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Biomedical subjects

R K Gupta

Publications and source records attributed to R K Gupta.

At least 19 recordsLinked to original sources

Lead inhibits 1,25-dihydroxyvitamin D-3 regulation of calcium metabolism in osteoblastic osteosarcoma cells (ROS 17/2.8).

We have determined the dose-response of 1,25-dihydroxyvitamin D-3 (1,25-(OH)2D3) on the intracellular free calcium-ion concentration ([Ca2+]i) in the osteoblastic osteosarcoma cells, ROS 17/2.8, using 19F-NMR and the intracellular divalent cation indicator, 1,2-bis(2-amino-5-fluorophenoxy)ethane-N,N,N',N'-tetraacetic acid (5F-BAPTA). The dose-response demonstrated an inverted U-shaped relationship with maximal elevation of [Ca2+]i at doses of 1 to 10 nM 1,25-(OH)2D3. At 10 nM, 1,25-(OH)2D3 elevated the [Ca2+]i from a control level of 118 +/- 4 nM to a peak value of 237 +/- 8 nM within 40 min. 1,25-(OH)2D3 also increased the initial rate of Ca2+ influx into ROS 17/2.8 cells, measured by 45Ca uptake, with a dose-response relationship which paralleled its effect on [Ca2+]i. Treatment of ROS 17/2.8 cells with Pb2+ at 1 and 5 microM significantly increased [Ca2+]i but significantly reduced the 1,25-(OH)2D3-induced elevation of [Ca2+]i. Simultaneous treatment of naive cells with 1,25-(OH)2D3 and Pb2+ produce little reduction of 1,25-(OH)2D3-induced 45Ca uptake while 40 min treatment with Pb2+ before addition of 1,25-(OH)2D3 significantly reduced the 1,25-(OH)2D3-induced increase in 45Ca influx. These findings suggest that Pb2+ acts by inhibiting 1,25-(OH)2D3-activation of Ca2+ channels and interferes with 1,25-(OH)2D3 regulation of Ca2+ metabolism in osteoblastic bone cells.

Animals

Cocaine induces intracellular free Mg deficits, ischemia and stroke as observed by in-vivo 31P-NMR of the brain.

31P-NMR spectroscopic studies were performed in vivo on brains of rats administered cocaine. Cocaine.HCl (1-5 mg/kg) administered systemically to lightly anesthetized rats resulted in significant and progressive deficits in whole brain intracellular free Mg ([Mg2+]i). Intracellular pH (pHi) also fell in a progressive manner but only after a significant fall in brain [Mg2+]i was noted. Both [Mg2+]i and pHi returned to normal in most rats. Brains of rats that exhibited stroke-like events, however, demonstrated continued intracellular acidosis associated with progressive loss of phosphocreatine and elevation of Pi up until death. These observations are consistent with the tenet that injection of cocaine can result in severe cerebral vasospasm, ischemia and rupture of cerebral blood vessels as a consequence of depletion of brain [Mg2+]i.

Animals

Needle aspiration cytology, immunocytochemistry, and electron microscopic study of unusual pancreatic carcinoma with pleomorphic giant cells.

Needle aspiration cytology, immunocytochemistry, and electron-microscopic findings are presented in three cases of an unusual pancreatic carcinoma in which pleomorphic giant cells formed an integral part of the tumour. All three patients were elderly males (age range 66-83 years) and had pancreatic masses. Notable cytologic features in all cases were the presence of bizarre mononucleated and multinucleated, poorly cohesive tumour giant cells, rare spindle cells, with occasional cannibalism and cytophagocytosis. Immunocytochemical study of aspirated material showed diffuse staining of cytokeratin and EMA within the tumour cells, while B 72.3 was seen as focal trace stain. Electron microscopy of aspirated material demonstrated epithelial features and these were characterised by the presence of tonofilaments and surface microvilli. Based on our findings, it is felt that the bizarre giant cells in this unusual variant of pancreatic carcinoma are of epithelial origin. The differential diagnosis of other tumours that may be associated with predominant giant cells in pancreatic aspirates is appropriately discussed.

Aged

Monoclonal antibody-based ELISA to detect glycoprotein tumor-associated-antigen-specific immune complexes in cancer patients.

This report describes the development and applicability of a tumor-associated-antigen-specific immune complex (IC) detection assay to denote the presence of tumor cells in a cancer host. This assay utilizes a murine monoclonal antibody, AD1-40F4, which was produced to a glycoprotein tumor-associated antigen (TAA). In Western blot, the murine monoclonal antibody recognized a 90-100 kD subunit of the antigen. This antigen is immunogenic in cancer patients and induces formation of endogenous antigen-antibody complexes. Analysis of 250 sera from normal individuals and 419 sera from cancer patients revealed that a significantly (P less than .0005) greater proportion (234/419; 55.9%) of cancer sera was positive for the marker than the normal sera (8/250; 3.2%). The incidence of the 90 kD-TAA-specific-IC was consistently and significantly higher in melanoma (58.5%; 38/65), sarcoma (52.9%; 83/157), and carcinoma of breast (50.9%; 58/114), lung (68.2%; 30/44), and colon (64.1%, 25/39) than in the normal group. The age of serum donors did not affect the incidence of the reactivity. Also, at least in the cancer group the gender of the serum donor did not affect the incidence of the 90 kD-TAA-specific-IC positivity. In a retrospective study where sequential serum samples obtained postoperatively from 105 patients with melanoma were analyzed, the 90 kD-TAA-specific-IC could be detected in 72 (69%) of patients several years before the appearance of clinically detectable disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent

A protease-sensitive site in the proposed Ca(2+)-binding region of human serum amyloid P component and other pentraxins.

Serum amyloid P component (SAP) is a decamer of 10 identical 25.5-kDa subunits. Limited proteolysis of SAP with alpha-chymotrypsin cleaves the subunit into two fragments of 18 and 7.5 kDa, although the fragments stay together in the decamer under nondenaturing conditions. Proteolysis does not occur in the presence of Ca2+ (10 mM). Cleavage with alpha-chymotrypsin prevents the Ca(2+)-dependent binding of SAP to zymosan extract, nucleosomes, and DNA. The alpha-chymotrypsin cleavage site identified is in a region of SAP that is highly conserved in members of the human C-reactive protein (CRP) family of proteins (pentraxins) to which SAP belongs and is similar to the Ca(2+)-binding site in calmodulin and related Ca(2+)-binding proteins (Nguyen, N.Y., Suzuki, A., Boykins, R.A., & Liu, T.-Y., 1986, J. Biol. Chem. 261, 10456-10465). Treatment of SAP with other proteases (trypsin, Pronase, and Nagarse protease) yields fragmentation patterns upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) that are similar to those obtained with alpha-chymotrypsin. Two other members of the pentraxin family of proteins, hamster female protein and rabbit CRP, also exhibit similar fragmentation patterns on SDS-PAGE when treated with the various proteases. Recently, it has been shown that the homologous protein, human CRP, is cleaved in the same homologous position as cleavage of SAP by alpha-chymotrypsin, resulting in the loss of Ca(2+)-binding (as shown by equilibrium dialysis) and Ca(2+)-dependent binding reactivities (Kinoshita, C.M., Ying, S.-C., Hugli, T.E., Siegel, J.N., Potempa, L.A., Jiang, H.J., Houghten, R.A., & Gewurz, H., 1989, Biochemistry 28, 9840-9848).(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence

Cine MR voiding cystourethrogram in adult normal males.

Cine voiding cystourethrogram was performed in 15 normal subjects using T1-weighted Turbo FLASH sequence. It showed the sequence of emptying of bladder, detrusor muscle contraction, opening of the sphincters, and various parts of urethra. It was found to be a good noninvasive test to study the dynamics of lower urinary tract without disturbing the state of the physiology.

Adult

p53 expression in Reed-Sternberg cells of Hodgkin's disease.

Mutation of the p53 protein may represent the commonest genetic event in human malignancy. Abnormal p53 expression has been reported in a variety of carcinomas, sarcomas and lymphoid neoplasms; however there is little information in relation to Hodgkin's disease. The expression of the nuclear phosphoprotein was investigated in paraffin-embedded biopsies from fifty patients with Hodgkin's disease using a polyclonal antibody, CM-1 and in snap-frozen material with monoclonal antibodies, PAb 1801 and PAb 240. Specifically, immunoreactivity was localised to the Reed-Sternberg cells or mononuclear variants in both nodular sclerosing (86% cases) and mixed cellularity (57% cases) subtypes of Hodgkin's disease. However, no positive staining was found in our cases of nodular lymphocyte predominant type Hodgkin's disease. Serial biopsies following recurrence of disease demonstrated consistent results. It is suggested that overexpression of p53, probably mutant, may have a role in the tumorigenesis of Hodgkin's disease.

Hodgkin Disease

Nutritional modulators of lipoprotein metabolism in patients with risk factors for coronary heart disease: diet and moderate exercise trial.

To study the role of diet as a risk factor for coronary heart disease (CHD), 463 adults (25-65 years, 419 males) at risk of or with CHD were assigned to group A (n = 231) receiving a cardiovasoprotective diet or group B (n = 232) receiving a normal diet in a randomized, single-blind fashion. Age, sex, risk factors and incidence of CHD were comparable between the two groups. Group A received a significantly higher percentage of calories in relation to complex carbohydrates, vegetable proteins, polyunsaturated fatty acids, and had a higher polyunsaturated:saturated fatty acid (P:S) ratio compared to control group B, which received more saturated fat and cholesterol. Group A also received more soluble dietary fiber and magnesium (Mg) and was physically more active than group B. Exercise and dietary adherence were tested by a questionnaire. After 12 weeks, results indicated a significant decrease in mean serum total cholesterol (9.0 vs 3.1%), low-density-lipoprotein cholesterol (LDL-C) (9.8 vs 3.8%) and triglyceride (11.1 vs 5.4%), and an increase in serum Mg (8.5%) in the intervention group compared to initial levels. No significant changes in mean levels were noted in the control group. Body weight significantly decreased and physical activity was significantly higher in the intervention group compared to the control group during the 12-week followup on similar intakes of energy. There were insignificant changes in high-density-lipoprotein cholesterol (HDL-C) in both groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

NMR measurement of intracellular potassium in the perfused normotensive and spontaneously hypertensive rat aorta by a multinuclear subtraction procedure.

Intracellular free potassium ion concentrations have been measured in the isolated perfused aortae from Wistar-Kyoto and spontaneously hypertensive rats by a novel multinuclear NMR procedure, which allows the estimation and subtraction of the extracellular potassium signal from the total 39K+ resonance. When tested on a sample of erythrocytes, the method yielded an intracellular potassium concentration of 149 mmol/L cell water, which is similar to the previously reported average of 140 mmol/L cell water, indicating full (100%) NMR "visibility" of red cell 39K+. In contrast, the 39K+ resonance of the perfused rat aorta preparation indicated very low (approximately 15%) NMR visibility of intracellular potassium in the aortic tissue, similar to that previously reported for perfused hearts. More importantly, aortae from spontaneously hypertensive rats yielded a corrected (for low NMR visibility) intracellular potassium concentration of only 90 +/- 10 (mean +/- SE; n = 6) mmol/L cell water, which was significantly lower (40%; P = .02) than that in aortae from normotensive rats. The NMR data thus reveal a significant depletion of intracellular potassium in hypertensive arterial smooth muscle cells.

Animals

Effect of the repellent deet on the antennal chemoreceptors for oviposition in Aedes aegypti (Diptera: Culicidae).

The insect repellent, N,N-diethyl-m-toluamide (deet), interferes with detection of the host attractant, lactic acid (LA), by LA-excited neurons on the antennae of female Aedes aegypti (L.) mosquitoes. In the current laboratory behavioral experiment, the attraction of gravid female mosquitoes to a source of ethyl propionate, an oviposition attractant, was reduced by a factor of two when deet was present. Similarly, when deet was presented together with ethyl propionate in electrophysiological experiments, sensitivity of the neural response to ethyl propionate was depressed in a transient, dose-dependent manner that could account for the reduction in behavioral attraction of gravid females to ethyl propionate. Electrophysiologically, the sensitivity of oviposition attractant receptors to ethyl propionate on gravid females was not different from that on nongravid host-seeking females. The similarity between the depression of sensitivity of the oviposition attractant-sensitive neurons induced by deet and that observed in the host attractant-sensitive neurons suggest a common mode of action of deet on peripheral chemoreceptor neurons that mediate these behaviors.

Aedes

Prolongation of survival in metastatic melanoma after active specific immunotherapy with a new polyvalent melanoma vaccine.

A new polyvalent melanoma cell vaccine (MCV) was administered to 136 stage IIIA and IV (American Joint Committee on Cancer) melanoma patients. Induction of cell-mediated and humoral immune responses to common melanoma-associated antigens present on autologous melanoma cells was observed in patients receiving the new MCV. This was accompanied by increased activation of tumor-infiltrating lymphocytes. Survival correlated significantly with delayed cutaneous hypersensitivity (p = 0.0066) and antibody responses to MCV (p = 0.0117). Of 40 patients with evaluable disease, nine (23%) had regressions (three complete). From our historical database of 126 stage IIIA and 1275 stage IV melanoma patients, there were no significant changes in the natural history of metastatic melanoma during the past 20 years. Univariate and multivariate analyses demonstrated prognostic significance for site of metastases (p = 0.0001) and immunotherapy with the new MCV (p = 0.0001). Overall our new MCV increased the median and 5-year survival of stage IIIA melanoma patients with regional soft tissue metastases twofold (p = 0.00024), and stage IV patients threefold (p = 0.0001) compared with previous immunotherapy and other treatments.

Adult

Fine needle aspiration of breast carcinoma in a fibroadenoma.

A case of concurrent carcinoma of the breast with a fibroadenoma in a 59-year-old woman is described. The diagnosis was made on fine needle aspiration cytology. Despite the rarity of such a lesion, it is felt that a cytological diagnosis of this lesion from a fine needle aspirate specimen is possible.

Adenofibroma

Coexpression of vimentin, cytokeratin and S-100 in monomorphic adenoma of salivary gland; value of marker studies in the differential diagnosis of salivary gland tumours.

An unusual coexpression of glial fibrillary acid protein (GFAP), keratin and vimentin occurs in pleomorphic adenoma of salivary gland. We designed this study to see if coexpression of the markers was also present in monomorphic adenoma of the salivary gland and whether monomorphic adenoma could be distinguished from other salivary gland tumours by marker studies. Immunocytochemical markers were used on fine needle aspiration samples from four cases of monomorphic adenoma, two cases of oncocytic adenoma, three cases of adenoid cystic carcinoma and four cases of pleomorphic adenoma. While positivity for cytokeratin, vimentin and S-100 was consistently found in all cases of monomorphic adenoma, only cytokeratin was expressed in adenoid cystic carcinoma. In pleomorphic adenoma, GFAP, cytokeratin and vimentin were coexpressed while in cases of oncocytic adenoma none of the markers was localized. Thus, it appears that by using a combination of GFAP, cytokeratin, vimentin and S-100 a distinction between these neoplasms may be possible. However, a larger study is needed to establish the usefulness of this approach.

Adenoma

Fine needle aspiration cytodiagnosis of apocrine carcinoma of the breast.

The cytopathological appearances of 14 cases of apocrine breast carcinoma diagnosed by fine needle aspiration cytology are described and the features compared to those seen in apocrine cells aspirated from benign cystic and solid lesions. Significant atypia must be observed before a diagnosis of apocrine carcinoma can be entertained.

Biopsy, Needle

Fine needle aspiration cytodiagnosis of a carcinoid tumour of the male breast.

The diagnosis of a primary carcinoid tumour of the breast in a 38-year-old male was initially made from a fine needle aspiration sample. The cytodiagnosis was supported by the histochemical demonstration of Grimelius-positive granules in tumour cells which were also found in tissue removed in a subsequent biopsy. An immunocytochemical study using markers for neuron-specific enolase and chromagranin in the aspirated sample and tissue was found to be negative. A total mastectomy with axillary node dissection was performed which showed no residual or metastatic tumour. No primary tumour was found elsewhere.

Adult

Na+ alters the affinity for glucose and phosphate in rat renal brush-border membranes: a study of NMR relaxation rates.

Transport of D-glucose (Glc) and Pi across renal microvilli brush-border membrane vesicles (BBMV) is coupled to Na+.Na+ may alter substrate binding and dissociation or the mobility of the translocator and its complexes. To evaluate substrate binding in NaCl or KCl solutions containing BBMV, we measured nuclear magnetic resonance transverse relaxation rates (1/T2) of [1-13C]Glc and 31P(i), as well as their temperature, magnetic field, and pH dependencies. The data indicate the following: 1) the alpha-anomer, but not the beta-anomer, of Glc binds to BBMV, more in the presence of Na+ than of K+; 2) interactions of P(i) with BBMV that increase its 1/T2 are significant in the presence of K+ but not of Na+; 3) temperature and magnetic field dependencies of the 1/T2 of P(i) bound to BBMV are consistent with more rapid exchange with free P(i) in the presence of Na+ than of K+; and 4) in the presence of Na+, [H+] promoted interactions of P(i) with BBMV that increase its 1/T2, probably by interfering with formation of rapidly dissociating Na(+)-P(i)-carrier complexes. In conclusion, Na+ promotes association of alpha-Glc to BBMV but promotes rapid exchange and dissociation of P(i) from BBMV. In both cases, Na+ effects coupling by altering the affinity of the cotransporters for the substrate.

Animals