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At least 721 records · Page 40Linked to original sources
PubMed1988

Comparison by restriction fragment pattern analysis and molecular characterization of some European isolates of Suid herpesvirus 1: a contribution to strain differentiation of European isolates.

Eleven European isolates of Suid herpesvirus type 1 (SHV-1) were compared by restriction fragment pattern analyses and Southern blot hybridization using different genomic probes. The presence of strain discriminative 4 major genome types and several subtypes as well as the molecular distinctions between these were demonstrated. Evolutionary relationships between the different types are suggested on the basis of homologous restriction fragment length polymorphisms.

Blotting, Southern↗
PubMed1978

Studies on isolated subcellular components of cat pancreas. III. Alanine-sodium cotransport in isolated plasma membrane vesicles.

Transport of alanine was studied in isolated plasma membrane vesicles from cat pancreas using a rapid filtration technique. The uptake is osmotically sensitive and the kinetics of L-alanine transport are biphasic showing a saturable and a nonsaturable component. The saturable component is seen only when a sodium gradient directed from the medium to the vesicular space is present. Under this condition an overshooting uptake of L-but not of D-alanine occurs. The Na+ gradient stimulated uptake of L-alanine is inhibited by L-serine and L-leucine and stimulated when the membrane vesicles had been preloaded with L-alanine, L-serine or L-leucine. The ionophore monensin inhibits stimulation of uptake caused by a sodium gradient. In the presence of valinomycin or carbonyl cyanide p-trifluoromethoxyphenylhydrazone (CFCCP), the sodium-dependent transport is augmented in vesicles preloaded with K2SO4 or H+ ions (intravesicular pH 5.5), respectively. In the presence of different anions, the Na+-dependent transport is stimulated according to increasing anionic penetration through membrane (lipid solubility). We conclude that a sodium dedpendent electrogenic amino acid transport system is present in pancreatic plasma membranes.

Alanine↗
PubMed1993

A cobblestone cell isolated from the human omentum: the mesothelial cell; isolation, identification, and growth characteristics.

Normal human mesothelial cells (NHMC) were isolated from pieces of human omentum. The cell yield was approximately one million cells per square centimeter omentum. The mesothelial cells were identified by their positive staining with monoclonal antibodies against cytokeratins 6 and 18. Transmission electronmicroscopy of cultured NHMC revealed many microvilli on the apical surface and many mitochondria and pinocytotic vesicles in the cytoplasm, indicating active transmembrane transport. Growth of NHMC was directly related to the concentration of human serum or of fetal bovine serum in the growth medium. Addition of epidermal growth factor with or without hydrocortisone resulted in a significant increase of NHMC growth; when endothelial cell growth factor, insulin, or hydrocortisone were added no such increase was observed. Seeding NHMC at densities less than 3000/cm2 did not result in monolayer formation. The mesothelial cells were serially passed in growth medium M199 with added 10% fetal bovine serum up to 7 passages. However, after Passage 4 the cells changed into giant cells with an irregular pattern, and a lack of intracellular cytokeratin expression was observed for most of the cells.

Antibodies, Monoclonal↗
PubMed2004

Isolation and characterization of dibenzofuran-degrading Comamonas sp. strains isolated from white clover roots.

Three dibenzofuran (DF)-degrading strains were newly isolated from roots of white clover ( Trifolium repens L.) and poplar trees grown in DF-contaminated soil samples. These strains, designated KD2, KD7, and PD1, were characterized as Comamonas sp. on the basis of their 16S rDNA sequences and physiological characteristics. The metabolites produced when strain KD7 was incubated with DF were identified by gas chromatography-mass spectrometry (GC-MS) analysis. Interestingly, strain KD7 was found to have two pathways for DF degradation, beginning with angular dioxygenation at carbons 4 and 4a, and lateral dioxygenation at carbons 1 and 2, respectively. Furthermore, strains KD2 and KD7 not only achieved efficient root colonization in clover but also promoted clover growth. They are the first reported Comamonas sp. strains capable of utilizing DF as a sole carbon source. This provides additional information on the diversity of DF-degrading bacteria.

Benzofurans↗
PubMed2026

Pseudaquabacterium prasiolae sp. nov., Isolated from the Freshwater Green Alga Prasiola japonica, and Rubrivivax soli sp. nov., Isolated from Soil, with Reclassification of Aquabacterium humicola as Pseudaquabacterium humicola comb. nov.

Two Gram-stain-negative, catalase- and oxidase-positive, strictly aerobic, non-flagellated rod-shaped bacteria, designated OR-4T and RP6-9T, were isolated from the freshwater green alga Prasiola japonica and soil in Republic of Korea, respectively. Strain OR-4T exhibited gliding motility, whereas strain RP6-9T lacked gliding motility. Strain OR-4T grew at 10-30 °C, pH 6.0-9.0, and 0-1.5% (w/v) NaCl, while strain RP6-9T grew at 20-35 °C, pH 6.0-9.0, and 0-1.0% (w/v) NaCl. Both strains contained ubiquinone-8 as the sole respiratory quinone and phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol as major polar lipids; strain OR-4T additionally possessed an unidentified phospholipid and an unidentified polar lipid. The predominant fatty acids of OR-4T were C12:0, C16:0, summed feature 3 (C16:1ω6c and/or C16:1ω7c), and summed feature 8 (C18:1ω7c and/or C18:1ω6c), whereas RP6-9T contained C12:0, C16:0, and summed feature 3 as major components. The genomic DNA G + C content of both strains was 71.0 mol%. Whole-genome-based phylogenomic analyses placed OR-4T and RP6-9T within the genera Pseudaquabacterium and Rubrivivax, respectively, forming distinct lineages. Comparative analyses of average nucleotide identity, digital DNA-DNA hybridization, and average amino acid identity further supported their assignment to these genera while confirming their separation from previously described species. Based on combined phenotypic, chemotaxonomic, and genomic evidence, strains OR-4T and RP6-9T represent novel species, for which the names Pseudaquabacterium prasiolae sp. nov. (type strain OR-4T =KACC 22752T =NBRC 116024T) and Rubrivivax soli sp. nov. (type strain RP6-9T =KACC 24055T =DSM 119932T) are proposed. Phylogenomic analyses also support the reclassification of Aquabacterium humicola as Pseudaquabacterium humicola comb. nov. (type strain RJY3T =KCTC 92105T =NBRC 115831T).

Phylogeny↗
PubMed2004

Genomic sequence of an isolate of Newcastle disease virus isolated from an outbreak in geese: a novel six nucleotide insertion in the non-coding region of the nucleoprotein gene. Brief Report.

We have completely sequenced the genome of an isolate of Newcastle disease virus (NDV) obtained following an outbreak in geese. The genomic sequence consists of 15192 nt, which is six nt longer than the published full length genome of the NDV strains La Sota, B1, Clone-30, Beaudette C and HB V4. The six nt insertion was located in the non-coding region of the nucleoprotein (NP) gene between nt 1646 and nt 1647 of the NDV genome (numbered according to the genomic sequence of the La Sota strain). An additional 22 NDV strains were searched for the existence of this six nt insertion. NDVs in genotypes VI, VII, VIII and IX had this insertion while NDV's in genotypes I, II, III, IV, and V did not. The significance of this insertion in NDV evolution is discussed.

Animals↗
PubMed1999

Taxonomic studies of extremely barophilic bacteria isolated from the Mariana Trench and description of Moritella yayanosii sp. nov., a new barophilic bacterial isolate.

We have isolated two strains of extremely barophilic bacteria from sediment collected from the world's deepest ocean floor in the Mariana Trench, Challenger Deep, at a depth of 10898m [Kato C, Li L, Nogi Y, Nakamura Y, Tamaoka J, Horikoshi K (1998) Appl Environ Microbiol 64:1510-1513]. One strain, DB21MT-2, was identified as a strain of Shewanella benthica, and the other strain, DB21MT-5, is closely affiliated with members of the genus Moritella on the basis of 16S rDNA sequence analysis. The hybridization values for DNA-DNA relatedness between DB21MT-5 and the Moritella reference strains were significantly lower than that accepted as the phylogenetic definition of a species. Based on this and other taxonomic differences, strain DB21MT-5 appears to represent a novel obligately barophilic deep-sea Moritella species. The name Moritella yaynanosii (JCM 10263) is proposed. This is the first proposed species of obligately barophilic bacteria of the genus Moritella.

Bacteria↗
PubMed2005

Isolation and metabolite production by Penicillium roqueforti, P. paneum and P. crustosum isolated in Canada.

Penicillium roqueforti, P. crustosum and P. paneum grow on ensiled grain and recycled feed unless properly treated. The former two species occur also on cut lumber in Canada. These are known to produce a number of secondary metabolites including roquefortine. In cooler dairy production areas, including Scandinavia and North America, cattle toxicosis has been associated with silage contaminated by these fungi. We collected strains associated with cow or cattle toxicoses. The principal metabolites were determined making use of a new extraction method and analysis combining HPLC, LC/MS/MS, and LC/NMR. Penicillium roqueforti and P. crustosum required amino acid nitrogen for metabolite formation and their toxins were formed under conditions of low oxygen (20-30% saturation). Production of roquefortine C occurred on depletion of the available nitrogen and penitrem A on depletion of carbon source. Yield was reduced by excess carbon. Medium osmotic tension (aw) affected metabolite production by the two species differently. Penicillium paneum was associated with ill-thrift of dairy cows and P. roqueforti was associated with more serious symptoms. Our data suggest a physiological basis for the common occurrence of roquefortine C in silage without serious consequences and the alternative, the presence of roquefortine C and toxicoses. The strain isolated from lumber was the best producer of the toxins studied. This is the first report of the toxigenic potential of P. roqueforti and P. paneum from Canada.

Animals↗
PubMed1984

Apolipoprotein E Bethesda. Isolation and partial characterization of a variant of human apolipoprotein E isolated from very low density lipoproteins.

A variant of apolipoprotein E, denoted E Bethesda, has been identified in the plasma of a 72-year-old woman with type III hyperlipoproteinemia. An offspring of the proband also has this variant and type III hyperlipoproteinemia. Apolipoprotein E Bethesda was isolated by preparative isoelectrofocusing followed by preparative SDS-polyacrylamide gel electrophoresis from the very low density lipoproteins of the proband's son. The purity and the identity of the preparation were analyzed by analytical SDS-polyacrylamide gel electrophoresis, two-dimensional gel electrophoresis and by immunochemical analysis. Apolipoprotein E Bethesda migrates in the E 1 position and its electrophoretic mobility is not affected by neuraminidase treatment. The protein is shifted to the E3 position after cysteamine treatment. The amino acid composition revealed the presence of two cysteine residues. These data support the concept that the apolipoprotein E Bethesda allele is derived from a mutation of the E2 or E2* allele.

Aged↗
PubMed1979

Astroglial and axonal proteins in isolated brain filaments. I. Isolation of the glial fibrillary acidic protein and of an immunologically active cyanogen bromide peptide from brain filament preparations of bovine white matter.

The glial fibrillary acidic protein and an immunologically active cyanogen bromide peptide were purified by immunoaffinity chromatography from 8 M urea extracts of brain filament preparations isolated from bovine white matter according to Norton's procedure. The protein accounted for approximately 30% of the total protein in this preparation and for the largest fraction in the 50 000 molecular weight range. The fraction not absorbed to the immuno-Sepharose column reacted with neurofilament antisera by double immunodiffusion. On sodium dodecyl sulfate gel electrophoresis the main bands in the non-adsorbed fraction were at 74 000 daltons and above 100 000. Several bands were seen in the 50 000 molecular weight range. It is concluded that glio- and neurofilaments co-purify together in Norton's procedure and that neurofilaments are probably heterogeneous in polypeptide composition.

Animals↗
PubMed1980

Astroglial and axonal proteins in isolated brain filaments. II. Isolation of a 70 000-dalton polypeptide from bovine brain filament preparations by immunoaffinity chromatography with antineurofilament antisera.

Antineurofilament antisera raised against chicken brain antigen were used to isolate neurofilament protein from bovine brain filament preparations by immunoaffinity chromatography. Glial fibrillary acidic protein, the subunit of glial filaments, passed unabsorbed through the column. The Mr 70 000 component of the neurofilament triplet was tightly absorbed to the column and essentially the only protein eluted at pH 2.5. The other two components of the triplet, of approximately 200 000 and 150 000 daltons, were less tightly attached to the column and eluted at pH 6.0 in 5 M urea.

Animals↗
PubMed1979

The isolation and lipid composition of subcellular fractions derived from neuronal perikarya isolated in bulk from rabbit cerebral cortex.

(1) Neuronal perikarya were isolated from rabbit cerebral cortex and were homogenized and separated into a number of subcellular membrane fractions using differential and discontinuous density gradient centrifugation. (2) The efficiency of homogenizing the nerve cell bodies could be greatly increased by the preliminary passage of the cell body fraction through a micropipet tip. (3) Of a number of density media, a discontinuous gradient of metrizamide gave the best resolution of the mitochondrial and lysosomal marker enzymes found in the crude mitochondrial fraction. This yielded a purified mitochondrial fraction and several lighter membrane fractions. (4) The lighter membrane fractions in metrizamide contained a degree of mitochondrial contamination which could be removed by a second gradient spin on 1.2 M sucrose, producing a second mitochondrial fraction and two light membrane fractions. (5) Based on marker analyses, fractions enriched in nuclei, mitochondria, microsomes and plasma membrane/lysosomes were produced. (6) The two mitochondrial fractions showed the highest, and the nuclear fraction the lowest specific phospholipid content. Cholesterol: phospholipid molar ratios showed a gradient of values from a low (0.2) for the mitochondria and nuclei to an intermediate value (0.4) for the microsomes to a high (0.6) for the light membrane fractions. (7) Phospholipid distributions indicated that for the nuclear and mitochondrial fractions phosphatidylinositol was more abundant than sphingomyelin or phosphatidylserine, while for the microsomes and the two light membrane fractions these three phospholipids were present in almost equal amounts.

Acetylglucosaminidase↗
PubMed1982

Isolation of glucagon-37 (bioactive enteroglucagon/oxyntomodulin) from porcine jejuno-ileum. Isolation of the peptide.

A 37 amino acid-peptide has been isolated from porcine jejuno-ileum on the basis of its glucagon-like activity in liver (interaction with glucagon-binding sites and activation of adenylate cyclase) using gel filtration, ion-exchange and high-performance liquid chromatography. Depending on the criteria chosen, this peptide is referred to as either 'bioactive enteroglucagon' (activity in liver), 'oxyntomodulin' (specific action in gastric oxyntic glands) or 'glucagon-37' (chemical structure).

Adenylyl Cyclases↗
PubMed1993

Isolation and characterization of a high molecular weight type IV collagenase isolated from human carcinoma tissue.

A proform of high molecular weight type IV collagenase was isolated and purified 1230-fold from human metastatic carcinoma tissue. Like matrix metalloproteinases (MMPs), the enzyme was activated by trypsin and degraded type IV collagen and gelatin at a neutral pH, the activity was inhibited by EDTA and o-phenanthroline. However, the molecular weight was much higher than MMPs which degraded type IV collagen, gelatinase A (MMP-2; 72 kDa gelatinase/type IV collagenase) (EC 3.4.24.24), gelatinase B (MMP-9; 92 kDa gelatinase/type IV collagenase) (EC 3.4.24.35), stromelysin-1 (MMP-3; 57 kDa) (EC 3.4.24.17) and stromelysin-2 (MMP-10; 57 kDa) (EC 3.4.24.22). The other significant difference from MMPs was that the enzyme was not activated by 4-aminophenylmercuric acetate nor inhibited by TIMP. Taking together these results, this high molecular weight type IV collagenase might be a newly found enzyme different from MMPs or might have the same configuration as MMPs already reported.

Collagen↗
PubMed1982

Factors which can influence beta-oxidation by peroxisomes isolated from livers of clofibrate treated rats. Some properties of peroxisomal fractions isolated in a self-generated Percoll gradient by vertical rotor centrifugation.

1. Oxidation of radioactivity labeled acyl-CoA esters by apparently intact peroxisomes, isolated from rats treated with clofibrate, was stimulated by about 100% by added oxaloacetate. Added pyruvate gave much smaller stimulation. This stimulation is proposed to be due to re-oxidation of NADH generated during peroxisomal beta-oxidation of acyl-CoA esters. 2. Additions of these alpha-ketoacids, and of CoA, were found to influence extent of peroxisomal beta-oxidation of [U-14C]palmitoyl-CoA. Various concentrations of added CoA was markedly less inhibitory to oxidation of [14-14C]erucoyl-CoA and [10-14C]elaidoyl-CoA, as compared to [U-14C]palmitoyl-CoA. 3. Added NADP+, at concentrations higher than 0.5 mM, added NADPH and NADH were inhibitory to peroxisomal beta-oxidation of [U-14C]palmitoyl-CoA. Significant inhibition was found at concentrations which were too high for this to be of significance in vivo.

Animals↗
PubMed1988

Oxyntomodulin (glicentin-(33-69)): pharmacokinetics, binding to liver cell membranes, effects on isolated perfused pig pancreas, and secretion from isolated perfused lower small intestine of pigs.

The pharmacokinetics of purified synthetic oxyntomodulin were studied after infusing it into euglycaemic pigs at two rates. The elimination of the peptide from plasma was characterized by two components, a fast one (t1/2 7.2 +/- 0.6 min) and a slow one (t1/2 20.4 +/- 3.8 min) (mean +/- S.E.M., n = 7). The metabolic clearance rate was independent of infusion rate (6.96 +/- 0.99 vs 7.44 +/- 0.98 ml/kg . min (mean +/- S.E.M., n = 7). The synthetic peptide bound to pig hepatic glucagon receptors, but with approximately 2% of the affinity of glucagon, and showed insulinotropic and somatostatinotropic effects when infused into isolated perfused pig pancreases at concentrations higher than 10(-10) M. A dose-dependent increase was also shown for pancreatic glucagon output. A naturally occurring peptide, identified as oxyntomodulin by gel filtration and HPLC, was released into the circulation from the pig lower small intestinal mucosa upon intraluminal administration of glucose, and represented 25 +/- 3.8% of the secreted glucagon-like immunoreactivity. 11 +/- 2.3% of the secreted glucagon-like immunoreactivity was indistinguishable from glucagon itself upon gel filtration; thus at least 36% of the glucagon-like immunoreactivity secreted from the intestinal mucosa is already in an active form.

Animals↗