salmonella microsomal assay: Topics by WorldWideScience.org

Determination of microsomal lauric acid hydroxylase activity by HPLC with flow-through radiochemical quantitation

Medicinal plants play a major role in the life of Turkish people and of late medicinal plant usage has increased in many countries. Green plants in general contain mutagenic and carcinogenic substances, but there is little information about the biological activities of herbal medicine. In the present study, therefore, various Turkish medicinal herbs were investigated for their genotoxic potential in the Salmonella typhimurium microsomal activation assay and the alkaline single cell gel electrophoresis (COMET) assay. Extracts from these medicinal herbs and some fractions of these extracts were examined. The species investigated were Arctium minus, Ecballium elatterium, Momordica charantia, Plantago major, Urtica dioica, Viscum album, Salvia triloba, Euphorbia rigida, Stachys lavandulifolia, Acteoside, Abies nordmannia. They are used for various immune disorders and are applied either topically or taken ...

1996-01-01

2

Past, present, and future of mutagens in cooked foods.

An assay for the microsomal hydroxylation of lauric acid (LA), based on HPLC with flow-through radiochemical detection, has been developed. Conditions were optimized for resolution and quantitation of three microsomal metabolites of "1"4C-LA, one of which has not been reported previously as a metabolite of LA in mammalian microsomal incubations. These products, 12-(omega)-hydroxy-LA, 11-(omega-1)-hydroxy-LA, and a novel metabolite, 10-(omega-2)-hydroxy-LA, were isolated by HPLC and identified by gas chromatography/mass spectrometry. In the presence of NADPH, the formation of all three metabolites was linear with time and microsomal protein concentration. Hydrogen peroxide also supported the microsomal metabolism of LA, although the ratio of metabolites was substantially different than that produced by NADPH-supported microsomes. Several ...

3

Direct detection of Salmonella without pre-enrichment in milk, ice-cream and fruit juice by PCR against hilA gene

Mutation assay with Salmonella typhimurium enabled us to detect various types of mutagens in cooked foods. A series of mutagenic heterocyclic amines has been isolated and identified in broiled fish...Full Text Available

1986-08-01

4

British Library Electronic Table of Contents (United Kingdom)

A novel PCR based assay was devised to specifically detect contamination of any Salmonella serovar in milk, fruit juice and ice-cream without pre-enrichment. This method utilizes primers against hilA gene which is conserved in all Salmonella serovars and absent from the close relatives of Salmonella. An optimized protocol, in terms time and money, is provided for the reduction of PCR contaminants from milk, ice-cream and juice through the use of routine laboratory chemicals. The simplicity, efficiency (time taken 3-4 h) and sensitivity (to about 5-10 CFU/ml) of this technique confers a unique advantage over other previously used time consuming detection techniques. This technique does not involve pre-enrichment of the samples or extensive sample processing, which was a pre-requisite in mos...

2012-01-01

5

Effect of Spices on hepatic microsomal enzyme function in mice

Induction of Hepatic Microsomal Cytochrome P450 by N,N-dimethylformamide in Sprague-Dawley Rats

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1984-06-01

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Increase in phorbol ester binding in liver microsomes after chronic administration of phenobarbital

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1999-03-01

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Negative Ames-test of cis-di(thiocyanato)-N,N'-bis(4,4'-dicarboxy-2,2'-bipyridine)Ru(II), the sensitizer dye of the nanocrystalline TiO{sub 2} solar cell

The effect of chronic administration of phenobarbital on the binding of phorbol-12,13-dibutyrate (({sup 3}H)PDBu), an activator of protein kinase C (PKC), was examined in rat liver microsomes. A significant increase in the number of binding sites was observed in microsomes of Fisher 344 rats. However, no change appeared in liver cytosol binding of PDBu. Consequently, a translocation process of PKC is unlikely. The increase in ({sup 3}H)PDBu binding in liver microsomes is significant 24 h. after one injection of phenobarbital and reaches its maximum in 2 days. In other strains of rats (ACI and lean Zucker), significant differences were found in the increase of ({sup 3}H)PDBu binding in microsomes. Fisher 344 were the most sensitive, lean Zucker rats, the least sensitive. Those results parallel the pentoxy-resorufin O demethylase activity in the microsomes of the same animals. EC{sub ...

1991-03-15

8

Sources of salmonellae in an uninfected commercially-processed broiler flock.

Dye-sensitized nanocrystalline TiO{sub 2} solar cells are currently under development. Since these cells contain an electrolyte solution we reviewed the health and safety aspects in view of indoor applications, where personal contact cannot be excluded. Only small amounts of chemicals are present in each cell and so there is no danger of acute toxicity. However, long-term effects often can be caused by incidental contact with minute amounts. For this reason we have tested cis-di(thiocyanato)-bis(4,4'-dicarboxy-2,2'-bipyridine)Ru(II), the sensitizer dye in the Ames test. The dye was not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.

2000-01-01

9

Flock infection and transport as sources of salmonellae in broiler chickens and carcasses.

Cultural monitoring was used to study the incidence and sources of salmonellae in a 4160 bird broiler flock during the growing period, transport and processing in a commercial plant. No salmonellae...Full Text Available

1980-07-01

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Farm-level risk factors for the presence of Salmonella in 89 Alberta swine-finishing barns

Cultural monitoring was used to determine the incidence and sources of salmonellae in a 4160-bird broiler flock raised on litter in 32 pens. Twenty-five of the pens remained apparently free of salmonellae...Full Text Available

1980-07-01

11

This study investigated potential risk factors for the presence of Salmonella on 89 Alberta swine-finishing farms with the use of a questionnaire. Salmonella status...Full Text Available

2007-10-01

12

CLINICOBACTERIOLOGICAL AND EPIDEMIOLOGICAL ...

In recent years, Salmonella enteritidis has become an increasingly important public health problem in Italy. In some parts of the country, the fraction of total human salmonella isolates...Full Text Available

1993-04-01

13

Transmission of Salmonella mbandaka to cattle from contaminated feed.

... Salmonella. Vdgella and Brucela also to the diagnosis of the septic- tylphoidal and enteric form of human peendotuber- auloals. ...

1963-11-01

14

The sensitivity of real-time PCR amplification targeting invasive Salmonella serovars in biological specimens

Salmonella mbandaka was isolated from cattle on three dairy farms. The duration of infection was less than four weeks and none of the animals became clinically ill. The animals had all consumed a diet...Full Text Available

1982-04-01

15

The relationships of salmonellae from infected broiler flocks, transport crates or processing plants to contamination of eviscerated carcases.

BackgroundPCR amplification for the detection of pathogens in biological material is generally considered a rapid and informative diagnostic technique. Invasive Salmonella...Full Text Available

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The associations between milk production, milk composition and Salmonella in the bulk milk supplies of dairy farms in Ontario.

Three flocks raised for broiler or roaster performance tests were studied to determine the incidence and sources of salmonellae during the growing period, transport and processing and to relate these...Full Text Available

1982-07-01

17

Spread of a Salmonella typhimurium Clone Resistant to Expanded-Spectrum Cephalosporins in Three European Countries

The purpose of this study was to assess changes in dairy herd milk production and milk composition associated with changes in Salmonella contamination of bulk milk on dairy farms in southwestern Ontario....Full Text Available

1989-04-01

18

Salmonellosis in Botswana

Twelve Salmonella typhimurium strains resistant to broad-spectrum cephalosporins were isolated from cases of gastroenteritis during 1996 to 1998 in Russia, Hungary, and Greece. Resistance...Full Text Available

1999-11-01

19

Salmonella enterica in Commercial Swine Feed and Subsequent Isolation of Phenotypically and Genotypically Related Strains from Fecal Samples?

Clinical salmonellosis in cattle is infrequently recorded in Botswana, the majority of salmonella isolations being made from apparently healthy animals. A salmonella carrier rate of 6·7% was...Full Text Available

1971-09-01

20

Roles of curli, cellulose and BapA in Salmonella biofilm morphology studied by atomic force microscopy

The purpose of this study was to determine the occurrence and genotypic relatedness of Salmonella enterica isolates recovered from feed and fecal samples in commercial swine production...Full Text Available

2010-11-01

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Resuscitation of Salmonella enterica Serovar Typhimurium and Enterohemorrhagic Escherichia coli from the Viable but Nonculturable State by Heat-Stable Enterobacterial Autoinducer

BackgroundCurli, cellulose and the cell surface protein BapA are matrix components in Salmonella biofilms. In this study we have investigated the roles of these...Full Text Available

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Phosphorylated PmrA Interacts with the Promoter Region of ugd in Salmonella enterica Serovar Typhimurium

Salmonella enterica serovar Typhimurium and enterohemorrhagic Escherichia coli were stressed by prolonged incubation in water microcosms until it was no longer possible...Full Text Available

2002-10-01

23

Molecular Analyses of Salmonellaenterica Isolates from Fish Feed Factories and Fish Feed Ingredients

The Salmonella PmrA-PmrB system controls the expression of genes necessary for polymyxin B resistance. Four loci were previously identified as part of the regulon, and interaction of...Full Text Available

2000-07-01

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High Prevalence of Extended-Spectrum Beta Lactamases among Salmonella enterica Typhimurium Isolates from Pediatric Patients with Diarrhea in China

Isolates of the most commonly observed salmonella serovars in Norwegian fish feed factories from 1998 to 2000 (Salmonella enterica serovar Agona, S. enterica...Full Text Available

2003-02-01

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Compounds Which Serve as the Sole Source of Carbon or Nitrogen for Salmonella typhimurium LT-2

We investigated the extended-spectrum beta lactamases among 62 Salmonella enterica Typhimurium isolates recovered from children with diarrhea in a Chinese pediatric hospital. A large...Full Text Available

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Comparison of Genotypes of Salmonella enterica Serovar Enteritidis Phage Type 30 and 9c Strains Isolated during Three Outbreaks Associated with Raw Almonds?

About 600 compounds were screened as possible carbon or nitrogen sources for Salmonella typhimurium LT-2. About 100 utilizable compounds were found.

1969-10-01

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Attenuated Salmonella typhimurium reduces ovalbumin-induced airway inflammation and T-helper type 2 responses in mice

In 2000 to 2001, 2003 to 2004, and 2005 to 2006, three outbreaks of Salmonella enterica serovar Enteritidis were linked with the consumption of raw almonds. The S....Full Text Available

2010-06-01

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A Murine Model to Study the Antibacterial Effect of Copper on Infectivity of Salmonella Enterica Serovar Typhimurium

Cytokines produced by Th2 cells are responsible for the pathogenesis of asthma. Th1-biased immune responses caused by attenuated salmonella have the potential to relieve asthmatic symptoms. We evaluated...Full Text Available

2006-07-01

29

Invasiveness and persistence of Salmonella enteritidis, Salmonella typhimurium, and a genetically defined S. enteritidis aroA strain in young chickens.

This study investigated the effect of copper as an antibacterial agent on the infectivity of Salmonella enterica serovar Typhimurium. Mice were infected orally with a standardized dose...Full Text Available

2011-01-01

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Effect of paraquat on microsomal lipid peroxidation in vitro and in vivo. [Rats, rabbits, man, mice

Newly hatched chicks were dosed orally with a Salmonella typhimurium wild-type strain, an S. enteritidis wild-type strain, and a genetically defined S. enteritidis aroA vaccine candidate, strain CVL30....Full Text Available

1994-11-01

31

Safety evaluation of Propionibacterium freudenreichii ET-3 culture.

Rat lung and liver microsomes did not undergo lipid peroxidation in the absence of iron when incubated with NADPH and concentrations of paraquat ranging from 10/sup -7/ to 10/sup -2/ M. Paraquat also did not stimulate rat liver and lung microsomal peroxidation induced by added iron and NADPH, and was inhibitory at concentrations above 10 ..mu..M. Similarly, no stimulation of peroxidation was produced by paraquat in rabbit or human lung microsomes; however, under similar conditions, paraquat enhanced NADPH/iron-dependent peroxidation in mouse lung and liver microsomes obtained from rats sacrificed at 12, 18, and 24 hr following a lethal dose of paraquat (50 mg/kg, ip), there was no loss of vitamin E or increase in susceptibility to in vitro peroxidation which would be expected if lipid peroxidation had occurred in vivo although extensive lung damage developed during this time period. These results ...

1980-01-01

32

Presence of Arsenic Resistance in Salmonella enterica Serovar Kentucky and Other Serovars Isolated from Poultry

Propionibacterium freudenreichii ET-3 culture, a cell-free product of whey fermentation using P. freudenreichii ET-3 (7025), has been shown to promote the growth of Bifidobacteria through the action of 1,4-dihydroxy-2-naphthoic acid (DHNA), and therefore, has potential use in the food and supplement industries. Although currently used as a food ingredient in Japan, the safety of this novel ingredient has not been previously evaluated through traditional toxicity testing. Therefore, here we report the results of standard toxicological testing performed on P. freudenreichii ET-3 culture. In a 4-week oral toxicity study, administration of 6000mg/kg body weight/day P. freudenreichii ET-3 culture was without compound-related adverse effects on clinical signs, body weights, food consumption, ophthalmology, hematology, clinical chemistry, urinalysis, organ weights, and gross and microscopic findings in male and female Sprague-Dawley rats. Furthermore, in vitro mutagenicity testing ...

2011-03-22

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A Response Surface Model Based on Absorbance Data for the Growth Rates of Salmonella enterica Serovar Typhimurium as a Function of Temperature, NaCl, and pH

... B. P. Rosen. An arsenic metallochaperone for an arsenic detoxification pump. Proc. Natl. Acad. Sci. U. S. A 103( ... ...

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

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2007-04-01

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Abnormalities in the microsomal oxidases of the WHO standard reference strain of Musca domestica*

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

36

Transfer of Plasmid-Mediated CTX-M-9 from Salmonella enterica Serotype Virchow to Enterobacteriaceae in Human Flora-Associated Rats Treated with Cefixime?

Observations made during biochemical and toxicological studies of the housefly, in which the WHO standard reference (SR) strain was used as a standard, indicated that this strain differs from other...Full Text Available

1975-01-01

37

Chromosomal Integration of the Extended-Spectrum ?-Lactamase Gene bla_CTX-M-15 in Salmonella enterica Serotype Concord Isolates from Internationally Adopted Children?

Food animals are a potential source of CTX-M resistance genes for humans. We evaluated the transfer of the bla_CTX-M-9 gene from an animal strain of Salmonella enterica...Full Text Available

2010-01-01

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Association between Handling of Pet Treats and Infection with Salmonella enterica Serotype Newport Expressing the AmpC ?-Lactamase, CMY-2

We report the emergence of Salmonella enterica isolates of serotype Concord (and its monophasic variant 6,7:l,v:-) producing the extended-spectrum β-lactamases (ESBLs) SHV-12...Full Text Available

2009-05-01

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Antimicrobial resistance, virulence-associated genes, and pulsed-field gel electrophoresis profiles of Salmonella enterica subsp. enterica serovar Typhimurium isolated from piglets with diarrhea in Korea

Resistance to the extended-spectrum cephalosporins can occur in Salmonella species via the production of extended-spectrum and AmpC β-lactamases. We describe human infections...Full Text Available

2003-10-01

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METHOD FOR ASSAYING PROTEASE ACTIVITY USING POLYMER MEMBRANE

Salmonella enterica subsp. enterica serovar Typhimurium was isolated from diarrheic piglets in 2 periods, 2000–2001 (n = 25) and 2005–2006...Full Text Available

2011-01-01

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J-STORE (Japan)

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2007-09-14

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Assays for measuring arachidonic acid release from phospholipids

Altered lauric acid metabolism in renal microsomes from spontaneously hypertensive rats (SHR)

No abstract available.

1991-01-01

43

Radiation inactivation target size of rat adipocyte glucose transporters in the plasma membrane and intracellular pools

Studies investigated whether changes in omega- and (omega-1)-hydroxylation (OH) of lauric acid (LA) occurred in renal microsomes prepared from SHR compared to Wistar-Kyoto (WK) control rats. Systolic blood pressure in age-matched adult SHR and WKR were 189 +/- 3 and 123 +/- 4 mm Hg(anti X +/- SE) respectively (p < 0.001). No significant differences between SHR and WKR were seen in body weight, kidney weight or renal microsomal protein content. Renal microsomes, prepared from whole kidneys, were incubated with 10 mM NADPH and ["1"4C]LA at concentrations between 5-50 #mu#M. The 11- and 12-OH metabolites of LA were separated by HPLC using a reverse phase column with a methanol:water:acetic acid (62:37.8:0.2) mobile phase. Apparent (app) V/sub max/ values for 12-OH in WKR and SHR were 0.87 +/- 0.19 vs 1.48 +/- .11 nmoles/mg protein/min (p < 0.05), respectively, while values for 11-OH were 0.51 +/- 0.12 vs 0.60 +/- .07, ...

1986-04-13

44

Lipid composition of liver microsomes and mitochondria after acute and chronic {gamma}-irradiation of rats

The in situ assembly states of the glucose transport carrier protein in the plasma membrane and in the intracellular (microsomal) storage pool of rat adipocytes were assessed by studying radiation-induced inactivation of the D-glucose-sensitive cytochalasin B binding activities. High energy radiation inactivated the glucose-sensitive cytochalasin B binding of each of these membrane preparations by reducing the total number of the binding sites without affecting the dissociation constant. The reduction in total number of binding sites was analyzed as a function of radiation dose based on target theory, from which a radiation-sensitive mass (target size) was calculated. When the plasma membranes of insulin-treated adipocytes were used, a target size of approximately 58,000 daltons was obtained. For adipocyte microsomal membranes, we obtained target sizes of approximately 112,000 and 109,000 daltons prior to and after insulin treatment, ...

1987-06-15

45

Mutagenic and cytotoxic activity of doxorubicin and daunorubicin derivatives on prokaryotic and eukaryotic cells.

Acute {gamma}-irradiation of rats at doses of 100 and 270 Gy stimulates lipid synthesis and changes the lipid composition of liver cell organelles. The content of cholesterol and cholesterol esters in microsomes increased at 100 Gy and decreased at 270 Gy, with total phospholipid content remaining unchanged. The lipid content in mitochondria decreased considerably 1 h after irradiation at 270 Gy. This change was significantly less pronounced 47 h later. Under chronic {gamma}-irradiation (0.129 Gy/day), cholesterol and cardiolipin in mitochondria increased. The changes in lipid content caused by acute irradiation are presumed to be related to activated synthesis of lipids in the liver. The modification of the lipid content of mitochondria observed in chronically irradiated rats may indicate that energy-metabolizing liver cell systems are involved in the adaptation to irradiation.

1994-07-01

46

Evaluation of the Widal tube agglutination test for the diagnosis of typhoid fever among children admitted to a rural hdospital in Tanzania and a comparison with previous studies

The mutagenic and cytotoxic activity of two newly synthesized doxorubicin derivatives and of one daunorubicin derivative were studied in V79 Chinese hamster cells and bacteria (Salmonella typhimurium...Full Text Available

1984-07-01

47

Changes in Tetracycline Susceptibility of Enteric Bacteria following Switching to Nonmedicated Milk Replacer for Dairy Calves?

BackgroundThe diagnosis of typhoid fever is confirmed by culture of Salmonella enterica serotype Typhi (S. typhi). However, a more rapid, simpler,...Full Text Available

48

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

A randomized intervention study was conducted to determine if discontinuing use of calf milk replacer medicated with oxytetracycline results in increased tetracycline susceptibility in Salmonella...Full Text Available

2008-06-01

49

Adrenaline modulates the global transcriptional profile of Salmonella revealing a role in the antimicrobial peptide and oxidative stress resistance responses

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

1985-05-01

50

beta-Lactamases among extended-spectrum beta-lactamase (ESBL)-resistant Salmonella from poultry, poultry products and human patients in The Netherlands

BackgroundThe successful interaction of bacterial pathogens with host tissues requires the sensing of specific chemical and physical cues. The human gut contains a huge number of...Full Text Available

51

DEFF Research Database (Denmark)

Objectives: The purpose of this work was to study the genetic determinants responsible for extended-spectrum beta-lactamase (ESBL) resistance of Salmonella isolated from Dutch poultry, poultry meat and hospitalized humans. Methods: Thirty-four ESBL-resistant Salmonella isolates from The Netherlands were tested towards 21 antimicrobial agents. PCR and sequencing were used to determine the underlying genetic determinants responsible for the ESBL phenotypes. The transferability of the ESBL phenotypes was tested by conjugation to a susceptible Salmonella enterica serovar Dublin and plasmid purification, restriction fragment length polymorphism (RFLP) and pulsed-field gel electrophoresis (PFGE) were employed to further characterize a subset of the isolates. Results: A great genetic diversity was seen among the isolates. The bla(TEM-52) gene was most predominant and was found among Salmonella enterica ...

2005-01-01

52

Microsomal Electron Transfer in Higher Plants: Cloning and Heterologous Expression of NADH-Cytochrome b₅ Reductase from Arabidopsis

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

AtCBR, a cDNA encoding NADH-cytochrome (Cyt) b₅ reductase, and AtB5-A and AtB5-B, two cDNAs encoding Cyt b₅, were isolated from Arabidopsis....Full Text Available

1999-01-01

53

In vitro assay for curative activity in blood of mice infected with Trypanosoma musculi.

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 than succinate or ATP-driven Ca/sup 2 +/ ...

1984-04-01

54

Detection and enumeration of toxin-producing Pasteurella multocida with a colony-blot assay.

An in vitro assay for curative antibody present in plasma of mice cured of Trypanosoma musculi is described. The assay involves the addition of plasma to a sample of infected blood, followed by hourly...Full Text Available

1986-08-01

55

Comparison of a Commercial Multiplex Real-Time PCR to the Cell Cytotoxicity Neutralization Assay for Diagnosis of Clostridium difficile Infections?

Colonies of toxin-producing Pasteurella multocida were detected with peroxidase-labeled monoclonal antibodies by a membrane assay. Examination of the specificity of the assay with 29 P. multocida cultures...Full Text Available

1991-07-01

56

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

A commercial multiplex real-time PCR assay (Cepheid Xpert C. difficile assay) for the diagnosis of Clostridium difficile infection was evaluated. The sensitivity and...Full Text Available

2009-11-01

57

Prostaglandin synthesis by chicken and rat lung microsomes

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

58

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

A comparison between chicken and rat lung was made for microsomal prostaglandin (PG) synthesis from 1-/sup 14/C-arachidonic acid. Microsomal protein (2.0 mg) from chicken or rat lung was incubated in the presence of 20 ..mu..g of 1-/sup 14/C-arachidonic acid (specific activity = 3 x 10/sup 6/ dpm/..mu..mol for chicken; 6 x 10/sup 6/ dpm/..mu..mol for rat), 0.05 M Tris-HCl buffer (pH = 8.0), 0.5 mM epinephrine, and 1 mM reduced glutathione in a total volume of 0.5 ml in a 37/sup 0/C water bath with shaking for 15 min. After acidification with 1 M HCl to pH 3, prostaglandins were extracted with ethyl acetate. The products of the reactions were separated by reversed phase chromatography, and the radioactivity of each prostanoid fraction was determined. The predominant prostanoid synthesized by chicken lung microsomes was PGE/sub 2/, followed by much lower amounts of thromboxane B/sub 2/ (TXB/sub 2/), PGF/sub 2//sub ..cap ...

1986-03-01

59

Comparison of Conditions for Cell Death-Inducing Agents Using a High Throughput-Compatible Nuclear Staining Assay

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

1990-01-01

60

Comparison between DNA damage investigated by the ''Comet'' assay and chromosomal damage induced in human lymphocytes by radiation and chemicals

Full Text Available

2008-09-01

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61

Avian LH and FSH: comparison of several radioimmunoassays

Short communication.

1995-08-20

62

Salmonella radicidation of poultry carcasses

Comparisons were made between various LH and FSH radioimmunoassays currently being used to measure avian hormones. The two LH assays were the homologous chicken system of Follett et al. (1972) and the turkey assay of Burke et al. (1979). These assays were also used in heterologous arrangement by interchanging the iodinated LH fractions and antisera. Five FSH assays were analyzed: two homologous chicken systems (Scanes et al., 1977; Sakai and Ishii, 1980) an assay based on mammalian materials (rat FSH and anti-ovine FSH antiserum, and one using labelled turkey FSH (Burke et al., 1979) with an anti-chicken FSH antiserum. The potencies of purified chicken and turkey gonadotrophin preparations and of a range of plasma samples from Japanese quail were measured in each assay. The two LH systems showed some degree of species specificity, such that chicken LH was more ...

1983-04-01

63

Activity of Citrus Essential Oils against-Escherichia coli-O157:H7 and-Salmonella-spp. and Effects on Beef Subprimal Cuts under Refrigeration

This thesis reports investigations using gamma-radiation to decontaminate poultry carcasses. The application to foods of doses of ionizing radiation sufficient to reduce the number of viable specific non-sporeforming pathogenic microorganisms so that none is detectable in the treated food by any standard method is termed radicidation. The doses used in this study were at such a level that no undesirable or unfavourable side-effects occurred. The effects of these doses were studied on salmonellae and other microorganisms present in, or associated with poultry carcasses and in liquid and on solid culture media as well. Decimal reduction (D_1_0) values were estimated. These represent the dose (kGy) required to achieve a reduction in initial colony count from N_0 to 0.1 N_0. Together with the estimation of the numbers of Salmonella present per carcass the data were used to predict the effect of an ionizing radiation treatment of poultry. Data on ...

2005-09-22

64

British Library Electronic Table of Contents (United Kingdom)

Abstract:- Escherichia coli-O157:H7 and-Salmonella-spp. are bacterial pathogens often associated with beef, and cause many cases of foodborne illness each year in the United States. During beef slaughter and processing, these bacteria may spread from the hide or intestines to the carcass. The objective of this research was to investigate the use of naturally occurring compounds citrus essential oils (CEOs) extracted from orange peel to reduce or eliminate these pathogens at the chilling stage of processing, or during fabrication. Brisket flats (used to simulate beef subprimals) were spot inoculated with approximately 6 log of surrogate generic-E. coli-cocktail (previously shown to be identical in growth and survival parameters to-E. coli-O157:H7 and-Salmonella-spp.). Following drying, CEOs...

2011-01-01

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High hydrostatic pressure processing reduces Salmonella enterica serovars in diced and whole tomatoes

British Library Electronic Table of Contents (United Kingdom)

Fresh and fresh-cut tomatoes have been associated with numerous outbreaks of salmonellosis in recent years. One effective post harvest treatment to reduce Salmonella enterica in tomatoes may be high pressure processing (HPP). The objectives of the study were to determine the potential for HPP to reduce S. enterica serovars Newport, Javiana, Braenderup and Anatum in tryptic soy broth (TSB) and to determine the effect of HPP to reduce the most pressure resistant of the four serovars from fresh diced and whole tomatoes. To evaluate pressure resistance, TSB containing 8 log CFU/ml of one of the four serovars was packaged in sterile stomacher bags and subjected to one of three different pressures (350, 450 or 550MPa) for 120s. The most pressure resistant S. enterica serovar evaluated was Braend...

2011-01-01

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Biomagnetic separation of Salmonella Typhimurium with high affine and specific ligand peptides isolated by phage display technique

Suppression of receptors for prolactin and estrogen in rat liver due to treatment with the growth hormone analogue produced by the tapeworm Spirometra mansonoides.

Analyses of food-borne pathogens are of great importance in order to minimize the health risk for customers. Thus, very sensitive and rapid detection methods are required. Current conventional culture techniques are very time consuming. Modern immunoassays and biochemical analysis also require pre-enrichment steps resulting in a turnaround time of at least 24 h. Biomagnetic separation (BMS) is a promising more rapid method. In this study we describe the isolation of high affine and specific peptides from a phage-peptide library, which combined with BMS allows the detection of Salmonella spp. with a similar sensitivity as that of immunomagnetic separation using antibodies.

2007-04-15

67

Inactivation of aflatoxin B1 by using the synergistic effect of hydrogen peroxide and gamma radiation

Somatogenic hormones play an important role in regulation of receptors for prolactin (PRL) and estrogen. Plerocercoids of the tapeworm, S. mansonoides produce a factor which mimics some, but not all of the actions reported for GH. Intact female rats were subjected to a constant infusion of plerocercoid growth factor (PGF) via a subcutaneous infection for two weeks to determine if PGF influences receptors for PRL, GH or estradiol. The rate of weight gain in the PGF-treated rats was accelerated in spite of a marked reduction in serum GH. Partially-purified PGF specifically displaced [125I]hGH from rat liver receptors but microsomes prepared from rats treated with PGF specifically bound significantly less [125I]hGH than microsomes from control rats. The reduction in [125I]hGH binding was not due to occupancy or to a change in affinity but to a suppression in receptor concentration. Scatchard analysis of [3H]estradiol binding in rat liver cytosols ...

1986-01-01

68

Carbon tetrachloride-induced release of calcium from isolated hepatocytes

Inactivation of aflatoxin B1 was studied by using gamma radiation and hydrogen peroxide. A 100-krad dose of gamma radiation was sufficient to inactivate 50 micrograms of aflatoxin B1 in the presence of 5% hydrogen peroxide, and 400 krad was required for total degradation of 100 micrograms of aflatoxin in the same system. Degradation of aflatoxin B1 was confirmed by high-pressure liquid chromatographic and thin-layer chromatographic analysis. Ames microsomal mutagenicity test showed loss of aflatoxin activity. This method of detoxification also reduces the toxin levels effectively in artificially contaminated groundnuts.

1989-02-01

69

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

Studies have shown that CCl_4 administration to rats inhibits endoplasmic reticulum calcium pump activity and reduces the amount of calcium associated with subsequently isolated microsomal subcellular fractions. This report confirms that exposure of isolated hepatocytes to CCl_4 rapidly produces these effects in isolated parenchymal cells and demonstrates that when isolated hepacytes are exposed to CCl_4 calcium is rapidly released from cells. This release can be detected with a calcium ion-selective electrode when cells are incubated in a medium with low extracellular calcium. Calcium released from an intracellular pool(s) may initiate hepatotoxic changes in liver. (author).

70

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

Three cDNAs, designated IIA3, IIA3v, and IIA4, coding for P450s in the CYP2A gene subfamily were isolated from a {lambda}gt11 library prepared from human hepatic mRNA. Only three nucleotide differences and a single amino acid difference, Leu{sup 160}{yields}His, were found between IIA3 and IIA3v, indicating that they are probably allelic variants. IIA4 displayed 94% amino acid similarity with IIA3 and IIA3v. The three cDNAs were inserted into vaccinia virus, and recombinant viruses were used to infect human hepatoma Hep G2 cells. Only IIA3 was able to produce an enzyme that had a reduced CO-bound spectrum with a {lambda}{sub max} at 450 nm. This expressed enzyme was able to carry out coumarin 7-hydroxylation and ethoxycoumarin O-deethylation. cDNA-expressed IIA3v and IIA4 failed to incorporate heme and were enzymatically inactive. Analysis of IIA proteins in human liver microsomes, using antibody against rat IIA2, revealed two proteins of 49 and 50 kDa, the former ...

1990-02-06

71

Assay of the uranium and plutonium content in process residues and wastes using the correction for sample self attenuation in segmented gamma scanning system

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 ...

1990-02-01

72

Tests for mutagencity of free radicals formed in irradiated sugars and amino acids

A method of the correction for sample self attenuation used in segmented gamma scanner (SGS) is described. The method of calibration for assaying the uranium and plutonium contents of each category of heterogeneous process residues and wastes in nuclear facilities is studied. The effect of variable measurement parameter on measurement results is also studied. The measurement results of SGS assay is compared with that of the destructive assay (DA), which is aimed at evaluating SGS method for assaying uranium and plutonium content in process residues and wastes. The deviation of two assay results is 3.6%. The SGS assay results and DA assay results both are coincided well in error limits. Four category of heterogeneous process residues and wastes in nuclear facilities have assayed successfully in physical inventory. The ...

1998-09-01

73

Salmonella enterica Serovar Typhimurium Expressing a Chromosomally Integrated Copy of the Bacillus anthracis Protective Antigen Gene Protects Mice against an Anthrax Spore Challenge

Radicals formed in gamma-irradiated crystals of galactose and glycine were found, upon dissolution, to cause mutagenesis of Salmonella typhimurium strains TA-98 and TA-100. Although the reproducibility of the results has not been adequately determined, they suggest the possibility of developing a test to measure the mutagenic-carcinogenic potential of radiation-induced free radicals with a microbial system.

1977-05-01

74

Platelet-activating factor mediates hemodynamic changes and lung injury in endotoxin-treated rats.

Protective immunity against infection with Bacillus anthracis is almost entirely based on a response to the protective antigen (PA), the binding moiety for the two other toxin components....Full Text Available

2003-07-01

75

Destruction of Salmonellae on poultry meat with lysozyme, EDTA, x ray, microwave, and chlorine

Within 20 min after intraperitoneal injection of Salmonella enteritidis endotoxin in rats, blood platelet-activating factor (PAF) increased from 4.3 +/- 1.3 to 13.7 +/- 2.0 ng/ml (P less than 0.01)...Full Text Available

1987-05-01

76

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

Lysozyme, ethylenediaminetetracetic acid, chlorine, x-irradiation and microwaves were used in experimental attempts to eliminate Salmonella senftenberg 775W or Salmonella typhimurium from turkey drumsticks and whole carcasses. Turkey drumsticks or whole carcasses were artificially contaminated with S. senfenberg 775W or S. typhimurium in concentrations ranging between 5 x 10"5 to 8 x 10"5 viable cells per ml. of contaminating fluid. After each treatment, samples were cultured, plated, and tested according to standard methods to determine the susceptibility of Salmonella organisms to the particular treatment. A 0.1 percent solution of lysozyme eliminated the S. senftenberg 775W at 22"0C within three hours. A 0.5 percent solution of ethylenediaminetetracetic acid failed to destroy the test organism under the same conditions. Eighty thousand rads of X-ray eliminated the test organism on turkey drumsticks but failed to remove ...

77

Blockade of catecholamine-induced growth by adrenergic and dopaminergic receptor antagonists in Escherichia coli O157:H7, Salmonella enterica and Yersinia enterocolitica

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

2009-08-01

78

A rapid and simple DNA extraction procedure to detect Salmonella spp. and Listeria monocytogenes from fresh produce using real-time PCR

BackgroundThe ability of catecholamines to stimulate bacterial growth was first demonstrated just over a decade ago. Little is still known however, concerning the nature of the putative...Full Text Available

79

Some New Aspects of the in Vivo Assay for Nitrate Reductase in Wheat (Triticum aestivum L.) Leaves

DNA isolation procedures significantly influence the outcome of PCR-based detection of human pathogens. Unlike clinical samples, DNA isolation from food samples such as fresh and fresh-cut produce has remained a formidable task and has hampered the sensitivity and accuracy of molecular methods. We...

80

Processing and microfiltration of mosquitoes for malaria antigen detection in a rapid dot immunobinding assay.

Experiments were carried out to clarify problems encountered in measuring metabolic and storage pool sizes of nitrate in wheat leaf sections with an in vivo nitrate reductase assay....Full Text Available

1980-01-01

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81

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

Data on a technique for the detection of antigen from arthropod vectors in a dot immunobinding assay are presented. In this system, antigen present in the vector was first solubilized in sodium dodecyl...Full Text Available

1990-08-01

82

Nitric Oxide Emissions from Soybean Leaves during in Vivo Nitrate Reductase Assays ¹

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

1994-11-01

83

Los Alamos second-generation system for passive and active neutron assays of drum-size containers

Recent work identified acetaldehyde oxime as the predominant product purged by inert gases from anaerobic in vivo nitrate reductase (NR) assays of soybean (Glycine max...Full Text Available

1987-09-01

84

Improved Susceptibility Disk Assay Method Employing an Agar Overlay Technique

We describe in a comprehensive fashion the Los Alamos second-generation system for passive and active neutron assays of drum-size containers. The developmental history of this 7-year project is presented with emphasis on the pulsed active neutron technique (differential dieaway), which has achieved milligram levels of assay sensitivity for both plutonium and uranium wastes. We describe in detail the matrix effects for both passive and active neutron assays. We present in a thorough fashion our novel approach to achieving comprehensive corrections for these matrix effects using measurements made during the assays. We develop a matrix correction formalism based on separate neutron absorption and moderator indices determined from these measurements. These are presented as a series of analytic functions fitted to the data. Absolute calibrations and calibration standards are discussed, as is a practical ...

1972-09-17

85

Immunoradiometric measurement of the factor VIII procoagulant antigen.

Susceptibility disks impregnated with 35 different antibiotics were assayed by the current Code of Federal Regulations method and by a new method employing an agar overlay. The new...Full Text Available

1978-11-01

86

Experimental Differential Light-Scattering Correction to the Circular Dichroism of Bacteriophage T2

A fluid-phase immunoradiometric assay has been developed which identifies an antigen on the Factor VIII (antihemophilic factor) procoagulant protein. This sensitive and quantitative assay is not influenced...Full Text Available

1978-11-01

87

Evaluation of Direct Detection of Mycobacterium tuberculosis Rifampin Resistance by a Nitrate Reductase Assay Applied to Sputum Samples in Cotonou, Benin?

Experimental techniques are presented that can be used to assay and correct for differential light scattering effects in circular dichroism spectra of biological macrostructures. The assay is based...Full Text Available

1973-01-01

88

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

The aim of this study was to evaluate a nitrate reductase assay (NRA) performed on smear-positive sputa for the direct detection of rifampin resistance in Mycobacterium tuberculosis....Full Text Available

2007-07-01

89

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

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

2001-11-01

90

Consistency in potency assay of tetanus toxoid in mice

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

1988-10-01

91

Assay for the enzymatic conversion of indoleacetic acid to 3-methylindole in a ruminal Lactobacillus species.

The use of mice for the assay of tetanus toxoids would offer considerable advantages over the use of guinea-pigs, but mice cannot readily be immunized with the fluid tetanus toxoid at present designated...Full Text Available

1959-01-01

92

Acetaldehyde Oxime, A Product Formed during the In Vivo Nitrate Reductase Assay of Soybean Leaves 1

An assay to measure the rate of enzymatic formation of 3-methylindole (3MI) from indoleacetic acid (IAA) in Lactobacillus sp. strain 11201 was developed. The reaction mixture contained 50 micrograms...Full Text Available

1990-03-01

93

A duplex real-time RT-PCR assay for the detection of California serogroup and Cache Valley viruses

Evolution of nitrogen oxides (NO_(x), primarily as nitric oxide) from soybean (Glycine max [L.] Merr.) leaves during purged in vivo nitrate reductase assays...Full Text Available

1984-09-01

94

Diagnosing Pulmonary Aspergillosis in Patients With Hematological Malignancies: A Multicentre Prospective Evaluation of an Aspergillus PCR Assay and a Galactomannan ELISA in Bronchoalveolar Lavage Samples

A duplex TaqMan real-time RT-PCR assay was developed for the detection of California (CAL) serogroup viruses and Cache Valley virus (CVV), for use in human surveillance. The targets selected...Full Text Available

2009-10-01

95

Construction and Analytical Application of Internal Amplification Controls (IAC) for Detection of Food Supply Chain-Relevant Viruses by Real-Time PCR-Based Assays

Invasive Aspergillosis

2011-09-07

96

British Library Electronic Table of Contents (United Kingdom)

Internal amplification controls (IACs) were constructed for incorporation into real-time nucleic acid amplification assays for bovine polyomavirus, hepatitis A virus, hepatitis E virus, human adenovirus, human norovirus genogroup I, human norovirus genogroup II, murine norovirus and porcine adenovirus. The addition of optimised amounts of IAC into the assays did not affect the limits of detection for each specific target virus. A poorly performed extraction of viral nucleic acids was simulated, and the effectiveness of IACs in identifying failed assays was demonstrated. The IACs constructed in this study can be reliably used in their specific assays to provide a robust control that can be routinely applied in the analysis of foods for viruses.

2011-01-01

97

Identification and characterization of retinoid-active short-chain dehydrogenases/reductases in Drosophila melanogaster

British Library Electronic Table of Contents (United Kingdom)

Background In chordates, retinoid metabolism is an important target of short-chain dehydrogenases/reductases (SDRs). It is not known whether SDRs play a role in retinoid metabolism of protostomes, such as Drosophila melanogaster. Methods Drosophila genome was searched for genes encoding proteins with ?50% identity to human retinol dehydrogenase 12 (RDH12). The corresponding proteins were expressed in Sf9 cells and biochemically characterized. Their phylogenetic relationships were analyzed using PHYLIP software. Results A total of six Drosophila SDR genes were identified. Five of these genes are clustered on chromosome 2 and one is located on chromosome X. The deduced proteins are 300 to 406 amino acids long and are associated with microsomal membranes. They recognize all-trans-retinaldehyd...

2009-01-01

98

A comparison of hepatic in vitro metabolism of T-2 toxin in rats, pigs, chickens, and carp

British Library Electronic Table of Contents (United Kingdom)

T-2 toxin, a highly toxic member of the type-A trichothecenes, is produced by various Fusarium moulds that can potentially affect human health. It is strongly cytotoxic for human hematopoietic progenitors. Alimentary toxic aleukia (ATA), a disease typically associated with human, is primarily induced by T-2 toxin. A comparison of the metabolism of T-2 toxin incubated with hepatocytes of rats, piglets, chickens, and the hepatic subcellular fractions (microsomes and cytosol) of piglets, chickens, rats, and carp (common carp and grass carp) was carried out. The activities of the recombinant pig CYP3A29 on the transformation of T-2 and HT-2 toxins were preliminary studied. Metabolites were identified by novel LC/MS-IT-TOF. Qualitative similarities and differences across the species were observ...

2011-01-01

99

Use of Organic Acids to Inactivate-Escherichia coli-O157:H7,-Salmonella-Typhimurium, and-Listeria monocytogenes-on Organic Fresh Apples and Lettuce

British Library Electronic Table of Contents (United Kingdom)

Abstract:- This study was undertaken to investigate the antimicrobial effect of organic acids against-Escherichia coli-O157:H7,-Salmonella-Typhimurium, and-Listeria monocytogenes-on whole red organic apples and lettuce. Several studies have been conducted to evaluate organic acids as sanitizers. However, no studies have compared antimicrobial effects of various organic acids on organic fresh produce, including evaluation of color changes of produce. Apples and lettuce were inoculated with a cocktail of 3 strains each of 3 foodborne pathogens provided above and treated with 1% and 2% organic acids (propionic, acetic, lactic, malic, and citric acid) for 0, 0.5, 1, 5, and 10 min. With increasing treatment time and acid concentration, organic acid treatments showed significant reduction compar...

2011-01-01

100

DOE assay methods used for characterization of contact-handled transuranic waste

DOE assay methods used for characterization of contact-handled transuranic waste

US Department of Energy methods used for characterization of contact-handled transuranic (CH-TRU) waste prior to shipment to the Waste Isolation Pilot Plant (WIPP) are described and listed by contractor site. The methods described are part of the certification process. All CH-TRU waste must be assayed for determination of fissile material content and decay heat values prior to shipment and prior to storage on-site. Both nondestructive assay (NDA) and destructive assay methods are discussed, and new NDA developments such as passive-action neutron (PAN) crate counter improvements and neutron imaging are detailed. Specifically addressed are assay method physics; applicability to CH-TRU wastes; calibration standards and implementation; operator training requirements and practices; assay procedures; assay precision, bias, and limit of detection; and ...

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101

Gamma irradiation in the control of pathogenic bacteria in refrigerated ground chicken meat; Irradiacao gama no controle de bacterias patogenicas em carne de frango refrigerada

US Department of Energy methods used for characterization of contact-handled transuranic (CH-TRU) waste prior to shipment to the Waste Isolation Pilot Plant (WIPP) are described and listed by contractor site. The methods described are part of the certification process. All CH-TRU waste must be assayed for determination of fissile material content and decay heat values prior to shipment and prior to storage on-site. Both nondestructive assay (NDA) and destructive assay methods are discussed, and new NDA developments such as passive-action neutron (PAN) crate counter improvements and neutron imaging are detailed. Specifically addressed are assay method physics; applicability to CH-TRU wastes; calibration standards and implementation; operator training requirements and practices; assay procedures; assay precision, bias, and limit of detection; and ...

1991-08-01

102

Toxicity of ultraviolet-irradiated halothane in mice

This work evaluated the effect of gamma radiation on reducing the population of Staphylococcus aureus, Escherichia coli nd Salmonella typhimurium in ground chicken breast stored under refrigeration. The experiment included a control and 4 doses of gamma radiation ( 2.0, 4.0, 6.0 and 8.0 kGy) along with 5 periods of storage under refrigeration ( 1, 7 14, 21 and 28 days). Samples of ground chicken breast were inoculated with Staphylococcus aureus (ATCC 14458), Escherichia coli (ATCC 11105) and Salmonella typhimurium (ATCC 0626), irradiated at temperatures between 4 and 8 deg C and stored under refrigeration (5 deg C) for 28 days. The increased radiation dose and period of storage under refrigeration caused a reduction of Staphylococcus aureus, Escherichia coli and Salmonella typhimurium populations in the ground chicken breast. Mean radiation D values determined for Staphylococcus aureus and Escherichia coli were 0.41 and ...

2000-09-01

103

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

One such agent is the widely used anesthetic, halothane. To study the toxicity of u.v. decomposed halothane, mice were exposed to anesthetic concentrations (1.3%) of non- and u.v.-irradiated halothane in oxygen for 90 min. Halothane sleeping times increased from 14.3 min to 72.5 min. Microsomal mixed function oxidase activity decreased, as shown by prolonged pentobarbital sleeping times 1 day after exposure to halothane and irradiated halothane (54.6 min and 149.1 min, respectively, as compared to a 34.6-min control). Quantitative and qualitative differences were found in the amount of (/sup 14/C)-pentobarbital metabolites excreted by u.v. irradiated halothane-exposed mice compared to either oxygen or non-irradiated halothane-exposed groups. In addition, serum glutamic-oxalacetic transaminase (SGOT) of irradiated halothane-exposed mice increased to 233% of the control values, and serum glutamic-pyruvic transaminase (SGPT) were 377% of control values. No significant ...

1982-01-01

104

Clofibrate-induced cytochrome P450-lauric acid omega hydroxylase(P450LA omega):purification, cDNA cloning, sequence and regulation

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

1991-04-01

105

Use of gold and silver standards based on phenol-formalde-hyde resin in assay-activation analysis of geological samples

A cytochrome P450 that hydroxylates lauric acid at the 12 position (P450LA omega) was isolated from liver microsomes of clofibrate treated rats. P450LA omega was immunologically distinct from P450s a,b,c,d,e,f,g,h,j,PB1, and PCN1. Polyclonal antibody against P450LA omega was utilized to screen a gt11 cDNA library. A clone (pP450LA omega), was isolated and its sequence determined. The P450LA omega mRNA is a minimum 2387 nts in length and codes for a P450 of Mr.58,222 daltons. This protein shares less than 35% amino acid similarity with P450s b,c,d,e,f,PB1, and PCN1; however, it does contain a hydrophobic amino terminal peptide and a conserved sequence surrounding the Cys residue at position 456, which is similar to other microsomal P450s. P450LA omega is present at high levels in untreated rat kidney and is induced by clofibrate in both kidney and liver. This induction is the result of an accumulation of mRNA through a rapid transcriptional ...

1986-05-01

106

A new simple multi-well plate-based assay for monocyte differentiation using human monocytic leukemia THP-1 cells.

Using standards on phenol-formaldehyde resin base for assaying-activation analysis of geological specimens for gold and silver has bee the advantage of uniformly distributing Au and Ag in spesimens and possible preparing tablets of practically any form or size. The validity and accuracy of these standards have been studied for the cases of short irradiation. Conventional point standards were used as reference standards. The experiments carried out have shown that tablet resol standards are suitable for a mass assaying-activation analysis for gold and silver at practically any concentrations.

107

A new simple multi-well plate-based assay for monocyte differentiation using human monocytic leukemia THP-1 cells

A new simple multi-well plate-based assay to determine monocyte differentiation-inducing activity was developed. It is based on the increase of macrophage adherence after the induction of monocyte differentiation. The assay was conducted using a human monocytic leukemia cell line, THP-1, treated with either 12-O-tetradecanoylphorbol 13-acetate or cytokines, such as tumor necrosis factor-alpha and interferon-gamma. This method is sensitive and easy to perform, especially in case of analyzing a large number of samples. PMID:17920620

2007-09-29

108

British Library Electronic Table of Contents (United Kingdom)

A new simple multi-well plate-based assay to determine monocyte differentiation-inducing activity was developed. It is based on the increase of macrophage adherence after the induction of monocyte differentiation. The assay was conducted using a human monocytic leukemia cell line, THP-1, treated with either 12-O-tetradecanoylphorbol 13-acetate or cytokines, such as tumor necrosis factor-? and interferon-?. This method is sensitive and easy to perform, especially in case of analyzing a large number of samples.

2007-01-01

109

Ovine reference materials and assays for prion genetic testing

Microscopic-Observation Drug-Susceptibility Assay for the Diagnosis of TB

BackgroundGenetic predisposition to scrapie in sheep is associated with several variations in the peptide sequence of the prion protein gene (PRNP). DNA-based tests...Full Text Available

110

Methanogenic Inhibition by Arsenic Compounds

BACKGROUNDNew diagnostic tools are urgently needed to interrupt the transmission of tuberculosis and multidrug-resistant tuberculosis. Rapid,...Full Text Available

2006-10-12

111

In Vitro Toxicity of Silver Nanoparticles in Human Lung ...

The acute acetoclastic methanogenic inhibition of several inorganic and organic arsenicals was assayed. Trivalent species, i.e., methylarsonous acid and arsenite, were highly inhibitory, with 50% inhibitory...Full Text Available

2004-09-01

112

Evaluating concentration estimation errors in ELISA microarray experiments

... The MTT assay indicates cell viability through a yellow to purple ... besiegers were said to have catapulted plague infected corpses into the city of ...

2009-03-01

113

Chemical Defense and the Persistence of Pioneer Plant Seeds in the Soil of a Tropical Cloud Forest:

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

114

British Library Electronic Table of Contents (United Kingdom)

ABSTRACT We present evidence that differences in soil seedbank persistence among pioneer plants in the cloud forest of Monteverde, Costa Rica, are influenced by differences in seed chemical defense. We used extracted seed chemicals from Bocconia frutescens (Papaveraceae), Guettarda poasana (Rubiaceae), Phytolacca rivinoides (Phytolaccaceae), Urera elata (Urticaceae), Cecropia polyphlebia (Cecropiaceae), and Witheringia meiantha (Solanaceae) to assess seed chemical defense in two ways: (1) a plant pathogen inhibition assay using Pythium irregulare; and (2) a brine shrimp toxicity assay using Artemia salina. The combined performance of each species in the two assays positively correlated with seedbank persistence. In the pathogen assay, mycelium growth was reduced when Pythium was cultured o...

2007-01-01

115

Antioxidant properties of 4-methylcoumarins in in vitro cell-free systems

British Library Electronic Table of Contents (United Kingdom)

4-Methylcoumarins that possess two hydroxyl groups ortho to each other in the benzenoid ring have shown to have excellent antioxidant and radical-scavenging properties in different experimental models. Furthermore, they cannot be metabolized by the liver P450 monoxygenases and thus cannot form 3,4-coumarin epoxides, which are believed to be mutagenic. Herein, we present a study on the structure activity relationship of eight synthetic 4-methylcoumarins, carried out by employing a series of different chemical cell-free tests. These compounds were tested by means of three assays involving one redox reaction with the oxidant (DPPH assay, ABTS+ assay and FRAP). Other assays were employed to evaluate the antioxidant properties of the coumarins under investigation against NO, O2- and HClO, which...

2010-01-01

116

A study of the incubation times of a new solid phase system for the assay of specific IgE with RAST

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

(1985). Italy Pisati, P. Aprile, C. Fondazione Clinica del Lavoro, Pavia (Italy).

117

Simultaneous Detection of Multiplex-Amplified Human Immunodeficiency Virus Type 1 RNA, Hepatitis C Virus RNA, and Hepatitis B Virus DNA Using a Flow Cytometer Microsphere-Based Hybridization Assay

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

2011-03-03

118

Rapid and Inexpensive Detection of Multidrug-Resistant Mycobacterium tuberculosis with the Nitrate Reductase Assay Using Liquid Medium and Direct Application to Sputum Samples?

The feasibility of performing a multiplex assay for the detection of human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV) RNAs and hepatitis B virus (HBV) DNA is demonstrated. This...Full Text Available

2000-03-01

119

Kidney-specific allo- and autoantibodies in the alloantibody response to rat kidney: the use of kidney homogenate as a target for serological analysis.

For rapid and low-cost detection of multidrug-resistant (MDR) Mycobacterium tuberculosis, we applied the nitrate reductase assay (NRA) using a liquid medium directly to sputum samples....Full Text Available

2008-10-01

120

False-Positive Results in a Recombinant Severe Acute Respiratory Syndrome-Associated Coronavirus (SARS-CoV) Nucleocapsid-Based Western Blot Assay Were Rectified by the Use of Two Subunits (S1 and S2) of Spike for Detection of Antibody to SARS-CoV

LEW anti-DA kidney and DA anti-LEW kidney sera were assayed using an indirect 125I anti-immunoglobulin-binding assay with kidney homogenate as target. This allowed the full spectrum of antibodies to...Full Text Available

1980-04-01

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121

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

To evaluate the reactivity of the recombinant proteins expressed in Escherichia coli strain BL21(DE3), a Western blot assay was performed by using a panel of 78 serum samples obtained,...Full Text Available

2006-03-01

122

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

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

1989-08-01

123

Comparison of Human Serum Dopamine-?-Hydroxylase Levels by Radioimmunoassay and Enzymatic Assay

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

1993-03-01

124

Protein binding assay for hyaluronate

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

1974-03-01

125

Comparison of in vitro disc diffusion and time kill-kinetic assays for the evaluation of antimicrobial wound dressing efficacy.

A relatively quick and simple assay for hyaluronate was developed using the specific binding protein, hyaluronectin. The hyaluronectin was obtained by homogenizing the brains of Sprague-Dawley rats, and then centrifuging the homogenate. The resulting supernatant was used as a source of crude hyaluronectin. In the binding assay, the hyaluronectin was mixed with (/sup 3/H)hyaluronate, followed by an equal volume of saturated (NH/sub 4/)/sub 2/SO/sub 4/, which precipitated the hyaluronectin and any (/sup 3/H)hyaluronate associated with it, but left free (/sup 3/H)hyaluronate in solution. The mixture was then centrifuged, and the amount of bound (/sup 3/H)hyaluronate in the precipitate was determined. Using this assay, the authors found that hyaluronectin specifically bound hyaluronate, since other glycosaminoglycans failed to compete for the binding protein. In addition, the interaction between hyaluronectin and hyaluronate ...

1986-11-01

126

A rapid, radiometric assay for sucrose synthase

There is a plethora of new silver-containing dressings on the market today. Various manufacturers attempt to show that their dressings are the most efficacious and therefore should be preferentially employed by health care workers based on the results of their in vitro tests. However, there have been no studies that clearly identify which tests are appropriate for comparison purposes. The purpose of this study was to determine which in vitro test is most appropriate for evaluating the antimicrobial efficacy of silver-containing dressings. This was done by testing seven silver-containing dressings and two non-silver-containing topical agents against 17 clinically relevant microorganisms using zone of inhibition assays and time-kill kinetic assays in complex media. The results for the two assays were then correlated to determine whether the methods generated similar results. It was determined that the two methods do not ...

127

Testing an agent-based model of bacterial cell motility: How nutrient concentration affects speed distribution

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 ...

1990-05-01

128

British Library Electronic Table of Contents (United Kingdom)

We revisit a recently proposed agent-based model of active biological motion and compare its predictions with own experimental findings for the speed distribution of bacterial cells, Salmonella typhimurium. Agents move according to a stochastic dynamics and use energy stored in an internal depot for metabolism and active motion. We discuss different assumptions of how the conversion from internal to kinetic energy d(v) may depend on the actual speed, to conclude that d 2 v ? with either ? = 2 or 1 ? < 2 are promising hypotheses. To test these, we compare the model?s prediction with the speed distribution of bacteria which were obtained in media of different nutrient concentration and at different times. We find that both hypotheses are in line with the experimental observations, with ? bet...

2011-01-01

129

Susceptibility to invasive bacterial infections in children with sickle cell disease

British Library Electronic Table of Contents (United Kingdom)

Individuals with sickle cell disease (SCD) demonstrate an increased susceptibility to invasive bacterial infections (IBI). The most common organisms causing IBI are Streptococcus pneumoniae, nontyphi Salmonella species and Haemophilus influenzae type b (Hib). IBI are the most common causes of death in children below 5 years of age with SCD. Increased susceptibility to IBI is because of several factors including dysfunctional antibody production and opsonophagocytosis as well as defective splenic clearance. Early diagnosis of Hib and pneumococcal infections combined with antibiotic prophylaxis and immunization programs, could lead to significant improvements in mortality, especially in Africa. Pediatr Blood Cancer. 2010;55:401-406. Copyright 2010 Wiley-Liss, Inc.

2010-01-01

130

Microbial pathogens in raw pork, chicken, and beef: benefit estimates for control using irradiation

Detection of irradiated chicken by ESR spectroscopy of bone

Various control procedures have been suggested for reducing foodborne infectious diseases. Receiving considerable attention is irradiation. This report estimates the medical and wage (or productivity) benefits associated with prevention of five human diseases transmitted by beef, pork, and chicken. (These diseases can also be transmitted by other vectors, such as eggs, milk, and pets. But these sources are not included in the analysis.) All of these foodborne infectious diseases - salmonellosis, campylobacteriosis, trichinosis, tapeworm, and toxoplasmosis - could be significantly reduced by irradiating meat and poultry. The Food and Drug Administration (FDA) has just approved irradiation of pork to prevent trichinosis (50FR 29658-59) and is considering approval of irradiation of chicken to kill Salmonella. 22 references.

1985-12-01

131

Identification of sucrose binding, membrane proteins using a photolyzable sucrose analog. [P. saccharophila

Ionizing radiation has been used to treat poultry to remove harmful microorganisms, mainly Salmonella, which contaminates chicken, goose and other fresh and frozen poultry. This microorganism is sensitive to low dose radiation. Thus, irradiating these foods with doses between 1 to 7 kGy results in a large reduction of bacteria. Since it is necessary to determine whether irradiation has occurred and to what extend, this work studied the signal produced by ionizing radiation within the hard crystalline matrix of chicken`s bone to establish a control method. Chicken`s drumsticks were irradiated and bones separated from flesh were lyophilized and milled. ESR spectrum was then obtained. The ESR signal increased linearly with dose over the range 0.25 to 8.0 kGy. Free radicals evaluated during 30 days after irradiation showed stable in this period. (Author).

1995-10-01

132

Evolution of nitrogen oxide(s) during in vivo nitrate reductase assay of soybean leaves

The sucrose derivative 6'-deoxy-6'-(2-hydroxy-4-azido)benzamidosucrose (6'-HABS) was prepared from sucrose (via 6'-deoxy-6'-aminosucrose) and 4-amino-salicylic acid. 6'-HABS is a competitive inhibitor of sucrose influx into protoplasts from developing soybean cotyledons and of sucrose binding to membranes from the bacteria P. saccharophila. The Ki for inhibition in the soybean protoplasts was 75..mu..M. 6'-Deoxy-6'-(2-hydroxy-3-/sup 125/Iodo-4-azido)benzamidosucrose was prepared by lactoperoxidase iodination of 6'-HABS. Upon photolysis in the presence of membranes from P saccharophila, label from the photoprobe is incorporated into a sucrose inducible polypeptide of mass 84 KD in SDS-PAGE. The polypeptide is protected from labeling by the inclusion of sucrose in the photolysis mixture. Photolysis conditions which lead to specific labeling of the sucrose protectable polypeptide in bacterial membranes also ...

1986-04-01

133

Development of an assay for a biomarker of pregnancy and early fetal loss

Studies were conducted to quantitate the evolution of nitrogen oxides (NO/sub (x)/) from soybean (Glycine max (L.) Merr.) leaves during in vivo nitrate reductase (NR) assays with aerobic and anaerobic gas purging. Anaerobic gas purging (N/sub 2/ and argon) consistently resulted in greater NO/sub (x)/ evolution than did aerobic gas purging (air and O/sub 2/). The evolution of NO/sub (x)/ was dependent on gas flow rate and on NO/sub 2//sup -/ formation in the assay medium; although a threshold level of NO/sub 2//sup -/ appeared to exist beyond which the rate of NO/sub (x)/ evolution did not increase further. The loss of NO/sub (x)/ from in vivo NR assays under gas purging explains partially, but not stoichiometrically, the decrease of NO/sub 2//sup -/ accumulation in in vivo NR assay medium with young soybean leaves. The lack of stoichiometry between NO/sub (x)/ evolution and apparent NO/sub 2//sup -/ ...

1981-12-01

134

Synthesis of novel tellurium containing analogues of choline and acetylcholine and their quantitation by pyrolysis-gas chromatography-mass spectrometry.

Human chorionic gonadotropin (hCG) is a glycoprotein hormone, secreted by the syncytiotrophoblast cells of the fertilized ovum, that enters the maternal circulation at the time of endometrial implantation. It is composed of two nonidentical subunits; ..cap alpha.. and ..beta.., with molecular weights of 14 kD and 23 kD, respectively. Human chorionic gonadotropin binds to the same receptor as hLH and displays the same biological response, namely, to stimulate the declining function of the corpus luteum to produce progestins and estrogen late in the menstrual cycle. The differences in the structures of hCG and hLH have been exploited to develop antibodies that can measure hCG specifically in the presence of hLH. Two-site antibody binding assays have been developed, based on a surface immunological concept of hCG epitopes, that involve four distinct regions to which antibodies against hCG can bind simultaneously. Antibody cooperative effects, in conjunction with ...

1987-10-01

135

Somatic mutations detected by immunofluorescence and flow cytometry

Methods for the synthesis and quantitation of the novel choline analogues, telluronium choline and acetyltelluronium choline, are described. An assay procedure utilizing pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) with cold trapping was developed with [2H4]telluronium choline and [2H4]acetyltelluronium choline as internal standards. The telluronium compounds were ion-pair extracted from tissue with dipicrylamine, washed with 2-butanone, and pyrolyzed prior to GC-MS analysis. The compounds were monitored using selected ion monitoring at m/z 232 and m/z 190 for acetyltelluronium and telluronium choline, respectively, and at m/z 236 and m/z 194 for the analogous deuterated internal standards. The assay was linear over a range of 20 pmol-20 nmol of compound taken through the assay. PMID:8130881

1993-12-31

136

Novel automated assay for the quality control of mexiletine hydrochloride formulations using sequential injection and on-line dilution

We have at hand the tools for development of both a single amino acid substitution and a gene loss type assay for somatic cell mutations in human red cells. We are close to being able to assess the assays as possible means for screening the human population for individuals who may be a risk of acquiring abnormally high frequencies of mutant cells. The cause for such a high frequency of mutant cells could be a prior exposure of the individual to large amount of mutagen, or it could indicate a member of a vulnerable subpopulation that may have poor DNA repair systems. Either of these cases could well be an indication that the particular individual is at high risk that exposure to mutagens could lead to potential pathology. In the near future we expect to determine the reliability of the assays and to perform experiments to confirm the validity of flow cytometry in measuring real somatic mutagenic events that occur in vivo. ...

137

British Library Electronic Table of Contents (United Kingdom)

The first automated method for the determination of mexiletine hydrochloride - an antiarrhythmic agent - is reported. The method is based on the reaction of the analyte with o-phthalaldehyde (OPA) in the presence of sulfite in basic medium using a sequential injection (SI) manifold. The reaction product was monitored spectrofluorimetrically (lex=350nm/lem=446nm). A simple and effective on-line dilution approach was adopted in order to expand the linearity and apply the method to assay, dosage uniformity and dissolution tests with minimum sample preparation. Chemical (pH, amount concentrations of OPA and sulfite) and instrumental variables (temperature, flow rate, injection volumes, etc.) that affected the determination were studied. The developed assay was validated in terms of linearity, ...

2008-01-01

138

Efficient methodologies for sensitive HIV-1 RNA quantitation from plasma and vaginal secretions

British Library Electronic Table of Contents (United Kingdom)

Background: Quantifying HIV levels in mucosal secretions is essential to study compartmentalized expression of HIV and facilitate development of intervention strategies to prevent disease progression and transmission. Objectives: To develop a sensitive, reliable, and cost-effective technique to quantify HIV from blood and vaginal secretions that is compatible with efficient implementation in clinical research environments. Study design: A sensitive, reliable, internally-controlled real-time reverse transcriptase (RT) PCR assay, which uses the HIV-1 pol gene as a target (Hpol assay) was developed to quantify HIV levels in plasma and genital secretions, and compared to the widely used Roche Amplicor(TM) HIV-1 Monitor assay. In addition, a simplified method of sample collection and processing...

2009-01-01

139

Correction factors determination in large samples gamma assay using its own multi-gamma lines spectrum

Correction factors determination in large samples gamma assay using its own multi-gamma lines spectrum

An easy and simple method for gamma assay of large multi-gamma lines samples was introduced in this work. This method performs the assay using point source calibration. The correction factors for volume and self-attenuation are experimentally deduced from the spectra of different thicknesses samples utilizing the following two simple well known facts: large and small samples of the same homogenous material have identical specific activities; the self-attenuation of gamma line decreases as its energy increases. The method was successfully applied to naturally occurring radioactive material (NORM) large samples. This method does not require complicated mathematical procedures. Neither sample matrix data nor detector unit composition is needed.

2009-10-15

140

Correction factors determination in large samples gamma assay using its own multi gamma lines spectrum

An easy and simple method for gamma assay of large multi-gamma lines samples was introduced in this work. This method performs the assay using point source calibration. The correction factors for volume and self-attenuation are experimentally deduced from the spectra of different thicknesses samples utilizing the following two simple well known facts: large and small samples of the same homogenous material have identical specific activities; the self-attenuation of gamma line decreases as its energy increases. The method was successfully applied to naturally occurring radioactive material (NORM) large samples. This method does not require complicated mathematical procedures. Neither sample matrix data nor detector unit composition is needed.

2009-10-01

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141

A mass action model of the dose-response curve of immunoradiometric assay and its curve fitting

An easy and simple method for gamma assay of large multi gamma lines samples was introduced in this work. This method performs the assay using point source calibration. The correction factors for volume and self-attenuation are experimentally deduced from the spectra of different thicknesses samples utilizing the following tow simple well known facts: Large and small samples of the same homogenous material have identical activities; the self attenuation of gamma line decreases as its energy increases. The method was successfully applied to NORM (Naturally Occurring Radioactive Material) large samples. This method doesn't require complicated mathematical procedures. Neither sample matrix data nor detector unit composition is needed. (author)

142

Murine antibody response to oral infection with live aroA recombinant Salmonella dublin vaccine strains expressing filamentous hemagglutinin antigen from Bordetella pertussis.

In view of no satisfactory mathematical model is presently available for the dose-response curve of immunoradiometric assay, a three parameter model based on single binding site mass action law has been derived. A curve fitting method based on similar principle of linear robust regression was designed and a software was prepared for use on IBM personal computers. Experiments revealed that the model is applicable to a variety of IRMA systems as well as to time-resolved immunofluorometric assay of hAFP. When there was outlier(s) of the calibration curve, the average bias of unknown samples obtained with this method is significantly smaller than methods using four parameter logistic model or four parameter single site mass action model.

143

Using a Pan-Viral Microarray Assay (Virochip) to Screen Clinical Samples for Viral Pathogens

Two plasmids which express either nearly intact or truncated filamentous hemagglutinin (FHA) from Bordetella pertussis and which are marked with a tetracycline resistance (Tcr) gene were transformed into Salmonella dublin SL1438, an aroA deletion mutant intended for use as an attenuated oral vaccine against salmonellosis. These S. dublin recombinants, when fed to mice, induced serum immunoglobulin, immunoglobulin M (IgM), and sometimes IgA antibody responses to FHA and S. dublin. In addition, IgA antibodies against FHA were found in gut wash fluids. S. dublin carrying pDB2300, a multicopy plasmid encoding truncated FHA protein, induced a better antibody response than did S. dublin carrying pDB2000, a low-copy-number plasmid encoding full-sized FHA. Administration of tetracycline to mice enhanced the stability of recombinant plasmids, and tetracycline-treated mice developed higher anti-FHA titers. Although neither strain examined is suitable for use in a human oral ...

1990-08-01

144

Uptake and Intracellular Activity of Moxifloxacin in Human Neutrophils and Tissue-Cultured Epithelial Cells

The diagnosis of viral causes of many infectious diseases is difficult due to the inherent sequence diversity of viruses as well as the ongoing emergence of novel viral pathogens, such as SARS coronavirus...Full Text Available

145

Transuranic contaminated waste form characterization and data base

The penetration by moxifloxacin of human neutrophils (polymorphonuclear leukocytes [PMN]) and tissue-cultured epithelial cells (McCoy cells) was evaluated by a fluorometric assay. At...Full Text Available

1999-01-01

146

Tissue distribution of brain-thymus shared antigens recognized by anti-brain xenosera in the rat, dog and man.

This volume contains 5 appendices. Title listing are: technologies for recovery of transuranics; nondestructive assay of TRU contaminated wastes; miscellaneous waste characteristics; acceptance criteria for TRU waste; and TRU waste treatment technologies.

1980-07-01

147

Staphylococcal Enterotoxin B: Solid-Phase Radioimmunoassay

A comparative and quantitative study of the tissue distribution of brain-thymus shared antigens was carried out using rabbit antisera to rat, dog and human brain homogenates, assayed on rat, dog and...Full Text Available

1979-03-01

148

Significance of arming, potentiating and blocking factors as correlates the tumour-host interaction in the hamster SV40 system.

An immunoassay employing ¹²⁵I-labeled enterotoxin B and polystyrene tubes coated with specific antibody was used for assaying purified and crude enterotoxin. Antibody was adsorbed to untreated...Full Text Available

1971-11-01

149

Serum HER2 Level Measured by Dot Blot: A Valid and Inexpensive Assay for Monitoring Breast Cancer Progression

The study of blocking factors requires in vitro assay of cell mediated immunity that parallels the in vivo response. By microcytotoxicity testing, progressor and immune peripheral blood lymphocytes...Full Text Available

1975-12-01

150

Seasonal Patterns of Nitrate Reductase and Nitrogenase Activities in Phaseolus vulgaris L. ¹

Human epidermal growth factor receptor 2 (HER2) is one of the most important prognostic and predictive factors for breast cancer patients. Recently, serum HER2...Full Text Available

151

Scintillation proximity assay for measurement of RNA methylation

The patterns of nitrate reductase activity (NRA) in the leaves (in vivo assay) and root nodule nitrogenase activity (C₂H₂ reduction) were investigated throughout...Full Text Available

1979-03-01

152

SWISS MADE: Standardized WithIn Class Sum of Squares to Evaluate Methodologies and Dataset Elements

Methylation of RNA by methyltransferases is a phylogenetically ubiquitous post-transcriptional modification that occurs most extensively in transfer RNA (tRNA) and ribosomal RNA (rRNA). Biochemical...Full Text Available

2009-03-01

153

Reduction of Nitrate and Nitrite in Lambsquarters (Chenopodium album) Biotypes Resistant and Susceptible to Atrazine Toxicity ¹

Contemporary high dimensional biological assays, such as mRNA expression microarrays, regularly involve multiple data processing steps, such as experimental processing, computational processing, sample...Full Text Available

154

Rapid and Inexpensive Drug Susceptibility Testing of Mycobacterium tuberculosis with a Nitrate Reductase Assay

The nitrite-reducing activity of the normal susceptible biotype of lambsquarters (Chenopodium album L.) was strongly inhibited by atrazine in the assay medium, both in the case of the...Full Text Available

1980-05-01

155

Rapid Detection of Ofloxacin Resistance in Mycobacterium tuberculosis by Two Low-Cost Colorimetric Methods: Resazurin and Nitrate Reductase Assays

Multidrug-resistant tuberculosis is an increasing public health concern in many parts of the world, especially in low-income countries, where most cases occur. Traditional drug susceptibility testing...Full Text Available

2002-02-01

156

Quantitative PCR Assay for Mycobacterium pseudoshottsii and Mycobacterium shottsii and Application to Environmental Samples and Fishes from the Chesapeake Bay?

We have evaluated the performance of two rapid, low-cost methods for the detection of ofloxacin (OFX) resistance with 95 Mycobacterium tuberculosis isolates from countries with high...Full Text Available

2005-04-01

157

Purification of mRNA guanylyltransferase from calf thymus.

Striped bass (Morone saxatilis) in the Chesapeake Bay are currently experiencing a very high prevalence of mycobacteriosis associated with newly described Mycobacterium...Full Text Available

2010-09-01

158

Production of placental alkaline phosphatase (PLAP) and PLAP-like material by epithelial germ cell and non-germ cell tumours in vitro.

mRNA guanylyltransferase has been extensively purified from calf thymus. A GTP-binding assay was used based on the observations by Shuman and Hurwitz (1981) and Venkatesan and Moss (1982) that vaccinia...Full Text Available

1982-01-01

159

Precise Detection and Tracing of Trichoderma hamatum 382 in Compost-Amended Potting Mixes by Using Molecular Markers

Placental and placental-like alkaline phosphatase (PLAP) levels in the culture media of 87 cell lines of neoplastic and 'normal' origin were measured by a conventional immunosorbent enzymatic assay...Full Text Available

1994-02-01

160

Observations on the use of solid-phase-coupled antibodies in the radioimmunoassay of human placental lactogen

Randomly amplified polymorphic DNA (RAPD) analysis and the PCR assay were used in combination with dilution plating on a semiselective medium to detect and enumerate propagules of Trichoderma...Full Text Available

1999-12-01

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

161

Nitric Oxide and Nitrous Oxide Production by Soybean and Winged Bean during the in Vivo Nitrate Reductase Assay ¹

A detailed comparative assessment was made of the use of solid-phase-coupled antibodies in radioimmunoassay, by using an assay for human placental lactogen as a model system. The major advantages of...Full Text Available

1974-03-01

162

Nitrate Reductase Activity in Soybeans (Glycine max [L.] Merr.)

This study was conducted to determine by gas chromatography (GC) and mass spectrometry (MS) the identity and the quantity of volatile N products produced during the helium-purged in vivo...Full Text Available

1986-11-01

163

Myosin Motors Drive Long Range Alignment of Actin Filaments*

The optimum in vivo nitrate reductase (NR) assay medium for soybean (Glycine max [L.] Merr.) leaves was 50 mm KNO₃, 1% (v/v) 1- propanol, and 100...Full Text Available

1976-12-01

164

Mutagenicity studies in a tyre plant: in vitro activity of workers' urinary concentrates and raw materials.

The bulk alignment of actin filament sliding movement, powered by randomly oriented myosin molecules, has been observed and studied using an in vitro motility assay. The well established,...Full Text Available

2010-02-12

165

Multiplexed, rapid detection of H5N1 using a PCR-free nanoparticle-based genomic microarray assay

The possible contribution to urinary mutagenicity of occupational exposures in the rubber industry was studied by assaying the urine concentrates of 72 workmen (44 smokers) employed in a tyre plant....Full Text Available

1985-07-01

166

Monitoring digoxin therapy: I. Plasma concentrations and an in vitro assay of tissue response.

BackgroundFor more than a decade there has been increasing interest in the use of nanotechnology and microarray platforms for diagnostic applications. In this report, we describe...Full Text Available

167

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry assay for organophosphorus toxicants bound to human albumin at Tyr411

1 An in vitro technique is described for measuring the uptake of 86Rb by human erythrocytes. 2 Fifteen patients were treated with digoxin for atrial fibrillation and other fast arrhythmias. 3 86Rb uptake...Full Text Available

1977-04-01

168

Lipotoxic heart disease in obese rats: Implications for human obesity

Our goal was to determine whether chlorpyrifos oxon, dichlorvos, diisopropylfluorophosphate (DFP), and sarin covalently bind to human albumin. Human albumin or plasma was treated with organophosphorus...Full Text Available

2007-02-15

169

Inhibition of beta-lactamase of Bacillus licheniformis 749/C by compound PS-5, a new beta-lactam antibiotic.

To determine the mechanism of the cardiac dilatation and reduced contractility of obese Zucker Diabetic Fatty rats, myocardial triacylglycerol (TG) was assayed chemically and morphologically. TG was...Full Text Available

2000-02-15

170

Induction of mortality and malformation in Xenopus laevis embryos by water sources associated with field frog deformities.

By use of a new computer-assisted u.v.-spectrophotometric assay method, the kinetic parameters of the reaction catalysed by Bacillus licheniformis 749/C beta-lactamase were re-examined and the mode...Full Text Available

1980-01-01

171

Increases in mouse uterine heat shock protein levels are a sensitive and specific response to uterotrophic agents.

Water samples from several ponds in Minnesota were evaluated for their capacity to induce malformations in embryos of Xenopus laevis. The FETAX assay was used to assess the occurrence of malformations...Full Text Available

1998-12-01

172

In vitro analysis of nuclear mRNA export using molecular beacons for target detection

There is increasing consensus that the uterotrophic estrogenicity assay should be coupled with other morphometric or molecular end points that might enhance its sensitivity. We have previously shown...Full Text Available

2002-12-01

173

In Vivo Measurement of Indole-3-acetic Acid Decarboxylation in Aging Coleus Petiole Sections

A detailed molecular characterization of nuclear mRNA export will require an in vitro system, allowing a biochemical reconstitution of transport. To this end, an mRNA export assay has...Full Text Available

2003-06-01

174

Immunoquantitative Real-Time PCR for Detection and Quantification of Staphylococcus aureus Enterotoxin B in Foods

The concentration of indoleacetic acid (IAA) in plant tissues is regulated, in part, by its rate of decarboxylation. However, the commonly used in vitro assays for IAA oxidase may not...Full Text Available

1983-09-01

175

Identification of environmental chemicals with estrogenic activity using a combination of in vitro assays.

A real-time immunoquantitative PCR (iqPCR) method for detection of Staphylococcus aureus enterotoxin B (SEB) was developed and evaluated using both pure cultures and foods. The assay...Full Text Available

2006-10-01

176

High-density multiplex detection of nucleic acid sequences: oligonucleotide ligation assay and sequence-coded separation.

Environmental chemicals that function as estrogens have been suggested to be associated with an increase in disease and dysfunctions in animals and humans. To characterize chemicals that may act as...Full Text Available

1996-10-01

177

Genetic relationship between anxiety- and fear -related behaviors in BXD recombinant inbred mice

We describe a non-isotopic, semi-automated method for large-scale multiplex analysis of nucleic acid sequences, using the cystic fibrosis transmembrane regulator (CFTR) gene as an example. Products...Full Text Available

1994-10-25

178

Genetic Diversity in Blastomyces dermatitidis: Implications for PCR Detection in Clinical and Environmental Samples

Mood and anxiety disorders and rodent phenotypic measures modeling these disorders have a strong genetic component. Various assays are used to study the neurobiological basis of fear- and anxiety-related...Full Text Available

2009-03-01

179

Field application of a colorimetric method of assaying chloroquine and desethylchloroquine in urine

SummaryBlastomycosis is a serious and potentially fatal infection by the thermally dimorphic fungus Blastomyces dermatitidis. PCR assays targeting the BAD-1 virulence...Full Text Available

2010-03-01

180

Factors affecting actions of ethanol on GABA-activated chloride channels

In a study in western Kenya of malaria-infected adult women who had been treated with chloroquine, we compared the level of chloroquine and its principal metabolite, desethylchloroquine, in urine,...Full Text Available

1988-01-01

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181

Evolution of Nitrogen Oxide(s) during In Vivo Nitrate Reductase Assay of Soybean Leaves

Effects of ethanol in vitro on membrane vesicles were evaluated by monitoring "3"6Cl"- influx. Different assay parameters were tested to determine increased or decreased action of ethanol on GABA-activated chloride channels. The ability of 30 mM ethanol to augment "3"6Cl"- flux was seen at 0 degree C, in the absence of GABA, and at 34 degree C in the presence of GABA, using two different assay procedures. Picrotoxin blocked the direct effects of ethanol suggesting GABA_a involvement. Endogenous GABA in the medium surrounding the microsacs was assayed at different temperatures both in the presence and absence of GABA and ethanol. The direct effect of ethanol did not appear to involve the action of endogenous GABA. In addition to temperature effects on the assay, time of membrane storage also influenced ethanol action. Microsacs stored on ice for 2 hours or more lost their ability to respond to ethanol ...

182

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

Studies were conducted to quantitate the evolution of nitrogen oxides (NO_(x)) from soybean [Glycine max (L.) Merr.] leaves during in vivo nitrate reductase...Full Text Available

1981-12-01

183

Efficient discovery of ASCL1 regulatory sequences through transgene pooling

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

2003-02-01

184

Dynamic SpoIIIE assembly mediates septal membrane fission during Bacillus subtilis sporulation

Zebrafish transgenesis is a powerful and increasingly common strategy to assay vertebrate transcriptional regulatory control. Several challenges remain, however, to the broader application of...Full Text Available

2010-06-01

185

Drug Susceptibility Testing of Mycobacterium tuberculosis by a Nitrate Reductase Assay Applied Directly on Microscopy-Positive Sputum Samples

SpoIIIE is an FtsK-related protein that transports the forespore chromosome across the Bacillus subtilis sporulation septum. We use membrane photobleaching and protoplast assays to...Full Text Available

2010-06-01

186

Differential Effect of Irradiance and Nutrient Nitrate on the Relationship of in Vivo and in Vitro Nitrate Reductase Assay in Chlorophyllous Tissues ¹

Current methods for drug susceptibility testing of Mycobacterium tuberculosis are either costly or slow. As the prevalence of multidrug-resistant strains increases, the need for fast,...Full Text Available

2005-07-01

187

Development and Application of Pathovar-Specific Monoclonal Antibodies That Recognize the Lipopolysaccharide O Antigen and the Type IV Fimbriae of Xanthomonas hyacinthi

Growth at increasing continuous irradiance (at high nutrient nitrate) and nutrient nitrate concentrations (at high continuous irradiance) furnished increases in the in vivo and in...Full Text Available

1977-04-01

188

Determination of Salirasib (S-trans, trans-farnesylthiosalicylic acid) in human plasma using liquid chromatography-tandem mass spectrometry

The objective of this study was to develop a specific immunological diagnostic assay for yellow disease in hyacinths, using monoclonal antibodies (MAbs). Mice were immunized with a crude cell wall preparation...Full Text Available

1999-09-01

189

Detection of Ockelbo virus RNA in skin biopsies by polymerase chain reaction.

A liquid chromatography/tandem mass spectrometric (LC/MS/MS) assay was developed for the quantitative determination of salirasib (S-trans, trans-farnesylthiosalicylic acid,...Full Text Available

2008-06-15

190

Detection and Toxin Typing of Clostridium perfringens in Formalin-Fixed, Paraffin-Embedded Tissue Samples by PCR?

A sensitive assay based on the polymerase chain reaction for the detection of Ockelbo virus RNA was developed. Two primer pairs from the gene coding for the E2 glycoprotein were chosen. By use of a...Full Text Available

1993-08-01

191

Comparison of ante-mortem assays to assess progression/regression of paratuberculosis in individual dairy animals

Since current microbiology methods are not suitable to detect Clostridium perfringens in formalin-fixed, paraffin-embedded tissue samples, we developed a PCR assay to detect toxin-encoding...Full Text Available

2009-03-01

192

Comparative Inhibitory Effects of Antigen and Antibody in the Staphylococcal Enterotoxin Solid-Phase Radioimmunoassay System

Johne disease, caused by Mycobacterium avium, subspecies paratuberculosis (MAP) is becoming increasingly widespread on dairy farms worldwide, due in part, to the absence...Full Text Available

2010-05-01

193

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

A solid-phase radioimmunoassay employing ¹²⁵I-labeled enterotoxins and polystyrene tubes coated with specific antibody has been developed for assaying the relative concentrations of antibodies...Full Text Available

1975-06-01

194

Association of elevated levels of extracellular neuraminidase with clinical isolates of type III group B streptococci.

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

1987-01-01

195

Applications and Experience with PCR-Based Assays to Predict Blood Group Antigens

The level of total extracellular neuraminidase produced by 74 clinical isolates of group B streptococci isolated from diseased or asymptomatically colonized infants was assayed. Extracellular neuraminidase...Full Text Available

1978-09-01

196

Application of tumor, bacterial and parasite susceptibility assays to study immune alterations induced by environmental chemicals

SummaryDNA-based tests are increasingly being used to predict a blood group phenotype. This is possible because genes encoding 29 of the 30 blood group systems have been cloned and sequenced,...Full Text Available

2009-01-01

197

Antibody avidity in swine lymphocyte antigen-defined miniature pigs.

Model systems to study the effects of chemicals of environmental concern on bacterial and parasitic diseases as well as the immunosurveillance and destruction of transplantable tumor cells were described...Full Text Available

1982-02-01

198

Anti-beta-endorphin immunoglobulin G in humans.

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

1992-10-01

199

Anion-exchange extraction of cephapirin, cefotaxime, and cefoxitin from serum for liquid chromatography.

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

1986-11-01

200

An improved method for undertaking limiting dilution assays for in vitro cloning of Plasmodium falciparum parasites

An anion-exchange column technique for extraction of antibiotics from serum proteins has been developed for use in the assay of cephapirin, cefotaxime, and cefoxitin by high-pressure liquid chromatography....Full Text Available

1982-04-01

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

201

Adaptations of the Saker-Solomons test: simple, reliable colorimetric field assays for chloroquine and its metabolites in urine.

BackgroundObtaining single parasite clones is required for many techniques in malaria research. Cloning by limiting dilution using microscopy-based assessment for parasite growth...Full Text Available

202

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

Two field-adapted colorimetric methods for measuring the antimalarial drug chloroquine in urine are described. Both are modifications of the method of Saker and Solomons for screening urine for phencyclidine...Full Text Available

1989-01-01

203

A simplified assay for the arylamine N-acetyltransferase 2 polymorphism validated by phenotyping with isoniazid.

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

204

A non-radioactive dot-blot assay for transglutaminase activity

Human arylamine N-acetyltransferase (NAT) activity is determined by two distinct genes, NAT1 and NAT2, and the classical acetylation polymorphism in NAT2 has been associated with a variety of disorders,...Full Text Available

1997-09-01

205

A double antibody solid phase assay for DNA autoantibodies for clinical use.

Aberrant transglutaminase (TG) activity has been implicated in the pathology of numerous diseases including Huntington disease and Alzheimer disease. To fully characterize the role of TGs in...Full Text Available

2009-07-01

206

A convenient and sensitive allergy test: IgE crosslinking-induced luciferase expression in cultured mast cells

DNA antoantibodies in serum will bind to antigen-coated polystyrene tubes and can be detected by radiolabelled anti-immunoglobulin. The method is quantitative, gives information on the antibody class...Full Text Available

1976-08-01

207

A Simple, Versatile and Sensitive Cell-Based Assay for Prions from Various Species

BackgroundFor the detection of allergen-specific IgE in sera, solid-phase IgE-binding assays like the CAP test are commonly used. Although such immunochemical methods are very sensitive,...Full Text Available

2010-10-01

208

A Simple Colorimetric Assay for Specific Detection of Glutathione-S Transferase Activity Associated with DDT Resistance in Mosquitoes

Detection and quantification of prion infectivity is a crucial step for various fundamental and applied aspects of prion research. Identification of cell lines highly sensitive to prion infection led...Full Text Available

209

A Novel Information Retrieval Model for High-Throughput Molecular Medicine Modalities

BackgroundInsecticide-based methods represent the most effective means of blocking the transmission of vector borne diseases. However, insecticide resistance poses a serious threat...Full Text Available

210

Post-transplant monitoring of renal allografts: are we there yet?

Significant research has been devoted to predicting diagnosis, prognosis, and response to treatment using high-throughput assays. Rapid translation into clinical results hinges upon efficient access...Full Text Available

211

British Library Electronic Table of Contents (United Kingdom)

Transplantation has emerged as the therapy of choice for many patients with end organ failure. One of the major goals is to tailor immunosuppressive therapy to the individual needs of every patient to balance the risk for rejection and over-immunosuppression. This will require diagnostic tools that can detect harmful processes in the allograft early, and that can be measured repeatedly. This review will consider recent advances in our understanding of the molecular nature of these processes and how this information is being utilized to design novel diagnostic assays to non-invasively monitor allografts. Highlighted is the need for large-scale prospective multi-centre studies to validate assays that show early promise in single centre studies.

2009-01-01

212

Evaluation of the Performance of the Automated NucliSENS easyMAG and EasyQ Systems versus the Roche AmpliPrep-AMPLICOR Combination for High-Throughput Monitoring of Human Immunodeficiency Virus Load?

An integrated Tomographic Gamma Scanning system for non-destructive assay of radioactive waste

This study presents the data of an evaluation of the automated Nuclisens easyMAG and EasyQ systems versus the Roche AmpliPrep-AMPLICOR combination for testing of high-volume human immunodeficiency virus (HIV) load. This represents a follow-up of a previous study investigating the performance of the real-time Nuclisens assay using the semiautomated NucliSENS miniMAG extraction procedure. Three hundred eighteen patient samples were analyzed using both methods. The easyMAG-EasyQ HIV type 1 system has a higher sensitivity and broader dynamic range than the Cobas AmpliPrep-AMPLICOR system when the standard Roche assay is used alone, 25 to 3,000,000 IU/ml versus 400 to 750,000 HIV RNA copies/ml, respectively. There was significant correlation between the assays (0.93; P < 0.0001), with good accuracy (percent similarity mean ? = 96%), good precision (percent similarity standard deviation = 4.97%), and overall good agreement ...

2007-05-01

213

An integrated Tomographic Gamma Scanning system for non-destructive assay of radioactive waste

The Tomographic Gamma Scanning (TGS) technique is a relatively new method in the field of non-destructive assay (NDA) of radioactive waste. The TGS technique combines High Resolution Gamma Spectrometry (HRGS) with Three-Dimensional (3-D) low spatial resolution transmission and emission image reconstruction techniques to achieve assay goals. When compared to the traditional methods such as Segmented Gamma Scanning (SGS), the TGS technique can yield better accuracies for cases where the radionuclide is distributed non-uniformly in a heterogeneous matrix. The TGS technique is ideally suited for low-to-moderate density waste matrices, say 1.0 g cm{sup -3} or below for 55 US gal. drums, although it can be extended to higher densities by using alternative approaches to the design or analyses. Recently Canberra Industries designed, built and characterized four such TGS systems for nuclear power plant applications. Many of the design features and the ...

2007-08-21

214

USE OF A CONTROLLED CHLORINE DIOXIDE (CLO2) RELEASE SYSTEM IN COMBINATION WITH MODIFIED ATMOSPHERE PACKAGING (MAP) TO CONTROL THE GROWTH OF PATHOGENS

The Tomographic Gamma Scanning (TGS) technique is a relatively new method in the field of non-destructive assay (NDA) of radioactive waste. The TGS technique combines High Resolution Gamma Spectrometry (HRGS) with Three-Dimensional (3-D) low spatial resolution transmission and emission image reconstruction techniques to achieve assay goals. When compared to the traditional methods such as Segmented Gamma Scanning (SGS), the TGS technique can yield better accuracies for cases where the radionuclide is distributed non-uniformly in a heterogeneous matrix. The TGS technique is ideally suited for low-to-moderate density waste matrices, say 1.0 g cm"-"3 or below for 55 US gal. drums, although it can be extended to higher densities by using alternative approaches to the design or analyses. Recently Canberra Industries designed, built and characterized four such TGS systems for nuclear power plant applications. Many of the design features and the end ...

2007-08-21

215

British Library Electronic Table of Contents (United Kingdom)

Abstract Salmonella Typhimurium and Listeria monocytogenes are major bacterial pathogens associated with poultry products. A controlled released ClO2 sachet applied with modified atmosphere packaging (MAP) was evaluated for its ability to control the growth of S. Typhimurium and L. monocytogenes on raw chicken breast during refrigerated storage. The fresh chicken samples were inoculated with one or the other of the pathogens at 104-cfu/g, and the packages (with and without ClO2 sachet) were flushed with ambient air or 30% CO2/70% N2 before sealing, and then stored at 4C for up to 21 days. The maximum reduction in MAP plus ClO2 (compared with MAP alone) was 0.68-log-cfu/g for S. Typhimurium and 1.87-log-cfu/g for L. monocytogenes. Color and pH changes of the chicken breast were observed at ...

2011-01-01

216

Antioxidant and toxicity tests of roasted noni (Morinda citrifolia) leaf infusion

British Library Electronic Table of Contents (United Kingdom)

Summary The antioxidant properties and toxicity profile of roasted noni (Morinda citrifolia L.) leaf infusion were evaluated. The 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging activity was greater than green tea infusion (81.6 +- 0.9% vs. 57.5 +- 1.8%, P < 0.001). The mean quercetin and kaempferol contents of roasted noni leaf infusion, as prepared by the consumer, were 0.24 +- 0.01 and 0.14 +- 0.01 mg mL-1, respectively. Tannic acid content was 10 +- 1 mg mL-1. The infusion was non-mutagenic in the reverse mutation test in Salmonella typhimurium and did not induce primary DNA damage in E. coli PQ37. Further, no significant primary DNA damage was induced by 5,15-dimethylmorindol, which was the only detectable anthraquinone in noni leaves. The infusion was not cytotoxic in the 24 h b...

2009-01-01

217

Active packaging of ground beef patties by edible zein films incorporated with partially purified lysozyme and Na2EDTA

British Library Electronic Table of Contents (United Kingdom)

Summary In this study, antimicrobial activity of zein films incorporated with partially purified lysozyme and disodium ethylenediaminetetraacetic acid (Na2EDTA) has been tested on selected pathogenic bacteria and refrigerated ground beef patties. The developed films containing 700-g-cm-2 lysozyme and 300-g-cm-2 Na2EDTA showed antimicrobial activity on Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella typhimurium. The application of lysozyme and Na2EDTA incorporated zein films on beef patties significantly decreased total viable counts (TVC) and total coliform counts after 5-days of storage compared to those of control patties (P-2EDTA or Na2EDTA alone significantly slowed down the oxidative changes in patties during storage (P-P-2EDTA for active packaging of refrigerated mea...

2011-01-01

218

Use of receptor affinity chromatography in purification of the growth hormone-like factor produced by plerocercoids of the tapeworm Spirometra mansonoides.

Radiation inactivation target size of rat adipocyte glucose transporter

The plerocercoid stage of the tapeworm Spirometra mansonoides produces a functional analog of human growth hormone (hGH). Among the similarities between plerocercoid growth factor (PGF) and hGH is competition for the same receptors on rabbit liver membranes. To take advantage of this characteristic in a purification scheme for PGF, rabbit liver microsomes were solubilized in Triton X-100 and the hGH receptors were purified over an hGH affinity column. The purified receptors from six rabbit livers were coupled to Affi-Gel-10 to create a receptor affinity column which was used to purify PGF. Chromatography of crude PGF over the receptor column resulted in a 1044 fold increase in specific activity. SDS-PAGE in the presence of 2-mercaptoethanol showed that the affinity-purified PGF contained three protein bands with apparent Mrs of 27.5 K, 22 K, and 16.7 K. Injections of the partially-purified PGF into hypophysectomized rats produced a dose-dependent growth response ...

1988-01-01

219

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

In situ assembly states of rat adipocyte glucose transport protein in plasma membrane (PM) and in microsomal pool (MM) were assessed by measuring target size (TS) of D glucose-sensitive, cytochalasin B binding activity. High energy radiation inactivated the binding in both PM and MM by reducing the total capacity of the binding (B/sub T/) without affecting the dissociation constant (K/sub D/). The reduction in B/sub T/ as a function of radiation dose was analyzed based on classical target theory, from which TS was calculated. TS in the PM of insulin-treated adipocytes was 58 KDa. TS in the MM of noninsulin-treated and insulin-treated adipocytes were 112 and 109 KDa, respectively. With MM, however, inactivation data showed anomalously low radiation sensitivities at low radiation doses showing a shoulder in the semilog plots, which may be due to an interaction with a radiation sensitive inhibitor. With these results, they propose the following model: Adipocyte ...

1987-05-01

220

Induction of lauric acid omega-hydroxylation by peroxisomal proliferators in bluegill and catfish

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 exogenous substrates. These data suggest that the ...

1986-06-15

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221

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

Peroxisome proliferating agents (PPAs) are a structurally diverse group of chemicals that include environmental chemical contaminants such as certain chlorinated herbicides, solvents and plasticizers. PPAs have previously been shown to induce anti-trout laruci acid hydroxylase immunoreactive proteins in bluegill and catfish. In this investigation, induction of lauric acid hydroxylase activity and immunoreactive proteins was confirmed, and the mass spectral analysis of specific hydroxylation products was performed in order to identify possible species-specific differences in fatty acid metabolism. Male bluegill (Lepomis macrochirus) and channel catfish (Ictalurus punctatus) were administered clofibrate or ciprofibrate 48 hr prior to hepatic or trunk kidney (catfish only) microsome preparation. While no significant differences were observed in male catfish, male bluegill had significant decreases in hematocrit and plasma protein indicating hemodilution due to ...

1995-12-31

222

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

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

2008-09-15

223

Spark decomposition of SF/sub 6/

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

1986-03-01

224

Oil-shale Fischer assay analysis from selected coreholes in the Piceance Creek Basin, Colorado. Data file

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 by-products 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 SF/sub 6/, spark-decomposed under specific experimental conditions, can produce cell death. Chemical analysis of spark-decomposed SF/sub 6/ has identified the major decomposition pathways and by-products. Biological testing of individual by-products or by-product mixtures has indicated that these major decomposition products may not account for the majority of the cell killing effects seen in our assays. Further experiments have suggested that S/sub 2/F/sub 10/ may be produced and accumulate under our specific ...

1989-07-01

225

Observations on the contributions of environmental restraints and innate stem cell ability to hematopoietic regeneration

A magnetic tape consisting of oil-shale Fischer assay data for selected well cores in the Piceance Creek Basin was prepared as a supplement to NTIS tape PB-230 607/AS. Both tapes serve as a data base for calculating oil-shale resources (Pitman, 1981). The tape is in standard IBM format and has the following specifications: EBCDIC (IBM standard), card-image, blocksize 1720 (blocked 20 to 1), record length 86, odd parity, 1,600 bpi, 9-track, no label tape. Each data file on the tape consists of records containing oil-shale analyses for one well core. The first record in a data file consists of a well-core identifier which is a three-digit number preceded by the character 'C'. The second record contains the well name and location, and the remaining records contain Fischer assay data.

1981-01-01

226

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

A competitive repopulation assay utilizing chromosome markers was used to assay the reconstituting potential of hematopoietic populations. The test populations consisted of tibial murine marrow locally irradiated with doses ranging from 1.5 Gy to 8.5 Gy and of marrow generated from either murine splenic or marrow stem cells. The purpose of this assay was to assess the innate proliferative potential and microenvironmental influences on the ability to repopulate. Regardless of origin, spleen repopulating ability consistently agreed with spleen colony-forming unit (CFU-s) content. Doses of radiation from 5 Gy to 8.5 Gy diminished, by a factor of 2, the ability to repopulate marrow despite maintenance of CFU-s levels. Marrow generated from splenic stem cells had one-fifth the repopulating ability of marrow derived from marrow stem cells, even though CFU-s levels were equivalent. The results imply that the splenic environment ...

1988-03-01

227

Development of a methodology to assess organometallic effects on bioenergetic systems

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

2011-05-31

228

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

A methodology for assessing the impact of subacute concentrations of organometallic agents on bioenergetic and oxidative damage processes in animals, cells and energy transducing subcellular organelles is being developed. Several of the assays are noninvasive and thus lend themselves to human tests. At the whole-animal level we utilize a treadmill chamber where physiological parameters of exercising animals are monitored. These include parameters of whole animals' work performance such as oxygen consumption, carbon dioxide evolution and endurance. Oxidative damage can be monitored in experiments by analyzing expired air of the animals for ethane and n-pentane. These alkanes correlate with lipid peroxidation in vivo. At the cellular and subcellular levels, respiratory activity, lipid peroxidation and free radical species are assayed. Respiratory activity is measured in muscle homogenates and isolated mitochondria using substrates which ...

1981-06-01

229

Use of flow cytometry to compare the antimicrobial efficacy of silver-containing wound dressings against planktonic Staphylococcus aureus and Pseudomonas aeruginosa

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

1995-12-01

230

British Library Electronic Table of Contents (United Kingdom)

Abstract Silver-impregnated wound dressings continue to be routinely used for the management of infected wounds, or wounds that are at risk of becoming infected. The ability of antimicrobials that have been incorporated into wound dressings to kill microorganisms within the dressing requires appropriate evaluation using in vitro models. In vitro models that have been exploited for this purpose have included the corrected zone of inhibition and the log reduction assay. However, these and other related culturable-based assays are purported to have poor correlation with the overall microbicidal barrier activity of an antimicrobial wound dressing. This is because culturable-based methods only retrospectively indicate bacterial cell death and do not take into account viable but nonculturable st...

2011-01-01

231

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

British Library Electronic Table of Contents (United Kingdom)

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

2006-01-01

232

SZ-685C, a marine anthraquinone, is a potent inducer of apoptosis with anticancer activity by suppression of the Akt/FOXO pathway

British Library Electronic Table of Contents (United Kingdom)

Background and purpose: The aims of this study were to investigate the anti-cancer activity of SZ-685C, an anthracycline analogue isolated from marine-derived mangrove endophytic fungi, and to explore the molecular mechanisms underlying such activity. Experimental approach: The effect of SZ-685C on the viability of cancer cell lines was investigated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. SZ-685C-induced apoptosis was assessed by Annexin V-fluorescein isothiocyanate/propidium iodide staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling assay and analysis of caspase activation. The effect of SZ-685C on the Akt/FOXO pathway was studied using Western blotting analysis, and the in vivo anti-tumour efficacy was examined in ...

2010-01-01

233

Probiotic-Induced Priming of Innate Immunity to Protect Against Rotaviral Infection

British Library Electronic Table of Contents (United Kingdom)

Understanding of probiotic-induced regulatory gene expression and networking is critical to further explore their roles in controlling infection. Transcriptional profile of selected innate immune genes in primary bovine intestinal epithelial cells was assessed over a time course of incubation with the probiotic Lactobacillus plantarum 299v. Based on gene expression results, a time point was chosen to prime epithelial cells with the probiotic prior to infection with rotavirus. Plaque assays and genomic analysis provided the basis for establishing the efficacy of probiotics in preventing a rotaviral infection. Plaque assays revealed that the probiotic is capable of decreasing (at least by 100-fold) the levels of live virus when the cells were primed with the probiotic. Results from gene expr...

2010-01-01

234

Optimized assay and storage conditions for enzyme activity profiling of ectomycorrhizae

British Library Electronic Table of Contents (United Kingdom)

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

2011-01-01

235

Nickel induced structural and functional alterations in mouse Leydig cells in vitro

British Library Electronic Table of Contents (United Kingdom)

The present study was aimed at investigating effects of nickel (NiCl2) on secretion of testosterone (T), cell viability, ultrastructure and apoptosis in mouse Leydig cells. Testosterone release was measured after 48h of culture with 15.67, 31.25, 62.5, 125, 250, 500 and 1000mmol/L NiCl2 or without NiCl2 using radioimmunoassay. Cell viability was assessed by a MTT (metabolic activity assay). Quantification of apoptotic cells was performed using TUNEL assay and the ultrastructural changes were analyzed using transmission electron microscopy. The viability was decreased after addition of >=250mmol/L NiCl2. A concentration-dependent depression of T production was observed. The percentage of apoptotic cells was significantly increased only after addition of 125, 250 and 1000mmol/L NiCl2. After ...

2011-01-01

236

Microtiter plate assay for phosphate using a europium-tetracycline complex as a sensitive luminescent probe

Limitation of immunohistochemical localization of estrogen receptor

A new luminescent europium probe is presented for the determination of phosphate (P) in microtiter plate format. The assay is based on the quenching of the luminescence of the europium-tetracycline (EuTc) 1:1 complex by phosphate using a reagent concentration of 20.8 {mu}mol/L. The probe is excited at 400 nm and displays a large Stokes' shift of 210 nm. The emission maximum is located at 616 nm. The system works best at neutral pH 7 and is therefore suitable for phosphate determination in biological and biochemical systems. The linear range of the calibration plot is from 5 x 10{sup -6} mol/L to 7.5 x 10{sup -4} mol/L of phosphate, and the limit of detection is 3 {mu}mol/L.

2006-01-12

237

Follow-up of pregnant women exposed to chicken pox: an audit of relationship between level of antibody and development of chicken pox

Although the radioreceptor method is widely used for estrogen receptor assay in human tissue, it has several limitations and a number of alternative methods are being explored. An immunohistochemical method of estrogen receptor assay using a specific antibody to estradiol has been proposed as a suitable alternative. The present study was designed to evaluate the validity of this method in detecting true estrogen receptors in human tumor tissue. Using radioiodinated antibody to estrogen, we have demonstrated that the estrogen antibody can detect the estrogen when it is bound to 4S type receptor but is unable to bind to estrogen when the hormone is bound to 8S type receptor. Our observations suggest that the immunohistochemical method of detection of intracellular cytosolic receptor for estrogen is not a suitable alternative to the currently used radioreceptor method.

1986-10-01

238

British Library Electronic Table of Contents (United Kingdom)

The purpose of this study was to validate through natural exposure a cut-off level of varicella zoster IgG as protective against infection with varicella zoster virus (VZV). Laboratory testing to determine VZV immune status of pregnant women exposed to varicella is recommended. Quantitative assays are now available which are sensitive and specific. More than 200 consecutive requests for screening in pregnant patients with recent varicella contacts were followed-up by questionnaire. DiaSorin LIAISON and VZV time resolved fluorescence immuno assay (VZV TRFIA) were used to measure VZV antibody level. One hundred fifty out of 209 (72%) questionnaires were returned; 14 patients developed varicella, 129 did not and seven were not known. Patients who had been given VZIG and developed varicella on...

2011-01-01

239

Evaluation of a commercial MRSA assay when multiple MRSA strains are causing epidemics

British Library Electronic Table of Contents (United Kingdom)

Rapid and reliable diagnostic methods are needed to control methicillin-resistant Staphylococcus aureus (MRSA) transmission. We studied the BD GeneOhm? MRSA Assay which is based on one specific amplification product at the junction of the right extremity sequence of the staphylococcal cassette chromosome mec (SCCmec) and the chromosomal sequence of orfX of S. aureus. The test was applied on 95 clinical isolates in Finland: 83% were positive. The isolates giving negative results represented several pulsed-field gel electrophoresis (PFGE) types and harboured SCCmec types IV, V, VI or were new types with different combinations of ccr genes.

2009-01-01

240

Estimates of quantitative genetic parameters of immunological traits in the chicken.

Establishment of a method for RIA of transferrin

Three in vivo assays were used to measure the immunocompetence of chickens in two generations of a selection experiment. The obtained data were used to estimate the variance components for sire and dam for antibody production to Pasteurella multocida and Mycoplasma gallisepticum vaccines, for T-cell-mediated immunity evaluated by a phytohemagglutinin wing web assay, and for clearance of foreign particles from the circulatory system. Heritabilities of and genetic correlations among these immunological traits were calculated from the sire variance components. Heritability estimates of the immunological traits based on the sire component of variance ranged from .06 to .53, and genetic correlations among immunological traits were generally negative. PMID:1956847

1991-10-01

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241

Detection of Visual Deficits in Aging DBA/2J Mice by Two Behavioral Assays

Objective: To develop a method for RIA of transferrin without dilution of the serum sample. Methods: The measurable range was broadened through: Lowering of the specific radioactivity of the labelled antigen, increasing the concentration of the anti-serum and improve the curve fitting method. Results: The antiserum affinity was 3.82 x 10"-"1"0 mol/L, the specific radioactivity of the labelled antigen was 15.3 mg/mic, inter-assay error CV 5.38%, intra-assay error CV = 3.5% and the shape of the curve was good. For 30 normal control specimens, the mean TF content was 2.81 +- 0.972 mg/ml. Conclusion: Development of a method for RIA of TF without previous dilution of the test serum was of clinical value

2002-04-01

242

British Library Electronic Table of Contents (United Kingdom)

Purpose: The DBA/2J mice have been used as an animal model for human pigmentary glaucoma. However, these mice develop various degrees of disease symptoms at different ages, making it difficult to detect pathological changes of retinal degeneration at glaucoma onset. The purpose of this study is to develop a non-invasive assay to identify individual mice that develop visual deficits. Materials and Methods: We apply two behavioral tests, a swimming test of visual discrimination and a test of optomotor response, to identify glaucomatous DBA/2J mice. We then examine whether the elevation of intraocular pressure (IOP), the common risk factor for glaucoma, affects visual performances of the DBA/2J mice. We further compare the retinal ganglion cell death, one of the signature glaucoma symptoms, i...

2011-01-01

243

Clinical and experimental studies of octocrylene's allergenic potency

British Library Electronic Table of Contents (United Kingdom)

Background. Reports of positive patch test and photopatch test reactions to the chemical ultraviolet filter octocrylene have increased during the last decade. Little is known about the reason for octocrylene's allergenic activity. Objectives. To present and discuss the results of patch tests and photopatch tests with octocrylene, and to investigate the possible cause of its allergenic properties. Methods. Results of patch tests and photopatch tests with octocrylene in patients with adverse skin reactions to sunscreen products and/or ketoprofen were collected. The allergenic potency of octocrylene was investigated in the murine local lymph node assay (LLNA). Chemical reactivity assays were used to mimic octocrylene's interaction with biomolecules. Results. We report 23 cases of positive tes...

2011-01-01

244

Cell-mediated mutagenesis and cell transformation of mammalian cells by chemical carcinogens. [Rats, hamsters

Antibacterial iridoid glucosides from Eremostachys laciniata

We have developed a cell-mediated mutagenesis assay in which cells with the appropriate markers for mutagenesis are co-cultivated with either lethally irradiated rodent embryonic cells that can metabolize carcinogenic hydrocarbons or with primary rat liver cells that can metabolize chemicals carcinogenic to the liver. During co-cultivation, the reactive metabolites of the procarcinogen appear to be transmitted to the mutable cells and induce mutations in them. Assays of this type make it possible to demonstrate a relationship between carcinogenic potency of the chemicals and their ability to induce mutations in mammalian cells. In addition, by simultaneously comparing the frequencies of transformation and mutation induced in normal diploid hamster cells by benzo(a)pyrene (BP) and one of its metabolites, it is possible to estimate the genetic target size for cell transformation in vitro.

1977-01-01

245

British Library Electronic Table of Contents (United Kingdom)

Eremostachys laciniata (L) Bunge (family: Lamiaceae alt. Labiatae; subfamily: Lamioideae) is one of the 15 endemic Iranian herbs of the genus Eremostachys. A decoction of the roots and flowers of E. laciniata has traditionally been taken orally for the treatment of allergies, headache and liver diseases. Three antibacterial iridoid glucosides, phloyoside I (1), phlomiol (2) and pulchelloside I (3) have been isolated from the rhizomes of this plant. The structures of these compounds were elucidated unequivocally by a series of 1D and 2D NMR analyses. The antibacterial activity and brine shrimp toxicity of these compounds were assessed using the resazurin microtitre assay and the brine shrimp lethality assay, respectively. All three iridoid glycosides 1-3 exhibited from low to moderate level...

2009-01-01

246

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

[Hormonal disorders in chronic alcoholic intoxication].

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

247

The Significance of Clopidogrel Low-Responsiveness on Stent Thrombosis and Cardiac Death Assessed by the Verifynow P₂Y₁₂ Assay in Patients With Acute Coronary Syndrome Within 6 Months After Drug-Eluting Stent Implantation

Hormone levels were examined in the venous blood in 54 men suffering from stage 2 alcoholism and in 30 normal subjects, using a radioimmunochemical assay. The alcoholics were found to have a statistically significant increase in prolactin and a decrease in testosterone. A definite ratio in the secretion of these hormones differing from that in the control group was elucidated. This ratio (prolactin-testosterone index) is proposed as a diagnostic test of alcoholism. PMID:6441404

1984-01-01

248

Synthesis of b-functionalized Temoporfin derivatives for an application in photodynamic therapy

Background and ObjectivesClopidogrel resistance or low-responsiveness may be associated with recurrent atherothrombotic events after drug-eluting stent (DES) implantation. We prospectively...Full Text Available

2009-12-01

249

British Library Electronic Table of Contents (United Kingdom)

The synthesis of novel b-functionalized derivatives of the clinically used photosensitizer Temoporfin has been achieved by nucleophilic addition reactions to a corresponding diketo chlorin. The b-substituted dihydroxychlorin products exhibit a strong absorption in the red spectral region, a high singlet oxygen quantum yield, and were found to be highly effective in in vitro assays against HT-29 tumor cells.

2011-01-01

250

Side chain SAR of bicyclic [beta]-lactamase inhibitors (BLIs). 1. Discovery of a class C BLI for combination with imipinem

Serum Globotriaosylceramide Assay as a Screening Test for Fabry Disease in Patients with ESRD on Maintenance Dialysis in Korea

Bridged monobactam {beta}-lactamase inhibitors were prepared and evaluated as potential partners for combination with imipenem to overcome class C {beta}-lactamase mediated resistance. The (S)-azepine analog 2 was found to be effective in both in vitro and in vivo assays and was selected for preclinical development.

2010-09-17

251

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

Background/AimsFabry disease is an X-linked recessive and progressive disease caused by α-galactosidase A (α-GaL A) deficiency. We sought to assess the prevalence...Full Text Available

2010-12-01

252

No difference in antibody titers against xenotropic MLV related virus in prostate cancer cases and cancer-free controls

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

1984-04-15

253

British Library Electronic Table of Contents (United Kingdom)

New ELISA assays were developed to measure immunoreactivity for XMRV. Antibody titers were measured in a cohort of prostate cancer cases and cancer free controls from the central United States. No statistically significant differences were observed in immunoreactivity between cases and controls for either the XMRV-env or the XMRV-gag antigen.

2011-01-01

254

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

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

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

1987-06-01

255

Hair root versus red cell individual phenotype in Sardinian heterozygotes for G6PD deficiency (Mediterranean type).

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

1986-06-01

256

Gamma scanning of FBTR fuel pins

G6PD activity was assayed in 20 Sardinian heterozygotes for G6PD deficiency and related to that of LDH and MDH. One of these heterozygotes showed a deficient phenotype in all her follicles, while the...Full Text Available

1976-09-01

257

Experience with Aerosol Generation During Rotary Mode Core Sampling in the Hanford Single Shell Waste Tanks

This paper presents the results obtained in the gamma scanning of two fuel pins from the bent subassembly of the fast breeder test reactor (FBTR) using a segmented gamma scanning system employing segment correlation developed for the assay of glove box solid waste. In addition to the actinide profiles, the paper also discusses the fission products and clad activation product profiles and tries to correlate the experimental values of the latter with computed values. (author). 4 refs., 1 fig., 1 tab.

258

Evaluation of Sanofi Diagnostics Pasteur Chlamydia Microplate EIA shortened assay and comparison with cell culture and Syva Chlamydia MicroTrak II EIA in high- and low-risk populations.

This document provides data on aerosol concentrations in tank headspaces, total mass of aerosols in the tank headspace, and mass of aerosols sent to the exhauster during rotary mode core sampling from November 1994 through June 1999. A decontamination factor for the RMCS exhauster filter housing is calculated based upon operational data and non-destructive assay.

2001-03-23

259

Discrimination of Listeria monocytogenes from other Listeria species by ligase chain reaction.

Seven hundred thirty-two female urogenital samples were collected for Chlamydia trachomatis testing by both the Sanofi Diagnostics Pasteur (Chaska, Minn.) Chlamydia Microplate EIA by the shortened protocol...Full Text Available

1995-11-01

260

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

A ligase chain reaction assay based on a single-base-pair difference in the V9 region of the 16S rRNA gene (16S rDNA) was developed to distinguish between Listeria monocytogenes and other Listeria species....Full Text Available

1992-11-01

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261

Detection of pathogenic Yersinia enterocolitica in foods and water by immunomagnetic separation, nested polymerase chain reactions, and colorimetric detection of amplified DNA.

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

1994-05-01

262

Detection of drugs and explosives using neutron computerized tomography and artificial intelligence techniques

A two-step polymerase chain reaction (PCR) procedure with two nested pairs of primers specific for the yadA gene of Yersinia enterocolitica was developed. The PCR assay identified all common pathogenic...Full Text Available

1993-09-01

263

Dengue Plaque Reduction Neutralization Test (PRNT) in Primary and Secondary Dengue Virus Infections: How Alterations in Assay Conditions Impact Performance

In this study the development of a methodology to detect illicit drugs and plastic explosives is described with the objective of being applied in the realm of public security. For this end, non-destructive assay with neutrons was used and the technique applied was the real time neutron radiography together with computerized tomography. The system is endowed with automatic responses based upon the application of an artificial intelligence technique. In previous tests using real samples, the system proved capable of identifying 97% of the inspected materials.

2010-06-01

264

Comparison of C₁₈-Carboxypropylbetaine and Glass Bead DNA Extraction Methods for Detection of Mycobacterium bovis in Bovine Milk Samples and Analysis of Samples by PCR

Dengue virus (DENV) infection is a worsening global health problem. The plaque reduction neutralization test (PRNT) is currently considered to be the “gold standard” to characterize...Full Text Available

2009-11-01

265

Comparative Study of ¹²⁵I- and [³H]Acetate-Labeled Antibodies in Detecting Iridescent Viruses

The purpose of this prospective study was to compare two different milk preparation methods to assay for the presence of Mycobacterium bovis by PCR. Detection by a C₁₈-carboxypropylbetaine...Full Text Available

1998-08-01

266

Commercially Available Antibodies to Human Tumour Necrosis Factor-? Tested for Cross-Reactivity with Ovine and Bovine Tumour Necrosis Factor-? using Flow Cytometric Assays

Radioimmunoassays for detecting cell-associated or released virus are described using either ¹²⁵I- or [³H]acetate-labeled antibodies. In the first assay system, antigen-antibody...Full Text Available

1974-07-01

267

An immunochemical approach to the study of DNA damage and repair

A thorough understanding of the immune system, including the role of different cytokines, during inflammatory diseases in ruminants could lead to the development of new diagnostic methods and treatments....Full Text Available

2004-01-01

268

Amp-PCR: Combining a Random Unbiased Phi29-Amplification with a Specific Real-Time PCR, Performed in One Tube to Increase PCR Sensitivity

The overall objective of this project is to produce antibodies to unique modified DNA bases and develop immunochemical assays to quantitate these lesions in damaged DNA. During this past year we have developed an antibody and chemical test to quantitate a basic sites in DNA and produced antibodies to the 8-oxopurines. This report discusses the detection of a basic sites in DNA and the preparation of antibodies to 8-hydroxyadenine and 8-hydroxyguanine.

1990-11-14

269

Absolute fission rates in the FFTF

In clinical situations where a diagnostic real-time PCR assay is not sensitive enough, leading to low or falsely negative results, or where detection earlier in a disease progression would benefit the...Full Text Available

270

Effects of alpha-deuterium substitution on the mutagenicity of 4-(methyl-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK)

The part of the FFTF Reactor Characterization Program reported in this paper is a measurement of absolute fission rates of eight major fuel isotopes at two different positions within the reactor. The instruments employed in these tests were fission ionization chambers for which the absolute efficiency and fissionable deposit mass assay have been rigorously established.

1981-06-01

271

Depuration of shellfish by irradiation: Final technical report, October 1, 1987--March 31, 1989

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a carcinogenic tobacco specific nitrosamine, can be converted to electrophilic diazohydroxide intermediates by metabolic hydroxylation of either the methylene carbon (carbon 4) or the methyl carbon attached to the nitrosamine group. To investigate the relative importance of these two processes in NNK mutagenesis, we synthesized 4,4-dideutero-4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone((4,4,-D2)NNK) and 4-(trideuteromethylnitrosamino)-1-(3-pyridyl)-1-butanone ((CD3) NNK), and evaluated their mutagenic activities in Salmonella typhimurium tester strains. In the presence of Aroclor induced rat liver 9000 g supernatant, NNK and (4,4-D2)NNK had comparable mutagenic activities towards S. typhimurium TA 1535 and TA 100, but (CD3)NNK was inactive in both strains. These results suggest that hydroxylation of the methyl group of NNK is more important than hydroxylation of carbon 4 in its activation to a mutagen. To test ...

1983-01-01

272

Chemical, sensory and microbiological changes of gamma irradiated coconut cream powder

Studies by the University of Lowell Radiation Laboratory and the US National Marine Fisheries Service N.E. Laboratory in Gloucester, MA on softshelled clams (Mya arenaria) demonstrated the effectiveness of low to medium doses of Cobalt 60 source gamma irradiation in the inactivation of Escherichia coli, Salmonella typhimurium, Staphylococcus aureus and Streptococcus faecalis. Post-irradiation survival and organoleptic studies when extended to hardshelled clams (Mercinaria mercenaria) and American oysters (Crassostrea virginica) showed no significant decline in consumer qualities or 6 day post irradiation survival in oysters at doses of up to 3.0 kGy. The capacities of the American oyster to sustain relatively high doses of gamma irradiation were demonstrated by 6 day post-exposure survivorship values of greater than 90% for samples receiving 3.0, 5.0 and 7.0 kGy. Initial studies of inactivation of Polio I virus and a simian rotavirus (SA-11) was conducted in both ...

273

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

A study was carried out to determine optimum decontamination dose for a locally manufactured coconut cream powder. Samples were gamma irradiated (0-15 kGy) and ageing process was achieved using GEER oven at 60 deg. C for 7 days, which is equivalent to one-year storage at room temperature. Iodine value (IV), ranging from 4.8 to 6.4, was not affected by radiation doses and storage, however peroxide value and thiobarbituric acid (TBA) generally increased with radiation doses. In most samples, peroxide value (meq/kg) reduced after storage, whilst the TBA (mg malonaldehyde/kg), indicator for product quality, slightly increased. The sensory evaluation conducted using 25 taste panellists indicated that scores on odour, creamy taste and overall acceptance for all irradiated samples at more than 5 kGy were significantly lower (P<0.05) than the control. However, the panellists could not detect any significant differences among the irradiation doses (P>0.05). All stored products ...

2007-11-15

274

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

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

1978-06-01

275

Quantification of antibiotic drug potency by a two-compartment radioassay of bacterial growth

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

276

Online immunoaffinity LC/MS/MS. A general method to increase sensitivity and specificity: How do you do it and what do you need?

The two-compartment radioassay for microbial kinetics based on continuous measurement of the {sup 14}CO{sub 2} released by bacterial metabolism of 14C-labeled substrate offers a valuable approach to testing the potency of antimicrobial drugs. By using a previously validated radioassay with gram-positive and gram-negative bacteria, a group of protein synthesis inhibitors was evaluated for their effect on microbial growth kinetics. All tested drugs induced changes in both the slopes and intercepts of the growth curves. An exponential growth model was applied to quantify the drug effect on the processes of bacterial {sup 14}CO{sub 2} liberation and cell generation. The response was measured in terms of a generation rate constant. A linear dependence of the generation rate constant on the dose of spectinomycin was observed with Escherichia coli. Sigmoidal-shaped curves were found in the assays of chloramphenicol and tetracycline. The implications of dose-response ...

1990-06-01

277

Lyophilized human whole blood for internal and external quality assurance of lead in blood assays

There is an increased emphasis on hyphenated techniques such as immunoaffinity LC/MS/MS (IA-LC/MS/MS) or IA-LC/MRM. These techniques offer competitive advantages with respect to sensitivity and selectivity over traditional LC/MS and are complementary to ligand binding assays (LBA) or ELISA's. However, these techniques are not entirely straightforward and there are several tips and tricks to routine sample analysis. We describe here our methods and procedures for how to perform online IA-LC/MS/MS including a detailed protocol for the preparation of antibody (Ab) enrichment columns. We have included sample trapping and Ab methods. Furthermore, we highlight tips, tricks, minimal and optimal approaches. This technology has been shown to be viable for several applications, species and fluids from small molecules to proteins and biomarkers to PK assays. PMID:21872661

2011-08-22

278

Investigation of xenotropic murine leukemia virus-related virus (XMRV) in human and other cell lines.

Lyophilized human whole blood control material containing lead was prepared for internal and external quality assurance to evaluate and improve the analytical performance of lead. The samples were prepared in four different concentrations from outdated human whole blood stabilized with glucose and a citrate/phosphate buffer and provided under clean room conditions to avoid contamination. The lyophilized samples are easy to reconstitute with water. The materials were evaluated according to a statistical model. The lead concentrations in the specimens are close to blood lead levels usually following environmental and industrial exposure, particular in occupational health to the control of lead exposure at work regulations. The materials are available to the commercial as well as the scientific community. The Danish External Quality Assessment Scheme (DEQAS) for lead in blood is intended to complement the internal quality control for the Danish laboratories assaying ...

1990-10-01

279

Feasibility of subcriticality and NDA measurements for spent fuel by frequency analysis techniques with {sup 252}Cf

Xenotropic murine leukemia virus-related virus (XMRV) was discovered in human prostate tumors and later in some chronic fatigue syndrome (CFS) patients. However, subsequent studies have identified various sources of potential contamination with XMRV and other murine leukemia virus (MLV)-related sequences in test samples. Biological and nucleotide sequence analysis indicates that XMRV is distinct from known xenotropic MLVs and has a broad host range and cell tropism including human cells. Therefore, it is prudent to minimize the risk of human exposure to infection by evaluating XMRV contamination in cell lines handled in laboratory research and particularly those used in the manufacture of biological products. Nested DNA PCR assays were optimized for investigating XMRV gag and env sequences in various cell lines, which included MRC-5, Vero, HEK-293, MDCK, HeLa, and A549, that may be used in the development of some vaccines and other cell lines broadly used in ...

2011-10-11

280

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

The {sup 252}Cf-source-driven frequency analysis method can be used for measuring the subcritical neutron multiplication factor of arrays of LWR fuel and as little as a single PWR fuel assembly. These measurements can be used to verify the criticality safety margins of spent LWR fuel configurations and thus could be a means of obtaining the information to justify burnup credit for spent LWR transportation/storage casks. In addition, the data can be used to validate calculational methods for criticality safety. These measurements provide parameters that have a higher sensitivity to changes in fissile mass than neutron multiplication factor and thus serve as a better test of calculational methods. The analysis have also shown that measurement of the cross power spectral density (CPSD) between detectors on one side of a single fuel assembly and an internal or external {sup 252}Cf source driving the fission chain multiplication process can be used for nondestructive ...

1996-05-01

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281

Differentiation to adipocytes in accompanied by an increase in the amounts of Gi- and Go-proteins in 3T3-L1 cells

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

282

Consistent effects of a major QTL for thermal resistance in field-released Drosophila melanogaster.

Treatment of cultures of 3T3-L1 cells with methylisobutyl-xanthine and dexamethasone has been shown to result in accumulation of lipid and conversion to the morphology of adipocytes in more than 90% of the cells. The status of the stimulatory (Gs), inhibitory (Gi) and Go-proteins during the course of 3T3-L1 differentiation was examined. The amount of alpha subunit of Gs (#alpha#Gs), assayed by radiolabeling in the presence of cholera toxin and ["3"2P]NAD"+, increased upon differentiation as previously described by others. The amounts of #alpha#Gi and #alpha#Go assayed by radiolabeling in the presence of pertussis toxin and ["3"2P]NAD"+ increased 3-fold upon differentiation. Immunoblots of cell membranes subjected to gel electrophoresis in sodium dodecyl sulfate were probed with two rabbit antisera raised against bovine brain #alpha#Go and with one raised against the#beta#-subunit of the bovine rod-outer-segment G-protein, referred to as ...

1986-06-08

283

Age sensitivity of juvenile mussels (Utterbackia imbeciles Say) to copper and cadmium exposure

Molecular genetic markers can be used to identify quantitative trait loci (QTL) for thermal resistance and this has allowed characterization of a major QTL for knockdown resistance to high temperature in Drosophila melanogaster. The QTL showed trade-off associations with cold resistance under laboratory conditions. However, assays of thermal tolerance conducted in the laboratory may not necessarily reflect performance at varying temperatures in the field. Here we tested if lines with different genotypes in this QTL show different thermal performance under high and low temperatures in the field using a release recapture assay. We found that lines carrying the QTL genotype for high thermal tolerance were significantly better at locating resources in the field releases under hot temperatures while the QTL line carrying the contrasting genotype were superior at cold temperatures. Further, we studied copulatory success between the different QTL ...

2011-06-15

284

A DC-81-indole conjugate agent suppresses melanoma A375 cell migration partially via interrupting VEGF production and stromal cell-derived factor-1a-mediated signaling

In recent years, there has been increased interest in using early life stages of freshwater bivalves to test the toxicity of dissolved constituents in water. The authors have developed laboratory and in situ assays with artificially cultured Utterbackia imbecillis to examine effects of contaminants and existing conditions in embayments and rivers on mortality and reproductive success of unionid mussels. Age sensitivity of U. imbecillis to Cd and Cu was examined using both static acute and 8-day static renewal bioassays. Both aqueous and sediment exposures demonstrated greater sensitivity to Cd than Cu. LC{sub 50}`s for two-day old (2d) and 9d mussels were approximately half the LC{sub 50} calculated for 16d mussels in 48-hr bioassays with Cd. Additionally, acute assays were repeated twice to examine variability in response between different mussel cultures to the two metals. The results of this work reinforce the potential of juvenile U. ...

1994-12-31

285

British Library Electronic Table of Contents (United Kingdom)

Pyrrolo[2,1-c][1,4]benzodiazepine (PBD) chemicals are antitumor antibiotics inhibiting nucleic acid synthesis. An indole carboxylate-PBD hybrid with six-carbon spacer structure (IN6CPBD) has been previously demonstrated to induce melanoma cell apoptosis and reduce metastasis in mouse lungs. This study aimed at investigating the efficacy of the other hybrid compound with four-carbon spacer (IN4CPBD) and elucidating its anti-metastatic mechanism. Human melanoma A375 cells with IN4CPBD treatment underwent cytotoxicity and apoptosis-associated assays. Transwell migration assay, Western blotting, and ELISA were used for mechanistic study. IN4CPBD exhibited potent melanoma cytotoxicity through interrupting G1/S cell cycle progression, increasing DNA fragmentation and hypodipoidic DNA contents, a...

2011-01-01

286

The effects of gamma irradiation of food contact plastics on the extent of migration of hindered phenol antioxidants into fatty food simulants

Solid-phase synthesis and pharmacological evaluation of novel nucleoside-tethered dinuclear platinum(II) complexes

In view of the interest in the use of gamma irradiation to extend the storage life of pre-packaged foods a report is made on the effects of gamma radiation on the migration of Irganox 1076 and Irganox 1010, antioxidants present in food packaging plastics, into synthetic triglyceride fatty-food simulant HB307 and iso-octane. The extent of migration was assayed by h.p.l.c. techniques. Migration into HB307 and iso-octane decreased steadily as irradiation progressed suggesting that gamma irradiation of packaging materials helps to prevent the contamination of food by antioxidants in the packaging. (U.K.).

287

British Library Electronic Table of Contents (United Kingdom)

Three novel inosine-based dinuclear platinum complexes have been synthesized via a solid-phase strategy. In these compounds, the metal is linked both to the N-7 of the purine nucleus and to the terminal amine group of a hexylamine side chain installed on N-1. Cis- or trans- diamine as well as ethylenediamine ligands are coordinated to platinum along with a chloride. The synthesised complexes were tested against four different human tumor cell lines. One of these complexes proved to be more cytotoxic than cisplatin against the MCF7 cancer cell line in a short-term exposure assay.

2011-01-01

288

Radiolabelling of. cap alpha. -bungarotoxin to high specific radioactivity with complete retention of biological activity

Quality engineering and control. Semiannual progress report, May-October 1979

Radioiodinated ..cap alpha..-bungarotoxin, the venom of Bungarus multicinctus, is commonly used for the detection and quantification of nicotinic acetylcholine receptors (nAChR) and anti-nAChR antibodies. A radioiodination method is described which is simple and yields a fully active product which, after fractionation, has a specific activity tenfold higher (900 Ci/mmol) than those routinely used, resulting in enhanced sensitivity of assays and conservation of material, especially muscle extracts.

1984-10-01

289

Mitochondrial DNA variants in Drosophila melanogaster are expressed at the level of the organismal phenotype.

The following are reported: colorimetric determination of Ti in Cr, decontamination of metals by anodic dissolution, computer code for x-ray peak identification, gel-permeation chromatography of CoRezyn 1664 polyester resin, infrared monitoring of gases and vapors, particle size analysis of dusts, nondestructive analysis of Oralloy for removable alpha contamination, dissolution (digestion) of high-fired oxides, assay of U in a Pu-U alloy, measurement of oxide thickness on Pu metal by x-ray diffraction, analysis of precipitated Pu peroxide, differentiation of Dowex SBR and 11 resins, and determination of Pu solubility in simulated lung fluid.

290

Effect on the reproductive functions of female rhesus monkeys of feeding irradiated wheat flour and potato diet

A plethora of experimental studies use mtDNA as a marker of demographic processes without questioning the possibility that selection may bias their interpretations. We studied four lines of Drosophila melanogaster that have a standardized nuclear DNA but variable mtDNA. We completed the sequencing of the mitochondrial genomes (excluding the A+T rich region) and compiled the differences. We then assayed male influence on oviposition, starvation resistance, lipid proportion and physical activity. We discuss these results in terms of the known differences between the lines and conclude that naturally occurring mtDNA variants in D. melanogaster are expressed at the level of the organismal phenotype. PMID:21757031

2011-07-05

291

Effect of synthetic detergents on germination of fern spores

The effects on reproductive functions of feeding adult rhesus monkeys on a diet of irradiated wheat flour and potato was investigated. Wheat flour and potatoes were exposed to 75 000 and 10 000 rad, respectively, of #gamma#-irradiation and then fed to the animals. Various parameters of reproductive function were compared with those of animals fed on unirradiated food. These were: external changes in the reproductive organs, menstrual cycle, vaginal cytology, assay of urinary hormones (oestrogens and pregnanediol), and post-mortem histopathological examination of the reproductive organs. At the end of 3 months, the experimental and control animals behaved similarly with regard to their reproductive functions. (author).

292

Chromatographic separation of lithium isotopes by hydrous manganese(IV) oxide

Synthetic detergents constitute one of the most important water pollutants by contaminating the lakes and rivers through domestic and industrial use. Considerable information is now available for the adverse effects of detergents an aquatic fauna including fish, algae, and higher aquatic plants. Marked inhibition of germination in orchids and brinjals and of seedlings growth in raddish suggest that rapidly growing systems could be sensitive to detergent polluted water. The present study of the effect of linear alkyl benzene sulphonate on germination of the spores of a fern, Diplazium esculentum aims at the understanding of the effects of water pollution on pteridophytes and the development of spore germination assay for phytoxicity evaluation.

1986-12-01

293

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

Separation of lithium isotopes was investigated by chemical ion exchange with a hydrous manganese(IV) oxide ion exchanger using an elution chromatography. The capacity of manganese(IV) oxide ion exchanger was 0.5 meq/g. The heavier lithium isotope was enriched in the solution phase, while the lighter isotope was enriched in the ion exchanger phase. The separation factor was determined according to the method of Glueckauf from the elution curve and isotopic assays. The separation factor of {sup 6}Li{sup +} -{sup 7}Li{sup +} isotope pair fractionation was 1.018.

2001-06-01

294

Biodistribution of phenylboric acid derivative entrapped lipiodol and 4-borono-2-{sup 18}F-fluoro-L-phenylalanine-fructose in GP7TB liver tumor bearing rats for BNCT

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

295

Applications of nanotechnology in food packaging and food safety: Barrier materials, antimicrobials and sensors

A new phenylboric acid derivative entrapped lipiodol (PBAD-lipiodol) was developed as a boron carrier for the boron neutron capture therapy (BNCT) of hepatoma in Taiwan. The biodistribution of both PBAD-lipiodol and BPA-fructose was assayed in GP7TB hepatoma-bearing rat model. The highest uptake of PBAD-lipiodol was found at 2 h post injection. The application of BNCT for the hepatoma treatment in tumor-bearing rats is suggested to be 2-4 h post PBAD-lipiodol injection.

2010-03-15

296

British Library Electronic Table of Contents (United Kingdom)

In this article, several applications of nanomaterials in food packaging and food safety are reviewed, including: polymer/clay nanocomposites as high barrier packaging materials, silver nanoparticles as potent antimicrobial agents, and nanosensors and nanomaterial-based assays for the detection of food-relevant analytes (gasses, small organic molecules and food-borne pathogens). In addition to covering the technical aspects of these topics, the current commercial status and understanding of health implications of these technologies are also discussed. These applications were chosen because they do not involve direct addition of nanoparticles to consumed foods, and thus are more likely to be marketed to the public in the short term.

2011-01-01

297

Antimicrobial potential of Glycyrrhiza glabra roots

British Library Electronic Table of Contents (United Kingdom)

The present study was aimed to investigate antimicrobial potential of Glycyrrhiza glabra roots. Antimycobacterial activity of Glycyrrhiza glabra was found at 500mg/mL concentration. Bioactivity guided phytochemical analysis identified glabridin as potentially active against both Mycobacterium tuberculosis H37Ra and H37Rv strains at 29.16mg/mL concentration. It exhibited antimicrobial activity against both Gram-positive and Gram-negative bacteria. Our results indicate potential use of licorice as antitubercular agent through systemic experiments and sophisticated anti-TB assay.

2008-01-01

298

Two subpopulations of stem cells for T cell lineage

Synthesis of 4,5,6,7 and 2,4,5,6,7 deuterium-labeled indole-3-acetic acid for use in mass spectrometric assays

An assay system for the stem cell that colonizes the thymus and differentiates into T cells was developed, and by using this assay system the existence of two subpopulations of stem cells for T cell lineage was clarified. Part-body-shielded and 900-R-irradiated C57BL/6 (H-2b, Thy-1.2) recipient mice, which do not require the transfer of pluripotent stem cells for their survival, were transferred with cells from B10 X Thy-1.1 (H-2b, Thy-1.1) donor mice. The reconstitution of the recipient's thymus lymphocytes was accomplished by stem cells in the donor cells and those spared in the shielded portion of the recipient that competitively colonize the thymus. Thus, the stem cell activity of donor cells can be evaluated by determining the proportion of donor-type (Thy-1.1+) cells in the recipient's thymus. Bone marrow cells were the most potent source of stem cells. By contrast, when the stem cell activity was compared between spleen ...

1985-11-01

299

Surgical adjuvant therapy in colon carcinoma: a human tumor spheroid model for evaluating radiation sensitizing agents

Syntheses are described for tetra and pentadeutero indole-3-acetic acid (IAA) labeled in positions 4,5,6,7 or 2,4,5,6,7 of the indole moiety. Polydeuterated IAA is proposed as an internal standard for gas chromatographic-mass spectrometric analysis of IAA by selected ion monitoring. Nanogram amounts of IAA may be assayed by monitoring the base peak of IAA at m/z = 130 (134 for d/sub 4/-IAA) and the molecular ion of the methyl ester of IAA at 189 (193 for d/sub 4/-IAA). Deuterium in positions 4,5,6, and 7 and, to only a slightly lesser extent, that in position 2 of IAA is retained during alkali treatment, thus permitting use of these compounds as internal standards for assay of IAA released by alkaline hydrolysis of ester and amide conjugates. The use of polydeutero internal standards separates the standards from the isotope cluster caused by the normal abundance of heavy isotopes and also permits use of reduced mass resolution, thus leading to ...

1980-10-01

300

Spark decomposition of SF/sub 6/: Chemical and biological studies

HT-29 human colon tumor cells growing as spheroids have been evaluated as a model system for measuring the response of human colon tumor cell to antineoplastic agents. HT-29 cells have been capacity to form spheroids up to 1 mm or more in diameter when grown in spinner culture. The multicellular HT-29 spheroids develop hypoxic centers reflecting the cellular conditions found in human cancer treatment, i.e., nutritionally deficient hypoxic cells that are felt to be a significant source of both radiation and chemotherapy clinical treatment failures. Spheroids of increasing size were radiated and then dispersed into single cells for colony survival assay. Compared with irradiated single cell suspensions, the spheroid cells demonstrated a significant increase in radioresistance. Growing spheroids developed a complex radiation survival curve which was variable with respect to size of the spheroid. The drug 5-Fu was studied to examine in a preliminary fashion its ...

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

301

Screening for von Willebrand disease: contribution of an automated assay for von Willebrand factor activity.

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 ...

1987-01-01

302

Screening for congenital hypothyroidism in Lebanon - An example of newborn screening in absence of mandatory law

Summary.? Measuring von Willebrand factor (VWF) activity is essential to the diagnosis of von Willebrand disease (VWD). The VWF activity is usually assessed based on measurement of the ristocetin cofactor (VWF:RCo). However, that test is technically challenging and has high intra- and inter-assay variabilities. The HemosIL VWF activity (VWF:AC) is a fully automated assay, recently proposed as a good alternative to VWF:RCo for VWD diagnosis. This study was undertaken to assess this new method. First, the analytical performance of VWF:AC on an automated coagulo-meter (ACLTop) was determined, and then this new method was compared with VWF:RCo and the platelet function analyzer (PFA100) for 160 patients referred for VWD screening. The VWF:AC achieved acceptable precision with within-run and between-run coefficients of variation ranging from 2.3% to 14.1%, and linearity from 10% to 100%. Despite some marked differences between VWF:AC and VWF:RCo for ...

2011-09-28

303

Radwaste Drum Assay Technology by Segmented Gamma Scanning System

This paper presents a group's efforts to introduce national newborn screening in Lebanon. Tandem mass spectrometry (MS) technology offers the prospect of utilizing filter paper blood specimens to screen newborns for several metabolic disorders. In order to be cost-effective, it is necessary to have a mandatory newborn screening policy. At present such a policy is not available in Lebanon. A national screening survey for congenital hypothyroidism (CH) was carried out to assess the incidence of this anomaly in the Lebanese population. Such an initiative is essential as a first step for assessing the feasibility and cost of a national screening programme. Coated beads, blood spot standards for quality control and buffer solutions used for assays were prepared in-house to minimize costs. At first, samples of umbilical cord blood spots were assayed for TSH using IRMA technique. Those samples showing borderline or high values of TSH were then ...

2004-10-01

304

Potent, selective and cell penetrant inhibitors of SF-1 by functional ultra-high-throughput screening.

Nuclear Act of Korea requires the manifest of low and intermediate level radioactive waste generated from nuclear power plants and other nuclear facilities prior to deliver to a disposal sites. Individual history records of the radioactive waste should be described the information about the activity of nuclides in the drum, total activity, weight, the type of waste. So a fully automated nuclide analysis assay system(non-destructive) of a radwaste drum was developed. For the nuclides that could not be analysed directly by MCA, the activities of the representative {gamma}-emitters(Cs-137, Co-60) contained in the drum were measured by using that system. Then the scaling factors were used to calculate the activities of {alpha} - and {beta}-emitters. This assay system divided a drum into 8 segments, a segment into 8 sectors to minimize analysis error, and used several methods(transmission ratio, differential peak absorption, mean density ...

2009-01-15

305

Double-antibody solid-phase radioimmunoassay: a simplified phase-separation procedure applied to various ligands

The steroidogenic factor 1 (SF-1, also known as NR5A1) is a transcription factor belonging to the nuclear receptor superfamily. Whereas most of the members of this family have been extensively characterized, the therapeutic potential and pharmacology of SF-1 still remains elusive. Described here is the identification and characterization of selective inhibitory chemical probes of SF-1 by a rational ultra-high-throughput screening (uHTS) strategy. A set of 64,908 compounds from the National Institute of Health's Molecular Libraries Small Molecule Repository was screened in a transactivation cell-based assay employing a chimeric SF-1 construct. Two analogous isoquinolinones, ethyl 2-[2-[2-(2,3-dihydro-1,4-benzodioxin-7-ylamino)-2-oxoethyl]-1-oxoisoquinolin-5-yl]oxypropanoate (SID7969543) and ethyl 2-[2-[2-(1,3-benzodioxol-5-ylmethylamino)-2-oxoethyl]-1-oxoisoquinolin-5-yl]oxypropanoate and (SID7970631), were identified as potent submicromolar inhibitors, yielding ...

2008-03-11

306

Double-antibody solid-phase radioimmunoassay: a simplified phase-separation procedure applied to various ligands

The purpose was to develop a simplified and reliable method of separating free from antibody-bound ligand using a precipitating antibody linked to a cellulose derivative. Dose-response curves and control sera were set up in parallel for various pituitary and placental polypeptides, steroid hormones, insulin, glucagon, triiodothyronine, thyroxine, angiotensin I, calcitonin, gastrin, cyclic AMP, and digoxin. After first-antibody reactions had reached equilibrium, free and bound ligand were separated using a double-antibody solid-phase system in parallel with conventional methods, including dextran-coated charcoal, double-antibody precipitation, single-antibody solid phase, organic solvents, salt precipitation, and anion-exchange resins. The effect of variations in temperature, incubation time, protein content, pH, and amount of separating material added were studied. The results showed that separation was complete within 1 hr for small ligand molecules and within 2 hr for larger ones. ...

1980-06-01

307

Curious Results with Palladium- and Platinum-Carrying Polymers in Mass Cytometry Bioassays and an Unexpected Application as a Dead Cell Stain.

The purpose was to develop a simplified and reliable method of separating free from antibody-bound ligand using a precipitating antibody linked to a cellulose derivative. Dose-response curves and control sera were set up in parallel for various pituitary and placental polypeptides, steroid hormones, insulin, glucagon, triiodothyronine, thyroxine, angiotensin I, calcitonin, gastrin, cyclic AMP, and digoxin. After first-antibody reactions had reached equilibrium, free and bound ligand were separated using a double-antibody solid-phase system in parallel with conventional methods, including dextran-coated charcoal, double-antibody precipitation, single-antibody solid phase, organic solvents, salt precipitation, and anion-exchange resins. The effect of variations in temperature, incubation time, protein content, pH, and amount of separating material added were studied. The results showed that separation was complete within 1 hr for small ligand molecules and within 2 hr for larger ones. ...

308

Clinical utility of serum ferritin levels in patients with malignant tumors treated by radiotherapy

We describe the synthesis of metal-chelating polymers (MCPs) with four different pendant polyaminocarboxylate ligands (EDTA, DTPA, TTHA, DOTA) and an orthogonal end-group, either a fluorescein molecule or a bismaleimide linker for antibody attachment. Polymer characterization by a combination of (1)H NMR, UV/vis absorption measurements, and thermal gravimetric analysis (TGA) indicated that each chain of the fluorescein-terminated polymers contained one dye molecule. These polymer samples were loaded with three different types of lanthanide ions as well as palladium and platinum ions. The numbers of metal atoms per chain were determined by a combination of UV/vis and conventional ICP-MS measurements. The experiments with lanthanide ions demonstrated that a net anionic charge on the polymer is important for water solubility. These experiments also showed that at least one type of lanthanide ion (La(3+)) is capable of forming a bimetallic complex with pendant DTPA groups. Conditions were ...

2011-09-28

309

Characterization of the human lipoprotein lipase (LPL) promoter: Evidence of two cis-regulatory regions, LP-[alpha] and LP-[beta] of importance for the differentation-linked induction of the LPL gene during adipogenesis

The serum levels of ferritin in 394 patients, including 339 patients with various malignant tumors and 23 with various non-malignant diseases, and 32 healthy subjects were determined. The normal levels of ferritin were 82.7 +- 42.3 ng/ml for males and 42.0 +- 36.9 ng/ml for females. The positive ratio of serum ferritin level was 28% in patients with malignant tumors and 22% in patients with non-malignant diseases. The usefulness of serum ferritin assay in screening for malignant tumors appeared to be limited. High serum levels of ferritin were found in patients with malignant lymphoma (positive ratio: 42%), pulmonary cancer (38%) and esophageal cancer (37%). According to the histological types, epidermoid cancer appeared to produce a higher serum level of ferritin than adenocarcinoma in patients with pulmonary cancer. It was interesting that malignant lymphoma had a high serum level of ferritin in spite of its low serum level of CEA. Carcinoma of the digestive ...

1983-10-01

310

Beneficiation-hydroretort processing of US oil shales, engineering study

When preadipocytes differentiate into adipocytes, several differentiation-linked genes are activated. Lipo-protein lipase (LPL) is one of the first genes induced during this process. To investigate early events in adipocyte development, we have focused on the transcriptional activation of the LPL gene. For this purpose, we have cloned and fused different parts of intragenic and flanking sequences with a chloramphenicol acetyltransferase reporter gene. Transient transfection experiments and DNase I hypersensitivity assays indicate that several positive as well as negative elements contribute to transcriptional regulation of the LPL gene. When reporter gene constructs were stably introduced into preadipocytes, we were able to monitor and compare the activation patterns of different promoter deletion mutants at selected time points representing the process of adipocyte development. We could delimit two cis-regulatory elements important for gradual activation of the ...

1992-10-01

311

A novel chenodeoxycholic derivative HS-1200 induces apoptosis in human HT-29 colon cancer cells

This report describes a beneficiation facility designed to process 1620 tons per day of run-of-mine Alabama oil shale containing 12.7 gallons of kerogen per ton of ore (based on Fischer Assay). The beneficiation facility will produce briquettes of oil shale concentrate containing 34.1 gallons of kerogen per ton (based on Fischer Assay). The beneficiation facility will produce briquettes of oil shale concentrate containing 34.1 gallons of kerogen per ton (based on Fischer Assay) suitable for feed to a hydroretort oil extraction facility of nominally 20,000 barrels per day capacity. The beneficiation plant design prepared includes the operations of crushing, grinding, flotation, thickening, filtering, drying, briquetting, conveying and tailings empoundment. A complete oil shale beneficiation plant is described including all anticipated ancillary facilities. For purposes of determining capital and operating costs, the ...

1988-12-01

312

In-vitro radioimmunoassay of prostate specific antigen (PSA) for the screening and management of prostate cancer in Lebanon

To investigate the growth inhibitory effects, and the underlying mechanism of human colon cancer cell (HT-29) death, induced by a new synthetic bile derivative (HS-1200). Human colon cancer cells (HT-29), in exponential growth phase, were treated with various concentrations of a new synthetic bile acid derivative (HS-1200). The growth inhibitory effects on HT-19 cells were examined using a trypan blue exclusion assay. The extent of apoptosis was determined using agarose gel electrophoresis, TUNEL assays and Hoechst staining. The apoptotic cell death was also confirmed by Western blotting of PARP, caspase-3 and DNA fragmentation factor (DFF) analysis. To investigate the involvement of mitochondria, we employed immunofluorescent staining of cytochrome c and mitochondrial membrane potential analyses. The dose required for the half maximal inhibition (IC_5_0) of the HT-29 cell growth was 100 #approx# 150 #mu# M of HS-1200. Several changes, ...

2002-12-01

313

Interaction of antimicrobial peptides with lipid membranes

Full text: Immunoassays for prostate-specific antigen (PSA) are used to detect early-stage prostate cancer, monitor disease progress, and evaluate therapeutic response. At least two forms of PSA, free PSA (F-PSA) and PSA complexed to alpha-1 anti-chymotrypsin (PSA-ACT) are detected by commercial PSA assays. The fraction of F-PSA is shown to be smaller in patients with untreated prostate cancer than in patients with benign prostate hyperplasia (BPH). Thus, combined measurements of both total and free PSA are used for a better discrimination between BPH and prostate cancer. Detection of PSA for screening of prostate cancer has been a subject of debate for many years. The reason of this debate is mainly because screening for prostate cancer is not cost-effective, as was shown by studies undertaken in Europe and United States. In Lebanon, no previous programs of screening for prostate cancer were done and so the incidence of this cancer is not known. Recently, the ...

2004-01-01

314

Uranium isotopic assay instrument

This study aims to investigate the difference in the interaction of antimicrobial peptides with two classes of zwitterionic peptides, phosphatidylethanolamines (PE) and phosphatidylcholines (PC). Further experiments were performed on model membranes prepared from specific bacterial lipids, lipopolysaccharides (LPS) isolated from Salmonella minnesota. The structure of the lipid-peptide aqueous dispersions was studied by small-and wide-angle X-ray diffraction during heating and cooling from 5 to 85 C. The lipids and peptides were mixed at lipid-to-peptide ratios 10-10000 (POPE and POPC) or 2-50 (LPS). All experiments were performed at synchrotron soft condensed matter beamline A2 in Hasylab at Desy in Hamburg, Germany. The phases were identified and the lattice parameters were calculated. Alamethicin and melittin interact in similar ways with the lipids. Pure POPC forms only lamellar phases. POPE forms lamellar phases at low temperatures that upon heating transform ...

315

Monitoring of DNA and cytogenetic damage in lymphocytes from persons with skin cancer diseases

Full text: The isotopic assay instrument under development at Pacific Northwest National Laboratory (PNNL) is capable of rapid prescreening to detect small and rare particles containing high concentrations of uranium in a heterogeneous sample. The isotopic measurement concept is based on laser vaporization of solid samples followed by sensitive isotope-specific detection using either uranium atomic fluorescence emission or uranium atomic absorbance. Both isotopes are measured concurrently, following a single ablation laser pulse using two external-cavity violet diode lasers. The simultaneous measurement of both isotopes enables the correlation of the fluorescence and absorbance signals on a shot-to-shot basis. This measurement approach demonstrated negligible channel crosstalk between isotopes. Scanning the heterogeneous samples provides high-resolution imagery of sample isotopic fluorescence and absorbance. Isotopically selective excitation of uranium vapor also ...

2006-10-16

316

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

There is a lot of interest in the studies that would help to understand whether there is a casual association between cancer and various types of molecular or cytogenetic damage detected in human cells. One major oncogenesis process is activation of proto-oncogenes by point mutations or chromosomal translocation. There are substantial evidence that indicates that the loss of heterozygosity of certain chromosomes is involved in human cancerogenesis. Our study aimed to elicit the possible association between cancer and DNA and cytogenetic abnormalities induced in lymphocytes of people bearing various categories of skin cancer cells. Fresh blood was collected by venipuncture from 25 individuals (including nine prior to cancer treatment). All patients were nonsmoking males, however 42.3 % of them were former smokers. Blood samples were divided into two parts and in the first part of samples cytogenetic studies were performed immediately, while from the second part lymphocytes were isolated ...

2004-11-01

317

[Study of body composition by potassium-40 measurement in patients with breast and uterine cancer].

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

2006-07-15

318

[Dysspermia due to inflammation. The evaluation of sperm cultures].

Total potassium was assayed in 150 normal weight and obese females (cervical carcinoma-52, endometrial carcinoma-25 and breast cancer-73) by measurements of 40K spontaneous radiation in a low-background chamber. Control group included 30 healthy and 25 obese females. Computations of body cell and extracellular mass and fat were carried out on the basis of the said measurements. Extracellular fluid volume was measured in 38 patients by X-ray fluorescence method using sodium bromide. The results pointed to a body cell mass deficit matched by increased extracellular mass due to a higher fat level in patients with breast, endometrial and cervical carcinoma. The said correlation was particularly pronounced in obese patients. The beneficial effect of treatment was more often observed in patients with normal body weight. PMID:3739295

1986-01-01

319

[Clinico-hormonal correlations in patients with chronic alcoholic intoxication].

The study evaluates 160 cases of positive spermioculture taken from 522 sterile individuals examined by the authors at the Couple Sterility Outpatient unit in Department A of the Institute of Gynecology and Obstetrics at Turin University during the period between January 1984 and December 1993. The germs responsible for infection were assayed in order to evaluate the strains which showed the highest incidence every year. Whereas there was no significant change in the absolute number of cases of sterility over the period, the number of cases caused by infection increased significantly during the second five-year period. It was found that the germs predominantly implicated in the genesis of male sterility formed part of the so-called mixed flora group, responsible in women for syndromes of often asymptomatic bacterial vaginosis which are not identified and consequently not treated. PMID:8559444

320

Zebrafish embryo extracts promote sphere-forming abilities of human melanoma cell line

Radioimmunochemical assay was used to study the hypophyseal and peripheral hormones activity in 60 patients with chronic alcoholism, stage II. A correlation has been established between the patient's age and prolactin and FSH concentrations, as well as between the duration of the recent hard drinking and the concentrations of prolactin, testosterone, FSH and interstitial cell stimulating hormone. It has been shown that the manifestation of the alcoholic abstinent syndrome depended on the prolactin concentration. The test sensitivity estimated by the prolactin level rise and the testosterone level reduction reached 92.3%. The specificity of the changes detected comprised 25%. A conclusion has been made that the disorders noted in the patients with chronic alcoholic intoxication can be used as an objective test in the alcoholism diagnosis. PMID:3936321

1985-01-01

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321

British Library Electronic Table of Contents (United Kingdom)

Sphere-forming abilities in culture condition are considered a hallmark of cancer stem-like cells, which represents tumor cell invasiveness and stem-like characteristics. We aimed to show that the sphere-forming subpopulation of human malignant melanoma cell line WM-266-4 acts differently to zebrafish embryo extracts compared with their bulk counterpart. Spheres were maintained in neural stem cell culture conditions. The embryos of zebrafish at specific developmental stages were collected and the extracts were purified under 100 kDa. Spheres were treated with embyo extracts and proliferation assay and immunocytochemistry were conducted. Spheroid cells expressed nestin and epidermal growth factor receptor (EGFR) but not melanoma antigen recognized by T-cells (MART)1, indicating their stem-l...

2009-01-01

322

Uranium Mass and Isotopic Analysis. A new drum tomographic gamma scanning system

Two dimensional NMR and NMR relaxation studies of coal structure

The NIS-6 group at Los Alamos National Laboratory pioneered the development of Tomographic Gamma Scanning and continue to improve its utility in real measurement situations. Building on this experience we report the development of a state of the art TGS system suitable for industrial applications. The image reconstruction and visualization software has been integrated into the familiar Canberra NDA2000 software suite which also handles all other aspects of the data acquisition and management. In this paper we describe the salient design and performance parameters of the system. The application of gamma-ray tomography to safeguards for improved assay accuracy and knowledge confirmation is discussed. In particular we compare and contrast the relative merits of this TGS and a traditional Segmented Gamma Scanner and illustrate how the 3-D visualization of the TGS image can provide valuable additional information.

2003-05-01

323

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

This report covers the progress made on the title project for the project period. Four major areas of inquiry are being pursued. Advanced solid state NMR methods are being developed to assay the distribution of the various important functional groups that determine the reactivity of coals. Special attention is being paid to methods that are compatible with the very high magic angle sample spinning rates needed for operation at the high magnetic field strengths available today. Polarization inversion methods utilizing the difference in heat capacities of small groups of spins are particularly promising. Methods combining proton-proton spin diffusion with [sup 13]C CPMAS readout are being developed to determine the connectivity of functional groups in coals in a high sensitivity relay type of experiment. Additional work is aimed at delineating the role of methyl group rotation in the proton NMR relaxation behavior of coals.

1992-11-25

324

The efficacy of simulated solar disinfection (SODIS) against Ascaris, Giardia, Acanthamoeba, Naegleria, Entamoeba and Cryptosporidium

Four-enzyme section of the shikimate pathway (Aro B, D, E, and K) of Streptococcus pneumoniae has been studied. Kinetic properties of the individual enzymes and three- and four-enzyme linked reactions have been characterized in vitro. On the basis of the data measured in spectrophotometric and LC-MS experiments, kinetic mechanisms of the enzymes have been suggested and all kinetic parameters have been identified. Kinetic models for these three- and four-enzyme sections of the shikimate pathway have been constructed and validated. The model of the four-enzyme section of shikimate pathway has been employed to design an inhibition-sensitive reconstituted pathway for a high-throughput screening effort on the shikimate pathway. It was demonstrated that using the model it was possible to optimize this reconstituted pathway in such a way to provide equal sensitivity of the enzymes to inhibition. PMID:16921527

2006-11-01

325

British Library Electronic Table of Contents (United Kingdom)

The antimicrobial activity of simulated solar disinfection (SODIS) in the presence and absence of riboflavin against various protozoa and helminth organisms was investigated in this study. Assays were conducted in transparent 12 well microtitre plates containing a suspension of test organisms in the presence or absence of 250mM riboflavin. Plates were exposed to simulated sunlight at an optical irradiance of 550Wm^-^2 (watts per square metre) delivered from a SUNTEST(TM) CPS+ solar simulator. Aliquots of the test suspensions were taken at set time points and the viability of the test organisms was determined by either culture, microscopy or flow cytometry where applicable. With Acanthamoeba, Naegleria, Entamoeba and Giardia exposure to SODIS at an optical irradiance of 550Wm^-^2 for up to ...

2011-01-01

326

The effects of high-intensity pulsed electromagnetic field on proliferation and differentiation of neural stem cells of neonatal rats in vitro

British Library Electronic Table of Contents (United Kingdom)

Summary The effects of high-intensity pulsed electromagnetic stimulation (HIPEMS) on proliferation and differentiation of neonatal rat neural stem cells in vitro were investigated. Neural stem cells derived from neonatal rats were exposed to 0.1 Hz, 0.5-10 Tesla (T) [8 groups of B-I, respectively], 5 stimuli of HIPEMF. The sham exposure controls were correspondingly established. Inverted phase contrast microscope was used to observe the cultured cells, MTT assay to detect the viability of the cells as expressed by absorbance (A) value, and flow cytometry to measure differentiation of neural stem cells. The results showed that A values of neural stem cells in both 3.0 T and 4.0 T groups were significantly higher than the other groups 24 to 168 h post HPEMS, indicating a strong promotion of ...

2009-01-01

327

The content of L-carnitine in meat after different methods of heat treatment

British Library Electronic Table of Contents (United Kingdom)

Purpose - The objective of this study is to examine the effects of pan-frying, boiling and cooking in a microwave on the amount of L-carnitine in meat and to look at its distribution in the surrounding fluid after food processing. Design/methodology/approach - Total carnitine, free carnitine and acylcarnitines were determined in meat samples from beef, pork and poultry (including ostrich) and in a liver sample from beef. The measurements were carried out before and after the specimens were subjected to different heat treatments. A radio-enzymatic assay was used for measurement of L-carnitine. Results are expressed per 100 gram dry matter and per 100 gram wet weight. Findings - Except for pan-frying, virtually no losses of carnitine occurred during the different procedures of heat treatment...

2011-01-01

328

The antiviral action of common household disinfectants and antiseptics against murine hepatitis virus, a potential surrogate for SARS coronavirus

British Library Electronic Table of Contents (United Kingdom)

Background The 2003 outbreak of severe acute respiratory syndrome (SARS) infected over 8000 people and killed 774. Transmission of SARS occurred through direct and indirect contact and large droplet nuclei. The World Health Organization recommended the use of household disinfectants, which have not been previously tested against SARS coronavirus (SARS-CoV), to disinfect potentially contaminated environmental surfaces. There is a need for a surrogate test system given the limited availability of the SARS-CoV for testing and biosafety requirements necessary to safely handle it. In this study, the antiviral activity of standard household products was assayed against murine hepatitis virus (MHV), as a potential surrogate for SARS-CoV. Methods A surface test method, which involves drying an amo...

2009-01-01

329

Syringe calibration factors and volume correction factors for the NPL secondary standard radionuclide calibrator

CERN Document Server

The activity assay of a radiopharmaceutical administration to a patient is normally achieved via the use of a radionuclide calibrator. Because of the different geometries and elemental compositions between plastic syringes and glass vials, the calibration factors for syringes may well be significantly different from those for the glass containers. The magnitude of these differences depends on the energies of the emitted photons. For some radionuclides variations have been observed of 70 %, it is therefore important to recalibrate for syringes or use syringe calibration factors. Calibration factors and volume correction factors have been derived for the NPL secondary standard radionuclide calibrator, for a variety of commonly used syringes and needles, for the most commonly used medical radionuclide.

2002-01-01

330

Symbiotic N2 fixation by legumes growing in pots. 2. Uptake of VN-labelled NO3 , C2H2 reduction and H2 evolution by Trifolium subterraneum L. , Medicago truncatula Gaertn. and Acacia dealbata Link

Summary of reference materials for the determination of the nutrient composition of foods

The objectives of this study were to estimate symbiotic nitrogen fixation by two common pasture legumes, Trifolium subterraneum L. and Medicago truncatula Gaertn., and an Australian native legume, Acacia dealbata Link, growing in pots using an indirect isotopic method. This method was also used to calibrate the C2H2 reduction assay of the intact plants. In addition, hydrogen evolution was measured in an attempt to explain the variations in C2H2:N2 ratios between the species. 25 refs.; 1 figure; 4 tabs.

1983-01-01

331

British Library Electronic Table of Contents (United Kingdom)

Certified reference materials (CRMs) play a critical role in validating the accuracy of nutrient data for food samples. A number of available food CRMs of differing matrix composition have assigned concentrations for various nutrients, along with associated uncertainty intervals (UIs) for those values. These CRMs have been used extensively in the United States Department of Agriculture?s (USDA) ongoing National Food and Nutrient Analysis Program (NFNAP) to monitor the accuracy of assays of key foods and nutrients consumed in the United States. A total of 690 assigned values for individual nutrients, including proximates, vitamins, macroelements, microelements, fatty acids, amino acids, and selected phytochemicals (e.g., carotenoids), were compiled from the certificates of analysis for 63 C...

2007-01-01

332

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

Studies of the fundamental nature of catalytic acidity, sites, and intermediates: Progress report for period August 1, 1987-July 31, 1988

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

333

Structure-activity relationships of anthraquinone derivatives derived from bromaminic acid as inhibitors of ectonucleoside triphosphate diphosphohydrolases (E-NTPDases)

Neopentane, because of its unique structural features, was found to be a useful probe for the assay of the acidity of various zeolites. The C-C bond was attacked by catalyst protons yielding CH/sub 4/ and the isobutyl carbenium ion in equal quantities. The latter either decomposed to produce isobutene or underwent secondary reactions. New avenues for reaction were opened with isobutane. The reaction of the Bronsted protons with the tertiary CH bonds produced H/sub 2/ and the same isobutyl carbenium ion. Hydride ion transfer of the same tertiary hydrogen to existing carbenium ions also occurred and the results showed that about half of the reaction was carried by this pathway. As with Neopentane, the C-C bond could also be attacked yielding the sec-propyl carbenium ion. These tools are being further developed.

1988-06-01

334

British Library Electronic Table of Contents (United Kingdom)

Reactive blue 2 (RB-2) had been characterized as a relatively potent ectonucleoside triphosphate diphosphohydrolase (E-NTPDase) inhibitor with some selectivity for NTPDase3. In search for the pharmacophore and to analyze structure-activity relationships we synthesized a series of truncated derivatives and analogs of RB-2, including 1-amino-2-sulfo-4-ar(alk)ylaminoanthraquinones, 1-amino-2-methyl-4-arylaminoanthraquinones, 1-amino-4-bromoanthraquinone 2-sulfonic acid esters and sulfonamides, and bis-(1-amino-4-bromoanthraquinone) sulfonamides, and investigated them in preparations of rat NTPDase1, 2, and 3 using a capillary electrophoresis assay. Several 1-amino-2-sulfo-4-ar(alk)ylaminoanthraquinone derivatives inhibited E-NTPDases in a concentration-dependent manner. The 2-sulfonate group ...

2009-01-01

335

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

Rapid determination of granisetron in human plasma by liquid chromatography coupled to tandem mass spectrometry and its application to bioequivalence study

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

1992-03-01

336

British Library Electronic Table of Contents (United Kingdom)

A simple, sensitive and rapid method for analysis of granisetron in human plasma, utilizing liquid chromatography tandem mass spectrometry (LC-MS/MS), has been developed and validated to satisfy FDA guidelines for bioanalytical methods. The analyte and internal standard (IS) were isolated from 100ml plasma samples by liquid-liquid extraction (LLE). A Varian 1200l tandem mass spectrometer equipped with an electrospray ionization source was operated in selected reaction monitoring (SRM) mode with the precursor-to-product ion transitions m/z 313.4/138 for granisetron and m/z 270/201 for the IS used for quantitation. The assay exhibited a linear dynamic range of 0.02-20ng/ml for granisetron in human plasma. The lower limit of quantification (LLOQ) was 0.02ng/ml with a relative standard deviati...

2006-01-01

337

Rapamycin (sirolimus) protects against hypoxic damage in primary heart cultures via Na^+/Ca^2^+ exchanger activation

British Library Electronic Table of Contents (United Kingdom)

Aims: Rapamycin (sirolimus) is an antibiotic that inhibits protein synthesis through mammalian targeting of rapamycin (mTOR) signaling, and is used as an immunosuppressant in the treatment of organ rejection in transplant recipients. Rapamycin confers preconditioning-like protection against ischemic-reperfusion injury in isolated mouse heart cultures. Our aim was to further define the role of rapamycin in intracellular Ca^2^+ homeostasis and to investigate the mechanism by which rapamycin protects cardiomyocytes from hypoxic damage. Main methods: We demonstrate here that rapamycin protects rat heart cultures from hypoxic-reoxygenation (H/R) damage, as revealed by assays of lactate dehydrogenase (LDH) and creatine kinase (CK) leakage to the medium, by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-d...

2011-01-01

338

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

British Library Electronic Table of Contents (United Kingdom)

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

2011-01-01

339

Radioimmunoassay data processing, ch. 2

Proteomic analysis of apoptosis induction in human lung cancer cells by recombinant MVL

The problems of radioimmunoassay data processing with a fairly big computer are investigated in detail. The logit transformation, introduced by Rodbard et al. in 1968, and the related mathematical operations are presented. From logit-ln plots the confidence limits for the calculation of the antigen concentration can be predicted as well as the minimum detectable dose (i.e. the sensitivity of the assay). The Scatchard plot is used to calculate the equilibrium constant of the antigen-antibody coupling reaction and the binding capacity. A 4-parameter logistic curve fitting method is developed and combined with a method to carry out a multi-component Scatchard analysis. Both a weighted and an unweighted linear regression procedure are evaluated for use in calculating the plasma renin activity from the angiotensin I amounts determined by radioimmunoassay. Improvements for Rodbard's computer programmes are proposed.

1978-02-15

340

British Library Electronic Table of Contents (United Kingdom)

Lung cancer is still difficult to treat by current chemotherapeutic procedures. We recently found that MVL, an anti-HIV lectin from blue-green algae Microcystis viridis, also has antitumor activity. The objective of this study was to investigate apoptosis-inducing activity of recombinant MVL (R-MVL) and proteomic changes in A549 cells, and to identify the molecular pathways responsible for the anti-cancer action of R-MVL. We found that R-MVL induces A549 cells apoptosis in a dose-dependent manner by using MTT assay, fluorescent microscope (FM) and flow cytometry (FCM), and the IC50 was calculated to be 24.12??g/ml. Subsequently, 7 altered proteins in R-MVL-treated A549 cells were identified, including upregulated aldehyde dehydrogenase 1 and ?-actin, and five downregulated proteins: heat s...

2011-01-01

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341

Preliminary studies on the chemical characterization and antioxidant properties of acidic polysaccharides from Sargassum fusiforme

British Library Electronic Table of Contents (United Kingdom)

In order to investigate the antioxidant properties of the polysaccharides from the brown alga Sargassum fusiforme, the crude polysaccharides from S. fusiforme (SFPS) were extracted in hot water, and the lipid peroxidation inhibition assay exhibited that SFPS possessed a potential antioxidant activity. Hence, two purely polymeric fractions, SFPS-1 and SFPS-2 were isolated by the column of DEAE (2-diethylaminoethanol)-Sepharose Fast Flow, with their molecular weights of 51.4 and 30.3 kDa determined by high performance gel permeation chromatography (HPGPC). They were preliminarily characterized using chemical analysis in combination of infrared (IR) and nuclear magnetic resonance (NMR) spectroscopies and found to contain large amounts of uronic acids and ?-glycosidical linkages. The antioxida...

2008-01-01

342

Potential Protection of Green Tea Polyphenols Against 1800?MHz Electromagnetic Radiation-Induced Injury on Rat Cortical Neurons

British Library Electronic Table of Contents (United Kingdom)

Radiofrequency electromagnetic fields (EMF) are harmful to public health, but the certain anti-irradiation mechanism is not clear yet. The present study was performed to investigate the possible protective effects of green tea polyphenols against electromagnetic radiation-induced injury in the cultured rat cortical neurons. In this study, green tea polyphenols were used in the cultured cortical neurons exposed to 1800?MHz EMFs by the mobile phone. We found that the mobile phone irradiation for 24?h induced marked neuronal cell death in the MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-tetrazolium bromide) and TUNEL (TdT mediated biotin-dUTP nicked-end labeling) assay, and protective effects of green tea polyphenols on the injured cortical neurons were demonstrated by testing the content ...

2011-01-01

343

Pectinolytic yeast isolates for cold-active polygalacturonase production

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

344

Paraquat toxicity is increased in Escherichia coli defective in the synthesis of polyamines

PREPARATION AND PROPERTIES OF ANTIBACTERIAL ALGINATE FILMS INCORPORATING EXTRUDED WHITE GINSENG EXTRACT

The authors have shown that toxicity of paraquat for Escherichia coli is increased over 1-fold in strains defective in the biosynthesis of spermidine compared to isogenic strains containing spermidine. The increased sensitivity of these spermidine-deficient mutants to paraquat is eliminated by growth in medium containing spermidine or by endogenous supplementation of spermidine by the use of a speE"+D"+ plasmid. No paraquat toxicity is seen in the absence of oxygen, even in amine-deficient strains, indicating that superoxide is the agent responsible for the increased toxicity. However, the specific mechanisms responsible for the increased paraquat toxicity in the spermidine-deficient mutants remain to be determined. The marked sensitivity to paraquat of E. coli deficient in spermidine is of particular interest, since such mutants have no other phenotypic properties that can be easily assayed. This increased sensitivity has been used as the basis of a convenient ...

345

British Library Electronic Table of Contents (United Kingdom)

Abstract Preparation of antibacterial alginate films incorporating extruded white ginseng (EWG) extracts was attempted. The antibacterial effect of EWG extract on six selected food pathogenic bacteria was compared with the effect of red ginseng (RG) and white ginseng (WG) extracts. The EWG was processed in a twin-screw extruder with feed moisture of 20% and barrel temperature of 115 and 130C. The data obtained by agar diffusion assay demonstrated that the film containing 1-g/mL of EWG at barrel temperatures of 115 (EWG-115) and 130C (EWG-130) exhibited stronger antibacterial activity against the four strains of bacteria than the other extracts RG and WG. All films sampled showed reduction in bacteria cell counts (log cycle) compared with the control. After 24-h of incubation, Pseudomonas a...

2011-01-01

346

Oral administration of purple passion fruit peel extract attenuates blood pressure in female spontaneously hypertensive rats and humans

British Library Electronic Table of Contents (United Kingdom)

Hypertension is one of the most important modifiable risk factors for cardiovascular and cerebrovascular diseases. We investigated the potential antihypertensive effect of the purple passion fruit peel (PFP) extract, a mixture of bioflavonoids, phenolic acids, and anthocyanins, in spontaneously hypertensive rats and human. A high-performance liquid chromatography analysis was performed to identify the active ingredients of the PFP extract. In a rat liver toxicity assay, no hepatotoxicity was observed after 9 hours incubation in the presence of PFP extract (20 ?g/mL). The PFP extract also revealed hepatoprotection against chloroform (1 mmol/L)-induced liver injury. In the experimental model, 24 spontaneously hypertensive rats were divided into 3 treatment groups for a period of 8weeks: cont...

2007-01-01

347

Nondestructive characterization of low-level transuranic waste

No detectable XMRV in subjects with chronic fatigue syndrome from Quebec

The use of nondestructive evaluation (NDE) methods is proposed for characterization of transuranic (TRU) waste stored at the Radioactive Waste Management Complex. These NDE methods include real-time x-ray radiography, real-time neutron radiography, x-ray and neutron computed tomography, thermal imaging, container weighing, visual examination, and acoustic measurements. An integrated NDE system is proposed for characterization and certification of TRU waste destined for eventual shipment to the Waste Isolation Pilot Plant in New Mexico. Methods for automating both the classification waste and control of a complete nondestructive evaluation/nondestructive assay system are presented. Feasibility testing of the different NDE methods, including real-time x-ray radiography, and development of automated waste classification techniques are covered as part of a five year effort designed to yield a production waste characterization system.

1981-10-01

348

British Library Electronic Table of Contents (United Kingdom)

We investigated the presence of XMRV in a cohort of Quebec patients with chronic fatigue syndrome (CFS). DNA was purified from activated peripheral blood mononuclear cells (PBMCs) and PCR was used to detect XMRV gag and env in 72 patients. Anti-XMRV antibodies were searched in sera of 62 patients by Western blot analysis. Attempts to detect XMRV antigens was made, using immunofluorescence with Gag anti-p30 antiserum on activated PBMC from 50 patients. Plasma viremia was measured by RT-PCR on 9 subjects. Finally, detection of infectious virus in 113 CFS subjects was made by co-culture of PHA+IL-2 activated PBMC with human LNCaP carcinoma cells, and by infecting the same susceptible cells with plasma, using a reverse transcriptase (RT) assay as a readout in both experiments. No detection of ...

2011-01-01

349

Mutations at the cysteine codons of the recA gene of Escherichia coli

Involvement of a putative response regulator Brrg-1 in the regulation of sporulation, sensitivity to fungicides, and osmotic stress in Botrytis cinerea

Each of the three cysteine residues in the Escherichia coli RecA protein was replaced with a number of other amino acids. To do this, each cysteine codon was first converted to a chain-terminating amber codon by oligonucleotide-directed mutagenesis. These amber mutants were then either assayed for function in different suppressor strains or reverted by a second round of mutagenesis with oligonucleotides that had random sequences at the amber codon. Thirty-three different amino acid substitutions were obtained. Mutants were tested for three functions of RecA: survival following UV irradiation, homologous recombination, and induction of the SOS response. It was found that although none of the cysteines is essential for activity, mutations at each of these positions can affect one or more of the activities of RecA, depending on the particular amino acid substitution. In addition, the cysteine at position 116 appears to be involved in the RecA-promoted cleavage of the ...

350

British Library Electronic Table of Contents (United Kingdom)

The response regulator protein is a core element of two-component signaling pathway. In this study, we investigated functions of BRRG-1 of Botrytis cinerea, a gene that encodes a putative response regulator protein, which is homologous to Rrg-1 in Neurospora crassa. The BRRG-1 gene deletion mutant ?Brrg1-62 was unable to produce conidia. The mutant showed increased sensitivity to osmotic stress mediated by NaCl and KCl, and to oxidative stress generated by H2O2. Additionally, the mutant was more sensitive to the fungicides iprodione, fludioxonil, and triadimefon than the parental strain. Western-blot analysis showed that the Bos-2 protein, the putative downstream component of Brrg-1, was not phosphorylated in the ?Brrg1-62. Real-time polymerase chain reaction assays showed that expression ...

2011-01-01

351

Investigation of Tumor-Peritoneal Interactions in the Pathogenesis of Peritoneal Metastases using a Novel Ex Vivo Peritoneal Model1

British Library Electronic Table of Contents (United Kingdom)

BackgroundPeritoneal metastasis occurs in up to 30% of patients with gastric cancer. The aim of this experimental study is to develop and validate a novel ex vivo model of the human peritoneum to better identify factors involved in the development of peritoneal metastasis in order to improve its management and prognosis. MethodsPeritoneal discs harvested from hernia sacs obtained at inguinal hernia surgery were suspended in media using Teflon rings. Viability of the tissue was investigated using MTS assay, light and scanning electron microscopy (LM and SEM) over 72 h. To assess validity of the model, phenotypic changes in tumor cells were investigated. Changes in matrix metalloproteinases (MMP)-2 and -9 activities in HGC and AGS gastric adenocarcinoma cells after co-culture were investigat...

2010-01-01

352

Inhibitory activity of Pseudomonas sp. on Flavobacterium psychrophilum, in vitro

British Library Electronic Table of Contents (United Kingdom)

Abstract A Pseudomonas sp. isolate MSB1 efficiently inhibited the growth of Flavobacterium psychrophilum of different serotypes on agar medium. A significant difference in the inhibition was observed between isolates of the less virulent FpT serotype compared to the Fd and Th serotypes. In broth coculture experiments, a low number of cells of MSB1 inhibited and outcompeted the F.psychrophilum cells. Also cell-free culture supernatant of MSB1 clearly repressed the growth of F.psychrophilum. A chromoazurol S assay suggested that MSB1 produced efficient siderophores, which most probably were responsible for the iron deficiency in the supernatant. The limited growth of F.psychrophilum in the supernatant was found to be partly because of the lack of available iron, but the results also indicate...

2011-01-01

353

In vitro and in vivo evaluation of [{sup 123}I]-VEGF{sub 165} as a potential tumor marker

Hsp90 gene, an additional target for discrimination between the potato cyst nematodes, Globodera rostochiensis and G. pallida, and the related species, G. tabacum tabacum

One of the research challenges in oncology is to develop new biochemical methods for noninvasive tumor therapy evaluation to determine whether the chemotherapeutics is effective. Vascular endothelial growth factor (VEGF) was labeled with radioiodine and evaluated in vitro as well as in vivo, using A2058, a melanoma cell line overexpressing VEGFR-1 and -2. Saturation binding analysis with [{sup 125}I]-VEGF resulted in a K {sub d} of 0.1 nM. Internalization assays indicate the preserved ligand induced internalization and metabolization of the tracer. Biodistribution studies with [{sup 123}I]-VEGF in wild type and A2058 tumor-bearing athymic mice showed low background activity and a tumor to reference tissue ratio of maximum 6.12. These results suggest that [{sup 123}I]-VEGF is a potentially suitable tracer for tumor therapy evaluation.

2005-07-01

354

British Library Electronic Table of Contents (United Kingdom)

The heat-shock gene, Hsp90, was targeted as a new variable genomic region to supplement other DNA-based tests for identification and discrimination of Globodera pallida, G. rostochiensis and G. tabacum tabacum. Populations of the potato cyst nematodes, G. pallida and G. rostochiensis (PCN), originating from Canada, France, Belgium and USA, together with two populations of G. tabacum tabacum from the USA and France were used for the amplification of a fragment of the Hsp90 gene. General and specific primers and probes for each species were derived from the consensus and non-consensus regions of the aligned sequences, respectively. A triplex conventional PCR assay, using a general forward and reverse or three specific reverse primers, as well as a real-time PCR using general primers and spec...

2011-01-01

355

Homologous radioimmunoassay for human prolactin

High resolution gamma-ray spectrometry of culverts containing transuranic waste at the Savannah River Site

Although thee are descriptions of a range of radioimmunoassays for human prolactin in various biological fluids, only one of these is an homologous assay using human prolactin as the reference standard and tracer as well and an anti-human prolactin antiserum (Sinha, Y.N., Selby, F.W.; Lewis, U.; and Vanderlaan, W.P., 1973, J. Clin. Endocr., Vol. 36, 509). A homologous radioimmunoassay using human putuitary prolactin has been developed. The separation method is based on the double antibody solid phase system. Cross reactivity with human growth hormone (GH), placental lactogen (HPL), the pituitary protein hormones and prolactins of various species were studied as were values found in normal subjects in basal conditions and after a TRH injection. (author).

356

High resolution gamma-ray spectrometry of culverts containing transuranic waste at the Savannah River Site

A number of concrete culverts used to retrievably store drummed, dry, radioactive waste at the Savannah River Site (SRS), were suspected of containing ambiguous quantities of transuranic (TRU) nuclides. These culverts were assayed in place for Pu-239 content using thermal and fast neutron counting techniques. High resolution gamma-ray spectroscopy on 17 culverts, having neutron emission rates several times higher than expected, showed characteristic gamma-ray signatures of neutron emitters other than Pu-239 (e.g., Pu-238, Pu/Be, or Am/Be neutron sources). This study confirmed the Pu-239 content of the culverts with anomalous neutron rates and established limits on the Pu-239 mass in each of the 17 suspect culverts by in-field, non-intrusive gamma-ray measurements.

1990-01-01

357

Functional domain analysis of glass, a zinc-finger-containing transcription factor in Drosophila.

A number of concrete culverts used to retrievably store drummed, dry, radioactive waste at the Savannah River Site (SRS), were suspected of containing ambiguous quantities of transuranic (TRU) nuclides. These culverts were assayed in place for Pu-239 content using thermal and fast neutron counting techniques. High resolution gamma-ray spectroscopy on 17 culverts, having neutron emission rates several times higher than expected, showed characteristic gamma-ray signatures of neutron emitters other than Pu-239 (e.g., Pu-238, Pu/Be, or Am/Be neutron sources). This study confirmed the Pu-239 content of the culverts with anomalous neutron rates and established limits on the Pu-239 mass in each of the 17 suspect culverts by in-field, non-intrusive gamma-ray measurements.

1990-12-31

358

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

The glass gene is required for proper photo-receptor differentiation during development of the Drosophila eye glass codes for a DNA-binding protein containing five zinc fingers that we show is a transcriptional activator. A comparison of the sequences of the glass genes from two species of Drosophila and a detailed functional domain analysis of the Drosophila melanogaster glass gene reveal that both the DNA-binding domain and the transcriptional-activation domain are highly conserved between the two species. Analysis of the DNA-binding domain of glass indicates that the three carboxyl-terminal zinc fingers alone are necessary and sufficient for DNA binding. We also show that a deletion mutant of glass containing only the DNA-binding domain can behave in a dominant-negative manner both in vivo and in a cell culture assay that measures transcriptional activation. PMID:7604032

1995-07-01

359

Expression of Distal-less, dachshund, and optomotor blind in Neanthes arenaceodentata (Annelida, Nereididae) does not support homology of appendage-forming mechanisms across the Bilateria

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

2007-07-01

360

British Library Electronic Table of Contents (United Kingdom)

The similarity in the genetic regulation of arthropod and vertebrate appendage formation has been interpreted as the product of a plesiomorphic gene network that was primitively involved in bilaterian appendage development and co-opted to build appendages (in modern phyla) that are not historically related as structures. Data from lophotrochozoans are needed to clarify the pervasiveness of plesiomorphic appendage-forming mechanisms. We assayed the expression of three arthropod and vertebrate limb gene orthologs, Distal-less (Dll), dachshund (dac), and optomotor blind (omb), in direct-developing juveniles of the polychaete Neanthes arenaceodentata. Parapodial Dll expression marks pre-morphogenetic notopodia and neuropodia, becoming restricted to the bases of notopodial cirri and to ventral ...

2010-01-01

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

361

Enhanced osteogenic differentiation of cord blood-derived unrestricted somatic stem cells on electrospun nanofibers

British Library Electronic Table of Contents (United Kingdom)

A new stem cell-scaffold construct based on poly-l-lactide (PLLA) nanofibers grafted with collagen (PLLA-COL) and cord blood-derived unrestricted somatic stem cells (USSC) were proposed to hold promising characteristics for bone tissue engineering. Fabricated nanofibers were characterized using SEM, ATR-FTIR, tensile and contact angle measurements. The capacity of PLLA, plasma-treated PLLA (PLLA-pl) and PLLA-COL scaffolds to support proliferation and osteogenic differentiation of USSC was evaluated using MTT assay and common osteogenic markers such as alkaline phosphatase (ALP) activity, calcium mineral deposition and bone-related genes. All three scaffolds showed nanofibrous and porous structure with suitable physical characteristics. Higher proliferation and viability of USSC was observe...

2011-01-01

362

Development of radioimmunometric assays and kits for non-clinical applications. Proceedings of a final research coordination meeting

DNFB-DNS hapten-induced colitis in mice should not be considered a model of inflammatory bowel disease

Immunoassays are versatile analytical techniques that had a leading role in various clinical applications, during the last four decades. The studies carried out by Rosalyn Yalow, Solomon Berson and Roger Ekins in the 1960s gave a breakthrough in the development of this novel analytical method. Sensitivity up to femtomolar concentrations, high specificity and universal application to different classes of molecular species made immunoassay a very useful tool in analytical investigation. The expertise acquired by immunochemists in producing antibodies against any antigen and the ability of radiochemists in labeling the antigens with "1"2"5I without affecting the active site are the two main factors responsible for the above development. There are concerns about the safety and health of humans due to the high-level contamination of environment by pesticides, industrial compounds and metals, anabolic steroids in milk and meat products, and presence of mycotoxins in food and feed ...

2004-12-06

363

British Library Electronic Table of Contents (United Kingdom)

AbstractBackground: The dinitrofluorobenzene/dinitrosulfonic acid (DNFB/DNS) model was originally described as an experimental model of intestinal inflammation resembling human ulcerative colitis (UC). Due to the absence of acceptable UC experimental models for pharmacological preclinical assays, here we examine the immune response induced in this model. Methods: Balb/c mice were sensitized by skin application of DNFB on day 1, followed by an intrarectal challenge with DNS on day 5. We further expanded this model by administering a second DNS challenge on day 15. The features of colonic inflammation and immune response were evaluated. Results: The changes observed in colonic tissue corresponded, in comparison to the trinitrobenzene sulfonic acid (TNBS) colitis model, to a mild mucosal effe...

2011-01-01

364

Control of late blight in organic potato production: evaluation of copper-free preparations under field, growth chamber and laboratory conditions

British Library Electronic Table of Contents (United Kingdom)

In order to replace copper fungicides in organic potato production, 53 copper-free preparations (CFPs) based on natural compounds, including plant extracts and microorganisms, and five copper preparations were evaluated for their potential to control Phytophthora infestans, the pathogen that causes late blight of potatoes. In in vitro assays, 30% of the CFPs inhibited indirect germination of sporangia, 26% mycelial growth and in growth chamber experiments, 21% efficiently reduced foliar blight of tomato plants. In micro-plot field trials with applications twice a week, the copper preparations were the most effective and reduced foliar blight by 99%. Of the CFPs tested, Oekofluid P, Mycosin and other sulphuric clays, and C-2000 reduced late blight the most, from 63% to 37%. In small-plot tr...

2007-01-01

365

Comparison of the results obtained by CALUX bioassay and GC-HRMS for different matrices

Comparative biochemistry of betaine biosynthesis and accumulation in diverse dicot families

The reference method used to analyse polychlorodibenzo-p-dioxins (PCDDs), polychlorodibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dl-PCBs) is chromatography with high resolution mass spectrometry (GC-HRMS). It is interesting to check the suitability of screening methods that are faster and less expensive. Different matrices (milk, fish oil, chicken compound feed, pork tissue, chicken tissue, sepiolitic clay, whole egg and herring tissue) were analysed in the frame of the European project DIFFERENCE1. One of the aims of this project is to optimise screening methods. The CALUX bio-assay was one of the screening techniques used. This paper presents the extraction and purification methods used for the analyses. The CALUX results for dioxins and for dl-PCBs were compared to the corresponding GC-HRMS results.

2004-09-15

366

Chemical Analysis of Solid-State Irradiated Human Insulin

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

1989-04-01

367

British Library Electronic Table of Contents (United Kingdom)

Purpose To study the chemical modifications induced upon irradiation of solid human insulin at radiosterilization doses and investigate the influence of the absorbed dose on radiolysis. Materials and Methods Volatile radiolytic products were monitored by gas chromatography coupled with mass spectrometry (GC-MS) and non-volatile products by two different high performance liquid chromatography (HPLC) methods: the formation of higher molecular weight proteins was assessed by size exclusion liquid chromatography whereas assays for related compounds and chemical potency tests were carried out using reverse-phase HPLC-UV. Conformational changes were investigated by measurements of circular dichroism. Results After gamma irradiation at 10?kGy, the recovery of insulin was 96.8%; higher molecular w...

2006-01-01

368

Bacterial adhesion reduction on a biocompatible Si^+ ion implanted austenitic stainless steel

British Library Electronic Table of Contents (United Kingdom)

The colonization of an implant surface by bacteria is an extremely important medical problem, which often leads to the failure of medical devices. Modern surface modification techniques, such as ion implantation, can confer to the surfaces very different properties from those of the bulk underlying material. In this work, austenitic stainless steel 316 LVM has been superficially modified by Si^+ ion implantation. The effect of surface modification on the biocompatibility and bacterial adhesion to 316 LVM stainless steel has been investigated. To this aim, human mesenchymal stem cells (hMSCs), as precursor of osteoblastic cells, and bacterial strains relevant in infections related to orthopedic implants, i.e., Staphylococcus aureus and Staphylococcus epidermidis, have been assayed. For the ...

2011-01-01

369

Avian gut-passage effects on seed germination of shrubland species in Mediterranean central Chile

British Library Electronic Table of Contents (United Kingdom)

Effects of avian gut-passage on seed germination are important to assess the effectiveness of frugivores in woodland regeneration, particularly in biodiversity hotspots that have a high incidence of avian frugivory. We examined the effect of avian gut-passage on seed germination in contrast to seeds that remain uneaten in five shrub species in Mediterranean central Chile and sought to determine the physiological mechanism(s) by which seed germinability is modified. Germination assays were conducted in a glasshouse for five common shrub species of the sub-Andean matorral: Azara dentata (Flacourtiaceae), Schinus polygamus and Schinus molle (Anacardiaceae), Cestrum parqui (Solanaceae), and Maytenus boaria (Celastraceae). We estimated germinability (final percent germination), dormancy length ...

2011-01-01

370

Association between chemical and genetic variation in Calophyllum inophyllum, a medicinally important tree of the Western Ghats of India

British Library Electronic Table of Contents (United Kingdom)

The objective of the present work is to study the chemical variation in Calophyllum inophyllum growing along the Western Ghats of India. Contents of dipyranocoumarins (inophyllums) in C.?inophyllum were determined to assess whether they could be used as a taxonomic marker for C.?inophyllum. This study also aims to establish inter simple sequence repeat (ISSR) markers that can be used to study genetic variation within the species and explore correlation between ISSR and chemical markers. The contents of dipyranocoumarins were estimated in seeds collected from 20 locations. Leaves from plants at the same 20 locations were assayed for ISSR variation. A dendrogram based on Nei?s genetic distance as well as principal component analysis based on dipyranocoumarins and ISSR variation clustered pla...

2011-01-01

371

Assessment of different protocols for the isolation and purification of gut associated lymphoid cells from the gilthead seabream (Sparus aurata L.)

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...

2007-01-01

372

Antioxidant phytochemicals and gingerol content in diploid and tetraploid clones of ginger (Zingiber officinale Roscoe)

British Library Electronic Table of Contents (United Kingdom)

Fresh rhizomes of 18 diploid and tetraploid genotypes of Indian ginger, including commercial cultivars and experimental genotypes, were assayed for gingerol content and antioxidant activity. The tetraploid ginger type was derived from the respective diploid ginger by shoot tip culture. [6]-Gingerol was identified as the major pungent phenolic compound in all samples (mean 1354.78+/-470mg/g), while [8]- (mean 180.94+/-45mg/g) and [10]-gingerol (mean 234+/-79mg/g) occurred in lower concentrations. The total gingerol content of the tetraploid type was much higher than that of the respective diploid type and especially the [10]-gingerol. Both diploid and tetraploid types of the cv. Meghalaya Local contained the highest level of gingerol. At both ploidy levels, the genotypes showed significant ...

2010-01-01

373

Antioxidant activity in banana peel extracts: Testing extraction conditions and related bioactive compounds

British Library Electronic Table of Contents (United Kingdom)

Banana (Musa acuminata Colla AAA) peel extracts obtained in this work had a high capacity to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2prime-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS+) free radicals, and they were also good lipid peroxidation inhibitors. Acetone:water extracts were considerably more effective (compared with methanol, ethanol, acetone, water, methanol:water or ethanol:water) at inhibiting the peroxidation of lipids in the b-carotene/linoleic acid system or scavenging free radicals. However, aqueous extracts had a high capacity to protect lipids from oxidation in the thiobarbituric acid reactive substances (TBARS) test, as well as in the b-carotene bleaching assay. In addition, acetone:water most efficiently extracted all extractable components (54+-...

2010-01-01

374

Analysis and assay of flavonoid content in the vegetative and reproductive organs of the showy tick trefoil

British Library Electronic Table of Contents (United Kingdom)

We report here a many-year study of the qualitative and quantitative content of flavonoids in the reproductive organs of the showy tick trefoil (Desmodium canadense (L.) DC., Fabaceae) - stems, leaves, buds, inflorescences,pods, and seeds - collected at different phases of growth at the Department of Medicinal Plants, Kaunas Botanical Garden, Vytautas Magnus University. HPLC studies identified 15 aglycone and glycoside flavonoids: apigenin, apigenin-7-O-glycoside, luteolin, rutin, vicenin-2, vitexin, isovitexin, vitexin rhamnoside, orientin, homoorientin, quercetrin, quercetin, hyperoside, astralagin, and kaempferol. The largest quantities of flavonoids in the vegetative organs from year 2 to year 4 of plant growth were present in the leaves (2.64% and 2.61% at budding and flowering in the...

2011-01-01

375

An erythrocyte-specific DNA-binding factor recognizes a regulatory sequence common to all chicken globin genes

An application of UV-derivative spectrophotometry and bivariate calibration algorithm for study of photostability of levomepromazine hydrochloride

The authors have identified a protein present only in erythroid cells that binds to two adjacent sites within an enhancer region of the chicken {beta}-globin locus. Mutation of the sites, so that binding by the factor can no longer be detected in vitro, leads to a loss of enhancing ability, assayed by transient expression in primary erythrocytes. Binding sites for the erythroid-specific factor (Eryf1) are found within regulatory regions for all chicken globin genes. A strong Eryf1 binding site is also present within the enhancer of at least one human globin gene, and proteins from human erythroid cells (but not HeLa cells) bind to both the chicken and the human sites.

1988-08-01

376

British Library Electronic Table of Contents (United Kingdom)

Derivative spectrophotometry and bivariate calibration algorithm were used for study of run of photooxidation of levomepromazine hydrochloride (LV). The actual concentrations of LV and its main degradation product levomepromazine sulphoxide (LV-SO) were calculated using data provided by applied methods. The direct reading of absorbance values at 302nm and 334nm were employed for quantification of LV and LV-SO, respectively, in the case of bivariate method. The derivative spectrophotometric method is based on transformation of zero-order spectra into first derivative. The values of first derivative at 334nm were used for quantification of LV while at 278nm for assay of LV-SO. The obtained quantitative data were applied for investigation of kinetics of photodegradation of LV.

2008-01-01

377

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

British Library Electronic Table of Contents (United Kingdom)

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

2008-01-01

378

Activation of rapid signaling pathways and the subsequent transcriptional regulation for the proliferation of breast cancer MCF-7 cells by the treatment with an extract of Glycyrrhiza glabra root

British Library Electronic Table of Contents (United Kingdom)

Glycyrrhiza glabra root is one of the common traditional Chinese medicines and used as flavoring and sweetening agents for tobaccos, chewing gums, candies, toothpaste and beverages. While glycyrrhizin is one of the main components in the extract of G. glabra root and has been characterized, the other components have not been well characterized. The mechanism of growth activation of breast cancer MCF-7 cells, including the activation of Erk1/2 and Akt, and the transcriptional regulation of estrogen-responsive genes, was examined by means of sulforhodamine B, luciferase reporter gene, real-time RT-PCR and Western blotting assays after the induction of the cells with the extract of G. glabra root. The extract has similar activity to that induced by 17b-estradiol (E2), although glycyrrhizin di...

2007-01-01

379

A variable gene delivery carrier-biotinylated chitosan/polyethyleneimine

A radioassay for Gsub(M1) ganglioside concentration in cerebrospinal fluid

A variable gene delivery system has been developed based on conjugating chitosan to biotin through a functionalized poly(ethylene glycol) (PEG) spacer, which can be used to further bind different molecules on the outer layer of a polymer/DNA complex by streptavidin (SA)-biotin linkage. In this study, TAT-conjugated SA was used as the model molecule to prove the conjugation function of the prepared complex. In addition, low-molecular-weight poly(ethyleneimine) (PEI) was added into the polymer/DNA complex to increase the transfection efficiency. The results of the luciferase assay show that the transfection efficiency of the prepared complex was significantly correlated with the amount of PEI and was further enhanced when TAT was conjugated to the complex by SA-biotin linkage. Considered to have negligible cytotoxic effects, the variable gene delivery complex prepared in this study would be of considerable potential as carriers for in vitro applications.

2010-12-01

380

A protocol for the evaluation of genotoxicity in bile of carp (Cyprinus carpio) exposed to lake water treated with different disinfectants

A radioassay for the rapid determination of Gsub(M1) ganglioside concentration in small volumes of CSF from individual patients is described. The assay utilizes the high-affinity interaction between cholera enterotoxin ("1"2"5I-labelled) and Gsub(M1) ganglioside. The lower detection limit of Gsub(M1) ganglioside by this radioassay under the described incubation conditions is 2.5 ng/ml. The radioassay-determined lumbar CSF Gsub(M1) ganglioside concentrations in a small group of patients with diverse neurologic disorders are presented. The radioassay Gsub(M1) ganglioside concentration is in good agreement with the G(M1) ganglioside concentration determined, in one patient, by the tlc-densitometry technique. (author).

1980-01-01

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381

British Library Electronic Table of Contents (United Kingdom)

A sensitive and rapid method to evaluate toxic and genotoxic properties of drinking water supplied from Lake Trasimeno (Umbria, Central Italy) was worked out analysing bile in Cyprinus carpio exposed for 20d to lake water treated with 3 different disinfectants, sodium hypochlorite (NaClO), chlorine dioxide (ClO2) and peracetic acid (PAA). Fish were sacrificed at 0, 10 and 20d in order to investigate the time course of these endpoints. An aliquot of bile samples was fractionated by adsorption on C18 silica cartridges and the genotoxic potential of whole bile and of bile fractions was evaluated by the single-cell microgel-electrophoresis (comet) assay on human colonic adenocarcinoma cells (Caco-2). Bile (both whole and fractionated) from specimens exposed to the three disinfectants always sh...

2011-01-01

382

Transcriptional Profiling Analysis of the Global Regulator NorG, a GntR-Like Protein of Staphylococcus aureus.

Transcription regulation of the vegf gene by the BMP/Smad pathway in the angioblast of zebrafish embryos

The GntR-like protein NorG has been shown to affect Staphylococcus aureus genes involved in the resistance to quinolones and ?-lactams, such as those encoding the NorB and AbcA transporters. To identify the target genes regulated by NorG, we carried out transcriptional profiling assays using S. aureus RN6390 and its isogenic norG::cat mutant. Our data showed that NorG positively affected the transcription of global regulators mgrA, arlS, and sarZ. The three putative drug efflux pump genes most positively affected by NorG were the NorB efflux pump (5.1-fold), the MmpL-like protein SACOL2566 (5.2-fold), and the BcrA-like drug transporter SACOL2525 (5.7-fold). The S. aureus predicted MmpL protein showed 53% homology with the MmpL lipid transporter of Mycobacterium tuberculosis, and the putative SACOL2525 protein showed 87% homology with the bacitracin drug transporter BcrA of Staphylococcus hominis. Two pump genes most negatively affected by NorG were NorC (4-fold) ...

2011-09-01

383

Toxicological safety evaluation of biomolecules and materials transformed by gamma irradiation

Vascular endothelial growth factor (VEGF) is a mitogen that is critically involved in vasculogenesis, angiogenesis, and hematopoiesis. However, what and how transcription factors participate in the regulation of vegf gene expression are not fully understood. Here we report the cloning and sequencing of the zebrafish vegf promoter which revealed that the promoter contains a number of bone morphogenetic protein (BMP)-activated Smad binding elements (SBE), implicating Smad1 and Smad5 in the regulation of BMP-induced expression of vegf. Electrophoretic mobility shift assays of adding recombinant Smad proteins to the SBE-containing DNA oligonucleotides that represent portions of zebrafish vegf promoter resulted in mobility shift of the oligonucleotides. These changes demonstrate potential interactions between Smad1/5 and the vegf promoter. Reporter activity assays using the wild-type or SBE-deleted vegf promoters to drive the luciferase reporter ...

2005-04-01

384

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

In the bacterial reversion assay with S. typhimurium TA98, TA100, TA1535 and TA1537, gamma irradiated hyaluronic acid (10 and 50 kGy) did not induce a significant increase in the number of revertant colonies in the presence of S9 metabolic activation system. In chromosomal aberration tests with CHO cells, gamma irradiated hyaluronic acid (10 and 50 kGy) did not result in an increase in the frequency of chromosomal aberrations. In vivo mouse micronucleus assay, gamma irradiated hyaluronic acid (10 and 50 kGy) did not show an increase in the frequency of polychromatic erythrocytes with micronuclei. These results indicate that hyaluronic acids irradiated at 10 and 50 kGy did not show any genotoxic effects under these experimental conditions. In order to evaluate their possible subacute toxicity, the male and female of ICR mouse were given to methanol extract of 50 kGy irradiated red ginseng and 20 kGy irradiated water extract of mistletoe for ...

2010-01-15

385

Studies on a Factor in Sweet Potato Root Which Agglutinates Spores of Ceratocystis fimbriata, Black Rot Fungus 1

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

2004-09-15

386

Status of the WAND (Waste Assay for Nonradioactive Disposal) project as of July 1997

A factor which agglutinated the spores of Ceratocystis fimbriata in the presence of Ca2+ was purified from sweet potato (Ipomea batatas Lam cv. Norin[1]) root. Element composition of the purified factor was as follows; analysis found: C (29.8%), H (3.97%), O (65.34%), N (0.81%): calculated for C43H69O70N1: C (30.02%), H (4.01%), O (65.15%), N (0.81%). The factor was mainly composed of galacturonic acid (53% of dry weight) and contained arabinose, fucose, and unidentified component as minor components. The factor also agglutinated A-, B-, AB-, and O types of human erythrocytes to almost the same degree in the presence of Ca2+. The differential spore-agglutinating activity of the factor depended on the pH of the assay medium; it agglutinated similarly the germinated spores of sweet potato and coffee strains at pH 7.5 and 5.5, whereas it displayed a distinct differential agglutinating activity at pH 6.5. The factor was assayed for ...

1982-02-01

387

Sensitivity and accuracy considerations for neutron assay of plutonium-contaminated waste in large containers

The WAND (Waste Assay for Nonradioactive Disposal) system can scan thought-to-be-clean, low-density waste (mostly paper and plastics) to determine whether the levels of any contaminant radioactivity are low enough to justify their disposal in normal public landfills or similar facilities. Such a screening would allow probably at least half of the large volume of low-density waste now buried at high cost in LANL`s Rad Waste Landfill (Area G at Technical Area 54) to be disposed of elsewhere at a much lower cost. The WAND System consists of a well-shielded bank of six 5-in.-diam. phoswich scintillation detectors; a mechanical conveyor system that carries a 12-in.-wide layer of either shredded material or packets of paper sheets beneath the bank of detectors; the electronics needed to process the outputs of the detectors; and a small computer to control the whole system and to perform the data analysis. WAND system minimum detectable activities (MDAs) for point sources ...

1998-03-01

388

Semiquantitative measure of immune responses against erythropoietic stem cell antigens

Since the 1970 innovations have allowed both active and passive neutron techniques to address various safeguards and waste measurement needs in the DOE complex. Much research was focused on satisfjring the 100-nCi/g detection limit for TRU waste in 208-liter drums. The emphasis on measuring drum-sized containers for disposal at WIPP has resulted in improved waste assay capability that now needs to be extended to larger containers. The desire to expedite the decontamination and decommissioning of certtain DOE facilities, and the large waste encountered in that process, has prompted the need for increasingly large disposal containers. Instruments have recently been built to accommodate crates that are nearly 100 cubic feet in volume, such as a B-25 box or Standard Waste 13ox. The density of hydrogen inside a waste container profoundly affects the accuracy of neutron measurements, and the metal content greatly affects sensitivity. Depending on the matrix, and ...

2001-01-01

389

Screening for congenital hypothyroidism in Lebanon

A semiquantitative assay was developed and used to measure the effects of immune responses against 16 independent non-H-2 antigenic loci on erythropoietic stem cells. The assay compares repopulation in genetically anemic WBB6F1-W/Wv recipients that have normal immune responses, and in lethally irradiated WBB6F1 +/+ mice whose immune responses are suppressed by the irradiation. The differences in repopulating ability between these two types of recipients measure how immune responses affect erythropoietic stem cells. Stem cell repopulating abilities for the cells with antigens specified by the Thy-1, H-1, H-24, Ly-1, H-37, and H-17 loci were affected slightly, if at all. Repopulating abilities were moderately reduced by responses against antigens specified by H-15, 16, Ea-2, and Ly-2, 3 loci, and against the differences between the B6 and B10 genotypes, although marrow of these types cured W/Wv recipients. A surprising result occurred for the ...

1987-01-01

390

Radiation-induced damage to DNA; Les lesions radio-induites de l'ADN

Full text.National programs of neonatal screening were introduced in most countries and have proved their effectiveness against different types of retardation in children. Congenital hypothyroidism is one of the most common causes of physical and mental retardation in children. Programs of screening have shown that the incidence of this disorder is ethnic-background: it is low in Africans: 1-32000 whereas it is 1-3500 and 1-5000 in Hispanic and European newborns respectively. No previous mass screening programs were done in Lebanon. For this purpose, this program was introduced in collaboration with the IAEA. Cord blood is spotted into circles on filter paper just after delivery. These samples are air-dried at room temperature and kept at 4 degree C. They are then collected from hospitals to be screened. In the laboratory, all samples are assayed for IRMA TSH first. Those showing high or borderline TSH level (20mU/L) are then assayed for RIA ...

1999-11-02

391

Oleuropein aglycon prevents cytotoxic amyloid aggregation of human amylin.

This short survey focuses on the main radiation-induced base lesions that have been identified within cellular DNA. For this purpose, sensitive assays that are aimed at measuring a few modifications per 10{sup 7} normal bases were set-up. In that respect high performance liquid chromatography - tandem mass spectrometry (CLHP-MS/MS) was found to be able to single out the formation of 9 oxidized nucleosides and two modified nucleo-bases out of the 70 oxidative base lesions that have been identified in model systems. As a striking result, it was found that in the DNA of {gamma}-irradiated human monocytes, the formamide-pyrimidine derivative of guanine is produced in a higher yield than the ubiquitous 8-oxo-7,8-dihydro-guanine damage, both arising from the same radical precursor. However, relatively high doses of ionizing radiation (> 20 Gy) have to be applied in order to detect an increase in the level of the damage. This is due to the low efficiency for both ...

2002-03-01

392

In vivo studies of the long-term "5"1Cr red cell survival of serologically incompatible red cell units

Pancreatic amyloid deposits of amylin are a hallmark of Type II diabetes and considerable evidence indicates that amylin oligomers are cytotoxic to beta-cells. Many efforts are presently spent to find out naturally occurring molecules, or to design synthetic ones, able to hinder amylin aggregation or to protect cells against aggregate cytotoxicity. In this context, a protective effect of some polyphenols against amyloid cytotoxicity was reported. Actually dietary polyphenols are endowed with multiple health benefits, and extra virgin olive oil is attracting increasing interest as a source of these substances. Here, we investigated the effects on amylin aggregation and cytotoxicity of the secoiridoid oleuropein aglycon, the main phenolic component of extra virgin olive oil. We found that oleuropein, when present during the aggregation of amylin, consistently prevented its cytotoxicity to RIN-5F pancreatic beta-cells, as determined by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl ...

2009-07-18

393

In vitro comparison of rat and chicken brain neurotoxic esterase

The long-term survival of serologically incompatible red cell units was measured in five patients with antibodies to high-frequency antigens. Initially, the survival of 1 ml of "5"1Cr-labeled incompatible red cells was measured over 1 hour. After demonstrating that the 1-hour survival times were successful (greater than 70%), each patient then received 5 ml of the same "5"1Cr-labeled red cells followed by the transfusion of the remainder of the red cell unit. The long-term T 1/2Cr survival for each case was patient 1 (anti-McCa), 15 days; patient 2 (anti-JMH), 12 days; patient 3 (anti-Kna), 31 days; patient 4 (anti-McCa), 12 days; and patient 5 (anti-Hya), 14 days. Each antibody tested in an in vitro homologous macrophage assay showed less than 5 percent phagocytosis. Anti-JMH was the only antibody to react with IgG subclass antisera and was determined to be IgG4. The macrophage assay, IgG subclass testing, and short-term (1 hour, 1 ml) "5"1Cr ...

394

Genotoxicity of 2-alkylcyclobutanones, markers for an irradiation treatment in fat-containing food--Part I: cyto- and genotoxic potential of 2-tetradecylcyclobutanone

A systematic comparison was undertaken to characterize neurotoxic esterase (NTE) from rat and chicken brain in terms of inhibitor sensitivities, pH optima, and molecular weights. Paraoxon titration of phenyl valerate (PV)-hydrolyzing carboxylesterases showed that rat esterases were more sensitive than chicken to paraoxon inhibition at concentrations less than or equal to microM and superimposable with chicken esterases at concentrations of 2.5-1000 microM. Mipafox titration of the paraoxon-resistant esterases at a fixed paraoxon concentration of 100 microM (mipafox concentration: 0-1000 microM) resulted in a mipafox I50 of 7.3 microM for chicken brain NTE and 11.6 microM for rat brain NTE. NTE (i.e., paraoxon-resistant, mipafox-sensitive esterase activity) comprised 80% of chicken and 60% of rat brain paraoxon-resistant activity with the specific activity of chicken brain NTE approximately twice that of rat brain NTE. The pH maxima for NTE from both species was similar showing broad, ...

1986-04-01

395

Functional and physical molecular size of the chicken hepatic lectin determined by radiation inactivation and sedimentation equilibrium analysis

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. Neither cytotoxic nor genotoxic effects ...

2002-03-01

396

ERK-dependent and -independent pathways trigger human neural progenitor cell migration

Radiation inactivation and sedimentation equilibrium analysis were used to determine the functional and physical size of the chicken hepatic membrane receptor that binds N-acetylglucosamine-terminated glycoproteins. Purified plasma membranes from chicken liver were irradiated with high energy electrons and assayed for 125I-agalactoorosomucoid binding. Increasing the dose of ionizing radiation resulted in a monoexponential decay in binding activity due to a progressive loss of binding sites. The molecular mass of the chicken lectin, determined in situ by target analysis, was 69,000 +/- 9,000 Da. When the same irradiated membranes were solubilized in Brij 58 and assayed, the binding protein exhibited a target size of 62,000 +/- 4,000 Da; in Triton X-100, the functional size of the receptor was 85,000 +/- 10,000 Da. Sedimentation equilibrium measurements of the purified binding protein yielded a lower limit molecular weight of 79,000 +/- 7,000. ...

1990-03-05

397

Development of radiological emergency preparedness technology

Besides differentiation and apoptosis, cell migration is a basic process in brain development in which neural cells migrate several centimeters within the developing brain before reaching their proper positions and forming the right connections. For identifying signaling events that control neural migration and are therefore potential targets of chemicals to disturb normal brain development, we developed a human neurosphere-based migration assay based on normal human neural progenitor (NHNP) cells, in which the distance is measured that cells wander over time. Applying this assay, we investigated the role of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) in the regulation of NHNP cell migration. Exposure to model substances like ethanol or phorbol 12-myristate 13-acetate (PMA) revealed a correlation between ERK1/2 activation and cell migration. The participation of phospho-(P-) ERK1/2 was confirmed by exposure of the cells to the ...

2007-05-15

398

Dendroaspis natriuretic peptide and the designer natriuretic peptide, CD-NP, are resistant to proteolytic inactivation.

Large-scale field tracer experiments have been conducted on Ulchin, Wolsung and Daeduk sites for the purpose of validating FADAS and of analyzing the environmental characteristics around the nuclear sites. The most influential factor in atmospheric dispersion is the meteorological condition. During the experiment, meteorological data were measured on the release point and the selected positions among sampling points. Once radioactive materials are released to the atmosphere, members of public may be exposed through the environmental media such as air, soil and foods. Therefore, to protect the public, adequate countermeasures should be taken at due time for those exposure pathways. both processes, of justification and optimization are applied to a countermeasure simultaneously for decision-making. The work scope of Biological research for the radiation protection had contained the search of biological microanalytic methods for assessing the health effect by {gamma}-radiation and toxic ...

2000-04-01

399

Characterization and geochemistry of Devonian oil shale: north Alabama, northwest Georgia, and south-central Tennessee (a resource evaluation). Final report

Designer natriuretic peptides (NPs) represent an active area of drug development. In canine and human studies, the designer natriuretic peptide CD-NP demonstrated more desirable therapeutic potential than recombinant B-type NP (BNP), which is known as nesiritide and is approved for treatment of acute decompensated heart failure. However, why CD-NP is more effective than BNP is not known. We previously reported that CD-NP is a poorer activator of human guanylyl cyclase-A (GC-A) and a better activator of human guanylyl cyclase-B than BNP. Here, guanylyl cyclase bioassays were used to compare the susceptibility of CD-NP verses ANP, BNP, CNP and DNP to inactivation by human kidney membranes. The half time (t(1/2)) for CD-NP inactivation was increased by factors of 13, 3 and 4 compared to ANP, BNP and CNP, respectively, when measured in the same assay. Surprisingly, DNP failed to undergo complete inactivation and was the most degradation resistant of the peptides ...

2011-04-01

400

An in vitro model to test relative antioxidant potential: Ultraviolet-induced lipid peroxidation in liposomes

A geochemical survey to assess and evaluate the Devonian oil shale resources of north Alabama, northwest Georgia, and south-central Tennessee has been performed. Information is provided on the location and extent of the more favorable kerogen-bearing Devonian shale, oil and uranium content of the shale, trace metal and standard rock analyses for shale, data on the physical properties of the shale rock, mineralogy of the shale, chemical properties of the oil, and other pertinent data deemed necessary for resource development. The results indicate that the oil yield from areas underlain by Chattanooga Shale can be divided into areas having greater than 7 g/t and areas having less than 7 g/t. In general, the oil yield by Fischer assay of samples from the folded Appalachian Valley and Ridge and the Appalachian Plateaus provinces is poor, all having less than 5 g/t and most samples closer to 0 g/t. The extreme variance of Fischer assay values in the ...

1984-05-03

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401

Activation of rapid signaling pathways and the subsequent transcriptional regulation for the proliferation of breast cancer MCF-7 cells by the treatment with an extract of Glycyrrhiza glabra root.

Since antioxidants have been shown to play a major role in preventing some of the effects of aging and photoaging in skin, it is important to study this phenomenon in a controlled manner. This was accomplished by developing a simple and reliable in vitro technique to assay antioxidant efficacy. Inhibition of peroxidation by antioxidants was used as a measure of relative antioxidant potential. Liposomes, high in polyunsaturated fatty acids (PUFA), were dispersed in buffer and irradiated with ultraviolet (UV) light. Irradiated liposomes exhibited a significantly higher amount of hydroperoxides than liposomes containing antioxidants in a dose- and concentration-dependent manner. Lipid peroxidation was determined spectrophotometrically by an increase in thiobarbituric acid reacting substances. To further substantiate the production of lipid peroxides, gas chromatography was used to measure a decrease in PUFA substrate. In order of decreasing antioxidant effectiveness, ...

402

A survey of monitoring and assay systems for release of metals from radiation controlled areas at LANL.

Glycyrrhiza glabra root is one of the common traditional Chinese medicines and used as flavoring and sweetening agents for tobaccos, chewing gums, candies, toothpaste and beverages. While glycyrrhizin is one of the main components in the extract of G. glabra root and has been characterized, the other components have not been well characterized. The mechanism of growth activation of breast cancer MCF-7 cells, including the activation of Erk1/2 and Akt, and the transcriptional regulation of estrogen-responsive genes, was examined by means of sulforhodamine B, luciferase reporter gene, real-time RT-PCR and Western blotting assays after the induction of the cells with the extract of G. glabra root. The extract has similar activity to that induced by 17beta-estradiol (E(2)), although glycyrrhizin did not show such an activity. Moreover, the estrogen receptor alpha-dependent neurite outgrowth induced by the extract was similar to that by E(2), whereas glycyrrhizin had no ...

2007-06-24

403

Two dimensional NMR and NMR relaxation studies of coal structure. Progress report, July 1, 1992--September 30, 1992

At Los Alamos National Laboratory (LANL), a recent effort in waste minimization has focused on scrap metal from radiological controlled areas (RCAs). In particular, scrap metal from RCAs needs to be dispositioned in a reasonable and cost effective manner. Recycling of DOE scrap metals from RCAs is currently under a self-imposed moratorium. Since recycling is not available and reuse is difficult, often metal waste from RCAs, which could otherwise be recycled, is disposed of as low-level waste. Estimates at LANL put the cost of low-level waste disposal at $550 to $4000 per cubic meter, depending on the type of waste and the disposal site. If the waste is mixed, the cost for treatment and disposal can be as high as $50,000 per cubic meter. Disposal of scrap metal as low-level waste uses up valuable space in the low-level waste disposal areas and requires transportation to the disposal site under Department of Transportation (DOT) regulations for low-level waste. In contrast, disposal as ...

2002-01-01

404

Total phenolics and antioxidant activity of five medicinal plant; Fenois totais e atividade antioxidante de cinco plantas medicinais

This report covers the progress made on the title project for the project period. Four major areas of inquiry are being pursued. Advanced solid state NMR methods are being developed to assay the distribution of the various important functional groups that determine the reactivity of coals. Special attention is being paid to methods that are compatible with the very high magic angle sample spinning rates needed for operation at the high magnetic field strengths available today. Polarization inversion methods utilizing the difference in heat capacities of small groups of spins are particularly promising. Methods combining proton-proton spin diffusion with {sup 13}C CPMAS readout are being developed to determine the connectivity of functional groups in coals in a high sensitivity relay type of experiment. Additional work is aimed at delineating the role of methyl group rotation in the proton NMR relaxation behavior of coals.

1992-11-25

405

The growth factor from plerocercoids of Spirometra mansonoides is both a growth hormone agonist and a cysteine proteinase.

This paper describes total phenolics content and antioxidant activity in the ethanolic extract of leaves, bark and roots of five medicinal plants: Terminalia brasiliensis Camb., Terminalia fagifolia Mart. and Zucc., Copernicia cerifera (Miller) H.E. Moore, Cenostigma macrophyllum Tul. var. acuminata Teles Freire and Qualea grandiflora Mart. The total phenolics content of the plant extracts, determined by the Folin-Ciocalteu method, varied from 250.0 {+-}8,2 to 763,63 {+-}13.03 mg of gallic acid equivalent/g dry EtOH extract. The antioxidant activity of extracts was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay system. Extract of bark from T. brasiliensis, the most active, with an EC{sub 50} value of 27.59 {+-} 0.82 {mu}g/mL, was comparable to rutin (EC{sub 50} = 27.80 {+-} 1.38) and gallic acid (EC{sub 50} = 24.27 {+-} 0.31), used as positive controls. The relationship between total phenolic content and antioxidant activity was positive and ...

2007-03-15

406

The effects of high-intensity pulsed electromagnetic field on proliferation and differentiation of neural stem cells of neonatal rats in vitro.

Plerocercoids of the tapeworm Spirometra mansonoides produce a substance that stimulates growth of experimental hosts. We report purification of plerocercoid growth factor (PGF) to homogeneity by a process involving isolation and solubilization of plerocercoid membranes, isoelectric point selection by chromatofocusing chromatography or preparative isoelectric focusing, and anion-exchange chromatography. A radioreceptor assay (RRA) for human growth hormone (hGH) was used to detect PGF and purity of the 27.5-kDa protein was judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Proteolytic activity was detected in the 27.5-kDa protein by gelatin substrate PAGE. Characterization of PGF as a neutral cysteine proteinase was based on substrate and inhibitor specificities and dependence on pH and thiol-containing reagents. The association of hGH agonist and proteinase activities was shown by comparing RRA and hydrolytic activities in the presence ...

1996-04-01

407

The development of sigma-receptor specific radiopharmaceuticals for spect imaging of human tumors

The effects of high-intensity pulsed electromagnetic stimulation (HIPEMS) on proliferation and differentiation of neonatal rat neural stem cells in vitro were investigated. Neural stem cells derived from neonatal rats were exposed to 0.1 Hz, 0.5-10 Tesla (T) [8 groups of B-I, respectively], 5 stimuli of HIPEMF. The sham exposure controls were correspondingly established. Inverted phase contrast microscope was used to observe the cultured cells, MTT assay to detect the viability of the cells as expressed by absorbance (A) value, and flow cytometry to measure differentiation of neural stem cells. The results showed that A values of neural stem cells in both 3.0 T and 4.0 T groups were significantly higher than the other groups 24 to 168 h post HPEMS, indicating a strong promotion of the growth of neural stem cells (PHPEMS groups was the same as that in control group (P>0.05). It was suggested that 0.1 Hz, 5 pulses stimulation of HPEMS within certain scale of ...

2009-12-29

408

Temporal development of GABA agonist induced alterations in ultrastructure and GABA receptor expression in cultured cerebellar granule cells

The authors have been involved for the last few years in developing high affinity sigma-receptor-specific radiopharmaceuticals for use in nuclear medicine. They recently demonstrated that sigma-1 receptors are expressed in human melanoma cells and that sigma receptors could be used as external markers for non-invasive imaging. In order to determine the presence of sigma receptors in other human tumors they have screened numerous non-small cell lung carcinoma, melanoma and breast cancer cell lines. All of these cell lines expressed high sigma receptor densities using commercially available ["3H]-(+)-pentazocine or ["3H]DTG tritiated sigma ligands. Therefore, a sigma ligand with high affinity in both sigma-1 and sigma-2 subtype pharmacological assays could be potentially useful for imaging a variety of primary and metastatic sites of various origins. The authors describe recent efforts to develop high affinity sigma radiopharmaceuticals and pharmacological studies to ...

1994-08-21

409

DEFF Research Database (Denmark)

The temporal development of the effect of THIP (4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol) on the ultrastructure composition and GABA receptor expression in cerebellar granule cells was investigated by quantitative electron microscopy (morphometric analysis) and GABA binding assays. It was found that the cytoplasmic density of smooth endoplasmic reticulum was decreased, while the cytoplasmic density of rough endoplasmic reticulum, Golgi apparatus, vesicles and coated vesicles was greatly enhanced after exposure of the cells to THIP (150 microM) for only 1 hr. In cerebellar granule cells exposed to THIP (150 microM) for 3 hr low affinity GABA receptors were induced. These findings show that the effect of THIP on the ultrastructure composition and GABA receptor expression in cultured cerebellar granule cells may be interrelated and moreover it is likely that the turn-over of GABA receptors is extremely fast.

1987-01-01

410

T-2 mycotoxin inhibits mitochondrial protein synthesis

Stem cell kinetics in spleen and bone marrow after single and fractionated irradiation of infant mice

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

411

Some thoughts on stem cells and carcinogenesis. The thyroid gland

The number and type of stem cells in spleen and bone marrow of mice were determined after exposure to a single dose of 150 R on day 6, to a single dose of 500 R on day 6 or day 9 or to a fractionated dose of 150 R + 350 R on day 6 and 9. The stem cells were assayed on the basis of colony forming units (CFU) in spleen and of incorporation of iododeoxyuridine in spleen and bone marrow of lethally irradiated host mice. During the first month of life, the number of stem cells in non-irradiated mice increases markedly in bone marrow and slightly in spleen. Irradiation causes a long-lasting depression in stem cells, particularly in bone marrow and affecting preferentially erythropoietic precursor cells. Following a dose of only 150 R, the number of CFU in bone marrow is still below control levels 24 days later. An exposure to 500 R fractionated between day 6 and 9 has a markedly greater effect on stem cells in the spleen than the same dose given in a single application ...

1980-12-01

412

Simultaneous determination of dorzolamide HCL and timolol maleate in eye drops by two different spectroscopic methods.

The purpose of this review is to consider the hypothesis that cancer frequently originates from stem cells. Using the spleen transplantation assay where stem cells were transplanted in the spleen of mice lethally irradiated by ionizing radiation, the author undertook a study aimed at defining the risk of radiogenic cancer per susceptible cells with use of rat radiogenic mammary and thyroid cancers because of the high incidences of these cancers in a-bomb survivors. Measured were the number of cancer-susceptible cells initially present in the tissue, the number of such cells that survived at a given dose and the number of cancers that developed per surviving cell. Thyroid cell differentiation and proliferation in rats transplanted with thyroid cells were enhanced by thyroidectomy and low iodine diet. Further, the relationship between the low LET radiation dose and thyroid clonogen survival was also investigated. Data showed that follicular-unit-forming clonogens ...

2000-07-01

413

Silver nanoparticles directly formed on natural macroporous matrix and their anti-microbial activities

Two-component mixtures of dorzolamide hydrochloride and timolol maleate were assayed by first derivative and ratio derivative spectrophotometric methods. The first method, derivative spectrophotometry, by the zero-crossing measurements, was used due to the drugs closely overlapping absorption spectra. Linear calibration graphs of first derivative values at 250.3 nm for dorzolamide hydrochloride and 315.8 nm for timolol maleate. The second method, is based on ratio first derivative spectrophotometry, the amplitudes in the first derivative of the ratio spectra at 242.9 and at 223.5 nm were selected to determine dorzolamide and timolol maleate in the binary mixture. Calibration graphs were established for 8.0-30.0 microg ml(-1) for dorzolamide hydrochloride and 3.0-24.6 microg ml(-1) for timolol maleate in binary mixture. Good linearity, precision and selectivity were found, and the proposed methods were applied successfully to the pharmaceutical dosage from ...

2002-04-15

414

Salivary cortisol assessment in the evaluation of hypothalamic-pituitary-adrenal function

In this study, silver nanoparticles were formed on a natural macroporous matrix, the stem of rice-paper plant, by reducing Ag{sup +} in aqueous solution through in situ processing without using any other stabilizers. The pores of the matrix, with their size of about 100 {mu}m, were thought to act as reaction compartments for the nucleation and growth of silver nanoparticles, and the control of nucleation of silver crystal during the reduction reaction was found to be important to the successful formation of nanosized silver particles onto the matrix. The diameter and amount of resultant silver particles can be controlled by changing the reaction conditions. Under optimized conditions, the content of silver particles in the matrix can reach as high as 1.8 wt% with the particle diameters being kept below 100 nm. The anti-microbial activities in terms of minimum inhibitory concentration (MIC) for the silver nanoparticle composites against Escherichia coli and Candida albicans were ...

2007-02-07

415

Regulatory role of neuron-restrictive silencing factor in expression of TRPC1

A simple, short and sensitive direct radio-immunoassay technique for the determination of salivary cortisol concentration was employed to assess saliva as a medium for evaluating cortisol response during endocrine testing in 9 controls and 40 patients. Results in controls suggested that an adequate salivary cortisol response to insulin hypoglycaemia was an increase of 150% above the basal value with a minimum peak of 15 nmol/l. Thirty-three patients were classified as being either good or poor responders to insulin hypoglycaemia on the basis of criteria for plasma cortisol levels. When the defined salivary cortisol response was used for assessment, all 33 patients were correctly categorized into the same response groups. The salivary cortisol response to intramuscular tetracosactin in 3 patients and an intravenous dexamethasone infusion in 4 patients confirmed the value of saliva as an assay medium. These studies show that the salivary cortisol response parallels ...

1984-05-01

416

Regulation of macrophage accessory cell activity by mycobacteria. I. Ia expression in normal and irradiated mice infected with Mycobacterium mycroti

Neuron-restrictive silencer factor (NRSF) binds its consensus element to repress the transcription of various genes. The dominant-negative form (dnNRSF) has a hypertrophic effect on cardiogenesis through an unidentified mechanism. We examined the involvement of transient receptor potential (TRP) channel proteins, using transgenic mice overexpressing dnNRSF (dnNRSF mice). Electrophoretic mobility-shift assays revealed an interaction between NRSF and a neuron-restrictive silencer element-like sequence in intron 4 of TRPC1 genomic DNA. According to RT-PCR and Western analyses, TRPC1 was up-regulated in dnNRSF mouse heart. Transient overexpression of TRPC1 in HEK 293T cells increased the activity of the nuclear factor in activated T cells (NFAT) promoter and stimulated store-operated Ca"2"+ channel (SOCC)-mediated Ca"2"+ entry. Transfection of TRPC1 into primary cardiomyocytes increased NFAT activity, indicating a major role for TRPC1 in NFAT activation. Our findings ...

2006-12-22

417

Reclaiming silver from silver zeolite

CBA/Ca mice were infected by either the intravenous or intraperitoneal route with Mycobacterium microti and the subsequent changes in local macrophage populations examined. Following infection, the number of macrophages increased and they showed greater expression of both MHC Class II molecules. This response was not dependent on viability of the mycobacteria, in contrast to reports with other microorganisms such as Listeria. Studies in sublethally irradiated mice indicated that persistent antigen could give rise to a response after a period of host recovery which was radiation dose dependent. This procedure also highlighted differences in the regulation of different murine class II antigens in vivo, as seen by delayed re-expression of I-E antigens. Macrophage accessory cell function, as assessed by an in vitro T cell proliferation assay, correlated with Ia expression after fixation, but not after indomethacin treatment; this highlights the diverse nature of ...

418

Reactive oxygen species production and Bax/Bcl-2 regulation in honokiol-induced apoptosis in human hepatocellular carcinoma SMMC-7721 cells.

Silver zeolite is used to capture radioiodines from air cleaning systems in some nuclear facilities at the Idaho National Engineering Laboratory. It may become radioactively contaminated and/or poisoned by hydrocarbon vapors, which diminishes its capacity for iodine. Silver zeolite contains up to 38 wt% silver. A pyrometallurgical process was developed to reclaim the silver before disposing of the unserviceable zeolite as a radioactive waste. A flux was formulated to convert the refractory aluminosilicate zeolite structure into a low-melting fluid slag, with Na{sub 2}O added as NAOH instead of Na{sub 2}CO{sub 3} to avoid severe foaming due to CO{sub 2} evolution. A propane-fired furnace was built to smelt 45 kg charges at 1300C in a carbon-bonded silicon carbide crucible. A total of 218 kg (7000 tr oz) of silver was reclaimed from 1050 kg of unserviceable zeolite. Silver recoveries of 97% were achieved, and the radioisotopes were fixed as stable silicates in a vitreous slag that was ...

1991-10-01

419

Probiotics inhibit enteropathogenic E. coli adherence in vitro by inducing intestinal mucin gene expression.

We investigated possible mechanism(s) where honokiol induces apoptosis in human hepatocellular carcinoma SMMC-7721 cells. MTT assay showed that honokiol has strong inhibition on SMMC-7721 cells in a dose-dependent manner. SMMC-7721 cells after honokiol treatment display morphological characteristics such as cell shrinkage, detachment from the culture plate, formation of apoptotic bodies, change to a round shape, and marked nuclear condensation and fragmentation after 32258 staining. Cell apoptosis was measured by Annexin-V/PI staining and alternatively, by the subG0/G1 percentage of the cell cycle analysis followed by FACS. An obvious loss of ??(m) and a quick burst of ROS was detected when honokiol reached 4?g/ml, which was coincident with the high apoptosis percentage in our previous research. Up-regulation of Bax and down-regulation of Bcl-2 were observed, suggesting that honokiol-induced apoptosis was associated with reactive oxygen species (ROS) production and ...

2009-03-17

420

Primary productivity C-14 and algal assay in the study of water pollution effects in the Citarum River and Jatiluhur Reservoir

Probiotic agents, live microorganisms with beneficial effects for the host, may offer an alternative to conventional antimicrobials in the treatment and prevention of enteric infections. The probiotic agents Lactobacillus plantarum 299v and Lactobacillus rhamnosus GG quantitatively inhibited the adherence of an attaching and effacing pathogenic Escherichia coli to HT-29 intestinal epithelial cells but did not inhibit adherence to nonintestinal HEp-2 cells. HT-29 cells were grown under conditions that induced high levels of either MUC2 or MUC3 mRNA, but HEp-2 cells expressed only minimal levels of MUC2 and no MUC3 mRNA. Media enriched for MUC2 and MUC3 mucin were added exogenously to binding assays and were shown to be capable of inhibiting enteropathogen adherence to HEp-2 cells. Incubation of L. plantarum 299v with HT-29 cells increased MUC2 and MUC3 mRNA expression levels. From these in vitro studies, we propose the hypothesis that the ability of probiotic agents ...

1999-04-01

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

421

Prediction of Skin Sensitization with a Particle Swarm Optimized Support Vector Machine

Water quality of the Citarum River and the Jatiluhur Reservoir in Indonesia was evaluated using C-14 radioisotope. A close relationship between the abiotic (physical and chemical) and the biotic (algal growth potential, primary productivity, chlorophylla and diversity index of planktonic and benthic macroinvertebrate) parameters was obtained. Algal growth potential to abiotic parameters relationship has the highest correlation coefficient and can be used as a pollution indicator. The other biotic parameters do not show clear relationship with the abiotic parameters. The Citarum water quality is the lowest in those locations which receive human and industrial waste from Bandung and its environment. This water cannot be used for drinking purposes and fishery. The water quality in other locations of the river, however, meets the criteria for agriculture. Agricultural waste does not show any drastic effect on the water quality profile due to non-polluted characteristics of its sources.

1983-03-01

422

Polyphenol rich botanicals used as food supplements interfere with EphA2-ephrinA1 system

Skin sensitization is the most commonly reported occupational illness, causing much suffering to a wide range of people. Identification and labeling of environmental allergens is urgently required to protect people from skin sensitization. The guinea pig maximization test (GPMT) and murine local lymph node assay (LLNA) are the two most important in vivo models for identification of skin sensitizers. In order to reduce the number of animal tests, quantitative structure-activity relationships (QSARs) are strongly encouraged in the assessment of skin sensitization of chemicals. This paper has investigated the skin sensitization potential of 162 compounds with LLNA results and 92 compounds with GPMT results using a support vector machine. A particle swarm optimization algorithm was implemented for feature selection from a large number of molecular descriptors calculated by Dragon. For the LLNA data set, the classification accuracies are 95.37% and 88.89% for the ...

2009-07-17

423

British Library Electronic Table of Contents (United Kingdom)

The Eph tyrosine kinase receptors and their ephrin ligands play a central role in several human cancers and their deregulated expression or function promotes tumorigenesis, inducing aggressive tumor phenotypes. Green tea extracts (GTE) have been recently found to inhibit Eph-kinase phosphorylation. In order to evaluate the potential contribution of edible and medicinal plants on EphA2-ephrinA1 modulation, 133 commercially available plant extracts used as food supplements, essential and fixed oils were screened with an ELISA-based binding assay. Nine plant extracts, rich of polyphenols, reversibly inhibited binding in a dose-dependent manner (IC"5"0 0.83-24@mg/ml). Functional studies on PC3 prostate adenocarcinoma cells revealed that active extracts antagonized ephrinA1-Fc-induced EphA2-pho...

2011-01-01

424

Photobiological activity of marmesin (5-B-hydroxyisopropyl-4-5 dihydrofurocoumarin) in Chinese hamster V79 cells

Phosphate/UO{sub 2} interaction during in vitro dissolution of inhalable particles in a medium simulating the phagolysosomal compartment

Marmesin was isolated from the medicinal plant, Afraegle paniculata. Its cytotoxicity and mutagenicity in Chinese hamster V79 cells when sensitized to near ultraviolet (NUV) and long wavelength ultraviolet light or black light (BL) were assayed. Marmesin was extremely cytotoxic in the dark. This cytotoxicity was photoenhanced in NUV and BL; the photoenhanced lethality being higher in NUV than in BL. The LD/sub 50/ of marmesin under NUV and BL photosensitization were 0.002 ..mu..M and (0.012 ..mu..M), respectively. In the absence of NUV and BL, marmesin's LD/sub 50/ was 0.013 ..mu..M.NUV and BL without marmesin were not significantly cytotoxic at the fluence rates of 0.29 W/m/sup 2/ and 4.2 W/m/sup 2/, respectively, for up to 20 min. In contrast to the observed high cytotoxicity of marmesin, its mutagenicity at the HGPRT locus (Asub(z)Gsup(r)) was weak. The implication of this result in the high incidence of skin cancer in Nigeria in which A. paniculata is ...

1983-10-01

425

Phenolics from Glycyrrhiza glabra roots and their PPAR-g ligand-binding activity

Qualitative and quantitative methods have been developed to relate phosphate/UO{sub 2} interactions and U dissolution, in vitro, in a medium containing 0 or 10 mM phosphate concentrations at pH 5. This was performed by Energy Dispersive X ray Spectrometry (EDS) at 200 kV on entire particles and by fluorimetric measurement. The U dissolution rate was larger in a medium without phosphate than in a medium containing 10 mM phosphate. During the first day, the initial U dissolution involved 80.55% and 91.13% of the powder in media with and without phosphate respectively. From 5 to 15 d after beginning the assay, the dissolution half-times were longer than 92 d and 22 d for media with and without phosphate. EDS analyses have shown a constant P/U X ray intensity ratio for reference U{sub 2}O(PO{sub 4}){sub 2} for the same range of particle diameter. A gradual increase of this ratio was measured for UO{sub 2} incubated in a 10 mM phosphate medium throughout the experiment. ...

1998-07-01

426

British Library Electronic Table of Contents (United Kingdom)

Bioassay-guided fractionation of the EtOH extract of licorice (Glycyrrhiza glabra roots), using a GAL-4-PPAR-g chimera assay method, resulted in the isolation of 39 phenolics, including 10 new compounds (1-10). The structures of the new compounds were determined by analysis of their spectroscopic data. Among the isolated compounds, 5prime-formylglabridin (5), (2R,3R)-3,4prime,7-trihydroxy-3prime-prenylflavane (7), echinatin, (3R)-2prime,3prime,7-trihydroxy-4prime-methoxyisoflavan, kanzonol X, kanzonol W, shinpterocarpin, licoflavanone A, glabrol, shinflavanone, gancaonin L, and glabrone all exhibited significant PPAR-g ligand-binding activity. The activity of these compounds at a sample concentration of 10mg/mL was three times more potent than that of 0.5mM troglitazone.

2010-01-01

427

Pharmacokinetics of buspirone as determined by ex vivo (/sup 3/H)-DPAT binding

Organization and management activities in the nuclear power industry

Ex vivo (/sup 3/H)-8-hydroxy-2-(di-n-propylamino)-tetraline ((/sup 3/H)-DPAT) binding to the hippocampus has been utilized to determine the pharmacokinetic parameters of buspirone after i.v. and oral administration of this drug to rats. Intravenous buspirone rapidly penetrated the brain as demonstrated by a maximum inhibition of (/sup 3/H)-DPAT binding at 1 min. Elimination of drug from the brain was biphasic, with a first component half-life of 24.8 min and a second component half-life of 96 min. Oral buspirone at 3 times the i.v. dose produced less than one-third the maximum inhibition of (/sup 3/H)-DPAT binding compared to that observed with i.v. buspirone. The pharmacokinetic parameters of buspirone observed in the present study are in agreement with those reported previously. Thus, the ex vivo binding assay could be utilized to determine the bioavailability of the drug to the brain, and its duration of action. 20 references, 2 figures, 5 tables.

1988-01-01

428

Oil shale resonances of the Green River formation in the northeastern part of the Green River Basin, Wyoming

The purpose of organization and management development activities in the commercial nuclear power industry is to foster high levels of power plant performance and safety through improved human performance. The NRC has been working to develop assessment tools to assay the effects of organizational factors on plant safety. The utility industry has been working on initiatives targeting individual accountability, the improvement of plant performance and the elimination of the items identified through the NRC assessment process. Organization and management activities do not focus on industry organizational charts, but on the personnel processes and dimensions (factors) that affect safety and economic performance. As individual terms these activities are often combined and referred to as organizational factors. As an area of study, organizational factors has become more prominent as the industry emphasis has switched in recent years from hardware issues related to safety ...

1994-04-01

429

Novel high-throughput screening system for identifying STAT3-SH2 antagonists

Oil shale resource data are provided for a 1300-square mile area around Farson, Wyoming. The data were derived from analyses of samples from 14 core holes drilled by the US Bureau of Mines and the Department of Energy, plus 4 core holes drilled by private companies. Fischer assay oil yield data for nine stratigraphic resource units are tabulated, correlated, and contoured to provide an assessment of shale oil potential in the area. Oil shale resources in the northeastern part of the Green River Basin occur in the Laney Member and the upper part of the Wilkins Peak Member of the Eocene Green River Formation. A total in-place shale oil resource of 137 billion barrels was calculated for the area. Four stratigraphic resource units have average oil yields of 12 to 20+ gallons per ton within the area of investigation, and in some parts a section up to 275 feet thick averages as much as 15.9 gallons per ton. The thickness of overburden on the richest resource unit ranges ...

1988-03-01

430

New Lactone and Xanthone Derivatives Produced by a Mangrove Endophytic Fungus Phoma sp. SK3RW1M from the South China Sea

Constitutive activation of the oncogenic transcription factor STAT3 frequently occurs in various human malignancies. STAT3 activation involves dimerization via intermolecular pTyr-SH2 interaction. Thus, antagonizing this interaction is a feasible approach to inhibit STAT3 activation for cancer therapy. In order to identify selective STAT3 inhibitors, we developed a biochemical HTS system based on AlphaScreen technology, which measures the abilities of test compounds to antagonize pTyr-SH2 interactions. We screened our chemical libraries using this system and identified 5,15-diphenylporphyrin (5,15-DPP) as a selective STAT3-SH2 antagonist. Selective inhibition of STAT3 nuclear translocation and DNA biding activity was observed in cells treated with 5,15-DPP. IL-6-dependent dimerization of STAT3, c-myc promoter binding and c-myc protein expression were all suppressed by 5,15-DPP, whereas no decrement in either expression or phosphorylation level of STAT3 was observed. Thus, the HTS ...

2009-03-13

431

British Library Electronic Table of Contents (United Kingdom)

A new lactone, 1,8-dihydroxy-10-methoxy-3-methyldibenzo[b,e]oxepine-6,11-dione (1), and two new xanthones, 1-hydroxy-8-(hydroxymethyl)-6-methoxy-3-methyl-9H-xanthen-9-one (2) and 1-hydroxy-8-(hydroxymethyl)-3-methoxy-6-methyl-9H-xanthen-9-one (3), were isolated from a mangrove endophytic fungus Phoma sp. SK3RW1M collected from the South China Sea. This is the first report on xanthone derivatives isolated as secondary metabolites from Phoma species. Their structures were elucidated by spectroscopic methods, mainly 1D- and 2D-NMR techniques, and the structure of compound 2 was confirmed by X-ray crystallography. Cytotoxicity assays showed that compounds 1-3 were inactive against KB and KBv200 cells.

2010-01-01

432

Negative regulation of {beta}-catenin/Tcf signaling by naringenin in AGS gastric cancer cell

Natural killer cell activity influences outcome after T cell depleted stem cell transplantation from matched unrelated and haploidentical donors

Functional activation of {beta}-catenin/Tcf signaling plays an important role in early events in carcinogenesis. We examined the effect of naringenin against {beta}-catenin/Tcf signaling in gastric cancer cells. Reporter gene assay showed that naringenin inhibited {beta}-catenin/Tcf signaling efficiently. In addition, the inhibition of {beta}-catenin/Tcf signaling by naringenin in HEK293 cells transiently transfected with constitutively mutant {beta}-catenin gene, whose product is not phosphorylated by GSK3{beta}, indicates that its inhibitory mechanism was related to {beta}-catenin itself or downstream components. To investigate the precise inhibitory mechanism, we performed immunofluorescence, Western blot, and EMSA. As a result, our data revealed that the {beta}-catenin distribution and the levels of nuclear {beta}-catenin and Tcf-4 proteins were unchanged after naringenin treatment. Moreover, the binding activities of Tcf complexes to consensus DNA were not ...

2005-09-30

433

British Library Electronic Table of Contents (United Kingdom)

Lytic activity and recovery of natural killer (NK) cells was monitored in pediatric patients with leukemias (ALL, AML, CML, JMML) and myelodysplastic syndromes after transplantation of T cell depleted stem cells from matched unrelated (n = 18) and mismatched related (haploidentical, n = 29) donors. CD34 + selection with magnetic microbeads resulted in 8 x 10^3/kg residual T cells. No post-transplant immune suppression was given. NK cells recovered rapidly after transplantation (300 CD56+/@mL at day 30, median), whereas T cell recovery was delayed (median: 12 CD3+/@mL at day 90). NK activity was measured as specific lysis of K 562 targets several times (mean: 3 assays per patient). Four temporal patterns of lytic activity could be differentiated: consistently low, consistently high, decreas...

2011-01-01

434

Modulating factors in the expression of radiation-induced oncogenic transformation

Modulating factors in the expression of radiation-induced oncogenic transformation

Many assays for oncogenic transformation have been developed ranging from those in established rodent cell lines where morphological alteration is scored, to those in human cells growing in nude mice where tumor invasiveness is scored. In general, systems that are most quantitaive are also the least relevant in terms of human carcinogenesis and human risk estimation. The development of cell culture systems has made it possible to assess at the cellular level the oncogenic potential of a variety of chemical, physical and viral agents. Cell culture systems afford the opportunity to identify factors and conditions that may prevent or enhance cellular transformation by radiation and chemicals. Permissive and protective factors in radiation-induced transformation include thyroid hormone and the tumor promoter TPA that increase the transformation incidence for a given dose of radiation, and retinoids, selenium, vitamin E, and 5-aminobenzamide that inhibit the expression ...

1990-08-01

435

Ligands for the peroxisome proliferator-activated receptor-#gamma# have inhibitory effects on growth of human neuroblastoma cells in vitro

Many assays for oncogenic transformation have been developed ranging from those in established rodent cell lines where morphological alteration is scored, to those in human cells growing in nude mice where tumor invasiveness is scored. In general, systems that are most quantitaive are also the least relevant in terms of human carcinogenesis and human risk estimation. The development of cell culture systems has made it possible to assess at the cellular level the oncogenic potential of a variety of chemical, physical and viral agents. Cell culture systems afford the opportunity to identify factors and conditions that may prevent or enhance cellular transformation by radiation and chemicals. Permissive and protective factors in radiation-induced transformation include thyroid hormone and the tumor promoter TPA that increase the transformation incidence for a given dose of radiation, and retinoids, selenium, vitamin E, and 5-aminobenzamide that inhibit the expression ...

1990-01-01

436

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

The thiazolidinedione (TZD) or glitazone class of peroxisome proliferator-activated-#gamma# (PPAR-#gamma#) ligands not only induce adipocyte differentiation and increase insulin sensitivity, but also exert growth inhibitory effects on several carcinoma cell lines in vitro as well as in vivo. In the current study the in vitro effect of four PPAR-#gamma# agonists (ciglitazone, pioglitazone, troglitazone, rosiglitazone) on the cell growth of seven human neuroblastoma cell lines (Kelly, LAN-1, LAN-5, LS, IMR-32, SK-N-SH, SH-SY5Y) was investigated. Growth rates were assessed by a colorimetric XTT-based assay kit. Expression of PPAR-#gamma# protein was examined by immunohistochemistry and Western blot analysis. All glitazones inhibited in vitro growth and viability of the human neuroblastoma cell lines in a dose-dependent manner showing considerable effects only at high concentrations (10 #mu#M and 100 #mu#M). Effectiveness of the glitazones on neuroblastoma cell growth ...

2005-09-15

437

Inventory determination of low and intermediate level radioactive waste of Paks Nuclear Power Plant origin

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

2010-04-15

438

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

In the execution of disposal of low and intermediate level radioactive wastes, it is important to evaluate accurately the kind and quantity of each radionuclide in the wastes. For such an evaluation, correlation of non-gamma-emitting nuclides based on gamma-emitting nuclides is recommended and regarded as a partial method. This method necessitates a completion of a highly accurate and reliable nondestructive assay system of gamma-emitting nuclides for partical use. In 1992, in support of the new waste disposal program in Hungary, Paks NPP initiated a waste characterization program to determine the radiological properties of its rad wastes. A segmented gamma scanning system has been set up to measure the gamma-emitting nuclides in 2000 litre low level drums following in-drum compaction. In the framework of the program a radiochemical analysis sub-program was stated to determine the long-lived non-gamma emitting radionuclides, mainly those listed in US regulatory ...

439

Impact of the Sea Empress oil spill on lysosomal stability in mussel blood cells

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

2005-07-01

440

Identification of tumor-initiating cells in a p53-null mouse model of breast cancer.

Coastal zones are among the most productive and vulnerable areas on the planet. An example of impact on these fragile environments was shown in the case of the Sea Empress oil tanker, which ran aground in the Bristol Channel in 1996, spilling 72,000 tonnes of Forties crude oil. The objective was to investigate the sub-lethal cellular pathology and tissue hydrocarbon contamination in marine mussel populations, 4 months after the initial spill, using the neutral red retention (NRR) assay for lysosomal stability in blood cells. NRR was reduced in mussels, and indicative of cell injury, from the two sites closest to the spill in comparison with more distant and reference sites. Lysosomal stability was inversely correlated with polycyclic aromatic hydrocarbon concentrations in mussel tissues. Reduced lysosomal stability has previously been shown to contribute to impaired immunocompetence and to autophagic loss of body tissues. The use of this type of technique is ...

2000-07-01

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

441

Identification of single nucleotide polymorphisms in candidate genes for growth and reproduction in a nonmodel organism; the Atlantic cod, Gadus morhua

Using a syngeneic p53-null mouse mammary gland tumor model that closely mimics human breast cancer, we have identified, by limiting dilution transplantation and in vitro mammosphere assay, a Lin(-)CD29(H)CD24(H) subpopulation of tumor-initiating cells. Upon subsequent transplantation, this subpopulation generated heterogeneous tumors that displayed properties similar to the primary tumor. Analysis of biomarkers suggests the Lin(-)CD29(H)CD24(H) subpopulation may have arisen from a bipotent mammary progenitor. Differentially expressed genes in the Lin(-)CD29(H)CD24(H) mouse mammary gland tumor-initiating cell population include those involved in DNA damage response and repair, as well as genes involved in epigenetic regulation previously shown to be critical for stem cell self-renewal. These studies provide in vitro and in vivo data that support the cancer stem cell (CSC) hypothesis. Furthermore, this p53-null mouse mammary tumor model may allow us to identify new ...

2008-06-15

442

DEFF Research Database (Denmark)

Recent technological developments have facilitated intensified searches for genetic markers under selection in nonmodel species. Here, we present an approach for the identification of candidate gene variation in nonmodel organisms. We report on the characterization of 82 single nucleotide polymorphisms (SNPs) and on the development of a specific genotyping assay for 30 SNPs in 18 candidate genes for growth and reproduction in Atlantic cod (Gadus morhua). These markers can be used for scanning natural populations for signatures of selection in both contemporary and archived historical samples, for example in retrospective studies assessing the effects of environmental changes, such as increasing temperatures, and selection imposed by high fishing pressure. Furthermore, these gene markers may be of interest to aquaculture, serving as a starting point for linking phenotypic traits important for productivity with genotypes and potentially be of use for marker-assisted ...

2011-01-01

443

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

Human complement protein C99 is a calcium binding protein

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.

1982-12-15

444

Free release waste characterisation during the decommissioning of windscale Pile 2 Chimney

Human complement protein C9 is shown to be a metalloprotein that binds 1 mol of Ca"2"+/mol of C9 with a dissociation constant of 3 #mu#m as measured by equilibrium dialysis. Incubation with EDTA removes the bound calcium, resulting in a apoprotein with decreased thermal stability. This loss in stability leads to aggregation and, therefore, to loss of hemolytic activity upon heating to a few degrees above the physiological temperature. Heat-induced aggregation of apoC9 can be prevented by salts that stabilize proteins according to the Hofmeister series of lyotropic ions, suggesting that the ion in native C9 may ligand with more than one structural element of domain of the protein. Ligand blotting indicates that the calcium binding site is located in the amino-terminal half of the protein. Removal of calcium by inclusion of EDTA in assay mixtures has no effect on the hemolytic activity of C9, and its capacity to bind to C8 in solution, or to small unilamellar lipid ...

445

FMRFamide-like immunoreactivity in rat brain: development of a radioimmunoassay and its application in studies of distribution and chromatographic properties

The decommissioning of windscale Pile 2 Chimney resulted in the removal of one of most historically significant and prominent features of the Sellafield site. The project was the first large scale concrete ''free release'' operation to be undertaken on the Sellafield nuclear site, producing 4500 tons of concrete of which 3000 tons was demonstrated to be ''free release''. The paper describes the radiometric techniques employed in the characterisation and segregation of the concrete into low level waste (LLW), very low level waste (VLLW) and free release categories. It examines the robust solutions that were developed to meet the technical and regulatory challenges of the project, which included the definition of free release, the selection of averaging volumes, the testing and validation of the monitoring systems employed for ton quantities of concrete and the identification and removal of small numbers of fuel particles from the free release waste stream. As a result of interest shown ...

2002-09-22

446

Expression profiles of precursor and mature microRNAs under dehydration and high salinity shock in Populus euphratica

A radioimmunoassay is described for the molluscan neuropeptide, Phe-Met-Arg-Phe-NH"2(FMRFamide). The antibody used is C-terminal-specific and shows slight but significant (1-2%) cross-reactivity with chicken pancreatic polypeptide (APP). The assay has been used to identify in rat brain extracts a pair of molecules that may represent mammalian counterparts of FMRFamide. Their concentrations were highest in spinal cord and hypothalamus (>10 pmol.g"-_1) and lowest in cerebellum and striatum (<3.5 pmol.g"-_1). The two immunoreactive peptides were separated on CM ion-exchange chromatography where they appeared to be less basic than FMRFamide. On Sephadex G50 gel filtration one eluted in a similar position to FMRFamide and the other slightly earlier suggesting it may be of higher molecular weight. The rat immunoreactive components do not correspond to previously described neuropeptides or hormones, and may be members of a new group of mammalian neuropeptides with ...

447

British Library Electronic Table of Contents (United Kingdom)

MicroRNAs (miRNAs) are small non-coding RNAs that play vital roles in plant abiotic stress responses via cleavage or translational inhibition of their target mRNAs. Populus euphratica is a typical stress-resistant sessile organism that grows in desert areas. Here, we identified sequences of 12 miRNA precursors from 11 families and 13 mature miRNAs from 12 families by PCR amplification in P. euphratica. To detect expression differences in mature miRNAs and their precursors under dehydration and high salinity shock in P. euphratica, we examined 14 miRNA precursors from 13 miRNA families and 17 mature miRNAs from 17 miRNA families using the SYBR Green RT?PCR assay. This is the first report of expression profiles for both precursor and mature miRNAs in P. euphratica. By profiling both the matu...

2011-01-01

448

Endogenous biosynthesis of arachidonic acid epoxides in humans: Increased formation in pregnancy-induced hypertension

Elimination of a new ampelovirus (GLRaV-Pr) and Grapevine rupestris stem pitting associated virus (GRSPaV) from two Vitis vinifera cultivars combining in vitro thermotherapy with shoot tip culture

Arachidonic acid is metabolized by means of P450 isoenzyme(s) to form epoxyeicosatrienoic acids (EETs) and their corresponding dihydroxy derivatives (DHETs). In the present study, we established the presence in human urine of 8,9-, 11,12-, and 14,15-EETs and their corresponding DHETs by developing quantitative assays and using negative ion, chemical ionization GC/MS and octadeuterated internal standards. Urinary excretion of 8,9- and 11,12-DHET increased in healthy pregnant women compared with nonpregnant female volunteers. By contrast, excretion of 11,12-DHET and 14,15-DHET, but not the 8,9-DHET regioisomer, increased even further in patients with pregnancy-induced hypertension. Intravenous administration of (3H)14,15-EET to three dogs markedly increased its DHET in plasma. The terminal half-life ranged from 7.9-12.3 min and the volume of distribution (3.5-5.3 liters) suggested limited distribution outside the plasma compartment. Negligible radioactivity was ...

1990-08-01

449

British Library Electronic Table of Contents (United Kingdom)

A new virus species designated as Grapevine leafroll associated virus-Pr (GLRaV-Pr), which is classified in a distinct phylogenetic group of the genus Ampelovirus (Closteroviridae), was recently characterized from Greek grapevine cultivars. Elimination studies of GLRaV-Pr were carried out in two grapevine cultivars, 'Mantilaria' and 'Prevezaniko', co-infected with Grapevine rupestris stem pitting associated virus (GRSPaV, Flexiviridae). Both viruses were detected by nested RT-PCR assays. Virus elimination was achieved by combining in vitro thermotherapy with meristem ( 0.2mm) or shoot tip culture ( 0.5cm). The survival and regeneration rate of meristems was very low. On the other hand, high survival rates were observed in the cultured shoot tips accompanied with high elimination rates for ...

2009-01-01

450

Electrochemical detection of DNA hybridization based on silver-enhanced gold nanoparticle label

Effect of the steam explosion pretreatment on enzymatic hydrolysis of eucalyptus wood and sweet sorghum baggages; Efecto del pretratamiento con explosion por vapor en la hidrolisis enzimatica de madera de eucalipto y bagazo de sorgo

An electrochemical detection method for analyzing sequence-specific DNA using gold nanoparticle DNA probes and subsequent signal amplification step by silver enhancement is described. The assay relies on the electrostatic adsorption of target oligonucleotides onto the sensing surface of the glassy carbon electrode (GCE) and its hybridization to the gold nanoparticle-labeled oligonucleotides DNA probe. After silver deposition onto gold nanoparticles, binding events between probe and target were monitored by the differential pulse voltammetry (DPV) signal of the large number of silver atoms anchored on the hybrids at the electrode surface. The signal intensity difference permits to distinguish between the match of two perfectly matched DNA strands and the near-perfect match where just one base pair was wrong. Coupled with this 'nanoparticle-promoted' reduction of silver signal amplification method, the sensitivity of this electrochemical DNA ...

2002-10-03

451

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

The effect of steam explosion treatment on the enzymatic hydrolysis yield of two different lignocellulosic substrates is studied. Raw materials have been pretreated in a pilot plant designed to work in batch and equipped with a reactor vessel of 2 1 working volume where biomass was heated at the desired temperature and then exploded and recovered in a cyclone. Temperatures from 190 to 230 degree celsius and reaction times from 2 to 8 min. have been assayed. The efficiency of the steam explosion treatment has been evaluated on the composition of the lignocellulosic materials as well as on their enzymatic hydrolysis yield using a cellulolytic complex from T. reesel. Results show a high solubilization rate of hemicelluloses and variable losses of cellulose and lignin depending on the conditions tested. Enzymatic hydrolysis yields of both substrates experimented remarkable increments, corresponding the highest values obtained to 210 degree celsius; 2 min. and 21O ...

1991-07-01

452

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

453

Effect of chronic fractionated low-dose gamma irradiation on division potential of human embryonic cells in vitro

Development of radiological emergency preparedness and biological dosimetry technology

We investigated the in vitro phenotypic transformation of human embryo (HE) cells that were repeatedly irradiated (7.5 cGy once a week) throughout their life-span. Irradiation was repeated until the cells had accumulated 195 cGy (equivalent to the 26th passage). Samples of cells were assayed for survival by colony formation, as well as for mutation at the hypoxanthine guanine phosphoribosyl transferase (HGPRT) locus and for transformation by focus formation. The life-span (mean number of population doublings) of multiply irradiated cells with a total dose of 97.5 cGy was slightly but significantly prolonged over that of controls. After HE cells had accumulated 195 cGy, the maximum number of divisions increased to 130-160% of the number in non-irradiated control cells. Transformed foci were not observed until cells had accumulated 97.5 cGy, and then increased with the increasing accumulation of radiation. However, no cells showed immortality or expressed a malignant ...

1991-12-01

454

Development of a multiplex PCR method for simultaneous detection of diagnostic DNA markers of five disease and pest resistance genes in potato

Large-scale field tracer experiments have been conducted on Ulchin and Wolsung nuclear sites for the purpose of validating FADAS and of analyzing the environmental characteristics around the nuclear site. The most influential factor in atmospheric dispersion is the meteorological condition. During the experiment, meteorological data were measured on the release point and the selected positions among sampling points. Once radioactive materials are released to the atmosphere, members of public may be exposed through the environmental media such as air, soil and foods. Therefore, to protect the public, adequate countermeasures should be taken at due time for those exposure pathways. Both processes of justification and optimization are applied to a countermeasure simultaneously for decision-making. The work scope of biological research for the radiation protection had contained the search of biological microanalytic methods for the assessment of health effect by radiation and toxic agents, ...

1999-04-01

455

British Library Electronic Table of Contents (United Kingdom)

Multiplex PCR is practically a reasonable choice for molecular marker-assisted selection in potato breeding. We had developed and were using a multiplex PCR method for selection of resistance genes to cyst nematode (H1), Potato virus X (Rx1) and late blight (R1 and R2). Since then, more reliable and tightly linked markers for H1 and R2, and a new marker for resistance to Potato virus Y (Ry chc ) were developed. In this article, all these superior markers, including a positive marker to eliminate PCR-failed samples, were incorporated into one multiplex PCR assay. Using the newly developed multiplex PCR technique, five plants potentially harboring all five resistance genes were selected from 96 hybrid plants approximately 5?h after DNA extraction, which is a third of the operation time compa...

2011-01-01

456

Determination of binary mixtures of analgesic and spasmolytic drugs in pure and dosage forms by derivative spectrophotometry.

Determination of baclofen enantiomers in pharmaceutical formulations using maltodextrin-based enantioselective, potentiometric membrane electrodes.

Binary mixtures of dipyrone and pitophenone hydrochloride are assayed by zero-crossing second- and third-derivative spectrophotometry and by ratio-spectra first- and second-derivative spectrophotometry. In the first method, calibration plots are linear at 266.5 and 302.5 nm (dipyrone, second derivative), and 257 and 286 nm (pitophenone second derivative) and 242 and 278.3 nm (dipyrone third derivative), and 228.5 and 300 nm (pitophenone, third-derivative). By the second method, lines of regression are linear at 235 and 262 nm (dipyrone, first derivative), and 229.5 and 288.5 nm (pitophenone, first-derivative), and 249.7 and 268 nm (dipyrone, second derivative), and 280.5 and 300 nm (pitophenone, second-derivative). In all methods calibration curves follow the Beer's law up to 40 microg/ml of each drug. LOD and LOQ values were calculated. The developed derivative spectrophotometric methods were applied to laboratory mixtures and to vials for these drugs. The ...

2003-10-15

457

Detecting exposure to environmental organic toxins in individual cells: towards development of a micro-fabricated device

Two enantioselective, potentiometric membrane electrodes (EPMEs) based on maltodextrins with different value of dextrose equivalent (DE) (maltodextrin I: DE 4.0-7.0; maltodextrin II: DE 16.5-19.5) were proposed for the assay of baclofen enantiomers in baclofen raw materials and from its pharmaceutical formulation, Norton-Baclofen tablets. The slopes of the electrode function of the proposed electrodes were 55.0 mV/pS-baclofen for maltodextrin I-based electrode and 59.0 mV/pR-baclofen for maltodextrin II-based electrode and the detection limits were 1.34 x 10(-6) mol l(-1) (S-baclofen) and 2.52 x 10(-10) mol l(-1) (R-baclofen), respectively. The surfaces of the electrodes are stable and easily renewable by simply polishing on alumina paper. PMID:15598435

2004-12-01

458

DYMAC communications system

A new method is being developed to quickly screen for the human exposure potential to polycyclic aromatic hydrocarbons (PAHs) and organochlorines (OCs). The development involves two key elements: identifying suitable signals that represent intracellular changes that are specific to PAH and OC exposure, and constructing a device to guide the biological cell growth so that signals from individual cells are consistent and reproducible. We are completing the identification of suitable signals by using synchrotron radiation-based (SR) Fourier-transform infrared (FTIR) spectromicroscopy in the mid-infrared region (4000-400 cm-1). Distinct changes have been observed in the IR spectra after treatment of human cells in culture medium with PAHs and OCs. The potential use of this method for detecting exposure to PAHs and OCs has been tested and compared to a reverse transcription polymerase chain reaction (RT-PCR) assay that quantifies increased expression of the CYP1A1 gene ...

1999-01-10

459

DETERMINATION OF 237NP AND PU ISOTOPES IN LARGE SOIL SAMPLES BY INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY

The DYMAC Communications System is part of a nuclear safeguards system called DYMAC - short for DYnamic Materials ACcountability - that gathers accountability information at the Los Alamos Scientific Laboratory (LASL) Plutonium Processing Facility. The communications system handles transmissions between data-entry terminals and nondestructive assay (NDA) instruments located in the facility, and a computer located in an adjacent building. System design emphasizes reliability rather than high speed to ensure the integrity of data transmissions. This manual is directed toward the person responsible for maintaining the DYMAC Communication System. It describes the components that make up the communications network, explains how they operate, and gives detailed information about all of the connections. Many of the system components are commercially available; some have been modified at LASL for DYMAC purposes; others were designed and fabricated at LASL. This manual ...

1980-01-01

460

Cytotoxic Naphtho--pyrones from the Mangrove Endophytic Fungus Aspergillus tubingensis (GX1-5E)

A new method for the determination of {sup 237}Np and Pu isotopes in large soil samples has been developed that provides enhanced uranium removal to facilitate assay by inductively coupled plasma mass spectrometry (ICP-MS). This method allows rapid preconcentration and separation of plutonium and neptunium in large soil samples for the measurement of {sup 237}Np and Pu isotopes by ICP-MS. {sup 238}U can interfere with {sup 239}Pu measurement by ICP-MS as {sup 238}UH{sup +} mass overlap and {sup 237}Np via {sup 238}U peak tailing. The method provides enhanced removal of uranium by separating Pu and Np initially on TEVA Resin, then transferring Pu to DGA resin for additional purification. The decontamination factor for removal of uranium from plutonium for this method is greater than 1 x 10{sup 6}. Alpha spectrometry can also be applied so that the shorter-lived {sup 238}Pu isotope can be measured successfully. {sup 239}Pu, {sup 242}Pu and {sup 237}Np were measured ...

2010-07-26

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

461

British Library Electronic Table of Contents (United Kingdom)

Abstract Four new dimeric naphtho--pyrones, named rubasperone D (1), rubasperone E (2), rubasperone F (3), and its atropisomer rubasperone G (4), together with four known monomeric naphtho--pyrones, TMC 256 A1 (5), rubrofusarin B (6), fonsecin (7), and flavasperone (8), were isolated from the mangrove endophytic fungus Aspergillus tubingensis (GX1-5E) cultivated in solid rice medium. Their structures were elucidated by spectroscopic methods, including IR, 1D- and 2D-NMR, and MS. In the in vitro cytotoxicity assays, 5 displayed inhibitory activities against tumor cell lines of MCF-7, MDA-MB-435, Hep3B, Huh7, SNB19, and U87-MG with IC50 values between 19.92 and 47.98-M. Compounds 1, 6, and 8 also showed mild cytotoxic activity.

2011-01-01

462

Cyclin-Dependent Kinase Inhibitor p16 and p21 Expression, and Cell Cycle Change in Human Lens Epithelial Cell Line SRA 01/04 following Contact Inhibition in Normal Culture

British Library Electronic Table of Contents (United Kingdom)

Abstract Purpose: To describe the pattern of expression of the cyclin-dependent kinase inhibitors (CDKIs) p16, p21 and p27, and the cell cycle in SRA 01/04 cells relative to contact inhibition. Methods: SRA 01/04 cells were grown to overconfluence under normal conditions. At various phases of the cell growth, cells were assayed by flow cytometry and Western blotting for the expression of CDKIs. Results: Expression of p16 was detected from early logarithmic growth to stationary phases, during which the number of cells in G0/G1 increased from 46 to 69%. Expression of p21 was detected only during the overgrowth phase, when 60% of the cells were in G0/G1. Expression of p27 was not observed in SRA 01/04 cells. Conclusions: p16 expression was likely mediated by G0/G1 arrest to induce contact inh...

2011-01-01

463

Clinical evaluation of serum ferritin to iron ratio in malignant diseases

Characterization of the xanthine-binding site on R/sub a/ and R/sub i/ subtypes of the adenosine receptor

Serum ferritin levels were measured in 72 normal subjects and in 214 cases with various diseases by an immunoradiometric assay. In normal subjects, the serum ferritin levels were 27-230 ng/ml. Elevated serum ferritins were observed in most cases with iron excess and acute hepatitis. Markedly elevated levels were found in the majority of cases with acute leukemia, malignant lymphoma, hepatoma, and pancreatic cancer. High ferritin levels were also found in other malignant diseases. However, the range overlapped broadly with that of nonmalignant diseases. The serum ferritin correlated significantly with serum iron in normals and in those with iron deficiency anemia. In most nonmalignant cases, the serum ferritin and iron levels distributed on a regression line obtained from levels in normals and those with iron deficiency anemia. However, 92% of the malignant cases showed a serum ferritin to iron ratio higher than that of normal subjects. The estimation of the serum ...

1981-07-01

464

Changes of hypoxia inducible factor-1 ? in hepatoma cells irradiated by gamma ray and their mechanism

Methylxanthines and their derivatives are antagonists at cell surface adenosine receptors. They report here a systematic study of xanthine structure-activity relationships which compares potency at two adenosine receptor subtypes, R/sub a/ and R/sub i/. Adenylate cyclase stimulation (R/sub a/ in platelet membranes) and inhibition (R/sub i/ in adipocyte membranes) were used as models of receptor activation. K/sub i/ values were obtained by Schild analysis. The orders of potency of the xanthines to attenuate the effects of adenosine analogues were similar to those previously reported. Earlier work utilizing radioligand binding (R/sub i/ and (/sup 3/H) cAMP formation (R/sub a/) claimed that IIX and PACPX are at least 10 and 400 fold, respectively, more potent at R/sub i/ than at R/sub a/. However, in their assays which utilize modulation of receptor-coupled adenylate cyclase, the xanthines show little specificity for either adenosine receptor subtype.

1986-05-01

465

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

This paper reports regulation of hypoxia inducible factor-1 ? (HIF-1 ?)in hepatoma cells by irradiation. Cobalt chloride (CoCl2), a chemical mimic agent for hypoxia research, was utilized to induce the stable expression of HIF-1 ? in HepG2 cells. The HepG2 cells were irradiated to different doses to observe the changes of HIF-1 ?. The level of intracellular reactive oxygen species (ROS) was assayed by fluorescent microscope and flow cytometry (FCM). The results showed that there were obvious changes in expression of HIF-1 ? after HepG2 cells exposed to radiation, and the changes were positively related with the irradiation dose from 1 Gy to 5 Gy. Moreover, contents of incellular ROS were negatively correlated with above levels of HIF-1 ? from 1 Gy to 3 Gy. The results indicate that irradiation may enhance hypoxic cells HIF-1 ?, and the reduction of intracellular ROS can contribute to the regulation of ionizing radiation on HIF-1 ?. (authors)

2007-10-01

466

British Library Electronic Table of Contents (United Kingdom)

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

2011-01-01

467

Analysis of S-adenosylmethionine and related sulfur metabolites in bacterial isolates of Pseudomonas aeruginosa (BAA-47) by liquid chromatography/electrospray ionization coupled to a hybrid linear quadrupole ion trap and Fourier transform ion cyclotron resonance mass spectrometry

British Library Electronic Table of Contents (United Kingdom)

A comprehensive and highly selective method for detecting in bacterial supernatants a modified sulfur nucleoside, S-adenosyl-L-methionine (SAM), and its metabolites, i.e., S-adenosylhomocysteine (SAH), adenosine (Ado), 5prime-deoxy-5prime-methylthioadenosine (MTA), adenine (Ade), S-adenosyl-methioninamine (dcSAM), homocysteine (Hcy) and methionine (Met), was developed. The method is based on reversed-phase liquid chromatography with positive electrospray ionization (ESI+) coupled to a hybrid linear quadrupole ion trap (LTQ) and 7-T Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS). A gradient elution was employed with a binary solvent of 0.05 M ammonium formate at pH 4 and acetonitrile. The assay involves a simultaneous cleanup of cell-free bacterial broths by solid-pha...

2009-01-01

468

Activation of fat cell adenylate cyclase by protein kinase C

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

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 ...

1986-05-01

469

Radiation 2006. In association with the Polymer Division, Royal Australian Chemical Institute. Incorporating the 21st AINSE Radiation Chemistry Conference and the 18th Radiation Biology Conference, conference handbook

The 2- and 8-azido trimer 5'-triphosphate photoprobes of 2-5A have been enzymatically synthesized from [#gamma#-"3"2P]2-azidoATP and [#alpha#-"3"2P]8-azidoAPT by 2-5A synthetase from rabbit reticulocyte lysates. Identification and structural determination of the 2- and 8-azido adenylate trimer 5'-triphosphates were accomplished by enzymatic hydrolyses with T2 RNase, snake venom phosphodiesterase, and bacterial alkaline phosphatase. Hydrolysis products were identified by HPLC and PEI-cellulose TLC analyses. The 8-azido photoprobe of 2-5A displaces p_3A_4["3"2P]pCp from RNase L with affinity equivalent to p_3A_3. The 8-azido photoprobe also activates RNase L to hydrolyze poly(U)["3"2P]pCp 50% at 7 /times/ 10/sup /minus/9/ M in core-cellulose assays. The 2- and 8-azido photoprobes and authentic p_3A_3 activate RNase L to cleave 28S and 18S rRNA to specific cleavage products at 10/sup /minus/9/ M in rRNA cleavage assays. The nucleotide binding ...

470

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

Full text: An adaptive response is a decreased biological effect induced by a priming radiation dose given prior to a challenge dose. Adaptive responses contradict the linear-nothreshold model of risk estimation. The pKZ1 mouse chromosomal inversion assay is an extremely sensitive assay for studying the mutagenic effect of low dose radiation. A non-linear dose response for chromosomal inversion has been observed in pKZ1 spleen and prostate after a single whole body irradiation with doses between 1?Gy and 10mGy. Doses between 5-10?Gy resulted in an induction in inversions and doses between 1-10mGy resulted in a reduction below endogenous inversion frequency. These results suggest that doses in the 1-10 mGy range cause host responses which overcompensate by not only preventing inversions that would normally occur as a result of the low doses of radiation but also by preventing some of the endogenous inversions that would have occurred in the ...

2006-04-01

471

An ELISA-based high throughput protein truncation test for inherited breast cancer

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

2006-07-01

472

Use of sting-response techniques for simulate diagnostics in human esophagus; Uso de tecnicas estimulo-respuesta para simular diagnosticos en esofago humano

IntroductionBreast cancer is the most diagnosed and second leading cause of cancer deaths in the U.S. female population. An estimated 5 to 10 percent of all breast cancers are inherited, caused by mutations in the breast cancer susceptibility genes (BRCA1/2). As many as 90% of all mutations are nonsense mutations, causing a truncated polypeptide product. A popular and low cost method of mutation detection has been the protein truncation test (PTT), where target regions of BRCA1/2 are PCR amplified, transcribed/translated in a cell-free protein synthesis system and analyzed for truncated polypeptides by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography. We previously reported a novel High Throughput Solid-Phase PTT (HTS-PTT) based on an enzyme-linked immunosorbent assay (ELISA) format that eliminates the need for radioactivity, SDS-PAGE and subjective interpretation of the results. Here, we report the next generation HTS-PTT ...

2010-10-04

473

Toxicity of N-substituted aromatics to acetoclastic methanogenic activity in granular sludge

In this work a study of simulation of the gamma graphic studies that are carried out in human esophagus in the Dept. of Nuclear Medicine of the 'Celestino Hernandez Robau Hospital of Santa Clara is presented. For the investigation tubular reactors were used and sting-response techniques with radioactive tracer of Technetium 99 metastable to a concentration of 1 mCi and several flows were applied. The distribution curves of residences times were obtained, those that respond to an equation of the type: Y = A + B exp (- exp((x-C)/D)) - ((x-C/D)+1). They were also carried out, optimizations studies of the doses of the radioactive to give to the patients from 1 mCi (that is the one used in studies) up to 0,5 mCi, and the influences on the obtained distributions of residence time were analyzed. It was confirmed the possibility to lower the doses with clear information of the signal. It was also carried out a simulation of the attenuation of the radiations that takes place in the ...

2003-07-01

474

The relationship between seminal plasma zinc levels and high molecular weight zinc binding protein and sperm motility in Iraqi infertile men

N-substituted aromatics are important priority pollutants entering the environment primarily through anthropogenic activities associated associated with the industrial production of dyes, explosives, pestides, and pharmaceuticals. Anaerobic treatment of wastewaters discharged by these industries could potentially be problematical as a result of the high toxicity of N-substituted aromatics. The objective of this study was to examine the structure-toxicity relationship of N-substituted aromatic compounds to acetoclastic methanogenic bacteria. The toxicity was assayed to serum flasks by measuring methane production in granular sludge. Unacclimated cultures were used to minimize the biotransformation of the toxic organic chemicals during the test. The nature and the degree of the aromatic substitution were observed to have a profound effect on the toxicity of the test compound. Nitroaromatic compounds were, on the average, over 500-fold more toxic than their ...

1995-11-01

475

The growth factor from plerocercoid larvae of the tapeworm, Spirometra mansonoides, stimulates growth but is not diabetogenic.

To evaluate the relationship between sperm motility and total seminal plasma zinc concentration and high molecular weight zinc bound protein values in infertile Iraqi men. A case-control study was conducted at the Chemistry and Biochemistry Department, College of Medicine, Al-Nahrain University, Baghdad, Iraq between March 2005 to February 2006. The subjects for the study included 60 infertile male patients who were recruited Al-Kadhimiya Teaching Hospital, and Institute of Embryo Research and Infertility Treatment, Baghdad, Iraq. They were categorized according to their seminal parameters to oligozoospermia (n=32), azoospermia (n=22), and asthenozoospermia (n=6). Thirty nine fertile men (age range 31.87 +/- 3.76 years) were selected as controls, whose partners had conceived within the last year before participation with this study, and having normal spermiogram parameters. Seminal plasma zinc concentration and high molecular weight zinc binding proteins (HMW-Zn) were ...

476

Structure-function studies on inhibitory activity of Bungarus multicinctus protease inhibitor-like protein on matrix metalloprotease-2, and invasion and migration of human neuroblastoma SK-N-SH cells.

A factor produced by plerocercoids of the tapeworm Spirometra mansonoides is similar to human growth hormone (hGH) in that it stimulates body growth, binds to hGH receptors, cross-reacts with anti-hGH antibodies, and has lactogenic and insulin-like activities. The purpose of this study was to determine whether plerocercoid growth factor (PGF) is similar to hGH in expressing diabetogenic activity in the genetically obese (ob/ob) mouse. To determine an effective dose for use in the obese mice, the ability of daily injections of PGF to stimulate growth of phenotypically normal mice of the same strain was assessed in a 10-day weight gain assay. Injections of PGF stimulated a dose-dependent weight gain (r = 0.83) and 25 ng eq/day of PGF stimulated a response not significantly different from that produced by 100 micrograms of bovine growth hormone/day. Diabetogenicity was assessed using fasting blood glucose and glucose tolerance tests in obese mice that had been ...

1989-06-01

477

Single protein omission reconstitution studies of tetracycline binding to the 30S subunit of Escherichia coli ribosomes

In view of the findings that several Kunitz-type protein inhibitors suppress tumor invasion and metastasis, the aim of the present study is to explore whether Bungarus multicinctus protease inhibitor-like protein-2 (PILP-2) and PILP-3 exhibit anti-tumor activity. Although approximately 28% of amino acid substitutions occurred between PILP-2 and PILP-3, molecular modeling suggested that PILP-2 and PILP-3 shared similar folded structures. Unlike PILP-2, PILP-3 showed a notable activity in abolishing migration and invasion of human neuroblastoma SK-N-SH cells. The ability of PILP-3 to inhibit matrix metalloprotease-2 (MMP-2) activity was higher than that of PILP-2. Pull-down assay revealed protein-protein interaction between PILP-3 and MMP-2. In contrast to mutation on N-terminal region, replacement of amino acids at C-terminus attenuated notably the ability of PILP-3 to inhibit cell invasion, cell migration and MMP-2 activity as well as the binding capability of ...

2009-08-23

478

Resistance to powdery mildew in Spanish barley landraces is controlled by different sets of quantitative trait loci.

In previous work the authors showed that on photolysis of Escherichia coli ribosomes in the presence of ({sup 3}H)tetracycline (TC) the major protein labeled is S7, and they presented strong evidence that such labeling takes place from a high-affinity site related to the inhibitory action of TC. In this work they use single protein omission reconstitution (SPORE) experiments to identify those proteins that are important for high-affinity TC binding to the 30S subunit, as measured by both cosedimentation and filter binding assays. With respect to both sedimentation coefficients and relative Phe-tRNA{sup Phe} binding, the properties of the SPORE particles they obtain parallel very closely those measured earlier, with the exception of the SPORE particle lacking S13. A total of five proteins, S3, S7, S8, S14, and S19, are shown to be important for TC binding, with the largest effects seen on omission of proteins S7 and S14. Determination of the protein compositions of ...

1990-06-05

479

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

Twenty-two landrace-derived inbred lines from the Spanish Barley Core Collection (SBCC) were found to display high levels of resistance to a panel of 27 isolates of the fungus Blumeria graminis that exhibit a wide variety of virulences. Among these lines, SBCC145 showed high overall resistance and a distinctive spectrum of resistance compared with the other lines. Against this background, the main goal of the present work was to investigate the genetic basis underlying such resistance using a doubled haploid population derived from a cross between SBCC145 and the elite spring cultivar Beatrix. The population was genotyped with the 1,536-SNP Illumina GoldenGate Oligonucleotide Pool Assay (Barley OPA-1 or BOPA1 for short), whereas phenotypic analysis was performed using two B. graminis isolates. A major quantitative trait locus (QTL) for resistance to both isolates was identified on the long arm of chromosome 6H (6HL) and accounted for ca. 60% of the phenotypic ...

2011-07-08

480

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

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

1990-02-26

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

481

New tools for human fat cell alpha-2A adrenoceptor characterization. Identification on membranes and on intact cells using the new antagonist (3H)RX821002

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

1991-04-21

482

Modification of adenylate cyclase by photoaffinity analogs of forskolin

The pharmacology of the alpha-2 adrenoceptor of the human adipocyte was improved by using some new alpha-2 antagonists from different chemical families (imidazolines, benzazepines and benzofuroquinolizines) in biological and binding assays. Moreover, investigations were also carried out to define the binding properties of a new imidazolinic antagonist, RX821002 (2-(2-methoxy-1,4-benzodioxan-2yl)-2-imidazoline), which could be a potential radioligand. (3H)RX821002 binding was very rapid and reversible. Saturation isotherms indicated that (3H)RX821002 labeled, with high affinity, a homogeneous population of noninteracting binding sites with a mean Kd of 0.98 +/- 0.05 nM (n = 6). The binding of (3H)RX821002 on the human fat cell alpha-2 adrenoceptor displayed a specificity which is strictly similar to that obtained with (3H)rauwolscine and which is classical for an alpha-2 A adrenoceptor. The binding parameters of (3H)RX821002 were compared with those obtained with ...

1990-01-01

483

Inhibitory effect of transforming growth factor-. beta. (TGF-. beta. ) on insulin-like growth factor 1 (IGF-1)-induced proliferation and differentiation in primary cultures of pig preadipocytes

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

1989-01-01

484

Inhibitory effect of transforming growth factor-#beta# (TGF-#beta#) on insulin-like growth factor 1 (IGF-1)-induced proliferation and differentiation in primary cultures of pig preadipocytes

The influence of serum, IGF-1 and TGF-{beta} on the differentiation of preadipocytes was examined in primary cultures of porcine adipose tissue cells. In serum-supplemented or serum-free, IGF-1 (1 and 10 nM) had no effect on total cell number. However, IGF-1 (10nM) increased adipocyte number only in serum-supplemented (1% pig serum) cultures, whereas TGF-{beta} (15 pm) reduced the adipocyte number in the presence and absence of IGF-1. Replication of preadipocytes was analyzed with a ({sup 3}H) thymidine assay. Preadipocyte proliferation (cpm in adipocyte fraction) was increased by IGF-1 (10nM) only in cultures containing pig serum. TGF-{beta} had no effect on preadipocyte proliferation specifically, but slightly increased total ({sup 3}H) thymidine incorporation in cultures with serum. Glycerol phosphate dehydrogenase (GPDH) specific activity was decreased by adding TGF-{beta} to serum-free cultures but TGF-{beta} had little effect in serum-supplemented cultures. ...

1990-02-26

485

Increased expression after x-irradiation of MUC1 in cultured human colon carcinoma HT-29 cells

The influence of serum, IGF-1 and TGF-#beta# on the differentiation of preadipocytes was examined in primary cultures of porcine adipose tissue cells. In serum-supplemented or serum-free, IGF-1 (1 and 10 nM) had no effect on total cell number. However, IGF-1 (10nM) increased adipocyte number only in serum-supplemented (1% pig serum) cultures, whereas TGF-#beta# (15 pm) reduced the adipocyte number in the presence and absence of IGF-1. Replication of preadipocytes was analyzed with a ["3H] thymidine assay. Preadipocyte proliferation (cpm in adipocyte fraction) was increased by IGF-1 (10nM) only in cultures containing pig serum. TGF-#beta# had no effect on preadipocyte proliferation specifically, but slightly increased total ["3H] thymidine incorporation in cultures with serum. Glycerol phosphate dehydrogenase (GPDH) specific activity was decreased by adding TGF-#beta# to serum-free cultures but TGF-#beta# had little effect in serum-supplemented cultures. Cellular ...

1991-04-21

486

IL-4/Stat6 activities correlate with apoptosis and metastasis in colon cancer cells

The effect of X-irradiation on production of MUC1 was studied with human colon carcinoma HT-29 cells. As evaluated by immunocytochemical staining, the percentages or MUC1-positive cells in cells at 4 days after 6 Gy irradiation and in unirradiated control cells were 52#+-#3.5% (n=6) and 26#+-#2.8% (n=6), respectively. Flow-cytometric analysis of living cells showed that MUC1 began to rise from day 1, reaching a plateau by day 4 after 6 Gy irradiation. Western blot analysis with monoclonal antibody MY.1E12 against glycosylated MUC1 (mature form) showed dose-dependent increases of two bands (500 and 390 kDa) corresponding to two polymorphic MUC1 alleles. Premature forms of MUC1 (350 and 240 kDa) were detectable with monoclonal antibody HMFG-2 only in irradiated cells, suggesting that new core protein synthesis had been induced. The transcriptional activity of the MUC1 gene was analyzed in terms of transient expression of MUC1-CAT reporter plasmids containing 5'-flanking sequences of the ...

2000-03-01

487

High-affinity lead binding proteins in rat kidney cytosol mediate cell-free nuclear translocation of lead

IL-4-induced Stat6 signaling is active in a variety of cell types and plays a role in cell proliferation/growth and resistance to apoptosis. Using EMSA, we identified differential IL-4/Stat6 activities in colorectal cancer cell lines, HT-29 being active Stat6"h"i"g"h phenotype and Caco-2 being defective Stat6"n"u"l"l phenotype, respectively. Active Stat6"h"i"g"h HT-29 cells exhibited resistance to apoptosis by flowcytometry and aggressive metastasis by Transwell assay compared with defective Stat6"n"u"l"l Caco-2 cells. Comparing one another using RT-PCR, Stat6"h"i"g"h HT-29 cells expressed more mRNA of anti-apoptotic and pro-metastatic genes Survivin, MDM2, and TMPRSS4, while Stat6"n"u"l"l Caco-2 cells expressed more mRNA of pro-apoptotic and anti-metastatic genes BAX, CAV1, and P53, respectively. This is the first study describing correlations of IL-4/Stat6 activities with apoptosis and metastasis in colon cancer. These findings, together with the observation of ...

2008-05-02

488

High-affinity lead binding proteins in rat kidney cytosol mediate cell-free nuclear translocation of lead

The PbII binding characteristics of the previously reported PbII binding proteins of rat kidney cytosol were investigated further. Saturation and Scatchard analysis of /sup 203/Pb binding in whole cytosol and in 40% saturated ammonium sulfate precipitated fractions disclosed a class of relatively high-affinity sites with an apparent Kd of approximately 50 nM and binding capacities of approximately 41 and 9 pmol/mg of protein, respectively. Two /sup 203/Pb binding proteins with approximate molecular masses of 63K and 11.5K daltons and a high molecular weight component (greater than 200K) were isolated by Sepharose-6B column chromatography. The time course of association of /sup 203/Pb with cytosol and the 63K protein showed maximum binding at 18 hr which was stable up to 25 hr at 4 degrees C. The approximate half-time dissociation rate (T 1/2) of specifically bound /sup 203/Pb to the 63K protein was 100 min at 4 degrees C whereas the 11.5K protein showed little dissociation of ...

1985-02-01

489

High extracellular calcium attenuates adipogenesis in 3T3-L1 preadipocytes

The PbII binding characteristics of the previously reported PbII binding proteins of rat kidney cytosol were investigated further. Saturation and Scatchard analysis of "2"0"3Pb binding in whole cytosol and in 40% saturated ammonium sulfate precipitated fractions disclosed a class of relatively high-affinity sites with an apparent Kd of approximately 50 nM and binding capacities of approximately 41 and 9 pmol/mg of protein, respectively. Two "2"0"3Pb binding proteins with approximate molecular masses of 63K and 11.5K daltons and a high molecular weight component (greater than 200K) were isolated by Sepharose-6B column chromatography. The time course of association of "2"0"3Pb with cytosol and the 63K protein showed maximum binding at 18 hr which was stable up to 25 hr at 4 degrees C. The approximate half-time dissociation rate (T 1/2) of specifically bound "2"0"3Pb to the 63K protein was 100 min at 4 degrees C whereas the 11.5K protein showed little dissociation of specifically bound ...

490

Heat Shock Protein 27-Targeted Heptapeptide of the PKC? Catalytic V5 Region Sensitizes Tumors With Radio- and Chemoresistance

We studied the effect of extracellular Ca"2"+ concentration ([Ca"2"+]_e) on adipocyte differentiation. Preadipocytes exposed to continuous [Ca"2"+]_e higher than 2.5 mmol/l accumulated little or no cytoplasmic lipid compared to controls in 1.8 mmol/l [Ca"2"+]_e. Differentiation was monitored by Oil Red O staining of cytoplasmic lipid and triglyceride assay of accumulated lipid, by RT-PCR analysis of adipogenic markers, and by the activity of glycerol-3-phosphate dehydrogenase (GPDH). Elevated [Ca"2"+]_e inhibited expression of peroxisome proliferator-activated receptor #gamma#, CCAAT/enhancer binding protein #alpha#, and steroid regulatory binding element protein. High [Ca"2"+]_e significantly inhibited differentiation marker expression including adipocyte fatty acid binding protein, and GPDH. The decrease in Pref-1 expression that accompanied differentiation also was prevented by high [Ca"2"+]_e. Treatment of 3T3-L1 cells with high [Ca"2"+]_e did not significantly ...

2004-12-10

491

Growth hormone-like factor produced by the tapeworm, Spirometra mansonoides, displaces human growth hormone (hGH) from its receptors on cultured human lymphocytes

Purpose: Previous data suggest that the PKC? catalytic V5 (PKC?-V5) heptapeptide (HEPT) (FEQFLDI) binds HSP27 and blocks HSP27-mediated radio- or chemoresistance. Here we investigated further the in vivo function of the PKC?-V5 HEPT. Methods and Materials: Labeling of HEPT with Cy5.5 or fluorescein isothiocyanate was performed to evaluate in vitro or in vivo distribution of HEPT. A clonogenic survival assay, flow cytometry, and Western blotting of cleaved caspase-3 were performed to determine in vitro sensitization effects of HEPT plus ionizing radiation (IR) versus IR alone or those of HEPT plus cisplatin(Cis) versus Cis alone. A nude mouse xenografting system was also applied to detect in vivo sensitizing effects of HEPT. Results: HEPT efficiently bound to HSP27 and showed sensitization after combined treatment with IR versus treatment with Cis alone in NCI-H1299 lung carcinoma cells, with higher HSP27 expression, which was similar to that of combined treatment ...

2011-05-01

492

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

An analogue of hGH isolated from plerocercoids of the tapeworm Spirometra mansonoides displaces ["1"2"5I]hGH from its receptors in rabbit, rat, and hamster liver membranes. Biologically, plerocercoid growth factor (PGF) is more similar to hGH than to other mammalian GH's but has not been shown to bond human cells. Receptors specific for hGH have been described on cultured human lymphocytes (IM-9). In this study, the authors compared the binding of PGF and hGH in IM-9 cells and in rabbit hepatic membranes. IM-9 lymphocytes (12 x 10"6 cells/tube) were incubated with ["1"2"5I]hGH and increasing concentrations of hGH (ng/ml) or PGF (serial dilutions) for 90 min at 30"0 C. Specific binding (B_0 - NSB) was determined for each dose of hGH or PGF and the binding curves were analyzed by logit-log regression. The results show that PGF displaced ["1"2"5I]hGH from human cells in a dose dependent manner (r = 0.98). Based on the IM-9 assay, 1 ml of the PGF had an activity ...

1986-04-13

493

Feasibility of fissile mass assay of spent nuclear fuel using {sup 252}Cf-source-driven frequency-analysis

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

1987-02-01

494

Continuous detoxification, transformation, and degradation of nitrophenols in upflow anaerobic sludge blanket (UASB) reactors

The feasibility was evaluated using MCNP-DSP, an analog Monte Carlo transport cod to simulate source-driven measurements. Models of an isolated Westinghouse 17x17 PWR fuel assembly in a 1500-ppM borated water storage pool were used. In the models, the fuel burnup profile was represented using seven axial burnup zones, each with isotopics estimated by the PDQ code. Four different fuel assemblies with average burnups from fresh to 32 GWd/MTU were modeled and analyzed. Analysis of the fuel assemblies was simulated by inducing fission in the fuel using a {sup 252}Cf source adjacent to the assembly and correlating source fissions with the response of a bank of {sup 3}He detectors adjacent to the assembly opposite the source. This analysis was performed at 7 different axial positions on each of the 4 assemblies, and the source-detector cross-spectrum signature was calculated for each of these 28 simulated measurements. The magnitude of the cross-spectrum signature follows a smooth upward ...

1996-10-01

495

Biphasic calcium sulfate dihydrate/iron-modified alpha-tricalcium phosphate bone cement for spinal applications: in vitro study

The anaerobic transformation and degradation of nitrophenols by granular sludge was investigated in upflow anaerobic sludge blanket (UASB) reactors continuously fed with a volatile fatty acid (VFA) mixture as the primary substrate. During the start-up, subtoxic concentrations of 2-nitrophenol (2-NP), 4-nitrophenol (4-NP), and 2, 4-dinitrophenol (2, 4-DNP) were utilized. 4-NP and 2, 4-DNP were readily converted to the corresponding aromatic amine; whereas 2-NP was converted to nonaromatic products via intermediate formation of 2-aminophenol (2-AP). These conversions led to a dramatic detoxification of the mononitrophenols because the reactors treated the nitrophenolics at the concentrations which were over 25 times higher than those that caused severe inhibition. VFA removal efficiencies greater than 99% were achieved in both reactors at loading rates greater than 11.4 g COD per liter of reactor volume per day even at volumetric loading of mononitrophenols up to 910 mg/L {center_dot} d. ...

1996-08-20

496

Assessment of chromosomal abnormalities in sperm of infertile men using sperm karyotyping and multicolour fluorescence in situ hybridization (FISH)

In this study, the cytocompatibility of new 'iron-modified/alpha-tricalcium phosphate (IM/alpha-TCP) and calcium sulfate dihydrate (CSD)' bone cement (IM/alpha-TCP/CSD-BC) intended for spinal applications has been approached. The objective was to investigate by direct-contact osteoblast-like cell cultures (from 1 to 14 days) the in vitro cell adhesion, proliferation, morphology and cytoskeleton organization of MG-63 cells seeded onto the new cements. The results were as follows: (a) quantitative MTT-assay and scanning electron microscopy (SEM) showed that cell adhesion, proliferation and viability were not affected with time by the presence of iron in the cements; (b) double immunofluorescent labeling of F-actin and alpha-tubulin showed a dynamic interaction between the cell and its porous substrates sustaining the locomotion phenomenon on the cements' surface, which favored the colonization, and confirming the biocompatibility of the ...

2010-04-15

497