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Biomedical subjects

N Singh

Publications and source records attributed to N Singh.

At least 541 records · Page 30Linked to original sources
PubMed1985

Yeast fatty acid synthase: structure to function relationship.

The yeast fatty acid synthase is a multifunctional enzyme composed of two nonidentical subunits in an alpha 6 beta 6 complex that is active in synthesizing fatty acids. The seven catalytic activities required for fatty acid synthesis are divided between the alpha and beta subunits such that the alpha 6 beta 6 complex has six complements of each activity. It has been proposed that these are organized into six centers for fatty acid synthesis. There are different opinions regarding the operation of these centers in the alpha 6 beta 6 complex, on view being that they are functionally independent and the other proposes half-sites activity for the complex. We have attempted to distinguish between these proposals by the most direct method of active site titration, i.e., quantitation of fatty acyl product in the absence of turnover. This was accomplished by using p-nitrophenyl thioacetate and thiophenyl malonate (in place of the coenzyme A analogues) as substrates along with NADPH, thereby depriving the yeast synthase of coenzyme A required to release product as fatty acyl coenzyme A. The amount of fatty acyl product formed was quantitated by gas-liquid chromatography, as well as by direct estimation of radioactivity in the product when p-nitrophenyl thio [1-14C] acetate was used as a substrate. In both cases, a stoichiometry of close to six was found for mole of fatty acid synthesized per mole of alpha 6 beta 6 complex. This indicates that there are six functional centers for fatty acid synthesis in the multifunctional yeast alpha 6 beta 6 fatty acid synthase and that these centers operate independently.(ABSTRACT TRUNCATED AT 250 WORDS)

Carbon Radioisotopes↗
PubMed1985

Poly ADP-ribosylation of histones in tumor promoter phorbol-12-myristate-13-acetate treated mouse embryo fibroblasts C3H10T1/2.

The tumor promoter phorbol-12-myristate-13-acetate (PMA) increases the poly ADP-ribosylation of acid extractable (0.2N H2SO4) nuclear proteins in mouse embryo fibroblasts C3H10T1/2. Catalase suppresses the reaction by approximately 50%. Polyacrylamide gel electrophoresis reveals that the core histones H2B, A24 and H3d serve as major poly ADP-ribose acceptors. Smaller amounts of poly ADP-ribose are associated with histones H2A/H3 and H1. Poly ADP-ribosylation of histones may change the nucleosomal structure and function and play a role in PMA induced modulation of gene expression in promotion.

Animals↗
PubMed1985

The development and application of a novel chromophoric substrate for investigation of the mechanism of yeast fatty acid synthase.

The acetyl transacylase activity of the fatty acid synthase from yeast has been investigated using p-nitrophenylthiol acetate. The chromophoric nature of the nitrophenylthiol moiety affords a convenient spectrophotometric assay for the transacylase function as well as a means to investigate the kinetics and the mechanism of this process. A probable kinetic scheme for enzyme catalyzed transacetylation from p-nitrophenylthiol acetate to an acyl acceptor (CoA or N-acetylcysteamine) is proposed and the kinetic constants for acetylation of enzyme and for acetyl transfer to an acceptor were determined. It was also demonstrated that p-nitrophenylthiol acetate can replace acetyl-CoA as a substrate in fatty acid synthesis.

Acetyl Coenzyme A↗
PubMed1985

Tumor promoter phorbol-12-myristate-13-acetate induces poly ADP-ribosylation in human monocytes.

The tumor promoter phorbol-12-myristate-13-acetate (PMA) induces rapid poly ADP-ribosylation and a drop in cellular NAD concentration in human monocytes. The antioxidants CuZn-superoxide dismutase, catalase, glutathione peroxidase and butylated-hydroxytoluene inhibit the reaction indicating that active oxygen species produced in the PMA-induced oxidative burst represent intermediates. The inhibitor of ADP-ribosyl-transferase, 3-amino-benzamide, inhibited poly ADP-ribosylation but did not prevent the drop in NAD-levels. PMA also causes the slow accumulation of DNA strand breaks in monocytes. The difference in the kinetics of poly ADP-ribosylation and DNA breakage argues against a simple relationship between the two reactions.

Antioxidants↗
PubMed1985

Tumor promoter phorbol-12-myristate-13-acetate induces poly(ADP)-ribosylation in fibroblasts.

The tumor promoter phorbol-12-myristate-13-acetate (PMA) causes an increase in pol(ADP)-ribosylation in mouse and human fibroblasts via the intermediate formation of active oxygen. In contrast to poly(ADP)-ribosylation induced by the methylating agent N-methyl-N'-nitro-N-nitrosoguanidine, de novo RNA and protein synthesis are required and the accumulation of the polymer occurs in the absence of detectable DNA strand breakage. Our results suggest a mechanism for PMA-induced modulation of chromatin structure and gene expression.

Animals↗
PubMed1985

Non-histone chromosomal protein acceptors for poly(ADP)-ribose in phorbol-12-myristate-13-acetate treated mouse embryo fibroblasts (C3H10T1/2).

The tumor promoter phorbol-12-myristate-13-acetate (PMA) increases the level of poly ADP-ribosylation of chromosomal proteins in mouse embryo fibroblasts C3H10T1/2. The poly ADP-ribosylated nuclear proteins fall into the following molecular weight classes: 40, 48, 61, 77, 92, 158, 200 kd. Preincubation with catalase reduced the poly ADP-ribose (ADPR) substitution of all these proteins essentially to control levels. Western blot analysis with antibody directed against ADPR transferase indicates that the major acceptors are ADP-ribose transferase (116 kd) itself and its proteolytic degradation products of 20-25, 45 and 72-95 kd. Poly ADP-ribosylation of these proteins is suppressed by cycloheximide, 3-aminobenzamide, antipain and catalase. The latter three inhibitors possess anti-promotional activities in certain in vitro cell culture systems. Auto-poly ADP-ribosylation of ADPR transferase and its proteolytic cleavage as well as the poly ADP-ribosylation of other chromosomal proteins may play a role in the modulation of gene expression by PMA.

Animals↗
PubMed1985

Evidence of immunity induced by naturally acquired rotavirus and Norwalk virus infection on two remote Panamanian islands.

For better understanding of the role of humoral immunity in ameliorating infections with rotavirus (RV) and Norwalk virus (NW), 305 Cuna Indians living on two isolated islands located off Panama's Carribean coast were surveyed daily for diarrhea over a seven-month period. Nine (8%) of 108 persons with a baseline RV antibody titer of greater than 1:4 developed RV infection compared with 70 (46%) of 151 persons with a baseline RV antibody titer of less than 1:4 (P less than .001). Thirty-eight (25%) of 151 persons of all ages with baseline RV antibody titer of less than 1:4 had at least one episode of RV diarrhea compared with 6 (6%) of 108 persons who had baseline RV antibody titers of greater than 1:4 (P less than .001). Thirty-two (47%) of 68 persons of all ages who had a baseline NW antibody titer of less than 1:100 developed NW infection compared with 30 (13%) of 237 persons with a baseline NW titer of greater than 1:100 (P less than .001). The high NW and RV infection rates and the excellent levels of protection provided by specific preexisting humoral antibody to these agents should promote activities aimed at developing vaccines for preventing these infections.

Age Factors↗
PubMed1985

Norwalk virus gastroenteritis. An outbreak associated with a cafeteria at a college.

An explosive outbreak of gastrointestinal illness occurred among students and employees at a small college in Florida in November 1980. Common symptoms were diarrhea, nausea, weakness, abdominal cramps, chills, vomiting, and low-grade fever. Cases of illness were identified in 40% of 628 students and 15% of 162 employees who responded to a survey. Among students, there was a sevenfold excess risk associated with eating one or more meals at the campus cafeteria November 3-5 (p much less than 0.001). Tossed salad from one meal was strongly associated with illness (p less than 0.0001). Fecal contamination of the salad was documented, although the source of contamination was not identified. Person-to-person spread could not be demonstrated. Seroconversion to Norwalk antigen occurred in significantly more cases (5/6) than noncases (1/6) (p = 0.04).

Disease Outbreaks↗
PubMed1985

The childhood health effects of an improved water supply system on a remote Panamanian island.

The incidence of diarrhea, respiratory disease, and skin infections was prospectively determined after the introduction of a system which distributed unlimited quantities of high quality fresh water to each of the 150 housing units on Tupile, an island devoid of fresh water located off Panama's Caribbean coast and inhabited by 1,500 Cuna Indians. Tupile residents used 7.1 liters of water/person/day compared to the 2.3 usage rate of inhabitants on Achutupo, the control island. Despite ready availability of water in each household, Tupile residents continued to store water in contaminated vessels prior to use. Forty percent of stored water samples tested on Tupile and 45% on Achutupo were contaminated with E. coli organisms. There were 4.7 episodes/child year (E/Y) of acute diarrhea on Tupile compared with the 3.5 rate on Achutupo. The rotavirus infection rate on Tupile was 0.8 E/Y compared with 0.2 E/Y on Achutupo. Infection rates for Norwalk virus, respiratory syncytial virus and Coxsackie B 1-6 viruses were similar on both islands. Respiratory disease rates were high on both islands (2.2 E/Y on Tupile, 2.7 E/Y on Achutupo). Achutupo had much higher rates of impetigo and scabies (0.6 E/Y and 2.5 E/Y, respectively) than Tupile (0.2 E/Y and 1.4 E/Y). Provision of the water distribution system had a beneficial effect on the incidence of water-washed diseases (impetigo and scabies), but at best had no effect on diarrheal disease.

Child↗
PubMed1985

Scleromyxedema.

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Glycosaminoglycans↗