International Nuclear Information System (INIS)

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

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The marine environment is becoming increasingly contaminated by environmental pollutants with the potential to damage DNA, with marine sediments acting as a sink for many of these contaminants. Understanding genotoxic responses in sediment-dwelling marine organisms, such as polychaetes, is therefore of increasing importance. This study is an exploration of species-specific and cell-specific differences in cell sensitivities to DNA-damaging agents in polychaete worms, aimed at increasing fundamental knowledge of their responses to genotoxic damage. The sensitivities of coelomocytes from three polychaetes species of high ecological relevance, i.e. the lugworm Arenicola marina, the harbour ragworm Nereis diversicolor and the king ragworm Nereis virens to genotoxic damage are compared, and dif...

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International Nuclear Information System (INIS)

Previous experiments had indicated a slight genotoxic potential both in rat and in human colon cells of a sample of 2-dodecylcyclobutanone, a compound formed by irradiation of food containing palmitic acid in its triglycerides. Up to date, there is no evidence that 2-alkylcyclobutanones occur in non-irradiated foodstuffs, consequently it is prudent to test several members of the class of 2-alkylcyclobutanones which are produced by treatment of fat-containing food with ionising radiation. In this work, 2-tetradecylcyclobutanone (derived from stearic acid) has been tested for its cytotoxic and genotoxic potential. Human colon tumor cell lines, i.e. HT 29 and HT 29 clone 19A, were employed as models for in vitro experiments for cytotoxicity and genotoxicity tests. Cytotoxicity was measured by tetrazolium salt reduction assays (MTT and WST-1) and genotoxicity by determining DNA damage using the Comet Assay. ...

UK PubMed Central (United Kingdom)

TCDD and other polyhalogenated aromatic hydrocarbon ligands of the aryl hydrocarbon receptor (AHR) have been classically considered as non-genotoxic compounds because they fail to be directly mutagenic...Full Text Available

UK PubMed Central (United Kingdom)

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N′-nitrosonornicotine (NNN) are tobacco-specific nitrosamines present in tobacco products and smoke. Both compounds...Full Text Available

KoreaScience (Korea)

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Energy Technology Data Exchange (ETDEWEB)

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

Energy Technology Data Exchange (ETDEWEB)

No abstract prepared.

UK PubMed Central (United Kingdom)

Human serum dopamine-β-hydroxylase (3,4-dihydroxyphenylethylamine, ascorbate:oxygen oxidoreductase, EC 1.14.17.1) has been measured in a normal adult population by both enzymatic assay and by...Full Text Available

International Nuclear Information System (INIS)

In patients subjected to bone marrow transplantation the aggressive measures necessarily preceding this treatment lead to drastic metabolic changes which, in turn, are a cause of marked disturbances of homeostasis. The general preparatory measures in the form of cytoreduction by cytostatic drugs are broadened for leukaemic patients to include wholebody irradiation. During the study described here investigations were carried out into the resulting strain on the metabolism. One of the most essential changes observed in those patients was a nitrogen loss of up to approx. 20 g/day that occurred during the immunosuppressive phase. This was accompanied by markedly pronounced variations in fluid elimination, which partly were of an iatrogenic nature. An immediate and sufficient substitution of amino acids in connection with complete parenteral nutrition rapidly acted to compensate for the nitrogen losses, even though these could not be totally reversed to achieve a positive nitrogen balance. ...

British Library Electronic Table of Contents (United Kingdom)

Abstract Aim:- To describe the first case of ceftriaxone-related haemolysis in a patient with congenital nephrotic syndrome (CNS). Background:- Severe haemolysis caused by an immune reaction to ceftriaxone has mostly been described in patients with underlying haematological or immune dysfunction. Case report:- The authors present a 20-month-old boy with CNS of the Finnish type with several previous severe infections treated with ceftriaxone, admitted for suspected sepsis. Following ceftriaxone administration he developed shock secondary to an acute haemolytic reaction, with severe anaemia. Hypersensitivity to ceftriaxone was documented through positive agglutination tests. Conclusion:- Onset of haemolysis following ceftriaxone administration, particularly in a patient previously exposed to...

British Library Electronic Table of Contents (United Kingdom)

Magnesium fructose-1,6-diphosphate is a novel agent of antimyocardial ischaemia. In the present study, the subchronic toxicity of magnesium fructose-1,6-diphosphate was investigated after 13-week repeated intravenous administration in beagle dogs. The animals received doses of 0, 75, 150 and 300 mg/kg/day (three males and three females for each dose). During the study period, clinical signs, mortality, body weights, food consumption, electrocardiogram, urinalysis, haematology, clinical biochemistry, macroscopic findings, organ weights and histopathology were examined. The administration of magnesium fructose-1,6-diphosphate resulted in increased incidence of clinical signs, including salivation and emesis. These effects were transient and were noted in almost all dogs given 300 mg/kg/day a...

UK PubMed Central (United Kingdom)

BackgroundThe molecular mechanisms of variations in individual longevity are not well understood, even though longevity can be increased substantially by means of diverse experimental...Full Text Available

UK PubMed Central (United Kingdom)

Some strains of Bradyrhizobium japonicum have the ability to catabolize indole-3-acetic acid (IAA). Examination of this catabolism in strain 110 by in vivo experiments has revealed an enzymatic activity...Full Text Available

UK PubMed Central (United Kingdom)

Combined QM(PM3)/MM molecular dynamics simulations together with QM(DFT)/MM optimizations for key configurations have been performed to elucidate the enzymatic catalysis mechanism on the detoxification...Full Text Available

UK PubMed Central (United Kingdom)

A monensin-sensitive ruminal peptostreptococcus was able to grow rapidly (growth rate of 0.5/h) on an enzymatic hydrolysate of casein, but less than 23% of the amino acid nitrogen was ever utilized....Full Text Available

UK PubMed Central (United Kingdom)

Larvae of an anhydrobiotic insect, Polypedilum vanderplanki, accumulate very large amounts of trehalose as a compatible solute on desiccation, but the molecular mechanisms underlying...Full Text Available

UK PubMed Central (United Kingdom)

BackgroundLignocellulosic biomass such as wood is an attractive material for fuel ethanol production. Pretreatment technologies that increase the digestibility of cellulose and hemicellulose...Full Text Available

UK PubMed Central (United Kingdom)

Activatable or “smart” magnetic resonance contrast agents have relaxivities that depend on environmental factors such as pH or enzymatic activity, but the MR signal depends on...Full Text Available

UK PubMed Central (United Kingdom)

A rhabdomyosarcoma in a WAG/Rij rat with capacity for colony growth after tumour excision and enzymatic dissociation has been used to study response to high and low LET radiation. End points are tumor...Full Text Available

Science.gov (United States)

The goal of the present study was to describe the clinical, haematological and ultrasonographic findings and treatment of 17 cattle with pyelonephritis. Fifteen cattle had an abnormal general condition, which varied in severity; five animals had signs of colic. The urine was brownish-red in 11 animals and cloudy in 13. Clumps of purulent material were seen in the urine of nine animals and clots of blood in two. The specific gravity was lower than normal in 13 animals and ranged from 1.005 to 1.020. A urine test strip revealed protein in 16 animals, blood in 16 and leukocytes in 12. Bacteriological examination of urine yielded Corynebacterium renale in 11 animals, Arcanobacter pyogenes in two and Escherichia coli in one. Rectal examination revealed abnormalities of the urinary tract in 11 animals; there was dilatation of the left ureter and/or enlargement of the left kidney in eight cases, and dilatation of the right ureter and/or enlargement of the right kidney in ...

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The project was designed to solve the infra structural problem required for commercialization of food irradiation. In improvement of physical properties of corn starch, gamma irradiation was effective for increasing glucose productivity and for substituting traditional modified starches (acid modified starch, oxidized starch). In immobilization of microorganisms, the mass production method of natural red pigment was developed by using immobilized mold pellets. In Korean medicinal plants, 10 kGy gamma irradiation was effective for improving sanitary quality and increasing extraction yield. In evaluation of wholesomeness, gamma irradiated red ginseng could be safe on the genotoxic point of view. And also, six items of irradiated foods approved for human consumption from Korea ministry of health and welfare in May 19, 1995. 30 figs, 20 tabs, 54 refs. (Author).

British Library Electronic Table of Contents (United Kingdom)

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

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Efforts to assess and quantify deleterious effects from toxicants concentrate mainly on single agents whereas real world environmental and occupational exposures to natural and anthropogenic agents entail quite often the concomitant presence of several toxicants. For occupational important exposure situations, combined exposures to physical and chemical agents such as radon and smoking or asbestos and smoking, respectively, were shown to produce over-additive effects at exposure levels typical for earlier workplaces. Already the elucidation of possible health risks from a single agents, its dependence on exposure level, exposure rates, age at exposure and its expression in time is a complex endeavour. Therefore in the past and the present the main emphasis in radiation protection, toxicology, and public health is on the study and assessment of single toxicants. The existing data base on combined effect is rudimentary, mainly descriptive and rarely covers exposure ranges large enough to ...

British Library Electronic Table of Contents (United Kingdom)

Total synthesis of cis-solamin A was accomplished without using protecting groups starting from (-)-muricatacin in 11 steps with an overall yield of 4.5%. The backbone of cis-solamin A was constructed by olefin cross-metathesis between the tetrahydrofuran moiety and g-lactone moiety. An enzymatic kinetic transesterification procedure was successfully applied to the synthesis of an optically pure g-lactone moiety.

UK PubMed Central (United Kingdom)

This study evaluates the frequency and functions of immunocompetent T cells at the clonal level in solid human lung tissue versus peripheral blood. Enzymatic digestion of slices of histologically normal...Full Text Available

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This project has intended to develop alternative techniques to be used in food industry for food processing and utilization by safe irradiation methods. For improvement of rheology and processing in corn starch by irradiation, the production of modified starch with low viscosity as well as with excellent viscosity stability became feasible by the control of gamma irradiation dose levels and the amount of added inorganic peroxides to starch. Also, this project was developed the improvement methods of hygienic quality and long-term storage of dried red pepper by gamma irradiation. And, in Korean medicinal plants, 10 kGy gamma irradiation was effective for improving sanitary quality and increasing extraction yield of major components. For the sanitization of health and convenience foods, gamma irradiation was more effective than ozone treatment in decontamination of microorganisms, with minimal effect on the physicochemical properties analysed. In evaluation of wholesomeness, ...

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Managing sites contaminated with munitions constituents is an international challenge. Although the choice of approach and the use of Ecological Risk Assessment (ERA) tools may vary from country to country, the assurance of quality and the direction of ecotoxicological research are universally recognized as shared concerns. Drawing on a multidisciplinary team of contributors, 'Ecotoxicology of Explosives' provides comprehensive and critical reviews available to date on fate, transport, and effects of explosives. The book delineates the state of the science of the ecotoxicology of explosives, past, present, and recently developed. It reviews the accessible fate and ecotoxicological data for energetic materials (EMs) and the methods for their development. The chapters characterize the fate of explosives in the environment, then provide information on their ecological effects in key environmental media, including aquatic, sedimentary, and terrestrial habitats. The book ...

Science.gov (United States)

Progress is reported in several areas of research. The following cellulosic raw materials were selected for study: wheat, barley, and rice straws, rice hulls, sorghum, corn stover, cotton gin trash, newsprint, ground wood, and masonite steam-treated Douglas fir and redwood. Samples were collected, prepared, and analyzed for hexosans, pentosans, lignin, ash, and protein. Results of acid extraction and enzymatic hydrolysis are discussed. Yields of glucose, polyglucose, xylose, and arabinose are reported. Progress in process design and economic studies, as well as pilot plant process development and design studies, is summarized. (JGB)

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

Science.gov (United States)

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

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A sensitive radio-assay for ferritin was developed and used to examine serum ferritin levels in 105 patients with recurrent oral ulceration (ROU), 41 patients with Behcet's syndrome (BS), 42 with other ulcerative oral lesions, 35 patients with non-ulcerative oral lesions and in 78 controls. Ferritin levels increased with age and were significantly higher in males than females. The mean ferritin concentrations in male patients with ROU, BS or with other oral ulcers were significantly reduced in comparison with controls, and in female patients were significantly reduced in those with major aphthous ulcers. The prevalence of low serum ferritin levels was about 8% in patients with ROU, 15% in BS and 9.5% in patients with other ulcerative oral lesions, compared with less than 3% in patients with non-ulcerative oral disorders and in controls. Most of the iron-deficient patients were female. Serum ferritin levels did not directly correlate with serum iron levels and may be a more ...

Science.gov (United States)

The urine test strip is the most common test used to detect ketones in veterinary patients, but it can underestimate the degree of ketonuria and hence, ketonemia. Additionally, adequate urine samples for analysis may be difficult to obtain from dehydrated animals. The standard method used to detect and monitor ketonemia in human medicine is measurement of serum or whole blood beta-hydroxybutyrate (?HOB). A point-of-care (POC) analyzer has been validated for this purpose in humans. This study compared the accuracy of the POC device to an enzymatic reaction laboratory method for measurement of ?HOB in dogs. Although the POC sensor tended to overestimate ?HOB concentrations, there was good correlation (R(2) = 0.96) and good agreement between the 2 methods with a bias +/- precision of 0.0860 +/- 0.3410 mmol/L ?HOB. The POC ?HOB sensor can be useful for assessing ketonemia in dogs. PMID:21119867

British Library Electronic Table of Contents (United Kingdom)

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

British Library Electronic Table of Contents (United Kingdom)

SummaryBackground Intracellular transport via processive kinesin, dynein, and myosin molecular motors plays an important role in maintaining cell structure and function. In many cases, cargoes move distances longer than expected for single motors; there is significant evidence that this increased travel is in part due to multiple motors working together to move the cargoes. Although we understand single motors experimentally and theoretically, our understanding of multiple motors working together is less developed. Results We theoretically investigate how multiple kinesin motors function. Our model includes stochastic fluctuations of each motor as it proceeds through its enzymatic cycle. Motors dynamically influence each other and function in the presence of thermal noise and viscosity. We...

British Library Electronic Table of Contents (United Kingdom)

Recycling of carbon by the photorespiratory pathway involves enzymatic steps in the chloroplast, mitochondria, and peroxisomes. Most of these reactions are essential for plants growing under ambient CO2 concentrations. However, some disruptions of photorespiratory metabolism cause subtle phenotypes in plants grown in air. For example, Arabidopsis thaliana lacking both of the peroxisomal malate dehydrogenase genes (pmdh1pmdh2) or hydroxypyruvate reductase (hpr1) are viable in air and have rates of photosynthesis only slightly lower than wild-type plants. To investigate how disruption of the peroxisomal reduction of hydroxypyruvate to glycerate influences photorespiratory carbon metabolism we analyzed leaf gas exchange in A. thaliana plants lacking peroxisomal HPR1 expression. In addition, b...

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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.

British Library Electronic Table of Contents (United Kingdom)

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

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...

British Library Electronic Table of Contents (United Kingdom)

Teleost gut associated lymphoid tissue (GALT) consists of leucocyte populations located both intraepithelially and in the lamina propria with no structural organization. The present study aims to assess different protocols for the isolation of GALT cells from an important fish species in the Mediterranean aquaculture, the gilthead seabream. Mechanical, chemical and enzymatic treatments were assayed. Nylon wool columns and continuous density gradients were used for further separation of cell subpopulations. Light microscopy and flow cytometry showed that the highest density band (HD) consisted of a homogeneous lymphocytic population, whereas the intermediate density band (ID) corresponded to epithelial and secretory cells and some lymphocytes. Respiratory burst activity of total cell suspen...

British Library Electronic Table of Contents (United Kingdom)

Experiments based on a 23 central composite full factorial design were carried out in 200-ml stainless-steel containers to study the pretreatment, with dilute sulfuric acid, of a sugarcane bagasse sample obtained from a local sugar?alcohol mill. The independent variables selected for study were temperature, varied from 112.5?C to 157.5?C, residence time, varied from 5.0 to 35.0 min, and sulfuric acid concentration, varied from 0.0% to 3.0% (w/v). Bagasse loading of 15% (w/w) was used in all experiments. Statistical analysis of the experimental results showed that all three independent variables significantly influenced the response variables, namely the bagasse solubilization, efficiency of xylose recovery in the hemicellulosic hydrolysate, efficiency of cellulose enzymatic saccharificatio...

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Gas-insulated systems employing SF/sub 6/ are utilized in a variety of applications in power delivery systems. Because electric arcs, sparks or corona can decompose SF/sub 6/ into byproducts having chemical properties different from SF/sub 6/, environmental concerns arise regarding inadvertent human exposures to electrically-decomposed SF/sub 6/. Biological assays in our laboratory using mammalian cell culture systems have revealed that spark-decomposed SF/sub 6/ produces cell death, and the extent of cell killing increases with increasing spark energy. Chemical analysis of spark-decomposed SF/sub 6/ has identified the major decomposition pathways and byproducts, but biological testing of individual or byproduct mixtures has indicated that these major decomposition products do not contribute, in a significant fashion, to the cell lethality seen in our assays. Further experiments have suggested that S/sub 2/F/sub 10/ may be produced under our decomposition conditions and that this ...

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The eastern oyster (Crassostrea virginica) developed neoplastic disorders when experimentally exposed both in the laboratory and field to chemically contaminated sediment from Black Rock Harbor (BRH), Bridgeport, Connecticut. Neoplasia was observed in oysters after 30 or 60 days of continuous exposure in a laboratory flow-through system to a 20 mg/L suspension of BRH sediment plus postexposure periods of 3, 30, or 60 days. Composite tumor incidence was 13.6% for both exposures. Tumor occurrence was highest in the renal excretory epithelium, followed in order by gill, gonad, gastrointestinal, heart, and embryonic neural tissue. Regression of experimental neoplasia was not observed when the stimulus was discontinued. In field experiments, gill neoplasms developed in oysters, deployed in cages for 30 days at BRH and 36 days at a BRH dredge material disposal area in Central Long Island Sound, and kidney and gastrointestinal neoplasms developed in caged oysters deployed 40 days in Quincy ...

Energy Technology Data Exchange (ETDEWEB)

Research at the interface between biomolecules and inorganic nanocrystals has resulted in a great number of new discoveries. In part this arises from the synergistic duality of the system: biomolecules may act as self-assembly agents for organizing inorganic nanocrystals into functional materials; alternatively, nanocrystals may act as microscopic or spectroscopic labels for elucidating the behavior of complex biomolecular systems. However, success in either of these functions relies heavily uponthe ability to control the conjugation and assembly processes.In the work presented here, we first design a branched DNA scaffold which allows hybridization of DNA-nanocrystal monoconjugates to form discrete assemblies. Importantly, the asymmetry of the branched scaffold allows the formation of asymmetric2assemblies of nanocrystals. In the context of a self-assembled device, this can be considered a step toward the ability to engineer functionally distinct inputs and outputs.Next we develop an ...

Science.gov (United States)

An important advantage of carrying out enzymatic catalysis in organic media is the increased solubility of hydrophobic substrates. This study compares a model lipase catalyzed esterification of cholesterol using vinyl acetate (VA) in two such nontraditional media: high-pressure hexane and supercritical (SCF) ethane. The effect of using one of the reactants (VA) as a cosolvent to increase the solubility of the other reactant (cholesterol) in SCF ethane has been investigated. The thermodynamic activity of water (a(w)) in the reaction media was controlled by the direct addition of the salt hydrate pair Na(4)P(2)O(7)/Na(4)P(2)O(7).10H(2)O. The a(w) of the salt hydrate system is shown to be a function of pressure and its variation over the pressure range 104-173 bar has been estimated. The initial reaction rate in pressurized hexane was found to vary linearly with the cholesterol concentration. The reaction rate was also a function of pressure-the effect being more ...

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

Energy Technology Data Exchange (ETDEWEB)

The mechanisms involved in amino acid and sugar uptake by developing maize kernels were investigated. In the pedicel region of maize kernel, the site of nutrient unloading from phloem terminals, amino acids are accumulated in considerable amounts and undergo significant interconversion. A wide spectrum of enzymatic activities involved in the metabolism of amino acids is observed in these tissues. Subsequently, amino acids are taken up by the endosperm tissue in processes which require energy and the presence of carrier proteins. Conversely, no evidence was found that energy and carriers are involved in sugar uptake. This process of sugar uptake is not inhibited by metabolic inhibitors and shows nonsaturable kinetics, but the uptake is pH-dependent. L-glucose is taken up at a significantly reduced rate in comparison to D-glucose uptake. Based on analysis of radioactivity distribution among sugar fractions after incubations of kernels with radiolabeled D-glucose, it ...

Science.gov (United States)

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

Energy Technology Data Exchange (ETDEWEB)

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 enzymatic processes.

Science.gov (United States)

Symplastic transport occurs between neighbouring plant cells through functionally and structurally dynamic channels called plasmodesmata (PD). Relatively little is known about the composition of PD or the mechanisms that facilitate molecular transport into neighbouring cells. While transmission electron microscopy (TEM) provides 2-dimensional information about the structural components of PD, 3-dimensional information is difficult to extract from ultrathin sections. This study has exploited high-resolution scanning electron microscopy (HRSEM) to reveal the 3-dimensional morphology of PD in the cell walls of algae, ferns and higher plants. Varied patterns of PD were observed in the walls, ranging from uniformly distributed individual PD to discrete clusters. Occasionally the thick walls of the giant alga Chara were fractured, revealing the surface morphology of PD within. External structures such as spokes, spirals and mesh were observed surrounding the PD. ...

International Nuclear Information System (INIS)

Partially purified UDPgalactosyltransferase (EC 2.4.1.22) from bovine milk has been used to synthesize millimolar amounts of compounds such as Gal#beta#(1#->#4)Glc, Gal#beta#(1#->#4)GlcNAc-#beta#-hexanolamine, and Gal#beta#(1#->#4)-GlcNAc#beta#(1#->#4)GlcNAc. The same method has been used to prepare similar compounds containing "1"3C-enriched galactopyranosyl moieties. Gal#beta#(1#->#4)GlcNAc-#beta#-hexanolamine was also synthesized in a solid-phase system in which the GlcNAc-#beta#-hexanolamine glycoside was covalently linked to agarose beads. At pH 7.0 and at 1 to 5 mM Mn"2"+"+ the yields of the galactosyl saccharides are greater than 90% by using 10% excess of UDPGal donor. The use of a 90% enriched [1-"1"3C]galactosyl residue allowed the determination of the most abundant conformer about the galactopyranosyl-glycoside linkage by analysis of the carbon-carbon coupling constants from Cl to Gal to the C3', C4', and C5' of GlcNAc or Glc. 3 figures, 1 table.

Science.gov (United States)

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

DEFF Research Database (Denmark)

Citrate is an important naturally occurring inhibitor of calcium stone formation in urine. Urinary citrate excretion was examined in 43 consecutive patients with recurrent idiopathic calcium nephrolithiasis and in 50 normal controls by a specific enzymatic technique. Hypocitraturia (<1.6 mmol/24h) was found in 14 (33%) stone formers compared to 6 (12%) normal controls (p = 0.03). Citrate excretion expressed as citrate-creatinine ratio in 24 hour urine samples was significantly lower in the stone formers than in the healthy controls (p = 0.03), and significantly lower in healthy men, compared to healthy females (p = 0.006). There was a great variability in urinary citrate levels in both groups, and a considerable overlap in the urinary citrate excretion between normal subjects and stone formers. Factors other than urinary citrate excretion must therefore be of importance in the pathophysiology of calcium stone formation. Citrate-calcium ratio in urine proved to ...

Science.gov (United States)

We have constructed a 27-kDa hTERT C-terminal polypeptide (hTERTC27) devoid of domains required for telomerase activity and demonstrated that it is capable of nuclear translocation/telomere-end targeting. Here we showed that expression of a low level of hTERTC27 renders hTERT positive HeLa cells sensitive to H(2)O(2)-induced oxidative stress and subsequent cell senescence. The senescence-associated gene, the cyclin/cdk inhibitor p21(Waf1), was up-regulated. This occurs without changing the expression of endogenous hTERT, causing significant telomere shortening or inhibiting telomerase activity. Results from this study suggest for the first time that in addition to telomerase activity, the C-terminus of hTERT also plays a role in hTERT-mediated cellular resistance to oxidative stress. PMID:12565825

International Nuclear Information System (INIS)

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

International Nuclear Information System (INIS)

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

International Nuclear Information System (INIS)

Biodegradable and green plastics are the new tendency in the world. The effect of the electron beam irradiation in aromatic aliphatic copolyester and the blend with corn starch films (Ecoflex"R and Ecobras"R) were studied by tensile strength at break, elongation at break, Scanning Electronic Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FT-IR), Differential Scanning Calorimetry (DSC), crosslinking degree and biodegradability. The measurements in both, the machine direction and the transverse direction were made for mechanical tests. It was found that, the electron irradiation caused an increase in the strength at break of the blend with corn starch film, when doses of up to 10 kGy were applied. A significant decrease of the elongation at break of the blend with corn starch was observed at doses of 10 kGy and 40 kGy. It was not found important change in tensile properties for aromatic aliphatic copolyester. Structural changes of the samples (crosslinking or degradation) by ...

Science.gov (United States)

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

Science.gov (United States)

The natriuretic peptides are considered to be cardioprotective; however, their receptors have not been identified in human myocardium using radiolabeled analogs. Dendroaspis natriuretic peptide (DNP) has been recently identified as a new member of this peptide family and is thought to be less susceptible to enzymatic degradation. Therefore, we have developed the novel radiolabeled analog [125I]-DNP and used this to localize high-affinity (K(D)=0.2 nmol/L), saturable, specific binding sites in adult human heart (n=6) and coronary artery (n=8). In competition binding experiments, atrial natriuretic peptide and brain type natriuretic peptide had greater affinity for [125I]-DNP binding sites than C-type natriuretic peptide and the natriuretic peptide receptor (NPR)-C ligand, cANF. This rank order of potency suggested binding of [125I]-DNP was specific to NPR-A. Messenger RNA encoding NPR-A was identified in left ventricle and coronary artery smooth muscle, and ...

Energy Technology Data Exchange (ETDEWEB)

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

Energy Technology Data Exchange (ETDEWEB)

Previous studies have documented dissimilatory growth of bacteria on solid Mn{sup 4+} oxide, but Mn{sup 3+} oxides have not been previously studied; here the authors have demonstrated for the first time the bacterial reduction of manganite. Strain MR-4 of Shewanella putrefaciens was able to grow on and rapidly reduce insoluble needle-shaped crystals of synthetic manganite (MnOOH), converting them to soluble Mn{sup 2+} in the process. The rate of Mn{sup 3+} reduction was optimal at pH of 7.0 and 26 C consistent with an enzymatic reaction. In addition the rates of reduction were in proportion to the amount of manganite added, but nearly independent of the cell concentration present (e.g., cell number had only a small effect on the rate of Mn{sup 3+} reduction at early stages of growth) suggesting that surface properties were dictating the rates of metal reduction. This thesis was supported by major differences in reduction rates when Mn oxides of different surface ...

Energy Technology Data Exchange (ETDEWEB)

Phosphoenolpyruvate carboxykinase (GTP) (PEPCK) specifically utilizes a guanosine or inosine nucleotide as a substrate, yet it does not share extended sequence homology with other GTP-binding proteins, and the molecular basis for its nucleotide specificity is not understood. In an effort to locate the enzyme's nucleotide-binding site, the authors have studied the interaction of cytosolic PEPCK from rat liver with the photoprobe 8-azidoGTP, which fulfills the criteria of a specific photoaffinity label for PEPCK. The photoprobe binds reversibly to the enzyme prior to modification and at low concentrations causes greater than 60% inactivation-GTP provides nearly complete protection against inactivation by 8-azidoGTP, whereas phosphoenolpyruvate and metal ions provide partial protection. In addition, the photoprobe is a substrate for the enzyme and has a K{sub m} similar to that for GTP. However, the extent of covalent modification by ({sup 32}P)8-azido-GTP as measured by three ...

Energy Technology Data Exchange (ETDEWEB)

Partially purified UDPgalactosyltransferase (EC 2.4.1.22) from bovine milk has been used to synthesize millimolar amounts of compounds such as Gal..beta..(1..-->..4)Glc, Gal..beta..(1..-->..4)GlcNAc-..beta..-hexanolamine, and Gal..beta..(1..-->..4)-GlcNAc..beta..(1..-->..4)GlcNAc. The same method has been used to prepare similar compounds containing /sup 13/C-enriched galactopyranosyl moieties. Gal..beta..(1..-->..4)GlcNAc-..beta..-hexanolamine was also synthesized in a solid-phase system in which the GlcNAc-..beta..-hexanolamine glycoside was covalently linked to agarose beads. At pH 7.0 and at 1 to 5 mM Mn/sup 2 + +/ the yields of the galactosyl saccharides are greater than 90% by using 10% excess of UDPGal donor. The use of a 90% enriched (1-/sup 13/C)galactosyl residue allowed the determination of the most abundant conformer about the galactopyranosyl-glycoside linkage by analysis of the carbon-carbon coupling constants from Cl to Gal to the C3', ...

Energy Technology Data Exchange (ETDEWEB)

The processes of lung growth, injury, and repair are characterized by alterations in fibroblast synthesis and interstitial distribution of extracellular matrix components. Transforming growth factor beta (TGF-beta), which is postulated to play a role in modulating lung repair, alters the distribution of several matrix components such as collagen and fibronectin. We studied the effect of TGF-beta on the synthesis and distribution of the various glycosaminoglycans (GAGs) and whether these effects may explain its role in lung repair. Human diploid lung fibroblasts (IMR-90) were exposed to various concentrations of TGF-beta (0-5 nM) for variable periods of time (0-18 h). Newly synthesized GAGs were labeled with either (3H)glucosamine or (35S)sulfate. Individual GAGs were separated by size exclusion chromatography after serial enzymatic and chemical digestions and quantitated using scintillation counting. There was a dose-dependent increase in total GAG synthesis with ...

Energy Technology Data Exchange (ETDEWEB)

When a shortened enzymatic version of the Tetrahymena self-splicing intervening sequence (IVS) RNA is placed under catalytic conditions and irradiated at 254 nm, a covalent cross-link forms with high efficiency. The position of the cross-link was mapped by using three independent methods: RNase H digestion, primer extension with reverse transcriptase, and partial hydrolysis of end-labeled RNA. The cross-link is chemically unusual in that it joins two adenosines, A57 and A95. Formation of this cross-link depends upon the identity and concentration of divalent cations present and upon heat-cool renaturation of the IVS in a manner that parallels conditions required for optimal catalytic activity. Furthermore, cross-linking requires the presence of sequences within the core structure, which is conserved among group I intervening sequences and necessary for catalytic activity. Together these correlations suggest that a common folded structure permits cross-linking and ...

Energy Technology Data Exchange (ETDEWEB)

A robust fusion of the agricultural, industrial biotechnology, and energy industries can create a new strategic national capability for energy independence and climate protection. In his State of the Union Address (Bush 2006), President George W. Bush outlined the Advanced Energy Initiative, which seeks to reduce our national dependence on imported oil by accelerating the development of domestic, renewable alternatives to gasoline and diesel fuels. The president has set a national goal of developing cleaner, cheaper, and more reliable alternative energy sources to substantially replace oil imports in the coming years. Fuels derived from cellulosic biomass - the fibrous, woody, and generally inedible portions of plant matter - offer one such alternative to conventional energy sources that can dramatically impact national economic growth, national energy security, and environmental goals. Cellulosic biomass is an attractive energy feedstock because it is an abundant, domestic, renewable ...