yeast atp synthase: Topics by WorldWideScience.org

Two rotary motors in F-ATP synthase are elastically coupled by a flexible rotor and a stiff stator stalk

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

Double-lock ratchet mechanism revealing the role of ?SER-344 in F_oF₁ ATP synthase

ATP is synthesized by ATP synthase (F_OF₁-ATPase). Its rotary electromotor (F_O) translocates protons (in some organisms sodium cations) and generates torque to drive...Full Text Available

2011-03-08

Phosphorylation of sites 3 and 2 in rabbit skeletal muscle glycogen synthase by a multifunctional protein kinase (ATP-citrate lyase kinase)

In a majority of living organisms, F_oF₁ ATP synthase performs the fundamental process of ATP synthesis. Despite the simple net reaction formula, ADP + P_i → ATP + H₂O,...Full Text Available

2011-03-22

Identification and Characterization of CPS1 as a Hyaluronic Acid Synthase Contributing to the Pathogenesis of Cryptococcus neoformans Infection?

A multifunctional protein kinase, purified from rat liver as ATP-citrate lyase kinase, has been identified as a glycogen synthase kinase. This kinase catalyzed incorporation of up to 1.5 mol of and)2numberSPO4/mol of synthase subunit associated with a decrease in the glycogen synthase activity ratio from 0.85 to a value of 0.15. Approximately 65-70% of the TUPO4 was incorporated into site 3 and 30-35% into site 2 as determined by reverse phase high performance liquid chromatography. This multifunctional kinase was distinguished from glycogen synthase kinase-3 on the basis of nucleotide and protein substrate specificities. Since the phosphate contents in glycogen synthase of sites 3 and 2 are altered in diabetes and by insulin administration, the possible involvement of the multifunctional kinase was explored. Glycogen synthase purified from ...

1985-10-05

Activation and Stiffness of the Inhibited States of F₁-ATPase Probed by Single-molecule Manipulation

Cryptococcus neoformans is a pathogenic yeast that often causes devastating meningoencephalitis in immunocompromised individuals. We have previously identified the C. neoformans...Full Text Available

2007-08-01

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

F₁-ATPase (F₁), a soluble portion of F_oF₁-ATP synthase (F_oF₁), is an ATP-driven motor in which γϵ subunits rotate in...Full Text Available

2010-04-09

Regulation of starch synthesis in potato tubers

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 FAD-binding dinucleotide folds. The active sites ...

1989-11-01

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

Following tuber excision from the mother plant sucrose synthase activity fell from 3,120 to 960 nmol/g.f. wt./h within 7 days and starch synthesis ({sup 14}C sucrose incorporated into isolated discs) from 23 to 7 nmol/g.f. wt./h. While the maximum catalytic activity of sucrose synthase was more than sufficient to account for the observed rate of starch synthesis a maximum of 27% of sucrose incorporated by discs was converted into starch within 3 h. This compared with 80% conversion of {sup 14}C glucose incorporated. Tuber excision also reduced the rate of starch biosynthesis with glucose as a substrate (from 206 to 64 nmol/g.f. wt./h). The activities of UDPG-pyrophosphorylase, PPi-PFK, ATP-PFK, starch synthase and hexokinase (glucose or fructose substrates) were unaffected by tuber removal. ADPG pyrophosphorylase activity was reduced from 8,000 to 4,500 nmol/g.f. wt./h. Preliminary experiments indicate ...

1990-05-01

Vascular ATP-sensitive potassium channels are over-expressed and partially regulated by nitric oxide in experimental septic shock

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 ...

1997-12-01

British Library Electronic Table of Contents (United Kingdom)

Purpose To study the activation and expression of vascular (aorta and small mesenteric arteries) potassium channels during septic shock with or without modulation of the NO pathway. Methods Septic shock was induced in rats by peritonitis. Selective inhibitors of vascular KATP (PNU-37883A) or BKCa [iberiotoxin (IbTX)] channels were used to demonstrate their involvement in vascular hyporeactivity. Vascular response to phenylephrine was measured on aorta and small mesenteric arteries mounted on a wire myograph. Vascular expression of potassium channels was studied by PCR and Western blot, in the presence or absence of 1400W, an inducible NO synthase (iNOS) inhibitor. Aortic activation of the transcriptional factor nuclear factor-kappaB (NF-?B) was assessed by electrophoretic mobility shift as...

2011-01-01

Photolabelling kinetics of ATP-ase of sarcoplasmatic reticulum by 3'-ARILAZIDO-ATP

Supramolecular Structures for Photochemical Energy Conversion

... atp-ase azido compounds biochemical reaction kinetics endoplasmic reticulum

1984-05-02

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

OAK B188 The goal of this project is to mimic the energy transduction processes by which photosynthetic organisms harvest sunlight and convert it to forms of energy that are more easily used and stored. The results may lead to new technologies for solar energy harvesting based on the natural photosynthetic process. They may also enrich our understanding and control of photosynthesis in living organisms, and lead to methods for increasing natural biomass production, carbon dioxide removal, and oxygen generation. In our work to date, we have learned how to make synthetic antenna and reaction center molecules that absorb light and undergo photoinduced electron transfer to generate long-lived, energetic charge-separated states. We have assembled a prototype system in which artificial reaction centers are inserted into liposomes (artificial cell-like constructs), where they carry out light-driven transmembrane translocation of hydrogen ions to generate proton motive force. By insertion of ...

2003-08-26

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

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. ...

1991-05-01

Molecular Cloning and Characterization of the Yew Gene Encoding Squalene Synthase from Taxus cuspidata

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 ...

1987-03-05

Immobilization on Chitosan of a Thermophilic Trehalose Synthase from Thermus thermophilus HJ6

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2007-09-01

Cell Cycle-dependent Expression of Chitin Synthase Genes in Aspergillus nidulans

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2010-03-01

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

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2001-03-01

The ATP-Binding Cassette Transporter ABCA4: Structural and Functional Properties and Role in Retinal Disease

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 ...

ATP overflow in skeletal muscle 1A arterioles

ATP-binding cassette transporters (ABC transporters) utilize the energy of ATP hydrolysis to translocate an unusually diverse set of substrates across cellular membranes. ABCA4, also known as...Full Text Available

2010-01-01

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Survival of Genetically Modified and Self-Cloned Strains of Commercial Baker's Yeast in Simulated Natural Environments: Environmental Risk Assessment

The purpose of this study was to investigate the sources of ATP in the 1A arteriole, and to investigate age-related changes in ATP overflow. Arterioles (1A) from the red portion of the gastrocnemius...Full Text Available

2010-08-15

Comparison of the Survival and Metabolic Activity of Psychrophilic and Mesophilic Yeasts Subjected to Freeze-Thaw Stress

Although genetic engineering techniques for baker's yeast might improve the yeast's fermentation characteristics, the lack of scientific data on the survival of such strains in natural environments...Full Text Available

2005-11-01

Global Transcriptional Responses of Fission Yeast to Environmental Stress

A mesophilic yeast, Candida utilis, and a psychrophilic yeast, Leucosporidium stokesii, were subjected to freeze-thaw cycling over the range 25 to -60 C. Viability...Full Text Available

1975-06-01

Apoptosis at Inflection Point in Liquid Culture of Budding Yeasts

We explored transcriptional responses of the fission yeast Schizosaccharomyces pombe to various environmental stresses. DNA microarrays were used to characterize changes in expression...Full Text Available

2003-01-01

ATP-gated P2X₃ receptors constitute a positive autocrine signal for insulin release in the human pancreatic ? cell

Budding yeasts are highly suitable for aging studies, because the number of bud scars (stage) proportionally correlates with age. Its maximum stages are known ...Full Text Available

Structure and Reaction Mechanism of Basil Eugenol Synthase

Extracellular ATP has been proposed as a paracrine signal in rodent islets, but it is unclear what role ATP plays in human islets. We now show the presence of an ATP signaling pathway that enhances...Full Text Available

2010-04-06

Partial Purification of Digitonin-Solubilized ?-Glucan Synthase from Red Beet Root ¹

Phenylpropenes, a large group of plant volatile compounds that serve in multiple roles in defense and pollinator attraction, contain a propenyl side chain. Eugenol synthase (EGS) catalyzes the reductive...Full Text Available

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

An enriched glucan synthase fraction was obtained from red beet root microsomes by sequential extraction with the detergents 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate and digitonin....Full Text Available

1987-04-01

Expression in E. coli and characterization of the catalytic domain of Botrytis cinerea chitin synthase

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

Differentially regulated malate synthase genes participate in carbon and nitrogen metabolism of S. cerevisiae.

BackgroundChitin synthase 3a (CHS3a) from Botrytis cinerea (Bc) catalyses the multiple transfer of N-acetylglucosamine (GlcNAc) residues to the...Full Text Available

Differential expression of prostaglandin-H synthase isoenzymes in normal and activated keratinocytes in vivo and in vitro.

We have isolated a second gene (MLS1), which in addition to DAL7, encodes malate synthase from S. cerevisiae. Expression of the two genes is specific for their physiological roles in carbon and nitrogen...Full Text Available

1992-11-11

Dexamethasone and nitric oxide synthase gene expression in brain.

Normal mouse epidermis constitutively expresses prostaglandin-H synthase 1 (PGHS-1) but no PGHS-2. Acute inflammation and epidermal hyperplasia, (hyperplastic transformation), as evoked in adult mouse...Full Text Available

1995-07-01

Cottonseed Malate Synthase ¹

Systemic administration of lipopolysaccharide (LPS), which causes endotoxemia and systemic inflammation, has been reported to induce expression of the gene for type II inducible nitric oxide synthase...Full Text Available

1997-03-01

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

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

Carbon Partitioning in Eelgrass (Regulation by Photosynthesis and the Response to Daily Light-Dark Cycles).

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

Acetolactate Synthase Activity in Developing Maize (Zea mays L.) Kernels

Diel variations in rates of C export, sucrose-phosphate synthase (SPS) and sucrose synthase (SS) activity, and C reserves were investigated in Zostera marina L. (eelgrass) to elucidate the environmental...Full Text Available

1995-08-01

Promoter Structure Which Affects on the Expression of Yeast MGMT Gene

Acetolactate synthase (EC 4.1.3.18) activity was examined in maize (Zea mays L.) endosperm and embryos as a function of kernel development. When assayed using unpurified homogenates,...Full Text Available

1988-01-01

Effect of Zinc-Enriched Yeast Supplementation on Serum Zinc and Testosterone Concentrations in Ethanol Feeding Rats

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1997-01-01

Application of Quantitative Real-Time PCR for Enumeration of Total Bacterial, Archaeal, and Yeast Populations in Kimchi

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2008-07-01

Terpenoid-based defenses in conifers: cDNA cloning, characterization, and functional expression of wound-inducible (E)-?-bisabolene synthase from grand fir (Abies grandis)

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2009-12-01

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Contraction-mediated inactivation of glycogen synthase is accompanied by inactivation of glycogen synthase phosphatase in human skeletal muscle.

(E)-α-Bisabolene synthase is one of two wound-inducible sesquiterpene synthases of grand fir (Abies grandis), and the olefin product of this cyclization reaction...Full Text Available

1998-06-09

Mechanism of adenylate kinase. Dose adenosine 5'-triphosphate bind to the adenosine 5'-monophosphate site

Activities of glycogen synthase (GS) and GS phosphatase were determined on human muscle biopsies before and after isometric contraction at 2/3 maximal voluntary force. Total GS activity did not change...Full Text Available

1989-05-01

Matching ATP Supply and Demand in Mammalian Heart: In Vivo, In Vitro and In Silico Perspectives

Although the subtrate binding properties of adenylate kinase (AK) have been studied extensively by various biochemical and biophysical techniques, it remains controversial whether uncomplexed adenosine 5'-triphosphate (ATP) binds to the adenosine 5'-monophosphate (AMP) site of AK. The authors present two sets of experiments which argue against binding of ATP to the AMP site. (a) /sup 31/P nuclear magnetic resonance titration of ATP with AK indicated a 1:1 stoichiometry on the basis of changes in coupling constants and line widths. This ruled out binding of ATP to both sites. (b) ATP and MgATP were found to behave similarly by protecting AK from spontaneous inactivation while AMP showed only a small degree of protection. Such inactivation could also be protected or reversed by dithioerythritol and is most likely due to oxidation of sulfhydryl ...

1987-10-06

Determination of Adenosine Triphosphate on Marine Particulates:Synthesis of Methods for Use on OTEC Samples

Although the heart rapidly adapts cardiac output to match the body’s circulatory demands, the regulatory mechanisms ensuring that sufficient ATP is available to perform the required...Full Text Available

2010-02-01

Modular coherence of protein dynamics in yeast cell polarity system

Adenosine triphosphate (ATP) is an indicator of living biomass in marine particulates. This report details the method used by Lawrence Berkeley Laboratory to analyze particulate ATP in samples taken from oligotrophic, tropical ocean waters. It represents a synthesis of previously published methods.

1982-08-01

METABOLIC REGULATION OF ADENOSINE TRIPHOSPHATE SULFURYLASE IN YEAST

In this study, we investigated on a systems level how complex protein interactions underlying cell polarity in yeast determine the dynamic association of proteins with the polar cortical domain (PCD)...Full Text Available

2011-05-03

In vivo expression and mitochondrial targeting of yeast apoiso-1-cytochrome c fusion proteins.

de Vito, Peter C. (Princeton University, Princeton, N.J.), and Jacques Dreyfuss. Metabolic regulation of adenosine triphosphate sulfurylase in yeast. J. Bacteriol. 88:1341–1348....Full Text Available

1964-11-01

Heavy metals alter the electrokinetic properties of bacteria, yeasts, and clay minerals.

To define the import pathway for apoiso-1-cytochrome c in vivo, the coding region for bacterial chloramphenicol acetyltransferase (CAT) or yeast copper metallothionein (CuMT) was fused to the carboxy...Full Text Available

1990-11-01

Evaluation of Acyl Coenzyme A Oxidase (Aox) Isozyme Function in the n-Alkane-Assimilating Yeast Yarrowia lipolytica

The electrokinetic patterns of four bacterial species (Bacillus subtilis, Bacillus megaterium, Pseudomonas aeruginosa, and Agrobacterium radiobacter), two yeasts (Saccharomyces cerevisiae and Candida...Full Text Available

1992-05-01

Antagonistic Gcn5-Hda1 interactions revealed by mutations to the Anaphase Promoting Complex in yeast

We have identified five acyl coenzyme A (CoA) oxidase isozymes (Aox1 through Aox5) in the n-alkane-assimilating yeast Yarrowia lipolytica, encoded by the POX1...Full Text Available

1999-09-01

A Novel Function of the DNA Repair Gene rhp6 in Mating-Type Silencing by Chromatin Remodeling in Fission Yeast

BackgroundHistone post-translational modifications are critical for gene expression and cell viability. A broad spectrum of histone lysine residues have been identified in yeast...Full Text Available

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

Recent studies have indicated that the DNA replication machinery is coupled to silencing of mating-type loci in the budding yeast Saccharomyces cerevisiae, and a similar silencing mechanism...Full Text Available

1998-09-01

DNA double strand breakage after UV irradiation of yeast cells; DNA-Doppelstrangbrueche nach UV-Bestrahlung in Hefezellen

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 ...

1986-06-08

Cholera toxin binding sites in yeast triggers biochemical pathway

No abstract prepared.

2000-07-01

Characterization of commercially available selenized yeast food supplements

... e Biologia Molecular (SBBq), Sao Paulo, SP (Brazil) 217 p. APPLIED LIFE

1998-05-23

Effect of UV radiation on the killer phenotype in the wine yeast-saccharomycetes and spontaneous variation of this character

... Rome (Italy) Istituto Superiore di Sanita (Italy) 207 p. ENVIRONMENTAL

2009-04-01

Biosynthesis of hyaluronan: direction of chain elongation

Spontaneous and ultraviolet-induced changeabilities of wine yeasts from the killer state to sensitive one have been studied. Observed often spontaneous changes of killer and neutral phenotypes under laboratory store conditions as well as high mutation frequency of genetic elements responsible for the killer indication on ultraviolet irradiation testify that often encounterability in nature and in the production of sensitive yeasts is attributed to high frequency of mutation changes of the killer and neutral phenotypes to the sensitive state.

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

The mechanism of hyaluronan biosynthesis in vertebrates had been proposed to occur at the reducing end of growing chains. This mechanism was questioned because a recombinant synthase appeared to add...Full Text Available

2006-09-15

Thio Wax Ester Biosynthesis Utilizing the Unspecific Bifunctional Wax Ester Synthase/Acyl Coenzyme A:Diacylglycerol Acyltransferase of Acinetobacter sp. Strain ADP1

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

Pharmacokinetics and pharmacodynamics of terbogrel, a combined thromboxane A₂ receptor and synthase inhibitor, in healthy subjects

The bifunctional wax ester synthase/acyl coenzyme A (acyl-CoA):diacylglycerol acyltransferase (WS/DGAT) from Acinetobacter sp. strain ADP1 (formerly Acinetobacter calcoaceticus...Full Text Available

2005-02-01

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Campylobacter jejuni Fatty Acid Synthase II: Structural and functional analysis of ?-hydroxyacyl-ACP dehydratase (FabZ)

AimsTo characterize the pharmacokinetics of terbogrel, a new combined thromboxane A₂ (TxA₂) receptor and synthase inhibitor, in healthy human subjects after...Full Text Available

2004-07-01

Biological effects of exogenous adenosine 5 prime -triphosphate on cultured mammalian cells: Evidence for a receptor mechanism and its regulation by desensitization

Fatty acid biosynthesis is crucial for all living cells. In contrast to higher organisms, bacteria use a type II fatty acid synthase (FAS II) composed of a series of individual proteins, making...Full Text Available

2009-03-06

Adenosine triphosphate loading thallium-201 myocardial scintigraphy. Optimal dose and diagnostic accuracy

Exogenous adenosine 5{prime}-triphosphate (ATP) mobilized intracellular calcium in human carcinoma A43l cells and in Swiss 3T3 and 3T6 mouse fibroblasts by increasing inositol trisphosphate similar to well down growth factors (platelet-derived growth factor (PDGF), epidermal growth factor (EGF), bradykinin (BK), serum). Calcium mobilization was examined by video imaging of fura-2 fluorescence is single cells, following the radioactive isotope {sup 45}Ca, and monitoring the decrease in fluorescence of cells loaded with chlortetracycline. Uridine 5{prime}-triphosphate, but not other nucleotides, mimicked ATP. Single-cell analysis revealed synchronous responses in 10 sec to ATP, BK or serum, while PDGF (3T3) and EGF (A431) produced slower signals with significant cell-to-cell variation. PDGF desensitized 3T3 cells to ATP and BK added 100 sec later but ATP or BK did not desensitized to ...

1989-01-01

C-106 tank process ventilation test

Adenosine triphosphate (ATP) is an alternative to dipyridamole or adenosine in thallium-201 myocardial scintigraphy. However, the optimal dose of ATP has not been determined. A Doppler guide wire study showed the coronary flow velocity at a dose of 0.15 mg/kg of ATP was equal or higher than that at 0.14 mg/kg of adenosine or 0.56 mg/kg of dipyridamole. ATP was given intravenously to 67 patients with coronary artery disease at 0.15 mg/kg/min for 6 min. Thallium-201 was injected at 3 min, followed by immediate and delayed (3 hrs) tomographic imaging. There was no serious side effect during examination, although chest pain (26%), dyspnea (17%), and flushing (33%) were common. The sensitivity and specificity to detect coronary artery disease were 98 and 100%, respectively. The sensitivity to detect left anterior descending artery, left circumflex artery, and right coronary artery lesions was 94, 59 and 77%, ...

1995-01-01

Some ABCA3 mutations elevate ER stress and initiate apoptosis of lung epithelial cells

Project W-320 Acceptance Test Report for tank 241-C-106, 296-C-006 Ventilation System Acceptance Test Procedure (ATP) HNF-SD-W320-012, C-106 Tank Process Ventilation Test, was an in depth test of the 296-C-006 ventilation system and ventilation support systems required to perform the sluicing of tank C-106. Systems involved included electrical, instrumentation, chiller and HVAC. Tests began at component level, moved to loop level, up to system level and finally to an integrated systems level test. One criteria was to perform the test with the least amount of risk from a radioactive contamination potential stand point. To accomplish this a temporary configuration was designed that would simulate operation of the systems, without being connected directly to the waste tank air space. This was done by blanking off ducting to the tank and connecting temporary ducting and an inlet air filter and housing to the recirculation system. This configuration would eventually ...

1998-07-20

Radiosensitization of human lung fibroblasts by chemicals that decrease ATP levels

BackgroundABCA3 transporter (ATP-binding cassette transporter of the A subfamily) is localized to the limiting...Full Text Available

2011-01-01

P2Y receptors on astrocytes and microglia mediate opposite effects in astroglial proliferation

Radiosensitization by lactate, pyruvate, nalidixic acid and novobiocin was studied in exponentially growing SH-18L human lung fibroblasts. All the chemicals had a slight radiosensitizing effect at a low concentration and a definite effect at a higher one. Decreases in the D_0 and/or Dq values were present in each dose survival curve. Fibroblasts incubated with the low concentration of each chemical for 24 hrs after X irradiation showed no reduction in intracellular ATP content, whereas, the higher concentration produced a significant decrease. These observations suggest that the decrease in the ATP content may be involved in the radiosensitization of human fibroblasts at high concentrations of these chemicals. In contrast, radiosensitization at a low concentration is not explained by a relationship to ATP content. Different mechanisms may be involved in radiosensitization at low and high concentrations of these chemicals. ...

British Library Electronic Table of Contents (United Kingdom)

Nucleotides released upon brain injury signal to astrocytes and microglia playing an important role in astrogliosis, but the participation of microglia in the purinergic modulation of astrogliosis is still unclear. Highly enriched astroglial cultures and co-cultures of astrocytes and microglia were used to investigate the influence of microglia in the modulation of astroglial proliferation mediated by nucleotides. In highly enriched astroglial cultures, adenosine-5?-triphosphate (ATP), adenosine 5?-O-(3-thio)-triphosphate (ATP?S), adenosine 5?-O-(3-thio)-diphosphate (ADP?S; 0.01?1?mM), and adenosine-5?-diphosphate (ADP; 0.1?1?mM) increased proliferation up to 382%, an effect abolished in co-cultures containing 8% of microglia. The loss of ATP proliferative effect in co-cultures is supporte...

2011-01-01

Mitochondrial Potassium ATP Channels and Retinal Ischemic Preconditioning

Removal of lead from solution using non-living residual brewery yeast

PURPOSETo examine the mechanisms of ischemic preconditioning (IPC) related to the opening of mitochondrial KATP (mKATP) channels in the retina.Full Text Available

2006-05-01

Genetical approach to oxygen toxicity. [Saccharomyces cerevisiae

A number of preparations of residual non-living brewery yeast were examined for their ability to remove lead from solution. Those preparations included washed and un-washed intact yeast and washed and un-washed homogenates of the yeast cells. Using biosorption isotherm analysis it was found that the washed and un-washed preparations of intact, non-living yeast exhibited maximum biosorption capacities for lead of 127 and 99 mg/g dry weight biomass, respectively. The washed and un-washed cell homogenates exhibited maximum biosorption capacities of 38 and 139 mg lead/g dry weight biomass, respectively. Since it had previously been shown that these preparations of biomass were capable of removing uranium from solution by combined biosorption and precipitation processes, it was decided to examine removal of lead from solution using a form of equilibrium dialysis in which the biomass was retained within a ...

1998-10-01

Sirtuins, Bioageing, and Cancer

The role of intermediate products of dioxygen reduction in cytotoxic effects ascribed to oxygen molecules was studied in vivo using various yeast mutants with changed response to oxygen stress. It has been documented that superoxide radical exerts its deleterious effects on yeast cells directly and the role of other oxygen species derived from it is hardly detectable. Yeast Saccharomyces cerevisiae, however, cannot be considered as a typical eukaryotic organism due to its inability of synthesizing polyunsaturated fatty acids (pufa). These fatty acids are known as main target molecules during oxidative stress and their peroxidation leads to cytotoxic effects. As fatty acid content could be easily manipulated in yeast, this organism was used to evaluate the contribution of pufa peroxidation process to the cytotoxic effects of oxygen. Results obtained show, that yeast cells containing ...

1986-01-01

Iodination of glyceraldehyde 3-phosphate dehydrogenase

The Sirtuins are a family of orthologues of yeast Sir2 found in a wide range of organisms from bacteria to man. They display a high degree of conservation between species, in both sequence and function,...Full Text Available

Effect of Photosynthetic Bacterial Addition to Chlorella or ${\\omega}-Yeast$ on Growth of Rotifer, Brachionus plicatilis, and its Dietary Value for Flounder, Paralichthys olivaceus, Larvae

1. A high degree of homology in the positions of tyrosine residues in glyceraldehyde 3-phosphate dehydrogenase from lobster and pig muscle, and from yeast, prompted an examination of the reactivity...Full Text Available

1970-09-01

Sucrose accumulation in mature sweet melon fruits. [Cucumis melo

Full Text Available

2000-03-01

Pore properties and pharmacological features of the P2X receptor channel in airway ciliated cells

Mesocarp tissue from sucrose-accumulating sweet melon (Cucumis melo cv. Galia) showed sucrose synthase activity (ca 1 nkat/gfw) while soluble acid invertase and sucrose phosphate synthase activities were not observed. Sucrose uptake into mesocarp discs was linear with sucrose concentration (1-500 mM) and unaffected by PCMBS and CCCP. Sucrose compartmentation into the vacuole also increased linearly with sucrose concentration as indicated by compartmental efflux kinetics. Mesocarp discs incubated in /sup 14/C-fructose + UDP-glu synthesized /sup 14/C-sucrose and efflux kinetics indicated that the /sup 14/C-sucrose was compartmentalized. These data support the hypothesis that two mechanisms are involved in sucrose accumulation in sweet melon: (1) compartmentation of intact sucrose and (2) synthesis of sucrose via sucrose synthase and subsequent compartmentation in the vacuole.

1987-04-01

Interaction of Extracellular Domain 2 of the Human Retina-specific ATP-binding Cassette Transporter (ABCA4) with All-trans-retinal*

Airway ciliated cells express an ATP-gated P2X receptor channel of unknown subunit composition (P2X_cilia) which is modulated by Na⁺ and by long exposures to ATP. P2X_cilia...Full Text Available

2006-03-15

Evaluation of indexes for tumor growth and effect of radiation therapy using MRS; Experimental studies on SCCVII tumor bearing mice

The retina-specific ATP-binding cassette (ABC) transporter, ABCA4, is essential for transport of all-trans-retinal from the rod outer segment discs in the retina and is associated with...Full Text Available

2010-06-18

Differential use of two cyclic electron flows around photosystem I for driving CO₂-concentration mechanism in C₄ photosynthesis

Changes in the phosphate metabolism of the transplanted SCCVII (squamous cell carcinoma VII) tumor were studied before and after irradiation using in vivo [sup 31]P-magnetic resonance spectroscopy (MRS). Phosphocreatine (PCr), [gamma]-, [alpha]-, [beta]-ATP, inorganic phosphate (Pi), phosphomonoester (PME), and phosphodiester (PDE) were detected in [sup 31]P-MR spectra of tumor tissues. The curve fitting method was employed for calculating each peak of [sup 31]P-MR spectra (PCr, [beta]-ATP, Pi, PME, PDE). The values of PCr, PCr/Pi, [beta]-ATP/Pi and pH decreased in proportion to the growth of tumor, whereas that of Pi, PME and PDE increased. But the changes of [beta]-ATP were minimal in degree. Based on the fact that the ratio of PCr/Pi highly correlated with tumor volume and showed the most remarkable change among various parameters, it might be the useful index for the tumor growth. Because the ...

1992-03-01

Diagnostic value of adenosine-triphosphate radionuclide ventriculography in coronary artery disease. Comparison with conventional exercise stress method

Whereas linear electron flow (LEF) in photosynthesis produces both ATP and NADPH, the cyclic electron flow (CEF) around photosystem I has been shown to produce only ATP. Two alternative routes have...Full Text Available

2005-11-15

ATP Hydrolysis in Eg5 Kinesin Involves a Catalytic Two-water Mechanism*^?

The purpose of this study was to assess the effect of adenosine-triphosphate (ATP) on hemodynamics and left ventricular (LV) function, and to examine the value of ATP radionuclide-ventriculography (RNVG) for detecting coronary artery disease (CAD) in comparison with standard exercise RNVG. Thirty-eight consecutive patients with suspected CAD underwent ATP (0.18 mg/Kg/min) RNVG and symptom-limited exercise RNVG. Coronary angiography revealed 27 patients with CAD (Group S) and 11 without CAD (Group N). In group S, exercise induced an increase in LV end-diastolic volume (LVEDV) and LV end-systolic volume (LVESV), and a decrease in LV ejection fraction (LVEF). In contrast, ATP infusion caused LVESV to decrease in both groups, improving LVEF. Only 14 of 39 segments whose wall motion was deteriorated by exercise showed worsening regional wall motion during ATP infusion. The sensitivity ...

1995-07-01

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

Transient increase in glucose 1,6-bisphosphate in human skeletal muscle during isometric contraction.

Motor proteins couple steps in ATP binding and hydrolysis to conformational switching both in and remote from the active site. In our kinesin·AMPPPNP crystal structure, closure of the active...Full Text Available

2010-02-19

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

Changes in glucose 1,6-bisphosphate and regulators of glucose-1,6-bisphosphate synthase and phosphatase during isometric contraction have been determined. Biopsies were obtained from the quadriceps...Full Text Available

1989-03-15

Prostaglandin H₂ synthase-1 and -2 expression in guinea pig gestational tissues during late pregnancy and parturition

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

Polyketide synthase chemistry does not direct biosynthetic divergence between 9- and 10-membered enediynes

Increased intrauterine prostaglandin (PG) production is crucial for the initiation of parturition. To investigate the mechanisms controlling intrauterine PG synthesis, we examined the expression of...Full Text Available

2005-12-15

Mycophenolic acid inhibits activation of inducible nitric oxide synthase in rodent fibroblasts

Enediynes are potent antitumor antibiotics that are classified as 9- or 10-membered according to the size of the enediyne core structure. However, almost nothing is known about enediyne core biosynthesis,...Full Text Available

2010-06-22

Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq

Mycophenolate mofetil (MMF) is an immunosuppressive drug that acts as a selective inhibitor of inosine monophosphate dehydrogenase (IMPDH). MMF has recently been shown to inhibit the enzymatic activity...Full Text Available

2003-05-01

Identification of Phloem Involved in Assimilate Loading in Leaves by the Activity of the Galactinol Synthase Promoter1

BackgroundIsopentenyl diphosphate (IPP), a common biosynthetic precursor to the labdane diterpene forskolin, has been biosynthesised via a non-mevalonate pathway. Geranylgeranyl...Full Text Available

Genetic Association Analysis of NOS1 and Methamphetamine-Induced Psychosis Among Japanese

The definition of “minor” veins in leaves is arbitrary and of uncertain biological significance. Generally, the term refers to the smallest vein classes in the leaf, believed to function...Full Text Available

2000-07-01

Endogenous Nitric Oxide Protects Bacteria Against a Wide Spectrum of Antibiotics

The neuronal nitric oxide synthase gene (NOS1) is located at 12q24, a susceptibility region for schizophrenia, and produces nitric oxide (NO). NO has been reported to play important...Full Text Available

2011-03-01

Detection and Identification of Bartonella Species Pathogenic for Humans by PCR Amplification Targeting the Riboflavin Synthase Gene (ribC)

Bacterial nitric oxide synthases (bNOS) are present in many Gram-positive species and have been demonstrated to synthesize NO from arginine in vitro and in vivo. However, the physiological role...Full Text Available

2009-09-11

Characterization of Geraniol Synthase from the Peltate Glands of Sweet Basil1

Several Bartonella species have now been implicated as human pathogens. The recovery of these fastidious organisms in the clinical microbiology laboratory remains difficult, and current...Full Text Available

2003-03-01

A Bifunctional Geranyl and Geranylgeranyl Diphosphate Synthase Is Involved in Terpene Oleoresin Formation in Picea abies1[W][OA

The monoterpene fraction of the lemon-scented sweet basil (Ocimum basilicum) cv Sweet Dani consists mostly of citral (a mixture of geranial and neral), with lower levels of geraniol...Full Text Available

2004-01-01

Does adenine incorporation into nucleic acids measure total microbial production:: a response to comments by Fuhrman et al

The conifer Picea abies (Norway spruce) defends itself against herbivores and pathogens with a terpenoid-based oleoresin composed chiefly of monoterpenes (C₁₀) and diterpenes...Full Text Available

2010-02-01

The Functionally Conserved Nucleoporins Nup124p from Fission Yeast and the Human Nup153 Mediate Nuclear Import and Activity of the Tf1 Retrotransposon and HIV-1 VprV?

The uptake and incorporation of (/sup 3/H)adenine as a measure of total microbial (bacteria and unicellular algae) nucleic acid synthesis depends on the validity of several assumptions. A unique characteristic of the (/sup 3/H)adenine method is the ability to measure the specific radioactivity (nCi pmol/sup -1/) of the immediate precursor pool of the adenine incorporated into nucleic acids. This measurement permits correction for isotope dilution when the added radiotracer mixes with exogenous pools of structurally related compounds before uptake and with endogenous pools after transport into the cells. The immediate precursors to (/sup 3/H)adenine incorporation into cellular RNA and DNA are ATP and dATP. Since the intracellular dATP pool is in isotopic equilibrium with the ATP pool, one can determine the specific radioactivity of both precursor pools by measuring that of the intracellular ...

1986-11-01

Morphological Instabilities in a growing Yeast Colony Experiment and Theory

We report that the fission yeast nucleoporin Nup124p is required for the nuclear import of both, retrotransposon Tf1-Gag as well as the retroviral HIV-1 Vpr. Failure to import Tf1-Gag into the nucleus...Full Text Available

2005-04-01

CERN Document Server

We study the growth of colonies of the yeast Pichia membranaefaciens on agarose film. The growth conditions are controlled in a setup where nutrients are supplied through an agarose film suspended over a solution of nutrients. As the thickness of the agarose film is varied, the morphology of the front of the colony changes. The growth of the front is modeled by coupling it to a diffusive field of inhibitory metabolites. Qualitative agreement with experiments suggests that such a coupling is responsible for the observed instability of the front.

1997-01-01

Kes1p shares homology with human oxysterol binding protein and participates in a novel regulatory pathway for yeast Golgi-derived transport vesicle biogenesis.

Dissecting toxin immunity in virus-infected killer yeast uncovers an intrinsic strategy of self-protection

The yeast phosphatidylinositol transfer protein (Sec14p) is required for biogenesis of Golgi-derived transport vesicles and cell viability, and this essential Sec14p requirement is abrogated by inactivation...Full Text Available

1996-12-02

Toxin-secreting “killer” yeasts were initially identified >40 years ago in Saccharomyces cerevisiae strains infected with a double-stranded RNA “killer”...Full Text Available

2006-03-07

DNA repair genes

Arginine aminoacylation identity is context-dependent and ensured by alternate recognition sets in the anticodon loop of accepting tRNA transcripts.

Fission yeast S. pombe is assumed to be a good model for cloning of human DNA repair genes, because human gene is normally expressed in S. pombe and has a very similar protein sequence to yeast protein. We have tried to elucidate the DNA repair mechanisms of S. pombe as a model system for those of mammals. (J.P.N.)

1995-12-01

100

Amino acid sequences that determine the nuclear localization of yeast histone 2B.

Yeast arginyl-tRNA synthetase recognizes the non-modified wild-type transcripts derived from both yeast tRNA(Arg) and tRNA(Asp) with equal efficiency. It discriminates its cognate natural substrate,...Full Text Available

1996-09-16

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101

A heteromeric complex containing the centromere binding factor 1 and two basic leucine zipper factors, Met4 and Met28, mediates the transcription activation of yeast sulfur metabolism.

Histone-beta-galactosidase protein fusions were used to identify the domain of yeast histone 2B, which targets this protein to the nucleus. Amino acids 28 to 33 in H2B were required for nuclear localization...Full Text Available

1987-11-01

102

Depression of calcium pump activity in renal cortex of vitamin D-deficient rats with secondary hyperparathyroidism

Transcription activation of sulfur metabolism in yeast is dependent on two DNA binding factors, the centromere binding factor 1 (Cbf1) and Met4. While the role of Met4 was clearly established by showing...Full Text Available

1996-05-15

103

Depression of calcium pump activity in renal cortex of vitamin D-deficient rats with secondary hyperparathyroidism

To examine the hormonal regulation of the ATP-dependent Ca{sup 2+} pump in the kidneys, the ATP-dependent Ca{sup 2+} uptake by the basolateral membrane vesicles in the renal cortex was measured using radioactive calcium ({sup 45}Ca{sup 2+}) in rats with vitamin D deficiency or rats undergoing thyroparathyroidectomy. The V{sub max} of the Ca{sup 2+} pump activity was increased not only by administering calcitriol, but also by normalizing the serum calcium level in vitamin D-deficient rats. PTH suppressed the Ca{sup 2+} pump activity in normocalcemic vitamin D-deficient rats. Thyroparathyroidectomy did not affect the Ca{sup 2+} pump activity in the kidneys of normal rats. It was concluded that the ATP-dependent Ca{sup 2+} pump activity was depressed by secondary hyperparathyroidism in vitamin D-deficient rats. (author).

1990-01-01

104

Morphologie des cellules de levure et la reproduction sexuelle - Apercu general et quelques considerations

To examine the hormonal regulation of the ATP-dependent Ca"2"+ pump in the kidneys, the ATP-dependent Ca"2"+ uptake by the basolateral membrane vesicles in the renal cortex was measured using radioactive calcium ("4"5Ca"2"+) in rats with vitamin D deficiency or rats undergoing thyroparathyroidectomy. The V_m_a_x of the Ca"2"+ pump activity was increased not only by administering calcitriol, but also by normalizing the serum calcium level in vitamin D-deficient rats. PTH suppressed the Ca"2"+ pump activity in normocalcemic vitamin D-deficient rats. Thyroparathyroidectomy did not affect the Ca"2"+ pump activity in the kidneys of normal rats. It was concluded that the ATP-dependent Ca"2"+ pump activity was depressed by secondary hyperparathyroidism in vitamin D-deficient rats. (author).

105

British Library Electronic Table of Contents (United Kingdom)

Over the decades, basic research in life sciences has profited greatly from the study of the small unicellular fungal species Saccharomyces cerevisiae. This yeast turned out to be key for the identification and understanding of molecular mechanisms that underlay the basic functions of all eukaryotic cells. These include, but are not limited to, the regulatory mechanisms behind cellular reproduction (cell cycle control), cellular morphogenesis (cell polarity, cytoskeleton and membrane trafficking) and the management of cellular information (chromosome biology, transcription and translation). Rapid access to genomic information of many yeast species, combined with bioinformatics analyses, provide information on the evolutionary history of yeasts and the molecular ancestry of their constituen...

2011-01-01

106

Alcohol fermentation in olive oil extraction effluents

Triosephosphates Modulate Leaf Mitochondrial Phosphorylation by Inhibition and Uncoupling of Electron Transport ¹

Eight culture-collection yeast strains of various species and five newly isolated stains were tested for both growth in olive oil extraction effluents and fermentation of the sugars in the same media. The culture-collection yeast strains did not grow in an effluent containing 2.86% sugar (w/v), 8 g/litre phenolic substances, 4.58 g/litre titratable acidity and pH 4.96, whereas the newly isolated strains of Torulopsis sp. MK-1, Saccharomyces norbensis MC-1, S. oleaceus MC-2 and S. oleaginosus grew well and fermented the sugars. In the medium mentioned above, they produced alcohol in amounts of 1.63 to 1.38%, respectively. None of the yeasts grew in an olive oil extraction effluent vacuum-concentrated to over 13-14% of dry matter. The strain of T. sp. MK-1 showed a higher stability.

1989-01-01

107

Transport of Arginine and Aspartic Acid into Isolated Barley Mesophyll Vacuoles ¹

The effect of TP (triosephosphates:glyceraldehyde-3 phosphate, GAP, +dihydroxyacetone phosphate, DHAP) on respiration, phosphorylation and matrix ATP/ADP ratios of isolated oat mesophyll mitochondria...Full Text Available

1985-11-01

108

The efflux of a fluorescent probe is catalyzed by an ATP-driven extrusion system in Lactococcus lactis.

The transport of arginine into isolated barley (Hordeum vulgare L.) mesophyll vacuoles was investigated. In the absence of ATP, arginine uptake was saturable with a K_m...Full Text Available

1991-10-01

109

The accuracy and reverse effects of thallium myocardial SPECT using adenosine triphosphate loading in the diagnosis of coronary artery disease. Comparison with other loading methods semiquantitatively

Many bacteria, both gram positive and gram negative, extrude in an energy-dependent manner the fluorescent pH indicator 2',7'-bis-(2-carboxyethyl)-5[and -6]-carboxyfluorescein (BCECF) (D. Molenaar,...Full Text Available

1992-05-01

110

The DEXD/H-box RNA Helicase DDX19 Is Regulated by an ?-Helical Switch*S?

The adverse effects and diagnostic accuracy of thallium-201 myocardial SPECT after intravenous infusion of adenosine triphosphate (ATP) were studied and compared with SPECT examinations with other type of loading. The subjects of the study were 147 patients with or without coronary artery disease, who underwent some type of loading SPECT and coronary arteriography (CAG) within 30 days. Myocardial ischemia was evaluated qualitatively in SPECT and was compared with CAG for the diagnostic accuracy of coronary artery disease. The degree of myocardial uptake defect was also calculated semiquantitatively using visual scoring method and compared with coronary artery severity score. The adverse effects occurred in 46.7% of ATP loading SPECT which was more frequent that DIP loading SPECT, but all of them were transient and mild. As for the diagnostic ability the ATP loading SPECT was as effective as other type of loading in ...

1999-09-01

111

Rapid Phytochrome-mediated Changes in Adenosine 5?-Triphosphate Content of Etiolated Bean Buds ¹

DEXD/H-box RNA helicases couple ATP hydrolysis to RNA remodeling by an unknown mechanism. We used x-ray crystallography and biochemical analysis of the human DEXD/H-box...Full Text Available

2009-04-17

112

P2X purinoceptors mediate an endothelium-dependent hyperpolarization in longitudinal smooth muscle of anterior mesenteric artery in young chickens

This study was designed to determine the effects of red and far red irradiation on ATP metabolism in etiolated bean buds (Phaseolus vulgaris L. var. Red Kidney). Compared to dark controls,...Full Text Available

1974-01-01

113

Molecular characterization of cytoplasmic male sterility conditioned by Gossypium harknessii cytoplasm (CMS-D2) in upland cotton

Background and purpose:The chicken anterior mesenteric artery contains an outer longitudinal smooth muscle layer, whose neural regulation remains to be elucidated. ATP evokes a depolarization...Full Text Available

2009-10-01

114

British Library Electronic Table of Contents (United Kingdom)

Cytoplasmic male sterility (CMS) is a maternally inherited trait that fails to produce functional pollen grains. The CMS system is widely employed to facilitate the utilization of heterosis in major crops. However, little is known about the CMS associated genes in Upland cotton (Gossypium hirsutum). The objective of this study was to compare CMS cotton (CMS-D2) with the cytoplasm from G. harknessii and its isogenic maintainer line with the normal fertile Upland cotton cytoplasm to identify CMS-D2 specific gene(s) and to develop CMS-specific sequence characterized amplified region (SCAR) markers. Based on Southern blot analysis using 10 mitochondrial gene-specific probes (cob, cox2, atp6, atp9, nad3, cox3, atpA, cox1, nad6 and nad9), three probes (cox3, atpA, and nad6) revealed restriction ...

2011-01-01

115

Kinetics of nucleotide transport in rat heart mitochondria studied by a rapid filtration technique

Increased expression of cholesterol transporter ABCA1 is highly correlated with severity of dementia in AD hippocampus

A rapid filtration technique has been used to measure at room temperature the kinetics of ADP and ATP transport in rat heart mitochondria in the millisecond time range. Transport was stopped by cessation of the nucleotide supply, without the use of a transport inhibitor, thus avoiding any quenching delay. The kinetics of ({sup 14}C)ADP transport in energized mitochondria were apparently monophasic. The rate of transport of ({sup 14}C)ATP in energized mitochondria was 5-10 times lower than that of ({sup 14}C)ADP. Upon uncoupling, the rate of ({sup 14}C)ATP uptake was enhanced, and that of ({sup 14}C)ADP uptake was decreased. However, the two rates did not equalize, indicating that transport was not exclusively electrogenic. Transport of ({sup 14}C)ADP and ({sup 14}C)ATP by resting mitochondria followed biphasic kinetics. Depletion of nucleotides in resting mitochondria resulted in a greater decrease in ...

1990-10-01

116

Identification of genetic variation and haplotype structure of the canine ABCA4 gene for retinal disease association studies

To gain insight into ATP-binding cassette transporter A1 (ABCA1) function and its potential role in AD pathology, we analyzed the expression of the cholesterol transporter ABCA1...Full Text Available

2010-03-08

117

Gyrase B Inhibitor Impairs HIV-1 Replication by Targeting Hsp90 and the Capsid Protein*

Over 200 mutations in the retina specific member of the ATP-binding cassette transporter super-family (ABCA4) have been associated with a diverse group of human retinal diseases....Full Text Available

2010-10-01

118

Energy metabolism of the untrained muscle of elite runners as observed by 31P magnetic resonance spectroscopy: evidence suggesting a genetic endowment for endurance exercise.

Chemical genetics is an emerging approach to investigate the biology of host-pathogen interactions. We screened several inhibitors of ATP-dependent DNA motors and detected the gyrase B inhibitor coumermycin...Full Text Available

2010-12-10

119

Efficient Phagocytosis Requires Triacylglycerol Hydrolysis by Adipose Triglyceride Lipase*

The purpose of this study was to investigate whether genetically determined properties of muscle metabolism contribute to the exceptional physical endurance of world-class distance runners. ATP, phosphocreatine,...Full Text Available

1988-12-01

120

Diterpene Biosynthesis in Maize Seedlings in Response to Fungal Infection ¹

Macrophage phagocytosis is an essential biological process in host defense and requires large amounts of energy. To date, glucose is believed to represent the prime substrate for ATP production in macrophages....Full Text Available

2010-06-25

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121

Cytosolic Phosphofructokinase from Spinach Leaves ¹

A cell-free system which catalyzes the biosynthesis of terpene hydrocarbons when supplemented with mevalonate, Mn²⁺, and ATP was prepared from the scutellum-embryonic axis region of maize...Full Text Available

1979-09-01

122

Conditional deletion of Abca3 in alveolar type II cells alters surfactant homeostasis in newborn and adult mice

Cytosolic ATP-phosphofructokinase (PFK) from spinach leaves (Spinacia oleracea L.) was inhibited by submillimolar concentrations of free Mg²⁺. The free Mg²⁺ concentration...Full Text Available

1989-08-01

123

Comparison of McAuley/fasting insulin indices with ATP III clinical criteria for the diagnosis of insulin resistance in type 2 diabetes mellitus

ATP-binding cassette A3 (ABCA3) is a lipid transport protein required for synthesis and storage of pulmonary surfactant in type II cells in the alveoli. Abca3 was conditionally deleted...Full Text Available

2010-05-01

124

Chemiosmotic model for plant mitochondria

Objective:To estimate the prevalence of insulin resistant syndrome (IRS) among newly diagnosed patients with type 2 diabetes and to test their validity against two indices...Full Text Available

2011-07-01

125

Anaerobic Growth Yields of Aerobacter cloacae and Escherichia coli

A balanced application of chemiosmosis to plant mitochondria is presented. Electron transport carriers are sequenced to achieve proton translocation across the inner membrane. The processes of proton translocation, use, and release are integrated to yield a description of the effective proton gradient available for metabolite transport and ATP synthesis. 27 references, 6 figures.

1983-12-01

126

Adenosine 5?-triphosphate sulphurylase from Saccharomyces cerevisiae

Aerobacter cloacae UW-C83 and Escherichia coli K-12 were grown under various anaerobic environments. Y_atp values were calculated by determination...Full Text Available

1967-10-01

127

A TRAP Transporter for Pyruvate and Other Monocarboxylate 2-Oxoacids in the Cyanobacterium Anabaena sp. Strain PCC 7120?

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

128

Synthesis of high specific activity tritium labelled [2-{sup 3}H]-adenosine-5`-triphosphate

In the cyanobacterium Anabaena sp. strain PCC 7120, open reading frames (ORFs) alr3026, alr3027, and all3028 encode a tripartite ATP-independent...Full Text Available

2010-11-01

129

Mapping of the nucleotide-binding sites in the ADP/ATP carrier of beef heart mitochondria by photolabeling with 2-azido[#alpha#-"3"2P]adenosine diphosphate

A procedure for high level tritium labelling at the C2-H position of adenosine 5`-triphosphate ([2-{sup 3}H]-ATP, 1), based on the tritiodehalogenation reaction of 2-bromoadenosine 5`-triphosphate (2) has been elaborated. This precursor was prepared in a six-step synthesis from guanosine. The tritiodehalogenation of (2) for three hours over palladium oxide in phosphate buffer yielded tritium labelled ATP with high specific activity, in good chemical yield. (author).

1996-08-01

130

Bacterial radiosensitization by using radiation processing in combination with essential oil: Mechanism of action

2-Azido[#alpha#-"3"2P]adenosine diphosphate (2-azido[#alpha#-"3"2P]ADP) has been used to photolabel the ADP/ATP carrier in beef heart mitochondria. In reversible binding assays carried out in the dark, this photoprobe was found to inhibit ADP/ATP transport in beef heart mitochondria and to bind to two types of specific sites of the ADP/ATP carrier characterized by high-affinity binding (K/sub d/ = 20 #mu#M) and low-affinity binding (K/sub d/ = 400 #mu#M). In contrast, it was unable to bind to specific carrier sites in inverted submitochondrial particles. Upon photoirradiation of beef heart mitochondria in the presence of 2-azido[#alpha#-"3"2P]ADP, the ADP/ATP carrier was covalently labeled. After purification, the photolabeled carrier protein was cleaved chemically by acidolysis or cyanogen bromide and enzymatically with the Staphylococcus aureus V8 protease. In the ADP/ATP carrier ...

131

Variants within the yeast Ty sequence family encode a class of structurally conserved proteins.

Spice extracts under the form of essential oils were tested for their efficiency to increase the relative radiosensitivity of Listeria monocytogenes and Escherichia coli O157H7 in culture media. The two pathogens were treated by gamma-irradiation alone or in combination with oregano essential oil to evaluate their mechanism of action. The membrane murein composition, and the intracellular and extracellular concentration of ATP was determined. The bacterial strains were treated with two irradiation doses: 1.2 kGy to induce cell damage and 3.5 kGy to cause cell death for L. monocytogenes. A dose of 0.4 kGy to induce cell damages, 1.1 kGy to obtain viable but nonculturable (VBNC) state and 1.3 kGy to obtain a lethal dose was also applied on E. coli O157H7. Oregano essential oil was used at 0.020% and 0.025% (w/v), which is the minimum inhibitory concentration (MIC) for L. monocytogenes. For E. coli O157H7, a concentration of 0.006% and 0.025% (w/v) which is the ...

2009-07-15

132

The requirement of yeast replication origins for pre-replication complex proteins is modulated by transcription

The Ty transposable elements of Saccharomyces cerevisiae form a heterogeneous family within which two broad structural classes (I and II) exist. The two classes differ by two large substitutions and...Full Text Available

1985-06-11

133

The recruitment of acetylated and unacetylated tropomyosin to distinct actin polymers permits the discrete regulation of specific myosins in fission yeast

The mini-chromosome maintenance proteins Mcm2–7 are essential for DNA replication. They are loaded onto replication origins during G1 phase of the cell cycle to form a pre-replication complex...Full Text Available

2005-01-01

134

The fission yeast gene pmt1+ encodes a DNA methyltransferase homologue.

Tropomyosin (Tm) is a conserved dimeric coiled-coil protein, which forms polymers that curl around actin filaments in order to regulate actomyosin function. Acetylation of the Tm N-terminal...Full Text Available

2010-10-01

135

Structure-Function Study of the N-terminal Domain of Exocyst Subunit Sec3*

DNA methylation of cytosine residues is a widespread phenomenon and has been implicated in a number of biological processes in both prokaryotes and eukaryotes. This methylation occurs at the 5-position...Full Text Available

1995-01-25

136

Some highlights of research on aging with invertebrates, 2010

The exocyst is an evolutionarily conserved octameric complex involved in polarized exocytosis from yeast to humans. The Sec3 subunit of the exocyst acts as a spatial landmark for exocytosis through...Full Text Available

2010-04-02

137

SIRT1 contributes to telomere maintenance and augments global homologous recombination

This annual review focuses on invertebrate model organisms, which continue to yield fundamental new insights into mechanisms of aging. This year, the budding yeast has been used to understand how asymmetrical...Full Text Available

2011-02-01

138

PtdIns 3-Kinase Orchestrates Autophagosome Formation in Yeast

Yeast Sir2 deacetylase is a component of the silent information regulator (SIR) complex encompassing Sir2/Sir3/Sir4. Sir2 is recruited to telomeres through Rap1, and this complex spreads into subtelomeric...Full Text Available

2010-12-27

139

Overexpression of human virus surface glycoprotein precursors induces cytosolic unfolded protein response in Saccharomyces cerevisiae

Eukaryotic cells can massively transport their own cytoplasmic contents into a lytic compartment, the vacuole/lysosome, for recycling through a conserved system called autophagy. The key process in...Full Text Available

2011-01-01

140

O2-dependent methionine auxotrophy in Cu,Zn superoxide dismutase-deficient mutants of Saccharomyces cerevisiae.

BackgroundThe expression of human virus surface proteins, as well as other mammalian glycoproteins, is much more efficient in cells of higher eukaryotes rather than yeasts. The limitations...Full Text Available

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141

Nuclear Pore Complex Number and Distribution throughout the Saccharomyces cerevisiae Cell Cycle by Three-Dimensional Reconstruction from Electron Micrographs of Nuclear Envelopes

Mutant strains of the yeast Saccharomyces cerevisiae which lack functional Cu,Zn superoxide dismutase (SOD-1) do not grow aerobically unless supplemented with methionine. The molecular basis of this...Full Text Available

1990-04-01

142

Mitochondrial transmission during mating in Saccharomyces cerevisiae is determined by mitochondrial fusion and fission and the intramitochondrial segregation of mitochondrial DNA.

The number of nuclear pore complexes (NPCs) in individual nuclei of the yeast Saccharomyces cerevisiae was determined by computer-aided reconstruction of entire nuclei from electron...Full Text Available

1997-11-01

143

Mammalian Sirtuins and Energy Metabolism

To gain insight into the process of mitochondrial transmission in yeast, we directly labeled mitochondrial proteins and mitochondrial DNA (mtDNA) and observed their fate after the fusion of two cells....Full Text Available

1997-07-01

144

MAPPING THE INITIATOR BINDING TAF2 SUBUNIT IN THE STRUCTURE OF HYDRATED YEAST TFIID

Sirtuins are highly conserved NAD⁺-dependent protein deacetylases and/or ADP-ribosyltransferases that can extend the lifespan of several lower model organisms including yeast, worms and flies....Full Text Available

145

Legionella fairfieldensis sp. nov. isolated from cooling tower waters in Australia.

SummaryThe general transcription factor TFIID is a large multi-subunit complex required for the transcription of most protein-encoding genes by RNA polymerase II. Taking advantage...Full Text Available

2009-03-11

146

Insights into Cdc13 dependent telomere length regulation

Three Legionella-like organisms were isolated from water from the cooling towers of two Australian institutions. The strains grew on buffered charcoal-yeast extract (BCYE) agar but not on BCYE agar...Full Text Available

1991-03-01

147

Increased Ethanol Productivity in Xylose-Utilizing Saccharomyces cerevisiae via a Randomly Mutagenized Xylose Reductase?

Cdc13 is a single stranded telomere binding protein that specifically localizes to the telomere ends of budding yeasts and is essential for cell viability. It caps the ends of chromosomes thus preventing...Full Text Available

148

Improving yield of industrial biomass propagation by increasing the Trx2p dosage

Baker's yeast (Saccharomyces cerevisiae) has been genetically engineered to ferment the pentose sugar xylose present in lignocellulose biomass. One of the reactions controlling the...Full Text Available

2010-12-01

149

Identification of Potential Calorie Restriction-Mimicking Yeast Mutants with Increased Mitochondrial Respiratory Chain and Nitric Oxide Levels

The beneficial effect of improving yeast redox response by increasing thioredoxin levels has been shown. Decreased lipid and protein oxidation is reflected in an increased biomass yield. In addition,...Full Text Available

2010-09-01

150

Fuzzy-decision-making problems of fuel ethanol production using a genetically engineered yeast

Calorie restriction (CR) induces a metabolic shift towards mitochondrial respiration; however, molecular mechanisms underlying CR remain unclear. Recent studies suggest that CR-induced mitochondrial...Full Text Available

151

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

A fuzzy-decision-making procedure is applied to find the optimal feed policy of a fed-batch fermentation process for fuel ethanol production using a genetically engineered Saccharomyces yeast 1400 (pLNH33). The policy consisted of feed flow rate, feed concentration, and fermentation time. The recombinant yeast 1400 (pLNH33) can utilize glucose and xylose simultaneously to produce ethanol. However, the parent yeast utilizes glucose only. A partially selective model is used to describe the kinetic behavior of the process. In this study, this partially selective fermentation process is formulated as a general multiple-objective optimal control problem. By using an assigned membership function for each of the objectives, the general multiple-objective optimization problem can be converted into a maximizing decision problem. In order to obtain a global solution, a hybrid method of differential evolution is introduced to solve ...

1998-08-01

152

Expansions, contractions, and fragility of the spinocerebellar ataxia type 10 pentanucleotide repeat in yeast

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

153

Exit from the Golgi Is Required for the Expansion of the Autophagosomal Phagophore in Yeast Saccharomyces cerevisiae

Spinocerebellar ataxia 10 (SCA10) is an autosomal dominant disease caused by large-scale expansions of the (ATTCT)_n repeat within an intron of the human ATXN10 gene. In contrast...Full Text Available

2011-02-15

154

Engineering ligand-responsive RNA controllers in yeast through the assembly of RNase III tuning modules

The delivery of proteins and organelles to the vacuole by autophagy involves membrane rearrangements that result in the formation of large vesicles called autophagosomes. The mechanism underlying autophagosome...Full Text Available

2010-07-01

155

Cloning and Analysis of a Candida albicans Gene That Affects Cell Surface Hydrophobicity

The programming of cellular networks to achieve new biological functions depends on the development of genetic tools that link the presence of a molecular signal to gene-regulatory activity. Recently,...Full Text Available

2011-07-01

156

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

The opportunistic pathogenic yeast Candida albicans exhibits growth phase-dependent changes in cell surface hydrophobicity, which has been correlated with adhesion to host tissues....Full Text Available

2001-06-01

157

Biotransformation of Explosives by the Old Yellow Enzyme Family of Flavoproteins

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

158

Arabidopsis thaliana Chromosome 4 Replicates in Two Phases That Correlate with Chromatin State

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

159

Altering the ribosomal subunit ratio in yeast maximizes recombinant protein yield

DNA replication programs have been studied extensively in yeast and animal systems, where they have been shown to correlate with gene expression and certain epigenetic modifications. Despite the conservation...Full Text Available

2010-06-01

160

Affinity maturation of human botulinum neurotoxin antibodies by light chain shuffling via yeast mating

BackgroundThe production of high yields of recombinant proteins is an enduring bottleneck in the post-genomic sciences that has yet to be addressed in a truly rational manner. Typically...Full Text Available

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161

A Novel Form of Transcriptional Silencing by Sum1-1 Requires Hst1 and the Origin Recognition Complex

Botulism is caused by the botulinum neurotoxins (BoNTs), the most poisonous substance known. Because of the high potency of BoNT, development of diagnostic and therapeutic antibodies for botulism requires...Full Text Available

2010-04-01

162

Yeast as a Model System to Study Tau Biology

In the yeast Saccharomyces cerevisiae, a and α mating-type information is stored in transcriptionally silenced cassettes called HML and HMR....Full Text Available

2001-05-01

163

Cell cultures are more sensitive than Saccharamoyces cervisiae tests for assessing the toxicity of aquatic pollutants

Hyperphosphorylated and aggregated human protein tau constitutes a hallmark of a multitude of neurodegenerative diseases called tauopathies, exemplified by Alzheimer's disease. In spite of an enormous amount of research performed on tau biology, several crucial questions concerning the mechanisms of tau toxicity remain unanswered. In this paper we will highlight some of the processes involved in tau biology and pathology, focusing on tau phosphorylation and the interplay with oxidative stress. In addition, we will introduce the development of a human tau-expressing yeast model, and discuss some crucial results obtained in this model, highlighting its potential in the elucidation of cellular processes leading to tau toxicity.

2011-04-06

164

The effect of substrate modification on microbial growth on surfaces

Cultured fish and human cells have been used as bioassay systems for the evaluation of the toxicity of aquatic pollutants. Numerous assays using bacteria and yeast have also been used for such purposes. The authors report the toxicity of aquatic pollutants (Cd, Hg, and Ni), using cell culture systems and the yeast Saccharomyces cervisiae test. Cd, Hg, and Ni were chosen as model compounds of pollutants because the related toxicity is now fairly well established.

1988-07-01

165

Identification of actively filling sucrose sinks. [Solanum tuberosum; Phaseolus lunatus; Manihot esculenta; Liquidambar styraciflua L. ; Carya illinoinensis

The principle aim of the program was to produce a novel, non-leaching antimicrobial surface for commercial development and future use in the liquid food packaging industry. Antimicrobial surfaces which exist presently have been produced to combat the growth of prokaryotic organisms and usually function as slow release systems. A system which could inhibit eukaryotic growth without contaminating the surrounding 'environment' with the inhibitor was considered of great commercial importance. The remit of this study was concerned with creating a surface which could control the growth of eukaryotic organisms found in fruit juice with particular interest in the yeast, Saccharomyces cerevisiae. Putative antimicrobial surfaces were created by the chemical modification of the test substrate polymers; nylon and ethylvinyl alcohol (EVOH). Surfaces were chemically modified by the covalent coupling of antimicrobial agents known to be active against the ...

1998-07-01

166

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

Certain actively filling plant sucrose sinks such as a seed, a tuber, or a root can be identified by measuring the uridine diphosphate and pyrophosphate-dependent metabolism of sucrose. Sucrolysis in both active and quiescent sucrose sinks was tested and sucrose synthase was found to be the predominant sucrose breakdown activity. Sucrolysis via invertases was low and secondary in both types of sinks. Sucrose synthase activity dropped markedly, greater than fivefold, in quiescent sinks. The test are consistent with the hypothesis that the sucrose filling activity, i.e. the sink strength, of these plant sinks can be measured by testing the uridine diphosphate and pyrophosphate-dependent breakdown of sucrose. Measuring the initial reactions of sucrolysis shows much promise for use in agriculture crop and tree improvement research as a biochemical test for sink strength.

1989-04-01

167

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

168

Characterization of the mannan synthase promoter from guar (Cyamopsis tetragonoloba)

British Library Electronic Table of Contents (United Kingdom)

Guar seed gum, consisting primarily of a high molecular weight galactomannan, is the most cost effective natural thickener, having broad applications in the food, cosmetics, paper, pharmaceutical and petroleum industries. The properties of the polymer can potentially be enhanced by genetic modification. Development of suitable endosperm-specific promoters for use in guar is desirable for metabolic engineering of the seed gum. A ~1.6?kb guar mannan synthase (MS) promoter region has been isolated. The MS promoter sequence was fused with the GUS reporter gene and overexpressed in the heterologous species alfalfa (Medicago sativa). The potential strength and specificity of the MS promoter was compared with those of the constitutive 35S promoter and the seed specific ?-phaseolin promoter. Quant...

2011-01-01

169

c-JUN N-TERMINAL KINASE MODULATES OXIDANT STRESS AND PEROXYNITRITE FORMATION INDEPENDENT OF INDUCIBLE NITRIC OXIDE SYNTHASE IN ACETAMINOPHEN HEPATOTOXICITY

Up-Regulation of the RhoA/Rho-Kinase Signaling Pathway in Corpus Cavernosum from Endothelial Nitric-Oxide Synthase (NOS), but Not Neuronal NOS, Null Mice

Acetaminophen (APAP) overdose, which causes liver injury in animals and humans, activates c-jun N-terminal kinase (JNK). Although it was shown that the JNK inhibitor SP600125 effectively reduced...Full Text Available

2010-07-01

170

Nucleotide sequence and genetic analysis of a 13.1-kilobase-pair Pseudomonas denitrificans DNA fragment containing five cob genes and identification of structural genes encoding Cob(I)alamin adenosyltransferase, cobyric acid synthase, and bifunctional cobinamide kinase-cobinamide phosphate guanylyltransferase.

We tested the hypothesis that the basal release of nitric oxide (NO) from endothelial cells modulates contractile activity in the corpus cavernosum (CC) via inhibition of the RhoA/Rho-kinase signaling...Full Text Available

2010-04-01

171

Insect-Induced Conifer Defense. White Pine Weevil and Methyl Jasmonate Induce Traumatic Resinosis, de Novo Formed Volatile Emissions, and Accumulation of Terpenoid Synthase and Putative Octadecanoid Pathway Transcripts in Sitka Spruce1[w

A 13.1-kb DNA fragment carrying Pseudomonas denitrificans cob genes has been sequenced. The nucleotide sequence and genetic analysis revealed that this fragment contained five different cob genes named...Full Text Available

1991-10-01

172

Expression Levels of Thymidylate Synthase, Thymidylate Phosphorylase and Dihydropyrimidine Dehydrogenase in Head and Neck Squamous Cell Carcinoma: Preliminary Study

Stem-boring insects and methyl jasmonate (MeJA) are thought to induce similar complex chemical and anatomical defenses in conifers. To compare insect- and MeJA-induced terpenoid responses, we analyzed...Full Text Available

2005-01-01

173

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.

IntroductionPharyngo-laryngeal tumors classified as T3-4, N0-3, M0, are conventionally treated by mutilating surgery (total (pharyngo)-laryngectomy). Neo-adjuvant chemotherapy with...Full Text Available

174

Both the Fas Ligand and Inducible Nitric Oxide Synthase Are Needed for Control of Parasite Replication within Lesions in Mice Infected with Leishmania major whereas the Contribution of Tumor Necrosis Factor Is Minimal

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

175

Serine-324 of myosin's heavy chain is photoaffinity-labeled by 3 prime (2 prime )-O-(4-benzoylbenzoyl)adenosine triphosphate

Following infection with the protozoan parasite Leishmania major, C57BL/6 mice develop a small lesion that heals spontaneously. Resistance to infection is associated with the development...Full Text Available

2003-09-01

176

Helminthosporium maydis T toxin decreased calcium transport into mitochondria of susceptible corn

A portion of the active site of rabbit skeletal myosin near the ribose ring of ATP can be labeled by the photoaffinity analogue 3{prime}(2{prime})-O-(4-benzoylbenzoyl)adenosine triphosphate (Bz{sub 2}ATP). The specificity of the photolabeling was assured by first trapping ({sup 14}C)Bz{sub 2}ATP at the active site by use of thiol cross-linking agents. Five radioactive peptides were isolated by high-performance liquid chromatography after extensive trypsin and subtilisin digestion of photolabeled myosin subfragment 1. Four of these peptides were sequenced by Edman techniques, and all originated from a region with the sequence Gly-Glu-Ile-Thr-Val-Pro-Ser-Ile-Asp-Asp-Gln, which corresponds to rabbit myosin heavy chain residues 312-328. The fifth labeled peptide had an amino acid composition appropriate for residues 312-328. Amino acid composition, radiochemical analysis, and sequence data indicate that Ser-324 is the major ...

1989-05-02

177

Motor Proteins Have Highly Correlated Brownian Engines

The effects of purified Helminthosporium maydis T (HmT) toxin on active Ca/sup 2 +/ transport into isolated mitochondria and microsomal vesicles were compared for a susceptible (T) and a resistant (N) strain of corn (Zea mays). ATP, malate, NADH, or succinate could drive /sup 45/Ca/sup 2 +/ transport into mitochondria of corn roots. Ca/sup 2 +/ uptake was dependent on the proton electrochemical gradient generated by the redox substrates or the reversible ATP synthetase, as oligomycin inhibited ATP-driven CA/sup 2 +/ uptake while KCN inhibited transport driven by the redox substrates. Purified native HmT toxin completely inhibited Ca/sup 2 +/ transport into T mitochondria at 5 to 10 nanograms per milliliter while transport into N mitochondria was decreased slightly by 100 nanograms per milliliter toxin. Malate-driven Ca/sup 2 +/ transport in T mitochondria was frequently more inhibited by 5 nanograms per milliliter toxin ...

1984-04-01

178

CERN Document Server

Two headed motor proteins, such as kinesin and dynein, hidrolyze environmental ATP in order to propel unidirectionally along cytoskeletal filaments such as microtubules. In the case of kinesin, protein heads bind primarily on the alpha tubulin site of asymmetric alpha-beta 8nm-long tubulin dimers that constitute the microtubular protofilaments. Kinesin dimers overcome local binding forces up to 5pN and are known to move on protofilaments with ATP concentration-dependent speeds while hydrolizing on average one ATP molecule per 8nm step. The salient features of protein trajectories are the distinct abrupt usually 8nm-long steps from one tubulin dimer to the next interlaced with long quiescent binding periods at a tubulin site. Discrete walks of this type are characterized by substantially reduced variances compared to pure biased random walks, and as a result rule out flashing-type ratchet models as possible mechanisms for ...

1998-01-01

179

Hydrolysis of adenosine 5'-triphosphate: an isotope-labeling study

Retrospective Monte Carlo dose calculations with limited beam weight information

A combination of /sup 18/O-labeling experiments and kinetic studies to clarify the nonenzymatic hydrolytic pathways of adenosine 5'-triphosphate (ATP) at pH values ranging from 0 to 8.3 has been used in this experiment. In 1 N and 0.1 N HCl, the data are consistent with the hypothesis that hydrolysis occurs by addition-elimination, with initial attack 93% ..gamma.. and 7% ..beta..; both lead only to ADP + P/sub i/. In the subsequent hydrolysis of the ADP to AMP + P/sub i/, attack is 83% ..beta.. and 17% ..cap alpha... At pH 8.3, the data are consistent with the hypothesis that hydrolsysis occurs by elimination-addition. Over the entire pH range studied, no oxygen exchange between water and ATP, ADP, or P/sub i/ was detected. Nonenzymatic hydrolysis and isotopic analysis of the resultant P/sub i/ comprise a preferred means of assaying the isotopic enrichment of (..gamma..-/sup 18/O)ATP to be used in studies of ...

1982-12-15

180

Yeast artificial chromosome libraries containing large inserts from mouse and human DNA

An important unresolved issue in outcomes analysis for lung complications is the effect of poor or completely lacking heterogeneity corrections in previously archived treatment plans. To estimate this effect, we developed a novel method based on Monte Carlo (MC) dose calculations which can be applied retrospectively to RTOG/AAPM-style archived treatment plans (ATP). We applied this method to 218 archived nonsmall cell lung cancer lung treatment plans that were originally calculated either without heterogeneity corrections or with primitive corrections. To retrospectively specify beam weights and wedges, beams were broken into Monte Carlo-generated beamlets, simulated using the VMC++ code, and mathematical optimization was used to match the archived water-based dose distributions. The derived beam weights (and any wedge effects) were then applied to Monte Carlo beamlets regenerated based on the patient computed tomography densities. Validation of the process was ...

2007-01-01

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181

Assessment of the role of oxygen and mitochondria in heat shock induction of radiation and thermal resistance in Saccharomyces cerevisiae

Yeast artificial chromosome (YAC) libraries have been difficult to construct with average insert sizes >400 kilobase pairs when DNA is size-fractionated in low-melting-point agarose. By using yeast chromosomes in mock cloning experiments, the authors found that polyamines should be present whenever agarose containing high molecular weight DNA is melted to protect DNA from degradation. By incorporating polyamines during the cloning procedure, they constructed YAC libraries from mouse and human DNA with average insert sizes of 700 and 620 kilobase pairs, respectively. Several genome equivalents of these YAC libraries were replicated onto the surface of many duplicate agar plates using a 40,000 multipin transfer device. High-density filter replicas were screened by hybridization, and 70 mouse YAC clones from 31 loci and 132 human YAC clones from 49 loci were isolated.

1991-05-15

182

Removal of uranium from solution using residual brewery yeast: combined biosorption and precipitation

In response to a heat shock, the yeast Saccharomyces cerevisiae undergoes a large increase in its resistance to heat and, by the induction of its recombinational DNA repair capacity, a corresponding increase in resistance to radiation. Yeast which lack mitochondrial DNA, mitochondria-controlled protein synthetic apparatus, aerobic respiration, and electron transport (rho/sup 0/ strain) were used to assess the role of O/sub 2/, mitochondria, and oxidative processes controlled by mitochondria in the induction of these resistances. We have found that rho/sup 0/ yeast grown and heat shocked in either the presence or absence of O/sub 2/ are capable of developing both radiation and heat resistance. We conclude that neither the stress signal nor its cellular consequences of induced heat and radiation resistance are directly dependent on O/sub 2/, mitochondrial DNA, or mitochondria-controlled protein synthetic or oxidative ...

1983-10-01

183

Pectinolytic yeast isolates for cold-active polygalacturonase production

Whilst unwashed preparations of biomass from a local brewery had an apparent maximum biosorption capacity for uranium of 360 mg/g (dry weight biomass) washing reduced this maximum to 150 mg/g. Homogenization of both biomass preparations and recovery of cellular debris had no significant effect on the maximum biosorption capacities although at lower equilibrium concentrations of uranium differences in the biosorption capacities were detected. When unwashed biomass was retained by a semi-permeable membrane 40% of uranium used in the experiments precipitated outside that membrane. Therefore a significant proportion of the uranium removed from solution, and previously attributed to biosorption by the yeast biomass, resulted from precipitation brought about by interaction with low molecular weight components loosely associated with the biomass. (Author).

1997-04-01

184

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

185

Microbiological quality and biophenol content of hot air-dried Thassos cv. table olives upon storage

British Library Electronic Table of Contents (United Kingdom)

Abstract Microbiological quality and biophenol content evolution was studied in minimally processed Thassos cv table olives by hot air dehydration under mild conditions (40C, 24-h, aw-=-0.893) and storage under characteristic packaging conditions (vacuum, 100% N2 and air) at 4 and 20C over a period of 180 days. No salt was used in the production line or packaging. The undesirable microorganisms (Enterobacteriaceae, Staphylococcus aureus, Bacillus, Clostridium) were undetectable until the end of the storage period. Also, modified atmospheres prevented fungal growth at both temperatures apart from the samples stored in air, in which Penicillium and Aspergillus spp. were identified. At 20C, a coexistence of mesophilic bacteria and yeasts occurred. At 4C, yeasts were the predominant microflora...

2011-01-01

186

Extrinsic allergic alveolitis induced by the yeast Debaryomyces hansenii.

Effects of dietary glucosylceramide on dermatitis in atopic dermatitis model mice

A 65-yr-old female developed cough, fever and dyspnoea following repeated exposure to a home ultrasonic humidifier. High-resolution computed tomography showed ground-glass opacity in both lung fields. Arterial blood gas analysis gave an oxygen tension of 8.38 kPa (63 Torr). Pulmonary function testing revealed restrictive ventilatory impairment with a reduction in the diffusing capacity. The diagnosis of extrinsic allergic alveolitis (EAA) was confirmed by radiographic findings, pathological evidence of alveolitis and reproductive development by a provocation test to the humidifier water. The yeast Debaryomyces Hansenii was the only microorganism cultured from the water of the humidifier. The double diffusion precipitating test and lymphocyte proliferative response was positive for an extract of D. Hansenii, providing evidence to incriminate this fungus. This is the first described case of EAA caused by D. Hansenii. PMID:12449192

2002-11-01

187

British Library Electronic Table of Contents (United Kingdom)

The effects of dietary plant and yeast cerebroside (glucosylceramide), a major sphingolipid in plants and yeast, on atopic dermatitis (AD) like symptoms were investigated in a mouse model. After 7 wk of feeding with a diet containing maize glucosylceramide, plasma IgE levels became significantly lower and in contrast, the levels of interleukin (IL)-12, which induces cellular immunity, became significantly higher in the AD mice than in the controls. However, the sphingolipid constituents of the skin fraction in the maize glucosylceramide fed group did not contain sphingoid bases of plant origin, such as 8-unsaturated sphingoid bases. The results of the present study indicated that dietary plant glucosylceramide prevented AD-like symptoms in AD model mice via regulation of Th1/Th2 balance. P...

2010-01-01

188

Structural features underlying selective inhibition of protein kinase CK2 by ATP site-directed tetrabromo-2-benzotriazole

Previous studies using in vitro cell culture systems have shown the role of the dynamin-related GTPase Opa1 in apoptosis prevention and mitochondrial DNA (mtDNA) maintenance. However, it remains to...Full Text Available

2011-07-01

189

Selective stimulation of catecholamine release from bovine adrenal chromaffin cells by an ionotropic purinergic receptor sensitive to 2-methylthio ATP

Two novel crystal structures of Zea mays protein kinase CK2α catalytic subunit, one in complex with the specific inhibitor 4,5,6,7-tetrabromobenzotriazole (TBB) and another...Full Text Available

2001-11-01

190

Purinergic receptors are involved in tooth-pulp evoked nocifensive behavior and brainstem neuronal activity

Background2-Methylthioadenosine 5'-triphosphate (2-MeSATP), formerly regarded as a specific P2Y (metabotropic) purinergic receptor agonist, stimulates Ca²⁺influx and...Full Text Available

191

P2X1 receptor mobility and trafficking; regulation by receptor insertion and activation

BackgroundTo evaluate whether P2X receptors are involved in responses to noxious pulp stimulation, the P2X₃and P2X_2/3receptor agonist α,β-methyleneATP...Full Text Available

192

NMR of a synthetic peptide spanning the triphosphate binding site of adenosine 5'-triphosphate in actin

P2X1 receptors for ATP contribute to signalling in a variety of cell types and following stimulation undergo rapid desensitisation (within 1 s), and require ∼5 min to recover. In HEK293 cells...Full Text Available

2010-06-01

193

Interaction of N-methyl-2-alkenyl-4-quinolones with ATP-dependent MurE ligase of Mycobacterium tuberculosis: antibacterial activity, molecular docking and inhibition kinetics

The amino acid residues 114-118 in actin were found to be implicated strongly in the binding of nucleotide, and as would be expected for such an important binding site, they are located in a completely conserved region of the actin sequence. A 19-residue peptide with the actin sequence 106-124 was synthesized in order to span the putative triphosphate binding site. Proton NMR spectra of the actin peptide 114-118 in the presence and absence of ATP indicated that Arg-116 and Lys-118 are particularly involved in binding ATP. A strong binding of ATP to the peptide 106-124 also was measured. Tripolyphosphate bound to the peptide 106-124 somewhat more weakly than ATP. Binding involved residues 115-118 and 121-124, indicating the presence of a reverse turn between these segments. Proton resonances were assigned by using two-dimensional double quantum correlated spectroscopy, one-dimensional spin decoupling ...

1987-03-10

194

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

ObjectivesThe aim of this study was to comprehensively evaluate the antibacterial activity and MurE inhibition of a set of N-methyl-2-alkenyl-4-quinolones found...Full Text Available

2011-08-01

195

Expression of Genes Encoding F₁-ATPase Results in Uncoupling of Glycolysis from Biomass Production in Lactococcus lactis

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 this unlabeled analogue ...

1986-01-14

196

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

We studied how the introduction of an additional ATP-consuming reaction affects the metabolic fluxes in Lactococcus lactis. Genes encoding the hydrolytic part of the F₁ domain...Full Text Available

2002-09-01

197

Disruption of the ATP-binding Cassette B7 (ABTM-1/ABCB7) Induces Oxidative Stress and Premature Cell Death in Caenorhabditis elegans*

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

198

Characterization of the major phosphoprotein and its kinase on the surface of the rat adipocyte

X-linked sideroblastic anemia with ataxia (XLSA/A) is a rare inherited disorder characterized by mild anemia and ataxia. XLSA/A is caused by mutations in the ABCB7 gene, which encodes...Full Text Available

2011-06-17

199

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.

Intact rat fat cell exposed to 12.5 ..mu..M (..gamma..-32P)ATP incorporate label into specific proteins within minutes. By solubilizing the reaction mixture with SDS which bypasses the subcellular fractionation steps, the labeled proteins can be identified in autoradiographs of SDS-PAGE gels. The most prominently labeled protein has an M/sub r/ of 42,000. Localization of this component to the cell surface can be made on the basis of inhibition of phosphorylation by addition of a protein derived from the rat brain with protein kinase inhibitory property, susceptibility of the phosphorylated protein to the tryptic digestion, inhibition of phosphorylation of this protein after brief exposure to melittin. To rule out the possibility that the cell surface protein might be a mitochondrial contaminant from broken cells, /sup 32/Pi-labeled and (..gamma..-/sup 32/P)ATP-labeled cells were chromatographed on a rabbit anti-pyruvate dehydrogenase ...

1986-12-01

200

Autocrine regulation of T-cell activation by ATP release and P2X₇ receptors

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

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201

Thymidylate synthase expression and genotype have no major impact on the clinical outcome of colorectal cancer patients treated with 5-fluorouracil

T-cell activation requires the influx of extracellular calcium, although mechanistic details regarding such activation are not fully defined. Here, we show that P2X₇ receptors play a key...Full Text Available

2009-06-01

202

British Library Electronic Table of Contents (United Kingdom)

Background and objectives.Thymidylate synthase (TS) expression levels appear to be related to response to 5-fluorouracil-(5-FU)-based chemotherapy in colorectal cancer (CRC) patients. Three polymorphisms have been proposed as modulators of TS expression: a tandemly repeated sequence (2R/3R) in the 5? UTR, a SNP (G > C) within the 3R allele and a 6 bp deletion in the 3' UTR.To evaluate the influence of TS expression and polymorphisms on clinical outcome of 5-FU-treated patients we performed a comprehensive genetic analysis on 63 CRC patients.Methods.TS expression levels were analyzed in normal and tumor tissues. TS coding sequence and UTR polymorphisms were investigated on DNA from normal tissue. LOH analysis was performed to determine tumor genotype.Results.A difference in disease-free sur...

2011-01-01

203

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

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

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

204

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

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

205

A rapid, radiometric assay for sucrose synthase

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 subunit of Arabidopsis ...

2009-08-17

206

Yeast ribosomal protein L1 is required for the stability of newly synthesized 5S rRNA and the assembly of 60S ribosomal subunits.

Investigations of sucrose synthase in maize root tips have required development of a means to circumvent the rapid decline of activity observed after extraction dialysis and either synthetic or degradative assays. Several protease inhibitors were tested; although PMSF increased initial activity, no inhibitor prevented the drop in activity with time. Western blot analysis indicated that activity decline was not associated with protein degradation. Therefore, a procedure was developed which (1) shortened extraction-to-assay period from ca. 24 hours to 7 minutes, (2) simplified previous assays and (3) reduced the amount of tissue required. Extract was desalted with spun columns and the {sup 14}C-UDPG product recovered with DEAE ion exchange paper. The minute quantities of product recovered can be concealed by the presence of trace impurities in the {sup 14}C-sucrose utilized. DEAE ion exchange paper was used to remove interfering radio-labelled compounds from the {sup ...

1990-05-01

207

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

Ribosomal protein L1 from Saccharomyces cerevisiae binds 5S rRNA and can be released from intact 60S ribosomal subunits as an L1-5S ribonucleoprotein (RNP) particle. To understand the nature of the...Full Text Available

1993-05-01

208

The k43 gene, required for chorion gene amplification and diploid cell chromosome replication, encodes the Drosophila homolog of yeast origin recognition complex subunit 2

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

209

The distribution of active RNA polymerase II along the transcribed region is gene-specific and controlled by elongation factors

Lethal alleles of the Drosophila k43 gene result in small or missing imaginal discs, greatly reduced mitotic index, and fragmented and abnormally condensed chromosomes. A female-sterile...Full Text Available

1997-04-15

210

Specific requirement of the chromatin modifier mSin3B in cell cycle exit and cellular differentiation

In order to study the intragenic profiles of active transcription, we determined the relative levels of active RNA polymerase II present at the 3′- and 5′-ends of 261 yeast genes by...Full Text Available

2010-08-01

211

SWI/SNF and Asf1 Independently Promote Derepression of the DNA Damage Response Genes under Conditions of Replication Stress

The Sin3-histone deacetylase (HDAC) corepressor complex is conserved from yeast to humans. Mammals possess two highly related Sin3 proteins, mSin3A and mSin3B, which serve as scaffolds tethering HDAC...Full Text Available

2008-03-18

212

Primary organization of nucleosomal core particles is invariable in repressed and active nuclei from animal, plant and yeast cells.

The histone chaperone Asf1 and the chromatin remodeler SWI/SNF have been separately implicated in derepression of the DNA damage response (DDR) genes in yeast cells treated with genotoxins that cause...Full Text Available

213

Multiway real-time PCR gene expression profiling in yeast Saccharomyces cerevisiae reveals altered transcriptional response of ADH-genes to glucose stimuli

A refined map for the linear arrangement of histones along DNA in nucleosomal core particles has been determined by DNA-protein crosslinking. On one strand of 145-bp core DNA, histones are aligned in...Full Text Available

1985-05-24

214

BackgroundThe large sensitivity, high reproducibility and essentially unlimited dynamic range of real-time PCR to measure gene expression in complex samples provides the opportunity...Full Text Available

215

Molecular methods

Metabolomic analysis of the plant pathogenic fungi Fusarium graminearum and Fusarium culmorum

Chapter 5, describes some of the most important molecular methods used in the study of chromosome structure and function. The methods discussed include fragmentation of DNA, cloning, flow cytometry and chromosome sorting, is situ hybridization, polymerase chain reaction (PCR), and yeast artificial chromosomes (YACs). 18 refs., 3 figs., 1 tab.

1993-12-31

216

Environmental Research Database

DescriptionThis project is part of the BBSRCs special initiative on plant and microbial metabolomics. The project will primarily focus on the trichothecene mycotoxin producing Ascomycete fungus Fusarium graminearum (Fg) which causes ear blight disease of small grain cereals. The project aims to explore the metabolome of various wild-type and single gene deletion Fg strains and to compare some of these with the identical gene mutation in the budding yeast, S. cerevisiae (Sc) and the saprophytic filamentous [continued...

2008-01-31

217

Hybridization with synthetic oligonucleotides

Genome-wide analysis of N¹-methyl-adenosine modification in human tRNAs

Procedures are described for the use of synthetic oligonucleotides for Southern blot experiments and gene bank screening, and the effect of various mismatches on the efficiency of hybridization is demonstrated. The following topics are discussed: sensitivity vs. specificity, hybridization of a 12-mer to the lambda endolysin gene; hybridization of oligonucleotide probes to the E. coli lac operator; hybridization of synthetic probes to the CYC1 gene of yeast; and cloning eucaryotic genes. (HLW)

1978-01-01

218

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

The N¹-methyl-Adenosine (m¹A58) modification at the conserved nucleotide 58 in the TΨC loop is present in most eukaryotic tRNAs. In yeast, m¹A58 modification...Full Text Available

2010-07-01

219

Enhanced Degradation of an Endocrine-Disrupting Chemical, Butyl Benzyl Phthalate, by Fusarium oxysporum f. sp. pisi Cutinase

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

220

Biological solubilization of coal in aqueous and nonaqueous media

Compared to yeast esterase, fungal cutinase degraded butyl benzyl phthalate (BBP) far more efficiently; i.e., almost 60% of the BBP disappeared within 7.5 h. Also, the final chemical composition significantly...Full Text Available

2002-09-01

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

221

Assembly of 60S ribosomal subunits is perturbed in temperature-sensitive yeast mutants defective in ribosomal protein L16.

Current investigations into the biological solubilization of coal with microorganisms focus on the production of solubilizing activity in fungi. Test organisms for this work include a species of the yeast Candida previously isolated from a lignite outcrop, and P. chrysosporium, a filamentous higher fungus which has played a major role in lignin biodegradation research. The studies described are primarily exploratory in nature, and are fundamental to the design of more sophisticated inquiries into the physiology of fungal coal solubilization.

1987-01-01

222

Single-molecular analysis of the binding state of myosin V and actin

Temperature-sensitive mutants defective in 60S ribosomal subunit protein L16 of Saccharomyces cerevisiae were isolated through hydroxylamine mutagenesis of the RPL16B gene and plasmid shuffling. Two...Full Text Available

1991-11-01

223

Rapid toxicity testing based on mitochondrial respiratory activity

A class-V myosin, myosin V, one of 18 known classes of actin-based motor proteins, plays a role in transporting organelles within a cell. Unlike myosin-II, which functions as an assembly in the thick filaments of muscle, myosin V is a two-headed processive motor protein, which functions as a single molecule: myosin V performs many consecutive steps before it detaches from an actin filament accompanied by catalytic cycles of ATP (adenosine 5'-triphosphate) ase. The mechanism of such chemomechanical steps is explained by a 'Hand-over-hand model' in which two heads of myosin V alternately repeat single-headed and double-headed bindings with an actin filament. To investigate the binding state of myosin V at several key nucleotide states during ATP hydrolysis, we measured the mechanical properties of a single myosin V - actin complex by applying an external load with optical trap.

2006-03-21

224

Radioactive-electrophoretic assay of adenosine 5'-triphosphate sulfurylase activity in crude extracts with sulfate or selenate as a substrate

The need exists for rapid and inexpensive methods to determine the health effects of environmental contaminants on biological systems. One of the current research approaches for assessing cytotoxicity is to monitor the respiratory activity of the mitochondrion, a sensitive, nonspecific subcellular target site. Detected changes in mitochondrial function after the addition of a test chemical could be correlated to toxic effects. Mitochondrial respiration can be characterized by three indices: state 3 and state 4 respiratory rates, and the respiratory control ratio (RCR). State 4, the idle or resting state, results when coupled mitochondrial respire in a medium containing inorganic phosphate and a Kreb's cycle substrate in the absence of a phosphate acceptor such as adenosine diphosphate (ADP). In the presence of ADP the respiration rate increases to a maximum (state 3), accompanied by phosphorylation of ADP to adenosine triphosphate (ATP). The ratio of state ...

1990-05-01

225

Effect of pH on the phase separation in the ternary aqueous system containing the hydrophilic ionic liquid 1-butyl-3-methylimidazolium bromide and the kosmotropic salt potassium citrate at T=298.15K

An assay method for ATP sulfurylase is presented which employs Na/sub 2/(35)SO/sub 4/ as a substrate and measures the production of labeled adenosine 5'-phosphosulfate and 3'-phosphoadenosine 5'-phosphosulfate by low-voltage, hanging paper strip electrophoresis. The method is applicable to crude bacterial or mammalian extracts and accurately measures picomole amounts of product(s). Na/sub 2/(/sup 75/)SeO/sub 4/ can also be employed as a substrate, if the unstable radioactive product, adenosine 5'-phosphoselenate, is converted to elemental /sup 75/Se degrees by inclusion of reduced glutathione in the reaction mixture. The same paper strip electrophoretic technique can then be used to separate /sup 75/Se degrees from the radiolabeled substrate. The method also has utility for measuring any direct reduction by crude microbial extracts of radioactive selenate to selenite, independent of ATP sulfurylase.

1989-02-01

226

British Library Electronic Table of Contents (United Kingdom)

The goal of this work is to study the effect of the aqueous medium pH on the two-phase equilibrium behaviour of the aqueous {1-butyl-3-methylimidazolium bromide ([C4mim]Br)+potassium citrate} system in order to obtain further information about the salting-out effect produced by the addition of a kosmotropic salt to an aqueous solution of a hydrophilic ionic liquid (IL). For this purpose the phase diagrams and the liquid-liquid equilibrium (LLE) data of the {[C4mim]Br+potassium citrate} aqueous two-phase system (ATPS) were determined experimentally at different pH values. Four medium pH values (5.00, 6.00, 7.00 and 8.00) were assayed at T=298.15K. The effect of pH on the ATPS-promoting capability of citrate anions in the aqueous [C4mim]Br solutions was explained based on the structural hydr...

2011-01-01

227

Direct photoaffinity labeling by nucleotides of the apparent catalytic site on the heavy chains of smooth muscle and Acanthamoeba myosins

Photoaffinity labeling of ATP and NAD"+ binding sites on recombinant human interleukin 2

The heavy chains of Acanthamoeba myosins, IA, IB and II, turkey gizzard myosin, and rabbit skeletal muscle myosin subfragment-1 were specifically labeled by radioactive ATP, ADP, and UTP, each of which is a substrate or product of myosin ATPase activity, when irradiated with uv light at 0"0C. With UTP, as much as 0.45 mol/mol of Acanthamoeba myosin IA heavy chain and 1 mol/mol of turkey gizzard myosin heavy chain was incorporated. Evidence that the ligands were associated with the catalytic site included the observations that reaction occurred only with nucleotides that are substrates or products of the ATPase activity; that the reaction was blocked by pyrophosphate which is an inhibitor of the ATPase activity; that ATP was bound as ADP; and that label was probably restricted to a single peptide following limited subtilisin proteolysis of labeled Acanthamoeba myosin IA heavy chain and extensive cleavage with CNBr and trypsin of labeled turkey ...

228

Kinetic, spectroscopic and chemical modification study of iron release from transferrin; iron(III) complexation to adenosine triphosphate

Interleukin 2 (IL-2) is a T-cell-derived lymphokine critical in the activation and proliferation of T cells, B cells, and lymphokine-activated killer cells. It is a glycoprotein of #approx#15,500 daltons that is synthesized and secreted after activation by antigen or mitogen. By using the analogs 8-azidoadensoine 5'-[#gamma#-"3"2P]triphosphate ([#gamma#-"3"2P]8N_3ATP) and nicotinamide 2-azidoadenine [adenylate-"3"2P]dinucleotide ([#alpha#-"3"2P]2N_3NAD"+) as photoaffinity probes, the authors have detected specific, metal ion-requiring nucleotide binding sites on recombinant human IL-2 (rhIL-2). The specificity of these nucleotide interactions with rhIL-2 was demonstrated by saturation effects and by competition by the parent nucleotides at physiologically relevant concentrations. Saturation of photoinsertion into rhIL-2 occurred at 50 #mu#M [#gamma#-"3"2P]8N_3ATP. Saturation of photoinsertion with [#alpha#-"3"2P]2N_3NAD"+ was observed at 180 ...

229

Insulin-like growth factor II receptor is phosphorylated by a tyrosine kinase in adipocyte plasma membranes

Amino acids other than those that serve as ligands have been found to influence the chemical properties of transferrin iron. The catalytic ability of pyrophosphate to mediate transferrin iron release to a terminal acceptor is largely quenched by modification non-liganded histine groups on the protein. The first order rate constants of iron release for several partially histidine modified protein samples were measured. A statistical method was employed to establish that one non-liganded histidine per metal binding domain was responsible for the reduction in rate constant. These results imply that the iron mediated chelator, pyrophosphate, binds directly to a histidine residue on the protein during the iron release process. EPR spectroscopic results are consistent with this interpretation. Kinetic and amino acid sequence studies of ovotransferrin and lactoferrin, in addition to human serum transferrin, have allowed the tentative assignment of His-207 in the N-terminal domain and His-535 ...

1985-01-01

230

Expression of Na(+)-H+ exchange and ATP-dependent proton extrusion in growing rat IMCD cells

Incorporation of /sup 32/P from (gamma-32P)ATP into tyrosine residues of the insulin-like growth factor (IGF)-II receptor was observed in a Triton X-100-insoluble fraction of rat adipocyte plasma membranes. IGF-II receptor phosphorylation proceeded to a stoichiometry of approximately 0.5 mol of phosphate/IGF-II binding site after 10 min of incubation at 4 degrees C. A Km for ATP of 6 microM was calculated for this phosphorylation reaction. Addition of IGF-II caused an approximately 2-fold increase in tyrosine phosphorylation of the IGF-II receptor in this preparation. In contrast, phosphorylation of angiotensin II by the Triton X-100 washed membranes was not stimulated by IGF-II. Incubation of purified receptor immobilized on IGF-II agarose or of receptor-enriched low density microsomal membranes with (gamma-32P)ATP did not result in appreciable incorporation of (/sup 32/P)phosphate into the IGF-II receptor nor into ...

1986-06-15

231

ATP-dependent partitioning of the DNA template into supercoiled domains by Escherichia coli UvrAB

As the last step of urinary acidification, the inner medullary collecting duct (IMCD) is thought to secrete protons into the tubular lumens by means of a H(+)-translocating adenosinetriphosphatase (H(+)-ATPase). However, recent studies have also shown the existence of Na(+)-H+ exchange activity in IMCD cells. Although the physiological function of the antiporter in IMCD cells is unknown, activation of Na(+)-H+ exchange in other cell-culture systems has been suggested to be closely associated with the process of cell growth. Thus presence of Na(+)-H+ exchange may relate to the growth phase of these cells. To examine intracellular pH (pHi) regulation in growing IMCD cells, we studied proton transport by Na(+)-dependent and Na(+)-independent mechanisms by microfluorimetry using the pHi-sensitive dye 2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein acetoxymethyl ester (BCECF/AM). Actively growing cells, defined by (3H)thymidine incorporations, demonstrated an ...

1990-03-01

232

Mutations in cyr1 and pat1 reveal pheromone-induced G1 arrest in the fission yeast Schizosaccharomyces pombe

The helicase action of the Escherichia coli UvrAB complex on a covalently closed circular DNA template was monitored using bacterial DNA topoisomerase I, which specifically removes negative supercoils. In the presence of E. coli DNA topoisomerase I and ATP, the UvrAB complex gradually introduced positive supercoils into the input relaxed plasmid DNA template. Positive supercoils were not produced when E. coli DNA topoisomerase I was replaced by eukaryotic DNA topoisomerase I or when both E. coli and eukaryotic DNA topoisomerases I were added simultaneously. These results suggest that like other DNA helix-tracking processes, the ATP-dependent action of the UvrAM complex on duplex DNA simultaneously generates both positive and negative supercoils, which are not constrained by protein binding but are torsionally strained. The supercoiling activity of UvrAB on UV-damaged DNA was also studied using UV-damaged plasmid DNA and a mutant UvrA protein ...

233

DEFF Research Database (Denmark)

Investigations into sexual differentiation and pheromone response in the fission yeast Schizosaccharomyces pombe are complicated by the need to first starve the cells of nitrogen. Most mating-related experiments are therefore performed on non-dividing cells. Here we overcome this problem by using two mutants that bypass the nutritional requirements and respond to the M-factor mating pheromone in rich medium. The first mutant lacks the cyr1 gene which encodes adenylate cyclase and these cells contain no measurable amounts of cAMP. When M-factor is added to a growing h+ cyr1- strain it causes a transient G1 arrest of cell division, transcription of mat1-Pm, and elongation of the cells to form shmoos. The second mutant contains the temperature-sensitive pat1-114 allele. At 30 degrees C this mutant was previously shown not only to bypass the nutritional signal but also to stop growing in a state derepressed for pheromone-controlled functions. We now report that an h+ ...

1994-01-01

234

Mitochondrial genetic damage induced in yeast by a photoactivated furocoumarin in combination with ethidium bromide or ultraviolet light

Management of industrial solid wastes in Alexandria, Egypt

Ethidium bromide (EB) and ultraviolet light (UV) in combination are known to produce a synergistic induction of 'petite' mutants in yeast. Two other agents were combined with EB, 3-Carbethoxypsoralene (3 CPs) activated by 365 nm light or #gamma# rays. EB in combination with 3 CPs also resulted in an enhanced production of 'petite' mutants. After the photoaddition of 3 CPs in exponential phase cells, recovery of the 'petite' mutation during dark liquid holding was inhibited by the presence of EB producing an enhanced number of 'petite' mutants. The behavior of mitochondrial antibiotic resistance markers after individual and combined treatments with EB and 3 CPs indicates a random loss of markers after EB and a preferential loss of a certain region for the 3 CPs photoaddition. The combination of the two agents leads to an additivity of total drug marker losses rather than a synergistic loss. The combination of EB with #gamma# rays produced no enhancement in 'petite' ...

235

Mineral biotechnology. Microbial aspects of mineral beneficiation, metal extraction, and environmental control

This paper presents a summary of the first phase of the EPA project, which encompasses surveys of residues from industrial sources in Alexandria. Studies to date indicate that wastes from various industries can be recovered economically. Wastes such as tin cans, glass, wastepaper, and food residues from processing of fruits, starch, and beer are examples of reusable industrial wastes in Egypt. The results of experimental studies for reuse of residues from oil refining, starch and yeast processing, and steel pickling are presented. Spent clay from edible oil refining is currently discarded, causing both handling and disposal problems. This clay contains as much as 40% oil; 90% can be recovered by extraction. The recovered oil can be successfully used in soap production, and the spent clay can be reused in oil bleaching. Other examples include starch and yeast wastes, which can be used for animal feed, and spent pickling liquor, which can be used ...

1983-03-01

236

Mechanism of biodegradation of paraquat by Lipomyces starkeyi

Papers in this book illustrate the utility of mineral biotechnology with respect to biobeneficiation, bioleaching, bioremediation and biomineralization. Papers of particular interest to the coal industry include: depression of pyrite flotation by yeast and bacteris (S.K. Kawatra and T.C. Eisele); desulfurization of coal by microbial flotation in a semicontinuous system (T. Nagaoka and others); biochemical removal of HAP precursors from coal - INEEL slurry column testing (K.S. Noah and G.J. Olson); microorganisms, biotechnology and acid rock drainage - emphasis on passive-biological control and treatment methods (N. Kuyucak); and utility of bioreagents in mineral processing (P. Somasundaran and others).

2001-07-01

237

Mechanism of biodegradation of paraquat by Lipomyces starkeyi

The biodegradation of ring-/sup 14/C- and methyl-/sup 14/C-labeled paraquat by the soil yeast Lipomyces starkeyi was studied in vitro. It was found that the degradation of paraquat (acting as a sole source of culture nitrogen) resulted in the accumulation in the extracellular medium of radiolabeled acetic acid. The culture also evolved radiolabeled CO/sub 2/. The results suggest that the degradation of paraquat by L. starkeyi is associated with the integrity of the cell wall and that disruption or removal of the wall results in a complete loss of degradative capability. A mechanism for the degradation of paraquat by this organism is postulated.

1985-05-01

238

Latvian scientists research into chemical uses of timber

The biodegradation of ring-"1"4C- and methyl-"1"4C-labeled paraquat by the soil yeast Lipomyces starkeyi was studied in vitro. It was found that the degradation of paraquat (acting as a sole source of culture nitrogen) resulted in the accumulation in the extracellular medium of radiolabeled acetic acid. The culture also evolved radiolabeled CO_2. The results suggest that the degradation of paraquat by L. starkeyi is associated with the integrity of the cell wall and that disruption or removal of the wall results in a complete loss of degradative capability. A mechanism for the degradation of paraquat by this organism is postulated.

239

Antifungal activity of Glycyrrhiza glabra extracts and its active constituent glabridin

Scientists of the Institute of Wood Chemistry of the Latvian Academy of Sciences have developed two highly efficient processes for producing furfural, a feedstock for varnishes, synthetic resins and plastics. It is made of production wastes, including branches and small-dimension timber. By one process, the raw material is chipped, treated first with diluted sulphuric acid and then with steam heated to 250 degrees C. The other uses concentrated sulphuric acid as a catalyst. Besides furfural, this process also yields sugar solutions used in alcohol and nutrient yeast production.

1982-03-20

240

British Library Electronic Table of Contents (United Kingdom)

Glabridin, an active constituent of Glycyrrhiza glabra roots, was found to be active against both yeast and filamentous fungi. Glabridin also showed resistance modifying activity against drug resistant mutants of Candida albicans at a minimum inhibitory concentration of 31.25-250 g/mL. Although the compound was reported earlier to be active against Candida albicans, but this is the first report of its activity against drug resistant mutants. Copyright Copyright 2009 John Wiley & Sons, Ltd.

2009-01-01

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

241

Antifungal activity of the extracts and neolignans from Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck

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

Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck (Piperaceae) is a medicinal plant traditionally used in Brazil to treat infectious diseases. The extracts obtained from the leaves of P. regnellii were investigated for their antifungal activities against the yeasts Candida albicans, C. krusei, C. parapsilosis, and C. tropicalis. The EtOAc extract presented gnificant activity against Candida albicans with MIC at 125 {mu}g mL{sup -1}, and a moderate activity against both C. krusei and C. parapsilosis with MIC at 500 {mu}g mL{sup -1}. Candida tropicalis was not inhibited by this extract at concentrations as high as 1000 {mu}g mL{sup -1}. Based on these findings, the EtOAc extract was fractionated by silica gel column chromatography into nine fractions. The hexane and CHCl{sub 3} fractions showed varied levels of antifungal activity against all test yeasts. Further column chromatography separation of the hexane fraction afforded the pure ...

2005-11-15

242

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

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

243

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

244

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

245

Isoflavone content and apoptotic effect in HT-29 cancer cells of a soy germ extract

British Library Electronic Table of Contents (United Kingdom)

Colon cancer is one of the leading causes of death and a major public health problem in western countries. We examined the isoflavone content of 70% ethanol extract of soy germ (SG) and its apoptotic effect in HT-29 colon cancer cells. Our results showed that the major isoflavones of the SG extract were daidzein and genistein, and it effectively induced apoptosis in HT-29 cancer cells. In addition, nuclear factor-kappa B (NF-kB) expression was reduced in cells treated with the SG extract, which reduced cyclooxygenase-2 (COX-2) and nitric oxide synthase (iNOS) expression. These combined effects ultimately resulted in apoptosis via caspase-3 activation. In addition, daidzein and genistein, the major isoflavones of SG extract, also exerted the apoptotic effect against HT-29 cancer cells. Thes...

2012-01-01

246

Effects of Dietary Glycyrrhizin on Growth and Nonspecific Immunity of White Shrimp, Litopenaeus vannamei

British Library Electronic Table of Contents (United Kingdom)

The growth response, total hemocyte count (THC), respiratory burst (release of superoxide anion), phenoloxidase (PO) activity, superoxide dismutase (SOD) activity, nitric oxide synthase (NOS) activity as well as resistance to the pathogen Vibrio alginolyticus were measured in Litopenaeus vannamei, which had been fed diets supplemented with glycyrrhizin (the aqueous extract of licorice, Glycyrrhiza glabra, roots) at 0, 50, 100, 150, and 200 mg/kg of feed for 8 wk. In the feeding trial, there was no significant difference in survival rate (P > 0.05). Significant higher specific growth rate was observed in treatments with dietary glycyrrhizin than that in the control group (P < 0.05). Shrimp fed a diet with 200 mg/kg of glycyrrhizin had significant higher THC, PO activity, respiratory burst a...

2010-01-01

247

Distribution of NADPH-diaphorase activity in the central nervous system of the young and adult land snail Megalobulimus abbreviatus

British Library Electronic Table of Contents (United Kingdom)

Nitric oxide (NO) is a gas produced through the action of nitric oxide synthase that acts as a neurotransmitter in the central nervous system (CNS) of adult gastropod mollusks. There are no known reports of the presence of NOS-containing neurons and glial cells in young and adult Megalobulimus abbreviatus. Therefore, NADPH-d histochemistry was employed to map the nitrergic distribution in the CNS of young and adult snails in an attempt to identify any transient enzymatic activity in the developing CNS. Reaction was observed in neurons and fibers in all CNS ganglia of both age groups, but in the pedal and cerebral ganglia, positive neurons were more intense than in other ganglia, forming clusters symmetrically located in both paired ganglia. However, neuronal NADPH-d activity in the mesocer...

2010-01-01

248

ALK, the Key Gene for Gelatinization Temperature, is a Modifier Gene for Gel Consistency in Rice

British Library Electronic Table of Contents (United Kingdom)

Abstract Gelatinization temperature (GT) is an important parameter in evaluating the cooking and eating quality of rice. Indeed, the phenotype, biochemistry and inheritance of GT have been widely studied in recent times. Previous map-based cloning revealed that GT was controlled by ALK gene, which encodes a putative soluble starch synthase II-3. Complementation vector and RNAi vector were constructed and transformed into Nipponbare mediated by Agrobacterium. Phenotypic and molecular analyses of transgenic lines provided direct evidence for ALK as a key gene for GT. Meanwhile, amylose content, gel consistency and pasting properties were also affected in transgenic lines. Two of four nonsynonymous single nucleotide polymorphisms in coding sequence of ALK were identified as essential for GT. ...

2011-01-01

249

A Hypothesis: Supplementation with Mushroom-Derived Active Compound Modulates Immunity and Increases Survival in Response to Influenza Virus (H1N1) Infection.

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

We hypothesize that the mushroom-derived active compound may be a potential strategy for increasing survival in response to influenza virus (H1N1) infection through the stimulation of host innate immune response. The validity of the hypothesis can be tested by immune response to influenza infection as seen through survival percentage, virus clearance, weight loss, natural killer cell cytotoxicity, Tumor Necrosis Factor-? (TNF-?) and Interferon-gamma (IFN-?) levels, lytic efficiency in the spleens of mice and inducible nitric oxide synthase mRNA expressions in RAW 264.7 murine macrophage cells. The hypothesis may improve people's quality of life, reduce the medical cost of our healthcare system and eliminate people's fears of influenza outbreak. PMID:21660092

2011-03-20

250

Coordinated responses of phytochelatin synthase and metallothionein genes in black mangrove, Avicennia germinans, exposed to cadmium and copper

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 ...

2010-03-08

251

Coordinated responses of phytochelatin synthase and metallothionein genes in black mangrove, Avicennia germinans, exposed to cadmium and copper

To evaluate the role of phytochelatins and metallothioneins in heavy metal tolerance of black mangrove Avicennia germinans, 3-month-old seedlings were exposed to cadmium or copper for 30 h, under hydroponic conditions. Degenerate Mt2 and PCS primers were synthesized based on amino acid and nucleotide alignment sequences reported for Mt2 and PCS in other plant species found in GenBank. Total RNA was isolated from A. germinans leaves and two partial fragments of metallothionein and phytochelatin synthase genes were isolated. Gene expression was evaluated with reverse transcripatase-polymerase chain reaction (RT-PCR) amplification technique. Temporal analysis showed that low Cd{sup 2+} and Cu{sup 2+} concentrations caused a slight (but not significant) increase in AvMt2 expression after a 16 h exposure time, while AvPCS expression showed a significant increase under the same conditions but only after 4 h. Results strongly suggest that the rapid increase in AvPCS ...

2007-08-01

252

Biosynthesis of sucrose and mannitol as a function of leaf age in celery (Apium graveolens L. )

To evaluate the role of phytochelatins and metallothioneins in heavy metal tolerance of black mangrove Avicennia germinans, 3-month-old seedlings were exposed to cadmium or copper for 30 h, under hydroponic conditions. Degenerate Mt2 and PCS primers were synthesized based on amino acid and nucleotide alignment sequences reported for Mt2 and PCS in other plant species found in GenBank. Total RNA was isolated from A. germinans leaves and two partial fragments of metallothionein and phytochelatin synthase genes were isolated. Gene expression was evaluated with reverse transcripatase-polymerase chain reaction (RT-PCR) amplification technique. Temporal analysis showed that low Cd"2"+ and Cu"2"+ concentrations caused a slight (but not significant) increase in AvMt2 expression after a 16 h exposure time, while AvPCS expression showed a significant increase under the same conditions but only after 4 h. Results strongly suggest that the rapid increase in AvPCS expression ...

2007-08-01

253

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

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 ...

1988-01-01

254

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

255

Some sulfonamide drugs inhibit ATPase activity of heat shock protein 90: investigation by docking simulation and experimental validation

British Library Electronic Table of Contents (United Kingdom)

Eight selected sulfonamide drugs were investigated as inhibitors of heat shock protein 90 (Hsp90). The investigation included simulated docking experiments to fit the selected compounds within the binding pocket of Hsp90. The selected molecules were found to readily fit within the ATP-binding pocket of Hsp90 in low-energy poses. The sulfonamides torsemide, sulfathiazole, and sulfadiazine were found to inhibit the ATPase activity of Hsp90 with IC50 values of 1.0, 2.6, and 1.5 mM, respectively. Our results suggest that these well-established sulfonamides can be good leads for subsequent optimization into potent Hsp90 inhibitors.

2011-01-01

256

Nerve-Evoked Constriction of Rat Tail Veins Is Potentiated and Venous Diameter Is Reduced after Chronic Spinal Cord Transection

British Library Electronic Table of Contents (United Kingdom)

Abstract Despite reduced sympathetic activity below the level of a spinal cord injury (SCI), venoconstriction during autonomic dysreflexia increases venous return to the heart. Here, contractions of isometrically mounted tail veins from rats with spinal transection at T4 performed 8???10 weeks earlier are compared with those from sham-operated rats. After SCI, lumen diameter was reduced by ?30% and the contractions evoked by electrical stimulation of the perivascular axons were larger than control. This augmentation of neurovascular transmission was not associated with enhanced sensitivity to ?-adrenoceptor agonists or to adenosine-5?-triphosphate (ATP) although contractions to depolarization with K+ were larger after SCI. The percentage reduction in nerve-evoked contraction after SCI prod...

2011-01-01

257

Mitochondria-localized NAD biosynthesis by nicotinamide mononucleotide adenylyltransferase in Jerusalem artichoke (Helianthus tuberosus L.) heterotrophic tissues

British Library Electronic Table of Contents (United Kingdom)

Current studies in plants suggest that the content of the coenzyme NAD is variable and potentially important in determining cell fate. In cases that implicate NAD consumption, re-synthesis must occur to maintain dinucleotide pools. Despite information on the pathways involved in NAD synthesis in plants, the existence of a mitochondrial nicotinamide mononucleotide adenylyltransferase (NMNAT) activity which catalyses NAD synthesis from nicotinamide mononucleotide (NMN) and ATP has not been reported. To verify the latter assumed pathway, experiments with purified and bioenergetically active mitochondria prepared from tubers of Jerusalem artichoke (Helianthus tuberosus L.) were performed. To determine whether NAD biosynthesis might occur, NMN was added to Jerusalem artichoke mitochondria (JAM)...

2011-01-01

258

Mechanisms of Reproductive Thermotolerance in Gossypium hirsutum: The Effect of Genotype and Exogenous Calcium Application

British Library Electronic Table of Contents (United Kingdom)

Abstract Although photosynthetic thermotolerance has been investigated extensively in cotton leaves, reports on the biochemical influence of the pistil in promoting fertilization thermostability are limited. To evaluate the effect of temperature, genotype, and exogenous calcium application on fertilization and pistil biochemistry in cotton, thermosensitive (cv. ST4554 B2RF) and thermotolerant (cv. VH260) plants were grown under control (30/20-C) or high-temperature (38/20-C) conditions during flowering, and exogenous CaCl2 was applied to flowers 1-day prior to anthesis. Measured pistil parameters included fertilization efficiency; protein concentration; glutathione reductase, superoxide dismutase (SOD) and NADPH oxidase activities; and ATP and calcium levels. Exogenous calcium had no effec...

2011-01-01

259

Impaired mitochondrial energy metabolism and kinetic properties of cytochrome oxidase following acute aluminium phosphide exposure in rat liver

British Library Electronic Table of Contents (United Kingdom)

The present study was designed with an aim to analyze the effect of acute aluminium phosphide (ALP) exposure (10mg/kg b.wt, intragastrically) on the kinetic characteristics of cytochrome oxidase and energy metabolism in male Wistar rat liver mitochondria. Liver mitochondrial preparations from ALP-treated rats exhibited significant decrease (66%) in the activity of cytochrome oxidase suggesting that there was a decrease in the catalytic efficiency of the active oxidase molecules on ALP treatment. The decreased activity of cytochrome oxidase with altered NADH and succinic dehydrogenase activities might have contributed towards a significant decline in state 3 and state 4 respiration as observed. These alterations in the electron transport chain complexes in turn adversely affected the ATP sy...

2010-01-01

260

Fluorocarbons and cardiac arrhythmia: does difluorodichloromethane (FC 12) inhibit cardiac metabolism

Detrital processing in streams exposed to acidic precipitation in the Central Appalachian Mountains

Certain fluorocarbons, such as difluorodichloromethane (FC 12), depress the cardiovascular system by diminution of all the transmembrane ionic conductances in cardiac tissues. Does FC 12 also inhibit active transport and thus enzymatic activity and cellular energy. We measured phosphocreatine (PC), adenosine triphosphate (ATP) and cyclic adenosine monophosphate (AMPc) in rat hearts. Rats were randomly divided into 4 groups; 2 control groups: one breathing a mixture of oxygen (21%) and nitrogen (79%) (group C) and the other breathing the same mixture but simultaneously perfused with 1 microgram/kg/min. epinephrine (groupe E-C); 2 trial groups T and E-T where nitrogen was replaced by FC 12. The maximal FC 12 concentration of 720 micrograms/ml in arterial blood produced no significant difference in the concentrations of these three metabolites compared with controls.

1986-01-01

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

261

Conjugation is essential for the anticholestatic effect of NorUrsodeoxycholic acid in taurolithocholic acid induced cholestasis in rat liver

Continuing high rates of acidic deposition in the eastern United States may lead to long-term effects on stream communities, because sensitive catchments are continuing to lose anions and cations. A two-year study of the effects of pH and associated water chemistry variables on detrital processing in three streams with different bedrock geology in the Monongahela National Forest, West Virginia were investigated. Leaf pack processing rates and macroinvertebrate colonization and microbial biomass (ATP concentration) on the packs in the three stream were compared. It was found that macroinvertebrate and microbial communities differed both among streams that differed in their capacity to buffer the effects of acidic precipitation and among years in the same stream; these differences in biotic communities were not large enough to affect rates of leaf processing between the two years of the study, but they did significantly affect processing rates between acidic and ...

262

British Library Electronic Table of Contents (United Kingdom)

Abstract NorUDCA (24 norursodeoxycholic acid), the C23 homolog of ursodeoxycholic acid (UDCA), showed remarkable therapeutic effects in cholestatic Mdr2 (Abcb4) (multidrug resistance protein 2/ATP binding cassette b4) knockout mice with sclerosing/fibrosing cholangitis. In contrast to UDCA, norUDCA is inefficiently conjugated in human and rodent liver, and conjugation has been discussed as a key step for the anticholestatic action of UDCA in cholestasis. We compared the choleretic, anticholestatic, and antiapoptotic properties of unconjugated and taurine conjugated UDCA (C24) and norUDCA (C23) in isolated perfused rat liver (IPRL) and in natrium/taurocholate cotransporting polypeptide (Ntcp) transfected human hepatoma (HepG2) cells. Taurolithocholic acid (TLCA) was used to induce a predomi...

2010-01-01

263

Analysis of even harmonics generation in an isolated electric power system

British Library Electronic Table of Contents (United Kingdom)

Harmonics bred from loads are mainly odd order because the current waveforms have half-wave symmetry. Since the even harmonics are negligibly small, they generally are not measured in electric power systems. However, even harmonics were measured at a 500/275/154 kV substation in Hokuriku Electric Power Company after a transmission line fault was corrected. The even harmonics caused malfunctions of protective digital relays because the relays used 4th harmonics at the input filter as the automatic supervisory signal. This paper describes mechanisms of generation of the even harmonics by comparing the measured waveforms with the ATP-EMTP simulation results. Analysis of these results has clearly shown that there are three different mechanisms of generation of even harmonics. The first mechani...

2009-01-01

264

A PAC containing the human mitochondrial DNA polymerase gamma gene (POLG) maps to chromosome 15q25

Wiley::The Wiley Encyclopedia of Packaging Technology, 3rd Edition

The human mitochondrial DNA (mtDNA) is a closed circular, 16,569-bp double-stranded DNA, encoding 13 genes whose protein products are subunits of the oxidative phosphorylation system required for synthesis of most of the ATP consumed by eukaryotic cells. Point mutations of the mtDNA that cause multi-tissue, loss-of-energy syndromes, called mitochondrial encephalomyopathies (e.g., MERRF and MELAS), have been identified. In addition, large-scale deletions of the human mtDNA have been identified and are the molecular bases for the neonatal and adolescent onset loss-of-energy syndromes Pearson and Kearns-Sayer, respectively. 5 refs., 1 fig.

1997-03-01

265

Wastenet

...The+Wiley+Encyclopedia+of+Packaging+Technology%2C+3rd+Edition FO21+Food+Packaging Wiley::The Wiley Encyclopedia of Packaging Technology, 3rd Edition WILEY ...US | HELP Home / Chemistry / Food Science & Technology / Food Processing, Production & Manufacture / Food Packaging / The Wiley Encyclopedia of Packaging Technology, 3rd Edition ...Manufacture Sensory Science Food Biotechnology Food Chemistry Food Engineering Related Titles Food Packaging Brewing Yeast and Fermentation by Chris Boulton, David Quain Food ...Editor) Journal of Food Process Engineering Journal of Food Processing and Preservation Packaging Research in Food Product Design and Development by Howard R. ...

266

The antimicrobial efficacy of a silver alginate dressing against a broad spectrum of clinically relevant wound isolates

British Library Electronic Table of Contents (United Kingdom)

Wound dressings impregnated with silver have a role to play in aiding to reduce both the dressing and wound microbial bioburden. It is therefore imperative that antimicrobial wound dressings have efficacy on a broad range of clinical significant microorganisms. Accordingly, this study aimed to determine the antimicrobial efficacy of a silver alginate dressing against 115 wound isolates that had been isolated routinely from patients at West Virginia University Hospital. Standardised corrected zones of inhibition (CZOIs) were performed on all clinical isolates. It was found that the silver alginate dressing was able to inhibit the growth of all microorganisms tested. In particular, the silver alginate dressing inhibited the growth of Candida albicans and yeasts with CZOI of 3-115 mm. All met...

2011-01-01

267

The DFNA5 gene, responsible for hearing loss and involved in cancer, encodes a novel apoptosis-inducing protein

British Library Electronic Table of Contents (United Kingdom)

DFNA5 was first identified as a gene causing autosomal dominant hearing loss (HL). Different mutations have been found, all exerting a highly specific gain-of-function effect, in which skipping of exon 8 causes the HL. Later reports revealed the involvement of the gene in different types of cancer. Epigenetic silencing of DFNA5 in a large percentage of gastric, colorectal and breast tumors and p53-dependent transcriptional activity have been reported, concluding that DFNA5 acts as a tumor suppressor gene in different frequent types of cancer. Despite these data, the molecular function of DFNA5 has not been investigated properly. Previous transfection studies with mutant DFNA5 in yeast and in mammalian cells showed a toxic effect of the mutant protein, which was not seen after transfection ...

2011-01-01

268

Synthesis and study of the antifungal activity of new mono- and disubstituted derivatives of a genetically engineered polyene antibiotic 28,29-didehydronystatin A1 (S44HP)

British Library Electronic Table of Contents (United Kingdom)

Mono- and disubstituted novel derivatives of the heptaene nystatin analog 28,29-didehydronystatin A1 (S44HP, 1) were obtained by chemical modification of the exocyclic C-16 carboxyl and/or an amino group of mycosamine moiety. The strategy of preparation of mono- and double-modified polyene macrolides was based on the use of intermediate hydrophobic N-Fmoc (9-fluorenylmethoxycarbonyl) derivatives that facilitated the procedures of isolation and purification of new compounds. The antifungal activity of the new derivatives was first tested in vitro against yeasts and filamentous fungi, allowing the selection of the most active compounds that were subsequently tested for acute toxicity in mice. 2-(N,N-dimethylamino)ethylamide of 1 (2) and 2-(N,N-dimethylamino)ethylamide of N-fructopyranosyl-28...

2010-01-01

269

Loss of cell components during rehydration of dried Rhodotorula glutinis and its implications for lead uptake

British Library Electronic Table of Contents (United Kingdom)

Abstract Microbial cells are routinely dried and ground before they are used in metal biosorption studies. In this work, a metal biosorbent was prepared by drying biomass of the yeast Rhodotorula glutinis in an oven at 70C for 24-h followed by grinding. Two forms of the prepared biosorbent particles, washed and unwashed, were examined for their ability to remove lead from solution. It was found that the unwashed biosorbent exhibited higher lead uptake than the washed biosorbent. Analysis of the supernatant of washed cells incubated in water and that of unwashed cells incubated in lead solution revealed the presence of protein, carbohydrates, organic acids and inorganic phosphate. Overall, the washed and unwashed cells leached, respectively, 14.5 and 13.4% of their initial dry weight (100-m...

2011-01-01

270

In vitro effects of medium tonicity, nutrient concentration, and free chlorine content on Acanthamoeba

British Library Electronic Table of Contents (United Kingdom)

Background: The environment preferred by Acanthamoeba trophozoites and the mechanism by which the amebae enters the cornea are not yet fully understood. A better understanding of the pathogenesis of this disease may help with prevention and treatment. Purpose: To define the preferred environments for Acanthamoeba survival and proliferation in vitro by examining the effect of tonicity, nutrient concentration, and free chlorine content on Acanthamoeba. Materials and methods: Human corneal isolates of Acanthamoeba castellanii and Acanthamoeba polyphaga trophozoites were cultured at 22^oC (room temperature) in PYG (peptone-yeast extract-glucose) medium. The effect of tonicity on amebae was determined by incubating trophozoites in sodium chloride solutions in concentrations ranging from 0% to 1...

2011-01-01

271

Human type I pituitary adenylate cyclase activating polypeptide receptor (ADCYAP1R): Localization to chromosome band 7p14 and integration into the cytogenetic, physical, and genetic map of chromosome 7

Enhancement of bioleaching of a spent Ni/Mo hydroprocessing catalyst by Penicillium simplicissimum

The gene encoding the human type I pituitary adenylate cyclase activating polypeptide receptor (ADCYAP1R1) was mapped to chromosome 7 by PCR analysis of genomic DNA from a human/rodent somatic cell hybrid mapping panel. This assignment was confirmed and the gene localized to chromosome band 7p14 by fluorescence in situ hybridization. A yeast artificial chromosome containing ADCYAP1R1 was identified in the CEPH {open_quotes}B{close_quotes} Mega-YAC library. This YAC includes two highly polymorphic dinucleotide repeat sequences that will facilitate genetic studies of the contribution of ADCYAP1R1 in disease states of the central nervous and neuroendocrine systems. 13 refs., 1 fig.

1994-10-01

272

British Library Electronic Table of Contents (United Kingdom)

Statistically based experimental designs were applied to screen and optimize the bioleaching of spent hydrocracking catalyst by Penicillium simplicissimum. Eleven factors were examined for their significance on bioleaching using a Plackett-Burman factorial design. Four significant variables (pulp density, sucrose, NaNO"3, and yeast extract concentrations) were selected for the optimization studies. The combined effect of these variables on metal bioleaching was studied using a central composite design (CCD). Second-order polynomials were established to identify the relationship between the recovery percent of the metals and the four significant variables. The optimal values of the variables for maximum metals bioleaching were as follows: pulp density (4.0%, w/v), sucrose (90g/L), NaNO"3 (2...

2011-01-01

273

Curcumin Binding to DNA and RNA

British Library Electronic Table of Contents (United Kingdom)

Curcumin, the yellow pigment from the rhizoma of Curcuma longa, is a widely studied phytochemical with a variety of biological activities. The ongoing research and clinical trials have proved that this natural phenolic compound has great and diverse pharmacological potencies. Beside its effective antioxidant, antiinflammatory, and antimicrobial/antiviral properties, curcumin is also considered as a cancer chemopreventive agent. While the antioxidant activity of curcumin is well documented, its interaction with DNA and RNA is not fully investigated. This study was designed to examine the interactions of curcumin with calf thymus DNA and yeast RNA in aqueous solution at physiological conditions, using constant DNA and RNA concentration (6.25?mM) and various curcumin/polynucleotide (phosphate...

2009-01-01

274

Comparison of media and methods for detecting and enumerating Listeria monocytogenes in refrigerated cabbage.

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

Direct plating, selective enrichment, and cold enrichment followed by secondary selective enrichment procedures were compared for detecting and enumerating Listeria monocytogenes in chopped cabbage stored at 5 degrees C for up to 64 days. Addition of Fe3+ to solid media enhanced detection of the organism. Cold enrichment (5 degrees C) in nutrient broth and brain heart infusion broth followed by secondary enrichment (48 h, 30 degrees C) in Trypticase soy-yeast extract-antibiotic broth and thiocyanate-nalidixic acid broth and plating on selective agar media (Doyle and Schoeni selective enrichment agar [minus acriflavin hydrochloride, supplemented with 5 micrograms of Fe3+/ml] and McBride Listeria agar) resulted in the detection of highest populations. PMID:3111369

1987-05-01

275

Biological treatment of wine of distilleries

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

276

Bioconvertion of spent cellulose sausage casings

The potential of the yeast Candida tropicalis and Candida guillermondii was evaluated and an isolated partnership of microorganisms of waters of the Medellin River, conformed by two bacteria and one leavening, to degrade the content of organic matter present in wine produced by the factory of Licores and Alcoholes de Antioquia (FLA) in aerobic process with biomass production. For each one of the microorganisms in study this capacity of removal in units of chemical demand of oxygen was quantified (CDO); in addition, parameters were analyzed such as yield of the biomass in relation to the removed CDO and to total reducing sugars (TRS) consumed, time of fermentation and speed of growth different dilutions from wine. Also the possible inhibition was analyzed that the present phenolic compounds in this wine can cause in the biological process of degradation.

277

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

278

Antimicrobial silver-montmorillonite nanoparticles to prolong the shelf life of fresh fruit salad

British Library Electronic Table of Contents (United Kingdom)

In this work, silver-montmorillonite (Ag-MMT) antimicrobial nanoparticles have been obtained by allowing silver ions from nitrate solutions to replace the Na^+ of natural montmorillonite and then to be reduced by a thermal treatment. Ag-MMT were used as active antimicrobial compounds to improve the shelf life of fresh fruit salad. In order to assess their influence on product shelf life, sensorial and microbiological quality has been monitored during the storage. The microbiological quality was determined by monitoring the principal spoilage microorganisms (mesophilic and psychrotrophic bacteria, coliforms, lactic acid bacteria, yeasts and molds). Additionally, the evolution of sensorial quality was assessed by monitoring color, odor, firmness and product overall quality. The Ag-MMT nanopa...

2011-01-01

279

Amphiphysin (Amph) maps to the proximal region of mouse chromosome 13

Aerobic Decolorization and Detoxification of a Disperse Dye in Textile Effluent by a New Isolate of Bacillus sp.

Amphiphysin is a protein concentrated in neuronal synapses and peripherally associated with neurotransmitter vesicles. It is expressed in many neurons of the central and peripheral nervous systems, in the adrenal medulla, in the anterior and posterior pituitary, in cell lines of the endocrine pancreas, and in spermatocytes. Its subcellular localization and tissue distribution indicate a potential involvement in mechanisms of regulated exocytosis. A role in the dynamic organization of the membrane-associated cytoskeleton is suggested by structural homology to the products of two yeast genes, RVS161 and RVS167, whose mutation results in an abnormal actin distribution, disturbs budding morphology, and impairs cell entry into stationary phase. Limited stretches of sequence similarity, including an SH3 domain, are also shared with other actin-binding proteins. Amphiphysin is the dominant autoantigen in paraneoplastic Stiff-Man syndrome, a neurological autoimmune ...

1995-07-20

280

British Library Electronic Table of Contents (United Kingdom)

A number of aerobic species capable of decolorizing some of the dyes in a textile mill effluent were isolated. One of the isolates was able to decolorize Terasil black dye under aerobic conditions in the presence of an exogenous carbon source after 5 days. Glucose or starch (%1 ea) are essential for decolorization but the process proceeds faster in the presence of 0.5% yeast extract. Results of the BOD5 show that the untreated effluent samples have a low BOD value, whereas treated samples show an initial increase in BOD up to 15 days followed by a decrease after 20 days. FT-IR and GC-MS data also reveal that the initial components in the untreated effluent disappear after 20 days of treatment, confirming biodegradation of the dye. Phytotoxicity tests on the untreated effluent samples using...

2006-01-01

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

281

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

282

Role of mitochondria in sulfolipid biosynthesis by Euglena chloroplasts

Response of a non-Hodgkin lymphoma to "6"0Co therapy monitored by 31P MRS in situ

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

283

Exploring the structural dynamics of the E. coli chaperonin GroEL using translation-libration-screw crystallographic refinement of intermediate states

High quality "3"1P MR spectra (signal to noise ratio (S/N) approximately 18, 15 min acquisition for each spectrum) were consistently obtained with surface coils over a period of 6-week RT. Both transient and steady state alterations in metabolites in response to RT were found in this case. The transient changes occurred during the first 3 hr immediately after the 3rd fractionated RT, these changes include the transient elevation of the PCr resonance, a decrease in PDE and an increase in intracellular pH. The monitoring showed that the metabolites approached steady state approximately 2 hr after the fractionated radiation intervention, suggesting that in vivo MRS can be useful for studying the dynamics of tumor response to RT such as repair of potential lethal damage, growth delay, and reoxygenation etc. The steady-state MR spectra showed the net response to each intervention and can clinically be useful for predicting and measuring the result of the fractionated RT. In this case study, ...

284

Salicylate Treatment Improves Age-Associated Vascular Endothelial Dysfunction: Potential Role of Nuclear Factor kB and Forkhead Box O Phosphorylation

Large rigid-body domain movements are critical to GroEL-mediated protein folding, especially apical domain elevation and twist associated with the formation of a folding chamber upon binding ATP and co-chaperonin GroES. Here, we have modeled the anisotropic displacements of GroEL domains from various crystallized states, unliganded GroEL, ATP?S-bound, ADP-AlFx/GroES-bound, and ADP/GroES bound, using translation-libration-screw (TLS) analysis. Remarkably, the TLS results show that the inherent motions of unliganded GroEL, a polypeptide-accepting state, are biased along the transition pathway that leads to the folding-active state. In the ADP-AlFx/GroES-bound folding-active state the dynamic modes of the apical domains become reoriented and coupled to the motions of bound GroES. The ADP/GroES complex exhibits these same motions, but they are increased in magnitude, potentially reflecting the decreased stability of the complex after nucleotide ...

2004-08-12

285

British Library Electronic Table of Contents (United Kingdom)

We hypothesized that I kappa B kinase (IKK)-mediated nuclear factor kappa B and forkhead BoxO3a phosphorylation will be associated with age-related endothelial dysfunction. Endothelium-dependent dilation and aortic protein expression/phosphorylation were determined in young and old male B6D2F1 mice and old mice treated with the IKK inhibitor, salicylate. IKK activation was greater in old mice and was associated with greater nitrotyrosine and cytokines. Endothelium-dependent dilation, nitric oxide (NO), and endothelial NO synthase phosphorylation were lower in old mice. Endothelium-dependent dilation and NO bioavailability were restored by a superoxide dismutase mimetic. Nuclear factor kappa B and forkhead BoxO3a phosphorylation were greater in old and were associated with increased express...

2011-01-01

286

Impairments of astrocytes are involved in the D-galactose-induced brain aging

Gene discovery in the Acanthamoeba castellanii genome

Astrocyte dysfunction is implicated in course of various age-related neurodegenerative diseases. Chronic injection of D-galactose can cause a progressive deterioration in learning and memory capacity and serve as an animal model of aging. To investigate the involvement of astrocytes in this model, oxidative stress biomarkers, biochemical and pathological changes of astrocytes were examined in the hippocampus of the rats with six weeks of D-galactose injection. D-galactose-injected rats displayed impaired antioxidant systems, an increase in nitric oxide levels, and a decrease in reduced glutathione levels. Consistently, western blotting and immunostaining of glial fibrillary acidic protein showed extensive activation of astrocytes. Double-immunofluorescent staining further showed activated astrocytes highly expressed inducible nitric oxide synthase. Electron microscopy demonstrated the degeneration of astrocytes, especially in the aggregated area of synapse and ...

2008-05-16

287

Duration-controlled swimming exercise training induces cardiac hypertrophy in mice.

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 breakdown of cellulose were ...

2005-08-01

288

Antifungal thiopeptide cyclothiazomycin B1 exhibits growth inhibition accompanying morphological changes via binding to fungal cell wall chitin

Exercise training associated with robust conditioning can be useful for the study of molecular mechanisms underlying exercise-induced cardiac hypertrophy. A swimming apparatus is described to control training regimens in terms of duration, load, and frequency of exercise. Mice were submitted to 60- vs 90-min session/day, once vs twice a day, with 2 or 4% of the weight of the mouse or no workload attached to the tail, for 4 vs 6 weeks of exercise training. Blood pressure was unchanged in all groups while resting heart rate decreased in the trained groups (8-18%). Skeletal muscle citrate synthase activity, measured spectrophotometrically, increased (45-58%) only as a result of duration and frequency-controlled exercise training, indicating that endurance conditioning was obtained. In groups which received duration and endurance conditioning, cardiac weight (14-25%) and myocyte dimension (13-20%) increased. The best conditioning protocol to promote physiological ...

2003-11-17

289

British Library Electronic Table of Contents (United Kingdom)

Cyclothiazomycin B1 (CTB1) is an antifungal cyclic thiopeptide isolated from the culture broth of Streptomyces sp. HA 125-40. CTB1 inhibited the growth of several filamentous fungi including plant pathogens along with swelling of hyphae and spores. The antifungal activity of CTB1 was weakened by hyperosmotic conditions, and hyphae treated with CTB1 burst under hypoosmotic conditions, indicating increased cell wall fragility. CTB1-sensitive fungal species contain high levels of cell wall chitin and/or chitosan. Unlike nikkomycin Z, a competitive inhibitor of chitin synthase (CHS), CTB1 did not inhibit CHS activity. Although CTB1 inhibited CHS biosynthesis, the same result was also obtained with a non-specific proteins inhibitor, cycloheximide, which did not reduce cell wall rigidity. These ...

2011-01-01

290

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

Reactive biomolecular divergence in genetically altered yeast cells and isolated mitochondria as measured by biocavity laser spectroscopy : a rapid diagnostic method for studying cellular responses to stress and disease.

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. ...

2001-07-01

291

Ethanol production and a case study of ethanol produced from sweet sorghum stalks via solid state fermentation

We report an analysis of four strains of baker's yeast (Saccharomyces cerevisiae) using biocavity laser spectroscopy. The four strains are grouped in two pairs (wild type and altered), in which one strain differs genetically at a single locus, affecting mitochondrial function. In one pair, the wild-type rho+ and a rho0 strain differ by complete removal of mitochondrial DNA (mtDNA). In the second pair, the wild-type rho+ and a rho- strain differ by knock-out of the nuclear gene encoding Cox4, an essential subunit of cytochrome c oxidase. The biocavity laser is used to measure the biophysical optic parameter Deltalambda, a laser wavelength shift relating to the optical density of cell or mitochondria that uniquely reflects its size and biomolecular composition. As such, Deltalambda is a powerful parameter that rapidly interrogates the biomolecular state of single cells and mitochondria. Wild-type cells and mitochondria produce Gaussian-like distributions ...

2006-12-01

292

3-Methyl-3-deazaadenine, a stable isostere of N3-methyl-adenine, is efficiently bypassed by replication in vivo and by transcription in vitro.

Ethanol has excellent fuel properties, such as high octane, high heat of vaporization and low photochemical reactivity in the atmosphere. It is less volatile than gasoline and there is lower smog formation from evaporative emissions of pure ethanol compared to gasoline. As such, ethanol has emerged as an important alternative energy source that is sustainable, efficient, cost effective, convenient and safe. In 2006, global production of ethanol reached 13.5 billion gallons, up from 12.1 billion gallons in 2005. However, in light of the current debate of food versus fuel, the industry must shift to non-food feedstocks. This paper described an emerging technology to cost-effectively produce ethanol from sweet sorghum stalks, the most promising alternative feedstock to corn, via solid state fermentation (SSF). Experiments of advanced solid state fermentation (ASSF) for ethanol production from sweet sorghum by Saccharomyces cerevisiae were conducted in laboratory and pilot scales studies. ...

2008-07-01

293

T-2 mycotoxin inhibits mitochondrial protein synthesis

The goal of the present work was to determine the impact of N3-methyladenine (3-mA), an important lesion generated by many environmental agents and anticancer drugs, on in vivo DNA replication and in vitro RNA transcription. Due to 3-mA chemical instability, the stable isostere 3-methyl-3-deazaadenine (3-m-c(3)A) was site specifically positioned into an oligodeoxynucleotide. The oligomer was, then incorporated into a vector system that is rapidly converted to ssDNA inside yeast cells and requires DNA replication opposite the lesion for plasmid clonal selection. For control purposes, an adenine or a stable apurinic/apyrimidinic (AP)-lesion was placed at the same site. The presence of each lesion in the oligonucleotide was confirmed by MALDI-TOF analysis. Plasmids were then transfected into yeast cells. While the AP-site dramatically reduced plasmid replication in all strains, the 3-m-c(3)A had a slight effect in the rad30 background which ...

2011-06-14

294

Study on the variations of molecular structures of some biomolecules induced by free electron laser using FTIR spectroscopy

The authors investigated the effect of T-2 toxin on rat liver mitochondrial protein synthesis. Isolated rat liver mitochondria were supplemented with an S-100 supernatant from rat liver and an external ATP-generating system. An in-vitro assay employing cycloheximide, and inhibitor of cytoplasmic protein synthesis, and chloramphenicol, and inhibitor of mitochondrial protein synthesis, to distinguish mitochondrial protein synthesis from the cytoplasmic process. Amino acid incorporation into mitochondria was dependent on the concentration of mitochondria and was inhibited by chloramphenicol. The rate of uptake of tritium leucine into mitochondrial protein was unaffected by the addition of T-2 toxin and was not a rate-limiting step in incorporation. However, 0.02 micrograms/ml of T-2 toxin decreased the rate of protein synthesis inhibition correlated with the amount of T-2 toxin taken up by the mitochondria. While T-2 toxin is known to inhibit eukaryotic protein ...

1988-01-01

295

Protons and glucose metabolism in shock

In this study, free electron laser (FEL) with selective wavelength was used to induce structure changes of biomolecules, which were characterized by FTIR spectroscopy. For understanding of the interactions between FEL and biomolecules as well as biological tissues, the biomolecules investigated are ATP, ADP, AMP, t-RNA, D-ribose and the complex of SmCl_3-D-ribose. Their FTIR spectra before and after irradiation of FEL show molecular structure variations of the samples after irradiation of FEL, especially the rearrangement of their hydrogen bond networks. Along with the various irradiation wavelengths, irradiation time and molecular structures, the changes after irradiation are different for these molecules. In the FTIR spectra after irradiation, the phenomenon that the bands split into several peaks indicates the existence of several structures, conformations and configurations, which may be prompted by multiple photons process induced by FEL. After irradiation, ...

2007-05-01

296

Phosphorylation of proteins in Clostridium thermohydrosulfuricum

When oxygen is limiting, animals can ferment glucose via several metabolic pathways varying in energetic efficiency and leading to various end products (such as lactate, succinate, or propionate). Because the pH dependence of H/sup +/ production by fermentation is opposite to that by hydrolysis of adenosine triphosphate formed in the fermentation, the total number of moles of protons generated is always two per mole of fermentable substrate. However, two and three times more adenosine triphosphate can be turned over per mole of protons produced in succinate and propionate fermentations, respectively, than in lactate fermentation. At its limit, this advantage would achieve the same balance between H/sup +/ production and H/sup +/ consumption during ATP cycling that is observed in aerobic metabolism, a situation observed in certain alcohol fermentations. Since proton balance during anaerobiosis is clearly adaptable, we consider possible impact and functions of net ...

1983-01-01

297

K"A"T"P channels in mesenchymal stromal stem cells: Strong up-regulation of Kir6.2 subunits upon osteogenic differentiation

Cell extracts of the thermophile Clostridium thermohydrosulfuricum catalyzed the phosphorylation by (..gamma..-/sup 32/P)ATP of several endogenous proteins with M/sub r/s between 13,000 and 100,000. Serine and tyrosine were the main acceptors. Distinct substrate proteins were found in the soluble (e.g., proteins p66, p63, and p53 of M/sub r/s 66,000, 63,000, and 53,000, respectively) and particulate (p76 and p30) fractions, both of which contained protein kinase and phosphatase activity. The soluble fraction suppressed the phosphorylation of particulate proteins and contained a protein kinase inhibitor. Phosphorylation of p53 was promoted by 10..mu..M fructose 1,6-bisphosphate or glucose 1,6-bisphosphate and suppressed by hexose monophosphates, whereas p30 and p13 were suppressed by 5 ..mu..M brain (but not spinach) calmodulin. Polyamines, including the odd polyamines characteristic of thermophiles, modulated the labeling of most of the phosphoproteins. Apart from ...

1986-02-01

298

British Library Electronic Table of Contents (United Kingdom)

The promising use of mesenchymal stromal cells (MSC) in regenerative technologies accounts for necessity of detailed study of their physiology. Proliferation and differentiation of multipotent cells often involve changes in their metabolic state. In the present study, we analyzed the expression of ATP-sensitive potassium (K"A"T"P) channels in MSC and upon in vitro differentiation. K"A"T"P channels are present in many cells and regulate a variety of cellular functions by coupling cell metabolism with membrane potential. Kir6.1, Kir6.2 and SUR2A were expressed in undifferentiated MSC, whereas SUR2B and SUR1 were not detected on cDNA and protein level. Upon adipogenic differentiation Kir6.1 and SUR2A showed a significant reduction of the amount of mRNA by 84% and 95%, respectively, whereas Ki...

2011-01-01

299

Intra-cellular transport of single-headed molecular motors KIF1A

CERN Document Server

Motivated by recent experiments on KIF1A, a representative member of single-headed kinesin motor proteins family, we develop a theoretical model of intra-cellular transport by mutually interacting molecular motors. The model explicitly accounts not only for the hydrolysis of ATP, but also for the ratchet mechanism which is believed to drive each individual KIF1A motor. We study the model by a combination of analytical and numerical techniques. A remarkable feature of this model is that all the parameters in it can be completely evaluated from experimental data. Our results in the dilute limit are in excellent quantitative agreement with the empirical data from single molecule experiments. In the high density regime the predictions of the model also agree qualitatively with the corresponding experimental observations. We derive a phase diagram that shows the influence of hydrolysis and Langmuir kinetics on the collective spatio-temporal organization of the motors. ...

2005-01-01

300

Grain boundary self-diffusion of alloy 800 as affected by sulphur, phosphorous and carbon

Grain boundary self-diffusion of alloy 800 as affected by sulphur, phosphorous and carbon

Using a radioactive tracer method the bulk and grain boundary diffusion of {sup 59}Fe was determined in industrial alloy 800 and melts of alloy 800 with additional P and S in the temperature range 800 to 1000 C. The use of the approximation of Suzuoka was confirmed by autoradiographs. In alloy 800 H the activation energy of grain boundary diffusion of {sup 59}Fe is (209 {+-} 17)kJ/mol. Dissolved elements especially P increase the activation energy of the grain boundary diffusion of Fe by their segregation to the grain boundaries. In addition the influence of the grain boundary diffusion on the growth of creep cavities was investigated in the same materials, and the chemical composition of the creep cavities and grain boundaries were analysed by Auger electron spectroscopy (AES). For alloy 800 + 0.088 wt-%P an enrichment of about 14 at-%P was observed at the grain boundaries. The addition of P clearly enhances the creep strength of alloy 800; this can probably be ...

1999-08-01

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301

Comparison of maintenance energy expenditures and growth yields among several rumen bacteria grown on continuous culture.

Using a radioactive tracer method the bulk and grain boundary diffusion of "5"9Fe was determined in industrial alloy 800 and melts of alloy 800 with additional P and S in the temperature range 800 to 1000 C. The use of the approximation of Suzuoka was confirmed by autoradiographs. In alloy 800 H the activation energy of grain boundary diffusion of "5"9Fe is (209 #+-# 17)kJ/mol. Dissolved elements especially P increase the activation energy of the grain boundary diffusion of Fe by their segregation to the grain boundaries. In addition the influence of the grain boundary diffusion on the growth of creep cavities was investigated in the same materials, and the chemical composition of the creep cavities and grain boundaries were analysed by Auger electron spectroscopy (AES). For alloy 800 + 0.088 wt-%P an enrichment of about 14 at-%P was observed at the grain boundaries. The addition of P clearly enhances the creep strength of alloy 800; this can probably be explained ...

1998-07-06

302

Changes in intracellular Ca2+ levels induced by cytokines and P2 agonists differentially modulate proliferation or commitment with macrophage differentiation in murine hematopoietic cells.

Maintenance energy expenditures were mesured for five rumen bacteria, Selenomonas ruminantium, Butyrivibrio fibrisolvens, Bacteroides ruminicola, Megasphaera elsdenii, and Streptococcus bovis, by using a complex medium with glucose as the carbon source. Large differences (as high as 8.5-fold) in maintenance energy expenditures were seen among these bacteria. The suggestion is made that maintenance requirements could be a significant determinant of bacterial competition in the rumen. Theoretical maximum growth yields, calculated from double reciprocal plots of yield versus dilution rate, were compared to theoretical Y(ATP) values in order to estimate minimum molar adenosine 5'-triphosphate yields from glucose for each bacterium. Results showed that relative yield among the bacteria was growth rate dependent. At high dilution rates, both S. ruminantium and S. bovis produced lactate as their principal fermentation product. At lower dilution rates very little lactate ...

1979-03-01

303

Activation of fat cell adenylate cyclase by protein kinase C

The role of intracellular Ca2+ (Ca2+i) on hematopoiesis was investigated in long term bone marrow cultures using cytokines and agonists of P2 receptors. Cytokines interleukin 3 and granulocyte/macrophage colony stimulator factor promoted a modest increase in Ca2+i concentration ([Ca2+]i) with activation of phospholipase Cgamma, MEK1/2, and Ca2+/calmodulin kinase II. Involvement of protein kinase C was restricted to stimulation with interleukin 3. In addition, these cytokines promoted proliferation (20 times) and an increase in the Gr-1(-)Mac-1+ population with participation of gap junctions (GJ). Nevertheless ATP, ADP, and UTP promoted a large increase in [Ca2+]i, moderate proliferation (6 times), a reduction in the primitive Gr-1(-)Mac-1(-)c-Kit+ population, and differentiation into macrophages without participation of GJ. It is likely that Ca2+i participates as a regulator of hematopoietic signaling: moderate increases in [Ca2+]i would be related to ...

2008-09-05

304

The genetic basis of salinity tolerance in Arctic charr (Salvelinus alpinus).

Purified protein kinase C (C-kinase) from guinea pig pancreas and rat brain stimulated adenylate cyclase activity in purified rat adipocyte membranes. Cyclase stimulation occurred over 100 to 1000 mU/ml of C-kinase activity, required greater than 10 ..mu..M calcium, proceeded without a lag, was not readily reversible, and required no exogenous phospholipid. Moreover, C-kinase inhibitors, such as chlorpromazine and palmitoyl carnitine, inhibited selectively adenylate cyclase which was activated by C-kinase and calcium. Depending on assay conditions, 10 nM 12-0-tetradecanoylphorbol-13-acetate (TPA) either enhanced or was required for kinase action on cyclase. Also, TPA plus calcium promoted the quantitative association of C-kinase with membranes. Adenylate cyclase activation by C-kinase was seen both in the presence and absence of exogenous GTP, indicating that the kinase effect does not result from an action on the GTP-binding, inhibitory regulatory component (N/sub i/) of the cyclase ...

1986-05-01

305

Studies on the biosorption of uranium by a thermotolerant, ethanol-producing strain of Kluyveromyces marxianus

ABSTRACT: BACKGROUND: The capacity to maintain internal ion homeostasis amidst changing conditions is particularly important for teleost fishes whose reproductive cycle is dependent upon movement from freshwater to seawater. Although the physiology of seawater osmoregulation in mitochondria-rich cells of fish gill epithelium is well understood, less is known about the underlying causes of inter- and intraspecific variation in salinity tolerance. We used a genome-scan approach in Arctic charr (Salvelinus alpinus) to map quantitative trait loci (QTL) correlated with variation in four salinity tolerance performance traits and six body size traits. Comparative genomics approaches allowed us to infer whether allelic variation at candidate gene loci (e.g., ATP1alpha1b, NKCC1, CFTR, and cldn10e) could have underlain observed variation. RESULTS: Combined parental analyses yielded genome-wide significant QTL on linkage groups 8, 14 and 20 for salinity tolerance performance ...

2011-09-21

306

Potential anti fungal agents. Synthesis and activity of 2-alkylthiopyridine-4-carbothioamides

The ability of residual biomass from the thermotolerant ethanol-producing yeast strain Kluyveromyces marxianus IMB3 to function as a biosorbent for uranium has been examined. It was found that the biomass had an observed maximum biosorption capacity of 120 mg U/g dry weight of biomass. The calculated value for the biosorption maximum, obtained by fitting the data to the Langmuir model was found to be 130 mg U/g dry weight biomass. Maximum biosorption capacities were examined at a number of temperatures and both the observed and calculated values obtained for those capacities increased with increasing temperature. Decreasing the pH of the biosorbate solution resulted in a decrease in uptake capacity. When biosorption reactions were carried out using sea-water as the diluent it was found that the maximum biosorption capacity of the biomass increased significantly. Using transmission electron microscopy, uranium crystals were shown to be concentrated on the outer ...

1997-06-01

307

NADP Regulates the Yeast GAL Induction System

A series of 2-alkylthiopyrine-4-carbothioamides were synthesized, and their anti-fungal potency was tested. The chemical structures were proved by infrared spectroscopy (IR) and {sup 1}H nuclear magnetic resonance ({sup 1}H-NMR) data and by elemental analysis. The minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) assessment were used for the estimation of potential activity in vitro. The study comprising 21 clinical isolates of fungi showed that two compounds exhibited fair inhibitory activity against some yeasts and dermatophytes. Selective fungistatic activity against non-dermatophytes (MIC = 3.12-25.0 {mu}g/mL) was found also in another compound. None of the above compounds showed inhibitory activity against non-dermatophyte filamentous fungi. Microbiological activity of 2-alkylthiopyridine-4-carbothioamides appears to be mainly related to hydrophobicity of alkyl in position 2. (authors). 10 refs., 8 tabs.

1996-05-01

308

Molecular studies of the uncoupling protein

Transcriptional regulation of the galactose-metabolizing genes in Saccharomyces cerevisiae depends on three core proteins: Gal4p, the transcriptional activator that binds to upstream activating DNA sequences (UASGAL); Gal80p, a repressor that binds to the carboxyl terminus of Gal4p and inhibits transcription; and Gal3p, a cytoplasmic transducer that, upon binding galactose and adenosine 5'-triphosphate, relieves Gal80p repression. The current model of induction relies on Gal3p sequestering Gal80p in the cytoplasm. However, the rapid induction of this system implies that there is a missing factor. Our structure of Gal80p in complex with a peptide from the carboxyl-terminal activation domain of Gal4p reveals the existence of a dinucleotide that mediates the interaction between the two. Biochemical and in vivo experiments suggests that nicotinamide adenine dinucleotide phosphate (NADP) plays a key role in the initial induction event.

2008-01-01

309

Evaluation of heavy-metal ion toxicity in fish cells using a combined stress protein and cytotoxicity assay

The uncoupling protein (UCP) is a proton/anion transporter found in the inner mitochondrial membrane of brown adipocyte. Although UCP has nor been detected in mitochondria from any other tissue, it shares structural and catalytic properties with several other mitochondrial carrier proteins. Although UCP was discovered only recently it is one of the most extensively studied mitochondrial carrier proteins.More recently, the mouse, rat, and human genes encoding for UCP have been isolated and sequenced. The availability of these various tools has led to several significant observations. UCP gene expression is strongly controlled at the level of transcription by signals that are activated after the stimulation of brown adipocytes by norepinephrine. The comparison of UCP gene with the genes encoding the adenine nucleotide translocator revealed the existence of structural and evolutionary homologies. Moreover, in humans the UCP gene and one form of adenine nucleotide translocator gene are ...

1991-06-01

310

Effects of paraquat on Escherichia coli: Differences between B and K-12 strains

All organisms, from bacteria and yeast to humans, respond to physical and chemical stressors by increasing the synthesis of a small group of cellular stress proteins.'' The authors have developed a simple in vitro system for quickly screening environmentally relevant stressors to detect stress-induced proteins that are good candidates for biomarkers. Polyacrylamide gel electrophoresis was used to detect stressor-induced, concentration-dependent changes in cellular stress protein levels in two fish cell culture systems, whereas simultaneous in vitro neutral red uptake cytotoxicity assays measured the stressors effect on cellular physiology. There was a direct concentration-dependent relationship between sublethal cytotoxic effects and the increases in stress protein levels. Increases of 50 to 200% were detected in stress proteins from desert topminnow, Poeciliopsis lucida, hepatoma-derived cell cultures exposed to cadmium or copper. Three proteins ...

1994-08-01

311

Construction of a human MluI YAC library

Escherichia coli B and K-12 are equally susceptible to the bacteriostatic effects of aerobic paraquat, but they differed strikingly when the lethality of paraquat was evaluated. E. coli B suffered an apparent loss of viability when briefly exposed to paraquat, whereas E. coli K-12 did not. This difference depended on the ability of the B-strain, but not the K-12 strain, to retain internalized paraquat; the B strain was killed on aerobic tryptic soy-yeast extract plates during the incubation which preceded the counting of colonies. This difference in retention of paraquat between strains was demonstrated by delayed loss of viability, by growth inhibition, and by cyanide-resistant respiration after brief exposure to paraquat, washing, and testing in fresh medium. This difference was also shown by using ({sup 14}C)paraquat. This previously unrecognized difference between E. coli B and K-12 has been the cause of apparently contradictory reports and should lead to some ...

1990-02-01

312

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

313

Biosorption of Ni (II) by Schizosaccharomyces pombe: kinetic and thermodynamic studies

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

314

British Library Electronic Table of Contents (United Kingdom)

The potential of the dried yeast, wild-type Schizosaccharomyces pombe, to remove Ni(II) ion was investigated in batch mode under varying experimental conditions including pH, temperature, initial metal ion concentration and biosorbent dose. Optimum pH for biosorption was determined as 5.0. The highest equilibrium uptake of Ni(II) on S. pombe, q e, was obtained at 25??C as 33.8?mg?g?1. It decreased with increasing temperature within a range of 25?50??C denoting an exothermic behaviour. Increasing initial Ni(II) concentration up to 400?mg?L?1 also elevated equilibrium uptake. No more adsorption took place beyond 400?mg?L?1. Equilibrium data fitted better to Langmuir model rather than Freundlich model. Sips, Redlich?Peterson, and Kahn isotherm equations modelled the investigated system with a...

2011-01-01

315

The effect of elevated carbon dioxide on a Sierra-Nevadan dominant species: Pinus ponderosa

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

The impact of increasing atmospheric C0{sub 2} has not been fully evaluated on western coniferous forest species. Two year old seedlings of Pinusponderosa were grown in environmentally controlled chambers under increased C0{sub 2} conditions for 6 months. These trees exhibit morphological, physiological, and biochemical alterations when compared to our controls. Analysis of whole plant biomass distribution has shown no significant effect to the root to shoot ratios, however needles subjected to elevated C0{sub 2} exhibited an increased overall specific needle mass and a decreased total needle area. Morphological changes at the needle level included decreased mesophyll to vascular tissue 91 ratio and variations in starch storage in chloroplasts. The elevated CO{sub 2} increased internal CO{sub 2} concentrations and assimilation of carbon. Biochemical assays revealed that ribulose-bisphosphate carboxylase specific activities increased on per unit area basis with C0{sub 2} treatment ...

1995-01-01

316

Regulation of sucrose synthesis in water stressed leaves

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

317

Gene expression profiles deciphering rice phenotypic variation between Nipponbare (Japonica) and 93-11 (Indica) during oxidative stress.

Alteration in carbon metabolism and carbohydrate partitioning occur in drought stressed plants. Some species accumulate large quantities of starch in the chloroplast, which may be used to support sucrose synthesis under conditions of limited carbon supply. The authors monitored chemical partitioning of carbon between sugars and starch and the activity of sucrose phosphate synthase (SPS) and fructose 1,6 bisphosphatase (FBPase) in the source leaves of water stressed tomatoes. Plants were stressed by withdrawing water for 10 days and rewatered for recovery. Water potential dropped from {minus}0.8 to {minus}2.2MPA in 10 days, but recovered to control level 2 days after rewatering. Photosynthetic rates as measured by the activity of Rubisco followed similar patterns to those of water potential. After 10 days, leaf starch levels decreased to less than 50% of control. Sucrose levels did not increase significantly, but hexose levels increased 3-4 fold during the stress ...

1991-05-01

318

Antioxidant treatment with quercetin ameliorates erectile dysfunction in streptozotocin-induced diabetic rats.

Rice is a very important food staple that feeds more than half the world's population. Two major Asian cultivated rice (Oryza sativa L.) subspecies, japonica and indica, show significant phenotypic variation in their stress responses. However, the molecular mechanisms underlying this phenotypic variation are still largely unknown. A common link among different stresses is that they produce an oxidative burst and result in an increase of reactive oxygen species (ROS). In this study, methyl viologen (MV) as a ROS agent was applied to investigate the rice oxidative stress response. We observed that 93-11 (indica) seedlings exhibited leaf senescence with severe lesions under MV treatment compared to Nipponbare (japonica). Whole-genome microarray experiments were conducted, and 1,062 probe sets were identified with gene expression level polymorphisms between the two rice cultivars in addition to differential expression under MV treatment, which were assigned as Core Intersectional Probesets ...

2010-01-08

319

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

Oxidative stress is demonstrated to be involved in the pathophysiological mechanism of erectile dysfunction (ED). Quercetin, a potent bioflavonoid, has been reported to have the antioxidant role. In the present study, we examined the effect of quercetin on ED and oxidative stress in streptozotocin (STZ)-induced diabetic rats. Diabetes was induced in Sprague-Dawley rats with a single intravenous injection of STZ. The diabetic rats were then randomized to diabetic group and quercetin therapy groups which were treated with quercetin at different doses of 5, 20 and 50mg/kg per day respectively. At the end of the 8th week, erectile function was assessed by measuring the rise in intracavernous pressure (ICP) following cavernous nerve electrostimulation. Superoxide dismutase (SOD) activity, thiobarbituric acid-reacting substance (TBARS) and nitrite and nitrate (NOx) levels were measured in cavernosum tissue. Endothelial NO synthase (eNOS) expression was determined using ...

2011-06-14

320

Small-molecule screen identifies inhibitors of a human intestinal calcium-activated chloride channel.

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 activity of pyruvate ...

2005-03-01

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321

Response of photosynthesis, starch and sucrose synthesis to increased CO sub 2 and lowered O sub 2

Calcium-activated chloride channels (CaCCs) are widely expressed in mammalian tissues, including intestinal epithelia, where they facilitate fluid secretion. Potent, selective CaCC inhibitors have not been available. We established a high-throughput screen for identification of inhibitors of a human intestinal CaCC based on inhibition of ATP/carbachol-stimulated iodide influx in HT-29 cells after lentiviral infection with the yellow fluorescent halide-sensing protein YFP-H148Q/I152L. Screening of 50,000 diverse, drug-like compounds yielded six classes of putative CaCC inhibitors, two of which, 3-acyl-2-aminothiophenes and 5-aryl-2-aminothiazoles, inhibited by >95% iodide influx in HT-29 cells in response to multiple calcium-elevating agonists, including thapsigargin, without inhibition of calcium elevation, calcium-calmodulin kinase II activation, or cystic fibrosis transmembrane conductance regulator chloride channels. These compounds also inhibited ...

2007-12-14

322

Open test assembly (OTA) shear demonstration testing work/test plan

Increasing photosynthesis rate by raising CO{sub 2} and lowering O{sub 2} usually results in measured rates lower than those expected from the kinetics of rubisco. The failure of leaves to utilize the increased efficiency of rubisco has been attributed to limitations in rates of either starch or sucrose synthesis, which lead to a lowering of Pi and a reduction in the rate of ATP synthesis. In sugar beet leaves, increasing CO{sub 2} from 350 to 600 {mu}L L{sup {minus}1} usually increased photosynthesis only about 10%, compared to the 45% expected, and lowering O{sub 2} from 2 to 21% increased photosynthesis only an additional 20%, compared to the additional 40% expected. In some cases, raising CO{sub 2} increased photosynthesis the full 30% with no further increase occurring upon lowering O{sub 2}. In all cases, rates of starch and sucrose synthesis consistently increased in proportion to increasing photosynthesis rate. Metabolites that are predominantly cytosolic ...

1990-05-01

323

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

This document describes the development testing phase associated with the OTA Shear activity and defines the controls to be in place throughout the testing. The purpose of the OTA Shear Program was to provide equipment that is needed for the processing of 40 foot long, sodium wetted, irradiated core components previously used in the FFTF reactor to monitor fuel and materials tests. There are currently 15 of these OTA test stalks located in the Test Assembly Conditioning Station (TACS) inerted vault. These need to be dispositioned for a shutdown mission to eliminate this highly activated, high dose inventory prior to turnover to the ERC since they must be handled by remote operations. These would also need to be dispositioned for a restart mission to free up the vault they currently reside in. The waste handling and cleaning equipment in the J33M Cell was designed and built for the handling of reactor components up to the standard 12 foot length. This program will provide the equipment ...

1998-07-16

324

Internalized insulin-receptor complexes are unidirectionally translocated to chloroquine-sensitive degradative sites. Dependence on metabolic energy

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 acid (mABS) linked on the triazine ...

2007-01-01

325

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

Insulin receptors on the surface of isolated rat adipocytes were photoaffinity labeled at 12 degrees C with the iodinated photoreactive insulin analogue, 125I-B2 (2-nitro-4-azidophenylacetyl)-des-PheB1-insulin, and the pathways in the intracellular processing of the labeled receptors were studied at 37 degrees C. During 37 degrees C incubations, the labeled 440-kDa insulin receptors were continuously internalized (as assessed by trypsin inaccessibility) and degraded such that up to 50% of the initially labeled receptors were lost by 120 min. Metabolic poisons (0.125-0.75 mM 2,4-dinitrophenol (DNP) and 1-10 mM NaF), which led to dose-dependent depletion of adipocyte ATP pools, inhibited receptor loss, and caused up to 3-fold increase in intracellular receptor accumulation. This effect was due to inhibition of intracellular receptor degradation, and there was no apparent effect of the metabolic poisons on initial internalization of the receptors. Following maximal ...

1988-04-25

326

Direct photoaffinity labeling of gizzard myosin with ["3H]uridine diphosphate places Glu185 of the heavy chain at the active site

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 did not differ between ...

1989-04-01

327

Digitonin abolishes free 2-deoxy-D-glucose accumulation in isolated rat adipocytes

The active site of chicken gizzard myosin was labeled by direct photoaffinity labeling with ["3H]UDP. ["3H] UDP was stably trapped at the active site by addition of vanadate (Vi) and Co"2"+. The extraordinary stability of the myosin.Co2+.[3H]UDP.Vi complex (t1/2 greater than 5 days at 0 degrees C) allowed it to be purified free of extraneous ["3H]UDP before irradiation began. Upon UV irradiation, greater than 60% of the trapped ["3H]UDP was photoincorporated into the active site. Only the 200-kDa heavy chain was labeled, confirming earlier results using ["3H]UTP. Extensive tryptic digestion of photolabeled myosin subfragment 1 followed by high performance liquid chromatography separations and removal of nucleotide phosphates by treatment with alkaline phosphatase allowed two labeled peptides to be isolated. Sequencing of the labeled peptides and radioactive counting showed that Glu185 was the residue labeled. Since UDP is a zero-length cross-linker, Glu185 is located at the ...

328

Chelation of intracellular calcium blocks insulin action in the adipocyte

The hypothesis that accumulation against sizable chemical gradients of free (non-phosphorylated) 2-deoxy-D-glucose (2dGlc) in isolated rat adipocytes results from an intracellular compartmentation of free hexose was investigated. Cells exposed to 20 ..mu..g/ml digitonin for 10' demonstrated an increased plasma membrane permeability indexed by increased L-glucose entry rates and cellular (presumably cytosolic) protein and K/sup +/ loss. Functional integrity of intracellular organelles was indicated by the ability of the cells to support ATP-driven /sup 45/Ca/sup 2 +/-uptake. Equilibrium 3-O-methylglucose (3-O-MG, a non-accumulated hexose) levels were unaffected. These data suggest a specific permeabilizing action of digitonin at the plasma membrane having no effect on intracellular organelles or passively distributed solutes. Upon addition of digitonin, free 2dGlc fell from 66.5 +/- 8.9 to 7.4 +/- 2.3 pmol/10/sup 5/ cells, a value not significantly ...

1986-03-05

329

Chelation of intracellular calcium blocks insulin action in the adipocyte

The hypothesis that intracellular Ca/sup 2 +/ is an essential component of the intracellular mechanism of insulin action in the adipocyte was evaluated. Cells were loaded with the Ca/sup 2 +/ chelator quin-2, by preincubating them with quin-2 AM, the tetrakis(acetoxymethyl) ester of quin-2. Quin-2 loading inhibited insulin-stimulated glucose transport without affecting basal activity. The ability of insulin to stimulate glucose uptake in quin-2-loaded cells could be partially restored by preincubating cells with buffer supplemented with 1.2 mM CaCl/sub 2/ and the Ca/sup 2 +/ ionophore A23187. These conditions had no effect on basal activity and omission of CaCl/sub 2/ from the buffer prevented the restoration of insulin-stimulated glucose uptake by A23187. Quin-2 loading also inhibited insulin-stimulated glucose oxidation and the ability of insulin to inhibit cAMP-stimulated lipolysis without affecting their basal activities. Incubation of cells with 100 ..mu..M quin-2 or quin-2 AM had ...

1987-02-01

330

Altered protein phosphorylation in sciatic nerve from rats with streptozocin-induced diabetes

The hypothesis that intracellular Ca"2"+ is an essential component of the intracellular mechanism of insulin action in the adipocyte was evaluated. Cells were loaded with the Ca"2"+ chelator quin-2, by preincubating them with quin-2 AM, the tetrakis(acetoxymethyl) ester of quin-2. Quin-2 loading inhibited insulin-stimulated glucose transport without affecting basal activity. The ability of insulin to stimulate glucose uptake in quin-2-loaded cells could be partially restored by preincubating cells with buffer supplemented with 1.2 mM CaCl_2 and the Ca"2"+ ionophore A23187. These conditions had no effect on basal activity and omission of CaCl_2 from the buffer prevented the restoration of insulin-stimulated glucose uptake by A23187. Quin-2 loading also inhibited insulin-stimulated glucose oxidation and the ability of insulin to inhibit cAMP-stimulated lipolysis without affecting their basal activities. Incubation of cells with 100 #mu#M quin-2 or quin-2 AM had no effect on intracellular ...

331

ATRX ADD domain links an atypical histone methylation recognition mechanism to human mental-retardation syndrome

The effect of experimental diabetes on the phosphorylation of proteins in the rat sciatic nerve was studied. Nerves from animals made diabetic with streptozocin were incubated in vitro with (/sup 32/P)orthophosphate and divided into segments from the proximal to the distal end, and proteins from each segment were then separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The principal labeled species were the major myelin proteins, P0, and the basic proteins. After 6 wk of diabetes, the incorporation of isotope into these proteins rose as a function of distance along the nerve in a proximal to distal direction and was significantly higher at the distal end compared with incorporation into nerves from age-matched controls. The overall level of isotope uptake was similar in nerves from diabetic animals and weight-matched controls. The distribution of /sup 32/P among proteins also differed in diabetic nerve compared with both control groups in that P0 and the small basic ...

1987-11-01

332

Neurotoxicity of low-dose repeatedly intranasal instillation of nano- and submicron-sized ferric oxide particles in mice

ATR-X (alpha-thalassemia/mental retardation, X-linked) syndrome is a human congenital disorder that causes severe intellectual disabilities. Mutations in the ATRX gene, which encodes an ATP-dependent chromatin-remodeler, are responsible for the syndrome. Approximately 50% of the missense mutations in affected persons are clustered in a cysteine-rich domain termed ADD (ATRX-DNMT3-DNMT3L, ADD{sub ATRX}), whose function has remained elusive. Here we identify ADD{sub ATRX} as a previously unknown histone H3-binding module, whose binding is promoted by lysine 9 trimethylation (H3K9me3) but inhibited by lysine 4 trimethylation (H3K4me3). The cocrystal structure of ADD{sub ATRX} bound to H3{sub 1-15}K9me3 peptide reveals an atypical composite H3K9me3-binding pocket, which is distinct from the conventional trimethyllysine-binding aromatic cage. Notably, H3K9me3-pocket mutants and ATR-X syndrome mutants are defective in both H3K9me3 binding and localization at ...

2011-07-19

333

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

Olfactory tract has been demonstrated to be an important portal for inhaled solid nanoparticle transportation into the central nervous system (CNS). We have previously demonstrated that intranasally instilled Fe_2O_3 nanoparticles could transport into the CNS via olfactory pathway. In this study, we investigated the neurotoxicity and size effect of repeatedly low-dose (130 #mu#g) intranasal exposure of nano- and submicron-sized Fe_2O_3 particles (21 nm and 280 nm) to mice. The biomarkers of oxidative stress, activity of nitric oxide synthases and release of monoamine neurotransmitter in the brain were studied. Our results showed that significant oxidative stress was induced by the two sizes of Fe_2O_3 particles. The activities of GSH-Px, Cu,Zn-SOD, and cNOS significantly elevated and the total GSH and GSH/GSSG ratio significantly decreased in the olfactory bulb and hippocampus after the nano- and submicron-sized Fe_2O_3 particle treatment (p < 0.05). The ...

2009-01-01

334

The effect of pulse voltage and capacitance on biosorption of uranium by biomass derived from whiskey distillery spent wash

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 conversion of chorismate to prephenate without the accumulation of any enzyme-free intermediates. The kinetic data of the wild-type chorismate mutase ...

335

Quality changes of irradiated spices stored in different indigenous packaging materials

Biosorption of uranium by residual biomass from The Old Bushmill`s Distillery Co. Ltd., Bushmills, Co. Antrim, Northern Ireland, following exposure to short and intense electric pulses has been examined. The biomass was prepared from the distillery spent wash and consisted of non-viable yeast and bacterial cells. As shown previously, untreated biomass had a maximum biosorption capacity of 170 mg uranium/g dry weight biomass. When biosorption reactions were placed between two electrodes and exposed to electric pulses with field strengths ranging from 1.25-3.25 kV/cm at a capacitance of 25 {mu}F, biosorption increased from 170 mg of uranium to 275 mg uranium/g dry weight biomass. The data were obtained from biosorption isotherm analyses and taken as the degree of biosorption at residual uranium concentrations of 3 mM. In addition, when the capacitance of the electric pulses increased from 0.25 {mu}F to 25 {mu}F at a fixed pulse field strength the degree of ...

1999-01-01

336

Multicolor FISH mapping with Alu-PCR-amplified YAC clone DNA determines the order of markers in the BRCA1 region on chromosome 17q12-q21

The changes in quality of irradiated spices, i.e., nutmeg (Myristica fragrans), black pepper and white pepper (Piper nigrum) stored in different indigenous packaging materials were evaluated. Packaging materials used were tin cans; tin can lined with polypropylene film, tin can lined with wrapping paper, woven polypropylene bag and woven polypropylene bag lined with polypropylene film. Packaging in woven polypropylene bag could cause significant changes in a_w-values and moisture contents of the spices. In the tin can, no significant change was found during storage. By applying a polypropylene film liner inside the plastic bag, the changes could be retarded. The other parameters observed, i.e. total plate count, total mould and yeast count and organoleptic scores of the samples were not significantly affected by the packaging method. A dose of 5 kGy could reduce the microbial load of the spices by about 2-3 log cycles. A significant decrease in a_w-value and ...

1990-03-01

337

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

A gene designated BRCA1, implicated in the susceptibility to early-onset familial breast cancer, has recently been localized to chromosome 17q12-q21. To date, the order of DNA markers mapped within this region has been based on genetic linkage analysis. The authors report the use of multicolor fluorescence in situ hybridization to establish a physically based map of five polymorphic DNA markers and 10 cloned genes spanning this region. Three cosmid clones and Alu-PCR-Generated products derived from 12 yeast artificial chromosome clones representing each of these markers were used in two-color mapping experiments to determine an initial proximity of markers relative to each other on metaphase chromosomes. Interphase mapping was then employed to determine the order and orientation of closely spaced loci by direct visualization of fluorescent signals following hybridization of three probes, each detected in a different color. Statistical analysis of the combined data ...

1993-09-01

338

Functions of mammalian Cdc7 kinase in initiation/monitoring of DNA replication and development

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

339

Caldicellulosiruptor obsidiansis sp. nov., an anaerobic, extremely thermophilic, cellulolytic bacterium isolated from Obsidian Pool, Yellowstone National Park

Cdc7 kinase plays an essential role in firing of replication origins by phosphorylating components of the replication complexes. Cdc7 kinase has also been implicated in S phase checkpoint signaling downstream of the ATR and Chk1 kinases. Inactivation of Cdc7 in yeast results in arrest of cell growth with 1C DNA content after completion of the ongoing DNA replication. In contrast, conditional inactivation of Cdc7 in undifferentiated mouse embryonic stem (ES) cells leads to growth arrest with rapid cessation of DNA synthesis, suggesting requirement of Cdc7 functions for continuation of ongoing DNA synthesis. Furthermore, loss of Cdc7 function induces recombinational repair (nuclear Rad51 foci) and G2/M checkpoint responses (inhibition of Cdc2 kinase). Eventually, p53 becomes highly activated and the cells undergo massive p53-dependent apoptosis. Thus, defective origin activation in mammalian cells can generate DNA replication checkpoint signals. Efficient removal of ...

2003-11-27

340

Biological assessments of a mixture of endocrine disruptors at environmentally relevant concentrations in water following UV/H{sub 2}O{sub 2} oxidation

A novel, obligately anaerobic, extremely thermophilic, cellulolytic bacterium, designated OB47T, was isolated from Obsidian Pool, Yellowstone National Park, WY, USA. The isolate was a non-motile, non-spore forming, Gram-positive rod approximately 2 m long by 0.2 m wide and grew at temperatures between 55-85oC with the optimum at 78oC. The pH range for growth was 6.0-8.0 with values of near 7.0 being optimal. Growth on cellobiose produced the fastest specific growth rates at 0.75 hr-1. The organism also displayed fermentative growth on glucose, maltose, arabinose, fructose, starch, lactose, mannose, sucrose, galactose, xylose, arabinogalactan, Avicel, xylan, filter paper, processed cardboard, pectin, dilute acid-pretreated switchgrass and Populus. OB47T was unable to grow on mannitol, fucose, lignin, Gelrite, acetate, glycerol, ribose, sorbital, carboxymethylcellulose and casein. Yeast extract stimulated growth and thiosulfate, sulfate, nitrate, and sulfur were not ...

2010-02-01

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341

Two and 8-azido photoaffinity probes. 1. Enzymatic synthesis, characterization, and biological properties of 2- and 8-azido photoprobes of 2-5A and photolabeling of 2-5A binding proteins

Numerous studies have investigated degradation of individual endocrine disrupting compounds (EDCs) in lab or natural waters. However, natural variations in water matrices and mixtures of EDCs in the environment may confound analysis of the treatment efficiency. Because chemical based analytical methods cannot represent the combined or synergistic activities between water quality parameters and/or the EDC mixtures at environmentally relevant concentrations ({mu}g L{sup -1}-ng L{sup -1}), bioanalytical assessments of residual estrogenic activity in treated water were used to evaluate the performance of the UV based advanced oxidation process for estrogenic contaminants in water. Four EDCs including estradiol (E{sub 2}), ethinyl estradiol (EE{sub 2}), bisphenol-A (BPA) and nonylphenol (NP) were spiked individually or as a mixture at {mu}g L{sup -1}-ng L{sup -1} in laboratory or natural river water. The removal rates of estrogenic activity were quantitatively evaluated by in vitro ...

2007-04-15

342