xenobiotic-metabolizing enzyme cyp2a5: Topics by WorldWideScience.org

Transposable elements are enriched within or in close proximity to xenobiotic-metabolizing cytochrome P450 genes

Rats or mice acutely exposed to high concentrations of ozone show an immediate and significant weight loss, even when allowed free access to food and water. The mechanisms underlying this systemic response to ozone have not been previously elucidated. We have applied the technique of global gene expression analysis to the livers of C57BL mice acutely exposed to ozone. Mice lost up to 14% of their original body weight, with a 42% decrease in total food consumption. We previously had found significant up-regulation of genes encoding proliferative enzymes, proteins related to acute phase reactions and cytoskeletal functions, and other biomarkers of a cachexia-like inflammatory state in lungs of mice exposed to ozone. These results are consistent with a general up-regulation of different gene families responsive to NF-#kappa#B in the lungs of the exposed mice. In the present study, we observed significant down-regulation of different families of mRNAs in the livers of ...

2005-10-15

3

Cobalamin- and Corrinoid-Dependent Enzymes

BackgroundTransposons, i.e. transposable elements (TEs), are the major internal spontaneous mutation agents for the variability of eukaryotic genomes. To address the general issue...Full Text Available

4

Polymer materials for enzyme immobilization and their application in bioreactors

This chapter will review the literature on cobalamin- and corrinoid-containing enzymes. These enzymes fall into two broad classes, those using methylcobalamin or related methylcorrinoids as...Full Text Available

2009-01-01

5

Effect of Spices on hepatic microsomal enzyme function in mice

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2011-02-01

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Display of Recombinant Proteins on Bacillus subtilis Spores, Using a Coat-Associated Enzyme as the Carrier?

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1984-06-01

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Chemical complementation: A reaction-independent genetic assay for enzyme catalysis

The display of proteins such as feed enzymes at the surface of bacterial spore systems has a great potential use for animal feed. Feed enzymes increase the digestibility of nutrients, leading to greater...Full Text Available

2010-09-01

8

The kinetic model of the shikimate pathway as a tool to optimize enzyme assays for high-throughput screening.

A high-throughput assay for enzyme activity has been developed that is reaction independent. In this assay, a small-molecule yeast three-hybrid system is used to link enzyme catalysis to transcription...Full Text Available

2002-12-24

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9902631 - Lysozyme - NASA

Four-enzyme section of the shikimate pathway (Aro B, D, E, and K) of Streptococcus pneumoniae has been studied. Kinetic properties of the individual enzymes and three- and four-enzyme linked reactions have been characterized in vitro. On the basis of the data measured in spectrophotometric and LC-MS experiments, kinetic mechanisms of the enzymes have been suggested and all kinetic parameters have been identified. Kinetic models for these three- and four-enzyme sections of the shikimate pathway have been constructed and validated. The model of the four-enzyme section of shikimate pathway has been employed to design an inhibition-sensitive reconstituted pathway for a high-throughput screening effort on the shikimate pathway. It was demonstrated that using the model it was possible to optimize this reconstituted pathway in such a way to provide equal sensitivity of ...

2006-11-01

10

Secondary Structure Analysis of a G-rich Sequence Recognizing RNA Aptamer with Structure Specific Enzymes

Protein isolated from hen egg-white and functions as a bacteriostatic enzyme by degrading bacterial cell walls. First enzyme ever characterized by protein ...

11

Purification and Characterization of Angiotensin I-Converting Enzyme Inhibitors from Sinapis alba L.

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2011-06-01

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Protective effects of Opuntia ficus-indica on Glutathione and Antioxidative Enzyme

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2000-06-01

13

Meta Analysis to Draw the Appropriate Regimen of Enzyme and Probiotic Supplementation to Pigs and Chicken Diets

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2003-12-01

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Loss of contraction force in dermal fibroblasts with aging due to decreases in myosin light chain phosphorylation enzymes

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2011-04-01

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Isolation and Characterization of Acinetobacter sp. BD5 Producing Lipolytic Enzyme

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Enzymic synthesis of 1-O-(indol-3-ylacetyl)-beta-D-glucose. Purification of the enzyme from Zea mays, and preparation of antibodies to the enzyme.

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2006-07-01

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Effect of simulated Acid Rain on the Morphology and Enzyme of Perilla frutescens var. japonica Hara

The enzyme indol-3-ylacetylglucose synthase (UDP-glucose:indol-3-ylacetate beta-D-glucosyltransferase) catalyses the reaction: [formula: see text] This is the first step in the series of reactions leading...Full Text Available

1991-10-15

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Effect of Di-(2-ethylhexyl)-phthalate on Sphingolipid Metabolic Enzymes in Rat Liver

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1996-04-01

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Effect of Cyclohexanone Treatment on the Activities of Oxygen Free Radical Metabolizing Enzyme in the Liver Damaged Rats

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2011-09-01

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Effect of Circadian Rhythms on the Xylene Metabolizing Enzyme Activities in Rats

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2002-06-01

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Effect of Angiotensin-I Converting Enzyme Inhibitory from Hydrolysate of Soybean Protein Isolate

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2001-06-01

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Changes in Antioxidant Enzyme Activities in the Gill And Digestive Glands of the Manila Clam Ruditapes philippinarum exposed to Cu

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2010-01-01

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Antioxidant and Angiotensin Converting Enzyme I Inhibitory Activity on Different Parts of Germinated Rough Rice

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2010-03-01

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Unexpected diversity of RNase P, an ancient tRNA processing enzyme: challenges and prospects

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2011-06-01

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Proteinases and Enzyme Stability in Crude Extracts of Castor Bean Endosperm ¹

For an enzyme functioning predominantly in a seemingly housekeeping role of 5′ tRNA maturation, RNase P displays a remarkable diversity in subunit make-up across the three domains of...Full Text Available

2010-01-21

26

Phosphoglycerate kinase and triosephosphate isomerase from the hyperthermophilic bacterium Thermotoga maritima form a covalent bifunctional enzyme complex.

The stability of catalase, fumarase, and isocitrate lyase from deliberately broken organelles in crude extracts from endosperm tissue of castor bean seedlings has been examined. These enzymes are relatively...Full Text Available

1981-03-01

27

Partial Purification and Characterization of Indol-3-Ylacetylglucosemyo-Inositol Indol-3-Ylacetyltransferase (Indoleacetic Acid-Inositol Synthase) ¹

Phosphoglycerate kinase (PGK) from the hyperthermophilic bacterium Thermotoga maritima has been purified to homogeneity. A second larger enzyme with PGK activity and identical N-terminal sequence was...Full Text Available

1995-02-01

28

Overexpression of bacterial ethylene-forming enzyme gene in Trichoderma reesei enhanced the production of ethylene

A procedure is described for the purification of the enzyme indol-3-ylacetylglucose:myo-inositol indol-3-ylacetyltransferase (IAA-myo-inositol synthase). This enzyme...Full Text Available

1990-12-01

29

Evaluation of the microparticle enzyme immunoassay Abbott IMx Select Chlamydia and the importance of urethral site sampling to detect Chlamydia trachomatis in women.

In order to efficiently utilize natural cellulose materials to produce ethylene, three expression vectors containing the ethylene-forming enzyme (efe) gene from Pseudomonas...Full Text Available

30

Enzyme-Linked Immunosorbent Assay for Recombinant K39 Antigen in Diagnosis and Prognosis of Indian Visceral Leishmaniasis

OBJECTIVE: To evaluate the commercial microparticle enzyme immunoassay (MEIA), Abbott IMx Select Chlamydia, for the detection of Chlamydia trachomatis in women and to compare its performance with endocervical...Full Text Available

1997-12-01

31

Enzyme amplified immunoassay: a novel technique applied to direct detection of Chlamydia trachomatis in clinical specimens.

The recombinant product (rK39) of the 39-amino-acid repeats encoded by a kinesin-like protein-encoding gene of Leishmania chagasi was evaluated by enzyme-linked immunosorbent assay...Full Text Available

2001-11-01

32

Efficient preparation of internally modified single-molecule constructs using nicking enzymes

Endocervical swabs from 212 women and urethral swabs from 100 men were tested by the routine methods for McCoy cell culture and simultaneously by a novel enzyme amplified immunoassay test to detect...Full Text Available

1985-10-01

33

Determination of antibody response to influenza virus surface glycoproteins by kinetic enzyme-linked immunosorbent assay.

Investigations of enzymes involved in DNA metabolism have strongly benefited from the establishment of single molecule techniques. These experiments frequently require elaborate DNA substrates, which...Full Text Available

2011-02-01

34

Cellulase-Xylanase Synergy in Designer Cellulosomes for Enhanced Degradation of a Complex Cellulosic Substrate

We modified an existing enzyme-linked immunosorbent assay (ELISA) to be able to use new spectrophotometers which can measure the rate of color development in microtiter wells. This new kinetic-based...Full Text Available

1988-10-01

35

CDC45 and DPB11 are required for processive DNA replication and resistance to DNA topoisomerase I-mediated DNA damage

Designer cellulosomes are precision-engineered multienzyme complexes in which the molecular architecture and enzyme content are exquisitely controlled. This system was used to examine enzyme cooperation...Full Text Available

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Biotransformation of Explosives by the Old Yellow Enzyme Family of Flavoproteins

The antitumor agent camptothecin targets DNA topoisomerase I by reversibly stabilizing a covalent enzyme-DNA intermediate. The subsequent collision of DNA replication forks with these drug-enzyme-DNA...Full Text Available

1999-09-28

37

Beta-lactamases as fully efficient enzymes. Determination of all the rate constants in the acyl-enzyme mechanism.

Several independent studies of bacterial degradation of nitrate ester explosives have demonstrated the involvement of flavin-dependent oxidoreductases related to the old yellow enzyme (OYE) of yeast....Full Text Available

2004-06-01

38

Allosteric regulation of glycerol kinase by enzyme IIIglc of the phosphotransferase system in Escherichia coli and Salmonella typhimurium.

The rate constants for both acylation and deacylation of beta-lactamase PC1 from Staphylococcus aureus and the RTEM beta-lactamase from Escherichia coli were determined by the acid-quench method [Martin...Full Text Available

1990-03-15

39

Activation of cycasin to a mutagen for Saccharomyces cerevisiae by rat intestinal flora.

The mechanism by which enzyme IIIglc of the bacterial phosphotransferase system regulates the activity of crystalline glycerol kinase from Escherichia coli has been studied, and the inhibitory effects...Full Text Available

1985-05-01

40

A rationale for stabilization of oxygen-labile enzymes: application to a clostridial hydrogenase.

Genetic test systems involving microorganisms and liver enzyme preparations may be insufficient to detect compounds that require breakdown by enzymes provided by the microbial flora of the intestinal...Full Text Available

1983-02-01

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Excitation Energy Transfer Study of the Spatial Relationship ...

A general procedure for stabilization of O2-labile enzymes exploiting "salting out" of oxygen from the microenvironment in the molecular layers immediately adjacent to charged surfaces of polyionic...Full Text Available

1978-08-01

42

Effects of Green Tea Catechin on Serum Lipid Composition and Angiotensin- Converting Enzyme Activity in Rats Exposed to Microwave

... closely correlated to the amount of re- centration. The molar extinction coefficients were agent added to the enzyme. The naphtha- ...

1988-02-15

43

Effects of Feeding the Mixed Oils of Butter, Sardine and Safflower on the Lipid Components in Serum and Activities of Hepatic Functional Enzymes in Rats

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2004-06-01

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Effect of Medicinal Plant Water Extracts on Glucose-regulating Enzyme Activities in Goto-Kakizaki Rat Liver Cytosol

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1992-12-01

45

"6"7Ga scintigraphy, serum lysozyme and angiotensin-converting enzyme in pulmonary sarcoidosis

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2009-10-01

46

Enzyme discovery in hybrid aspen for fibre engineering

... angiotensin gallium 67 lungs lysozyme patients sarcomas beta decay

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Environmental Research Database

DescriptionEnzymes are natural catalysts that are used increasingly to replace environmentally harmful chemicals and to achieve better catalytic selectivity during pulp and paper processing. The main objective of this project is to identify novel plant enzymes for deeper understanding of the process of fibre formation and for future improvement of the quality parameters of wood fibres. Specific modification of the fibre characteristics will facilitate their use as raw materials for the current pulp and pap [continued...

2005-01-31

48

Lysis of Escherichia coli cells by lysozyme: Discrimination between adsorption and enzyme action

British Library Electronic Table of Contents (United Kingdom)

The key factors of enzymatic lysis of cells are the interaction between the enzyme and the cell - catalytic and non-catalytic adsorption of enzyme on cell surface. Here, the studies of lysis of intact Escherichia coli cells by chicken egg white lysozyme were performed. It was found that the ionic strength has a dual effect onto the system. On the one hand, the desorption constant of the enzyme increases with the increase of the solution ionic strength, which results in a better enzyme performance. On the other hand, due to the higher osmosis, the cell lysis rate decreases with the increasing of ionic strength of the system. It was found that pH 8.6 and 30mM NaCl are optimal conditions for lysis of E. coli cells by lysozyme.

2011-01-01

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Lipase-mediated hydrolysis of corn DDGS oil: Kinetics of linoleic acid production

British Library Electronic Table of Contents (United Kingdom)

In this study, we investigated the kinetics of linoleic acid production via lipase-mediated hydrolysis of corn DDGS oil in a batch reactor with continuous mechanical agitation and developed a kinetic model that incorporated the product inhibition to study the complete hydrolysis. The model agreed very well with observed data; though situations with low enzyme dosage or low stirring rates were modeled successfully without product inhibition, actual product concentration in such situations was too low to exert any inhibitory effects. Increasing the enzyme concentration increased hydrolysis, and beyond certain enzyme concentrations, effects tended to fade away because of excessive enzyme desorption from the interface. An enzyme dosage within the range of 40-60KLU/L of oil dispersion could be ...

2010-01-01

50

Distinguishing two groups of flavin reductases by analyzing the protonation state of an active site carboxylic acid

British Library Electronic Table of Contents (United Kingdom)

Abstract Flavin-containing reductases are involved in a wide variety of physiological reactions such as photosynthesis, nitric oxide synthesis, and detoxification of foreign compounds, including therapeutic drugs. Ferredoxin-NADP(H)-reductase (FNR) is the prototypical enzyme of this family. The fold of this protein is highly conserved and occurs as one domain of several multidomain enzymes such as the members of the diflavin reductase family. The enzymes of this family have emerged as fusion of a FNR and a flavodoxin. Although the active sites of these enzymes are very similar, different enzymes function in opposite directions, that is, some reduce oxidized nicotinamide adenine dinucleotide phosphate (NADP+) and some oxidize reduced nicotinamide adenine dinucleotide phosphate (NADPH). In t...

2011-01-01

51

A Comparative Biochemical Characterization of Microbial Transglutaminases: Commercial vs. a Newly Isolated Enzyme from Streptomyces Sp.

British Library Electronic Table of Contents (United Kingdom)

A new microbial transglutaminase (MTGase or MTG, EC 2.3.2.13) from a Streptomyces sp. strain isolated from Brazilian soil samples was characterized in crude and purified forms. The aim of this work is to provide relevant information about a new transglutaminase and to compare its characteristics with the well-known commercial transglutaminase from Ajinomoto Co. Inc. (Activa? TG-BP). The enzyme from Streptomyces sp., in both crude and pure forms, exhibited optimal activity in the 6.0?6.5 pH range and at 35?40?C. The results for the commercial enzyme were the same. A second maximum of activity was observed at pH?10.0 with both the crude Streptomyces sp. enzyme and the commercial enzyme. This interesting fact has not been reported in the literature previously. The fact that this second maximu...

2010-01-01

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Yeast allosteric chorismate mutase is locked in the activated state by a single amino acid substitution

Cellulolytic Enzymes Production via Solid-State Fermentation: Effect of Pretreatment Methods on Physicochemical Characteristics of Substrate.

Chorismate mutase, a branch-point enzyme in the aromatic amino acid pathway of Saccharomyces cerevisiae, and also a mutant chorismate mutase with a single amino acid substitution in the C-terminal part of the protein have been purified approximately 20-fold and 64-fold from overproducing strains, respectively. The wild-type enzyme is activated by tryptophan and subject to feedback inhibition by tyrosine, whereas the mutant enzyme does not respond to activation by tryptophan nor inhibition by tyrosine. Both enzymes are dimers consisting of two identical subunits of M_r 30,000, each one capable of binding one substrate and one activator molecule. Each subunit of the wild-type enzyme also binds one inhibitor molecule, whereas the mutant enzyme lost this ability. The enzyme reaction was observed by "1H NMR and shows a direct and irreversible ...

53

Homology analyses of the protein sequences of fatty acid synthases from chicken liver, rat mammary gland, and yeast

We investigated the effect of pretreatment on the physicochemical characteristics-crystallinity, bed porosity, and volumetric specific surface of soybean hulls and production of cellulolytic enzymes in solid-state fermentation of Trichoderma reesei and Aspergillus oryzae cultures. Mild acid and alkali and steam pretreatments significantly increased crystallinity and bed porosity without significant change inholocellulosic composition of substrate. Crystalline and porous steam-pretreated soybean hulls inoculated with T. reesei culture had 4 filter paper units (FPU)/g-ds, 0.6?IU/g-ds ?-glucosidase, and 45?IU/g-ds endocellulase, whereas untreated hulls had 0.75?FPU/g-ds, 0.06?IU/g-ds ?-glucosidase, and 7.29?IU/g-ds endocellulase enzyme activities. In A. oryzae steam-pretreated soybean hulls had 47.10?IU/g-ds endocellulase compared to 30.82?IU/g-ds in untreated soybean hulls. Generalized linear statistical model fitted to ...

2011-06-15

54

Topoisomerases of kinetoplastid parasites: why so fascinating?

Homology analyses of the protein sequences of chicken liver and rat mammary gland fatty acid synthases were carried out. The amino acid sequences of the chicken and rat enzymes are 67% identical. If conservative substitutions are allowed, 78% of the amino acids are matched. A region of low homologies exists between the functional domains, in particular around amino acid residues 1059-1264 of the chicken enzyme. Homologies between the active sites of chicken and rat and of chicken and yeast enzymes have been analyzed by an alignment method. A high degree of homology exists between the active sites of the chicken and rat enzymes. However, the chicken and yeast enzymes show a lower degree of homology. The DADPH-binding dinucleotide folds of the {beta}-ketoacyl reductase and the enoyl reductase sites were identified by comparison with a known consensus sequence for the DADP- and ...

1989-11-01

55

British Library Electronic Table of Contents (United Kingdom)

Summary DNA topoisomerases are the key enzymes involved in carrying out high precision DNA transactions inside the cells. However, they are detrimental to the cell when a wide variety of topoisomerase-targeted drugs generate cytotoxic lesions by trapping the enzymes in covalent complexes on the DNA. The discovery of unusual heterodimeric topoisomerase I in kinetoplastid family added a new twist in topoisomerase research related to evolution, functional conservation and their preferential sensitivity to Camptothecin. On the other hand, structural and mechanistic studies on kinetoplastid topoisomerase II delineate some distinguishing features that differentiate the parasitic enzyme from its prokaryotic and eukaryotic counterparts. This review summarizes the recent advances in research in kin...

2006-01-01

56

The vacuolar processing enzyme (VPE) mutation suppresses an HR-like cell death induced by the double knockout mutant of vacuolar Ca2+-ATPases in Arabidopsis

The role of fatty acid desaturases in epidermal metabolism

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2011-06-01

57

Telomeric recombination induced by dysfunctional telomeres

The lipid composition of skin is important to a variety of functions served by this organ. Therefore, skin expresses multiple enzymes that synthesize and metabolize lipids. An important class of lipid...Full Text Available

2011-04-01

58

Solid-state fermentation of lignocellulotic materials for the production of enzymes by the white-rot fungus Trametes hirsuta in a modular bioreactor

Telomere maintenance is essential for cellular immortality, and most cancer cells maintain their telomeres through the enzyme telomerase. Telomeres and telomerase represent promising anticancer...Full Text Available

2011-01-15

59

British Library Electronic Table of Contents (United Kingdom)

Abstract Trametes hirsuta, a filamentous basidiomycete, was successfully cultivated in solid-state fermentation (SSF) on a mixture of pine wood chips and orange peel in a novel bioreactor that allows mixing of the inoculated solid material during the fermentation. Copper sulfate or xylidine (2,5-dimethylaniline) were added to enhance the production of enzymes, especially laccases. For comparison, Trametes hirsuta was also cultivated in submerged conditions. The effect of additives in SSF was low, whereas the choice of the solid material and the rotation of the reactor vessel showed a significant influence on the enzyme production. The space-time yields for the fermentations were calculated and showed that SSF on low-cost substrates can effectively produce extracellular enzymes at a suffici...

2011-01-01

60

Pressure Stabilization of Proteins from Extreme Thermophiles

One-electron reduction of chloroperoxidase by radiolytically generated electrons

We describe the stabilization by pressure of enzymes, including a hydrogenase from Methanococcus jannaschii, an extremely thermophilic deep-sea methanogen. This is the first published...Full Text Available

1994-03-01

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One-electron reduction of chloroperoxidase by radiolytically generated electrons

Upon irradiation of aqueous ethylene glycol/water solutions of native chloroperoxidase (CPO) with {sup 60}Co-gamma rays at 77K one observes the one-electron reduction of the enzyme active site by radiolytically generated thermolyzed electrons. In the present study the first absorption spectrum of a low-spin ferrous form of CPO is reported which has peaks at 438, 532 and 563 nm, similar to those observed previously for cytochrome P-450. All previously described ferrous forms of CPO are high spin. In order to observe the final results of the CPO reaction with electrons, the spectral changes of native enzyme after room temperature-gamma-irradiation have also been investigated. Evidence of changes is also presented probably connected with disruption of the tertiary structure of enzyme, correlated with decrease of enzyme activity.

1990-05-16

62

Kinetics of inhibition of green crab (Scylla serrata) alkaline phosphatase by sodium (2,2'-bipyridine) oxodiperoxovanadate.

Upon irradiation of aqueous ethylene glycol/water solutions of native chloroperoxidase (CPO) with "6"0Co-gamma rays at 77K one observes the one-electron reduction of the enzyme active site by radiolytically generated thermolyzed electrons. In the present study the first absorption spectrum of a low-spin ferrous form of CPO is reported which has peaks at 438, 532 and 563 nm, similar to those observed previously for cytochrome P-450. All previously described ferrous forms of CPO are high spin. In order to observe the final results of the CPO reaction with electrons, the spectral changes of native enzyme after room temperature-gamma-irradiation have also been investigated. Evidence of changes is also presented probably connected with disruption of the tertiary structure of enzyme, correlated with decrease of enzyme activity.

1990-05-01

63

Hormone-Sensitive Lipase Knockouts

Green crab (Scylla serrata) alkaline phosphatase (EC 3.1.3.1) is a metalloenzyme, which catalyzes the nonspecific hydrolysis of phosphate monoesters. The kinetics of inhibition of the enzyme by sodium (2, 2'-bipyridine) oxodiperoxovanadate, pV(bipy), has been studied. The time course of the hydrolysis of p-nitrophenyl-phosphate catalyzed by the enzyme in the presence of different pV(bipy) concentrations showed that at each pV(bipy) concentration, the rate decreased with increasing time until a straight line was approached, the straight line slopes are the same for all concentrations. The results suggest that the inhibition of the enzyme by pV(bipy) is a slow, reversible reaction with fractional remaining activity. The microscopic rate constants are determined for the reaction of inhibitor with the enzyme. PMID:10691182

1999-10-01

64

Evaluating concentration estimation errors in ELISA microarray experiments

All treatments for obesity, including dietary restriction of carbohydrates, have a goal of reducing the storage of fat in adipocytes. The chief enzyme responsible for the mobilization of FFA from adipose...Full Text Available

65

Cardiac Enzymes, Renal Failure and Renal Transplantation

BackgroundEnzyme-linked immunosorbent assay (ELISA) is a standard immunoassay to estimate a protein's concentration in a sample. Deploying ELISA in a microarray format permits simultaneous...Full Text Available

66

Analysis of Mammalian Carboxylesterase Inhibition by Trifluoromethylketone-Containing Compounds

Diagnostic accuracy of the currently available serum markers of cardiac injury, such as myoglobin, creatine kinase and its myocardial isoform, are altered in patients with renal failure. It is shown...Full Text Available

2006-03-01

67

The Acidic Domain of Hepatitis C Virus NS4A Contributes to RNA Replication and Virus Particle Assembly?

Carboxylesterases (CE) are ubiquitous enzymes that hydrolyze numerous ester-containing xenobiotics, including complex molecules, such as the anticancer drugs irinotecan (CPT-11) and capecitabine...Full Text Available

2007-03-01

68

Synthesis of non-steroidal anti-inflammatory drug analogues for selective studies on the COX-II enzyme

Hepatitis C virus NS3-4A is a membrane-bound enzyme complex that exhibits serine protease, RNA helicase, and RNA-stimulated ATPase activities. This enzyme complex is essential for viral genome replication...Full Text Available

2011-02-01

69

Signal transduction pathway controlling synthesis of a class of degradative enzymes in Bacillus subtilis: expression of the regulatory genes and analysis of mutations in degS and degU.

Synthesis of the azido substituted non-steroidal anti-inflammatory drug 2-(2,6-dichloroanilino)phenylacetic acid and isotope labeling of this compound have been performed and are described. Initial evaluation of the binding ability and photoreactivity indicates that this compound has potential for photoaffinity labeling as well as enzyme selectivity studies. (author).

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Sensitive enzyme immunoassay for detecting immunoglobulin M antibodies to Sindbis virus and further evidence that Pogosta disease is caused by a western equine encephalitis complex virus.

The rates of synthesis of a class of both secreted and intracellular degradative enzymes in Bacillus subtilis are controlled by a signal transduction pathway defined by at least four regulatory genes:...Full Text Available

1990-02-01

71

Radial distribution of dose and cross-sections for the inactivation of dry enzymes and viruses

An antibody capture enzyme immunoassay (EIA) was adapted for the detection of immunoglobulin M (IgM) antibody to Sindbis (SIN) virus. Sera from humans with a febrile illness characterized by rash and...Full Text Available

1985-10-01

72

Purification by Immunoadsorption and Immunochemical Properties of NADP-Dependent Malic Enzymes from Leaves of C₃, C₄, and Crassulacean Acid Metabolism Plants

A new semi-empirical algorithm for the radial distribution of dose is compared with available data. The algorithm is used to calculate the inactivation cross section for dry enzymes and viruses using an extended target model of a 1-hit detector. Agreement with data is at about the 15% level, approximating the precision of the data itself. (author).

1985-05-01

73

Physiological effects of manipulating the level of insulin-degrading enzyme in insulin-producing cells of Drosophila

NADP:malic enzyme from corn (Zea mays L.) leaves was purified by conventional techniques to apparent homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis....Full Text Available

1990-03-01

74

Evolution of a bifunctional enzyme: 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase.

Insulin-degrading enzyme (IDE) degrades insulin and other peptides, including the Aβ peptide of Alzheimer’s disease. However, the mechanism by which IDE acts on its substrates...Full Text Available

2011-01-01

75

Enzymic Synthesis of Indole-3-Acetyl-1-O-?-d-Glucose ¹

The bifunctional rat liver enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (ATP:D-fructose-6-phosphate 2-phosphotransferase/D-fructose-2,6-bisphosphate 2-phosphohydrolase, EC 2.7.1.105/EC...Full Text Available

1989-12-01

76

Determination by enzyme-linked immunosorbent assay of immunoglobulin G (IgG), IgM, and IgA to Brucella melitensis major outer membrane proteins and whole-cell heat-killed antigens in sera of patients with brucellosis.

The first enzyme-catalyzed reaction leading from indole-3-acetic acid (IAA) to the myo-inositol esters of IAA is the synthesis of indole-3-acetyl-1-O-β-d-glucose...Full Text Available

1988-12-01

77

DNA-new combination: a practical application in the sugar industry

An enzyme-linked immunosorbent assay was used to compare Brucella melitensis major outer membrane proteins (MOMP) and whole-cell heat-killed antigens (HK) in measuring antibrucella immunoglobulin G...Full Text Available

1989-08-01

78

DNA adducts in peripheral blood lymphocytes from aluminum production plant workers determined by 32P-postlabeling and enzyme-linked immunosorbent assay.

One of the first technical applications of gene technology is reported here. The DNA of E. coli cells was rearranged such that these cells produce large quantities of the enzyme ..cap alpha..-galactosidase. This enzyme is used to split the trisaccharide, raffinose, which is formed during the production of sugar from sugar beet. By this means sugar yields can be increased and the energy required for sugar crystallisation reduced.

1983-04-01

79

Comparison of indirect immunofluorescent-antibody assay, enzyme-linked immunosorbent assay, and Western immunoblot for the diagnosis of Lyme disease in dogs.

32P-Postlabeling analysis and enzyme-linked immunosorbent assay (ELISA) have been used to detect DNA adducts in peripheral blood lymphocytes from primary aluminum production plant workers who were exposed...Full Text Available

1993-03-01

80

Changes in the profile structure of the sarcoplasmic reticulum membrane induced by phosphorylation of the Ca2+ ATPase enzyme in the presence of terbium: a time-resolved x-ray diffraction study.

Enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescent-antibody assay (IFA), and Western immunoblot were used to test serum samples from 128 dogs for the presence of antibody to Borrelia...Full Text Available

1990-01-01

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81

Quantum Transition State Theory for proton transfer reactions in enzymes

The design of the time-resolved x-ray diffraction experiments reported in this and an accompanying paper was based on direct measurements of enzyme phosphorylation using [gamma-32P]ATP that were employed...Full Text Available

1994-05-01

82

CERN Document Server

We consider the role of quantum effects in the transfer of hyrogen-like species in enzyme-catalysed reactions. This study is stimulated by claims that the observed magnitude and temperature dependence of kinetic isotope effects imply that quantum tunneling below the energy barrier associated with the transition state significantly enhances the reaction rate in many enzymes. We use a path integral approach which provides a general framework to understand tunneling in a quantum system which interacts with an environment at non-zero temperature. Here the quantum system is the active site of the enzyme and the environment is the surrounding protein and water. Tunneling well below the barrier only occurs for temperatures less than a temperature $T_0$ which is determined by the curvature of potential energy surface near the top of the barrier. We argue that for most enzymes this temperature is less than room ...

2009-01-01

83

GENERIC ENZYME TECHNOLOGY FOR IMPROVED PAPER RECYCLING

Environmental Research Database

Objectives- Investigate the potential for the use of enzymes to bring about improvements in the recycling of recovered waste paper sources~%~~%~- Optimise enzyme formulations and process conditions to maximise benefits. - Produce recovered fibre at lower energy and chemical costs than is currently achievable using 'classical' de-inking techniques.~%~~%~- Improve recycled fibre quality and~%~~%~- All of the above will allow a greater use of recovered paper resources, which would otherwise be diposed of by [continued...]DescriptionThe project proposes to carry out an initial literature review to update the consortium on the currently available publications relating to the use of enzymes in de-inking.. This will be followed by an intensive laboratory study in an upgraded facility which will allow enzyme formulations and mechanisms to be studied. This ...

2003-01-05

84

Sucrose synthesizing enzymes and /sup 14/C-assimilation in the chlorophyllous layer of developing grapefruit. [Citrus paradisi Macf

Optimized assay and storage conditions for enzyme activity profiling of ectomycorrhizae

Fixation of /sup 14/CO/sub 2/ and activities of sucrose-synthesizing enzymes, sucrose phosphate synthetase (SPS) and sucrose synthase (SS), were assayed in tissues of developing fruit and source leaves from Citrus paradisi Macf. SPS activity of both the outer, chlorophyllous layer of the fruit (flavedo) and source leaves was 10-fold greater than that of the inner, largely non-chlorophyllous layer of the fruit peel (albedo). In contrast, SS activity of the flavedo was 2-fold greater than that of the albedo and 10-fold greater than that of leaves. Fixation of /sup 14/C-photosynthates in isolated tissues (flavedo 2x > albedo) and their redistribution in intact fruit indicated that flavedo functions as both source and sink. Activities of sucrose-synthesizing enzymes were consistent with this dual function.

1986-04-01

85

British Library Electronic Table of Contents (United Kingdom)

The aim of a joint effort by different research teams was to provide an improved procedure for enzyme activity profiling of field-sampled ectomycorrhizae, including recommendations on the best conditions and maximum duration for storage of ectomycorrhizal samples. A more simplified and efficient protocol compared to formerly published procedures was achieved by using manufactured 96-filter plates in combination with a vacuum manifold and by optimizing incubation times. Major improvements were achieved by performing the series of eight enzyme assays with a single series of root samples instead of two series, reducing the time needed for sample preparation, minimizing error-prone steps such as pipetting and morphotyping, and facilitating subsequent DNA analyses due to the reduced sequencing ...

2011-01-01

86

Effect of age-at-weaning on digestive capacity of white seabream (Diplodus sargus)

British Library Electronic Table of Contents (United Kingdom)

White seabream (Diplodus sargus) is today recognized as a potential species for Mediterranean aquaculture. Still, scarce information exists on weaning practices in order to reduce the live food period. This study aims to evaluate the effect of an early weaning on fish larvae digestive enzymes activities. In order to accomplish this, larvae were weaned with an inert diet at 20days after hatching (DAH) (feeding regime W20) and a control group was weaned at 27DAH (feeding regime W27). Before weaning, the onset and development of the main digestive enzymes were studied. The pattern of variation of digestive enzymes activities were analyzed at 0, 2, 9, 13 and 20DAH, and from then on at 0, 1 and 3weeks after diet introduction; that is, at 20, 27 and 41days for feeding regime W20, and days 27, 34...

2010-01-01

87

Vitamin E Is Essential for Seed Longevity and for Preventing Lipid Peroxidation during Germination

Uracil DNA glycosylase uses DNA hopping and short-range sliding to trap extrahelical uracils

Tocopherols (vitamin E) are lipophilic antioxidants synthesized by all plants and are particularly abundant in seeds. Despite cloning of the complete suite of tocopherol biosynthetic enzymes and successful...Full Text Available

2004-06-01

88

Understanding Enzyme Activity Using Single Molecule Tracking (Poster)

The astonishingly efficient location and excision of damaged DNA bases by DNA repair glycosylases is an especially intriguing problem in biology. One example is the enzyme uracil DNA glycosylase (UNG),...Full Text Available

2008-08-05

89

Two tandemly linked identical genes code for the glycosomal glyceraldehyde-phosphate dehydrogenase in Trypanosoma brucei.

This poster describes single-molecule tracking and total internal reflection fluorescence microscopy. It discusses whether the carbohydrate-binding module (CBM) moves on cellulose, how the CBM binds to cellulose, and the mechanism of cellulosome assembly.

2009-06-01

90

Toxic Myopathy in a Dog Associated with the Presence of Monensin in Dry Food

Trypanosoma brucei contains two isoenzymes for glyceraldehyde-phosphate dehydrogenase (GAPDH); one enzyme resides in a microbody-like organelle, the glycosome, the other one is found in the cytosol....Full Text Available

1986-05-01

91

The endogenous hydrogen sulfide producing enzyme cystathionine-? synthase contributes to visceral hypersensitivity in a rat model of irritable bowel syndrome

This report describes a case of toxic myopathy in a two year old sheltie dog with clinical signs of profound weakness, myoglobinuria, and muscle enzyme elevations. The clinical signs were likely related...Full Text Available

1980-01-01

92

The effects of zinc deficiency on pancreatic carboxypeptidase activity and protein digestion and absorption in the rat

BackgroundThe pathogenesis of visceral hypersensitivity, a characteristic pathophysiological feature of irritable bowel syndrome (IBS), remains elusive. Recent studies suggest a...Full Text Available

93

The control of sulphate reduction in bacteria

1. Proteolytic enzyme activities were examined in the pancreas of zinc-deficient and control rats. 2. No change was detected in trypsin-plus-chymotrypsin activity. 3. Carboxypeptidase activity was...Full Text Available

1967-03-01

94

The antihypertensive efficacy and tolerability of a low dose combination of ramipril and felodipine ER in mild to moderate essential hypertension.

1. An enzyme from Escherichia coli 9723 that reduces adenosine 3′-phosphate 5′-sulphatophosphate to inorganic sulphite is described. Extracts of E. coli...Full Text Available

1965-07-01

95

The Pregnane X Receptor: From Bench to Bedside

1. The antihypertensive efficacy and tolerability of a low dose combination of the angiotensin converting enzyme inhibitor ramipril (2.5 mg) and the extended release formulation of the dihydropyridine...Full Text Available

1993-10-01

96

The Mitochondrial Isovaleryl-Coenzyme A Dehydrogenase of Arabidopsis Oxidizes Intermediates of Leucine and Valine Catabolism1

BackgroundThe pregnane X receptor (PXR; NR1I2), a member of the nuclear receptor superfamily, regulates the expression of metabolic enzymes and transporters involved...Full Text Available

2008-07-01

97

Targeted plasmid integration into the human genome by an engineered zinc-finger recombinase

We recently identified a cDNA encoding a putative isovaleryl-coenzyme A (CoA) dehydrogenase in Arabidopsis (AtIVD). In animals, this homotetrameric enzyme is located in mitochondria and catalyzes the...Full Text Available

2001-06-01

98

Taking the plunge: integrating structural, enzymatic and computational insights into a unified model for membrane-immersed rhomboid proteolysis

The development of new methods for gene addition to mammalian genomes is necessary to overcome the limitations of conventional genetic engineering strategies. Although a variety of DNA-modifying enzymes...Full Text Available

2011-09-01

99

Subunits of the Drosophila CCR4-NOT complex and their roles in mRNA deadenylation

SYNOPSISRhomboid proteases are a fascinating class of enzymes that combine a serine protease active site within the core of an integral membrane protein. Despite having key roles...Full Text Available

100

Some Enzymic Activities in the Germinating Oil Palm (Elaeis guineensis) Seedling 1

The CCR4-NOT complex is the main enzyme catalyzing the deadenylation of mRNA. We have investigated the composition of this complex in Drosophila melanogaster by immunoprecipitation...Full Text Available

2010-07-01

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101

Soluble Epoxide Hydrolase Inhibition: Targeting Multiple Mechanisms of Ischemic Brain Injury with a Single Agent

In germinating oil palm (Elaeis guineensis var D × P) seedling, an active lipase was present in the shoot but absent from both the kernel and the haustorium. It has an optimum...Full Text Available

1983-12-01

102

Single-Step Production of a Recyclable Nanobiocatalyst for Organophosphate Pesticides Biodegradation Using Functionalized Bacterial Magnetosomes

SummarySoluble epoxide hydrolase (sEH) is a key enzyme in the metabolic conversion and degradation of P450 eicosanoids called epoxyeicosatrienoic acids (EETs). Genetic variations...Full Text Available

2009-03-01

103

Reversible Inactivation of Nitrate Reductase by NADH and the Occurrence of Partially Inactive Enzyme in the Wheat Leaf 1

Enzymes are versatile catalysts in laboratories and on an industrial scale; improving their immobilization would be beneficial to broadening their applicability and ensuring their (re)use. Lipid-coated...Full Text Available

104

Regulation of lactose catabolism in Streptococcus mutans: purification and regulatory properties of phospho-beta-galactosidase.

Nitrate reductase from wheat (Triticum aestivum L. cv Bindawarra) leaves is inactivated by pretreatment with NADH, in the absence of nitrate, a 50% loss of activity occurring in 30...Full Text Available

1983-03-01

105

Recent changes in the epidemiology and management of extended-spectrum ?-lactamase-producing Enterobacteriaceae

Phospho-beta-galactosidase (P-beta-gal), the enzyme which catalyzes the first step in the metabolism of intracellular lactose phosphate, occurred at high specific activity in the cytoplasm in 12 of...Full Text Available

1979-01-01

106

Rare Homologous Gene Targeting in Histoplasma capsulatum: Disruption of the URA5_Hc Gene by Allelic Replacement

Since 2000, Escherichia coli producing CTX-M enzymes (especially CTX-M-15) have emerged worldwide as important causes of community-onset urinary tract and blood stream infections due...Full Text Available

107

Purification and properties of an endo-1,4-beta-glucanase from Clostridium josui.

URA5 genes encode orotidine-5′-monophosphate pyrophosphorylase (OMPpase), an enzyme involved in pyrimidine biosynthesis. We cloned the Histoplasma capsulatum URA5...Full Text Available

1998-10-01

108

Production, Purification, and Characterization of ?-Galactosidase from Monascus pilosus

An enzyme active against carboxymethyl cellulose (CMC) was purified from the stationary-phase-culture supernatant of Clostridium josui grown in a medium containing ball-milled cellulose. The purification...Full Text Available

1989-07-01

109

Preliminary X-ray crystallographic studies of Bacillus subtilis SpeA protein

A Monascus pilosus strain was selected for production of intracellular α-galactosidase. Optimum conditions for mycelial growth and enzyme induction were determined. Galactose...Full Text Available

1986-11-01

110

Polyamine Oxidase from Water Hyacinth

The speA gene in Bacillus subtilis encodes arginine decarboxylase, which catalyzes the conversion of arginine to agmatine. Arginine decarboxylase is an important enzyme...Full Text Available

111

Plant and algal interference in bacterial beta-D-galactosidase and beta-D-glucuronidase assays.

Polyamine oxidase was purified to homogeneity from leaves of water hyacinth by the criterion of sodium dodecyl sulfate gel electrophoresis (SDS disc PAGE). The enzyme showed a high specificity for spermidine...Full Text Available

1987-12-01

112

Physiological calcium concentrations regulate calmodulin binding and catalysis of adenylyl cyclase exotoxins

Several commonly occurring freshwater and marine plants and algae were screened for beta-D-galactosidase and beta-D-glucuronidase activities by using a 60-min enzyme assay based on the hydrolysis by...Full Text Available

1994-11-01

113

Permeability of frog mesenteric capillaries after partial pronase digestion of the endothelial glycocalyx.

Edema factor (EF) and CyaA are calmodulin (CaM)-activated adenylyl cyclase exotoxins involved in the pathogenesis of anthrax and whooping cough, respectively. Using spectroscopic, enzyme kinetic and...Full Text Available

2002-12-16

114

Peripartal changes in serum alkaline phosphatase activity and lactate dehydrogenase activity in dairy cows.

1. The proteolytic enzyme pronase, which degrades the endothelial cell glycocalyx, was perfused through single capillaries of frog mesentery. Hydraulic conductivity (Lp) of each vessel was determined...Full Text Available

1990-09-01

115

Pathogenic Role of NKT and NK Cells in Acetaminophen-Induced Liver Injury is Dependent on the Presence of DMSO

Peripartal serum alkaline phosphatase activity and lactate dehydrogenase activity were measured in 30 dairy cows in order to examine the association between retained fetal membranes and enzyme activity....Full Text Available

1987-10-01

116

On the General Mechanism of Selective Induction of Cytochrome P450 Enzymes by Chemicals: Some Theoretical Considerations

Dimethyl sulfoxide (DMSO) is commonly used in biological studies to dissolve drugs and enzyme inhibitors with low solubility. While DMSO is generally thought of as being relatively inert, it...Full Text Available

2008-09-01

117

Neutral endopeptidase inhibits prostate cancer cell migration by blocking focal adhesion kinase signaling

Importance of the fieldThe cytochrome P450 (CYP) isoforms that are selectively induced following exposure to structurally-diverse chemicals often are the ones capable...Full Text Available

2010-04-01

118

NasFED Proteins Mediate Assimilatory Nitrate and Nitrite Transport in Klebsiella oxytoca (pneumoniae) M5al

Neutral endopeptidase 24.11 (NEP, CD10) is a cell-surface enzyme expressed by prostatic epithelial cells that cleaves and inactivates neuropeptides implicated in the growth of androgen-independent prostate...Full Text Available

2000-12-01

119

N-acetyltransferase 2 slow acetylator genotype associated with adverse effects of sulfasalazine in the treatment of inflammatory bowel disease

Klebsiella oxytoca can use nitrate and nitrite as sole nitrogen sources. The enzymes required for nitrate and nitrite assimilation are encoded by the nasFEDCBA operon....Full Text Available

1998-03-01

120

Molecular mechanisms of genetic adaptation to xenobiotic compounds.

AIM:N-acetyltransferase 2 (NAT2) is an important enzyme catalyzing N-acetylation of sulfasalazine (SASP). The aim of the present study was to investigate...Full Text Available

2007-03-01

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121

Molecular cloning and nucleotide sequence of the beta-lytic protease gene from Achromobacter lyticus.

Microorganisms in the environment can often adapt to use xenobiotic chemicals as novel growth and energy substrates. Specialized enzyme systems and metabolic pathways for the degradation of man-made...Full Text Available

1992-12-01

122

Molecular basis of the inhibition of human aromatase (estrogen synthetase) by flavone and isoflavone phytoestrogens: A site-directed mutagenesis study.

Two bacteriolytic enzymes secreted by Achromobacter lyticus M497-1 were purified and identified as being very similar (considering their amino acid composition and N-terminal sequence) to alpha- and...Full Text Available

1990-11-01

123

Molecular and Kinetic Comparison of the Novel Extended-Spectrum ?-Lactamases CTX-M-25 and CTX-M-26

Flavone and isoflavone phytoestrogens are plant chemicals and are known to be competitive inhibitors of cytochrome P450 aromatase with respect to the androgen substrate. Aromatase is the enzyme that...Full Text Available

1998-02-01

124

Mitochondrial DNA Damage and Animal Longevity: Insights from Comparative Studies

CTX-M-25 is a novel extended-spectrum β-lactamase isolated from a single Canadian Escherichia coli isolate. Susceptibility testing demonstrated that this enzyme confers resistance...Full Text Available

2004-12-01

125

Chemical reactions in living cells are under strict enzyme control and conform to a tightly regulated metabolic program. However, uncontrolled and potentially deleterious endogenous reactions occur,...Full Text Available

126

Metabolism of Cytokinin

Lead and ?-Aminolevulinic Acid Dehydratase Polymorphism: Where Does It Lead? A Meta-Analysis

As part of the study of cytokinin metabolic pathways, an enzyme, adenosine phosphorylase (EC 2.4.2.-), which catalyzed the ribosylation of N⁶-(Δ²-isopentenyl)adenine,...Full Text Available

1978-12-01

127

Kinetics of complexing activation by the magnesium ion on green crab (Scylla serrata) alkaline phosphatase.

BackgroundLead poisoning affects many organs in the body. Lead inhibits δ-aminolevulinic acid dehydratase (ALAD), an enzyme with two co-dominantly expressed alleles, ALAD1...Full Text Available

2007-01-01

128

Isozymes of the Glycolytic Enzymes in Endosperm from Developing Castor Oil Seeds ¹

As with mammalian enzymes, green crab (Scylla serrata) alkaline phosphatase can be activated by Mg2+ through a time-dependent course. The activation is mainly a Vmax effect. Tsou's method was used to study the kinetic course of activation. The results show that the enzyme was activated by a complexing scheme that had not been previously identified: the enzyme first reversibly and quickly binds Mg2+ and then undergoes a slow reversible course to activation, with a relatively high activation energy (78 +/- 4 kJ/mol) and a slow conformational change. The activation reaction is a single molecule reaction, and the apparent activation rate constant is independent of Mg2+ concentration if the concentration is sufficiently high. The microscopic rate constants of activation and the association constant were determined from the measurements. The proposed scheme may also be applied to the Mg2+ activation mechanism for mammalian ...

2001-01-01

129

Involvement of chlA, E, M, and N loci in Escherichia coli molybdopterin biosynthesis.

Ion filtration chromatography on diethylaminoethyl-Sephadex A-25 has been used to separate two isozymes each of triose phosphate isomerase, glyceraldehyde 3-phosphate dehydrogenase, glycerate 3-phosphate...Full Text Available

1982-04-01

130

Indoleamine 2,3-dioxygenase-expressing leukemic dendritic cells impair a leukemia-specific immune response by inducing potent T regulatory cells

All molybdenum enzymes except nitrogenase contain a common molybdenum cofactor, whose organic moiety is a novel pterin called molybdopterin (MPT). To assist in elucidating the biosynthetic pathway of...Full Text Available

1987-01-01

131

Increased Expression and Activity of Matrix Metalloproteinases Characterize Embolic Cardiac Myxomas

BackgroundThe immunoregulatory enzyme indoleamine 2,3-dioxygenase, which catalyzes the conversion of tryptophan into kynurenine, is expressed in a significant subset of patients...Full Text Available

2010-12-01

132

Incidence and Determinants of Birth Defects and Enzyme Deficiencies among Live Births in Oman

Tumor embolism occurs in 30 to 50% of all cases of cardiac myxoma, but the causes are still uncertain. Matrix metalloproteinases (MMPs) are proteolytic enzymes that degrade the extracellular matrix...Full Text Available

2005-06-01

133

Identification of PITX1 as a TERT Suppressor Gene Located on Human Chromosome 5 ^?

ObjectivesIn 2003, the Omani Ministry of Health Child Health Care Program initiated a national Birth Defects (BD) Register. This paper reviews the magnitude and risk factors of birth...Full Text Available

2010-04-01

134

Hysteretic Behavior of Proprotein Convertase 1/3 (PC1/3)

Telomerase, a ribonucleoprotein enzyme that maintains telomere length, is crucial for cellular immortalization and cancer progression. Telomerase activity is attributed primarily to the expression of...Full Text Available

2011-04-01

135

Host plant shifts affect a major defense enzyme in Chrysomela lapponica

The proprotein convertases (PCs) are calcium-dependent proteases responsible for processing precursor proteins into their active forms in eukariotes. The PC1/3 is a pivotal enzyme of this family that...Full Text Available

136

Gynura procumbens Merr. decreases blood pressure in rats by vasodilatation via inhibition of calcium channels

Chrysomelid leaf beetles use chemical defenses to overcome predatory attack and microbial infestation. Larvae of Chrysomela lapponica that feed on willow sequester plant-derived salicin...Full Text Available

2011-03-22

137

Glutamate 2,3-aminomutase: a new member of the radical SAM superfamily of enzymes

INTRODUCTION:Gynura procumbens has been shown to decrease blood pressure via inhibition of the angiotensin‐converting enzyme. However, other mechanisms that...Full Text Available

2011-01-01

138

Genetics of the mammalian phenylalanine hydroxylase system. Studies of human liver phenylalanine hydroxylase subunit structure and of mutations in phenylketonuria.

SUMMARYA gene eam in Clostridium difficile encodes a protein that is homologous to lysine 2,3-aminomutase (LAM) in many other species but does...Full Text Available

2007-02-01

139

Gene-enzyme telationships in somatic cells and their organismal derivatives in higher plants. Progress report

Phenylalanine hydroxylase was purified from crude extracts of human livers which show enzyme activity by usine two different methods: (a) affinity chromatography and (b) immunoprecipitation with an...Full Text Available

1979-08-01

140

Functional and Structural Analysis of a Key Region of the Cell Wall Inhibitor Moenomycin

Progress is reported in the following subject areas: (1) chemistry of the arogenate molecule; (2) plant enzymology at the organismal level; (3) isolation of regulatory mutants in tobacco; and (4) stability of the haploid state in Nicotiana sylvestris.

1980-04-21

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141

Formate Dehydrogenase of Clostridium thermoaceticum: Incorporation of Selenium-75, and the Effects of Selenite, Molybdate, and Tungstate on the Enzyme

Moenomycin A (MmA) belongs to a family of natural products that inhibit peptidoglycan biosynthesis by binding to the peptidoglycan glycosyltransferases (PGTs), the enzymes that make the glycan...Full Text Available

2010-07-16

142

Finasteride-Its Impact on Sexual Function and Prostate Cancer

The formation of the nicotinamide adenine dinucleotide phosphate-dependent formate dehydrogenase in Clostridium thermoaceticum is stimulated by the presence of molybdate and selenite...Full Text Available

1973-11-01

143

Fermentability of Corn Syrups with Different Dextrose Equivalents Added to Various Grape Juices¹

Finasteride, a specific and competitive inhibitor of 5α-reductase enzyme Type 2, inhibits the conversion of testosterone to dihydrotestosterone (DHT). In adults, DHT acts as primary androgen...Full Text Available

2009-01-01

144

Expression and prognostic significance of cox-2 and p-53 in hodgkin lymphomas: a retrospective study

It was found that neither the enzymes of the grapes nor those of wine yeast Saccharomyces ellipsoideus strain 223 attacked the higher polysaccharides present in corn syrups. The alcohol...Full Text Available

1967-03-01

145

Examination of the Rotazyme II enzyme immunoassay for the diagnosis of rotavirus gastroenteritis.

BackgroundCyclooxygenase (cox) is the rate-limiting enzyme, which catalyzes the conversion of arachidonic acid into prostaglandins and contributes to the inflammatory process. Cyclooxygenase-2...Full Text Available

146

Evolutionary history of a specialized P450 propane monooxygenase

Rotazyme II, which is a shorter version of Rotazyme (less than 3 h), was compared with electron microscopy and Rotazyme for sensitivity and specificity on 229 human stool specimens. Compared with electron...Full Text Available

1985-09-01

147

Evolutionary Trajectories of Beta-Lactamase CTX-M-1 Cluster Enzymes: Predicting Antibiotic Resistance

SummaryThe evolutionary pressures that shaped the specificity and catalytic efficiency of enzymes can only be speculated. While directed evolution experiments show that new functions...Full Text Available

2008-11-28

148

Evaluation of protector potential of extracts from Cymbopogon citratus, Maytenus ilicifolia, Baccharis genistelloides against the effects of stannous chloride: role of exonuclease III enzyme

Extended-spectrum beta-lactamases (ESBL) constitute a key antibiotic-resistance mechanism affecting Gram-negative bacteria, and also an excellent model for studying evolution in real time. A shift in...Full Text Available

2010-01-01

149

Enzymic Capacities of Purified Cauliflower Bud Plastids for Lipid Synthesis and Carbohydrate Metabolism ¹

Portuguese 2003 [1 p.] Brazil Soares, SF Melo, SF Costa, RF Silva, CR

2003-11-20

150

Ellman's-reagent-mediated regeneration of trypanothione in situ: substrate-economical microplate and time-dependent inhibition assays for trypanothione reductase.

Isolated cauliflower (Brassica oleracea) bud plastids, purified by isopycnic centrifugation in density gradients of Percoll, were found to be highly intact, to be practically devoid...Full Text Available

1985-10-01

151

Effects of PKA phosphorylation on the conformation of the Na,K-ATPase regulatory protein FXYD1

Trypanothione reductase (TryR) is a key enzyme involved in the oxidative stress management of the Trypanosoma and Leishmania parasites, which helps to maintain an intracellular reducing environment...Full Text Available

2003-02-01

152

Effects of Leupeptin on Proteinase and Germination of Castor Beans ¹

FXYD1 (phospholemman) is a member of an evolutionarily conserved family of membrane proteins that regulate the function of the Na,K-ATPase enzyme complex in specific tissues and specific physiological...Full Text Available

2009-11-01

153

Effects of Kombucha on oxidative stress induced nephrotoxicity in rats

Leupeptin, a tripeptide inhibitor of some proteinases, was shown previously to maintain the stability of several enzymes (isocitrate lyase, fumarase, and catalase) in crude extracts of castor bean endosperm....Full Text Available

1981-10-01

154

Effects of DGAT1 deficiency on energy and glucose metabolism are independent of adiponectin

BackgroundTrichloroethylene (TCE) may induce oxidative stress which generates free radicals and alters antioxidants or oxygen-free radical scavenging enzymes.MethodsTwenty...Full Text Available

155

Diurnal Variations of Mouse Plasma and Hepatic Bile Acid Concentrations as well as Expression of Biosynthetic Enzymes and Transporters

Mice lacking acyl-CoA:diacylglycerol acyltransferase 1 (DGAT1), an enzyme that catalyzes the terminal step in triacylglycerol synthesis, have enhanced insulin sensitivity and are protected from...Full Text Available

2006-08-01

156

Dissemination of CTX-M-Type Extended-Spectrum ?-Lactamase Genes to Unusual Hosts

BackgroundDiurnal fluctuation of bile acid (BA) concentrations in the enterohepatic system of mammals has been known for a long time. Recently, BAs have been recognized as signaling...Full Text Available

157

Developmental and metabolic regulation of the Drosophila melanogaster 3-hydroxy-3-methylglutaryl coenzyme A reductase.

A Citrobacter amalonaticus and a Morganella morganii producing the CTX-M-1 extended-spectrum β-lactamase (ESBL) were isolated from an area where this enzyme...Full Text Available

2005-08-01

158

Delineating Bacteriostatic and Bactericidal Targets in Mycobacteria Using IPTG Inducible Antisense Expression

The enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase in Drosophila melanogaster synthesizes mevalonate for the production of nonsterol isoprenoids, which are essential for growth and...Full Text Available

1988-07-01

159

Deglutathionylation of 2-Cys Peroxiredoxin Is Specifically Catalyzed by Sulfiredoxin*

In order to identify novel high value antibacterial targets it is desirable to delineate whether the inactivation of the target enzyme will lead to bacterial death or stasis. This knowledge is particularly...Full Text Available

160

Crystallization and preliminary X-ray analysis of human liver ?-enolase

Reversible protein glutathionylation plays a key role in cellular regulation and cell signaling and protects protein thiols from hyperoxidation. Sulfiredoxin (Srx), an enzyme that catalyzes the reduction...Full Text Available

2009-08-28

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161

Cottonseed Malate Synthase ¹

Enolase is a multifunctional enzyme that plays important roles in many biological and disease processes. α-Enolase from human liver (hENO1) was expressed as a soluble protein and purified by...Full Text Available

162

Coordination of PAD4 and HDAC2 in the regulation of p53 target gene expression

Malate synthase (EC 4.1.3.2), an enzyme unique to the glyoxylate cycle, was purified to homogeneity from cotyledons of 72-hours, darkgrown cotton (Gossypium hirsutum L.) seedlings....Full Text Available

1987-08-01

163

Complete sequence of the chicken glyceraldehyde-3-phosphate dehydrogenase gene.

Histone Arg methylation and Lys acetylation have been found to cooperatively regulate the expression of p53 target genes. Peptidylarginine deiminase 4 (PAD4) is an enzyme that citrullinates...Full Text Available

2010-05-27

164

Cloning and sequencing of a dehalogenase gene encoding an enzyme with hydrolase activity involved in the degradation of gamma-hexachlorocyclohexane in Pseudomonas paucimobilis.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an evolutionarily conserved glycolytic enzyme, is constitutively expressed in most cell types yet is induced to high levels during the development of...Full Text Available

1985-03-01

165

Clinical outcomes and management of mechanism-based inhibition of cytochrome P450 3A4

In Pseudomonas paucimobilis UT26, gamma-hexachlorocyclohexane (gamma-HCH) is converted by two steps of dehydrochlorination to a chemically unstable intermediate, 1,3,4,6-tetrachloro-1,4-cyclohexadiene...Full Text Available

1993-10-01

166

Chemoprevention of Nonmelanoma Skin Cancer With Celecoxib: A Randomized, Double-Blind, Placebo-Controlled Trial

Mechanism-based inhibition of cytochrome P450 (CYP) 3A4 is characterized by NADPH-, time-, and concentration-dependent enzyme inactivation, occurring when some drugs are converted by CYPs to reactive...Full Text Available

2005-03-01

167

Characterization of psychrotrophic microorganisms producing beta-galactosidase activities.

BackgroundPreclinical studies indicate that the enzyme cyclooxygenase 2 plays an important role in ultraviolet-induced skin cancers. We evaluated the efficacy and safety of celecoxib,...Full Text Available

2010-12-15

168

Changes in N-acetyl-B-D-glucosaminidase and B-glucuronidase activities in milk during bovine mastitis.

Investigations of psychrotrophic microorganisms have been limited even though the dominant environment of the Earth is cold and enzymes with high activities at low temperatures could have commercial...Full Text Available

1994-01-01

169

Cerebrospinal fluid zinc concentrations in febrile convulsions.

To determine the N-acetyl-B-D-glucosaminidase (NAGase) and B-glucuronidase (B-Gase) activities in mastitic milk, basic enzyme assay conditions, distribution of NAGase and B-Gase, comparison of their...Full Text Available

1987-01-01

170

Cell-density-dependent Changes in the Metabolism of Chloronema Cell Cultures

Zinc modulates the activity of glutamic acid decarboxylase, the rate limiting enzyme in the synthesis of gamma-aminobutyric acid (GABA), which is a major inhibitory neurotransmitter. Low cerebrospinal...Full Text Available

1995-10-01

171

Carotenoid Crystal Formation in Arabidopsis and Carrot Roots Caused by Increased Phytoene Synthase Protein Levels

In the growing chloronema cell suspension cultures of the moss Funaria hygrometrica Hedw., activities of several enzymes have been found to be cell-density-dependent. Cyclic nucleotide...Full Text Available

1979-07-01

172

Capacity of deoxycytidine to selectively antagonize cytotoxicity of 5-halogenated analogs of deoxycytidine without loss of antiherpetic activity.

BackgroundAs the first pathway-specific enzyme in carotenoid biosynthesis, phytoene synthase (PSY) is a prime regulatory target. This includes a number of biotechnological approaches...Full Text Available

173

CTX-M-Type Extended-Spectrum ?-Lactamases in Italy: Molecular Epidemiology of an Emerging Countrywide Problem

Enzyme kinetic studies from this laboratory (M. Dobersen and S. Greer, Biochemistry 17:920-928, 1978) suggested that deoxycytidine could antagonize the toxicity of 5-halogenated analogs of deoxycytidine...Full Text Available

1982-09-01

174

Betaxanthins as Substrates for Tyrosinase. An Approach to the Role of Tyrosinase in the Biosynthetic Pathway of Betalains1

A nationwide survey of extended-spectrum β-lactamase (ESBL) production among Enterobacteriaceae, carried out in 2003, showed that CTX-M-type enzymes have achieved a sizeable...Full Text Available

2006-08-01

175

Associations between dairy production indices and lipoarabinomannan enzyme-immunoassay results for paratuberculosis.

Tyrosinase or polyphenol oxidase (EC 1.14.18.1) is the key enzyme in melanin biosynthesis and in the enzymatic browning of fruits and vegetables. The role of tyrosinase in the secondary metabolism of...Full Text Available

2005-05-01

176

Association of COMT Val^108/158Met Genotype and Cigarette Smoking in Pregnant Women

Data from an epidemiological study in Ontario, involving 304 dairy herds, were used to identify associations between selected production indices and lipoarabinomannan antigen serological test results...Full Text Available

1991-10-01

177

Antioxidant enzyme activities are not broadly correlated with longevity in 14 vertebrate endotherm species

Introduction:Smoking behaviors, including heaviness of smoking and smoking cessation, are known to be under a degree of genetic influence. The enzyme catechol O-methyltransferase...Full Text Available

2011-02-01

178

Antibody avidity in swine lymphocyte antigen-defined miniature pigs.

The free radical theory of ageing posits that accrual of oxidative damage underlies the increased cellular, tissue and organ dysfunction and failure associated with advanced age. In support of this...Full Text Available

2010-06-01

179

Anti-beta-endorphin immunoglobulin G in humans.

Antibody avidity to hen egg white lysozyme (HEWL) was measured by thiocyanate ion elution enzyme-linked immunosorbent assay (ELISA) in swine lymphocyte antigen (SLA) defined miniature pigs. Serum antibody...Full Text Available

1992-10-01

180

Analysis of the Pharmacokinetic Interaction between Cephalexin and Quinapril by a Nonlinear Mixed-Effect Model

Human IgG specific for beta-endorphin was identified by enzyme-linked immunoabsorbent assay and isolated by affinity chromatography. From a sample of 27 subjects, three individuals with major depression...Full Text Available

1986-11-01

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181

Amastigote stage-specific monoclonal antibodies against Leishmania major.

Oligopeptidic drugs such as β-lactams and angiotensin-converting enzyme inhibitors share the same carriers in humans and animals, which results in possible pharmacokinetic interactions. To model...Full Text Available

1998-06-01

182

Adenosine 5?-triphosphate sulphurylase from Saccharomyces cerevisiae

Monoclonal antibodies were produced against gamma-irradiated amastigotes of Leishmania major. Five antibodies (T16 through T20) were selected which reacted in enzyme-linked immunoassays with the intracellular...Full Text Available

1989-12-01

183

Acetaldehyde and Ethanol Biosynthesis in Leaves of Plants ¹

1. ATP sulphurylase from Saccharomyces cerevisiae was purified 140-fold by using heat treatment, DEAE-cellulose chromatography and Sepharose 6B gel filtration. 2. The enzyme was stable...Full Text Available

1973-07-01

184

Absence of Ataxin-3 Leads to Enhanced Stress Response in C. elegans

Leaves of terrestrial plants are aerobic organs, and are not usually considered to possess the enzymes necessary for biosynthesis of ethanol, a product of anaerobic fermentation. We examined the ability...Full Text Available

1987-08-01

185

ADP-Glucose Pyrophosphorylase, a Regulatory Enzyme for Bacterial Glycogen Synthesis

Ataxin-3, the protein involved in Machado-Joseph disease, is able to bind ubiquitylated substrates and act as a deubiquitylating enzyme in vitro, and it has been involved in the modulation of protein...Full Text Available

186

A study in molecular contingency: glutamine phosphoribosylpyrophosphate amidotransferase is a promiscuous and evolvable phosphoribosylanthranilate isomerase

The accumulation of α-1,4-polyglucans is an important strategy to cope with transient starvation conditions in the environment. In bacteria and plants, the synthesis of glycogen and starch occurs...Full Text Available

2003-06-01

187

A novel enzyme-based acidizing system: Matrix acidizing and drilling fluid damage removal

SummaryThe prevalence of paralogous enzymes implies that novel catalytic functions can evolve on preexisting protein scaffolds. The weak secondary activities of proteins, which...Full Text Available

2008-03-21

188

A cytochrome P450 terpenoid hydroxylase linked to the suppression of insect juvenile hormone synthesis

A novel acidizing process is used to increase the permeability of carbonate rock cores in the laboratory and to remove drilling fluid damage from cores and wafers. Field results show the benefits of the technology as applied both to injector and producer wells.

1995-12-31

189

A Hyperactive NAD(P)H:Rubredoxin Oxidoreductase from the Hyperthermophilic Archaeon Pyrococcus furiosus

A cDNA encoding a cytochrome P450 enzyme was isolated from a cDNA library of the corpora allata (CA) from reproductively active Diploptera punctata cockroaches. This P450 from the endocrine...Full Text Available

1998-10-27

190

Identification of a nucleoside triphosphate binding site on calf thymus RNA polymerase II

NAD(P)H:rubredoxin oxidoreductase (NROR) has been purified from the hyperthermophilic archaeon Pyrococcus furiosus. The enzyme is exceedingly active in catalyzing the NADPH-dependent...Full Text Available

1999-09-01

191

Two and 8-azido photoaffinity probes. 2. Studies on the binding process of 2-5A synthetase by photosensitive ATP analogues

A nucleoside triphosphate binding site on calf thymus RNA polymerase II was identified by using photoaffinity analogues of adenosine 5'-triphosphate and guanosine 5'-triphosphate. Both radiolabeled 8-azidoadenosine 5'-triphosphate (8-N3ATP) and radiolabeled 8-azidoguanosine 5'-triphosphate (8-N3GTP) bound to a single polypeptide of this enzyme. This polypeptide has a molecular mass of 37 kilodaltons and an isoelectric point of 5.4. Ultraviolet (UV) irradiation was necessary for photolabeling to occur. In addition, no labeling occurred when the probe was prephotolyzed or when the enzyme was inactivated. Furthermore, photolabeling of the enzyme could be decreased by preincubation with natural substrates. To provide evidence that the radiolabeled polypeptide forms a part of the domain of the nucleoside triphosphate binding site, experiments were performed using unlabeled 8-N3ATP. Although ...

1986-01-14

192

Structures of trihydroxynaphthalene reductase-fungicide complexes: implications for structure-based design and catalysis

The photoaffinity probes [#gamma#-"3"2P]2-azidoATP (2-N_3ATP) and [#alpha#-"3"2P]8-azido-ATP (8-N_3ATP) were used to investigate the binding of ATP to highly purified 2-5A synthetase. 2-N_3APT and 8-N_3ATP are substrates for 2-5A synthetase. In this study the authors show that 2- and 8-N_3ATP are competitive inhibitors of the enzymatic conversion of ATP to 2-5A. Ultraviolet irradiation results in the photoinsertion of 2-N_3ATP and 8-N_3ATP into the enzyme. The covalent photoinsertion of [#alpha#-"3"2P]8-N_3ATP into the 2-5A synthetase is proportional to the inactivation of the enzyme as UV irradiation is increased. Photolabeling of 2-5A synthetase is saturated at 1.5 mM 2-N_3ATP and 2.0 mM 8-N_3ATP. Computer analysis of the curvilinear Scatchard plots of the 2-5A synthetase suggest the presence of high-affinity and low-affinity binding sites that may correspond to the acceptor and the 2'-adenylation sites of the enzyme. The ...

193

Mechanism of enzyme-catalyzed phospho group transfer

Trihydroxynaphthalene reductase catalyzes two intermediate steps in the fungal melanin biosynthetic pathway. The enzyme, a typical short-chain dehydrogenase, is the biochemical target of three commercial fungicides. The fungicides bind preferentially to the NADPH form of the enzyme. Three X-ray structures of the Magnaporthe grisea enzyme complexed with NADPH and two commercial and one experimental fungicide were determined at 1.7 {angstrom} (pyroquilon), 2.0 {angstrom} (2,3-dihydro-4-nitro-1H-inden-1-one, 1), and 2.1 {angstrom} (phthalide) resolutions. The chemically distinct inhibitors occupy similar space within the enzyme's active site. The three inhibitors share hydrogen bonds with the side chain hydroxyls of Ser-164 and Tyr-178 via a carbonyl oxygen (pyroquilon and 1) or via a carbonyl oxygen and a ring oxygen (phthalide). Active site residues occupy similar positions among the three ...

2010-03-08

194

Interplay Between Oncoproteins and Antioxidant Enzymes in Esophageal Carcinoma Treated Without and With Chemoradiotherapy: A Prospective Study

To understand more fully the mechanism of enzyme-catalyzed phospho group transfer, the stereochemical course at phosphorus of four enzymes has been determined. First, using adenosine (..gamma..-(S)-/sup 16/O, /sup 17/O, /sup 18/O)triphosphate as the substrate, the reaction catalyzed by creatine kinase has been found to proceed with overall inversion of configuration at phosphorus. Second, using adenosine (..beta..-(S)-/sup 16/O, /sup 17/O, /sup 18/O)diphosphate as the substrate, the reaction catalyzed by adenylate kinase has been found also to proceed with overall inversion. Third, the reaction catalyzed by phosphoenolpyruvate carboxylase has been studied using ((S/sub p/)-/sup 16/O, /sup 17/O)thiophospoenolpyruvate as the substrate in H/sub 2/ /sup 18/O. Fourth, using adenosine 5'-O-((..gamma..S/sub p/)-..beta gamma..-/sup 17/O,..gamma..-/sup 17/O,/sup 18/O)(3-thiotriphosphate) as the substrate, the reaction catalyzed by pyruvate ...

1986-01-01

195

Formation of an intramolecular cystine disulfide during the reaction of 8-azidoguanosine 5'-triphosphate with cytosolic phosphoenolpyruvate carboxykinase (GTP) causes inactivation without photolabeling

Purpose: To analyze p53, bcl-2, c-myc, and cyclooxygenase-2 protein expression changes and examine their relationship with various antioxidant enzymes in esophageal carcinoma patients. Methods and Materials: Patients in Group 1 underwent transhiatal esophagectomy and those in Group 2 were administered chemoradiotherapy followed by surgery after 4 weeks of neoadjuvant therapy. Results: The relationship analysis among the various protein markers and antioxidant enzymes showed an inverse correlation between bcl-2 and superoxide dismutase/catalase in tumor tissues, irrespective of the treatment arm followed. An important positive association was observed between bcl-2 and reduced glutathione levels in the tumor tissue of patients receiving neoadjuvant therapy. Another apoptosis-modulating marker, c-myc, in the tumor tissue of Group 2 patients showed similar pattern levels (high and low) as that of superoxide dismutase/catalase. The association of ...

2008-02-01

196

Formation of an intramolecular cystine disulfide during the reaction of 8-azidoguanosine 5 prime -triphosphate with cytosolic phosphoenolpyruvate carboxykinase (GTP) causes inactivation without photolabeling

Phosphoenolpyruvate carboxykinase (GTP) (PEPCK) specifically utilizes a guanosine or inosine nucleotide as a substrate, yet it does not share extended sequence homology with other GTP-binding proteins, and the molecular basis for its nucleotide specificity is not understood. In an effort to locate the enzyme's nucleotide-binding site, the authors have studied the interaction of cytosolic PEPCK from rat liver with the photoprobe 8-azidoGTP, which fulfills the criteria of a specific photoaffinity label for PEPCK. The photoprobe binds reversibly to the enzyme prior to modification and at low concentrations causes greater than 60% inactivation-GTP provides nearly complete protection against inactivation by 8-azidoGTP, whereas phosphoenolpyruvate and metal ions provide partial protection. In addition, the photoprobe is a substrate for the enzyme and has a K_m similar to that for GTP. However, the extent of covalent modification ...

197

Enzyme-induced growth of silver nanoparticles studied on single particle level

Phosphoenolpyruvate carboxykinase (GTP) (PEPCK) specifically utilizes a guanosine or inosine nucleotide as a substrate, yet it does not share extended sequence homology with other GTP-binding proteins, and the molecular basis for its nucleotide specificity is not understood. In an effort to locate the enzyme's nucleotide-binding site, the authors have studied the interaction of cytosolic PEPCK from rat liver with the photoprobe 8-azidoGTP, which fulfills the criteria of a specific photoaffinity label for PEPCK. The photoprobe binds reversibly to the enzyme prior to modification and at low concentrations causes greater than 60% inactivation-GTP provides nearly complete protection against inactivation by 8-azidoGTP, whereas phosphoenolpyruvate and metal ions provide partial protection. In addition, the photoprobe is a substrate for the enzyme and has a K{sub m} similar to that for GTP. However, the extent of covalent ...

1989-11-28

198

Enhancement of starch conversion efficiency with free and immobilized pullulanase and alpha-1,4-glucosidase.

Based on their interesting properties, metal nanoparticles show the potential as an analytical tool in electronic (Burmeister et al. 2004), optical (Yguerabide and Yguerabide 1998), and catalytic applications (Liu 2006). Their characteristics depend on the composition, shape, and size of the single particles. These various properties are utilized in many different approaches such as optics, magnetics (Lang et al. 2007), and laser technology (Csaki et al. 2007). We investigated an alternative method for the synthesis of nanoparticles. In this case, an enzyme, horseradish peroxidase, induces a silver deposition and replaces a metal nanoparticle as the reaction seed. Depending on the reaction time, we could obtain particles in a range of few nanometers up to more than 250 nm. For a better understanding of the enzymatic silver deposition process, the silver particles produced by this process were analyzed by SEM, TEM, and atomic force microscopy (AFM) on a single ...

2009-05-15

199

Chemopreventive effects of Furan-2-yl-3-pyridin-2-yl-propenone against 7,12-dimethylbenz[a]anthracene-inducible genotoxicity

Glucoamylase and pullulanase were immobilized on reconstituted bovine-hide collagen membranes using the covalent azide linkage method. A pretanning step was incorporated into the immobilization procedure to enable the support matrix to resist proteolytic activity while accommodating an operating temperature of 50 degrees C. The immobilized glucoamylase and pullulanase activities were 0.91 and 0.022 mg dextrose equivalent (DE) min(-1) cm(-2) of membrane, respectively. Immobilized glucoamylase had a half-life of 50 days while the immobilized pullulanase had a half-life of 7 days. This is a considerably improved stability over that reported by other researchers. The enzymes were studied in their free and immobilized forms on a variety of starch substrates including waxy maize, a material which contains 80% alpha-1-6-glucosidic linkages. Substrate concentrations ranged from 1% to a typical commercial concentration of 30%. Conversion efficiencies of 90-92% DE were ...

1982-02-01

200

Purification and characterization of four keratinases produced by Streptomyces sp. strain 16 in native human foot skin medium

1-Furan-2-yl-3-pyridin-2-yl-propenone (FPP-3) is an anti-inflammatory agent with a propenone moiety and chemically synthesized recently. In this study, we examined the chemopreventive effect of FPP-3 on 7,12-dimethylbenz[a]anthracene (DMBA)-induced genotoxicity in MCF-7 cells. FPP-3 reduced the formation of the DMBA-DNA adduct. DMBA-induced CYP1A1 and CYP1B1 gene expression and enzyme activity were inhibited by FPP-3. It inhibited DMBA-induced aryl hydrocarbon receptor (AhR) transactivation and DMBA-inducible nuclear localization of the AhR. Induction of detoxifying phase II genes by chemopreventive agents represents a coordinated protective response against oxidative stress and neoplastic effects of carcinogens. Transcription factor NF-E2 related factor 2 (Nrf2) regulates antioxidant response element (ARE) of phase II detoxifying and antioxidant enzymes, such as glutathione S-transferase (GST) and NAD(P)H:quinone oxidoreductase (QR). FPP-3 ...

2008-05-01

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201

British Library Electronic Table of Contents (United Kingdom)

Four extracellular keratinases (designated KI, KII, KIII, and KIV) were produced during submerged aerobic cultivation in a medium containing native human foot skin (NHFS) for enzyme synthesis. The molecular weights, determined by SDS-PAGE, were 25, 50, 34, and 19kDa, respectively. Gel filtration of the four purified enzymes in native conditions indicated that active keratinase KI is a novel homo-octamer, KII a homo-dimer, and KIII and KIV monomers. All four keratinases exhibited high activities at pH 8.0-10.0 with an optimal pH of 9.0. The optimal temperature for keratinolytic activity of KI, KII, and KIII was approximately 50, and 60degreeC for KIV. One millimolar of PMSF completely inhibited the keratinolytic activities of the four enzymes. The N-terminal sequences of KI, KII, and KIII s...

2010-01-01

202

Nuclear Targeting of Methyl-Recycling Enzymes in Arabidopsis thaliana Is Mediated by Specific Protein Interactions.

New Insights into the Design of Inhibitors of Human S-Adenosylmethionine Decarboxylase: Studies of Adenine C[superscript 8] Substitution in Structural Analogues of S-Adenosylmethionine

Numerous transmethylation reactions are required for normal plant growth and development. S-adenosylhomocysteine hydrolase (SAHH) and adenosine kinase (ADK) act coordinately to recycle the by-product of these reactions, S-adenosylhomocysteine (SAH) that would otherwise competitively inhibit methyltransferase (MT) activities. Here, we report on investigations to understand how the SAH produced in the nucleus is metabolized by SAHH and ADK. Localization analyses using green fluorescent fusion proteins demonstrated that both enzymes are capable of localizing to the cytoplasm and the nucleus, although no obvious nuclear localization signal was found in their sequences. Deletion analysis revealed that a 41-amino-acid segment of SAHH (Gly(150)-Lys(190)) is required for nuclear targeting of this enzyme. This segment is surface exposed, shows unique sequence conservation patterns in plant SAHHs, and possesses additional features of protein-protein ...

2011-10-01

203

Lipid domain formation modulates activities of snake venom phospholipase A(2) enzymes.

S-Adenosylmethionine decarboxylase (AdoMetDC) is a critical enzyme in the polyamine biosynthetic pathway and depends on a pyruvoyl group for the decarboxylation process. The crystal structures of the enzyme with various inhibitors at the active site have shown that the adenine base of the ligands adopts an unusual syn conformation when bound to the enzyme. To determine whether compounds that favor the syn conformation in solution would be more potent AdoMetDC inhibitors, several series of AdoMet substrate analogues with a variety of substituents at the 8-position of adenine were synthesized and analyzed for their ability to inhibit hAdoMetDC. The biochemical analysis indicated that an 8-methyl substituent resulted in more potent inhibitors, yet most other 8-substitutions provided no benefit over the parent compound. To understand these results, we used computational modeling and X-ray crystallography to study C{sup ...

2009-04-02

204

5,10 Methylenetetrahydrofolate reductase genetic polymorphism as a risk factor for neural tube defects

The goal of the present study is to elucidate the effect of lipid domain formation on activities of Naja naja atra and Bungarus multicinctus phospholipase A(2) (PLA(2)) enzymes. Sphingomyelin inhibited enzymatic activity and membrane-damaging activity of PLA(2) against egg yolk phosphatidylcholine (EYPC), while cholesterol and cholesterol sulfate abrogated the inhibitory effect of sphingomyelin. The ability of cholesterol and cholesterol sulfate to abolish the inhibitory effect of sphingomyelin was closely related to their capacity to induce domain formation in EYPC/sphingomyelin vesicles. Laurdan fluorescence measurement revealed that membrane packing of EYPC/sphingomyelin vesicles was differently affected by cholesterol and cholesterol sulfate. Unlike cholesterol, cholesterol sulfate was unable to promote domain formation in dipalmitoylphosphatidylcholine (DPPC) vesicles. Cholesterol increased but cholesterol sulfate reduced PLA(2) activity against DPPC. ...

2010-08-10

205

The anti-snake venom properties of Tamarindus indica (leguminosae) seed extract

Persons with a thermolabile form of the enzyme 5,10 methylenetetrahydrofolate reductase (MTHFR) have reduced enzyme activity and increased plasma homocysteine which can be lowered by supplemental folic acid. Thermolability of the enzyme has recently been shown to be caused by a common mutation (677C{sup {r_arrow}}T) in the MTHFR gene. We studied 41 fibroblast cultures from NTD-affected fetuses and compared their genotypes with those of 109 blood specimens from individuals in the general population. 677C{sup {r_arrow}}T homozygosity was associated with a 7.2 fold increased risk for NTDs (95% confidence interval: 1.8-30.3; p value: 0.001). These preliminary data suggest that the 677C{sup {r_arrow}}T polymorphism of the MTHFR gene is a risk factor for spina bifida and anencephaly that may provide a partial biologic explanation for why folic acid prevents these types of NTD. 13 refs., 1 fig., 1 tab.

1996-06-28

206

British Library Electronic Table of Contents (United Kingdom)

In Indian traditional medicine, various plants have been used widely as a remedy for treating snakebites. The aim of this study was to evaluate the effect of Tamarindus indica seed extract on the pharmacological as well as the enzymatic effects induced by V. russelli venom. Tamarind seed extract inhibited the PLA2, protease, hyaluronidase, L-amino acid oxidase and 5prime-nucleotidase enzyme activities of venom in a dose-dependent manner. These are the major hydrolytic enzymes responsible for the early effects of envenomation, such as local tissue damage, inflammation and hypotension. Furthermore, the extract neutralized the degradation of the Bb chain of human fibrinogen and indirect hemolysis caused by venom. It was also observed that the extract exerted a moderate effect on the clotting ...

2006-01-01

207

Resistance pattern and antioxidant enzyme profiles of protoporphyrinogen oxidase (PROTOX) inhibitor-resistant transgenic rice

British Library Electronic Table of Contents (United Kingdom)

We quantified the resistance levels of transgenic rice plants, expressing Myxococcus xanthus protoporphyrinogen oxidase (PROTOX) in chloroplasts and mitochondria, to PROTOX inhibitors, acifluorfen, oxyfluorfen, carfentrazone-ethyl, and oxadiazon. We also determined whether active oxygen species-scavenging enzymes are involved in the resistance mechanism of transgenic rice. The transgenic rice line M4 was about >200-fold more resistant to oxyfluorfen than the wild-type (WT). M4 was also resistant to acifluorfen, carfentrazone-ethyl, and oxadiazon, but did not show multiple resistance to imazapyr and paraquat, which have different target sites. Acifluorfen, oxyfluorfen, carfentrazone-ethyl, and oxadiazon reduced the chlorophyll content in leaves of WT, but had minimal or no effect on M4. The...

2008-01-01

208

Removal of aqueous phenol and 2-chlorophenol with purified soybean peroxidase and raw soybean hulls

Glucose Biosensor Based on the Fabrication of Glucose Oxidase in the Bio-Inspired Polydopamine-Gold Nanoparticle Composite Film

The removal phenol and 2-chlorophenol using soybean seed-hulls in the presence of hydrogen peroxide was demonstrated. The performance of a stirred membrane reactor containing soluble purified SEP was compared with a batch stirred reactor containing raw soybean seed-hulls. The purified enzyme reactor proved to be ineffective while good results were obtained with the crude seed-hulls for the removal of phenol and 2-chlorophenol. A single batch reactor containing raw seed-hulls was effective in removing more than 98.5% of 2-chlorophenol (initially at 10000 ppm) in less than 15 min. The performance of these rectors is comparable to existing HRP-based technology. The stability of the soybean peroxidase (SBP) enzyme was also examined in the presence of detergents (SDS, Tween 20 and Triton X-100).

1999-08-01

209

British Library Electronic Table of Contents (United Kingdom)

Abstract A highly efficient enzyme immobilization method has been developed for electrochemical biosensors using polydopamine films with gold nanoparticles (AuNPs) embedded. This simple enzyme fabrication method can be performed in very mild conditions and stored in a long time with high bioactivity. The fabricated amperometric glucose biosensor exhibited a high and reproducible sensitivity, wide linear dynamic range and low limit of detection (LOD) (0.1 molL-1). A low value of 1.5 mmolL-1 for the apparent Michaelis-Menten constant KappM was obtained. The high sensitivity, wide linear range, good reproducibility and stability make this biosensor a promising candidate for portable amperometric glucose biosensor.

2010-01-01

210

From Egg to Crystal: A Practical on Purification, Characterization, and Crystallization of Lysozyme for Bachelor Students

Effect of processing of dates into date juice concentrate and appraisal of its quality characteristics

A practical hands-on course encompassing enzyme purification, biochemical characterization, and crystallization that completed the course work of 350 second-year bachelor students enrolled in molecular biology/biochemistry was given at the Universite Louis Pasteur of Strasbourg (France). The experimental part of the practical dealt entirely with the model protein lysozyme isolated from hen egg-white. It was designed as a research project to give students the possibility to practice biochemical methods such as chromatography, electrophoresis, and spectrophotometry. It also included enzyme activity assay and protein crystal growth that are usually taught in master-level courses. The organization of the practical work and the related experimental procedures are described and discussed.

2007-07-01

211

British Library Electronic Table of Contents (United Kingdom)

Date palm (Phoenix dactylifera) is widely cultivated in Kutch district of Gujarat and the fruits are harvested at immature stage before the onset of monsoon to prevent spoilage. The immature date fruits with less commercial value were used for processing into date juice concentrate. Immature dates were crushed and treated with 0.1% pectinase enzyme for 120 min to obtain maximum juice. Date juice was found to be rich in reducing sugars (16.1%) and total sugars (18.3%). Juice was pasteurized at 85?C to inactivate the enzyme, cooled and centrifuged at 3000 rpm to get clear juice. The juice was concentrated in a thin film evaporator to a total soluble solids (TSS) of 76?Brix in 2 passes. Chemical composition of date juice during different stages of concentration was determined. Date juice conc...

2010-01-01

212

Effect of Tong Luo Jiu Nao on Ab-degrading enzymes in AD rat brains

British Library Electronic Table of Contents (United Kingdom)

Ethnopharmacological relevance: Tong Luo Jiu Nao (TLJN) is a modern Chinese formula based on Traditional Chinese Medicine theory that has been used to treat ischemic cerebral stroke and vascular dementia. TLJN belongs to the ethnopharmacological family of medicines. In this study, we investigated the mechanism of the TLJN effect on Alzheimer's disease (AD). Aim of the study: To investigate the effect of TLJN on b-amyloid-degrading enzymes and learning and memory in the AD rat brain. Materials and methods: AD rats whose disease was induced by Ab25-35 injection into the bilateral hippocampus CA1 region were subjected to intragastric administration of various preparations. The experimental animals were healthy male Sprague-Dawley rats which were randomly divided into normal, sham, model, TLJN...

2011-01-01

213

Comparative biochemistry of betaine biosynthesis and accumulation in diverse dicot families

Chitinolytic enzyme production and genetic improvement of a new isolate belonging to Streptomyces anulatus

Salt stress elicits betaine accumulation to high levels in species from several diverse dicot families (Chenopodiaceae, Amaranthaceae, Convolvulaceae, Solanaceae, and Asteraceae). FAM-MS studies with deuterated precursors showed that species from all these families synthesize betaine from choline. Enzyme assays and immunotitration data showed that all accumulating species contained betaine aldehyde dehydrogenase (BADH) enzyme activity recognized by antibodies raised against purified BADH isolated from Spinacia oleracea. Immunoblotting indicated that the BADH monomer was in all cases of Mr {approx} 63,000. The similarity of BADH in the different species is consistent with a single evolutionary origin for the betaine pathway. This was supported by the presence in immunoblots of a cross-reacting band at Mr {approx} 63,000 in Magnolia x Soulangiana, a primitive angiosperm.

1989-04-01

214

British Library Electronic Table of Contents (United Kingdom)

Thirty bacterial isolates were obtained from different sources and sites at Jeddah, Saudi Arabia, on chitin agar medium; 9 of the 30 isolates were cultured in liquid medium containing chitin as sole carbon and nitrogen sources. Isolate SM21, which was isolated from shrimp shells, showed the best growth and chitinase production in liquid medium. According to its morphological, physiological and biochemical characteristics, SM21 belongs to the genus Streptomyces and was identified as Streptomyces anulatus SM21. Identification was confirmed using 16S rDNA analysis. The chitinase enzyme was precipitated with 80% NH4SO4 and purified using DEAE-cellulose ion exchange chromatography followed by Sephadex G-100 gel filtration. The molecular weight determined using sodium dodecyl sulfate polyacrylam...

2011-01-01

215

Characterization of enzyme-deproteinized skim rubber

British Library Electronic Table of Contents (United Kingdom)

Skim natural rubber latex is a protein-rich byproduct obtained during the centrifugal concentration of natural rubber latex. Skim latex has a very low dry rubber content (4-8%), and the rubber particles are smaller in size. It has a higher proportion of nonrubber solids, which are mostly proteinaceous in nature. It is difficult to coagulate, and it takes more processing time. The proteins in skim latex can be decomposed by proteolytic enzymes. This article discusses the use of stabilized liquid papain from the papaya plant (Carica papaya) for deproteinization followed by creaming for quick and easy coagulation of skim latex. The technological properties and aging characteristics of the deproteinized skim rubber are compared with those of conventionally prepared skim rubber and block rubber...

2009-01-01

216

Bioconvertion of spent cellulose sausage casings

British Library Electronic Table of Contents (United Kingdom)

Cellulose sausage cellulose casings are used extensively in the manufacture of sausages in meat packaging. After stripping the meat, spent casings mainly contain cellulose and residual meat juice with salt, nitrate and nitrite. Disposal of spent sausage casings has serious economic and environmental concerns for the sausage industry. This work describes bioconversion of spent cellulose casings (SCC) into enzymes, lactic acid and ethanol by using cellulolytic fungi, lactobacillus and yeasts. The solid substrate cultivation (SSC) of Trichoderma reesei RUT C-30 on SCC and blends gave a maximum of 152 filter paper cellulase (FPase) activity and about 100 carboxymethylcellulase activity (CMCase)/g dry weight substrate. The SSC produced enzyme-rich casing with 50 FPase when directly mixed as suc...

2008-01-01

217

Ability of a solid state fermentation technique to significantly minimize catabolic repression of. alpha. -amylase production by Bacillus licheniformis M27

A novel structural mechanism for redox regulation of uridine phosphorylase 2 activity

The production of {alpha}-amylase by Bacillus licheniformis M27 in submerged fermentation was completely inhibited due to catabolic repression in medium containing 1% glucose. In contrast, the enzyme production in a solid state fermentation system was 19,550 units/ml extract even when the medium contained 15% glucose. The peak in enzyme titre was, however, shifted from 48 to 72 h. The ability of the solid state fermentation system to significantly overcome catabolic repression was not known earlier and is probably conferred by various physico-chemical factors and culture conditions specific to the system. (orig.).

1991-08-01

218

British Library Electronic Table of Contents (United Kingdom)

Uridine phosphorylase (UPP) catalyzes the reversible conversion of uridine to uracil and ribose-1-phosphate and plays an important pharmacological role in activating fluoropyrimidine nucleoside chemotherapeutic agents such as 5-fluorouracil and capecitabine. Most vertebrate animals, including humans, possess two homologs of this enzyme (UPP1 & UPP2), of which UPP1 has been more thoroughly studied and is better characterized. Here, we report two crystallographic structures of human UPP2 (hUPP2) in distinctly active and inactive conformations. These structures reveal that a conditional intramolecular disulfide bridge can form within the protein that dislocates a critical phosphate-coordinating arginine residue (R100) away from the active site, disabling the enzyme. In vitro activity measurem...

2011-01-01

219

A high-performance liquid chromatography-based radiometric assay for sucrose-phosphate synthase and other UDP-glucose requiring enzymes

uv photobiology: postreplication repair. [Escherichia coli, synchrotron radiation

A method for product analysis that eliminates a problematic step in the radiometric sucrose-phosphate synthase assay is described. The method uses chromatography on a boronate-derivatized high-performance liquid chromatography column to separate the labeled product, (14C)sucrose phosphate, from unreacted uridine 5{prime}-diphosphate-(14C)glucose (UDP-Glc). Direct separation of these compounds eliminates the need for treatment of the reaction mixtures with alkaline phosphatase, thereby avoiding the problem of high background caused by contaminating phosphodiesterase activity in alkaline phosphatase preparations. The method presented in this paper can be applied to many UDP-Glc requiring enzymes; here the authors show its use for determining the activities of sucrose-phosphate synthase, sucrose synthase, and uridine diphosphate-glucose pyrophosphorylase in plant extracts.

1991-05-01

220

Wide cross-species aminoacyl-tRNA synthetase replacement in vivo: yeast cytoplasmic alanine enzyme replaced by human polymyositis serum antigen.

The following topics are discussed: insertion of incorrect bases in DNA; ability of DNA polymerase to copy uv-irradiated DNA; role of enzymes in repair of DNA; effects of uv radiation on molecular weight of DNA; photoreactivation; repair of DNA in Escherichia coli and xp cells following uv radiation; and synchrotron radiation studies on DNA repair. (HLW)

1978-01-01

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221

Variations in Kinetic Properties of Ribulose-1,5-bisphosphate Carboxylases among Plants

Because of variations in tRNA sequences in evolution, tRNA synthetases either do not acylate their cognate tRNAs from other organisms or execute misacylations which can be deleterious in vivo. We report...Full Text Available

1995-05-23

222

The oxygenation of [3-methyl-3H]desacetoxycephalosporin C [7beta-(5-D-aminadipamido)-3-methylceph-3-em-4-carboxylic acid] to [3-hydroxymethyl-3H]desacetylcephalosporin C by 2-oxoglutarate-linked dioxygenases from Acremonium chrysogenum and Streptomyces clavuligerus.

Studies of ribulose-1,5-bisphosphate (RuBP) carboxylase from taxonomically diverse plants show that the enzyme from C₃ and crassulacean acid metabolism pathway species exhibits lower K_m(CO₂)...Full Text Available

1981-06-01

223

The opposite effect of bivalent cations on cytochrome b5 reduction by NADH:cytochrome b5 reductase and NADPH:cytochrome c reductase.

Cell-free extracts of Acremonium chrysogenum and Streptomyces clavuligerus oxidize the 3-methyl group of desacetoxycephalosporin C to a 3-hydroxymethyl group. The enzyme responsible for this reaction...Full Text Available

1978-09-01

224

The mutT Defect Does Not Elevate Chromosomal Fragmentation in Escherichia coli Because of the Surprisingly Low Levels of MutM/MutY-Recognized DNA Modifications?

The effects of bivalent cations on cytochrome b5 reduction by NADH:cytochrome b5 reductase and NADPH:cytochrome c reductase were studied with the proteinase-solubilized enzymes. Cytochrome b5 reduction...Full Text Available

1988-05-01

225

The effect of phytic acid on the levels of blood glucose and some enzymes of carbohydrate and lipid metabolism.

Nucleotide pool sanitizing enzymes Dut (dUTPase), RdgB (dITPase), and MutT (8-oxo-dGTPase) of Escherichia coli hydrolyze noncanonical DNA precursors to prevent incorporation of base...Full Text Available

2007-10-01

226

The Isolation and Characterization of d-Glucose 6-Phosphate Cycloaldolase (NAD-Dependent) from Acer pseudoplatanus L. Cell Cultures

In this study, six groups of rats were fed as follows: Groups 1 and 2 were fed formulated diets supplemented with zinc or without zinc respectively. Groups 3 and 4 were fed formulated diets supplemented with zinc plus phytic acid extracted from sweet potato (Ipomea batatas) or commercial phytic acid respectively. Groups 5 and 6 were fed formulated diets supplemented with phytic acid extract from sweet potato or commercial phytic acid respectively. The animals were fed for three weeks and then sacrificed The activities of key enzymes of carbohydrate and lipid metabolism as well as transaminases in the liver were determined. Blood glucose level was also assessed. Phytic acid extract consumption from sweet potato and commercial phytic acid plus zinc supplement lowered blood glucose levels. There was no significant change in the activity of 6-phosphogluconate dehydrogenase among the groups. Similarly, phytic acid supplementation showed no significant decrease in the ...

2005-03-01

227

The Function of Hypoxia-Inducible Factor (HIF) Is Independent of the Endoplasmic Reticulum Protein OS-9

A soluble enzyme system from suspension cultures of Acer pseudoplatanus L. converts d-glucose 6-phosphate to myoinositol. A Mg²⁺-dependent phosphatase, present in...Full Text Available

1971-09-01

228

The Conversion of Nitrite to Nitrogen Oxide(s) by the Constitutive NAD(P)H-Nitrate Reductase Enzyme from Soybean ¹

The protein “amplified in osteosarcoma-9” (OS-9) has been shown previously to interact with the prolyl hydroxylases PHD2 and PHD3. These enzymes initiate oxygen-dependent degradation...Full Text Available

229

Synthesis and stereochemical effects of pyrrolidinyl-acetylenic thieno[3,2-d]pyrimidines as EGFR and ErbB-2 inhibitors

A two-step purification protocol was used in an attempt to separate the constitutive NAD(P)H-nitrate reductase [NAD(P)H-NR, pH 6.5; EC 1.6.6.2] activity from the nitric oxide and nitrogen dioxide (NO_(x))...Full Text Available

1988-10-01

230

British Library Electronic Table of Contents (United Kingdom)

A novel class of pyrrolidinyl-acetyleneic thieno[3,2-d]pyrimidines has been identified which potently inhibit the EGFR and ErbB-2 receptor tyrosine kinases. Synthetic modifications of the pyrrolidine carbamate moiety result in a range of effects on enzyme and cellular potency. In addition, the impact of the absolute stereochemical configuration on cellular potency and oral mouse pharmacokinetics is described.

2009-01-01

231

Stearoyl-Coenzyme A Desaturase 1 Deficiency Protects against Hypertriglyceridemia and Increases Plasma High-Density Lipoprotein Cholesterol Induced by Liver X Receptor Activation

Quantitative transfer of the molybdenum cofactor from xanthine oxidase and from sulphite oxidase to the deficient enzyme of the nit-1 mutant of Neurospora crassa to yield active nitrate reductase.

Stearoyl-coenzyme A desaturase (SCD) is the rate-limiting enzyme necessary for the biosynthesis of monounsaturated fatty acids. In this study, we investigated the regulation of mouse SCD1 by liver X...Full Text Available

2006-09-01

232

Pre-treatment and ethanol fermentation potential of olive pulp at different dry matter concentrations

An assay method is described for measurement of absolute concentrations of the molybdenum cofactor, based on complementation of the defective nitrate reductase ('apo nitrate reductase') in extracts...Full Text Available

1984-04-15

233

Polyunsaturated Fatty Acids Selectively Suppress Sterol Regulatory Element-binding Protein-1 through Proteolytic Processing and Autoloop Regulatory Circuit*

Renewable energy sources have received increased interest from the international community with biomass being one of the oldest and the most promising ones. In the concept of exploitation of agro-industrial residues, the present study investigates the pre-treatment and ethanol fermentation potential of the olive pulp, which is the semi solid residue generated from the two-phase processing of the olives for olive oil production. Wet oxidation and enzymatic hydrolysis have been applied aiming at the enhancement of carbohydrates' bioavailability. Different concentrations of enzymes and enzymatic durations have been tested. Both wet oxidation and enzymic treatment were evaluated based on the ethanol obtained in a subsequent fermentation step by Saccharomyces cerevisiae and Thermoanaerobacter mathranii. It was found that a four-day hydrolysis time was adequate for a satisfactory release of glucose and xylose. The combination of wet ...

2009-11-15

234

Piperazine sulfonamide BACE1 inhibitors: Design, synthesis, and in vivo characterization

Sterol regulatory element-binding protein (SREBP)-1 is a key transcription factor for the regulation of lipogenic enzyme genes in the liver. Polyunsaturated fatty acids (PUFA) selectively suppress hepatic...Full Text Available

2010-04-09

235

British Library Electronic Table of Contents (United Kingdom)

With collaboration between chemistry, X-ray crystallography, and molecular modeling, we designed and synthesized a series of novel piperazine sulfonamide BACE1 inhibitors. Iterative exploration of the non-prime side and S2prime sub-pocket of the enzyme culminated in identification of an analog that potently lowers peripheral Ab40 in transgenic mice with a single subcutaneous dose.

2010-01-01

236

Molecular identification of CTX-M and ^blaOXY/K1 ?-lactamase genes in Enterobacteriaceae by sequencing of universal M13-sequence tagged PCR-amplicons

Moessbauer studies of the molybdenum-iron-protein in the nitrogenase of Xanthobacter autotrophicus GZ 29. Moessbaueruntersuchungen am Molybdaen-Eisen-Protein der Nitrogenase von Xanthobacter Autotrophicus GZ 29

BackgroundPlasmid encoded ^blaCTX-M enzymes represent an important sub-group of class A β-lactamases causing the ESBL phenotype which is increasingly...Full Text Available

237

Metabolism of 4-chloro-2-methylphenoxyacetate by a soil pseudomonad. Ring-fission, lactonizing and delactonizing enzymes

Biological cells of the nitrogen fixating bacteria Xanthobacter autotrophicus GZ 29 have been cultivated with 57-Fe, and from the nitrogenase of these cells the 57-Fe marked molybdenum-iron-protein is extracted. The Moessbauer spectra of this enzyme show that the structure of the iron cluster is similar to the structure in the molybdenum-iron-proteins of Azotobacter vinelandii, Klebsiella pneumoniae or Clostridium pasteurianum.

1983-05-01

238

Mechanism for the anti-thyroid action of minocycline

1. A cell-free system, prepared from Pseudomonas N.C.I.B. 9340 grown on 4-chloro-2-methylphenoxyacetate (MCPA) was shown to catalyse the reaction sequence: 5-chloro-3-methylcatechol...Full Text Available

1971-05-01

239

Malaysia to release GM mosquitoes into the wild - SciDev.Net

Administration of minocycline (MN), a tetracycline antibiotic, produces a black pigment in the thyroids of humans and several species of experimental animals and antithyroid effects in rodents. We have previously shown that these effects appear to be related to interactions of MN with thyroid peroxidase (TPO), the key enzyme in thyroid hormone synthesis. In the present study, the mechanisms for inhibition of TPO-catalyzed iodination and coupling reactions by MN were investigated. 37 refs., 7 figs., 3 tabs.

1997-01-01

240

Wastenet

... The larvae's only hope for survival is if the antibiotic tetracycline is present -because it mops up the enzyme. The developers hope male GM mosquitoes will compete with normal males for females so that repeated releases cut numbers of A. aegypti in dengue-prone areas. The mosquitoes will be released in the inland districts of Bentong in the state of Pahang, and Alor Gajah and Melaka in the state of Malacca, according to the National Biosafety Board. Each location will have two release phases: the first at a site ...

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241

Locked nucleic acid modified DNA enzymes targeting early growth response-1 inhibit human vascular smooth muscle cell growth

Local and systemic antibody response to bovine respiratory syncytial virus infection and reinfection in calves with and without maternal antibodies.

Smooth muscle cell (SMC) proliferation and migration are key processes that occur in the pathogenesis of atherosclerosis and post-angioplasty restenosis. In the present study, we designed locked nucleic...Full Text Available

2004-01-01

242

Legionella pneumophila serogroup Lansing 3 isolated from a patient with fatal pneumonia, and descriptions of L. pneumophila subsp. pneumophila subsp. nov., L. pneumophila subsp. fraseri subsp. nov., and L. pneumophila subsp. pascullei subsp. nov.

Enzyme-linked immunosorbent assays for the detection of immunoglobulin M (IgM), IgA, IgG1, and IgG2 antibodies against bovine respiratory syncytial virus (BRSV) were used to measure antibody responses...Full Text Available

1987-06-01

243

Isolation and Initial Characterization of Constitutive Nitrate Reductase-Deficient Mutants NR328 and NR345 of Soybean (Glycine max) ¹

Previous DNA relatedness and enzyme electrophoretic mobility studies indicated heterogeneity among strains of Legionella pneumophila serogroups 1, 4, 5, and Lansing 3 (a new, as yet unnumbered serogroup)....Full Text Available

1988-09-01

244

Interactions between tRNA identity nucleotides and their recognition sites in glutaminyl-tRNA synthetase determine the cognate amino acid affinity of the enzyme.

Two nitrate reductase deficient mutants of soybean (Glycine max [L.] Merr. cv Bragg) were isolated from approximately 10,000 M₂ seedlings, using a direct enzymic assay in...Full Text Available

1986-06-01

245

GM mosquito wild release takes campaigners by surprise - SciDev.Net

Sequence-specific interactions between aminoacyl-tRNA synthetases and their cognate tRNAs both ensure accurate RNA recognition and prevent the binding of noncognate substrates. Here we show for Escherichia...Full Text Available

1996-07-09

246

Wastenet

... The male GM mosquitoes mate with normal females to produce larvae that die unless the antibiotic tetracycline is present. In tetracycline's absence an enzyme accumulates to a toxic level, killing the larvae. The developers hope the strategy could be combined with other mosquito control methods to reduce transmission in dengue-prone areas. Ricarda Steinbrecher, a geneticist and co-director of EcoNexus -a UK-based non-profit research organisation -expressed surprise ...

247

GM Mosquito Wild Release Takes Campaigners by Surprise

Wastenet

... The male GM mosquitoes mate with normal females to produce larvae that die unless the antibiotic tetracycline is present. In tetracycline's absence an enzyme accumulates to a toxic level, killing the larvae. The developers hope the strategy could be combined with other mosquito control methods to reduce transmission in dengue-prone areas. Ricarda Steinbrecher, a geneticist and co-director of EcoNexus - a UK-based non-profit research organization - ...

248

Fusion of GFP to the M.EcoKI DNA methyltransferase produces a new probe of Type I DNA restriction and modification enzymes

Functional and Topological Analysis of Yeast Acyl-CoA:Diacylglycerol Acyltransferase 2, an Endoplasmic Reticulum Enzyme Essential for Triacylglycerol Biosynthesis*

AbstractWe describe the fusion of enhanced green fluorescent protein to the C-terminus of the HsdS DNA sequence-specificity subunit of the Type I DNA modification methyltransferase M.EcoKI....Full Text Available

2010-07-23

249

Extended Spectrum ?-Lactamases among Gram-negative bacteria of nosocomial origin from an Intensive Care Unit of a tertiary health facility in Tanzania

Acyl-CoA:diacylglycerol acyltransferase (EC 2.3.1.20) is a membrane protein present mainly in the endoplasmic reticulum. It catalyzes the final and committed step in the biosynthesis of triacylglycerol,...Full Text Available

2011-04-15

250

Exploiting the beneficial effects of PPO on the utilisation of protein and lipids in grazed forages

BackgroundResistance to third generation cephalosporins due to acquisition and expression of extended spectrum β-lactamase (ESBL) enzymes among Gram-negative bacteria is...Full Text Available

251

Environmental Research Database

DescriptionThe plant enzyme polyphenol oxidase (PPO) has been demonstrated to reduce both proteolysis and lipolysis during the ensiling of red clover. The purpose of this work is to explore the potential for exploiting this beneficial trait in other dietary regimes for grazing ruminants. These studies will focus on the development of new strategies for forage management and livestock feeding by increasing the efficiency of feed protein-N utilisation and reducing saturated fatty acid production by ruminan [continued...

2007-01-31

252

Development of species-specific enzyme-linked immunosorbent assay for diagnosis of Johne's disease in cattle.

Contribution of Gene Amplification to Evolution of Increased Antibiotic Resistance in Salmonella typhimurium

The previously described (M. De Kesel, P. Gilot, M.-C. Misonne, M. Coene, and C. Cocito, J. Clin. Microbiol., 31:947-954, 1993) a362 recombinant polypeptide of Mycobacterium paratuberculosis was used...Full Text Available

1994-05-01

253

Comparison between NO_x Evolution Mechanisms of Wild-Type and nr₁ Mutant Soybean Leaves ¹

The use of β-lactam antibiotics has led to the evolution and global spread of a variety of resistance mechanisms, including β-lactamases, a group of enzymes that degrade the β-lactam...Full Text Available

2009-08-01

254

The nr₁ soybean (Glycine max [L.] Merr.) mutant does not contain the two constitutive nitrate reductases, one of which is responsible for enzymic conversion of nitrite to...Full Text Available

1990-05-01

255

Catalytic chemistry

Biochemical and Structural Characterization of the Secreted Chorismate Mutase (Rv1885c) from Mycobacterium tuberculosis H₃₇R_v: an *AroQ Enzyme Not Regulated by the Aromatic Amino Acids

Most chemical reactions in industry and biology are catalytic and play a role at some stage of the processing of about 80% of the goods manufactured in the U.S., yet catalysis is a neglected subject in chemical education. This book integrates the fragmentary treatment accorded the topic until now. It covers, in a unified way, catalysis in solutions, by enzymes, in synthetic polymers within the molecular scale cages of zeolites and other molecular sieves, and on surfaces of inorganic solids. The central ideas are chemical; and principles are illustrated by emphasizing industrial reactions and catalysts.

1991-01-01

256

Alterations in Adenylate Kinase Activity in Human PBMCs after In Vitro Exposure to Electromagnetic Field: Comparison between Extremely Low Frequency Electromagnetic Field (ELF) and Therapeutic Application of a Musically Modulated Electromagnetic Field (TAMMEF)

The gene Rv1885c from the genome of Mycobacterium tuberculosis H₃₇R_v encodes a monofunctional and secreted chorismate mutase (*MtCM) with a 33-amino-acid...Full Text Available

2006-12-01

257

A New Metal-Binding Site for Yeast Phosphoglycerate Kinase as Determined by the Use of a Metal-ATP Analog

This study investigated the effects of electromagnetic fields on enzymes involved in purine metabolism in human peripheral blood mononuclear cells in vitro. Cells were obtained from 20 volunteers. We...Full Text Available

2009-01-01

258

A Combinatorial Interplay Among the 1-Aminocyclopropane-1-Carboxylate Isoforms Regulates Ethylene Biosynthesis in Arabidopsis thaliana

Suicide substrate β, γ-bidentate Rh(III)ATP (RhATP) was used to map the metal ion-binding site in yeast phosphoglycerate kinase (PGK). Cleavage of the RhATP-inactivated enzyme with...Full Text Available

1997-02-01

259

$gamma$-RAY PROCESSING OF RICE. (IV.) STUDIES ON THE AVAILABILITY OF $gamma$-IRRADIATED RICE FOR KOJI

Ethylene (C₂H₄) is a unique plant-signaling molecule that regulates numerous developmental processes. The key enzyme in the two-step biosynthetic pathway of ethylene is 1-aminocyclopropane-1-carboxylate...Full Text Available

2009-11-01

260

The Structure of Sucrose Phosphate Synthase from Halothermothrix orenii Reveals Its Mechanism of Action and Binding Mode

The availability of rice, gamma -irradiated up to 4.6 x 10/sup 4/ and 3.5 x 10/sup 5/r for koji was studied. The enzyme activities of koji of the steamed samples were stronger than the unirradiated rice in amylase and protease. The sensory test on the once-steamed irradiated rice was almost the same as the twice-steamed unirradiated rice. (OID)

1961-07-01

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261

Systems Biology Approach in Chlamydomonas Reveals Connections between Copper Nutrition and Multiple Metabolic Steps[C][W][OA

Sucrose phosphate synthase (SPS) catalyzes the transfer of a glycosyl group from an activated donor sugar, such as uridine diphosphate glucose (UDP-Glc), to a saccharide acceptor D-fructose 6-phosphate (F6P), resulting in the formation of UDP and D-sucrose-6'-phosphate (S6P). This is a central regulatory process in the production of sucrose in plants, cyanobacteria, and proteobacteria. Here, we report the crystal structure of SPS from the nonphotosynthetic bacterium Halothermothrix orenii and its complexes with the substrate F6P and the product S6P. SPS has two distinct Rossmann-fold domains with a large substrate binding cleft at the interdomain interface. Structures of two complexes show that both the substrate F6P and the product S6P bind to the A-domain of SPS. Based on comparative analysis of the SPS structure with other related enzymes, the donor substrate, nucleotide diphosphate glucose, binds to the B-domain of SPS. Furthermore, we propose a ...

2008-01-01

262

Synthesis of alkylated deoxyno irimycin and 1,5-dideoxy-1,5-iminoxylitol analogues: : Polar side-chain modification, sulfonium and selenonium heteroatom variants, conformational analysis, and evaluation as glycosidase inhibitors

In this work, we query the Chlamydomonas reinhardtii copper regulon at a whole-genome level. Our RNA-Seq data simulation and analysis pipeline validated a 2-fold cutoff and 10 RPKM (reads per kilobase of mappable length per million mapped reads) (~1 mRNA per cell) to reveal 63 CRR1 targets plus another 86 copper-responsive genes. Proteomic and immunoblot analyses captured 25% of the corresponding proteins, whose abundance was also dependent on copper nutrition, validating transcriptional regulation as a major control mechanism for copper signaling in Chlamydomonas. The impact of copper deficiency on the expression of several O2-dependent enzymes included steps in lipid modification pathways. Quantitative lipid profiles indicated increased polyunsaturation of fatty acids on thylakoid membrane digalactosyldiglycerides, indicating a global impact of copper deficiency on the photosynthetic apparatus. Discovery of a putative plastid copper chaperone and a membrane ...

2011-04-01

263

Radiolabeling of a wound-inducible pyridoxal phosphate utilizing protein from tomato: evidence for its identification as ACC synthase

The syntheses of N-alkylated deoxynojirimycin and 1,5-dideoxy-1,5-iminoxylitol derivatives having either a D- or an L-erythritol-3-sulfate functionalized N-substituent are reported. The alkylating agent used was a cyclic sulfate derivative, whereby selective attack of the nitrogen atom at the least hindered primary center afforded the desired ammonium salt. In aqueous solution, these salts were configurationally labile at the ammonium center. Sulfonium and/or selenonium analogues of the ammonium salts were prepared by analogous reactions. The chalcogen salts were obtained as mixtures of diastereomers, separable in some cases, differing only in the stereochemistry at the configurationally stable sulfur or selenium atoms. Proof of configuration and conformation of each compound was obtained by detailed NMR experiments. The compounds are six-membered ring analogues of salacinol, a known sulfonium-salt glucosidase inhibitor. Evaluation of the target compounds for ...

2004-01-01

264

Radiation-induced changes in the cell membrane of cultured human endothelial cells

Aminocyclopropane 1-carboxylic acid (ACC) synthase, a pyridoxal phosphate utilizing enzyme, catalyzes the conversion of S-adenosylmethionine to ACC, the rate limiting step in the biosynthesis of the plant hormone, ethylene. Ethylene, besides being involved in normal plant growth processes, is also produced in response to stress, e.g. wounding, pathogen infection, etc. The authors report the partial purification (400 fold) of ACC synthase from wounded pink tomato pericarp by classical techniques including ammonium sulfate precipitation, ion exchange and phenyl sepharose chromatography. Further purification results in a decrease in specific activity apparently due to the instability of the enzyme and the low levels present in plant tissue. Radiolabeling of a pyridoxal phosphate-utilizing protein in the ACC synthase enriched fraction was achieved. Evidence that this radiolabeled protein is ACC synthase will be presented. Amino acid sequence ...

1986-06-08

265

Pa0148 from Pseudomonas aeruginosa Catalyzes the Deamination of Adenine.

We investigated the effect of irradiation on the kinetic characteristics of amino acid and glucose transport, and the effect on the activity of the cell membrane-bound enzyme 5'-nucleotidase and on the receptor-mediated stimulation of cyclic adenosine monophosphate synthesis by prostaglandin E1. Irradiation inhibited the sodium-dependent amino acid transport by a reduced binding of the amino acid to the transport unit. The transport of glucose, which appeared to be a sodium-independent process, was temporarily stimulated by increased maximal velocity of the transport. No effect was found on the binding to the transport unit. Irradiation increased the 5'-nucleotidase activity and decreased the prostaglandin E1-stimulated cyclic adenosine monophosphate synthesis 48 h after exposure to 20 Gy. It is concluded that irradiation decreases sodium-dependent transport by impairment of the transport unit, does not impair a sodium-independent process, and has ...

1985-12-01

266

Origin and spread of a common deletion causing mucolipidosis type II: insights from patterns of haplotypic diversity

Four proteins from NCBI cog1816, previously annotated as adenosine deaminases, have been subjected to structural and functional characterization. Pa0148 (Pseudomonas aeruginosa PAO1), AAur1117 (Arthrobacter aurescens TC1), Sgx9403e, and Sgx9403g have been purified and their substrate profiles determined. Adenosine is not a substrate for any of these enzymes. All of these proteins will deaminate adenine to produce hypoxanthine with k(cat)/K(m) values that exceed 10(5) M(-1) s(-1). These enzymes will also accept 6-chloropurine, 6-methoxypurine, N-6-methyladenine, and 2,6-diaminopurine as alternate substrates. X-ray structures of Pa0148 and AAur1117 have been determined and reveal nearly identical distorted (?/?)(8) barrels with a single zinc ion that is characteristic of members of the amidohydrolase superfamily. Structures of Pa0148 with adenine, 6-chloropurine, and hypoxanthine were also determined, thereby permitting identification of the ...

2011-07-06

267

British Library Electronic Table of Contents (United Kingdom)

Coutinho MF, Encarnao M, Gomes R, da Silva Santos L, Martins S, Sirois-Gagnon D, Bargal R, Filocamo M, Raas-Rothschild A, Tappino B, Laprise C, Cury GK, Schwartz IV, Artigals O, Prata MJ, Alves S. Origin and spread of a common deletion causing mucolipidosis type II: insights from patterns of haplotypic diversity. Mucolipidosis II (ML II alpha/beta), or I-cell disease, is a rare genetic disease in which activity of the uridine diphosphate (UDP)-N-acetylglucosamine:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase (GlcNAc-phosphotransferase) is absent. GlcNAc-phosphotransferase is a multimeric enzyme encoded by two genes, GNPTAB and GNPTG. A spectrum of mutations in GNPTAB has been recently reported to cause ML II alpha/beta. Most of these mutations were found to be private or rare. ...

2011-01-01

268

Lipase catalyzed esterification of glycidol in organic solvents

Investigation of enzymatic biodiesel production using ionic liquid as a co-solvent

The authors studied the resolution of racemic glycidol through esterification with butyric acid catalyzed by porcine pancreatic lipase in organic media. A screening of seven solvents (log P values between 0.49 and 3.0, P being the n-octanol-water partition coefficient of the solvent) showed that neither log P nor the logarithm of the molar solubility of water in the solvent provides good correlations between enantioselectivity and the properties of the organic media. Chloroform was one of the best solvents as regards the enantiometic purity (e.p.) of the ester produced. In this solvent, the optimum temperature for the reaction was determined to be 35C. The enzyme exhibited maximum activity at a water content of 13 [plus minus] 2% (w/w). The enantiomeric purity obtained was 83 [plus minus] 2% of (S)-glycidol butyrate and did not depend on the alcohol concentration or the enzyme water content for values of these parameters up to 200 mM and 25% ...

1993-08-05

269

Effect of pentachlorophenol on the activation of 2,6-dinitrotoluene to genotoxic urinary metabolites in CD-1 mice: A comparison of G1 enzyme activities and urine mutagenicity

Ionic liquids can enhance the activity, selectivity, and stability of enzyme catalysts. Enzymes and ionic liquids as catalysts are seen as a more environmentally friendly technology than the use of traditional catalyst materials in biofuels production. This study investigated the lipase catalyzed esterification reaction for biodiesel production in the presence of the ionic liquid BMIM PF{sub 6}. Methyl acetate was used as the acyl acceptor. A titremetric assay was conducted to measure the activity of the lipase with various amounts of ionic liquid. Reaction parameters were investigated in order to determine their effects on biodiesel conversion. An 80 per cent overall methyl oleate biodiesel yield was successfully produced in the presence of the ionic liquid. The ionic liquid improved lipase activity as well as overall biodiesel yield. Results also showed that the addition of the ionic liquid facilitated the separation of methyl esters from ...

2010-04-15

270

Crystal structures of two novel sulfonylurea herbicides in complex with Arabidopsis thaliana acetohydroxyacid synthase

2,6-Dinitrotoluene (2,6-DNT) and pentachlorophenol (PCP) are used for industrial purposes and are found in the environment as hazardous contaminants. Because concurrent exposure to both compounds can occur, it is of interest to determine if organochlorine compounds potentiate the effect of nitroaromatic chemicals. A significant increase in mutagenicity was observed in urines from mice treated with 2,6-DNT alone and in combination with PCP. By week 4, mice that received both 2,6-DNT and PCP excreted urine that was more mutagenic than that from animals which received only 2,6-DNT. At weeks 2 and 4, mice were sacrificed and intestinal enzyme activities (nitroreductase, azo reductase, {beta}-glucuronidase, dechlorinase, and dehydrochlorinase) were quantitated. The enhanced genotoxicity observed in urines from 2,6-DNT/PCP-treated mice coincided with a decrease in nitroreductase and an increase in {beta}-glucuronidase activities in the small intestine.

1991-01-01

271

Changes in yogurt fermentation characteristics, and antioxidant potential and in vitro inhibition of angiotensin-1 converting enzyme upon the inclusion of peppermint, dill and basil

Acetohydroxyacid synthase (AHAS; EC 2.2.1.6) is the first enzyme in the biosynthetic pathway of the branched-chain amino acids. It catalyzes the conversion of two molecules of pyruvate into 2-acetolactate or one molecule of pyruvate and one molecule of 2-ketobutyrate into 2-aceto-2-hydroxybutyrate. AHAS requires the cofactors thiamine diphosphate (ThDP), Mg{sup 2+} and FAD for activity. The herbicides that target this enzyme are effective in protecting a broad range of crops from weed species. However, resistance in the field is now a serious problem worldwide. To address this, two new sulfonylureas, monosulfuron and monosulfuron ester, have been developed as commercial herbicides in China. These molecules differ from the traditional sulfonylureas in that the heterocyclic ring attached to the nitrogen atom of the sulfonylurea bridge is monosubstituted rather than disubstituted. The structures of these compounds in complex with the catalytic ...

2009-08-17

272

British Library Electronic Table of Contents (United Kingdom)

The present study investigated the effects of peppermint (Mentha piperita), dill (Anethum graveolens) and basil (Ocimum basilicum) on yogurt formation, proteolysis and inhibition of angiotensin-1 converting enzyme (ACE). Herbal-yogurts had faster rates of pH reduction than plain-yogurt. All herbal-yogurts had higher (p < 0.05) antioxidant activities than plain-yogurt, both at the end of fermentation and throughout the storage period. The o-phthalaldehyde (OPA) peptides in herbal-yogurts increased by 28-36% after 7 days of storage. All herbal-yogurts showed higher anti-ACE activity than plain-yogurt at corresponding storage periods. M. piperita yogurt had highest inhibitory effect on ACE activity throughout the storage period. Peppermint, dill and basil may be used to modify microbial ferme...

2011-01-01

273

The effect of mitochondrial dysfunction on cytosolic nucleotide metabolism

Osmoregulation in methanogens. Progress report, May 15, 1991--January 15, 1993

Several enzymes of the metabolic pathways responsible for metabolism of cytosolic ribonucleotides and deoxyribonucleotides are located in mitochondria. Studies described in this paper suggest dysfunction of the mitochondria to affect these metabolic pathways and limit the available levels of cytosolic ribonucleotides and deoxyribonucleotides, which in turn can result in aberrant RNA and DNA synthesis. Mitochondrial dysfunction has been linked to genomic instability, and it is possible that the limiting effect of mitochondrial dysfunction on the levels of nucleotides and resulting aberrant RNA and DNA synthesis in part can be responsible for this link. This paper summarizes the parts of the metabolic pathways responsible for nucleotide metabolism that can be affected by mitochondrial dysfunction.

2010-01-01

274

Osmoregulation in methanogens

Our major goal of our work has been to develop and use NMR techniques to study how methanogenic archaebacteria deal with osmotic stress with the hope of providing insights into increasing the salt tolerance of other cells. The project has three main sections: (i) in vivo studies of methanogens; (ii) use of {sup l3}C- and {sup l5}N- labeled potential precursors and in vitro analyses of specific label uptake for elucidation of osmolyte dynamics and biosynthetic pathways of osmolytes in these organisms, and isolation of key biosynthetic enzymes; and (iii) collaborative studies on identification of organic solutes in other methanogens.

1993-01-01

275

Metabolism of benzo(a)pyrene by isolated perfused testis and testicular homogenate

Our major goal of our work has been to develop and use NMR techniques to study how methanogenic archaebacteria deal with osmotic stress with the hope of providing insights into increasing the salt tolerance of other cells. The project has three main sections: (i) in vivo studies of methanogens; (ii) use of [sup l3]C- and [sup l5]N- labeled potential precursors and in vitro analyses of specific label uptake for elucidation of osmolyte dynamics and biosynthetic pathways of osmolytes in these organisms, and isolation of key biosynthetic enzymes; and (iii) collaborative studies on identification of organic solutes in other methanogens.

1993-01-01

276

In vivo recovery of glycogen metabolism in hemolymph and tissues of a freshwater field crab Barytelphusa guerini on exposure to hexavalent chromium

In an effort to improve the extrapolation of laboratory data to man and estimate risk of human reproductive toxicity associated with environmental exposure, the pharmacokinetic parameters of the testicular compartment are being studied. Of particular interest is the variety of enzyme systems capable of activating and detoxicating environmental chemicals and drugs. This report compares the metabolism of benzo(a)pyrene by the isolated perfused testis and testicular homogenates in vitro. The cell free in vitro system metabolized benzo(a)pyrene at a much greater rate than the perfused testis and produced a different spectrum of metabolites. Reliable laboratory prediction of biotransformation by the whole organ or intact animal is an essential aspect of reproductive toxicology.

1980-12-01

277

In vitro evaluation of herpes simplex virus type 1 thymidine kinase reporter system in dynamic studies of transcriptional gene regulation

The in vivo toxic effects of hexavalent chromium (20 mg/liter) on hemolymph glucose, tissue glycogen, total free sugars, and active and total phosphorylases of an edible, freshwater crab Barytelphusa guerini were studied. In a 15-day exposure span followed by a 15-day postexposure recovery, the time-course alterations in these constituent segments of the glycogen metabolism indicate an inconsistent depletion in metabolite levels and elevated enzyme activities during exposure period as well as hyperglycemia. An insignificant recovery was observed in these parameters on the 15th day of the postexposure phase.

1990-08-01

278

Hypersersensitivity and Kounis syndrome due to a viper bite

The herpes simplex virus type 1 thymidine kinase (HSV1-TK) reporter system is being used to directly and indirectly monitor therapeutic gene expression, immune cell trafficking and protein-protein interactions in various living animals. However, the issues of HSV1-TK enzyme stability in living cells and whether this reporter system is optimal for dynamic studies of gene expression events in genetic imaging have not be addressed. The purpose of the present study was to evaluate the application of this reporter system in dynamic studies of transcriptional gene regulation. To achieve this purpose, we established two tetracycline-inducible murine sarcoma cell lines, tetracycline-turn-off HSV1-tk-expressing cell line (NG4TL4/tet-off-HSV1-tk) and tetracycline-turn-off Luc-expressing cell line (NG4TL4/tet-off-Luc), to create an artificially regulated gene expression model in vitro. The dynamic transcriptional events mediating a series of doxycycline (Dox) inductions were ...

2006-07-15

279

British Library Electronic Table of Contents (United Kingdom)

A 60-year-old male was bitten by a venomous snake (Vipera ammodytes) and gradually developed signs of an allergic reaction including generalized itching, generalized rash, and chest discomfort. This was followed by severe retrosternal pain with electrocardiographic evidence of an inferior myocardial ischemia progressing to acute myocardial infarction. Cardiac enzymes and troponin, serum tryptase, and histamine were elevated. Coronary arteriography showed normal coronary arteries. This is a characteristic type I variant of Kounis syndrome, which is the concurrence of acute coronary syndromes with conditions associated with mast cell activation including allergic or hypersensitivity reactions as well as anaphylactic or anaphylactoid reactions. This is the first report to show that viper bite...

2006-01-01

280

Dipyrone-induced changes in DNA repair and other cell membrane associated processes in Escherichia coli

Dipyrone-induced changes in DNA repair and other cell membrane associated processes in Escherichia coli

The analgesic, dipyrone (1,phenyl-2,3-dimethyl-5-pyrazolone-4-methylamino methane sulphonate sodium), at 20 mM concentration, inhibited the rejoining of single-strand scissions in DNA of Escherichia coli B/r cells induced by 20 krad gamma-radiation. The chemical altered the cell membrane structure as evidenced from the uptake of acriflavin, the efflux of potassium ions from the bacterial cells and the inhibition of alkaline phosphatase-a cell membrane associated enzyme. (author). 18 refs., 6 figures.

1982-09-01

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281

British Library Electronic Table of Contents (United Kingdom)

The analgesic, dipyrone (1,phenyl-2,3-dimethyl-5-pyrazolone-4-methylamino methane sulphonate sodium), at 20 mM concentration, inhibited the rejoining of single-strand scissions in DNA of Escherichia coli B/r cells induced by 20 krad gamma-radiation. The chemical altered the cell membrane structure as evidenced from the uptake of acriflavin, the efflux of potassium ions from the bacterial cells and the inhibition of alkaline phosphatase-a cell membrane associated enzyme. (author).

282

Cleavable surfactants

The review covers the development within the field of cleavable surfactants since 2003. Cleavable surfactants are amphiphiles in which a weak linkage has been deliberately inserted, normally, but not always, between the hydrophobic tail and the polar headgroup. Alkali labile linkages that have been used for the purpose include normal ester bonds, betaine esters, and carbonates. Ketals and ortho esters are example of bonds that are susceptible to acid hydrolysis. Several investigations deal with cationic ester-containing surfactants, both monomeric and dimeric species, the latter being gemini surfactants. Ester, amide and carbonate containing surfactants have been investigated with respect to enzyme catalyzed hydrolysis. The main incentive for the development of novel cleavable surfactants ...

2007-01-01

283

Azidothymidine: crystal structure and possible functional role of the azido group

The CYP2A3 gene product catalyzes coumarin 7-hydroxylation in human liver microsomes

The crystal and molecular structures of the anti-acquired immunodeficiency syndrome agent 3'-azido-3'-deoxythymidine (AZT) have been determined by x-ray diffraction. There are two crystallographically independent AZT molecules in the crystal asymmetric unit; they have similar conformation and differ primarily in the glycosyl torsion angle. Comparisons with a hydrated thymidylate structure indicate that the azido group does not significantly affect the gross conformational preference of the molecule. The comparisons also suggest possible functional roles for the azido group in enzyme binding.

1987-12-01

284

The CYP2A3 gene product catalyzes coumarin 7-hydroxylation in human liver microsomes

Three cDNAs, designated IIA3, IIA3v, and IIA4, coding for P450s in the CYP2A gene subfamily were isolated from a {lambda}gt11 library prepared from human hepatic mRNA. Only three nucleotide differences and a single amino acid difference, Leu{sup 160}{yields}His, were found between IIA3 and IIA3v, indicating that they are probably allelic variants. IIA4 displayed 94% amino acid similarity with IIA3 and IIA3v. The three cDNAs were inserted into vaccinia virus, and recombinant viruses were used to infect human hepatoma Hep G2 cells. Only IIA3 was able to produce an enzyme that had a reduced CO-bound spectrum with a {lambda}{sub max} at 450 nm. This expressed enzyme was able to carry out coumarin 7-hydroxylation and ethoxycoumarin O-deethylation. cDNA-expressed IIA3v and IIA4 failed to incorporate heme and were enzymatically inactive. Analysis of IIA proteins in human liver microsomes, using antibody against rat IIA2, revealed two proteins of 49 ...

1990-02-06

285

Regulation of sulfated glycosaminoglycan production by prostaglandin E2 in cultured lung fibroblasts

Three cDNAs, designated IIA3, IIA3v, and IIA4, coding for P450s in the CYP2A gene subfamily were isolated from a #lambda#gt11 library prepared from human hepatic mRNA. Only three nucleotide differences and a single amino acid difference, Leu"1"6"0#->#His, were found between IIA3 and IIA3v, indicating that they are probably allelic variants. IIA4 displayed 94% amino acid similarity with IIA3 and IIA3v. The three cDNAs were inserted into vaccinia virus, and recombinant viruses were used to infect human hepatoma Hep G2 cells. Only IIA3 was able to produce an enzyme that had a reduced CO-bound spectrum with a #lambda#_m_a_x at 450 nm. This expressed enzyme was able to carry out coumarin 7-hydroxylation and ethoxycoumarin O-deethylation. cDNA-expressed IIA3v and IIA4 failed to incorporate heme and were enzymatically inactive. Analysis of IIA proteins in human liver microsomes, using antibody against rat IIA2, revealed two proteins of 49 and 50 ...

1990-02-01

286

One-step purification of Taq DNA polymerase using nucleotide-mimetic affinity chromatography.

Prostaglandin E2 (PGE2) has been shown to increase the synthesis of hyaluronic acid in cultured fibroblasts by increasing the activity of hyaluronate synthetase, a group of plasma membrane-bound synthetic enzymes. We examined whether PGE2 also increased the activity of those enzyme systems involved in the synthesis of sulfated glycosaminoglycan in the human embryonic lung fibroblast. Exposure of cells to PGE2 resulted in dose-dependent increases in glucosamine incorporation into all sulfated glycosaminoglycan subtypes. PGE2 at 10(-7) mol/L increased total glycosaminoglycan per dish to 21.6 +/- 3.1 micrograms versus 12.0 +/- 2.5 micrograms in control untreated cultures. Stimulation of endogenous PGE2 production by bradykinin had a similar effect on glycosaminoglycan synthesis. To examine whether PGE2 affected sulfated glycosaminoglycan protein core production, cells were labeled with tritiated glucosamine in the presence of cycloheximide. Under ...

1989-08-01

287

Increased oxidative stress and antioxidant expression in mouse keratinocytes following exposure to paraquat

The thermostable Thermus aquaticus DNA polymerase (Taq Pol) has been the key factor in transforming the initial PCR method into one with huge impact in molecular biology and biotechnology. Therefore, the development of effective affinity adsorbents for the purification of Taq Pol, as well as other DNA polymerases, attracts the attention of the enzyme manufacturers and the research laboratories. In this report we describe a simple protocol for the purification of Taq Pol from E. coli lysates, leading to enzymes of high specific activity and purity. The protocol is based on a single affinity chromatography step, featuring an immobilized ligand selected from a structure-biased combinatorial library of dNTP-mimetic synthetic ligands. The ligand library was screened for its ability to bind and purify Taq Pol from E. coli lysates. One immobilized ligand (mABSGu) of the general formula X-Trz-Y, bearing 9-aminoethylguanine (AEGu) and aniline-2-sulfonic ...

2007-01-01

288

Enzyme induction in neonates after fetal exposure to antiepileptic drugs

Paraquat (1,1'-dimethyl-4,4'-bipyridinium) is a widely used herbicide known to induce skin toxicity. This is thought to be due to oxidative stress resulting from the generation of cytotoxic reactive oxygen intermediates (ROI) during paraquat redox cycling. The skin contains a diverse array of antioxidant enzymes which protect against oxidative stress including superoxide dismutase (SOD), catalase, glutathione peroxidase-1 (GPx-1), heme oxygenase-1 (HO-1), metallothionein-2 (MT-2), and glutathione-S-transferases (GST). In the present studies we compared paraquat redox cycling in primary cultures of undifferentiated and differentiated mouse keratinocytes and determined if this was associated with oxidative stress and altered expression of antioxidant enzymes. We found that paraquat readily undergoes redox cycling in both undifferentiated and differentiated keratinocytes, generating superoxide anion and hydrogen peroxide as well as increased ...

2008-09-15

289

Cysteinyl peptides of rabbit muscle pyruvate kinase labeled by the affinity label 8-((4-bromo-2,3-dioxobutyl)thio)adenosine 5 prime -triphosphate

The /sup 13/C-AP breath test is shown to be a convenient, noninvasive method to monitor velocity and capacity of P450-dependent AP N-demethylation in infancy and childhood. According to /sup 13/C-AP breath tests, neonates have a very low capacity to eliminate /sup 13/CO/sub 2/, which is only 15 to 21% of the activity in adults. During the first year of life AP N-demethylation increases to reach its maximum at about 2 years; afterwards a slight decrease occurs. In 25 neonates exposed prenatally to different antiepileptic drugs /sup 13/C-AP breath test was efficiently used to prove that cytochrome AP N-demethylation was considerably stimulated. After primidone/phenobarbitone, especially in combination with phenytoin, /sup 13/C elimination reaches and even surpasses the range for older children. Valproate exposure during fetal life is not consistently followed by a significant increase in AP N-demethylation. The enzyme induction demonstrated by /sup 13/C-AP breath ...

1983-01-01

290

Characterization of commercial cellulases and their use in the saccharification of a sugarcane bagasse sample pretreated with dilute sulfuric acid.

The affinity label 8-((4-bromo-2,3-dioxobutyl)thio)adenosine 5{prime}-triphosphate (8-BDB-TA-5{prime}-TP) reacts covalently with rabbit muscle pyruvate kinase, incorporating 2 mol of reagent/mol of enzyme subunit upon complete inactivation. Protection against inactivation is provided by phosphoenolpyruvate, K{sup +}, and Mn{sup 2+} and only 1 mol of reagent/mol of subunit is incorporated. The authors have now identified the resultant modified residues. After reaction with 8-BDB-TA-5{prime}-TP at pH 7.0, modified enzyme was incubated with ({sup 3}H)NaBH{sub 4} to reduce the carbonyl groups of enzyme-bound 8-BDB-TA-5{prime}-TP and to introduce a radioactive tracer into the modified residues. Following carboxymethylation and digestion with trypsin, the radioactive peptides were separated on a phenylboronate agarose column followed by reverse-phase high-performance liquid chromatography in 0.1% trifluoroacetic acid with an ...

1990-03-13

291

[In vitro digestibility of a milk substitute produced with uncooked flour].

This study aimed to correlate the efficiency of enzymatic hydrolysis of the cellulose contained in a sugarcane bagasse sample pretreated with dilute H(2)SO(4) with the levels of independent variables such as initial content of solids and loadings of enzymes and surfactant (Tween 20), for two cellulolytic commercial preparations. The preparations, designated cellulase I and cellulase II, were characterized regarding the activities of total cellulases, endoglucanase, cellobiohydrolase, cellobiase, ?-glucosidase, xylanase, and phenoloxidases (laccase, manganese and lignin peroxidases), as well as protein contents. Both extracts showed complete cellulolytic complexes and considerable activities of xylanases, without activities of phenoloxidases. For the enzymatic hydrolyses, two 2(3) central composite full factorial designs were employed to evaluate the effects caused by the initial content of solids (1.19-4.81%, w/w) and loadings of enzymes ...

2010-10-17

292

Effect of Lactobacillus supplementation with and without arginine on liver damage and bacterial translocation in an acute liver injury model in the rat.

Milk substitutes are used for the breakfast of the recipients of the National School Feeding Programme in Chile. The major ingredients of milk substitutes are milk, sugar and cereal flours. The technical specifications to produce this type of food state, beside their proportions, that the flours must be precooked. This requisite is controlled by determining the gelatinization degree which must be 92% as a minimum. At present, a cheaper new alternative of milk substitute fabricated with uncooked flours and containing an enzymatic preparation has been presented to the Programme. It is postulated that when reconstituted in water under the manufacturer's directions (85 degrees C, 5 min settling time) precooking of the flours is reached. The objective of this work was to evaluate the in vitro digestibility, the effect of the enzymatic preparation (molecular size of starch and dextrose equivalent) and the degree of gelatinization of this product (MSE) when reconstituted under different ...

1996-12-01

293

Vacuolar/Extravacuolar Distribution of Aminopeptidases in Giant Alga Chara australis and Partial Purification of One Such Enzyme 1

In acute liver failure following hepatitis, toxic insults, or after major liver surgery, there is an increased bacterial translocation from the gut. This may explain some of the infectious complications seen in these conditions. To elucidate mechanisms and find possible preventive measures, we investigated the effect of rectal administration of arginine and probiotic bacteria (Lactobacillus spp.) on bacterial translocation and the extent of liver failure. Sprague-Dawley rats were used and five different Lactobacillus strains (Lb. reuteri R2LC, Lb. rhamnosus DSM 6594 (= strain 271), Lb. plantarum DSM 9843 (= strain 299v), Lb. fermentum 8704:3 (= strain 245), and Lb. reuteri (= strain 108) were administered rectally once daily for 8 days with and without 2% arginine. Acute liver injury (ALI) was induced on the eighth day by intraperitoneal injection of D-galactosamine (1.1 g/kg body weight), and samples were collected after 24 and 48 hours. Bacterial translocation was evaluated by ...

1997-03-01

294

Tungsten Effects on Phosphate-Dependent Biochemical Pathways are Species and Liver Cell Line Dependent

The presence of two major aminopeptidases (aminopeptidases I and II) in the giant alga Chara australis was shown using polyacrylamide gel electrophoresis. Partially purified aminopeptidase I had a molecular weight of about 120,000, hydrolyzed both leucine-?-naphthylamide (pH optimum 6.0) and alanine-?-naphthylamide (pH optimum 7.5), and was located both inside and outside the vacuole. Aminopeptidase I was inhibited by p-chloromercuribenzoic acid, iodoacetic acid, 1,10-phenanthroline, and N-tosyl-l-phenylalanine chloromethyl ketone. Aminopeptidase II hydrolyzed alanine-?-naphthylamide but not leucine-?-naphthylamide and was located only outside the vacuole.ImagesFig. 1Fig. 2Fig. 4Fig. 5

1987-01-01

295

British Library Electronic Table of Contents (United Kingdom)

Tungsten, in the form of tungstate, polymerizes with phosphate, and as extensive polymerization occurs, cellular phosphorylation and dephosphorylation reactions may be disrupted, resulting in negative effects on cellular functions. A series of studies were conducted to evaluate the effect of tungsten on several phosphate-dependent intracellular functions, including energy cycling (ATP), regulation of enzyme activity (cytosolic protein tyrosine kinase [cytPTK] and tyrosine phosphatase), and intracellular secondary messengers (cyclic adenosine monophosphate [cAMP]). Rat noncancerous hepatocyte (Clone-9), rat cancerous hepatocyte (H4IIE), and human cancerous hepatocyte (HepG2) cells were exposed to 1-1000 mg/l tungsten (in the form of sodium tungstate) for 24 h, lysed, and analyzed for the ab...

2010-01-01

296

Transformation by complementation of an adenine auxotroph of the lignin-degrading basidiomycete Phanerochaete chrysosporium

Thyroid Cathepsin K: Roles in Physiology and Thyroid Disease

Swollen basiodiospores of an adenine auxotroph of Phanerochaete chrysosporium were protoplasted with Novozyme 234 and transformed to prototrophy by using a plasmid containing the gene for an adenine biosynthetic enzyme from Schizophyllum commune. Transformation frequencies of 100 transformants per {mu}g of DNA were obtained. Southern blot analysis of DNA extracted from transformants demonstrated that plasmid DNA was integrated into the chromosomal DNA in multiple tandem copies. Analysis of conidia and basiodiospores from transformants demonstrated that the transforming character was mitotically and meiotically stable on both selective and nonselective media. Genetic crosses between double mutants transformed for adenine prototrophy and other auxotrophic strains yielded Ade{sup {minus}} progeny, which indicated that integration occurred at a site(s) other than the resident adenine biosynthetic gene.

1989-02-01

297

British Library Electronic Table of Contents (United Kingdom)

The human genome encodes 11 cysteine cathepsins belonging to the papain-like family of cysteine peptidases that are known predominantly as endo-lysosomal enzymes. However, it is now understood that the functions and activities of cysteine cathepsins are not limited to endo-lysosomal compartments, as they are also active in the peri- and extracellular space. The thyroid gland is an endocrine organ where such intra- and extracellular proteolytic activities are required to solubilize the prohormone thyroglobulin from its luminal, covalently cross-linked storage forms for subsequent processing into smaller protein fragments and thyroid hormone liberation. Cathepsin K has been identified as one of the cysteine cathepsins with a crucial role in thyroglobulin processing. However, cathepsin K has ...

2011-01-01

298

The disruption of l-carnitine metabolism by aluminum toxicity and oxidative stress promotes dyslipidemia in human astrocytic and hepatic cells

British Library Electronic Table of Contents (United Kingdom)

l-Carnitine is a critical metabolite indispensable for the metabolism of lipids as it facilitates fatty acid transport into the mitochondrion where b-oxidation occurs. Human astrocytes (CCF-STTG1 cells) and hepatocytes (HepG2 cells) exposed to aluminum (Al) and hydrogen peroxide (H2O2), were characterized with lower levels of l-carnitine, diminished b-oxidation, and increased lipid accumulation compared to the controls. g-Butyrobetainealdehyde dehydrogenase (BADH) and butyrobetaine dioxygenase (BBDOX), two key enzymes mediating the biogenesis of l-carnitine, were sharply reduced during Al and H2O2 challenge. Exposure of the Al and H2O2-treated cells to a-ketoglutarate (KG), led to the recovery of l-carnitine production with the concomitant reduction in ROS levels. It appears that the chann...

2011-01-01

299

Specificity of the human proteoglycan radioimmunoassay

Species and Organ Diversity in the Effects of Hydrogen Peroxide on Superoxide Dismutase Activity In Vitro

The human articular cartilagineous proteoglycans (PG) R.I.A. is highly specific. The PG used as the standard and the /sup 125/I labelled molecule appear to be pure. Under these conditions, all the potential interfering substances which have been tested show no cross reaction. For instance, the Ag-Ab equilibrium is not affected by adding human IgG, human albumin, hyaluronic acid, chondroitin sulfate, rat type II collagen or total human serum proteins. This R.I.A. also exhibits a species spcificity since there is no cross reaction with rat PG and negligible cross section with dog PG. The results obtained after addition of enzymes to the antigen demonstrate that the antigenic sites are localized on the protein region and not on the glycosaminoglycan region of the molecule.

1981-01-01

300

British Library Electronic Table of Contents (United Kingdom)

Abstract Superoxide dismutase (SOD) is ubiquitous in aerobic organisms and constitutes the first link in the enzyme scavenging system of reactive oxygen species. In the present study, species and organ diversity of SOD activity in a solution and in an in-gel assay system, as well as the effects of hydrogen peroxide (H2O2) on SOD activity, were investigated. In a solution assay system, SOD activity of jackfruit root, shoot, leaves, axes, and cotyledons, of maize embryos and endosperms, of mung bean leaves and seeds, of sacred lotus axes and cotyledons, and of rice and wheat leaves was increased by 1-15 mmol/L H2 O2. However, SOD activity in rice root and seeds, maize roots and leaves, mung bean roots and shoots, and wheat seeds was decreased by 1-15 mmol/L H2O2. The SOD activity of wheat ro...

2006-01-01

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301

Sorbent extraction and high performance liquid chromatography (HPLC) of homidium bromide and isometamidium chloride in bovine plasma

British Library Electronic Table of Contents (United Kingdom)

Homidium bromide and isometamidium chloride are used extensively in the treatment of bovine trypanosomiasis in Africa, but no specific and sensitive method to detect the drug in plasma has been published yet. Comparing the methods of other investigators it was found that they could not overcome the protein binding of these drugs which reduced the available and determined drug to 10% of the real concentrations. These techniques overcome the protein binding by enzyme digestion and an alteration of the pH before adding the sample directly on clean-up columns. Drug recover rates above 80% were obtained. For the detection, the HPLC was employed using a C18 reversed phase analytical column and UV detection, determining both drugs as intact molecules. (author).

1992-01-01

302

Abstract Capillary permeability is a tightly regulated feature of microcirculation in all organ beds. In sepsis, this feature is fundamentally altered. We have previously reported elevated levels of angiopoietin-2 in patients with septic shock, and have investigated tumor necrosis factor (TNF)-related and weak inducer of apoptosis (TWEAK), which mediates both angiogenesis and inflammation, in those patients. Enzyme-linked immunoassay was used to measure serum TWEAK levels in 20 patients with septic shock, all of whom were treated by direct hemoperfusion with a polymyxin B-immobilized fiber column (DHP-PMX), and in 20 non-septic controls. The TWEAK levels were higher in patients with septic shock (192.8--230.5-pg/mL) than in controls (84.1--28.7-pg/mL, P-=-0.043). Between 11 survivors and 1...

2011-01-01

303

Sequential injection system for phospholipase A2 activity evaluation: Studies on liposomes using an environment-sensitive fluorescent probe

British Library Electronic Table of Contents (United Kingdom)

This work reports the development of an automatic methodology based on the use of 1-anilinonaphthalene-8-sulfonate (ANS) as an interfacial fluorescent probe for detecting the hydrophobic environment shift around the probe, caused by the hydrolytic action of PLA2 on the liposomes. The implementation of this reaction in a sequential injection analysis (SIA) system along with the use of the mixing chambers permitted the evaluation of PLA2 activity and assessment of the inhibitory effect of the non-steroidal anti-inflammatory drugs (NSAIDs) on PLA2 activity. Several studies were performed with the aim of establishing the appropriate flow system configuration: the liposome substrate; PLA2 and ANS optimum concentrations and incubation times before and after the enzyme addition. Based on these st...

2009-01-01

304

Response to measles-mumps-rubella vaccine in children on dialysis.

Regulation of C-type natriuretic peptide expression

Ten children receiving maintenance dialysis were immunized with the standard dose of measles-mumps-rubella vaccine between 15 and 33 months of age. Immune responses to vaccination were determined using commercially available enzyme-linked immunosorbent assays for measles, mumps, and rubella viruses. Eight children responded to measles vaccine, 5 to mumps vaccine, 8 to rubella vaccine, and only 3 children to all three vaccines, compared with a seroconversion rate of over 90% to all three vaccines in healthy children (P less than 0.0001). We contend that the relatively poor immunocompetence of our dialysis patients explains their less than optimal vaccine response and suggest that children vaccinated while undergoing dialysis be tested to confirm serological evidence of immunity. PMID:1571219

1992-03-01

305

British Library Electronic Table of Contents (United Kingdom)

C-type natriuretic peptide (CNP) is a member of the small family of natriuretic peptides that also includes atrial natriuretic peptide (ANP) and brain, or B-type natriuretic peptide (BNP). Unlike them, it performs its major functions in an autocrine or paracrine manner. Those functions, mediated through binding to the membrane guanylyl cyclase natriuretic peptide receptor B (NPR-B), or by signaling through the non-enzyme natriuretic peptide receptor C (NPR-C), include the regulation of endochondral ossification, reproduction, nervous system development, and the maintenance of cardiovascular health. To date, the regulation of CNP gene expression has not received the attention that has been paid to regulation of the ANP and BNP genes. CNP expression in vitro is regulated by TGF-b and recepto...

2011-01-01

306

Regulation and secretion of Xanthomonas virulence factors

British Library Electronic Table of Contents (United Kingdom)

Abstract Plant pathogenic bacteria of the genus Xanthomonas cause a variety of diseases in economically important monocotyledonous and dicotyledonous crop plants worldwide. Successful infection and bacterial multiplication in the host tissue often depend on the virulence factors secreted including adhesins, polysaccharides, LPS and degradative enzymes. One of the key pathogenicity factors is the type III secretion system, which injects effector proteins into the host cell cytosol to manipulate plant cellular processes such as basal defense to the benefit of the pathogen. The coordinated expression of bacterial virulence factors is orchestrated by quorum-sensing pathways, multiple two-component systems and transcriptional regulators such as Clp, Zur, FhrR, HrpX and HpaR. Furthermore, virule...

2010-01-01

307

Recent research efforts in the area of biotechnology for fuels and chemicals: Poster session papers

Ramiprilate Inhibits Functional Matrix Metalloproteinase Activity in Crohn-s Disease Fistulas

This report presents research presented at the poster session of the Symposium covering a wide spectrum of current biotechnological research activities. Research focused mostly on ethanol production and methane generation from biomass material via microbial processing, as well as on enhanced hydrogen yield from algae. Several of the posters dealt with the pretreatment of cellulosic materials, and enzyme production/characterization, while a good number of papers displayed research efforts on bioremediation, photosynthesis, production of various useful chemicals from biomass by bioprocessing, and on other miscellaneous subjects. One of the papers treated a very interesting topic of cellulose-cellulase complexes. Many of the poster papers are included in this volume, and a synopsis of all the poster/papers presented is the subject of this article.

1992-01-01

308

British Library Electronic Table of Contents (United Kingdom)

Abstract:- Increased expression of matrix metalloproteinase (MMP)-2, -3 and -9 has been demonstrated in Crohn-s disease fistulas, but it is unknown whether these enzymes are biologically active and represent a therapeutic target. Therefore, we investigated the proteolytic activity of MMPs in fistula tissue and examined the effect of inhibitors, including clinically available drugs that beside their main action also suppress MMPs. Fistula specimens were obtained by surgical excision from 22 patients with Crohn-s disease and from 10 patients with fistulas resulting from other causes. Colonic endoscopic biopsies from six controls were also included. Total functional MMP activity was measured by a high-pressure liquid chromatography (HPLC)-based, fluorogenic MMP-substrate cleavage assay, and t...

2011-01-01

309

Protective effects of long term dietary restriction on swimming exercise-induced oxidative stress in the liver, heart and kidney of rat

British Library Electronic Table of Contents (United Kingdom)

In this study, we evaluated the hypothesis that long term dietary restriction would have beneficial effects on the oxidative stress and antioxidant enzyme systems in liver, heart and kidney in adult male rats undergoing different intensities of swimming exercise. Sixty male, Sprague-Dawley rats were assigned as either dietary restricted on every other week day (DR) or fed ad libitum (AL) groups, and each group was further subdivided into sedentary, endurance swimming exercise training (submaximal exercise) and exhaustive swimming exercise (maximal exercise) groups. Animals in the submaximal exercise group swam 5 days/week for 8 weeks, while maximal exercise was performed as an acute bout of exercise. In parallel with the increase in the intensity of the exercise, the degree of lipid peroxi...

2007-01-01

310

Post harvest improvement of zeaxanthin content of vegetables

British Library Electronic Table of Contents (United Kingdom)

Zeaxanthin is a carotenoid produced by plants and has been associated with protection of the photosynthetic machinery under light stress and, together with lutein, in protection of the central retina of the eye. Zeaxanthin levels in blood plasma have been negatively correlated to the development of AMD (age-related macular degeneration) (Gale et al., 2003). Under normal conditions, plants have a low content of zeaxanthin. The aim of this study was to increase the zeaxanthin content in green vegetables by post harvest treatments. Efficient conditions for activation of the endogenous enzyme system generating zeaxanthin was established and included incubation at low pH (2.5-5.5), with the membrane permeable acetic acid/acetate buffer at room temperature or above for 30min or more. Typically m...

2010-01-01

311

Physiological and antioxidant responses of Mentha pulegium (Pennyroyal) to salt stress

British Library Electronic Table of Contents (United Kingdom)

Mentha pulegium L. is a medicinal and aromatic plant belonging to the Labiatae family present in the humid to the arid bioclimatic regions of Tunisia. We studied the effect of different salt concentrations on plant growth, mineral composition and antioxidant responses. Physiological and biochemical parameters were assessed in the plant organs after 2?weeks of salt treatment with 25, 50, 75 and 100?mM NaCl. Results showed that, growth was reduced even by 25?mM, and salt effect was more pronounced in shoots (leaves and stems) than in roots. This growth decrease was accompanied by a restriction in tissue hydration and K+ uptake, as well as an increase in Na+ levels in all organs. Considering the response of antioxidant enzymes to salt, leaves and roots reacted differently to saline conditions...

2010-01-01

312

Phorbol esters, protein kinase C, and thyroxine 5 prime -deiodinase in brown adipocytes

Pectinolytic yeast isolates for cold-active polygalacturonase production

Protein kinase C activity has been identified in the rat brown adipocyte. About 60% of this activity is found in the cytosolic fraction under basal conditions, and 12-O-tetradecanoylphorbol 13-acetate (TPA) causes a rapid shift from the cytosol to the particulate fraction. Norepinephrine and phenylephrine causes a similar redistribution that can be blocked by prazosin but not by alprenolol. {alpha}{sub 1}-Adrenergic agonists cause three- to fivefold stimulation of type 2 iodothyronine 5{prime}-deiodinase activity in brown adipocytes. TPA has no effect on basal deiodinase activity and reduces the response of the enzyme to {alpha}{sub 1}-adrenergic agonists. These results suggest that the translocation of protein kinase C from cytosol to particulate fraction is not sufficient to increase deiodinase activity but can modulate the {alpha}{sub 1}-adrenergic agonist-mediated responses in these cells.

1988-03-01

313

British Library Electronic Table of Contents (United Kingdom)

Pectin rich cold stored spoiled fruits, vegetables and cold soils were screened and different pectinolytic isolates were obtained by enrichment culturing and ruthenium red plate assay. Among the primary isolates 10-15% were yeast isolates. Six isolates with higher zones of pectin hydrolysis were selected and tested for polygalacturonase (PGU) production at room temperature (25 degrees C) and at 5 degrees C. One isolate identified as Saccharomyces sp. with highest polygalacturonase activity at 5 degrees C was used for enzyme production using raw fruit pectins as substrates. The isolate was identified by preliminary cultural, morphological and sugar fermentation tests. PGU production was high in raw pectin substrates like orange peel (21 U/ml), apple peel (20 U/ml ), mango peel (19 U/ml), ...

2011-01-01

314

Novel Techniques and Their Wide Applications to Health Foods, Medical and Agricultural Biotechnology in Relation to Policy Making on Genetically Modified Crops and Foods

CERN Document Server

Selected applications of novel techniques in Agricultural Biotechnology, Health Food formulations and Medical Biotechnology are being reviewed with the aim of unraveling future developments and policy changes that are likely to open new markets for Biotechnology and prevent the shrinking or closing of existing ones. Amongst the selected novel techniques with applications in both Agricultural and Medical Biotechnology are: immobilized bacterial cells and enzymes, microencapsulation and liposome production, genetic manipulation of microorganisms, development of novel vaccines from plants, epigenomics of mammalian cells and organisms, and biocomputational tools for molecular modeling related to disease and Bioinformatics. Both fundamental and applied aspects of the emerging new techniques are being discussed in relation to their anticipated, marked impact on future markets and present policy changes that are needed for success in either Agricultural or Medical ...

2004-01-01

315

Myocardial pharmacokinetics of ebastine, a substrate for cytochrome P450 2J, in rat isolated heart

British Library Electronic Table of Contents (United Kingdom)

BACKGROUND AND PURPOSE It is well established that cytochrome P450 2J (CYP2J) enzymes are expressed preferentially in the heart, and that ebastine is a substrate for CYP2J, but it is not known whether ebastine is metabolized in myocardium. Therefore, we investigated its pharmacokinetics in the rat isolated perfused heart. EXPERIMENTAL APPROACH Rat isolated hearts were perfused in the recirculating mode with ebastine for 130-min. The concentrations of ebastine and its metabolites, hydroxyebastine and carebastine, were measured using liquid chromatography with a tandem mass spectrometry. The data were analysed by a compartmental model. The time course of negative inotropic response was linked to ebastine concentration to determine the concentration-effect relationship. KEY RESULTS Ebastine w...

2011-01-01

316

Mitochondrial NADH:ubiquinone oxidoreductase (complex I) in eukaryotes: A highly conserved subunit composition highlighted by mining of protein databases

British Library Electronic Table of Contents (United Kingdom)

Complex I (NADH:ubiquinone oxidoreductase) is the largest enzyme of the mitochondrial respiratory chain. Compared to its bacterial counterpart which encompasses 14-17 subunits, mitochondrial complex I has almost tripled its subunit composition during evolution of eukaryotes, by recruitment of so-called accessory subunits, part of them being specific to distinct evolutionary lineages. The increasing availability of numerous broadly sampled eukaryotic genomes now enables the reconstruction of the evolutionary history of this large protein complex. Here, a combination of profile-based sequence comparisons and basic structural properties analyses at the protein level enabled to pinpoint homology relationships between complex I subunits from fungi, mammals or green plants, previously identified...

2011-01-01

317

Metabolic and molecular stress responses of gilthead seam bream Sparus aurata during exposure to low ambient temperature: an analysis of mechanisms underlying the winter syndrome

British Library Electronic Table of Contents (United Kingdom)

The winter syndrome in the gilthead sea bream Sparus aurata indicates that the species is exposed to critically low temperatures in Mediterranean aquaculture in winter. The present study of metabolic patterns and molecular stress responses during cold exposure was carried out to investigate this ?disease?, in light of the recent concept of oxygen and capacity limited thermal tolerance. The metabolic profile of fuel oxidation was examined by determining the activities of the enzymes hexokinase (HK), aldolase (Ald), pyruvate kinase (PK), l-lactate dehydrogenase (l-LDH), citrate synthase (CS), malate dehydrogenase (MDH) and 3-hydroxyacyl CoA dehydrogenase (HOAD) in heart, red and white muscle after exposure to temperatures of 10, 14 and 18?C. Especially, the increase in LDH activity combined ...

2010-01-01

318

Mechanical wounding induces a nitrosative stress by down-regulation of GSNO reductase and an increase in S-nitrosothiols in sunflower (Helianthus annuus) seedlings

British Library Electronic Table of Contents (United Kingdom)

Nitric oxide (NO) and related molecules such as peroxynitrite, S-nitrosoglutathione (GSNO), and nitrotyrosine, among others, are involved in physiological processes as well in the mechanisms of response to stress conditions. In sunflower seedlings exposed to five different adverse environmental conditions (low temperature, mechanical wounding, high light intensity, continuous light, and continuous darkness), key components of the metabolism of reactive nitrogen species (RNS) and reactive oxygen species (ROS), including the enzyme activities L-arginine-dependent nitric oxide synthase (NOS), S-nitrosogluthathione reductase (GSNOR), nitrate reductase (NR), catalase, and superoxide dismutase, the content of lipid hydroperoxide, hydrogen peroxide, S-nitrosothiols (SNOs), the cellular level of N...

2011-01-01

319

Long-distance transport, vacuolar sequestration, tolerance, and transcriptional responses induced by cadmium and arsenic

British Library Electronic Table of Contents (United Kingdom)

Iron, zinc, copper and manganese are essential metals for cellular enzyme functions while cadmium, mercury and the metalloid arsenic lack any biological function. Both, essential metals, at high concentrations, and non-essential metals and metalloids are extremely reactive and toxic. Therefore, plants have acquired specialized mechanisms to sense, transport and maintain essential metals within physiological concentrations and to detoxify non-essential metals and metalloids. This review focuses on the recent identification of transporters that sequester cadmium and arsenic in vacuoles and the mechanisms mediating the partitioning of these metal(loid)s between roots and shoots. We further discuss recent models of phloem-mediated long-distance transport, seed accumulation of Cd and As and rec...

2011-01-01

320

Leishmania major ascorbate peroxidase overexpression protects cells against reactive oxygen species-mediated cardiolipin oxidation

British Library Electronic Table of Contents (United Kingdom)

Heme peroxidases are a class of multifunctional redox-active proteins found in all organisms. We recently cloned, expressed, and characterized an ascorbate peroxidase from Leishmania major (LmAPX) that was capable of detoxifying hydrogen peroxide. Localization studies using green fluorescent protein fusions revealed that LmAPX was localized within the mitochondria by its N-terminal signal sequence. Subcellular fractionation analysis of the cell homogenate by the Percoll density-gradient method and subsequent Western blot analysis with anti-LmAPX antibody further confirmed the mitochondrial localization of mature LmAPX. Submitochondrial fractionation analysis showed that the mature enzyme (?3.6?kDa shorter than the theoretical value of the whole gene) was present in the intermembrane space ...

2008-01-01

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321

Ionizing radiation is a potent inducer of mitotic recombination in mouse embryonic stem cells

British Library Electronic Table of Contents (United Kingdom)

Maintenance of genomic integrity in embryonic cells is pivotal to proper embryogenesis, organogenesis and to the continuity of species. Cultured mouse embryonic stem cells (mESCs), a model for early embryonic cells, differ from cultured somatic cells in their capacity to remodel chromatin, in their repertoire of DNA repair enzymes, and in the regulation of cell cycle checkpoints. Using 129XC3HF1 mESCs heterozygous for Aprt, we characterized loss of Aprt heterozygosity after exposure to ionizing radiation. We report here that the frequency of loss of heterozygosity mutants in mESCs can be induced several hundred-fold by exposure to 5-10Gy of X-rays. This induction is 50-100-fold higher than the induction reported for mouse adult or embryonic fibroblasts. The primary mechanism underlying the...

2011-01-01

322

Impact of different cultivation and induction regimes on the structure of cytosolic inclusion bodies of TEM1-b-lactamase

British Library Electronic Table of Contents (United Kingdom)

The enzyme TEM1-b-lactamase has been used as a model to study the impact of different cultivation and induction regimes on the structure of cytosolic inclusion bodies (IBs). The protein has been heterologously expressed in Escherichia coli in fed-batch cultivations at different temperatures (30, 37, and 40degreeC) as well as induction regimes that guaranteed distinct product formation rates and ratios of soluble to aggregated protein. Additionally, shake flask cultivations at 20, 30, and 37degreeC were performed. IBs were sampled during the whole bioprocess and structural analysis was performed by attenuated total reflectance Fourier transform infrared (ATR-FT-IR) spectroscopy. This work clearly demonstrates that the tested production regimes and rates had no impact on the IB structure, wh...

2008-01-01

323

Immobilization of bacteria in microgel grafted onto macroporous polyethylene

Identification of mutations leading to the Lesch-Nyhan syndrome by automated direct DNA sequencing of in vitro amplified cDNA

The development of 'Green Chemistry' requires new materials to replace the conventional organic chemistry by biological catalysts, to produce fine chemicals in an environmentally friendly manner. Microbial whole cells can be directly used as biocatalysts, providing a simple and cheap methodology since enzyme isolation and purification are avoided. High-density polyethylene (HDPE) is a very stable polymer though it can be activated by gamma radiation to induce grafting. Glycidyl methacrylate was grafted onto macroporous HDPE and PP in the range of 1-6%, proportional to the initial monomer concentration. Grafted polymers were further chemically modified with ethylenediamine to generate a cationic hydrogel of micron-size thickness onto the internal polymer surfaces. Modified polymers were able to immobilize Gram-positive and Gram-negative bacteria that can catalyze a chemical reaction as efficient as free cells do.

2010-03-15

324

How microcystin-degrading bacteria express microcystin degradation activity

The Lesch-Nyhan (LN) syndrome is a severe X chromosome-linked disease that results from a deficiency of the purine salvage enzyme hypoxanthine phosphoribosyltransferase (HPRT). The mutations leading to the disease are heterogeneous and frequently arise as de novo events. The authors have identified nucleotide alterations in 15 independently arising HPRT-deficiency cases by direct DNA sequencing of in vitro amplified HPRT cDNA. They also demonstrate that the direct DNA sequence analysis can be automated, further simplifying the detection of new mutations at this locus. The mutations include DNA base substitutions, small DNA deletions, a single DNA base insertion, and errors in RNA splicing. The application of these procedures allows DNA diagnosis and carrier identification by the direct detection of the mutant alleles within individual families affected by LN.

1989-03-01

325

British Library Electronic Table of Contents (United Kingdom)

Abstract Alkali tolerance and the mechanism of microcystin (MC) degradation were investigated in the MC-degrading bacterial species, Sphingopyxis sp. C-1, to better understand the increased MC degradation under the alkaline conditions that arise during the disappearance of water blooms. MC-degrading bacteria harbour mlrA, mlrB and mlrC that encode MC-degrading enzymes. Sphingopyxis sp. C-1 also possesses these genes, as well as the mlrD gene that has been assumed to encode MC and its degradation transporter. This study demonstrated that MC degradation activity was promoted by the intermittent addition of microcystin-LR (MCLR) to cultures of strain C-1. That the expression of mlrA, mlrB and mlrC is induced by MCLR also was indicated, whereas that of mlrA and mlrB is induced by the MCLR degr...

2011-01-01

326

Green Tea Extract (Epigallocatechin-3-Gallate) Reduces Efficacy of Radiotherapy on Prostate Cancer Cells

British Library Electronic Table of Contents (United Kingdom)

ObjectivesTo assess the influence of epigallocatechin-3-gallate (EGCG) on the efficacy of ionizing radiation on prostate cancer cells because of the increased use of dietary interventions, especially by patients with prostate cancer. Radiotherapy is used to treat localized prostate cancer. Some people consume green tea (EGCG) as a chemopreventive agent against prostate cancer. Green tea can act as an antioxidant and induce superoxide dismutase enzymes, which could scavenge the free oxygen radicals generated by radiotherapy. MethodsProstate cancer cell line DU145 cells were treated with EGCG or radiotherapy, or both. Cell death was assessed using trypan blue cell counting, and apoptosis was confirmed by assessing poly (adenosine phosphate ribose) polymerase cleavage. The antioxidant potenti...

2011-01-01

327

Glutathione peroxidase activity in the selenium-treated alga Scenedesmus quadricauda

British Library Electronic Table of Contents (United Kingdom)

The function of selenium in an organism is mediated mostly by selenoproteins including glutathione peroxidase. Glutathione peroxidase is a potent anti-oxidative enzyme, scavenging a variety of peroxides. The green alga Scenedesmus quadricauda was used to investigate the relationship between the toxicity of selenium and the glutathione peroxidase activity. Selenium resistant strains SeIV and SeVI were synchronized and grown in high concentrations of Se (selenite or selenate). As a measure of selenium toxicity the EC50 values were determined. During growth of the untreated wild type, glutathione peroxidase activity increased slightly and then declined gradually until the end of the cell cycle. A similar pattern was observed in untreated resistant strains and when resistant strains were grown...

2011-01-01

328

Genetic structure of gilthead seabream, Sparus aurata, in the Central Mediterranean Sea

British Library Electronic Table of Contents (United Kingdom)

The gilthead seabream, Sparus aurata, represents an important economic resource for Mediterranean aquaculture. In spite of its wide geographic distribution and economic importance, only recently studies have been carried out on the genetic composition of natural populations, which have revealed a picture of a heterogeneous degree of genetic differentiation among S. aurata populations. In this study an allozyme analysis of samples from six different collecting sites along the Italian and Croatian coasts was carried out, covering an area in the Central Mediterranean sea that has yet to be investigated through gene-enzyme systems. Data on 26 gene loci, 10 of which are polymorphic, indicate a slight but significant genetic structure (FST = 0.0167) of the species. A hierarchical analysis of pop...

2006-01-01

329

Food-grade gene expression in lactic acid bacteria

British Library Electronic Table of Contents (United Kingdom)

Abstract In the 1990s, significant efforts were invested in the research and development of food-grade expression systems in lactic acid bacteria (LAB). At this time, Lactococcus lactis in particular was demonstrated to be an ideal cell factory for the food-grade production of recombinant proteins. Steady progress has since been made in research on LAB, including Lactococcus, Lactobacillus and Streptococcus, in the areas of recombinant enzyme production, industrial food fermentation, and gene and metabolic pathway regulation. Over the past decade, this work has also led to new approaches on chromosomal integration vectors and host/vector systems. These newly constructed food-grade gene expression systems were designed with specific attention to self-cloning strategies, food-grade selection...

2011-01-01

330

Enzymatic solubilization of a pectinaceous dietary fiber fraction from potato pulp: Optimization of the fiber extraction process

British Library Electronic Table of Contents (United Kingdom)

Upgrading of potato pulp, a byproduct stream from industrial manufacture of potato starch, is important for the continued economic competitiveness of the potato starch industry. The major part of potato pulp consists of the tuber plant cell wall material which is particularly rich in galactan branched rhamnogalacturonan I type pectin. In the work reported here, the release of high-molecular weight pectinaceous dietary fiber polysaccharides from starch free potato pulp was accomplished by use of a multicomponent pectinase preparation from Aspergillus aculeatus (Viscozyme L). The enzyme reaction conditions for the solubilization were optimized via a surface response design to be addition of 0.27% Viscozyme L by weight of potato pulp substrate dry matter, 1h treatment at pH 3.5, 62.5degreeC. ...

2009-01-01

331

Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose; Actividad enzimatica del complejo celulolitico producido por Trichoderma reesei. Hidrolisis enzimatica de la celulosa

Engineered ketol-acid reductoisomerase and alcohol dehydrogenase enable anaerobic 2-methylpropan-1-ol production at theoretical yield in Escherichia coli

The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs.

1986-07-01

332

British Library Electronic Table of Contents (United Kingdom)

2-methylpropan-1-ol (isobutanol) is a leading candidate biofuel for the replacement or supplementation of current fossil fuels. Recent work has demonstrated glucose to isobutanol conversion through a modified amino acid pathway in a recombinant organism. Although anaerobic conditions are required for an economically competitive process, only aerobic isobutanol production has been feasible due to an imbalance in cofactor utilization. Two of the pathway enzymes, ketol-acid reductoisomerase and alcohol dehydrogenase, require nicotinamide dinucleotide phosphate (NADPH); glycolysis, however, produces only nicotinamide dinucleotide (NADH). Here, we compare two solutions to this imbalance problem: (1) over-expression of pyridine nucleotide transhydrogenase PntAB and (2) construction of an NADH-de...

2011-01-01

333

Effects of various maillard reaction products on in vitro starch hydrolysis and blood glucose responses in mice

British Library Electronic Table of Contents (United Kingdom)

Abstract This study investigated the effects of Maillard reaction products (MRPs) on in vitro starch hydrolysis, and in vivo blood glucose responses in mice. Four MRPs of various dextrose equivalents were prepared by heating a mixture of hydrolyzed rice starch and glycine at 200C for 4-min. The starch hydrolysis rates of gelatinized rice starch (GRS) decreased as the browning reaction of added MRPs increased. The hydrolysis kinetic constants of the GRS with 5% MRPs were relatively lower than that of GRS without MRPs. Blood glucose responses showed similar tendencies to the in vitro starch hydrolysis results. The activity of digestive enzymes was inhibited by the MRPs. The relative crystallinity of all MRPs (29.9-60.1%) appeared to have higher values than GRS (15.1%). The samples heated to ...

2011-01-01

334

Effect of season on contractile and metabolic properties of desert camel muscle (Camelus dromedarius)

British Library Electronic Table of Contents (United Kingdom)

Thirty fattened one humped desert camels were used to examine the effect of season on contractile and metabolic properties of Longissimus thoracis (LT) muscle. Ten camels were slaughtered according to seasons of the year (winter, summer and autumn). Season significantly influenced muscle chemical composition, ultimate pH (pHu) and color. Activities of metabolic enzymes were higher during autumn season compared to summer and winter for phosphofructokinase (+64% compared to both seasons) and for isocitrate dehydrogenase (+35% and +145% in autumn vs. summer and winter, respectively). Quantification of muscle myosin heavy chain isoforms by SDS-PAGE electrophoresis showed only presence of type I and type IIa MyHC in camel muscle and indicated high proportion in winter for type I and in autumn f...

2012-01-01

335

Differential expression of farnesyl diphosphate synthase gene from Withania somnifera in different chemotypes and in response to elicitors

British Library Electronic Table of Contents (United Kingdom)

Withania somnifera (L.) Dunal (Family, Solanaceae), commonly known as Ashwagandha is one of the most valuable medicinal plants synthesizing large number of pharmacologically active secondary metabolites known as withanolides. Though the plant has been well characterized in terms of phytochemical profiles as well as pharmaceutical activities, not much is known about the genes responsible for biosynthesis of these compounds. In this study, we have characterized a gene encoding farnesyl diphosphate synthase (FPPS; EC 2.5.1.10), a key enzyme in the pathway of biosynthesis of isoprenoids, from W. somnifera. The full-length cDNA of Withania somnifera FPPS (WsFPPS) of 1,253?bps encodes a polypeptide of 343 amino acids. The amino acid sequence homology and phylogenetic analysis suggest that WsFPPS...

2011-01-01

336

Cyclooxygenase expression and prostaglandin levels in central nervous system tissues during the course of chronic relapsing experimental autoimmune encephalomyelitis (EAE)

British Library Electronic Table of Contents (United Kingdom)

Objective Multiple sclerosis (MS) and its animal counterpart experimental autoimmune encephalomyelitis (EAE) have a major inflammatory component that drives and orchestrates both diseases. One particular group of mediators are the prostaglandins (PGs), which we have previously shown, through quantitation and pharmacological intervention, to be closely involved in the pathology of MS and EAE. The aim of the current study was to determine the expression of the PG-generating cyclooxygenase (COX) enzymes and the profile of PGE2 and PGD2, in selected central nervous system (CNS) tissues, with the development of the chronic relapsing (CR) form of EAE. In particular, the work investigates the possible relationship between the expression of COX isoenzymes and PG levels during the neurological phas...

2011-01-01

337

Cumulative effect of X-ray radiation and inflammatory reaction on the circadian rhythm of tyrosine aminotransferase in the liver of mice

Chromosomal localization and structure of the human type II IMP dehydrogenase gene

The activity of tyrosine aminotransferase (TAT) in the liver of mice subjected simultaneously to ionizing radiation and to local inflammatory process showed in the first 24 hours of the experiment statistically significant changes resulting from summation of the harmful effects of both factors. In the second 24-hour period an evident tendency was observed for return of a normal circadian rhythm of the enzyme. This points to presence of an endogenous mechanism of suppression of the manifestations of significant desynchronization of the circadian rhythm. The paper contains also a trial of mathematical analysis of changes in TAT activity in the circadian rhythm, a description and analysis or the suppression process, qualitative and quantitative determination of the phenomenon of cumulation of the effects of powerful stress factors acting on the animal organism. The obtained results were compared with the results of investigations of TAT activity changes obtained in ...

1980-01-01

338

Biochemical alterations induced by Zn and Cd individually or in combination in the serum of Oreochromis niloticus

We determined the chromosomal localization and structure of the gene encoding human type II inosine 5{prime}-monophosphate dehydrogenase (IMPDH, EC 1.1.1.205), an enzyme associated with cellular proliferation, malignant transformation, and differentiation. Using polymerase chain reaction (PCR) primers specific for type II IMPDH, we screened a panel of human-Chinese hamster cell somatic hybrids and a separate deletion panel of chromosome 3 hybrids and localized the gene to 3p21.1{yields}p24.2. Two overlapping yeast artificial chromosome clones containing the full gene for type II IMPDH were isolated and a physical map of 117 kb of human genomic DNA in this region of chromosome 3 was constructed. The gene for type II IMPDH was localized and oriented on this map and found to span no more than 12.5 kb.

1994-05-01

339

British Library Electronic Table of Contents (United Kingdom)

Measurement of serum biochemical parameters in response to metal exposures can be especially useful to help identify target organs of toxicity as well as the general health status of animals. Oreochromis niloticus were exposed to 5.0?mg/L?Zn, 1.0?mg/L?Cd, and 5.0?mg/L?Zn?+?1.0?mg/L?Cd mixture for 7 and 28?days, and alterations in serum enzyme activities and ion levels were measured. Significant changes in all the biochemical parameters were found to be time dependent. Following metal exposure, alkaline phosphatase activity was elevated at both exposure periods. No changes in activities of lactate dehydrogenase and lipase were observed in response to single or combined Zn and Cd exposure at 7?days while they increased at 28?days. Fish exposed to metals showed a decrease in cholinesterase ac...

2010-01-01

340

Angiopoietin Balance in Septic Shock Patients With Acute Lung Injury: Effect of Direct Hemoperfusion With Polymyxin B-Immobilized Fiber

British Library Electronic Table of Contents (United Kingdom)

Abstract Acute lung injury (ALI) in sepsis is characterized by an increase in microvascular permeability, resulting in pulmonary edema. Several studies have suggested that angiopoietin-1 and -2 play a contributory role in the pathogenesis of ALI. Polymyxin B-immobilized fiber column hemoperfusion is effective for sepsis-induced ALI. We investigated the angiopoietin levels before and after direct hemoperfusion with polymyxin B-immobilized fiber column (PMX) therapy. Enzyme-linked immunoassay was used to measure the serum angiopoietin-1 and -2 levels in 25 patients with septic shock treated with PMX. Eleven of the 25 patients were diagnosed with ALI. There was a significant positive correlation between the angiopoietin-1 level and the PaO2/FiO2 ratio, but there was a significant inverse corr...

2011-01-01

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341

Amelioration of phytotoxic effects of Cd on mung bean seedlings by gluconic acid secreting rhizobacterium Enterobacter asburiae PSI3 and implication of role of organic acid

British Library Electronic Table of Contents (United Kingdom)

Mung bean seedlings inoculated with Enterobacter asburiae PSI3, a gluconic acid-producing rhizosphere isolate, enhanced plant growth in the presence of phytotoxic levels of Cd2+ in gnotobiotic pot experiments as compared to the uninoculated Cd-treated plants. Addition of organic acids to Cd-stressed seedlings promoted root elongation. Hematoxylin competition assays showed that organic acids could displace Cd2+ from the Cd2+: hematoxylin complex in the same order of effectiveness as was found for restoration of root net elongation viz. oxalate?>?malate?>?succinate while gluconate was effective at higher concentrations. Root associated Cd2+, assessed by hematoxylin staining of roots was found to be reduced when roots were treated with organic acid. Cd stress increased antioxidant enzymes suc...

2008-01-01

342

A lytic enzyme cocktail from Streptomyces sp. B578 for the control of lactic and acetic acid bacteria in wine

British Library Electronic Table of Contents (United Kingdom)

Beside yeasts, lactic acid bacteria (LAB) are the most abundant microbes in must during vinification. Whereas Oenococcos oeni is commercially used as a starter culture for the biological acid reduction in wines, other species are responsible for different types of wine spoilage. Members of the genera Pediococcus, Weissella, Leuconostoc, and Lactobacillus are producers of exopolysaccharide slimes, biogenic amines, acetic acid, and other off-flavors. In order to control microbial growth, different procedures such as heating of must and addition of sulfite or lysozyme from egg white are generally applied. Yet, because of health risks, the application of sulfite should be reduced and lysozyme is not effective against all LAB. In this study, we describe exoenzymes from a Streptomyces sp. strain...

2009-01-01

343

[mRNA level and cytochrome P450 1A activity in the liver of C57BL mice induced by various xenobiotics].

[Cloning of the gene for thermostable Thermus aquaticus YT1 DNA polymerase and its expression in Escherichia coli].

The rate of hepatic cytochrome P450 Cypla1 and Cyp1a2 induction was investigated in C57BL male mice during induction with o-aminoazotoluene (OAT), benzo[a]pyrene (BP) and 1,4-dihydroxyanthraquinone (AQ). The Cypla1 mPNA level determined by quantitative RT-competitive PCR increased more than three orders of magnitude during induction with OAT and BP compared with untreated animals and remained unchanged during induction with AQ. The Cypla2 mRNA level was only 8.5, 18.7 and 1.9 times higher during induction with OAT, BP and AQ respectively than in untreated mice. At the same time 7-Ethoxyresorufin-O-deethylase (EROD) and 7-Methoxyresorufin-O-demethylase (MROD) activities of Cypla were also investigated in liver. The increase of Cypla1 mRNA level correlated with the increase of EROD activity. This suggests involvement of the transcriptional mechanism of the inducibility of this enzyme. In the case of Cypla2 there was insignificant increase of its mRNA level but high ...

344

["3H]Indole-3-acetyl-myo-inositol hydrolysis by extracts of Zea mays L. vegetative tissue

Using the phasmid vector pSL5, the genomic DNA fragment of T. aquaticus YT1 which contained the thermostable DNA polymerase (Taq-polymerase) gene was cloned. The BglII fragment of this genome locus was subcloned in the BamHI site of the pUC19 plasmid. To optimize the Taq-polymerase gene expression in E. coli cells, the gene was cloned in the correct reading frame regarding the initiation ATG codon of the pPR-TGATG-1 expression vector. The gene expression in this vector was controlled by the phage lambda PR promoter and the temperature-sensitive phage lambda repressor. We used PCR to amplify the short 5'-end fragment of the Taq-polymerase gene coding for the part into which an artificial SacI site was introduced. This site has been used for cloning the PCR product into the pPR-TGATG-1 vector, and the missing gene part was cloned into the KpnI site of the PCR product from the natural cloned gene. The cells of the E. coli PVG-A1 strain, which was obtained in the end, expressed efficiently ...

345

Zonal differences in DNA synthesis activity and cytochrome P450 gene expression in livers of male F344 rats treated with five nongenotoxic carcinogens

["3H]Indole-3-acetyl-myo-inositol was hydrolyzed by buffered extracts of acetone powders prepared from 4 day shoots of dark grown Zea mays L. seedlings. The hydrolytic activity was proportional to the amount of extract added and was linear for up to 6 hours at 37"0C. Boiled or alcohol denatured extracts were inactive. Analysis of reaction mixtures by high performance liquid chromatography demonstrated that not all isomers of indole-3-acetyl-myo-inositol were hydrolyzed at the same rate. Buffered extracts of acetone powders were prepared from coleoptiles and mesocotyls. The rates of hydrolysis observed with coleoptile extracts were greater than those observed with mesocotyl extracts. Active extracts also catalyzed the hydrolysis of esterase substrates such as #alpha#-naphthyl acetate and the methyl esters of indoleacetic acid and naphthyleneacetic acid. Attempts to purify the indole-3-acetyl-myo-inositol hydrolyzing activity by chromatographic procedures resulted in only slight ...

346

Synthesis of (methine- sup 3 H)DDT and its nitro-analog, and isotope effects in their enzyme-catalyzed dehydrochlorination

Both increased cell proliferation and {open_quotes}altered{close_quotes}CYP gene expression are prominent phenomena associated with liver tumor promotion by nongenotoxic carcinogen treatment. BRDU-labeled parenchymal nuclei were observed primarily in the periportal area of groups of rats, independent of nongenotoxic carcinogen treatment. Treatment with each of the 5 nongenotoxic carcinogens resulted in profound alterations in CPY gene expression at both the transcriptional and translational levels. Expression of CYP1A1, 1A1/2, 3A1, 2B1/2, and 4A immunoproteins demonstrated nongenotoxic carcinogen-specific patterns in both magnitude and zonal distribution. In agreement with the CYP immunoprotein data, treatment with each of the five nongenotoxic carcinogens resulted in a unique composition of mRNAs of CYP2B1, 2B2, 2C6, 2C11, 3A1, 3A2, and 4A1, which were variably increased or decreased relative to the untreated control livers, depending on the treatment. Similarly, the rate and pattern ...

1995-12-31

347

Nuclear magnetic resonance spectroscopy in biomedicine. Course organized by the Istituto Superiore di Sanita`. Marciana Marina (Isola d`Elba), September 18-23, 1995; Spettroscopia di risonanza magnetica nucleare in biomedicina. Corso organizzato dall`Istituto Superiore di Sanita`. Marciana Marina (Isola d`Elba), 18-23 settembre 1995

(methine-{sup 3}H)1,1-Di-(4-chlorophenyl)-2,2,2-trichloroethane ((methine-{sup 3}H)DDT) and its di-(4-nitrophenyl) analog, both of high purity with a moderately high specific activity were prepared. Chloro-benzene was condensed with (1-{sup 3}H)1-(4-chlorophenyl)-2,2,2-trichloro-ethanol, which has been synthesized by sodium boro({sup 3}H)hydride reduction of 4-chlorophenyl trichloromethyl ketone. The purified ({sup 3}H)DDT had a specific activity of 0.77 Ci/mmol (28.49 GBq/mmol). (methine-{sup 3}H)1,1-Diphenyl-2,2,2-trichloroethane was similarly synthesized and was nitrated to give (methine-{sup 3}H)1,1-di-(4-nitrophenyl)-2,2,2-trichloro-ethane of 1.63 Ci/mmol (60.31 GBq/mmol). Dehydrochlorination with housefly enzyme (glutathione-dependent DDT dehydrochlorinase) showed a remarkable isotope effect. For DDT, the observed tritium isotope effect on V{sub max}/K{sub m} was 11.51{plus minus}0.52. For the nitro-analog, the value was 11.3{plus minus}1.2. We measured ...

1989-08-01

348

Liver Zn-thionein (ZnMT) regulates the interaction of Zn and Pb with delta-aminolevulinic acid dehydratase (ALAD)

Nuclear magnetic resonance is a powerful technique that can be used in a wide range of applications, such as the structural characterization of high molecular weight molecules, conformational studies on enzymes in solution, enzyme-substrate or DNA-protein interactions, monitoring of cell metabolism in vivo, and for diagnostic purposes, employing spectroscopic and imaging techniques. This course was organized in order to introduce the participants to the fundamentals of NMR spectroscopy, and offer practical advice on performing NMR experiments on cell systems, cell and tissue extracts and animal models. The main implications regarding human experiments were also discussed. Finally the quantification of information and the interpretation of data were considered with regard to the main nuclei observed. [Italiano] La risonanza magnetica nucleare e` una delle tecniche spettroscopiche che meglio risponde all`ampio spettro di condizioni imposto dalla ...

1997-12-01

349

Involvement of protein kinase C in the mechanism of action of Escherichia coli heat-stable enterotoxin (STa) in a human colonic carcinoma cell line, COLO-205

ZnMT has been postulated to function in essential metal homeostasis and metal detoxication. This study was undertaken to evaluate the potential of liver ZnMT to mediate the interaction of Zn and Pb with the Zn metalloenzyme, ALAD. Pretreatment of rats with Zn activated liver ALAD and attenuated the inhibition of ALAD by Pb in vitro and in vivo. Liver cytosol from Zn-pretreated rats injected with /sup 203/Pb was fractionated via gel filtration and anion-exchange chromatography and disclosed that both Zn and /sup 203/Pb co-eluted in the MT fractions. Both purified ZnMT-I and ZnMT-II bound /sup 203/Pb in vitro as shown by gel filtration chromatography. Addition of purified liver ZnMT to purified bovine liver ALAD reaction mixtures increased enzyme activity by 2-fold and prevented inhibition of ALAD by Pb. Addition of apothionein alone decreased the activity of Zn-activated ALAD and also attenuated Pb inhibition of the enzyme. Gel filtration ...

1987-05-01

350

Insulin regulation of protein biosynthesis in differentiated 3T3 adipocytes. Regulation of glyceraldehyde-3-phosphate dehydrogenase

The present study was undertaken to determine the involvement of calcium-protein kinase C pathway in the mechanism of action of Escherichia coli heat stable enterotoxin (STa) apart from STa-induced activation of guanylate cyclase in human colonic carcinoma cell line COLO-205, which was used as a model cultured cell line to study the mechanism of action of E. coli STa. In response to E. coli STa, protein kinase C (PKC) activity was increased in a time-dependent manner with its physical translocation from cytosol to membrane. Inhibition of the PKC activity in membrane fraction and inhibition of its physical translocation in response to IP_3-mediated calcium release inhibitor dantrolene suggested the involvement of intracellular store depletion in the regulation of PKC activity. Among different PKC isoforms, predominant involvement of calcium-dependent protein kinase C (PKC#alpha#) was specified using isotype-specific pseudosubstrate, which showed pronounce enzyme ...

2005-08-01

351

Induction of biotransformation in the liver of Eel (Anguilla anguilla L. ) by sublethal exposure to dinitro-o-cresol: An ultrastructural and biochemical study

The effect of insulin on protein biosynthesis was examined in differentiated 3T3-L1 and 3T3-F442A adipocytes. Insulin altered the relative rate of synthesis of specific proteins independent of its ability to hasten conversion of the fibroblast (preadipocyte) phenotype to the adipocyte phenotype. Although more than one pattern of response to insulin was observed, the authors focused on the induction of a Mr 33,000 protein which was identified as the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Exposure of 3T3 adipocytes to insulin throughout differentiation specifically increased GAPDH activity and protein content by 2- to 3-fold as compared to 3T3 adipocytes differentiated in the absence of insulin. These changes in enzyme activity and content could be accounted for by a 4-fold increase in the relative rate of synthesis of GAPDH and a 9-fold increase in hybridizable mRNA levels. Within 2 h of insulin addition to 3T3 ...

1985-10-05

352

Expression of bacterial luciferase in eukaryotic cells

Structural and functional alterations in hepatocytes of the European eel, Anguilla anguilla, following a 4-week-exposure to 5, 50, and 250 micrograms/liter dinitro-o-cresol (DNOC) were investigated by means of electron microscopy and biochemistry and compared to liver pathology in eels exposed to the chemical spill into the Rhine river at Basle in November 1986. Whereas phenological parameters (growth, condition factor) are unaffected, ultrastructural and biochemical alterations are detectable at greater than or equal to 50 and 5 micrograms/liter DNOC, respectively. Structural modifications include: rounding-up of the nuclei; fractionation and reduction of the rough endoplasmic reticulum; proliferation of the smooth endoplasmic reticulum (SER), mitochondria, peroxisomes, and lysosomes; bundles of rod-shaped SER profiles; annulate lamellae; membrane whorls within mitochondria; crystallization of the peroxisomal matrix and glycogen bodies; glycogen depletion and lipid augmentation. ...

1991-04-01

353

Crystal Structure of Human Senescence Marker Protein 30: Insights Linking Structural, Enzymatic, and Physiological Functions

Expression of Bacterial luciferase enzyme (lux) in mammalian cells would be a powerful bioreporter protein system for in vivo imaging because eukaryotic luciferases need expensive substrates. However, only a few efforts have been made to express bacterial luciferase enzyme in mammalian cells. As the result of this, we attempted to construct bicistronic vector including two bacterial luciferase genes (LuxA and LuxB) for assessing the potential to be visualized in vitro or in vivo by optical imaging system after transfection to mammalian cells. We designed and synthesized luxA and luxB genes from Photorhabdus Luminescens. To co-express both luxA and luxB genes from a single promoter, we cloned as a bicistronic transcript fused with an internal ribosomal entry site (IRES). This bicistronic transcript was transfected by Superfect to HEK 293T cell line. We also transfected lux A and lux B vector to HEK 293T cells separately. To evaluate gene ...

2005-11-18

354

Atomic substitution reveals the structural basis for substrate adenine recognition and removal by adenine DNA glycosylase

Human senescence marker protein 30 (SMP30), which functions enzymatically as a lactonase, hydrolyzes various carbohydrate lactones. The penultimate step in vitamin-C biosynthesis is catalyzed by this enzyme in nonprimate mammals. It has also been implicated as an organophosphate hydrolase, with the ability to hydrolyze diisopropyl phosphofluoridate and other nerve agents. SMP30 was originally identified as an aging marker protein, whose expression decreased androgen independently in aging cells. SMP30 is also referred to as regucalcin and has been suggested to have functions in calcium homeostasis. The crystal structure of the human enzyme has been solved from X-ray diffraction data collected to a resolution of 1.4 {angstrom}. The protein has a 6-bladed {beta}-propeller fold, and it contains a single metal ion. Crystal structures have been solved with the metal site bound with either a Ca{sup 2+} or a Zn{sup 2+} atom. The catalytic role of the ...

2010-05-25

355

Antiperoxidative and antiinflammatory effect of Sida cordifolia Linn. on quinolinic acid induced neurotoxicity.

Adenine DNA glycosylase catalyzes the glycolytic removal of adenine from the promutagenic A {center_dot} oxoG base pair in DNA. The general features of DNA recognition by an adenine DNA glycosylase, Bacillus stearothermophilus MutY, have previously been revealed via the X-ray structure of a catalytically inactive mutant protein bound to an A:oxoG-containing DNA duplex. Although the structure revealed the substrate adenine to be, as expected, extruded from the DNA helix and inserted into an extrahelical active site pocket on the enzyme, the substrate adenine engaged in no direct contacts with active site residues. This feature was paradoxical, because other glycosylases have been observed to engage their substrates primarily through direct contacts. The lack of direct contacts in the case of MutY suggested that either MutY uses a distinctive logic for substrate recognition or that the X-ray structure had captured a noncatalytically competent state in lesion ...

2010-01-14

356

[Molecular cloning and expression of an isotoxin gene, alpha-bungarotoxin, from Bungarus multicinctus].

Sida cordifolia is a plant belonging to the Malvaceae family used in many ayurvedic preparations. This study aimed at assessing the effects of ethanolic extract of Sida cordifolia root on quinolinic acid (QUIN) induced neurotoxicity and to compare its effect with the standard drug deprenyl in rat brain. Rats were divided into six groups: (1) control group (2) QUIN (55 microg/100 g bwt/day) (3) 50% ethanolic plant extract treated group (50 mg/100 g bwt/day) (4) Deprenyl (100 microg/100 g bwt/day) (5) QUIN (55 microg/100 g bwt/day) + 50% ethanolic plant extract treated group (50 mg/100 g bwt/day) (6) QUIN (55 microg/100 g bwt/day) + Deprenyl (100 microg/100 g bwt/day). At the end of the experimental period a status of lipid peroxidation products, protein peroxidation product, activities of the scavenging enzymes and the activities of the inflammatory markers were analyzed. Results revealed that the lipid peroxidation products decreased and the activities of the ...

2010-05-25

357

[Fe]-hydrogenases in green algae: photo-fermentation and hydrogen evolution under sulfur deprivation

Abstract: Snake venom contains a number of small proteins,enzymes and other components,which displays a broad spectrum of biological activities. With the ability of specifically binding on acetylcholine acceptor, alpha-bungarotoxins are not only useful molecular probes in investigating the mechanism of neural signal transmission, but also potential pharmic preparations for neural disease treatment. In current research,cDNAs of Bungarus multicinutus venom gland were synthesized using SMART cDNA amplification kit and then, alpha-bungarotoxin genes were cloned and sequenced. Total of 20 clones were sequenced representing 14 isotoxin mRNAs of alpha-bungarotoxins. Among those clones, a novel isotoxin gene was subcloned into two expression plasmids, alpha-BgTX/pQE30a and alpha-BgTX/pGEX-4T-1, and transformed into E. coli. After inducing with IPTG, fused protein of GST-alpha-BgTX was successfully expressed at level of 30% gross proteins of bacteria. More than 25% of fused ...

2005-07-01

358

Using light to bioactivate surfaces: A new way of creating oriented, active immunobiosensors

Recent studies indicate that [Fe]-hydrogenases and H{sub 2} metabolism are widely distributed among green algae. The enzymes are simple structured and catalyze H{sub 2} evolution with similar rates than the more complex [Fe]-hydrogenases from bacteria. Different green algal species developed diverse strategies to survive under sulfur deprivation. Chlamydomonas reinhardtii evolves large quantities of hydrogen gas in the absence of sulfur. In a sealed culture of C. reinhardtii, the photosynthetic O{sub 2} evolution rate drops below the rate of respiratory O{sub 2} consumption due to a reversible inhibition of photosystem II, thus leading to an intracellular anaerobiosis. The algal cells survive under these anaerobic conditions by switching their metabolism to a kind of photo-fermentation. Although possessing a functional [Fe]-hydrogenase gene, the cells of Scenedesmus obliquus produce no significant amounts of H{sub 2} under S-depleted conditions. Biochemical ...

2002-12-01

359

The inhibition of mitochondrial DNA polymerase gamma from animal cells by intercalating drugs.

Ultraviolet light can be used to immobilize biomolecules onto thiol reactive surfaces in order to, e.g., make biosensors. The mechanism involves light-induced formation of free, reactive thiol groups in disulphide containing molecules. This technology allows for the creation of arrays of biomolecules with a high degree of reproducibility, circumventing the need for often expensive nano/micro-dispensing technologies. The ultimate size of the immobilized spots is defined by the focal area of the UV beam. Light-induced immobilization has the added benefit that the immobilized molecules will be spatially oriented and covalently bound to the surface. In this paper, we demonstrate the utility of a sensor array created with the new sensor technology when integrated into a microfluidic system. Protein arrays made using light-induced immobilization showed successful antigen/antibody binding in a flow cell allowing the visualisation of real time binding and enzyme activity. ...

2007-12-15

360

Sucrose phosphate synthase activity in rice grown at elevated CO sub 2 and temperature

DNA polymerase gamma from purified nuclei of EMT-6 cells (mice) seems to be identical to the mitochondrial DNA polymerase from the same source following several criteria. These two enzyme activities are strongly inhibited by ethidium bromide and acriflavin, while proflavin, acridine orange, daunomycin and chloroquine inhibition is less pronounced. In the case of DNA polymerases alpha and beta very little inhibition by ethidium bromide was observed. Intercalation of this dye in a poly dA-dT 12-18 template-primer was studied spectrophotometrically under conditions similar to those in the in vitro DNA polymerase assay. The polymerase assay. The inhibition by this drug of the mitochondrial DNA polymerase gamma activity was shown to be competitive at varying concentrations of TTP while the inhibition was of the non-competitive type at different concentrations of poly dA-dT 12-18. We conclude that the drug, most probably in the intercalated form, is able to interact with ...

1978-06-01

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361

Stable isotope studies on iron bio-availability in young Indian women: Effects of nutritional status, interactions and ethnicity

Rice (Oryza sativa L.) was grown at 330 and 660 {mu}L CO{sub 2} L{sup {minus}1} and at 40/33/37, 34/27/31, and 28/21/25{degree}C day/night/paddy water temperatures respectively. Sucrose phosphate synthase (SPS) activity was measured at saturating substrate concentrations at 59 days after planting. SPS activity increased 2 and 3 fold with increasing CO{sub 2} at 28 and 34{degree}C air temperatures respectively. At 40{degree}C SPS activity decreased by 37% at elevated CO{sub 2} and most plants failed to reach maturity. Similar responses were found in leaf samples taken in the dark. These results indicate that SPS, an enzyme involved in the regulation of C partitioning in leaves, increases in activity at elevated CO{sub 2}. This is in contrast to previous results with soybean. The changes in SPS activity will also be discussed in relation to leaf starch/sucrose ratios.

1990-05-01

362

Soil Redox Chemistry Limitation of Selenium Concentration in Carex Species Sedges

Iron is an essential micronutrient. As an integral part of hemoglobin, it is essential for the transport of oxygen in blood to the tissues. It is an important component of cytochromes and other tissue enzymes that are critical for cellular respiration and of myoglobin that helps maintain oxygen reserve in the muscle. It is also involved in the normal functioning of the immune function, and in the synthesis of steroid hormones and bile acids. There are serious functional consequences to iron deficiency and iron deficiency anemia (IDA) in the human body. The deleterious effects include increased morbidity, preterm delivery, low birth weight, delayed cognitive development, lowered cellular immunity and reduced physical work capacity. However, iron deficiency and IDA remain the world's most common deficiency disease in the 21st century. Recent estimates of people affected with iron deficiency and IDA are 3.5 billion. Iron deficiency anemia is most prevalent and severe ...

2002-06-24

363

Selective metabolic stimulation of the subfornical organ and pituitary neural lobe by peripheral angiotensin II

The trace element selenium (Se) is required in the production of enzymes that protect mammalian cells from oxidative damage due to the byproducts of aerobic respiration. Its deficiency in livestock results in the nutritional muscular dystrophy called white muscle disease. This especially affects juveniles in the preweaning period. Symptoms have also been reported in wild herbivores on low-Se forage, and their appearance may be episodic, suggesting temporal variations in Se uptake by plants. Here, we report variations in selenium concentrations in Carex spp. sedges used as forage by bighorn sheep (Ovis canadensis) on summer range in the Wyoming, Wind River Mountains, and correlate those variations with soil conditions that affect the bioavailability of selenium. Variations in available Se are explained based upon the known oxidation/reduction chemistry of the element. It is concluded that water-saturated, alpine soils provide conditions suitable for the reduction of ...

2007-09-01

364

SE and I content in military total diet

The subfornical organ is a major receptor area for one of the principal stimuli of thirst, the octapeptide, angiotensin II. In conscious water-sated rats, the authors examined the effects of intravenous infusion of angiotensin II on the rate of glucose utilization in the subfornical organ and in structures anatomically and functionally connected with it. Angiotensin II produced pressor and drinking responses and increased glucose utilization selectively in the subfornical organ and pituitary neural lobe and in no other brain structure. Treatment with the angiotensin II antagonist, sar1-leu8-angiotensin II, before intravenous administration of angiotensin II prevented metabolic stimulation of the subfornical organ and neural lobe. Captopril, an inhibitor of angiotensin-converting enzyme, reduced subfornical organ glucose metabolism to a level similar to that found in control animals. These results demonstrate that peripheral angiotensin II stimulates glucose ...

1985-01-01

365

Recognition and Detoxification of the Insecticide DDT by Drosophila melanogaster Glutathione S-Transferase D1

Selenium and iodine are trace elements that play an important role in health of man and animals. Concerning supply of both elements it is important to meet nutritional needs. According to discoveries selenium is a part of an active group of glutathione peroxidase enzyme. It prevents necrotic changes in liver, muscle dystrophy and has impact on growth. Extensive studies on content of selenium in plant and animal origin products were started in the beginning of the seventies in Western Europe (Frankenberger, et al., 1998). The aim of our work was to determine selenium and iodine content in total diets of Slovene military. Slovene military nutrition is based on its own nutritional standards, which determine 200 #mu#g of iodine and 50 #mu#g of selenium in daily diet (Rep. Slovenija Ministrstvo za Obrambo, 1994). To determine if the amounts of both elements meet the standards we have analysed 20 total daily diets from four different barracks within a two month period. ...

2003-06-10

366

Radiation processing in Hungary

GSTD1 is one of several insect glutathione S-transferases capable of metabolizing the insecticide DDT. Here we use crystallography and NMR to elucidate the binding of DDT and glutathione to GSTD1. The crystal structure of Drosophila melanogaster GSTD1 has been determined to 1.1 {angstrom} resolution, which reveals that the enzyme adopts the canonical GST fold but with a partially occluded active site caused by the packing of a C-terminal helix against one wall of the binding site for substrates. This helix would need to unwind or be displaced to enable catalysis. When the C-terminal helix is removed from the model of the crystal structure, DDT can be computationally docked into the active site in an orientation favoring catalysis. Two-dimensional {sup 1}H,{sup 15}N heteronuclear single-quantum coherence NMR experiments of GSTD1 indicate that conformational changes occur upon glutathione and DDT binding and the residues that broaden upon DDT binding support the ...

2010-06-14

367

Radiation decontamination of dry food ingredients and processing aids

Hungary has 10.7 million population in 100,000 km/sup 2/ territory. The gross national product is about $3,000 per capita per year. Hungary is a country with highly developed agriculture and medium degree developed industries. The Hungarian economy is an open economy because more than 40% of the national income is earned by export. The research and development works on various radiation processing have been performed for 25 years. In the Central Research Institute for Physics of the Hungarian Academy of Sciences, a laboratory was organized for the basic research of radiation chemistry and the moderator materials for nuclear reactors. Also the activities in the Central Research Institute for Chemistry, the Institute of Isotopes, the Research Institute for Plastics Industry, and the Central Research Institute for Food Industry are briefly reported. The largest radiation processing unit in Hungary is the automatic sterilization plant of Medicor Works in Debrecen with 350 kCi Co-60 source. ...

1982-01-01

368

Proteomic analysis of the shistosome tegument and its surface membranes

Radiation decontamination of dry ingredients, herbs and enzyme preparations is a technically feasible, economically viable and safe physical process. The procedure is direct, simple, requires no additives and is highly efficient. Its dose requirement is moderate. Radiation doses of 3-10 kGy (0.3-1 mrad) have proved sufficient to reduce the viable counts to a satisfactory level. Ionising radiations do not cause any significant rise in temperature. The flavour, texture or other important technological or sensory properties of most ingredients are not influenced at radiation doses necessary for satisfactory decontamination, and radiation obviates the chemical residue problem. The microflora surviving radiation decontamination of dry ingredients are more susceptible to subsequent antimicrobial treatments. Recontamination can be prevented as the product can be irradiated in its final packaging. Irradiation could be carried out in commercial containers and would result ...

369

Scientific Electronic Library Online (English)

Abstract in english The tegument surface of the adult schistosome, bounded by a normal plasma membrane overlain by a secreted membranocalyx, holds the key to understanding how schistosomes evade host immune responses. Recent advances in mass spectrometry (MS), and the sequencing of the Schistosoma mansoni transcriptome/genome, have facilitated schistosome proteomics. We detached the tegument from the worm body and enriched its surface membranes by differential extraction, before subjecting t (more) he preparation to liquid chromatography-based proteomics to identify its constituents. The most exposed proteins on live worms were labelled with impearmeant biotinylation reagents, and we also developed methods to isolate the membranocalyx for analysis. We identified transporters for sugars, amino acids, inorganic ions and water, which confirm the importance of the tegument plasma membrane in nutrient acquisition and solute balance. Enzymes, including phosphohydrolases, ...

2006-10-01

370

Proteins of the kidney microvillar membrane

Production of a cellulolytic enzyme system in mixed-culture solid-state fermentation of soybean hulls supplemented with wheat bran

Two methods were used to label pig kidney microvillar membrane proteins from the luminal and cytoplasmic surfaces of closed membrane vesicles. The first method was lactoperoxidase-catalysed radioiodination. Lactoperoxidase and glucose oxidase were positioned inside or outside the vesicles, iodination being initiated by adding glucose and "1"2"5I. After electrophoresis of the proteins, asymmetric labelling patterns on radioautographs were observed. However the major disadvantage of this method was the high degree of intramembrane labelling of the fatty acid chains of membrane lipids. The second method overcame this disadvantage. A new hydophilic photoreagent, 3,5-di("1"2"5I)iodo-4-azidobenzenesulphonate, was transported by a Na"+-dependent system into microvillar vesicles, thus permitting labelling from either side of the membrane when the vesicles were photolysed. The activity of several microvillar peptidases survived the labelling reaction and they could be identified in the ...

371

British Library Electronic Table of Contents (United Kingdom)

Solid-state fermentation of soybean hulls supplemented with wheat bran using a co-culture of Trichoderma reesei and Aspergillus oryzae was performed. Three parameters - initial moisture content, incubation temperature, and initial pH - were optimized in culture flasks using response surface methodology. Parameter optimization was carried out with respect to filter paper activity and b-glucosidase activity in the culture. Temperature of 30^oC, pH of 5, and moisture content of 70% were found to be optimum. Optimized parameters were used for laboratory scale-up in static tray fermenters. The maximum filter paper activity of 10.7FPU/g-ds and b-glucosidase of 10.7IU/g-ds were obtained after 96-h incubation period in static tray fermenters in agreement with optimized activities at shake flask le...

2010-01-01

372

Preferential induction of cytochrome P450 1A1 over cytochrome P450 1B1 in human breast epithelial cells following exposure to quercetin

British Library Electronic Table of Contents (United Kingdom)

Estrogen metabolism is suggested to play an important role in estrogen-induced breast carcinogenesis. Epidemiologic studies suggest that diets rich in phytoestrogens are associated with a reduced risk of breast cancer. Phytoestrogens are biologically active plant compounds that structurally mimic 17b-estradiol (E2). We hypothesize that phytoestrogens, may provide protection against breast carcinogenesis by altering the expression of estrogen-metabolizing enzymes cytochrome P450 1A1 (Cyp1A1) and 1B1 (Cyp1B1). Cyp1A1 and Cyp1B1 are responsible for the metabolism of E2 to generate 2-hydroxyestradiol (2-OHE2) and 4-hydroxyestradiol (4-OHE2), respectively. Studies suggest that 2-OHE2 and 2-methoxyestradiol may protect against breast carcinogenesis, while 4-OHE2 is carcinogenic in rodent models....

2008-01-01

373

Organisms posses enzymes that function in the repair of DNA damaged by radiations, chemicals and metabolic events

Modelling 18O2 and 16O2 unidirectional fluxes in plants: I. Regulation of pre-industrial atmosphere

This report briefly describes the studies on the mechanism of in vivo DNA repairing by the author in Research Reactor Institute, Kyoto Univ. for the past 30 years. First, the ability of UV radiation to induce transformation was investigated with viral DNA. The formation of thymine-thymine dimer was found harmful to organisms and such dimers were removable by UV-radiation at a low frequency. The mutability was determined in three different E.coli strains with mutator gene, mutT, mutS or mutL. The ability to excise 8-oxoguanin developed in primer DNA was deficient in mutT and miss-pairing left after DNA replication could not be recovered in mutL and mutS strains. Further, DNA repairing mechanism was investigated in other microorganisms; single-strand cleavage caused by exposure to BNCB radiation (boron-neutron-captured beam) could not be repaired in E. coli. Whereas for Deinococcus radiodurans, of which survival rate was not decreased by #gamma#-ray radiation at 5 kGy or less, it was ...

1998-01-01

374

British Library Electronic Table of Contents (United Kingdom)

In closed systems, the O2 compensation point (?O) was previously defined as the upper limit of O2 level, at a given CO2 level, above which plants cannot have positive carbon balance and survive. Studies with 18O2 measure the actual O2 uptake by photorespiration due to the dual function of Rubisco, the enzyme that fixes CO2 and takes O2 as an alternative substrate. One-step modelling of CO2 and O2 uptakes allows calculating a plant specificity factor (Sp) as the sum of the biochemical specificity of Rubisco and a biophysical specificity, function of the resistance to CO2 transfer from the atmosphere to Rubisco. The crossing points (Cx, Ox) are defined as CO2 and O2 concentrations for which O2 and CO2 uptakes are equal. It is observed that: (1) under the preindustrial atmosphere, photorespir...

2011-01-01

375

Mechanism of trypsin inactivation by intact Hymenolepis diminuta (Cestoda)

Lipid A biosynthesis in Rhizobium leguminosarum: Role of a 2-keto-3-deoxyoctulosonate-activated 4{prime} phosphatase

The mechanism of trypsin inactivation by intact Hymenolepis diminuta has been investigated by biochemical and autoradiographic methods. Although worms inactivate trypsin and chymotrypsin in vitro, no inactivation of other endoproteases (subtilisin, pepsin and papain) could be demonstrated. Trypsin inactivation, as demonstrated by macromolecular substrates (azoalbumin, hemoglobin and casein), could not be detected using low molecular weight synthetic substrates such as N-p-benzoyl-DL-arginine-p-nitroanilide (BAPA) or N-p-tosyl-L-arginine methyl ester (TAME). In addition, the kinetic parameters (K/sub m/ and k_3) for H. diminuta-inactivated trypsin, using BAPA as the substrate, were not different from those of the native enzyme. The number of active sites for both native and inactivated trypsin were determined by titration with p-nitro-phenyl-p'-guanidinobenzoate. Absorbance values for both titrations were found to be identical. Dialysis, heating to 50 C, or repeated ...

1979-01-01

376

Intracellular monitoring of superoxide dismutase expression in an Escherichia coli fed-batch cultivation using on-line disruption with at-line surface plasmon resonance detection.

Lipid A from several strains of the N{sub 2}-fixing bacterium Rhizobium leguminosarum displays significant structural differences from Escherichia coli lipid A, one of which is the complete absence of phosphate groups. However, the first seven enzymes of E. coli lipid A biosynthesis, leading from UDP-GlcNAc to the phosphorylated intermediate, 2-keto-3-deoxyoctulosonate (Kdo{sub 2})-lipid IV{sub A}, are present in R. leguminosarum. We now describe a membrane-bound phosphatase in R. leguminosarum extracts that removes the 4{prime} phosphate of Kdo{sub 2}-lipid IV{sub A}. The 4{prime} phosphatase is selective for substrates containing the Kdo domain. It is present in extracts of R. leguminosarum biovars phaseoli, viciae, and trifolii but is not detectable in E. coli and Rhizobium meliloti. A nodulation-defective strain (24AR) of R. leguminosarum bovar trifolii, known to contain a 4{prime} phosphate residue on its lipid A, also lacks measurable 4{prime} phosphatase ...

1995-08-01

377

Inhibition of calmodulin - regulated calcium pump activity in rat brain by toxaphene

An on-line cell disruption system for at-line monitoring of the intracellular concentration of recombinant human superoxide dismutase (rhSOD) in a genetically modified Escherichia coli strain, HMS174(DE3) (pET11a/rhSOD), in bioreactor cultivations is described. The sampled bacteria were disrupted on-line by rapid mixing with a nonionic detergent. The recombinant protein content of the lysed bacterial sample was quantitated by a subsequent surface plasmon resonance biosensor with a specific monoclonal antibody. Extraction efficiency of the monitoring system was optimized with respect to the flow rate ratio of the cell suspension and the detergent at relevant cell densities with the aim to attain rapid monitoring. Monitoring was demonstrated for a shake flask culture and a glucose-limited fed-batch cultivation. The results are compared with a traditional enzyme-linked immunosorbent assay method showing a correlation coefficient of R2 = 0.97. Extraction efficiency of ...

2005-07-01

378

Inhibition of calmodulin - regulated calcium pump activity in rat brain by toxaphene

In vivo effects of toxaphene on calcium pump activity in rat brain synaptosomes was studied. Male Sprague-Dawley rats were dosed with toxaphene at 0,25,50, and 100 mg/kg/day for 3 days and sacrificed 24 h after last dose. Ca/sup 2 +/-ATPase activity and /sup 45/Ca uptake were determined in brain P/sub 2/ fraction. Toxaphene inhibited both Ca/sup 2 +/-ATPase activity and /sup 45/Ca/sup 2 +/ uptake and the inhibition was dose dependent. Both substrate and Ca/sup 2 +/ activation kinetics of Ca/sup 2 +/-ATPase indicated non-competitive type of inhibition as evidenced by decreased catalytic velocity but not enzyme-substrate affinity. The inhibited Ca/sup 2 +/-ATPase activity and Ca/sup 2 +/ uptake were restored to normal level by exogenously added calmodulin which increased both velocity and affinity. The inhibition of Ca/sup 2 +/-ATPase activity and Ca/sup 2 +/ uptake and restoration by calmodulin suggests that toxaphene may impair active calcium transport mechanisms ...

1986-03-05

379

Induction of mutation in Trichoderma viride for conversion of natural cellulose into glucose

In vivo effects of toxaphene on calcium pump activity in rat brain synaptosomes was studied. Male Sprague-Dawley rats were dosed with toxaphene at 0,25,50, and 100 mg/kg/day for 3 days and sacrificed 24 h after last dose. Ca"2"+-ATPase activity and "4"5Ca uptake were determined in brain P_2 fraction. Toxaphene inhibited both Ca"2"+-ATPase activity and "4"5Ca"2"+ uptake and the inhibition was dose dependent. Both substrate and Ca"2"+ activation kinetics of Ca"2"+-ATPase indicated non-competitive type of inhibition as evidenced by decreased catalytic velocity but not enzyme-substrate affinity. The inhibited Ca"2"+-ATPase activity and Ca"2"+ uptake were restored to normal level by exogenously added calmodulin which increased both velocity and affinity. The inhibition of Ca"2"+-ATPase activity and Ca"2"+ uptake and restoration by calmodulin suggests that toxaphene may impair active calcium transport mechanisms by decreasing regulator protein calmodulin levels.

1986-04-13

380

Increased transcription of the c-myc oncogene in two methylcholanthrene-induced quail fibroblastic cell lines

The production of cellulolytic enzymes from fungi has been extensively studied. Several mutants of Trichoderma reesei were selected. Most of the studies were carried out on T. reesei, T. viride, T. harzianum, Penicillium funiculosum, Altemaria alternata. Aspergillus phoenicis, A. ustus, A. tamarii, A. japonicus, and A. niger. T. koningii is one of the most active producers of the so-called C, factor, which is indispensable for the rapid and extensive attack on crystal-line cellulose. However, Trichodenna is known to excrete only small amounts of {beta}-glucosidase. Therefore, Trichoderma is supplemented with {beta}-glucosidase from Aspergillus to increase the saccharification rate of cellulose to glucose as the main sugar. Induction of mutations in Trichodenna spp. rather than T. viride as a tool for the enhancement of {beta}-glucosidase activity was reported. Unfortunately, T. reesei is a poor producer of {beta}-glucosidase. On the other hand, T. harzianum M{sub ...

1991-12-31

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381

In-situ synthesis of poly(dimethylsiloxane)-gold nanoparticles composite films and its application in microfluidic systems.

The expression of three c-onc genes (c-erb, c-myc, c-myb) was investigated in five cell lines established from fibrosarcomas induced with 20-methylcholanthrene (MCA) of Japanese quails. These cell lines showed low levels of the three c-onc genes, with the exception of two cell lines that accumulated moderate (MCAQ 1-4) and large amounts (MCAQ 3-5) of c-myc RNA. Molecular cloning and restriction endonuclease analyses indicated that expression of c-myc in these two cell lines were not associated with detectable rearrangements in the c-myc locus, that the size of the c-myc transcript (2.7 kb) in MCAQ 3-5 was similar to that of the normal c-myc messenger RNAs (mRNA) and that the transcriptional activatin observed in MCAQ 3-5 was not mediated by the LTR (long terminal repeat) of a proximate ALV (avian leukosis virus) provirus. Finally, when analyzed with the restriction enzymes Msp I and Hpa II, the c-myc locus of MCAQ 3-5 and MCAQ 1-4 was found hypomethylated as ...

1984-12-01

382

In vivo effects of cadmium chloride on certain aspects of protein metabolism in tissues of a freshwater field crab Barytelphusa guerini

We presented a simple approach for in-situ synthesis of poly(dimethylsiloxane) (PDMS)-gold nanoparticles composite film based on the special characteristics of PDMS itself. It is an environmentally safe synthesis method without the requirement of additional reducing/stabilizing agents. The region where the resulting gold nanoparticles distribute (in the matrix or on the surface of the polymer) and the size of the nanoparticles, as well as the colour of the free-standing films, can be simply controlled by adjusting the ratio of curing agent and the PDMS monomer. The chemical and optical properties of these composite films were studied. Using such a method, gold nanoparticle micropatterns on PDMS surfaces can be performed. And based on the gold nanoparticles micropattern, further modification with antibodies, antigens, enzymes and other biomolecules can be achieved. To verify this ability, an immobilized glucose oxidase (GOx) reactor in microchannels was built and ...

2007-11-14

383

In vivo effects of cadmium chloride on certain aspects of carbohydrate metabolism in the tissues of a freshwater field crab Barytelphusa guerini

Heavy metals discharged from industries are a major source of pollution which has become a threat to all forms of life. Among the various heavy metals, cadmium is known to be highly toxic even in low concentrations. The harmful effects of cadmium is attributed to its effects on sulfhydryl groups of enzymes, especially dehydrogenases. Various aspects of cadmium pollution on fishes have been extensively reviewed. A survey of the literature reveals that few attempts have been made to study the various aspects of cadmium toxicity in crustaceans and these studies were mainly devoted to marine forms. The freshwater crustaceans, particularly the freshwater field crab, Barytelphusa guerini, has received little attention. The present study reports the influence of cadmium on certain aspects of protein metabolism in the tissues of a freshwater field crab, Barytelphusa guerini, and important component of the paddy field ecosystem, exposed to sublethal concentrations of ...

1991-04-01

384

Human cytoplasmic actin proteins are encoded by a multigene family

Cadmium is a toxic, non-essential heavy metal inhibiting numerous enzymes with functional sulfhydryl groups. Among the animals, aquatic organisms are most sensitive to heavy metals. Various aspects of toxic effects of cadmium pollution on fishes have been extensively reviewed. Survey of literature reveals that relatively few attempts have been made on the various aspects of cadmium toxicity in crustaceans and these studies were mainly devoted to marine forms. The freshwater crustaceans, particularly the freshwater field crab, Barytelphusa guerini, received less attention. This crab forms one of the major components of the paddy field ecosystem and has an edible importance among local populations. Apart from this, these crabs are easily available, maintainable in the laboratory and data obtained in this study can be extrapolated to other crustaceans. The present study reports the influence of cadmium on certain aspects of carbohydrate metabolism in the tissues of ...

1989-06-01

385

Genetic engineering of group 2 sigma factor SigE widely activates expressions of sugar catabolic genes in Synechocystis species PCC 6803.

The authors characterized nine human actin genes that they isolated from a library of cloned human DNA. Measurements of the thermal stability of hybrids formed between each cloned actin gene and ..cap alpha..-, ..beta..-, and ..gamma..-actin mRNA demonstrated that only one of the clones is most homologous to sarcomeric actin mRNA, whereas the remaining eight clones are most homologous to cytoplasmic actin mRNA. By the following criteria they show that these nine clones represent nine different actin gene loci rather than different alleles or different parts of a single gene: (i) the restriction enzyme maps of the coding regions are dissimilar; (ii) each clone contains sufficient coding region to encode all or most of an entire actin gene; and (iii) each clone contains sequences homologous to both the 5' and 3' ends of the coding region of a cloned chicken ..beta..-actin cDNA. They conclude, therefore, that the human cytoplasmic actin proteins are ...

1982-06-01

386

Gene discovery in the Acanthamoeba castellanii genome

Metabolic engineering of photosynthetic organisms is required for utilization of light energy and for reducing carbon emissions.Control of transcriptional regulators is a powerful approach for changing cellular dynamics, because a set of genes is concomitantly regulated. Here, we show that overexpression of a group 2 ? factor, SigE, enhances the expressions of sugar catabolic genes in the unicellular cyanobacterium, Synechocystis sp. PCC 6803. Transcriptome analysis revealed that genes for the oxidative pentose phosphate pathway and glycogen catabolism are induced by overproduction of SigE. Immunoblotting showed that protein levels of sugar catabolic enzymes, such as glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, glycogen phosphorylase, and isoamylase, are increased. Glycogen levels are reduced in the SigE-overexpressing strain grown under light. Metabolome analysis revealed that metabolite levels of the TCA cycle and acetyl-CoA are ...

2011-07-11

387

Evaluation of the radioprotective effect of turmeric extract and vitamin E in mice exposed to therapeutic dose of radioiodine

Acanthamoeba castellanii is a free-living amoeba found in soil, freshwater, and marine environments and an important predator of bacteria. Acanthamoeba castellanii is also an opportunistic pathogen of clinical interest, responsible for several distinct diseases in humans. In order to provide a genomic platform for the study of this ubiquitous and important protist, we generated a sequence survey of approximately 0.5 x coverage of the genome. The data predict that A. castellanii exhibits a greater biosynthetic capacity than the free-living Dictyostelium discoideum and the parasite Entamoeba histolytica, providing an explanation for the ability of A. castellanii to inhabit adversity of environments. Alginate lyase may provide access to bacteria within biofilms by breaking down the biofilm matrix, and polyhydroxybutyrate depolymerase may facilitate utilization of the bacterial storage compound polyhydroxybutyrate as a food source. Enzymes for the synthesis and ...

2005-08-01

388

Erratum to: Inhibition of triclabendazole metabolism in vitro by ketoconazole increases disruption to the tegument of a triclabendazole-resistant isolate of Fasciola hepatica

The aim of this study was to evaluate the radioprotective effect of turmeric extract (40 mg/kg body weight) and vitamin E (#alpha# - tocopherol acetate, 400 IU/kg body weight) supplementation on lipid peroxidation, reduced glutathione and antioxidant defense enzymes in various organs like liver, kidney and salivary glands at 24 h in adult Swiss mice. "1"3"1Iodine exposure significantly increased lipid peroxidation in kidney and salivary glands in comparison to control animals. Pre supplementation with turmeric extract for 15 days showed significant lowering of lipid peroxidation in kidney. On the other hand vitamin E pre supplementation showed marked reduction in lipid peroxidation in salivary glands. Reduced glutathione levels decreased significantly in liver after radiation exposure. However, pre supplementation with turmeric extract and vitamin E did not improve glutathione levels in liver. In conclusion we have observed differential radioprotective effect of ...

2008-10-01

389

British Library Electronic Table of Contents (United Kingdom)

A study has been carried out to investigate whether the action of triclabendazole (TCBZ) against Fasciola hepatica is altered by inhibition of drug metabolism. The cytochrome P450 (CYP 450) enzyme pathway was inhibited using ketoconazole (KTZ) to see whether a TCBZ-resistant isolate could be made more sensitive to TCBZ action. The Oberon TCBZ-resistant and Cullompton TCBZ-susceptible isolates were used for these experiments. The CYP 450 system was inhibited by a 2-h pre-incubation in ketoconazole (40??M), then incubated for a further 22?h in NCTC medium containing either KTZ, KTZ?+?nicotinamide adenine dinucleotide phosphate (NADPH) (1?nM), KTZ?+?NADPH?+?TCBZ (15??g/ml), or KTZ?+?NADPH?+?triclabendazole sulphoxide (TCBZ.SO; 15??g/ml). Changes to fluke ultrastructure following drug treatmen...

2011-01-01

390

Enzyme-linked immunosorbent assays for alpha-synuclein with species and multimeric state specificities.

Enzymatic mechanism in low chloride media: influence of glucose oxidase on the electrochemical behaviour of AISI 316L stainless steel; Mecanisme enzymatique en milieux faiblement chlorures: influence de la glucose oxydase sur le comportement electrochimique de l'acier inoxydable AISI 316L

Abnormal intracellular deposition of aggregated ?-synuclein is the characteristic feature of a number of neurological disorders, including Parkinson's disease (PD). Although ?-synuclein is typically known as a cytosolic protein, a small amount is secreted by exocytosis in both monomeric and aggregated forms. The extracellular forms of ?-synuclein in human body fluids, such as cerebrospinal fluid (CSF) and blood plasma, might be a diagnostic target for PD and related diseases. Here, we characterized a new set of monoclonal antibodies against ?-synuclein, and using different combinations of antibodies, we established ELISA systems to specifically detect human ?-synuclein, mouse and human ?-synuclein together, and multimeric forms of ?-synuclein in biological samples. By employing the Tyramide signal amplification method, the sensitivity of the assay was significantly improved to detect a concentration as low as ?12.5 pg/ml. These assays might be useful tools for quantitative analysis of ...

2011-05-31

391

Engineering of a psychrophilic bacterium for the bioremediation of aromatic compounds

The open circuit potential (OCP) of stainless steels immersed in natural waters generally increases with time. This phenomenon is strongly linked to the formation of a bio-film on the surface. Several studies have proved that the bio-film modifies the reaction of reduction of dissolved oxygen and that it acts mainly on the cathodic processes. One of the hypotheses explaining the action of the bio-film involves certain bacteria which use dissolved oxygen and extracellular enzymes to produce reactive oxygen species. Among this kind of metabolites is hydrogen peroxide H{sub 2}O{sub 2}. This compound interacts with the passive layer of stainless steels and affects their electrochemical behaviour, even when the chloride concentration is low. The aim of this work is to study the influence of an enzymatic reaction (of the oxidoreductase type) on anodic and cathodic processes on AISI 316L stainless steel. Experiments are carried out in two artificial electrolytes ...

2006-07-01

392

Endonuclease IV of Escherichia coli is induced by paraquat

Microbial degradation of aromatic hydrocarbons has been studied with the aim of developing applications for the removal of toxic compounds. Efforts have been directed toward the genetic manipulation of mesophilic bacteria to improve their ability to degrade pollutants, even though many pollution problems occur in sea waters and in effluents of industrial processes which are characterized by low temperatures. From these considerations the idea of engineering a psychrophilic microorganism for the oxidation of aromatic compounds was developed.In a previous paper it was demonstrated that the recombinant Antarctic Pseudoalteromonas haloplanktis TAC125 (PhTAC/tou) expressing a toluene-o-xylene monooxygenase (ToMO) is able to convert several aromatic compounds into corresponding catechols. In our work we improved the metabolic capability of PhTAC/tou cells by combining action of recombinant ToMO enzyme with that of the endogenous P. haloplanktis TAC125 laccase-like ...

2010-01-27

393

Effects of biomass-generated producer gas constituents on cell growth, product distribution and hydrogenase activity of Clostridium carboxidivorans P7{sup T}

The addition of paraquat (methyl viologen) to a growing culture of Escherichia coli K-12 led within 1 hr to a 10- to 20-fold increase in the level of endonuclease IV, a DNase for apurinic/apyrimidinic sites. The induction was blocked by chloramphenicol. Increases of 3-fold or more were also seen with plumbagin, menadione, and phenazine methosulfate. H_2O_2 produced no more than a 2-fold increase in endonuclease IV activity. The following agents had no significant effect: streptonigrin, nitrofurantoin, tert-butyl hydroperoxide, #gamma# rays, 260-nm UV radiation, methyl methanesulfonate, mitomycin C, and ascorbate. Paraquat, plumbagin, menadione, and phenazine methosulfate are known to generate superoxide radical anions via redox cycling in vivo. A mutant lacking superoxide dismutase was unusually sensitive to induction by paraquat. In addition, endonuclease IV could be induced by merely growing the mutant in pure O_2. The levels of endonuclease IV in uninduced or paraquat-treated cells ...

394

Effects of an acute dose of gamma radiation exposure on stem diameter growth, carbon gain, and biomass partitioning in Helianthus annuus

In our previous work, we demonstrated that biomass-generated producer gas can be converted to ethanol and acetic acid using a microbial catalyst Clostridium carboxidivorans P7{sup T}. Results showed that the producer gas (1) induced cell dormancy, (2) inhibited H{sub 2} consumption, and (3) affected the acetic acid/ethanol product distribution. Results of this work showed that tars were the likely cause of cell dormancy and product redistribution and that the addition of a 0.025{mu}m filter in the gas cleanup negated the effects of tars. C. carboxidivorans P7{sup T} can adapt to the tars (i.e. grow) only after prolonged exposure. Nitric oxide, present in the producer gas at 150ppm, is an inhibitor of the hydrogenase enzyme involved in H{sub 2} consumption. We conclude that significant conditioning of the producer gas will be required for the successful coupling of biomass-generated producer gas with fermentation to produce ethanol and acetic acid. (author)

2006-07-15

395

Effect of water activity on the lipase catalyzed esterification of geraniol in ionic liquid [bmim]PF6.

Nineteen-day-old dwarf sunflower plants (Helianthus annuus, variety NK894) received a variable dose (0-40 Gy) from a cobalt-60 gamma source. A very sensitive stem monitoring device, developed at Battelle's Pacific Northwest Laboratories, Richland, Washington was used to measure real-time changes in stem diameter. Exposure of plants caused a significant reduction in stem growth and root biomass. Doses as low as 5 Gy resulted in a significant increase in leaf density, suggesting that nonreversible morphological growth changes could be induced by very low doses of radiation. Carbohydrate analysis of 40-Gy irradiated plants demonstrated significantly more starch content in leaves and significantly less starch content in stems 18 days after exposure than did control plants. In contrast, the carbohydrate content in roots of 40-Gy irradiated plants were not significantly different from unirradiated plants 18 days after exposure. These results indicate that radiation either decreased ...

1988-05-25

396

Crystal structure of Spot 14, a modulator of fatty acid synthesis

Enzymatic reactions in non-aqueous media have been shown to be effective in carrying out chemical transformation where the reactants are insoluble in water or water is a byproduct limiting conversion. Ionic liquids, liquid organic salts with infinitesimal vapor pressure, are potentially useful alternatives to organic solvents. It is known that the thermodynamic water activity is an important variable affecting the activity of enzymes in non-aqueous solvents. This study investigated the influence of water activity on the esterification of geraniol with acetic acid in ionic liquid [bmim]PF6 catalyzed by immobilized Candida antarctica lipase B. The conversion of geraniol in [bmim]PF6 was significant although the reaction rate was slower than in organic solvents. The profile of initial reaction rate-water activity was determined experimentally, and differed from the data reported for other non-aqueous solvents. A maximum in the initial reaction rate was found at aw = ...

2006-02-01

397

Coupling of Realistic Rate Estimates with Genomics for Assessing Contaminant Attenuation and Long-Term Plume Containment - Task 4: Modeling - Final Report

Spot 14 (S14) is a protein that is abundantly expressed in lipogenic tissues and is regulated in a manner similar to other enzymes involved in fatty acid synthesis. Deletion of S14 in mice decreased lipid synthesis in lactating mammary tissue, but the mechanism of S14's action is unknown. Here we present the crystal structure of S14 to 2.65 {angstrom} and biochemical data showing that S14 can form heterodimers with MIG12. MIG12 modulates fatty acid synthesis by inducing the polymerization and activity of acetyl-CoA carboxylase, the first committed enzymatic reaction in the fatty acid synthesis pathway. Coexpression of S14 and MIG12 leads to heterodimers and reduced acetyl-CoA carboxylase polymerization and activity. The structure of S14 suggests a mechanism whereby heterodimer formation with MIG12 attenuates the ability of MIG12 to activate ACC.

2011-09-06

398

Containment of genetically engineered organisms after application to subsurface environments. Technical completion report

Trichloroethene (TCE), a common groundwater contaminant, can be degraded under certain conditions by microorganisms that occur naturally in the subsurface. TCE can be degraded under anaerobic conditions to less chlorinated compounds and ultimately into the non-chlorinated, non-hazardous end product, ethene, via anaerobic reductive dechlorination (ARD). ARD is widely recognized as a TCE degradation mechanism, and occurs in active groundwater remediation and can occur during monitored natural attenuation (MNA). MNA relies on natural processes, such as dispersion and degradation, to reduce contaminant concentrations to acceptable levels without active human intervention other than monitoring. TCE can also be biodegraded under aerobic conditions via cometabolism, in which microbial enzymes produced for other purposes fortuitously also react with TCE. In cometabolism, TCE is oxidized directly to non-hazardous products. Cometabolism as a TCE-degrading process under ...

2005-10-31

399

Construction of a human MluI YAC library

The feasibility of containing genetically engineered bacteria with enhanced dehalogenating properties for in situ bioremediation was investigated. (1) An agarose matrix microbead protocol and a detection system for contained microorganisms or DNA were developed. Multiplex Polymerase Chain Reaction (PCR) allowed tracking of a consortium of encapsulated organisms or several gene targets from a single species. Gene sequences encoding the enzymes responsible for initiating the biodegradation of toluene, octane, and 2,4-D were detected by multiplex PCR and nucleic acid probes from similar to 1-10 biodegradative cells/g soil. Improved DNA extraction methods resulted in PCR reactions detecting similar to 6 cells/g soil. (2) The pcpB gene (for the broad-spectrum detoxicant pentachlorophenol (PCP) hydroxylase) isolated from Flavobacterium sp. strain ATCC 39723 was used in attempts to develop an improved dehalogenating recombinant microorganism for containment experiments.

1993-06-01

400

Cloning and sequencing of cDNA encoding human DNA topoisomerase II and localization of the gene to chromosome region 17q21-22

The authors describe a cloning strategy for the construction of a human genomic library in yeast artificial chromosomes (YACs) based on complete digestion of high-molecular-weight DNA with the infrequently cutting restriction enzyme MluI. Cloning of MluI fragments in the vector pYAC-RC and one subsequent size fractionation by preparative pulsed-field gel electrophoresis yielded a library with average insert sizes of 600 kb. Ninety-seven percent of the colonies were recombinant. An additional size fractionation of MluI fragments prior to ligation had no significant influence on the size of the YACs. The library currently consists of 5000 clones, which is the equivalent of one human genome. Nineteen percent of the YACs were larger than 1.2 Mb. Since smaller MluI fragments are lost during sizing, they also performed cloning without size fractionation. Only 20% of the colonies were recombinant, probably due to unligated vector fragments that were present during the ...

1994-05-01

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

401

Characterization of mal recombination plasmids cloned in Streptococcus pneumoniae

Two overlapping cDNA clones encoding human DNA topoisomerase II were identified by two independent methods. In one, a human cDNA library in phage {lambda} was screened by hybridization with a mixed oligonucleotide probe encoding a stretch of seven amino acids found in yeast and Drosophila DNA topoisomerase II; in the other, a different human cDNA library in a {lambda}gt11 expression vector was screened for the expression of antigenic determinants that are recognized by rabbit antibodies specific to human DNA topoisomerase II. The entire coding sequences of the human DNA topoisomerase II gene were determined from these and several additional clones, identified through the use of the cloned human TOP2 gene sequences as probes. Hybridization between the cloned sequences and mRNA and genomic DNA indicates that the human enzyme is encoded by a single-copy gene. The location of the gene was mapped to chromosome 17q21-22 by in situ hybridization of a cloned fragment to ...

1988-10-01

402

Celastrol regulates innate immunity response via NF-@kB and Hsp70 in human retinal pigment epithelial cells

The malM locus of Streptococcus pneumoniae was cloned into one of the two PstI sites of the multicopy S. pneumoniae plasmid pMV158. To eliminate chromosomal transformants in the simultaneous selection for tetracycline resistance (coded by pMV158) and maltose utilization, the host cells contained a chromosomal deletion of the mal gene cluster. Two clones were isolated; one with a 3.3 kb insert (pLS70) which behaved like wild type with respect to maltose utilization, and another with a 2.9 kb insert (pLS69) which behaved as though it contained a down promoter mutation. Preliminary mapping of these clones by restriction analysis placed the 0.4kb deletion on a HindIII fragment in the interior of the chromosomal insert. The recombinant plasmids were able to transform over 50% of a recipient population to Mal/sup +/. Enzyme measurements of the clones indicated an overproduction of amylomaltase, constituting up to 10% of the total cellular protein, and supported the ...

1981-01-01

403

British Library Electronic Table of Contents (United Kingdom)

Elevated nuclear factor kappa B (NF-@kB) activity and interleukin-6 (IL-6) secretion participates in the pathology of several age and inflammatory-related diseases, including age-related macular degeneration (AMD), in which retinal pigment epithelial cells are the key target. Recent findings reveal that heat shock protein 70 (Hsp70) may affect regulation of NF-@kB. In the current study, effects of Hsp70 expression on NF-@kB RelA/p65 activity were evaluated in human retinal pigment epithelial cells (ARPE-19) by using celastrol, a novel anti-inflammatory compound. Anti-inflammatory properties of celastrol were determined by measuring expression levels of IL-6 and endogenous NF-@kB levels during lipopolysaccharide (LPS) exposure by using enzyme-linked immunosorbent assays (ELISA). Cell viabil...

2011-01-01

404

Biotechnology developments in the livestock sector in developing countries.

Bioscavenger for protection from toxicity of organophosphorus compounds.

Global meat and milk consumption is exponentially increasing due to population growth, urbanization and changes in lifestyle in the developing world. This is an excellent opportunity for developing countries to improve the livestock sector by using technological advances. Biotechnology is one of the avenues for improved production in the "Livestock revolution". Biotechnology developments applied to livestock health, nutrition, breeding and reproduction are improving with a reasonable pace in developing countries. Simple bio-techniques such as artificial insemination have been well implemented in many parts of the developing world. However, advanced technologies including transgenic plant vaccines, marker assisted selection, solid state fermentation for the production of fibrolytic enzymes, transgenic fodders, embryo transfer and animal cloning are confined largely to research organizations. Some developing countries such as Taiwan, China and Brazil have considered ...

2010-01-01

405

An immunochemical approach to the study of DNA damage and repair. Technical progress report, May 1, 1989--April 30, 1992

Current antidotal regimens for organophosphorus compound (OP) poisoning consist of a combination of pretreatment with a spontaneously reactivating AChE inhibitor such as pyridostigmine bromide, and postexposure therapy with anticholinergic drugs such as atropine sulfate and oximes such as 2-PAM chloride (Gray, 1984). Although these antidotal regimens are effective in preventing lethality of animals from OP poisoning, they do not prevent postexposure incapacitation, convulsions, performance deficits, or, in many cases, permanent brain damage (Dunn and Sidell, 1989). These problems stimulated the development of enzyme bioscavengers as a pretreatment to sequester highly toxic OPs before they reach their physiological targets. Several studies over the last two decades have demonstrated that exogenously administered human serum butyrylcholinesterase (Hu BChE) can be used successfully as a safe, efficacious, and single prophylactic treatment to counteract the toxicity of ...

2006-01-01

406

An immunochemical approach to the study of DNA damage and repair

The overall objective of this project has been to develop immunochemical methods to quantitate unique DNA base damages in order to facilitate studies on radiation-induced damage production and repair. Specifically, we have been using antibodies raised to damaged bases to quantitate unique lesions in model systems in order to evaluate their potential biological consequences. Our approach has been to synthesize modified nucleotides or nucleosides, conjugate them to protein carriers, and use the conjugates as immunogens in rabbits or to prepare monoclonal antibodies. We have been studying damages that are stable radiolysis products found in X-irradiated DNA and thus of potential biological consequence. Our aim is to build an in vitro and in vivo data base on the interactions between model DNA lesions and such cellular enzymes as DNA polymerases and repair endonucleases. Initial studies have focused on pyrimidine ring saturation products (thymine glycol.and ...

1992-05-01

407

An enzyme-amplified microtiter plate assay for ethanol: Its application to the detection of peanut ethanol and alcohol dehydrogenase

The overall objective of this project has been to develop immunochemical methods to quantitate unique DNA base damages in order to facilitate studies on radiation-induced damage production and repair. Specifically, we have been using antibodies raised to damaged bases to quantitate unique lesions in model systems in order to evaluate their potential biological consequences. Our approach has been to synthesize modified nucleotides or nucleosides, conjugate them to protein carriers, and use the conjugates as immunogens in rabbits or to prepare monoclonal antibodies. We have been studying damages that are stable radiolysis products found in X-irradiated DNA and thus of potential biological consequence. Our aim is to build an in vitro and in vivo data base on the interactions between model DNA lesions and such cellular enzymes as DNA polymerases and repair endonucleases. Initial studies have focused on pyrimidine ring saturation products (thymine glycol.and ...

1992-05-01

408

Ammonia Fiber Expansion (AFEX) Pretreatment, Enzymatic Hydrolysis, and Fermentation on Empty Palm Fruit Bunch Fiber (EPFBF) for Cellulosic Ethanol Production

A calorimetric microliter plate assay for ethanol amplified by aldehyde dehydrogenase (ALDH) was developed. In the assay ethanol from a sample took part in a chain-reaction catalyzed by alcohol dehydrogenase (ADH) and amplified by ALDH in the presence of NAD{sup +}, diaphorase, and p-ibdonitrotetrazolium-violet (INT-violet)(a precursor of red product). The resultant reaction gave a red color, the intensity of which was proportional to the amount of ethanol present. Using the technique, the content of activity from peanuts of differing maturity and curing stages were determined respectively. Data showed that immature peanuts had a higher level of ethanol and a lower ADH activity than mature peanuts, and that the level of ethanol and ADH activity decreased with the curing time. This indicates that peanut maturity and curing have an effect on ethanol. Also, this implies that other peanut volatiles could be affected in the same way as ethanol, a major volatile in peanuts.

1995-12-01

409

British Library Electronic Table of Contents (United Kingdom)

Empty palm fruit bunch fiber (EPFBF), a readily available cellulosic biomass from palm processing facilities, is investigated as a potential carbohydrate source for cellulosic ethanol production. This feedstock was pretreated using ammonia fiber expansion (AFEX) and enzymatically hydrolyzed. The best tested AFEX conditions were at 135??C, 45?min retention time, water to dry biomass loading of 1:1 (weight ratio), and ammonia to dry biomass loading of 1:1 (weight ratio). The particle size of the pretreated biomass was reduced post-AFEX. The optimized enzyme formulation consists of Accellerase (84??L/g biomass), Multifect Xylanase (31??L/g biomass), and Multifect Pectinase (24??L/g biomass). This mixture achieved close to 90% of the total maximum yield within 72?h of enzymatic hydrolysis. Fer...

2010-01-01

410

Alteration of the enterohepatic recirculation of bile acids in rats after exposure to ionizing radiation

A procedure for batch separation of sup 14 C-hexose from sup 14 C-sucrose

The aim of this work was to study acute alterations of the enterohepatic recirculation (EHR) of bile acids 3 days after an 8-Gy radiation exposure in vivo in the rat by a washout technique. Using this technique in association with HPLC analysis, the EHR of the major individual bile acids was determined in control and irradiated animals. Ex vivo ileal taurocholate absorption was also studied in Ussing chambers. Major hepatic enzyme activities involved in bile acid synthesis were also measured. Measurements of bile acid intestinal content and intestinal absorption efficiency calculation from washout showed reduced intestinal absorption with significant differences from one bile acid to another: absorption of taurocholate and tauromuricholate was decreased, whereas absorption of the more hydrophobic taurochenodeoxycholate was increased, suggesting that intestinal passive diffusion was enhanced, whereas ileal active transport might be reduced. Basal hepatic secretion ...

2004-02-01

411

Up-regulation of sucrose metabolizing enzymes in Oncidium goldiana grown under elevated carbon dioxide

This presentation describes a method for separating {sup 14}C-hexose from {sup 14}C-sucrose in extracts of plant tissue. Portions of ethanol extracts are treated with activated charcoal in microcentrifuge tubes. Aliquots are removed, ethanol evaporated and replaced with reaction mixture that phosphorylates hexose (HEPPS, K{sub 2}HPO{sub 4}, Mg(C{sub 2}H{sub 3}O{sub 2}){sub 2}, ovalbumen, Na{sub 2}ATP, yeast hexokinase). After a time course, the hexokinase reaction is stopped (slowed considerably) to minimize effects of contamination enzyme activities. The stopping agent used is lyxose, a nonphosphorylable analogue of glucose. The strong anionic charge of phosphate introduced through the hexokinase action results in binding (> 95%) of hexose-phosphate to anion-exchange resin. Sucrose remains unbound (> 95%) in solution. This batch ion-exchange is performed in microcentrifuge tubes to allow many samples to be processed simultaneously. Recovery of radiolabel in ...

1991-05-01

412

Ophidian envenomation strategies and the role of purines.

Experiments were conducted in controlled growth chambers to evaluate how increase in CO{sub 2} concentration affected sucrose metabolizing enzymes, especially sucrose phosphate synthase (SPS; EC 2.4.1.14) and sucrose synthase (SS; EC 2.4.1.13), as well as carbon metabolism and partitioning in a tropical epiphytic orchid species (Oncidium goldiana). Response of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) to elevated CO{sub 2} was determined along with dry mass production, photosynthesis rate, chlorophyll content, total nitrogen and total soluble protein content. After 60 days of growth, there was a 80% and 150% increase in dry mass production in plants grown at 750 and 1100 {mu} l{sup -}1 CO{sub 2}, respectively, compared with those grown at ambient CO{sub 2} (about 370 {mu} l{sup -}1). A similar increase in photosynthesis rate was detected throughout the growth period when measured under growth CO{sub 2} conditions. Concomitantly, there ...

2001-07-01

413

On the impact of low power density microwaves in some living tissues

Snake envenomation employs three well integrated strategies: prey immobilization via hypotension, prey immobilization via paralysis, and prey digestion. Purines (adenosine, guanosine and inosine) evidently play a central role in the envenomation strategies of most advanced snakes. Purines constitute the perfect multifunctional toxins, participating simultaneously in all three envenomation strategies. Because they are endogenous regulatory compounds in all vertebrates, it is impossible for any prey organism to develop resistance to them. Purine generation from endogenous precursors in the prey explains the presence of many hitherto unexplained enzyme activities in snake venoms: 5'-nucleotidase, endonucleases (including ribonuclease), phosphodiesterase, ATPase, ADPase, phosphomonoesterase, and NADase. Phospholipases A(2), cytotoxins, myotoxins, and heparinase also participate in purine liberation, in addition to their better known functions. Adenosine contributes to ...

2002-04-01

414

Vitamins B2 and B6 and genetic polymorphisms related to one-carbon metabolism as risk factors for gastric adenocarcinoma in the European prospective investigation into cancer and nutrition

The biomedical and ecological interest for the microwave impact on the Earth biosphere is continuously increased since the industrial, military and communication activities strongly contribute to the electromagnetic stress of living bodies. In the next the authors present some of the main results obtained regarding the microwave exposure of various types of biological material: bacteria, fungi, young plant seedlings, dry seeds, animal tissues. The electromagnetic exposure was carried out in open space in well controlled environmental conditions by using 10.75 GHz/1 m W cm{sup -2} microwaves. Biochemical assays and cytogenetic tests have been carried out to reveal the changes induced post irradiation. The response of some pathogen bacteria, have been emphasized by means of turbidimetric measurements - the stimulatory effect being noticed at the level of the microbial population density (the stimulation of the human body microbial flora seems to be one of the side effects of microwave ...

2006-07-01

415

Uptake, transport, and storage of calcium and magnesium in spruce (Picea abies [L]Karst.) and pine (Pinus silvestris L.) as affected by variable nutrition and pollutant stress

B vitamins and polymorphisms in genes coding for enzymes involved in one-carbon metabolism may affect DNA synthesis and methylation and thereby be implicated in carcinogenesis. Previous data on vitamins B2 and B6 and genetic polymorphisms other than those involving MTHFR as risk factors for gastric cancer (GC) are sparse and inconsistent. In this case-control study nested within the European Prospective Investigation into Cancer and Nutrition cohort, cases (n = 235) and controls (n = 601) were matched for study center, age, sex, and time of blood sampling. B2 and B6 species were measured in plasma, and the sum of riboflavin and flavin mononucleotide was used as the main exposure variable for vitamin B2 status, whereas the sum of pyridoxal 5'-phosphate, pyridoxal, and 4-pyridoxic acid was used to define vitamin B6 status. In addition, we determined eight polymorphisms related to one-carbon metabolism. Relative risks for GC risk were calculated with conditional ...

2010-01-01

416

The transfer of 2,3,7,8-tetrachlorodibenzo-p-dioxin into eggs and chicks following exposure to hens

Statements about the dynamic processes of uptake, transport, and deposition of Ca and Mg in norway spruce and Scots pine are made in this paper. Concerning the storage of these elements it is shown that there are great differences in their functional importance in cell metabolism. There is evidence that the role of Mg in enzyme and protein metabolism is of far greater significance for the understanding of Mg-deficiency symptoms than its function as the central atom of the chlorophyll complexes. In regard to the transport and especially to the incorporation of Ca into the needles differences between species were evident, expressing the special status of pine among the gymnosperms. With increasing needle age an accumulation of Ca-oxalate crystals, which are physiologically inert, could be proved for the studied conifers. This was interpreted as a 'detoxication' from surplus Ca to hold constant the level of the physiologically active fraction. Accordingly, the low ...

417

The role of the mesenchyme in mouse neural fold elevation. II. Patterns of hyaluronate synthesis and distribution in embryos developing in vitro

Dioxins have been shown to exert reproductive and teratogenic effects in several strains of mice, rats, and chickens. We reported that in ovo exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) at less than 7.5 ng/egg on day 0 did not influence hatchability, whereas more than 10 ng/egg completely inhibited hatching. We also reported that maternal exposure to TCDD in Barred Plymouth Rock hens induced a reversible inhibition of egg laying. The hatchability of the eggs from TCDD exposed hens was significantly decreased and eggshell thickness was thicker than that from control hens 1. These results suggested that the TCDD in maternally exposed hens was transferred into eggs and induced embryo toxicity. Transfer of TCDD in eggs has been reported previously in foraging chickens 2,3 and ring-necked pheasants 4,5. The TCDD concentration in chicken eggs related to environmental exposure, especially contact with soil. The measurement of dioxins in eggs is important for assessing environmental ...

2004-09-15

418

The Sorcerer II Global Ocean Sampling Expedition: Expanding theUniverse of Protein Families

Hyaluronate (HA) distribution patterns were examined in the cranial mesenchyme underlying the mesencephalic neural folds of mouse embryos maintained in roller tube culture. Using standard image-processing techniques, the digitized images of Alcian blue-stained or 3H-glucosamine-labeled sections digested with an enzyme specific for HA, were subtracted from adjacent, undigested sections. The resultant difference picture images (DPI) accurately depicted the distribution of stained or labeled HA within the cranial mesenchyme. 3H-glucosamine-labeled HA was distributed uniformly throughout the cranial mesenchyme as 12, 18, and 24 hr of culture. By contrast, the mesenchyme was uniformly stained with Alcian blue at 12 hr, but stain intensity decreased in the central regions of the mesenchyme at 18 and 24 hr. HA distribution patterns were also examined in the cranial mesenchyme of embryos cultured in the presence of diazo-oxo-norleucine (DON), a glutamine analogue that ...

1990-06-01

419

The Protective Effect of Antioxidants on Oxidative Stress in Rats Exposed to the 950 MHz Electromagnetic Field

Metagenomics projects based on shotgun sequencing of populations of micro-organisms yield insight into protein families. We used sequence similarity clustering to explore proteins with a comprehensive dataset consisting of sequences from available databases together with 6.12 million proteins predicted from an assembly of 7.7 million Global Ocean Sampling (GOS) sequences. The GOS dataset covers nearly all known prokaryotic protein families. A total of 3,995 medium- and large-sized clusters consisting of only GOS sequences are identified, out of which 1,700 have no detectable homology to known families. The GOS-only clusters contain a higher than expected proportion of sequences of viral origin, thus reflecting a poor sampling of viral diversity until now. Protein domain distributions in the GOS dataset and current protein databases show distinct biases. Several protein domains that were previously categorized as kingdom specific are shown to have GOS examples in other kingdoms. About ...

2006-03-23

420

Tests for the diagnosis of Clostridium difficile infection: the next generation.

Studies have linked cell phone radiation to health problems such as headaches, high blood pressure, cancer and more. There is a latency period for most diseases and it may take years and more studies before the required weight of evidence is established. But the effects are cumulative and precautions should be taken now before it is too late. The aim of the present study was to investigate if supplementation with antioxidants would protect heart and liver tissues from harmful radiation emitted by cell phone. Thirty two male albino rats were randomly divided into four equal groups: I- Control, II- Antioxidants treated group, III- 950 MHz EMR, IV- 950 MHz EMR + antioxidants. A 950 MHz EMR radiation (217-Hz pulse rate, 2-W maximum peak power, SAR Specific Absorption Rate"1.6 W/Kg) was applied to groups III and IV 60 min/day, for 30 days using an experimental exposure device. Antioxidants supplement (Vitamins A, E and C + Se) was administered to rats daily, by gavages, during the period of ...

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

421

Synthesis, characterization and biologic effects of polybrominated naphthalenes

Clostridium difficile (C. difficile) causes 25-30% of cases of antibiotic associated diarrhea and most cases of pseudomembranous colitis. Patients presenting with diarrhea after hospitalization for 3 or more days should be tested for C. difficile. There are many options available for testing, each of which has inherent advantages and disadvantages. Most laboratories perform toxin testing using an enzyme immunoassay method. In general these tests have sensitivities ranging from 60 to 70% and specificities of 98%. When using these methods, symptomatic patients with negative tests should be tested by another more sensitive method. Until recently, cell culture cytotoxicity neutralization assays (CCNAs) were considered the gold standard in the U.S. A two-step algorithm using an EIA for glutamate dehydrogenase detection followed by testing positives using CCNA, offered an improved alternative until the availability of molecular assays. Although early studies that ...

2011-03-03

422

Survey report for fiscal 1998. Survey of the current state and tasks of research and development of technologies for effectively utilizing CO{sub 2} fixation by higher vegetation; 1998 nendo chosa hokokusho. Koto shokubutsu ni okeru nisanka tanso koteika yuko riyo gijutsu no kenkyu kaihatsu no genjo to kadai ni kansuru chosa

Although polybrominated naphthalenes (PBNs) are contaminants of the commercial fire retardant fireMaster BP-6, the individual PBN isomers have not been identified. In order to study PBNs possessing an analogous level of bromination to those found in fireMaster BP-6, three synthetic PBN mixtures, averaging 5.0, 5.3, and 5.6 bromines per naphthalene were synthetized and partially characterized. The PBN mixtures were administered to immature male Wistar rats and found to be potent inducers of cytochrome P-450-dependent monooxygenases. At the lowest dose tested, 30 mumol X kg-1, each PBN mixture caused maximal induction of benzo(a)pyrene hydroxylase activity. On the basis of enzyme activities, ligand-binding spectra and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the PBN mixtures were determined to be 3-methylcholanthrene-type inducers of cytochrome P-450 (P-448), resembling qualitatively the most toxic polyhalogenated biphenyls, dibenzofurans, and ...

1984-07-01

423

Structural definition of the active site and catalytic mechanism of 3,4-dihydroxy-2-butanone 4-phosphate synthase

Investigations and studies are conducted seeking for a CO2 fixation method improved by utilizing the photosynthesizing function of higher vegetation. Details of higher vegetation genes are being disclosed thanks to the rapid progress of studies making use of molecular biological techniques, and the application of the genetic mechanism to scientific and technological fields is becoming increasingly feasible. In particular, the role of the CO2 fixation enzyme RuBisCO has been elucidated almost completely. It has been learned that, in terms of photosynthesizing capability, the C{sub 4} plants (corn etc.) are 2-3 times higher than the C{sub 3} plants (rice, wheat, etc.), and 5-10 times higher than the CAM plants (cactuses etc.). Studies are also under way about the rice genome so that a photosynthesizing capability so high as that of the C{sub 4} plants may be endowed the rice plant. The metabolism and control of useful substances produced in the CO2 fixation process ...

1999-03-01

424

Sex and species differences in tyrosine hydroxylase-synthesizing cells of the rodent olfactory extended amygdala.

X-ray crystal structures of L-3,4-dihydroxy-2-butanone-4-phosphate synthase from Magnaporthe grisea are reported for the E-SO{sub 4}{sup 2-}, E-{sub 4}{sup 2-}-Mg{sup 2+}, E-SO{sub 4}{sup 2-}-Mn{sup 2+}, E-SO{sub 4}{sup 2-}-Mn{sup 2+}-glycerol, and E-SO{sub 4}{sup 2-}-Zn{sup 2+} complexes with resolutions that extend to 1.55, 0.98, 1.60, 1.16, and 1.00 {angstrom}, respectively. Active-site residues of the homodimer are fully defined. The structures were used to model the substrate ribulose 5-phosphate in the active site with the phosphate group anchored at the sulfate site and the placement of the ribulose group guided by the glycerol site. The model includes two Mg{sup 2+} cations that bind to the oxygen substituents of the C2, C3, C4, and phosphate groups of the substrate, the side chains of Glu37 and His153, and water molecules. The position of the metal cofactors and the substrate's phosphate group are further stabilized by an extensive hydrogen-bond and salt-bridge ...

2010-03-08

425

Seasonal variation of defense-related gene expression in leaves from Bois noir affected and recovered grapevines.

The bed nucleus of the stria terminalis (BST) and the medial amygdala (MeA) are anatomically connected sites necessary for chemosensory regulation of social behaviors in rodents. Prairie voles (Microtus ochrogaster) are a valuable model for studying the neural regulation of social behaviors because, unlike many other rodents, they are gregarious, pair bond after copulating, and are biparental. We herein describe sex and species differences in immunoreactivity for tyrosine hydroxylase (TH), the rate-limiting enzyme for catecholamine synthesis, in the BST and MeA. Virgin male prairie voles had a large number of TH-immunoreactive cells in areas analogous to the rat principal nucleus of the BST (pBST) and the posterodorsal medial amygdala (MeAPd). Virgin female prairie voles had far fewer TH-immunoreactive cells in these sites ( approximately 17% of the number of cells as males in the pBST, approximately 35% of the number of cells in the MeAPd). A few TH-immunoreactive ...

2007-01-01

426

Role of mitochondria in sulfolipid biosynthesis by Euglena chloroplasts

Although Bois noir is one of the main phytoplasma diseases of grapevine, the gene expression and enzyme activities that underlie physiological changes occurring in symptomatic and recovered (with spontaneous or induced symptom remission) plants are mostly unknown. Bois noir symptomatic leaves (September 2006, 2007) and symptomless leaves from infected symptomatic plants (September 2007) of Sangiovese (moderately susceptible) and Chardonnay (highly susceptible) cultivars were collected. Moreover, leaves from infected symptomless plants of both cultivars were harvested in June 2007. Leaves from recovered plants were also collected in the same periods. In recovered plants of both cultivars, class III chitinase and almost every time phenylalanine ammonia-lyase and chalcone synthase expression were increased for all collection periods. In symptomatic leaves of both cultivars, the expressions of the same genes were up-regulated and also those of ?-1,3-glucanase and ...

2011-05-31

427

Relationship of DNA repair processes to mutagenesis and carcinogenesis in mammalian cells. Progress report, November 1, 1979-October 31, 1980

Sulfate activation occurs in Euglena mitochondria the authors now find that the sulfate activating enzymes are absent from Euglena chloroplasts. Cells of mutant W/sub 10/BSmL lacking plastids also lack detectable sulfolipid (SL) when grown on /sup 35/SO/sub 4//sup 2 -/ indicating that SL is absent from the mitochondria and is exclusively in the plastids. Plastids alone will convert /sup 35/S-cysteine to /sup 35/SL in the presence of ATP and Mg/sup 2 +/; light is stimulatory. Under similar conditions, chloroplasts and mitochondria incubated together convert /sup 35/SO/sub 4//sup 2 -/ to plastid-localized /sup 35/SL but either organelle incubated alone fails to effect this conversion. Unlabeled cysteine blocks SL labeling from sulfate in the mixed incubation; since cysteine is formed from sulfate by Euglena mitochrondria, cysteine (and other compounds) may move from the mitochondrion to the chloroplast to provide the sulfo group for SL formation. Although ...

1987-04-01

428

Regulation of the phosphoinositide pathway in cultured Sertoli cells from immature rats: effects of follicle-stimulating hormone and fluoride

The objective of this research is to determine the role of DNA repair in mutagenesis and carcinogenesis in mammalian cells. Use of the host-cell reactivation viral suicide enrichment procedure was initiated in the isolation of repair-deficient mutants. Lightly mutagenized BHK cells were infected with irradiated Herpes simplex virus (HSV); several radiation-sensitive strains were isolated among the survivors of the infection. The characterization of these strains is progressing and the enrichments are continuing. That alterations in the frequency of mutation of C3H/10T 1/2 cells, occurring as a result of holding the cells in a confluent state following treatment with ethylmethane sulfonate, parallel the alterations in the frequency of neoplastic transformation was found. The repair capabilities of BHK cells were found to be intermediate in comparison to repair-proficient and -deficient human cells with regard to the reactivation of HSV treated with various inactivating agents. The ...

429

Protein oxidation markers in women with and without gestational diabetes mellitus: A possible relation with paraoxonase activity.

Many hormones elicit effects on target cells by stimulating the enzyme phospholipase-C, which catalyzes the hydrolysis of phosphoinositides to the intracellular second messengers diacylglycerol and inositol phosphates. The present study examined the roles of FSH and guanine nucleotide-binding proteins (G-proteins) in regulating the hydrolysis of phosphoinositides in Sertoli cells. Sertoli cell cultures prepared from 16- to 18-day-old rats were incubated for 24 h with myo-[2-3H] inositol to label endogenous phospholipids. Treatment of cells from 0.5-20 min with preparations of ovine FSH ranging in potency from 1-60 times that of NIH FSH S1 did not affect accumulation of inositol phosphates. Levels of total [3H]inositol phosphates [[3H]inositol mono-, di-, and triphosphates (IP, IP2, and IP3)] in FSH-treated cultures was 75-120% the levels in control cultures over the various time intervals studied. Addition of testosterone and the combination of testosterone plus ...

430

Prostaglandin (PG) E3 synthesis elicted by adrenergic stimuli in guinea-pig trachea (GPT) is mediated primarily by B2 adrenergic receptors

AIMS: To clarify the levels of protein oxidation markers such as protein carbonyl (PCO), protein hydroperoxides (P-OOH), advanced oxidation protein products (AOPP) and nitrotyrosine (NT), as well as antioxidative enzymes such as paraoxonase (PON-1) in women with and without gestational diabetes mellitus (GDM). METHODS: The study was conducted on 23 women with GDM and 22 women without GDM. The levels of the P-OOH, AOPP, and PON-1 were determined by colorimetric methods; whereas NT and PCO levels were measured by ELISA. RESULTS: The concentrations of protein oxidation markers were significantly increased and PON1 activity was significantly decreased in GDM group compared to those of normal pregnant women. The control group showed a significant negative correlation between PON-1 and PCO (r=-0.451, p=0.027); whereas in GDM group, there was a significant positive correlation between P-OOH and HbA1c (r=0.89, p=0.001). There was no significant correlation between AOPP, ...

2011-08-30

431

Prostaglandin (PG) E3 synthesis elicted by adrenergic stimuli in guinea-pig trachea (GPT) is mediated primarily by B2 adrenergic receptors

The purpose of this study was to examine arachidonic acid (AA) metabolism and to characterize the type of adrenergic receptor (AR) involved in the production of the major metabolite of this fatty acid. ({sup 14}C)AA was incubated with GPT-rings and the radiolabelled products were extracted and separated by TLC method. The medium was also assayed for radiolabelled immunoreactive PG's (iPG's) and leukotrienes (LT) B4 and C4 by RIA or Enzyme immunoassay (EIA) after exposure to various AR agonists. ({sup 14}C)AA was incorporated into GPT-rings and metabolized mainly into iPGE2 and smaller amounts into PGF2{alpha}. Trace amounts of PGD2 and 6-keto-PGF1{alpha} but not LTB4 or LTC4 were detected by RIA and/or EIA. Incubation of GPT rings for 15 minutes with isoproterenol and salbutamol resulted in a significant increase of PGE2 synthesis (optimum conc: 10{sup {minus}7}, 10{sup {minus}7}M respectively). In contrast, dobutamine, norepinephrine, ...

1990-02-26

432

Part III. Direct enzymatic esterification of lactic acid with fatty acids.

The purpose of this study was to examine arachidonic acid (AA) metabolism and to characterize the type of adrenergic receptor (AR) involved in the production of the major metabolite of this fatty acid. ["1"4C]AA was incubated with GPT-rings and the radiolabelled products were extracted and separated by TLC method. The medium was also assayed for radiolabelled immunoreactive PG's (iPG's) and leukotrienes (LT) B4 and C4 by RIA or Enzyme immunoassay (EIA) after exposure to various AR agonists. ["1"4C]AA was incorporated into GPT-rings and metabolized mainly into iPGE2 and smaller amounts into PGF2#alpha#. Trace amounts of PGD2 and 6-keto-PGF1#alpha# but not LTB4 or LTC4 were detected by RIA and/or EIA. Incubation of GPT rings for 15 minutes with isoproterenol and salbutamol resulted in a significant increase of PGE2 synthesis (optimum conc: 10"-"7, 10"-"7M respectively). In contrast, dobutamine, norepinephrine, phenylnephrine and xylazine (up to 10"-"6M) did not ...

1991-04-21

433

Oriented immobilization of proteins on grafted porous polymers

Lipase catalyzed esterification reactions between lactic acid and several fatty acids have been studied. Difficulties arise in esterifying lactic acid because of the potential for this substance to act both as an acyl donor and as a nucleophile. These difficulties were minimized via strategies which greatly increased the yield of the desired ester. Use of the companion fatty acid in excess with respect to lactic acid in an apolar solvent (n-hexane) in which the lactic is not completely dissolved has been employed to minimize the potential for lactic acid to act as an acyl donor in a self-polymerization reaction.Beneficial and sinergistic effects of both silica gel and molecular sieves on conversion to the desired product are described. However, careful control of the amount of molecular sieves used is required. This fact is a consequence of two opposing effects of this material: i.e. adsorption of both lactic acid and water from the reaction mixture. For reaction between caprylic and ...

2001-07-01

434

Open top chamber experiment ``Edelmannshof``: carbohydrate and energy metabolism in needles and trunks from Norway spruce fumigated with unfiltered or charcoal-filtered air in comparison with untreated trees

The modification of polymers by radiation grafting has been utilized for several decades. The penetrability of gamma rays allows to modify the internal surfaces of porous materials retaining its mechanical properties. In recent years applications of these materials to obtain chromatographic supports and biocatalysts have been reported. In this work, we described the grafting of glycidyl methacrylate (GMA) onto a macroporous polysulfone polymer. Reproducible amount of grafting, from 10% to 60% was obtained by choosing favourable monomer concentration and gamma radiation doses from 6 kGy up. Afterwards, iminodiacetic acid (IDA) and amino phenyl arsine oxide (PAO) were covalently attached to the grafted polyGMA, in correspondence with the grafting degree. Later on, a recombinant histidin-patch thioredoxin protein (HP-rTrx) was immobilized onto this surface by two different ways, involving specific protein orientations. The first one involves an IDA-Ni{sup 2+} complex and three ...

2003-08-01

435

One-step RNA pathogen detection with reverse transcriptase activity of a mutated thermostable Thermus aquaticus DNA polymerase.

As part of a five-year-lasting open-top-chamber experiment at the `Edelmannshof` site (Welzheimer Wald) we analyzed Norway spruce trees (Picea abies (L.) Karst.) with regard to carbohydrate and energy (ATP/ADP ratio) metabolism. End products, intermediates, and a regulator of carbon allocation (starch, sucrose, hexoses, fructose-2,6-bisphosphate) as well as enzyme activities involved in sucrose metabolism (sucrose-P synthase, sucrose synthase, acid invertase) were determined in needle and trunk samples of the different treatments (charcoal-filtered, unfiltered, ambient air). Changes of these parameters were documented annually and with regard to needle development (up to 2 years after bud break), and diurnally during an ozone period in midsummer. Samples from trees grown in an unfiltered atmosphere showed relatively high sucrose and hexose contents of the needles. The capacities of sucrose-P synthase (increased; sucrose synthesis) and acid invertase (decreased; ...

1997-12-01

436

New discoveries in prostate cancer pathogenesis

We describe the cloning and characterization of a mutated thermostable DNA polymerase from Thermus aquaticus (Taq) that exhibits an increased reverse transcriptase activity and is therefore designated for one-step PCR pathogen detection using established real-time detection methods. We demonstrate that this Taq polymerase mutant (Taq M1) has similar PCR sensitivity and nuclease activity as the respective Taq wild-type DNA polymerase. In addition, and in marked contrast to the wild-type, Taq M1 exhibits a significantly increased reverse transcriptase activity especially at high temperatures (>60 degrees C). RNA generally hosts highly stable secondary structure motifs, such as hairpins and G-quadruplexes, which complicate, or in the worst case obviate, reverse transcription (RT). Thus, RT at high temperatures is desired to weaken or melt secondary structure motifs. To demonstrate the ability of Taq M1 for RNA detection of pathogens, we performed TaqMan probe-based diagnostics of ...

2010-02-01

437

Nerve growth factor actions on the brain

Background. Through PSA screening the rate of prostate cancers detected at an early stage has increased significantly; thus a decrease in mortality can be expected in the near future. Despite all scientific efforts, however, the molecular mechanisms underlying the development and progression of prostate cancer remain poorly understood. Prostate cancer is a disease of aging men and epidemiological evidence supports a major contribution to its development through diet, lifestyle and environmental factors. Genetic instability is the basic phenomenon of tissue cell cancerisation. This instability can be hereditary or due to mutations and other chromosomal aberrations acquired during life. In recent years a large number of interesting data have been collected which show the relationships between focal atrophy and genetic instability of the prostate epithelia. Atrophy can be the result of prostatitis, ischemia as well as of oxidative stress (diet). Several chromosomal aberrations typical for ...

438

Molecular evolution and characterization of fungal indoleamine 2,3-dioxygenases.

We examined the effect of the trophic protein, nerve growth factor (NGF), on cultures of fetal rat neostriatum and basal forebrain-medial septal area (BF-MS) to define its role in brain development. Treatment of cultures with NGF resulted in an increase in the specific activity of the cholinergic enzyme choline acetyltransferase (CAT) in both brain areas. CAT was immunocytochemically localized to neurons. In the BF-MS, NGF treatment elicited a marked increase in staining intensity and an apparent increase in the number of CAT-positive neurons. Moreover, treatment of BF-MS cultures with NGF increased the activity of acetylcholinesterase, suggesting that the cholinergic neuron as a whole was affected. To begin defining mechanisms of action of NGF in the BF-MS, we detected NGF receptors by two independent methods. Receptors were localized to two different cellular populations: neuron-like cells, and non-neuron-like cells. Dissociation studies with ["1"2"5I]NGF ...

439

Modification of adenylate cyclase by photoaffinity analogs of forskolin

Indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO) are tryptophan-degrading enzymes. Mammalian IDO expression is induced by cytokines and has antimicrobial and immunomodulatory effects. A major role of mammalian TDO is to supply nicotinamide adenine dinucleotide (NAD(+)). In fungi, the IDO homologue is thought to be expressed constitutively and supply NAD(+), as TDO is absent from their genomes. Here, we reveal the distribution of IDO genes among fungal species and characterize their enzymatic activity. The yeast, Saccharomyces cerevisiae has only one IDO gene, whereas the koji-mold, Aspergillus oryzae has two genes, IDO? and IDO?. The A. oryzae IDO? showed more similar enzymatic properties to those of S. cerevisiae IDO than IDO?, suggesting that the A. oryzae IDO? is a functional homologue of the S. cerevisiae IDO. From the IDO? gene, two isoforms, IDO? and IDO?(+) could be generated by alternative splicing. The latter contained a 17 amino ...

2010-12-18

440

Mechanistic studies of ethylene biosynthesis in higher plants

Photoaffinity labeling analogs of the adenylate cyclase activator forskolin (PF) have been synthesized, purified and tested for their effect on preparations of membrane-bound, Lubrol solubilized and forskolin affinity-purified adenylate cyclase (AC). All analogs of forskolin significantly activated AC. However, in the presence of 0.1 to 0.3 microM forskolin, the less active forskolin photoaffinity probes at 100 microM caused inhibition. This inhibition was dose-dependent for PF, suggesting that PF may complete with F for the same binding site(s). After cross-linking (125I)PF-M to either membrane or Lubrol-solubilized AC preparations by photolysis, a radiolabeled 100-110 kDa protein band was observed after autoradiography following SDS-PAGE. F at 100 microM blocked the photoradiolabeling of this protein. Radioiodination of forskolin-affinity purified AC showed several protein bands on autoradiogram, however, only one band (Mr = 100-110 kDa) was specifically labeled by (125I)PF-M ...

1989-01-01

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

441

In vitro and in vivo activities of T4 endonuclease V mutants altered in the C-terminal aromatic region

Ethylene is a plant hormone that elicits a wide variety of responses in plant tissue. Among these responses are the hastening of abscission, ripening and senescence. In 1979 it was discovered that 1-amino-1-cyclopropane carboxylic acid is the immediate biosynthetic precursor to ethylene. Given the obvious economic significance of ethylene production the authors concentrated their studies on the conversion of ACC to ethylene. They delved into mechanistic aspects of ACC oxidation and they studied potential inhibitors of ethylene forming enzyme (EFE). They synthesized various analogs of ACC and found that EFE shows good stereodiscrimination among alkyl substituted ACC analogs with the 1R, 2S stereoisomer being processed nine times faster than the 1S, 2R isomer in the MeACC series. They also synthesized 2-cyclopropyl ACC which is a good competitive inhibitor of EFE. This compound also causes time dependent loss of EFE activity leading us to believe it is an ...

1986-01-01

442

High-mobility group box 1 release and redox regulation accompany regeneration and remodeling of skeletal muscle.

Genes encoding mutants of the thymine photodimer repair enzyme from bacteriophage T4 (T4 endonuclease V) having an amino acid substitution (T127M, W128A, W128S, Y129A, K130L, Y131A, Y132A) were constructed by use of a previously obtained synthetic gene and expressed in Escherichia coli under the control of the E. coli tryptophan promoter. An in vitro assay of partially fractionated mutant proteins for glycosylase activity was performed with chemically synthesized substrates containing a thymine photodimer. T127M and K130L showed almost the same activity as the wild-type protein. Although W128S, Y131A, and Y132A were slightly active, W128A and Y129A lost activity. The results indicated that the aromatic amino acids around position 130 may be important for the glycosylase activity. Mutant T127M was purified, and the Km value was found to be of the same order as that of the wild type (10(-8) M). In vivo activities for all mutants were characterized with UV-sensitive ...

443

Genome analysis of F. nucleatum sub spp vincentii and its comparison with the genome of F. nucleatum ATCC 25586.

Abstract High-mobility group box 1 (HMGB1), a damage-associated molecular pattern (DAMP) molecules, favors tissue regeneration via recruitment and activation of leukocytes and stem cells. Here we demonstrate, in a model of acute sterile muscle injury, that regeneration is accompanied by active reactive oxygen species (ROS) production counterbalanced and overcome by the generation of antioxidant moieties. Mitochondria are initially responsible for ROS formation. However, they undergo rapid disruption with almost complete disappearance. Twenty-four hours after injury, we observed a strong induction of MURF1 and atrogin-1 ubiquitin ligases, key signals in activation of the proteasome system and induction of muscle atrophy. At later time points, ROS generation is maintained by nonmitochondrial sources. The antioxidant response occurs in both regenerating fibers and leukocytes that express high levels of free thiols and antioxidant enzymes, such as superoxide dismutase ...

2011-05-09

444

Genetic control of immune response to pseudorabies and atrophic rhinitis vaccines: II. Comparison of additive direct and maternal genetic effects.

We present the draft genome sequence and its analysis for Fusobacterium nucleatum sub spp. vincentii (FNV), and compare that genome with F. nucleatum ATCC 25586 (FN). A total of 441 FNV open reading frames (ORFs) with no orthologs in FN have been identified. Of these, 118 ORFs have no known function and are unique to FNV, whereas 323 ORFs have functional orthologs in other organisms. In addition to the excretion of butyrate, H2S and ammonia-like FN, FNV has the additional capability to excrete lactate and aminobutyrate. Unlike FN, FNV is likely to incorporate galactopyranose, galacturonate, and sialic acid into its O-antigen. It appears to transport ferrous iron by an anaerobic ferrous transporter. Genes for eukaryotic type serine/threonine kinase and phosphatase, transpeptidase E-transglycosylase Pbp1A are found in FNV but not in FN. Unique ABC transporters, cryptic phages, and three types of restriction-modification systems have been identified in FNV. ORFs for ethanolamine ...

2003-06-01

445

Formation of silk fibroin nanoparticles in water-miscible organic solvent and their characterization

Data from 52 litters farrowed in two seasons of a cross-fostering experiment were analyzed to estimate variances and covariances for additive direct and maternal genetic effects on immune response to pseudorabies virus and B. bronchiseptica vaccine. Twenty purebred boars and 44 sows of the Duroc, Landrace and Yorkshire breeds were used. Immune response was measured after vaccine challenge. A modified-live pseudorabies (PR) vaccine was administered to piglets at 28 d of age; response was measured by log2 serum neutralization titers at 56 d. An inactivated B. bronchiseptica bacterin was administered at 28, 42 and 112 d. Antibody levels were measured relative to positive and negative controls at 28, 56 and 119 d by using an enzyme-linked immunosorbent assay (ELISA). Results from this study for heritability were .18 +/- .09 for PR titer and .15 +/- .07 and .52 +/- .15 for 56- and 119-d ELISA values, respectively. The variability due to nurse environment (maternal ...

1987-02-01

446

Fiscal 1993 report on results of R and D on innovative technology for producing advanced biomaterial. Peptide applied carbon dioxide fixation/effective utilization technology (First volume); 1993 nendo senshin bio zairyo no sosei kako gijutsu no kenkyu kaihatsu seika hokokusho. 1. Peptide oyo nisanka tanso koteika yuko riyo gijutsu

When Silk fibre derived from Bombyx mori, a native biopolymer, was dissolved in highly concentrated neutral salts such as CaCl{sub 2}, the regenerated liquid silk, a gradually degraded peptide mixture of silk fibroin, could be obtained. The silk fibroin nanoparticles were prepared rapidly from the liquid silk by using water-miscible protonic and polar aprotonic organic solvents. The nanoparticles are insoluble but well dispersed and stable in aqueous solution and are globular particles with a range of 35-125 nm in diameter by means of TEM, SEM, AFM and laser sizer. Over one half of the {epsilon}-amino groups exist around the protein nanoparticles by using a trinitrobenzenesulfonic acid (TNBS) method. Raman spectra shows the tyrosine residues on the surface of the globules are more exposed than those on native silk fibers. The crystalline polymorph and conformation transition of the silk nanoparticles from random-coil and {alpha}-helix form (Silk I) into anti-parallel {beta}-sheet form ...

2007-10-15

447

Extracellular ATP4- promotes cation fluxes in the J774 mouse macrophage cell line

Technology is being developed for preparing functional materials by synthesizing new functional peptides in which non-natural amino acid needed for the functional manifestation is introduced, and by modifying the surface of a base plate such as silica glass by using such peptides. Activities were conducted in the three areas of (1) creation of functional molecules, (2) materialization technology, and (3) comprehensive investigation and research; the activities were carried out independently and parallelly in the first two areas. In (1), design technique for the structures and functions of peptides was developed, as were conformational control technique, synthesis of peptides having optical/electronic functions, peptide synthesis by an enzyme method, and R and D on introduction of non-natural amino acid into peptides; in (2), element technologies were developed such as substrate forming technique (pattern forming and thin film forming technology), substrate ...

1994-03-01

448

Enhancement of Au nanoparticles formed by in situ electrodeposition on direct electrochemistry of myoglobin loaded into layer-by-layer films of chitosan and silica nanoparticles.

Extracellular ATP stimulates transmembrane ion fluxes in the mouse macrophage cell line J774. In the presence of Mg2+, nonhydrolyzable ATP analogs and other purine and pyrimidine nucleotides do not elicit this response, suggesting the presence of a specific receptor for ATP on the macrophage plasma membrane. One candidate for such a receptor is the ecto-ATPase expressed on these cells. We, therefore, investigated the role of this enzyme in ATP-induced /sup 86/Rb+ efflux in J774 cells. The ecto-ATPase had a broad nucleotide specificity and did not hydrolyze extracellular ATP in the absence of divalent cations. /sup 86/Rb+ efflux was not blocked by inhibition of the ecto-ATPase and did not require Ca2+ or Mg2+. In fact, ATP-stimulated /sup 86/Rb+ efflux was inhibited by Mg2+ and correlated with the availability of ATP4- in the medium. In the absence of divalent cations, the slowly hydrolyzable ATP analogs adenosine 5'-(beta, gamma-imido)triphosphate ...

1987-03-05

449

Energetics and kinetics of anaerobic aromatic and fatty acid degradation

In the present work, a new kind of myoglobin (Mb)/Au nanoparticles composite film was fabricated on pyrolytic graphite (PG) electrodes. Oppositely charged chitosan (CS) and silica (SiO(2)) nanoparticles were alternately adsorbed on the PG surface by the electrostatic interaction between them, forming {CS/SiO(2)}(5) layer-by-layer films. Mb and HAuCl(4) in solution were then simultaneously loaded into {CS/SiO(2)}(5) films. The loaded Au(III) in the films were electrochemically reduced into Au nanoparticles, forming nanocomposite films, designated as {CS/SiO(2)}(5)-Mb-Au. Various techniques such as cyclic voltammetry (CV), square wave voltammetry (SWV), quartz crystal microbalance (QCM), electrochemical impedance spectroscopy (EIS), scanning electron microscopy (SEM), and energy dispersive X-ray (EDX) analysis were used to characterize the films. Compared with {CS/SiO(2)}(5)-Mb films without Au nanoparticles inside, the {CS/SiO(2)}(5)-Mb-Au films exhibited much better behavior in ...

2008-12-01

450

Effects of chronic swimming training on cardiac sarcolemmal function and composition.

The kinetics of benzoate degradation by the anaerobic syntrophic bacterium, Syntrophus buswellii, was studied in coculture with Desulfovibrio strain G11. The threshold value for benzoate degradation was dependent on the acetate concentration with benzoate threshold values ranging from 2.4 [mu]M at 20 mM acetate to 30.0 [mu]M at 65 mM acetate. Increasing acetate concentrations also inhibited the rate of benzoate degradation with a apparent K[sub i] for acetate inhibition of 7.0 mM. Lower threshold values were obtained when nitrate rather than sulfate was the terminal electron acceptor. These data are consistent with a thermodynamic explanation for the threshold, and suggest that there is a minimum Gibbs free energy value required for the degradation of benzoate. An acetoacetyl-CoA thiolase has been isolated from Syntrophomonas wolfei; it is apparently a key enzyme controlling the synthesis of poly-B-hydroxyalkanoate from acetyl-CoA in this organism. Kinetic ...

1992-11-16

451

Effects of amifostine on radiation-induced apoptosis in mouse ovary

Cardiac contractile function is dependent on the integrity and function of the sarcolemmal membrane. Swimming exercise training is known to increase cardiac contractile performance. The purpose of the present study was to examine whether a swimming exercise program would alter sarcolemmal enzyme activity, ion flux, and composition in rat hearts. After approximately 11 wk of exercise training, cardiac myosin and actomyosin Ca2+-adenosinetriphosphatase (ATPase) activity was significantly higher in exercised rat hearts than in sedentary control rat hearts. Glycogen content was increased in plantaris and gastrocnemius muscles from exercised animals as was succinic dehydrogenase activity in gastrocnemius muscle of exercised rats in comparison to sedentary rat preparations. Sarcolemmal vesicles were isolated from hearts of exercise-trained and control rats. Sarcolemmal Na+-K+-ATPase and K+-p-nitrophenylphosphatase activities, Na+-Ca2+ exchange, and passive Ca2+ binding ...

1989-04-01

452

Detection of polychlorinated biphenyl degradation genes in polluted sediments by direct DNA extraction and polymerase chain reaction

The present study was designed to assess the radioprotective effects of amifostine on ovarian follicles. Three week-old female mice with or without pretreatment of amifostine were irradiated with 6.42 Gy of #gamma# -ray. Ovaries were collected 0 and 6h after irradiation. DNA fragmentation pattern and expression of genes and activity of proteins related with apoptosis were investigated by means of RT-PCR and Western blot. Proliferation of granulosa cells was reduced and incidence rate of follicular atresia was increased in ovarian follicles in #gamma# -ray irradiated mice compared to those in control or amifostine-treated group. DNA fragmentation was increased in time-dependent manner in granulosa cells of all irradiated groups. However, no difference between amifostine pre-treated group and irradiated groups was found and the expression of p53 as tumor suppressor gene and Bax as one of pro-apoptotic Bcl-2 family was increased in irradiated mice ovaries. PARP as DNA damage sensor was ...

2002-10-20

453

Deficiency of the housekeeping gene hypoxanthine-guanine phosphoribosyltransferase (HPRT) dysregulates neurogenesis.

It was the aim of this study to specifically detect the DNA sequences for the bphC gene, the meta-cleavage enzyme of the aerobic catabolic pathway for biphenyl and polychlorinated biphenyl degradation, in aquatic sediments without prior cultivation of microorganisms by using extraction of total DNA, PCR amplification of bphC sequences, and detection with specific gene probes. The direct DNA extraction protocol used was modified to enhance lysis efficiency. Crude extracts of DNA were further purified by gel filtration, which yielded DNA that could be used for the PCR. PCR primers were designed for conserved regions of the bphC gene from a sequence alignment of five known sequences. The specificity of PCR amplification was verified by using digoxigenin-labeled DNA probes which were located internal to the amplified gene sequence. The detection limit for the bphC gene of Pseudomonas paucimobilis Q1 and Pseudomaonas sp. strain LB 400-like sequences for the bphC gene ...

1993-12-01

454

Decreased duration of pentobarbital-induced narcosis in immature and adult female rats prenatally exposed to cimetidine

Neuronal transcription factors play vital roles in the specification and development of neurons, including dopaminergic (DA) neurons. Mutations in the gene encoding the purine biosynthetic enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) cause the resulting intractable and largely untreatable neurological impairment of Lesch-Nyhan disease (LND). The disorder is associated with a defect in basal ganglia DA pathways. The mechanisms connecting the purine metabolic defect and the central nervous system (CNS) phenotype are poorly understood but have been presumed to reflect a developmental defect of DA neurons. We have examined the effect of HPRT deficiency on the differentiation of neurons in the well-established human (NT2) embryonic carcinoma neurogenesis model. We have used a retrovirus expressing a small hairpin RNA (shRNA) to knock down HPRT gene expression and have examined the expression of a number of transcription factors essential for neuronal ...

2009-08-11

455

DNA rearrangements from #gamma#-irradiated normal human fibroblasts preferentially occur in transcribed regions of the genome

The effect of prenatal cimetidine exposure (PreCM) on the duration of pentobarbital-induced narcosis (DPN) was assessed in immature (14- and 28-day old) and adult (50-60-day old) male and female rats. PreCM exposure was accomplished by treating mothers with cimetidine (CM) (20 mg/kg, ip) daily for the last two days of gestation and then (0.01% in drinking water) throughout lactation. Pregnant mothers of untreated offspring (Con) received saline. PreCM decreased DPN to 505 +/- 33 min (from 611 +/- 23 min in Con) and 393 +/- 190 min (from 686 +/- 44 min in Con) in 14-day old male and female rats, respectively. Similarly, PreCM decreased DPN to 88 +/- 15 min (from 134 +/- 3 min in Con) and 102 +/- 19 min (from 171 +/- 44 min in Con) in 28-day old male and female rats, respectively. At 21 days, PreCM did not alter DPN in either sex. At 50-60 days, however, it decreased DPN to 144 +/- 41 min (from 238 +/- 7 min in Con) in females but had no effect in males; PreCM also increased the plasma ...

1986-03-01

456

Characterization of the transport of #alpha#-methylaminoisobutyric acid by a human intestinal cell line (HT-29)

Full text: DNA rearrangement events leading to chromosomal aberrations are central to ionizing radiation-induced cell death. Although DNA double-strand breaks are probably the lesion that initiates formation of chromosomal aberrations, little is understood about the molecular mechanisms that generate and modulate DNA rearrangement. Examination of the sequences that flank sites of DNA rearrangement may provide information regarding the processes and enzymes involved in rearrangement events. Accordingly, we developed a method using inverse PCR that allows the detection and sequencing of putative radiation-induced DNA rearrangements in defined regions of the human genome. The method can detect single copies of a rearrangement event that has occurred in a particular region of the genome and, therefore, DNA rearrangement detection does not require survival and continued multiplication of the affected cell. Ionizing radiation-induced DNA rearrangements were detected in ...

2003-08-17

457

Characterization of hyaluronate binding proteins isolated from 3T3 and murine sarcoma virus transformed 3T3 cells

Under certain growth conditions, the human colon adenocarcinoma cell line HT-29 exhibits intestinal enterocyte-like properties. The differentiated cells possess a brush border with the enzyme markers (aminopeptidase and sucrase) normally associated with the intestine. To aid in the characterization of the transport properties of these cells, the uptake of a non-metabolizable amino acid analog, "1"4C-#alpha#-methylaminoisobutyric acid (MeAIB) as examined in the HT-29-Al subclone which possesses a brush border. The cells exhibited a time-dependent uptake of MeAIB which was concentrative and sodium-dependent. The pH optimum for uptake was about 7.8. Uptake was inhibited by low temperature, 1 mM ouabain, or 0.5 mM dinitrophenol. A 1 hr-preincubation of the cells in an isotonic KCl solution resulted in a decreased uptake rate, suggesting that a negative membrane potential is important for MeAIB uptake. The rate of 0.5 mM MeABIB uptake was inhibited by 40 to 90% by 5 mM ...

1986-04-13

458

Biosynthesis of sucrose and mannitol as a function of leaf age in celery (Apium graveolens L. )

A hyaluronic acid binding fraction was purified from the supernatant media of both 3T3 and murine sarcoma virus (MSV) transformed 3T3 cultures by hyaluronate and immunoaffinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis resolved the hyaluronate affinity-purified fraction into three major protein bands of estimated molecular weight (M/sub r,e/) 70K, 66K, and 56K which contained hyaluronate binding activity and which were termed hyaluronate binding proteins (HABP). Hyaluronate affinity chromatography combined with immunoaffinity chromatography, using antibody directed against the larger HABP, allowed a 20-fold purification of HABP. Fractions isolated from 3T3 supernatant medium also contained additional binding molecules in the molecular weight range of 20K. This material was present in vanishingly small amounts and was not detected with a silver stain or with (/sup 35/S)methionine label. The three protein species isolated by hyaluronate affinity ...

1987-06-02

459

Basic fibroblast growth factor binds to subendothelial extracellular matrix and is released by heparitinase and heparin-like molecules

In celery (Apium graveolens L.), the two major translocated carbohydrates are sucrose and the acyclic polyol mannitol. Their metabolism, however, is different and their specific functions are uncertain. To compare their roles in carbon partitioning and sink-source transitions, developmental changes in /sup 14/CO/sub 2/ labeling, pool sizes, and key enzyme activities in leaf tissues were examined. The proportion of label in mannitol increased dramatically with leaf maturation whereas that in sucrose remained fairly constant. Mannitol content, however, was high in all leaves and sucrose content increased as leaves developed. Activities of mannose-6-P reductase, cytoplasmic and chloroplastic fructose-1,6-bis-phosphatases, sucrose phosphate synthase, and sucrose synthase increased with leaf maturation and decreased as leaves senesced. Ribulose bisphosphate carboxylase and nonreversible glyceraldehyde-3-P dehydrogenase activities rose as leaves developed but did not ...

1988-01-01

460

Assessment of phytoremediation as an in-situ technique for cleaning oil-contaminated sites

Basic fibroblast growth factor (bFGF) exhibits specific binding to the extracellular matrix (ECM) produced by cultured endothelial cells. Binding was saturable as a function both of time and of concentration of {sup 125}I-bFGF. Scatchard analysis of FGF binding revealed the presence of about 1.5 x 10{sup 12} binding sites/mm{sup 2} ECM with an apparent k{sub D} of 610 nM. FGF binds to heparan sulfate (HS) in ECM as evidenced by (i) inhibition of binding in the presence of heparin or HS at 0.1-1 {mu}g/mL, but not by chondroitin sulfate, keratan sulfate, or hyaluronic acid at 10 {mu}g/mL, (ii) lack of binding to ECM pretreated with heparitinase, but not with chondroitinase ABC, and (iii) rapid release of up to 90% of ECM-bound FGF by exposure to heparin, HS, or heparitinase, but not to chondroitin sulfate, keratan sulfate, hyaluronic acid, or chondroitinase ABC. Oligosaccharides derived from depolymerized heparin, and as small as the tetrasaccharide, released the ECM-bound FGF, but there ...

1989-02-21

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461

An Arabidopsis thaliana methyltransferase Capable of Methylating Farnesoic Acid

Literature on examples of phytoremediation techniques used in the in-situ remediation of soils contaminated by petroleum hydrocarbons is reviewed. The review includes discussion of the key mechanisms involved in each case, benefits, limitations and costs compared to alternative approaches, including natural attenuation, engineering and bioremediation. Review of the literature led to the conclusion that phytoremediation is an effective method for degrading and containing petroleum hydrocarbons in soil, and confirmed the ability of plants to transfer volatile petroleum hydrocarbons, such as napthalene, from the soil to the atmosphere via transpiration. The primary loss mechanism for the degradation of petroleum hydrocarbons appears to be microorganisms in the rhizosphere of plants. The available information also suggests that plants may degrade petroleum hydrocarbons directly, although the indirect role played by plants is far more common. These roles include supplying root exudates for ...

462

Aflatoxin-producing Aspergillus spp. and aflatoxin levels in stored cassava chips as affected by processing practices

We previously reported the identification of a new family of plant methyltransferases (MTs), named the SABATH family, that use S-adenosyl-l-methionine (SAM) to methylate a carboxyl moiety or a nitrogen-containing functional group on a diverse array of plant compounds. The Arabidopsis genome alone contains 24 distinct SABATH genes. To identify the catalytic specificities of members of this protein family in Arabidopsis, we screened recombinantly expressed and purified enzymes with a large number of potential substrates. Here, we report that the Arabidopsis thaliana gene At3g44860 encodes a protein with high catalytic specificity towards farnesoic acid (FA). Under steady-state conditions, this farnesoic acid carboxyl methyltransferase (FAMT) exhibits K{sub M} values of 41 and 71 {mu}M for FA and SAM, respectively. A three-dimensional model of FAMT constructed based upon similarity to the experimentally determined structure of Clarkia breweri salicylic acid ...

2006-01-01

463

Activation of rapid signaling pathways and the subsequent transcriptional regulation for the proliferation of breast cancer MCF-7 cells by the treatment with an extract of Glycyrrhiza glabra root.

Cassava chips (cassava balls, and cassava pellets) are derived cassava products traditionally produced by farmers in sub-Saharan Africa following fermentation, and drying of fresh roots of cassava, and are widely consumed in Cameroon. Once produced, this food commodity can be stored for more than two months and contaminated by a wide array of harmful microbes. In order to assess persistence of toxigenic fungi in cassava chips, aflatoxin-producing fungi (Aspergillus flavus, Aspergillus nomius, and Aspergillus parasiticus) and aflatoxins were contrasted at regular intervals in home-stored cassava chips collected in two locations of southern Cameroon throughout a two-month monitoring period. Three hundred and forty-six isolates of aflatoxin-producing fungi were found to be associated with all samples. A. flavus contaminated more samples in both types of chips (267 isolates in 53 samples), followed by A. nomius (58 isolates in 15 samples), whereas A. parasiticus was rarest. A direct ...

2009-01-01

464

A single amino acid substitution (R441A) in the receptor-binding domain of SARS coronavirus spike protein disrupts the antigenic structure and binding activity

Glycyrrhiza glabra root is one of the common traditional Chinese medicines and used as flavoring and sweetening agents for tobaccos, chewing gums, candies, toothpaste and beverages. While glycyrrhizin is one of the main components in the extract of G. glabra root and has been characterized, the other components have not been well characterized. The mechanism of growth activation of breast cancer MCF-7 cells, including the activation of Erk1/2 and Akt, and the transcriptional regulation of estrogen-responsive genes, was examined by means of sulforhodamine B, luciferase reporter gene, real-time RT-PCR and Western blotting assays after the induction of the cells with the extract of G. glabra root. The extract has similar activity to that induced by 17beta-estradiol (E(2)), although glycyrrhizin did not show such an activity. Moreover, the estrogen receptor alpha-dependent neurite outgrowth induced by the extract was similar to that by E(2), whereas glycyrrhizin had no effect. Furthermore, ...

2007-06-24

465

A criterion to determine whether cis-4-hydroxyproline is produced in animal tissues

The spike (S) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) has two major functions: interacting with the receptor to mediate virus entry and inducing protective immunity. Coincidently, the receptor-binding domain (RBD, residues 318-510) of SAR-CoV S protein is a major antigenic site to induce neutralizing antibodies. Here, we used RBD-Fc, a fusion protein containing the RBD and human IgG1 Fc, as a model in the studies and found that a single amino acid substitution in the RBD (R441A) could abolish the immunogenicity of RBD to induce neutralizing antibodies in immunized mice and rabbits. With a panel of anti-RBD mAbs as probes, we observed that R441A substitution was able to disrupt the majority of neutralizing epitopes in the RBD, suggesting that this residue is critical for the antigenic structure responsible for inducing protective immune responses. We also demonstrated that the RBD-Fc bearing R441A mutation could not bind to soluble and cell-associated ...

2006-05-26

466

A criterion to determine whether cis-4-hydroxyproline is produced in animal tissues

Hydrolyzates of tissues that had been labeled with (14C)proline often contain significant amounts of cis-4-hydroxy(14C)proline. Since animal cells do not contain an enzyme which can effect formation of cis-4-hydroxyproline, there are only two possible explanations for its presence. Either it is formed during acid hydrolysis of trans-4-hydroxyproline (which is synthesized by cells and is a common constituent of connective tissues), or it is produced by a nonenzymatic mechanism such as attack by oxygen radicals. It is important to resolve this issue because if a nonenzymatic mechanism is active in connective tissues, then it will be necessary to reevaluate currently accepted ideas about production of hydroxyproline. This communication describes a method for distinguishing between the two alternate explanations. Tissues or cells are labeled with (14C)proline, and then a known amount of trans-4-hydroxy(3H)proline is added to each sample before hydrolysis; the relative ...

1984-03-01

467

Protective role of selenium against renal toxicity induced by cadmium in rats

Hydrolyzates of tissues that had been labeled with [14C]proline often contain significant amounts of cis-4-hydroxy[14C]proline. Since animal cells do not contain an enzyme which can effect formation of cis-4-hydroxyproline, there are only two possible explanations for its presence. Either it is formed during acid hydrolysis of trans-4-hydroxyproline (which is synthesized by cells and is a common constituent of connective tissues), or it is produced by a nonenzymatic mechanism such as attack by oxygen radicals. It is important to resolve this issue because if a nonenzymatic mechanism is active in connective tissues, then it will be necessary to reevaluate currently accepted ideas about production of hydroxyproline. This communication describes a method for distinguishing between the two alternate explanations. Tissues or cells are labeled with [14C]proline, and then a known amount of trans-4-hydroxy[3H]proline is added to each sample before hydrolysis; the relative ...

1984-01-01

468

Prostaglandin synthesis by chicken and rat lung microsomes

Cadmium is an environmental toxic metal implicated in human diseases. The mechanism of its toxicity is not fully understood. Therefore, the role of cadmium in renal toxicity, and the protective role of selenium against this toxicity were investigated. Forty-five male rats were used through out the study and divided into three groups of 15. The first group received saline solution daily for 10 days. The second group, received cadmium chloride (CdCl_2) (2 mg/kg body weight) intraperitoneally daily for a period of 10 days. The third group, received sodium selenite (1 mg/kg body weight, twice a day) and CdCl_2 (once a day) for a period of 10 days. The results showed that cadmium treatment increased renal lipid peroxidation (measured as malondialdehyde, MDA) which was associated with a significant decrease in the antioxidant systems such as reduced glutathione levels and the activities of glutathione peroxidase (GPx) and thioredoxin reductase (TrxR). On the other hand, pretreatment of rats ...

2007-06-25

469

Genes involved in immortalization of human mammary cells

A comparison between chicken and rat lung was made for microsomal prostaglandin (PG) synthesis from 1-/sup 14/C-arachidonic acid. Microsomal protein (2.0 mg) from chicken or rat lung was incubated in the presence of 20 ..mu..g of 1-/sup 14/C-arachidonic acid (specific activity = 3 x 10/sup 6/ dpm/..mu..mol for chicken; 6 x 10/sup 6/ dpm/..mu..mol for rat), 0.05 M Tris-HCl buffer (pH = 8.0), 0.5 mM epinephrine, and 1 mM reduced glutathione in a total volume of 0.5 ml in a 37/sup 0/C water bath with shaking for 15 min. After acidification with 1 M HCl to pH 3, prostaglandins were extracted with ethyl acetate. The products of the reactions were separated by reversed phase chromatography, and the radioactivity of each prostanoid fraction was determined. The predominant prostanoid synthesized by chicken lung microsomes was PGE/sub 2/, followed by much lower amounts of thromboxane B/sub 2/ (TXB/sub 2/), PGF/sub 2//sub ..cap alpha../ and PGD/sub 2/. In at lung, 6-keto-PGF/sub 1//sub ..cap ...

1986-03-01

470

Development of detection methods for irradiated foods; development of immunological identification of irradiated foods

Breast cancer progression is characterized by inappropriate cell growth. Normal cells cease growth after a limited number of cell divisions--a process called cellular senescence-while tumor cells may acquire the ability to proliferate indefinitely (immortality). Inappropriate expression of specific oncogenes in a key cellular signaling pathway (Ras, Raf) can promote tumorigenicity in immortal cells, while causing finite lifespan cells to undergo a rapid senescence-like arrest. We have studied when in the course of transformation of cultured human mammary epithelial cells (HMEC), the response to overexpressed oncogenic Raf changes from being tumor-suppressive to tumor enhancing, and what are the molecular underpinnings of this response. Our data indicate: (1) HMEC acquire the ability to maintain growth in the presence of oncogenic Raf not simply as a consequence of overcoming senescence, but as a result of a newly discovered step in the process of immortal transformation uncovered by ...

2001-09-27

471

An ELISA-based high throughput protein truncation test for inherited breast cancer

Enzyme-linked Immunosorbent assay systems for the identification of irradiated egg, pork and chicken was developed. Eggs were irradiated in their shells to 0.5{approx}7kGy. Pork was irradiated to 0.5{approx}3kGy and chicken irradiated to 0.5kGy{approx}5kGy. The most sensitive proteins to irradiation were screened by SDS-PAGE and purified. Ovalbumin from egg, salt soluble protein(p) from pork, and salt soluble protein(c) from chicken showed the most sensitivity to irradiation. To investigate for a practical use in identifying of irradiated egg, pork and chicken, competitive ELISA was performed. The binding activity of ovalbumin to anti-ovalbumin IgG was reduced in a dose-dependent manner by irradiating up to 7kGy, and considerably lowered after irradiating at 7kGy. The concentration of 50% inhibition of ovalbumin to IgG was increased to 1.5(0.5kGy){approx}3.7(7kGy) times in an dose-dependent relationship. The binding activity of salt soluble protein(p) to anti-salt ...

2002-04-01

472

The biodistribution and effect on hepatic parenchyma with intraarterial injected I-131 lipiodol into hepatic artery

IntroductionBreast cancer is the most diagnosed and second leading cause of cancer deaths in the U.S. female population. An estimated 5 to 10 percent of all breast cancers are inherited, caused by mutations in the breast cancer susceptibility genes (BRCA1/2). As many as 90% of all mutations are nonsense mutations, causing a truncated polypeptide product. A popular and low cost method of mutation detection has been the protein truncation test (PTT), where target regions of BRCA1/2 are PCR amplified, transcribed/translated in a cell-free protein synthesis system and analyzed for truncated polypeptides by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography. We previously reported a novel High Throughput Solid-Phase PTT (HTS-PTT) based on an enzyme-linked immunosorbent assay (ELISA) format that eliminates the need for radioactivity, SDS-PAGE and subjective interpretation of the results. Here, we report the next generation HTS-PTT ...

2010-10-04

473

Contamination by depleted uranium (Du) in South Serbia

Iodized oil has been used as a contrast agent in lymphangiography. One of the commercially available compounds is Lipidol Ultra-fluid(LUF) which contains 38% iodine by weight. Nakakuma et al(1979) reported that LUF was selectively retained in the hypervascular hepatocellular carcinoma when injected directly into the ligated hepatic artery. Since that time, it has been widely utilized in the detection as well as the therapeutic attempts of hepatocellular carcinoma, where it has been mixed with chemotherapeutic agents or labeled with radioactive I-131. Like all significant advances, the mechanism of lipid retention within the hepatocellular carcinoma is not clearly understood, and also there is a lack of information about the biodistribution and kinetics of I-131 Lipiodol. The apparent safety of this technique require confirmation. The present study was aimed to assess the biodistribution and kinetics of intraarterially injected I-131 Lipiodol and the histologic changes in canine livers. ...

1989-08-15

474

Yield-increasing additives in kraft pulping: Effect on carbohydrate retention, composition and handsheet properties

The paper present the results of the study on D.U. (depleted uranium) contamination in the environment and possible effects on animal healths in the region o f Bujanovac. Samples of soil, feed, leaves, grass, lichen, moss, honey and water were collected randomly in 2003/2004 in the vicinity of the target area (500-1000 m) and 5 km from the target area. Activity of the radionuclides ({sup 226}Ra, {sup 232}Th, {sup 40}K, {sup 210}Pb, {sup 238}U, {sup 235}U, {sup 137}Cs, {sup 7}Be) in soils, grass, lichen, moss and honey was determined on Hp Ge detector (Canberra, relative efficiency 23%) by standard gamma spectrometry. Total alpha and beta activity in water was determined on proportional alpha/beta counter (Canberra 2400, efficiency for alpha emitters 11%, efficiency for beta emitters 30%). Non significantly higher values of concentrations of {sup 226}Ra, {sup 232}Th, {sup 238}U and {sup 235}U were measured in the immediate vicinity of the targeted site, but {sup 235}U/{sup 238}U ...

2006-07-01

475

Breaking the Biological barriers to Cellulosic Ethanol: A Joint Research Agenda

In this thesis, increased hemicellulose retention during kraft pulping has been studied. The work has been divided into three parts: i) Development of an accessible and reliable method for determination of carbohydrate composition of kraft pulps ii) Investigation of the composition and molecular mass distributions of the carbohydrates in kraft pulps with increased hemicellulose content iii) Investigation of the effect of increased hemicellulose content on the sheet properties of kraft pulps with increased hemicellulose content. A method for carbohydrate determination was developed. In this method, enzymes are used to hydrolyse the pulp into monosaccharides. A relatively mild acid hydrolysis is performed prior to detection on an HPLC with an RI-detector. The pulp is not derivatized and no pre-treatment (mechanical or chemical) is needed to determine the carbohydrate composition using the method developed here. Peak deconvolution software is used to improve the ...

2008-07-01

476