) for the 2nd week, 4th week and 6th week of treatment on the concentrations of total lipid and its different fractions and lipase activity in mice were studied. The following were measured in liver, kidney, brain and serum; total lipid, cholesterol (total and free), phospholipid (total and its fractions--lecithin, lysolecithin, phosphatidyl ethanolamine and lysophosphatidylethanolamine), triglyceride, free fatty acid. Lipase activity was measured in liver and serum. Furadan caused biochemical disorders of the tissues. All the aforesaid lipid fractions (except free fatty acid) are elevated significantly in liver, kidney and serum of Furadan-treated mice. Free fatty acid increased significantly in serum and remain constant in other tissues of pesticide-treated mice. All the lipid fractions except phospholipid and its fractions remained constant in brain of treated mice; total phospholipid and its fractions decreased significantly in brain of treated mice. Lipase activity decreased significantly in liver and serum of treated mice. Such disorders of lipid levels in the aforesaid tissues might be associated with CNS depressant action and structural and functional toxicity of other tissues induced by Furadan.

Animals