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Biomedical subjects

R K Gupta

Publications and source records attributed to R K Gupta.

At least 613 records · Page 34Linked to original sources
PubMed1985

23Na NMR studies of rat outer medullary kidney tubules.

Two reservations have previously made interpretation of biological 23Na NMR measurements difficult: the "size" of the extracellular space penetrated by the shift reagent and the possibility of a 60% reduction in the intensity of the NMR-visible 23Na signal due to quadrupolar interactions (Berendsen, H. J. C., and Edzes, H. T. (1973) Ann. N. Y. Acad. Sci. 204, 459-485; Civan, M. M., Degani, H., Margalit, Y., and Shporer, M. (1983) Am. J. Physiol. 245, C213-C219; Gupta, R. K., and Gupta, P. (1982) J. Magn. Reson. 47, 344-350). We have addressed both these issues using a suspension of rat outer medullary kidney tubules, nephron segments responsible for the fine control of total body volume and electrolyte balance. First, the extracellular space penetrated by the shift reagent dysprosium tripolyphosphate, as defined by the extracellular 23Na resonance, revealed a space similar to that which contained extracellular 35Cl- ions. Measurement of an extracellular 35Cl- space using 35Cl NMR was possible because the intracellular 35Cl- resonance was broadened beyond detection in the cells studied. Second, to characterize the reduction of the 23Na signal by quadrupolar interactions, the intracellular 23Na level was raised artificially by simultaneously inhibiting Na+ efflux and increasing the ion permeability of the plasma membrane. Under these conditions, NMR-observable intracellular Na+ reached a level which was approximately 81% of that in the medium, a level determined using chemical techniques. This observation would suggest that the resonance of the intracellular 23Na pool was not subject to a 60% reduction in signal intensity, as a result of nuclear quadrupolar interaction. The intracellular 23Na level measured, under basal conditions, was 23 +/- 2 mumol/ml of cell water (37 degrees C) (n = 3, S.D.) and was demonstrated to be responsive to a number of physiological stimuli. The level was temperature-sensitive. It was reduced by inhibitors of apical Na+ transport, furosemide and amiloride, and it was raised with (Na+ + K+)-ATPase inhibition. The furosemide and amiloride actions described would suggest that the Na+-transporting mechanisms sensitive to these agents (e.g. Na+/K+/Cl- cotransport system, Na+:H+ exchange system) contribute to the regulation of the intracellular Na+ level in the kidney tubular preparation studied.

Adrenalectomy↗
PubMed1985

Studies of insulin action on the amphibian oocyte plasma membrane using NMR, electrophysiological and ion flux techniques.

Insulin (0.1-10 microM) reinitiates the meiotic divisions in Rana oocytes and produces a 14-20 mV negative-going hyperpolarization of the plasma membrane as well as a 0.25 unit increase in intracellular pH during the first 90 min. During hyperpolarization, the Na+ conductance of the membrane decreases by 40-50% with a concomitant increase in 22Na+ uptake from the medium. The increased uptake of Na+ during a period of decreasing Na+ conductance is apparently due to an increase in fluid phase turnover associated with insulin-mediated endocytosis. Both membrane hyperpolarization and increase in pHi are Na+-dependent and are blocked by the serine proteinase inhibitor, phenylmethylsulfonyl fluoride. The membrane potential of the prophase oocyte has a significant electrogenic component with potential but not conductance sensitive to glycosides and substitution of Li+ for Na+. Insulin hyperpolarizes Li+ or glycoside-treated oocytes whereas glycosides do not affect insulin-hyperpolarized oocytes. [3H]Ouabain binding by the plasma membrane of the untreated oocyte shows at least two K+-sensitive components (Kd = 42 and 2000 nM) linked to inhibition of the Na+ pump. Insulin-treated oocytes show a single class of intermediate-affinity ouabain sites (Kd = 490 nM) which appear to result from insulin-induced internalization of membrane-bound ouabain. [125I]Insulin binding to the plasma membrane shows a class of high-affinity sites (Kd = 87 nM) with 40-50 pump sites per insulin-binding site. Our results suggest that insulin-induced mediator peptides stimulate Na+-H+ exchange resulting in an increase in intracellular pH and Na+ uptake concomitant with an increase in receptor-mediated endocytosis and a decrease in Na+ conductance and associated membrane hyperpolarization. The net result appears to be a down-regulation of the Na+ pump which together with a decrease in Na+ conductance may divert high-energy phosphate compounds from cation regulation to anabolic processes of meiosis.

Animals↗
PubMed1985

NMR studies of intracellular sodium ions in mammalian cardiac myocytes.

The unambiguous measurement of intracellular sodium ion [Na+]i by the noninvasive NMR technique offers a new opportunity to monitor precisely the maintenance and fluctuations of [Na+]i levels in intact cells and tissues. The anionic frequency shift reagent, dysprosium (III) tripolyphosphate, which does not permeate intact cells, when added to suspensions of intact adult rat cardiac myocytes, alters the NMR frequency of extracellular sodium ions, [Na+]o, leaving that of intracellular ions, [Na+]i, unaffected. Using 23Na NMR in conjunction with this shift reagent, we have determined NMR-visible intracellular Na+ ion concentration in a suspension of isolated cardiac myocytes under standard conditions with insulin and Ca2+ in the extracellular medium to be 8.8 +/- 1.2 mmol/liter of cells (n = 4). This value is comparable to that measured by intracellular ion-selective microelectrodes in heart tissue. Cardiac myocytes incubated for several hours in insulin-deficient, Ca2+-containing medium prior to NMR measurement exhibited a somewhat lower [Na+]i value of 6.9 +/- 0.5 mmol/liter of cells (n = 3). Reversible Na+ loading of the cells by manipulation of extracellular calcium levels is readily measured by the NMR technique. Incubation of myocytes in a Ca2+-free, insulin-containing medium causes a 3-fold increase in [Na+]i to a level of 22.8 +/- 2.6 mmol/liter of cells (n = 10). In contrast to cells with insulin, insulin-deficient myocytes exhibit a markedly lower level of [Na+]i of only 14.6 +/- 2.0 mmol/liter of cells (n = 4) in Ca2+-free medium. These observations suggest that insulin may stimulate a pathway for Na+ influx in heart cells.

Animals↗
PubMed1985

The titration of tetanus antitoxin IV. Studies on the sensitivity and reproducibility of the toxin neutralization test.

The quality of tetanus toxin affected the sensitivity of the toxin neutralization (TN) test greatly. By using purified toxin a minimum level of 0.001 IU/ml of tetanus antitoxin could be detected whereas with crude toxin a level of 0.025 IU/ml only could be detected. The TN test described in this report permitted titration of tetanus antitoxin in twofold dilution steps from levels as low as 0.001 IU/ml using 0.6 ml of serum only at the L+/5000 level of purified tetanus toxin. Treatment of the sera with 2-mercaptoethanol (2-ME) did not affect the TN titres showing that the TN test detects the neutralizing antibodies (IgG) which are not affected by 2-ME. The TN test was found to be a highly sensitive and reproducible test.

Animals↗
PubMed1985

The titration of tetanus antitoxin V. Effect of formalization method for the fixation of sheep erythrocytes on the indirect haemagglutination test.

Two methods of fixation of sheep erythrocytes with formaldehyde for the titration of tetanus antitoxin by the indirect haemagglutination (IHA) test have been compared. The cells fixed with 3% formaldehyde at 4-8 degrees C for 24 h (formaldehyde (I) fixed cells) were less sensitive than the cells fixed with 3% formaldehyde at 4-8 degrees C for 24 h and subsequently treated with 40% formaldehyde at 4-8 degrees C for a further 24 h (formaldehyde (II) fixed cells). The correlation between the toxin neutralization (TN) and IHA titres using formaldehyde (I) fixed cells was better than that obtained with formaldehyde (II) fixed cells. There was no statistically significant difference between TN and IHA titres after treatment of the sera with 2-Mercaptoethanol using formaldehyde (I) fixed cells. Formaldehyde (I) fixed cells can be used for two months with adequate sensitivity to detect the minimum protective level of tetanus antitoxin in the sera.

Animals↗
PubMed1985

Experimental zinc deficiency in guinea-pigs: clinical signs and some haematological studies.

Thirty-three male, weanling guinea-pigs were divided into two groups. One group was fed on a zinc-deficient (ZnD) diet (1.25 mg Zn/kg diet) for 45 d and then divided into two subgroups: subgroup 1 continued on diet ZnD while subgroup 2 was fed on a Zn-repleted diet (100 mg Zn/kg diet) for 15 d. The second group was fed on a Zn-adequate diet (50 mg Zn/kg diet) for 60 d. Zn deficiency could be produced within 21 d as evidenced by clinical signs and reduction in serum Zn concentration. Clinical signs exhibited were listlessness, scabby lesions on skin near the foot pads, mild alopecia and a reduction in body-weight gain. No effect was observed on food intake. Significant decreases in packed cell volume and total leucocyte count from 48 d onwards accompanied by absolute lymphocytopenia and relative neutrophilia were observed. Supplementation of Zn in the Zn-repleted group resulted in marked, through incomplete, improvement with regard to serum Zn concentration, clinical signs and haematological changes.

Animals↗
PubMed1985

Characterization of antigenic components of immune complexes isolated from a melanoma patient.

Immune complexes (IC) from the serum of a melanoma patient were partially purified with DEAE Affigel Blue. The IC were radioiodinated and then immobilized with activated Sepharose 4B. Immunologically bound components in the IC were dissociated with 3M MgCl2 followed by gel filtration chromatography. A distinct antigen component of about 700,000 daltons was obtained by refractionation of a region of the chromatogram containing antigenic activity by Sephacryl S-300 column chromatography. Though the antigenic component behaved as a single entity in the gel filtration, it consisted of at least five polypeptide chains that were revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. However, 92,000 and 41,000 dalton subunits were predominant. This procedure may be useful for isolating tumor antigens from circulating IC of cancer patients.

Antigen-Antibody Complex↗
PubMed1985

Changes in intracellular Mg adenosine triphosphate and ionized Mg2+ during blood storage: detection by 31P nuclear magnetic resonance spectroscopy.

31P nuclear magnetic resonance (NMR) spectroscopy was used to measure changes in intra-erythrocyte Mg adenosine triphosphate (MgATP) and free Mg2+ during blood storage at 4 degrees C in standard citrate preservation media. The extent of Mg2+ complexation of ATP and the concentration of free Mg2+ were measured from the Mg2+-dependent chemical shift differences, at 22 degrees C, between the P beta and P alpha resonances of intracellular ATP. This difference changed from 721.0 +/- 1.4 Hz (mean +/- SE) on the day of collection to 741.0 +/- 3.4 Hz after three to seven days and 774.0 +/- 2.8 Hz after 11 to 40 days storage in either acid-citrate-dextrose (ACD) or citrate-phosphate-dextrose-adenine (CPDA-1). Changes in intracellular pH, detected from shifts in the intracellular Pi resonance, averaged 0.27 units after 11 to 40 days of storage. These data indicate a sizable decrease in the extent of Mg2+ complexation of ATP, and a decrease by a factor of 2.6 in free Mg2+, during the shelf-life of blood stored in ACD or CPDA-1.

Adenosine Triphosphate↗
PubMed1984

Role of calcium in regulating intracellular pH following the stepwise release of the metabolic blocks at first-meiotic prophase and second-meiotic metaphase in amphibian oocytes.

31P-NMR has been used to monitor changes in intracellular pH following the sequential release of the block at first-meiotic prophase by hormones and the block at second-meiotic metaphase by fertilization in Rana eggs and oocytes. The broad phosphoprotein signal was eliminated by a combination of spin-echo and deconvolution techniques. pHi was determined from the pH-dependent separation of intracellular Pi and phosphocreatine resonances. Agents that release the prophase block (progesterone, insulin, D-600, La3+) increased pHi from 7.38 to 7.7-7.8 within 1-3 h. Noninducers such as 17 beta-estradiol were without effect. By second-metaphase arrest (ovulated, unfertilized) the pHi had fallen to 7.1-7.2. pHi underwent a transient increase to about 7.7 within the first 30 min at fertilization, with a slow 0.1-0.2 pH unit oscillation during early cleavage. The progesterone-induced elevation of intracellular pH is not blocked by amiloride and occurs in Na+-free medium. A transient rise in pHi occurs when the prophase-arrested oocyte is transferred to Ca2+-free medium or when ionophore A23187 is added to the Ca2+-containing medium. Agents that inhibit the resumption of the first meiotic division either block the rise in pHi (procaine, PMSF) or shorten the time-course of the rise in pHi (ionophore A23187). Conditions that elevate intracellular Ca2+ levels and/or increase Ca2+ exchange produce an increase in pHi, whereas those conditions that decrease intracellular Ca2+ levels and/or exchange produce a fall in pHi within 1 h. The time-course of the increase in pHi both following release of the prophase block and at fertilization coincide with a fall in intracellular cAMP and release of surface and/or intracellular Ca2+. These results suggest that: (1) pHi is a function of cytosolic free Ca2+ levels and/or Ca2+ exchange across the oocyte plasma membrane, and (2) meiotic agonists (progesterone, insulin, D-600) and mitogens (sperm, ionophore A23187) modulate intracellular and/or membrane Ca2+ with the resulting changes in pHi and cAMP and resumption of the meiotic divisions.

Animals↗
PubMed1984

A prospective postoperative evaluation of urinary tumor-associated antigens in sarcoma patients. Correlation with disease recurrence.

Tumor-associated antigens (TAA) excreted into the urine of sarcoma patients can be detected by the complement-fixation assay. The authors prospectively measured TAA levels in the urine of 50 sarcoma patients postoperatively to determine whether reappearance of antigen in the urine would be predictive of disease recurrence. Twenty-five patients developed recurrence of their disease in the postoperative period, and 24 (96%) had reappearance of antigen in the urine 10.1 +/- 9.2 months prior to clinical evidence of recurrence. Of the 25 patients who remained disease free (mean follow-up, 48 months), 23 of 25 (92%) were urinary-antigen negative at last follow-up, although 8 patients had transient elevations of urinary TAA early in the postoperative period. Results indicate that postoperative measurement of urinary TAA in sarcoma patients permits selection of a subset of individuals at high risk for the development of recurrence. This knowledge can be of assistance in the postoperative management of these patients.

Adolescent↗
PubMed1984

Some simple multiple linear regression equations for estimation of maximal aerobic power in healthy Indian males.

An attempt has been made to evolve some simple multiple linear regression equations for the prediction of VO2 max from body weight, time for 3.2 km run and exercise dyspnoeic index ( DIstd Ex%). The predictor variables have been selected by examining the product moment correlations of body weight, relative body weight indices, time for 3.2 km run, chest expansion, height, and DIstd Ex% with VO2 max, based on data collected on 320 healthy Indian males (17-22 years). It has been observed that body weight, time for 3.2 km run and DIstd Ex% attained maximum correlations with VO2 max. Thus, two regression equations with two and three predictor variables have been established in this paper to predict VO2 max. The first regression equation yielded a multiple correlation of 0.608 (P less than 0.001) with a standard error of 0.214 1 X min-1. In this equation, body weight and time for 3.2 km run were considered as significant predictors. To increase the precision of this equation, another multiple linear regression equation based on body weight, time for 3.2 km run and DIstd Ex% as predictors has been developed. This equation yielded a multiple correlation of 0.658 (P less than 0.001) with a standard error of 0.204 1 X min-1. Applications of these regression equations will be of practical importance to biomedical scientists engaged in the development of a simple procedure for indirect assessment of VO2 max, and may serve well as preliminary screening procedures for personnel selection.

Adolescent↗