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Biomedical subjects

N Singh

Publications and source records attributed to N Singh.

At least 487 records · Page 27Linked to original sources
PubMed1991

Two different mutants blocked in synthesis of dolichol-phosphoryl-mannose do not add glycophospholipid anchors to membrane proteins: quantitative correction of the phenotype of a CHO cell mutant with tunicamycin.

The addition of glycophospholipid (GPL) anchors to certain membrane proteins occurs in the rough endoplasmic reticulum and is essential for transport of the proteins to the plasma membrane. Limited circumstantial evidence suggests that dolichol-phosphoryl-mannose (DPM) is a donor of mannose residues of these anchors. We here report studies of a CHO cell mutant (B421) transfected to express the GPL-anchored protein, placental alkaline phosphatase (AP). Only a few transfectants were found to express GPL-anchored AP on their surface, and these clones synthesized DPM. Moreover, and most strikingly, when surface AP-negative transfectants were treated with tunicamycin to cause accumulation of DPM, these cells expressed lipid-anchored AP. Fusion of a cloned surface AP-negative transfectant of B421 with the Thy-1-class E mutant thymoma, which is also deficient in DPM synthesis, produced hybrids that synthesized DPM and expressed AP and Thy-1. Thus, two mutations can interrupt DPM synthesis, and three sets of observations point to an essential role of DPM for addition of GPL anchors.

Alkaline Phosphatase↗
PubMed1991

Anchoring and degradation of glycolipid-anchored membrane proteins by L929 versus by LM-TK- mouse fibroblasts: implications for anchor biosynthesis.

Although many cells anchor surface proteins via moieties that are sensitive to phosphatidylinositol-specific phospholipase C (PI-PLC), the anchor moieties of surface proteins of mouse L929 cells resist PI-PLC. By constructing stable hybrids between L929 and lymphoma cells that express glycolipid-anchored proteins in a PI-PLC-sensitive form, we show that PI-PLC resistance behaves as a recessive trait. Since putative mannolipid precursors of the lipid anchors bear alkali-labile substituents which make them resist PI-PLC, these observations are most simply interpreted by postulating that L929 lacks a critical anchor deacylase. Unlike the L929 cell line, two of its descendants, the LM cell line and its thymidine kinase-negative variant (LM-TK-), do not express glycolipid-anchored proteins on their surface. Moreover, unlike L929 cells, LM-TK- cells rapidly inactivate at least one lipid-anchored enzyme in a compartment sensitive to acidotropic amines and leupeptin. By fusion of LM-TK- cells to mouse Thy-1- lymphoma mutants and monitoring of surface expression of lipid-anchored proteins, we assign LM-TK- to lymphoma mutant complementation group H. This genetic assignment is matched by analysis of mannolipids of L929, LM-TK-, wild-type, and class H lymphoma mutant cells: striking similarities are seen between the two wild-type cells by contrast to the mutants. Since the differences pertain to lipids which have properties consistent with their being anchor precursors, we suggest that LM-TK- has a lesion in the synthesis of anchor precursor mannolipids.

Alkaline Phosphatase↗
PubMed1991

Response of extrapyramidal and limbic neuropeptides to fenfluramine administration: comparison with methamphetamine.

The responses of extrapyramidal and limbic neuropeptide and striatal dopamine and serotonin systems were evaluated after treatment with fenfluramine in rats. After multiple administrations of fenfluramine, its active metabolite, norfenfluramine, and methamphetamine (METH), striatal neurotensin (NT) content was similarly increased to approximately 200% of control. In contrast, nigral NT levels were unaltered by fenfluramine, intermediately increased by norfenfluramine (148% of control) and maximally increased by METH (267% of control). Striatal and nigral substance P (SP) and dynorphin A (Dyn) systems were unaltered by fenfluramine, whereas norfenfluramine caused an intermediate increase in striatal Dyn content but did not significantly alter striatal SP or nigral SP and Dyn levels. However, METH significantly elevated striatal and nigral Dyn and SP concentrations to 280 to 425% (Dyn) and 140% (SP) of control. For the most part, the response of the limbic peptides was similar to that seen in the striatum with a couple of notable differences. Further investigation of the striatal NT system showed that the increases induced by fenfluramine were completely blocked by the D1 antagonist, SCH 23390, and the noncompetitive N-methyl-D-aspartate antagonist, MK801. Depletion of 5-hydroxytryptamine with pretreatment by parachloroamphetamine did not alter the response of the striatal NT system to fenfluramine. The present results demonstrate common and unique features in the response of peptide systems to fenfluramine and methamphetamine, which might explain some of the similarities and differences between these two drugs.

Animals↗
PubMed1991

Delayed mortality and morphogenetic anomalies induced in Culex quinquefasciatus by the microbial control agent Bacillus sphaericus.

Two preparations of Bacillus sphaericus 2362 were studied for their biological activity, delayed mortality and the induction of morphogenetic aberrations in larvae, pupae and adults of Culex quinquefasciatus. Longevity and fecundity of adult mosquitoes were also assessed. A dosage response line for B. sphaericus was established against 4th-instar larvae and sublethal concentrations (48 h LC50 and lower) were used against these larvae. Sublethal concentrations of B. sphaericus induced delayed mortality in larvae, pupae and adults. The magnitude of mortality increased in succeeding cohorts and developmental stages resulting from the surviving larvae. Only 10 and 25% overall emergence of viable adults occurred in the sublethal treatments (LC25) of 2 B. sphaericus preparations. The range of successful adult emergence was over 94% in the controls. A wide range of external morphogenetic aberrations in dead larvae, pupae and adults were noted. These aberrations and gross morphological features were quite similar to those reported for certain insect growth regulators. Sublethal concentrations had no marked effect on longevity of adults, egg deposition and hatch.

Abnormalities, Multiple↗
PubMed1991

Delayed mortality and morphogenetic anomalies induced by the microbial control agent Bacillus thuringiensis ser. (H-14) in Culex quinquefasciatus.

Sublethal concentrations of Bacillus thuringiensis ser. H-14 were applied to early 4th-instar larvae of Culex quinquefasciatus to assess mortality and morphogenetic aberrations in larvae, pupae and adults. At the 24 h LC10, LC25, LC50 and LC80, additional mortality occurred in surviving larvae beyond a 24 h exposure period. The cumulative mortalities increased daily and the overall mortality of larvae up to 7 days posttreatment were 12, 73, 82 and 96% at the indicated concentrations, respectively. Delayed mortality also occurred in the pupal and adult stages. Morphogenetic aberrations were noted in dead larvae and pupae but were rare in the adults. These aberrations are categorized and described. There was little or no delayed effect on the survivorship or fecundity of adults, but in all the treatments the number of emerging males was higher than females. The sex ratio in check adults was 1:1.

Animals↗
PubMed1990

A comparative study on the effect of tumor promoters on poly ADP-ribosylation in A431 cells.

Poly ADP-ribosylation is a post-translational modification of chromatin proteins catalyzed by the enzyme poly-ADPR transferase (poly-ADPRT) and affects the structure as well as the functional properties of chromatin. It is of particular relevance in carcinogenesis, as it represents an epigenetic mechanism for modulation of gene expression. In the present study, A431 cells were exposed to tumor promoters phorbol-12-myristate-13-acetate (PMA), benzoyl peroxide (BP), mezerein and 6-keto-lithocholic acid (KA), and their effect on poly-ADP-ribosylation was studied. All these tumor promoters increased the activity of poly-ADPRT in these cells--PMA 2.3-fold, BP and mezerein 2.2-fold each and KA 1.3-fold. The enzyme inhibitor 3-amino benzamide (3AB) partially prevented the stimulation of poly-ADPRT by these promoters. There was a concomitant decrease in NAD levels, the substrate for poly-ADPRT. The decrease was 44% for PMA, 46% for BP, 21% for KA and 34% for mezerein. The induction of poly-ADP-ribose synthesis by PMA and BP appears to be mediated at least in part by active oxygen species, as they induced an increase in superoxide anions and anti-oxidants prevented the increase of poly-ADPRT activity to varying extents.

Benzamides↗
PubMed1990

The yeast fatty acid synthase. Pathway for transfer of the acetyl group from coenzyme A to the Cys-SH of the condensation site.

The reaction pathway of enzyme-catalyzed acetylation of the acyl-accepting sites of the yeast synthase, a Ser-OH at the acetyl transacylase site, a Cys-SH at the beta-ketoacyl synthase site, and the acyl carrier protein 4'-phosphopantetheine-SH (Pant-SH), has been investigated using the chromophoric substrate, p-nitrophenyl thioacetate. The stoichiometry of acetylation of the native enzyme was 3 mol of acetate bound per mol of synthase unit, alpha beta (Mr 430,000). The acetylation process is biexponential; the rate constant of acetylation of the first 2 mol is 5.0 s-1 and the third mol is 0.2 s-1. The pathway by which acetyl moiety is added to the enzyme was determined by selectively blocking the acyl-accepting sites and subsequently determining the kinetics and stoichiometry of acetylation. The dibromopropanone-treated enzyme, in which the Pant-SH and Cys-SH are alkylated, exhibited an exponential burst of approximately 1 mol/mol of synthase unit with a rate constant of 11.0 s-1. The iodoacetamide-treated enzyme, in which Cys-SH is alkylated, had a biexponential burst with a total stoichiometry of approximately 2 mol/mol of synthase unit, with rate constants of 9 and 0.2 s-1, respectively. The kinetically competent acetylation to the extent of 2 and approximately 1 mol/mol of synthase unit for both Cys-SH and Cys-SH and Pant-SH-blocked enzymes, respectively, indicated that the route of acetyl transfer in the yeast synthase is obligatorily Ser-OH----Cys-SH. The acetylation of Pant-SH (0.2 s-1) occurs with a rate insignificant to the process of fatty acid synthesis (turnover rate constant of 1.5 s-1). These conclusions are supported by experiments involving end point radiolabeling of the synthase with [1-14C]acetyl moieties using the substrate, p-nitrophenyl thio[1-14C]acetate. Native, dibromopropanone-treated, and iodoacetamide-treated enzymes bind about 3, 1, and 2 mol of acetyl/mol of synthase unit, respectively. Performic acid oxidation studies of the acetyl-labeled enzyme indicate that there is one Ser-O-acetyl formed in the native and alkylated enzymes and one Cys-S-acetyl and one Pant-S-acetyl formed in the native enzyme. Altogether, these results support our contention that the acetylation of the Pant-SH is kinetically incompetent. Thus, the yeast synthase transacetylation reactions occur by a novel process of acetyl transfer from CoA to Ser-OH----Cys-SH, which is in contrast to the transfer from CoA to Ser-OH----Pant-SH----Cys-SH catalyzed by the prokaryotic synthases.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylation↗
PubMed1990

Complete and partial glycophospholipid anchors are found on a fusion protein consisting of luteinizing hormone beta subunit followed by a carboxyl-terminal domain of Thy-1.

The Thy-1 antigen is anchored to the cell surface by a carboxyl-terminal glycophospholipid moiety. To investigate the extent of anchor addition which occurs when such proteins cannot move efficiently to the cell surface, we have expressed a recombinant fusion protein composed of 107 amino-terminal amino acids of bovine luteinizing hormone beta subunit and 46 COOH-terminal amino acids of murine Thy-1 (Thy-1.2 allele). Although the limited amount of fusion protein transported to the cell surface is glycophospholipid-anchored, most of the protein accumulates in an intracellular, endoglycosidase H-sensitive form. The intracellular protein has an unusual structure that contains ethanolamine but does not bind detergent, suggesting either that anchor addition proceeds via a hydrophilic partial intermediate, or that anchor-degradative enzymes exist along the secretory path.

Animals↗
PubMed1990

Effect of uterine fluid on in vitro acrosome reaction of buffalo (Bubalus bubalis ) spermatozoa.

The present study was undertaken to examine in vitro acrosome reaction in the uterine fluid of estrous buffalo. Successful acrosome reaction was achieved by incubating buffalo spermatozoa in 2% detoxified uterine fluid in Biggers Whitten Whittingham (BWW) medium, pH 7.4 at 37 degrees C and a sperm concentration of 36 x 10/ml. Neat uterine fluid has been found to be toxic to spermatozoa, hence we detoxified the uterine fluid at 56 degrees C for 30 min, which resulted in higher percentage of sperm motility, viability, and acrosome reaction. All the three stages of acrosome reaction i.e., acrosome swelling, vesiculation and shedding, were observed and they reached an apparent maximum at 4, 7 and 8 h of incubation, respectively. The significance of the findings in relation to the role of female reproductive tract in acrosome reaction is discussed.

Journal Article↗
PubMed1990

Effect of cement dust on the growth and yield of Brassica campestris L.

Plants of Brassica campestris L. var. G-S20 were treated with cement dust, at rates of 3 (B(1)), 5 (B(2)) and 7 (B(3)) gm(-2) day(-1) for 90 days. Treated plants showed a consistent reduction in growth, photosynthetic pigments, yield and oil content over control plants. The overall phytomass of treated plants was significantly decreased, the maximum reduction being 64.8% in B(3) plants, followed by B(2) plants (55.3%) and B(1) plants (43.69%) at 60 days. The effect on oil content was also greatest in B(3) plants, where it was decreased by 6.13%.

Journal Article↗
PubMed1990

Haloperidol as a pre-medicant for thiopental anaesthesia in the dog.

Haloperidol was administered intravenously at the dose rate of 0.87 mg/kg body weight five minutes prior to thiopental anaesthesia in 5 clinically healthy dogs, aged 10-12 months and weighing 11.5 +/- 0.96 kg. Animals required only 4.36 +/- 0.24 ml of the 5% thiopental sodium to achieve surgical anaesthesia which lasted for 37.5 +/- 4.3 minutes. There was adequate muscle relaxation and loss of pedal and palpebral reflexes during thiopental anaesthesia. Five minutes after administration of haloperidol, there was no appreciable change in the various cardiopulmonary dynamics with the exception of a hypocapnoea and a mild hypotension. During thiopentone anaesthesia, a mild hypotension and arterial hypoxemia was evident. This combination of anaesthesia was also employed in 14 clinical cases varying from fractures of long bones (4), mammary tumours (3), ear haematoma (4), venereal granuloma (2) and abdominal hernia (1). The combination proved extremely useful for orthopaedic surgery as the muscle relaxation was adequate and the reduction of the fractured ends was relatively easy.

Anesthesia↗