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Biomedical subjects

P C Sen

Publications and source records attributed to P C Sen.

At least 73 records · Page 4Linked to original sources

Characterization of partially purified (Na+ + K+)-ATPase from porcine lens.

The partial purification of (Na+ + K+)-ATPase from pig lens has been achieved by treatment with deoxycholate followed by density gradient centrifugation. The specific activity of the final preparation, ranging from 300 to 500 nmol/h per mg protein, is increased approx. 100-fold compared to the homogenate. A parallel increase in rho-nitrophenylphosphatase activity is also observed. Sodium dodecyl sulfate (SDS) gel electrophoresis reveals six major protein bands, one of which is the 93 kDa alpha subunit of (Na+ + K+)-ATPase which can be phosphorylated by reaction with [gamma-32P]ATP. A second band contains a glycoprotein which displays an apparent molecular weight of 51000 and thus appears to be the beta subunit of the enzyme. The enzyme is sensitive to ouabain with the I50 for (Na+ + K+)-ATPase and rho-nitrophenylphosphatase inhibition being 1.2 and 1.3 microM, respectively. Several agents which inhibit (Na+ + K+)-ATPase from other tissues such as oligomycin, Ca2+, vanadate, N-ethylmaleimide, rho-chloromercuribenzenesulfonic acid (PCMBS) and 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) also inhibit the lens enzyme. Monovalent cations other than K+ are partially effective in activating the (Na+ + K+)-ATPase and rho-nitrophenylphosphatase activities. The K+ congeners were relatively more effective in supporting (Na+ + K+)-ATPase compared to rho-nitrophenylphosphatase activity. Other kinetic properties of the lens enzyme are also comparable to those of the enzyme from other tissues. Utilizing the partially purified membrane bound enzyme, discontinuities in Arrhenius plots of (Na+ + K+)-ATPase activity, rho-nitrophenylphosphatase activity and fluorescence polarization of the fluidity probe, 1,6-diphenyl-1,3,5-hexatriene (DPH), are observed near the physiological temperature of lens. The possible significance of these observations for the mechanism of cataract formation are discussed.

4-Nitrophenylphosphatase↗

Studies of gastric Ca2+-stimulated adenosine triphosphatase. I. characterization and general properties.

Gastric microsomes do not contain any significant Ca2+-stimulated ATPase activity. Trypsinization of pig gastric microsomes in presence of ATP results in significant (2-3 fold) increase in the basal (with Mg2+ as the only cation) ATPase activity, with virtual elimination of the K+-stimulated component. Such treatment causes unmasking of latent Mg2+-dependent Ca2+-stimulation ATPase. Other divalent cations such as Sr2+, Ba2+, Zn2+, and Mn2+ were found ineffective as a substitute for Ca2+. Moreover, those divalent cations acted as inhibitors of the Ca2+-stimulated ATPase activity. The pH optimum of the enzyme is around 6.8. The enzyme has a Km of 70 microM for ATP and the Ka values for Mg2+ and Ca2+ are about 4 x 10(-4) and 10(-7) M, respectively. Studies with inhibitors suggest the involvement of sulfhydryl and primary amino groups in the operation of the enzyme. Possible roles of the enzyme in gastric H+ transport have been discussed.

Animals↗

Characterization of gastric-mucosal membranes. Distribution of lipid- and protein-associated amino groups across pig gastric microsomes.

Two NH2-reactive probes (2,4,6-trinitrobenzesulphonic acid and 1-fluoro-2,4-dinitrobenzene) were used to study the vectorial orientation of the membrane-associated free NH2 groups across pig gastric microsomal vesicles. Unlike 1-fluoro-2,4-dinitrobenzene, 2,4,6-trinitrobenzenesulphonic acid is ordinarily an impermeant probe that becomes permeant in the presence of K+ and valinomycin. Although 2,4,6-trinitrobenzenesulphonic acid alone reacts with about 28% of the total microsomal phosphatidylethanolamine, 2,4,6-trinitrobenzenesulphonic acid in the presence of valinomycin plus K+ or 1-fluoro-2,4-dinitrobenzene alone reacted with 75% of the phosphatidyl- ethanolamine. Under similar conditions the free NH2 groups associated with the microsomal proteins also exhibited an asymmetric labeling pattern, the intra- and extravesicular orientation being 74 and 26% respectively.

Animals↗

Immunological significance of lymphoreticular infiltration in gastrointestinal cancer.

The 60 patients of gastrointestinal cancer comprising this study were in the age range of 29--70 years with 39 males and 21 females. The primary sites of cancer were: gallbladder 19, colon 13, stomach 12, anorectum 8, and liver 8. The stage of tumor spread was localized in 32 and metastatic in 28 cases. Round cell infiltration scores were classified semiquantitatively from O-V. Cellular immunity as expressed by the blood lymphocyte count and DNCB reactivity was also studied. Round cell infiltration was dense in the localized group (P less than 0.0005). The mean lymphocyte counts in all the primary sites of gastrointestinal cancer were significantly higher in the localized group as compared to metastatic group. The DNCB reactivity showed anergy in six cases and poor response in the remaining 22 cases of metastatic group. There was a statistically significant correlation between lymphoreticular infiltration, peripheral blood lymphocyte count, and the DNCB reactivity. Diminished round cell infiltration, lymphopenia, and depressed DNCB response indicate an advanced stage of gastrointestinal cancer.

Adenocarcinoma↗

Characterization of gastric mucosal membranes. II. Lipid compositions of purified gastric mitochondria from pig, rabbit and frog.

1. The lipid composition of purified mitochondrial fractions from the fundic mucosa of pig, rabbit and frog were determined. 2. The total lipids expressed as mg lipid per 100 mg mitochondrial protein were approx. the same in pig and rabbit (13.4 and 15.5, respectively) and much higher than in frog (8.5). 3. The levels of phospholipids were about the same in pig and frog (approx. 61% of the total lipid) and lower than rabbit (78%). However, the levels of cholesterol were significantly different in the three species and constituted 22, 9 and 18.2% of the total lipids in pig, rabbit and frog mitochondria, respectively. 4. The glycolipid content in the mitochondrial lipids from pig, rabbit and frog were 7, 5.6 and 10.5%, respectively. 5. Cardiolipin contributed from 5.6 to 7.5% of the total phospholipids in the various species. Phosphatidylethanolamine and phosphatidylcholine together accounted for 80 90% of the total phospholipids in the various species; the contribution of phosphatidylcholine being always higher than that of phosphatidylethanolamine. Small but significant amounts of phosphatidylinositol were present in all species. 6. Generally, the predominant saturated fatty acid in the phospholipids was 16:0 from all species (except in phosphatidylethanolamine from pig and frog), and 18:1 and 18:2 were the predominant unsaturated fatty acids from all species. Sphingomyelin contained the highest amount of saturated fatty acids (over 80%) in both the species (pig and rabbit) studied.

Animals↗