| homogeneous bioluminescence competitive binding assay for folate based on a coupled glucose-6-phosphate dehydrogenase--bacterial luciferase enzyme system. | a homogeneous bioluminescence competitive binding assay for folate was developed by using a coupled enzyme system of glucose-6-phosphate dehydrogenase (g6pdh) and bacterial luciferase. a highly substituted g6pdh-folate conjugate was prepared by employing an n-hydroxysuccinimide/carbodiimide method. folate binding protein inhibits the activity of the conjugate. in the presence of folate, there is a competition between folate and the g6pdh-folate conjugate for the binding site of the folate bindin ... | 1996 | 8815749 |
| analysis of a pressure-regulated operon from the barophilic bacterium strain db6705. | a pressure-regulated operon was cloned and sequenced from a barophilic bacterium strain db6705. gene expression by this promote was regulated at the level of transcription by elevated pressure in the barophilic strain. the promoter sequence was similar to the promoter of omph from photobacterium sp. strain ss9. | 1996 | 8824840 |
| purification and characterization of a marine bacterial beta-galactoside alpha 2,6-sialyltransferase from photobacterium damsela jt0160. | a bacterial sialyltransferase, named sialyltransferase 0160, was purified from a marine bacterium that had been isolated from seawater from sagami bay, kanagawa. this strain has been identified as photobacterium damsela, and named p. damsela jt0160. sialyltransferase 0160 was purified 688-fold to homogeneity from the crude extract of the cells with a yield of 19% using a combination of anion exchange chromatography, hydroxyapatite chromatography, gel filtration chromatography, and affinity chrom ... | 1996 | 8864851 |
| identification of sodc encoding periplasmic [cu,zn]-superoxide dismutase in salmonella. | sodc, encoding [cu,zn]-cofactored superoxide dismutase, once thought to be virtually confined to eukaryotes, has now been described in many gram-negative pathogens that have their primary niche of colonization in the upper respiratory tract. their role in host-parasite interactive biology is unknown. we here show that members of the major human and animal enteric pathogenic species salmonella harbour a version of sodc most closely resembling that found in brucella abortus. the enzyme it encodes ... | 1996 | 8869506 |
| effect of modified atmosphere packaging on the tvb/tma-producing microflora of cod fillets. | cod fillets (gadus morhua) were packed under modified atmospheres, with four different gas compositions (60% co2-10% o2-30% n2, 60% co2-20% o2-20% n2, 60% co2-30% o2-10% n2, 60% co2-40% o2), and stored at 6 degrees c. plate counts were carried out after 3, 4, 5, 6 and 7 days, to follow the growth of aerobic and anaerobic bacteria, lactic acid bacteria, h2s-producing bacteria and enterobacteriaceae. the production of total volatile bases (tvb) and trimethylamine (tma), and the changes in ph of th ... | 1996 | 8880310 |
| flavin reductase p: structure of a dimeric enzyme that reduces flavin. | we report the structure of an nadph:fmn oxidoreductase (flavin reductase p) that is involved in bioluminescence by providing reduced fmn to luciferase. the 1.8 a crystal structure of flavin reductase p from vibrio harveyi was solved by multiple isomorphous replacement and reveals that the enzyme is a unique dimer of interlocking subunits, with 9352 a2 of surface area buried in the dimer interface. each subunit comprises two domains. the first domain consists of a four-stranded antiparallel beta- ... | 1996 | 8885832 |
| interspecific luciferase beta subunit hybrids between vibrio harveyi, vibrio fischeri and photobacterium leiognathi. | bacterial luciferase (ec 1.14.14.3) is a heterodimer composed of alpha- and beta-chains encoded by luxa and luxb, respectively. although some interspecific combinations of these subunits lead to active enzyme, others do not. the beta subunits of vibrio fischeri and photobacterium leiognathi form active enzyme with the alpha subunits of v.fischeri, p.leiognathi and vibrio harveyi, while the beta subunit from v.harveyi only complements the alpha subunit of v.harveyi. inactivity is caused by a lack ... | 1996 | 8888147 |
| lumazine protein and the excitation mechanism in bacterial bioluminescence. | the spectral properties of lumazine protein and mixtures with the intermediates of the bacterial luciferase reaction, are reviewed. measurements of fluorescence dynamics in particular have been employed with the aim of elucidating the mechanism by which lumazine protein functions in the bioluminescence of the bacteria of the type photobacterium. the reaction of bacterial luciferase with its substrates produces bioluminescence emission with a spectral maximum at 496 nm. this spectrum is the same ... | 1993 | 8298053 |
| novel dimeric interface and electrostatic recognition in bacterial cu,zn superoxide dismutase. | eukaryotic cu,zn superoxide dismutases (cuznsods) are antioxidant enzymes remarkable for their unusually stable beta-barrel fold and dimer assembly, diffusion-limited catalysis, and electrostatic guidance of their free radical substrate. point mutations of cuznsod cause the fatal human neurodegenerative disease amyotrophic lateral sclerosis. we determined and analyzed the first crystallographic structure (to our knowledge) for cuznsod from a prokaryote, photobacterium leiognathi, a luminescent s ... | 1996 | 8917495 |
| toxicological evaluation of harmful substances by in situ enzymatic and biological detection in high-performance thin-layer chromatography. | the efficiency of a chromatographic analysis method is determined by the selectivity of the chromatographic separation and the specificity of the detection method. in the case of high-performance thin-layer chromatography (hptlc) the separated components can be detected and quantified directly on the chromatogram by physical and chemical methods. by coupling high-performance thin-layer chromatography with biological or biochemical inhibition tests it was possible to detect toxcologically active ... | 1996 | 8938396 |
| nucleotide sequence and functional analysis of the luxe gene encoding acyl-protein synthetase of the lux operon from photobacterium leiognathi. | nucleotide sequence of the luxe gene genbank accession no. u66407 from photobacterium leiognathi pl741 has been determined, and the amino acid sequence of acyl-protein synthetase encoded by the luxe gene is deduced. nucleotide sequence reveals that the luxe gene encodes acyl-protein synthetase, which is a component of the fatty acid reductase complex that is responsible for converting fatty acid to aldehyde as substrate in the luciferase-catalyzed bioluminescence reaction. the acyl-protein synth ... | 1996 | 8941351 |
| qsar study of the toxicity of nitrobenzenes to river bacteria and photobacterium phosphoreum. | | 1997 | 8952935 |
| a bioluminescent gamma-aminobutyrate assay for monitoring its release from inhibitory nerve endings. | a bioluminescent gaba assay is described. the principle of the procedure is based on the action of gabase (gaba-aminotransferase plus succinic semialdehyde dehydrogenase), coupled to the detection of succinic semialdehyde and nadh, using photobacterium luciferase. the method was used for monitoring gaba release from depolarized brain slices. | 1996 | 8965088 |
| determination of acute toxicity of polychlorinated biphenyls to photobacterium phosphoreum. | | 1997 | 8975803 |
| antigenicity and n-terminal amino acid sequence of a 35 kda porin-like protein of listonella (vibrio) anguillarum: comparison among different serotypes and other bacterial species. | listonella (vibrio) anguillarum, an important fish pathogen, is divided into 10 serotypes according to o-antigens present on the outer membrane. however, the biochemical and immunological properties of porin proteins have not been reported. in this study, the antigenicity and n-terminal amino acid sequence of the 35 kda porin-like-major outer membrane protein (omp35la) were compared among different serotypes of l. anguillarum as well as other bacteria. in western blotting analysis, antisera agai ... | 1996 | 8987710 |
| cloning, sequencing and overexpression of the gene encoding malate dehydrogenase from the deep-sea bacterium photobacterium species strain ss9. | the gene encoding malate dehydrogenase (mdha) was obtained from the psychrophilic, barophilic, deep-sea isolate photobacterium species strain ss9. the ss9 mdha gene directed high levels of malate dehydrogenase (mdh) production in escherichia coli. a comparison of ss9 mdh to three mesophile mdhs, a mdh sequence obtained from another deep-sea bacterium, and to other psychrophile proteins is presented. | 1997 | 9003456 |
| cloning, sequencing, and analysis of a gene cluster from chelatobacter heintzii atcc 29600 encoding nitrilotriacetate monooxygenase and nadh:flavin mononucleotide oxidoreductase. | nitrilotriacetate (nta) is an important chelating agent in detergents and has also been used extensively in processing radionuclides. in chelatobacter heintzii atcc 29600, biodegradation of nta is initiated by nta monooxygenase that oxidizes nta to iminodiacetate and glyoxylate. the nta monooxygenase activity requires two component proteins, component a and component b, but the function of each component is unclear. we have cloned and sequenced a gene cluster encoding components a and b (nmoa an ... | 1997 | 9023192 |
| classification of photobacteria associated with spoilage of fish products by numerical taxonomy and pyrolysis mass spectrometry. | forty strains of luminous and non-luminous photobacterium phosphoreum isolates from cod (gadus morhua) and seven reference strains of psychrotolerant and mesophilic photobacteria were examined for 156 unit characters in a numerical taxonomic study. the fish strains were isolated from the intestines, from spoiled products and by using a specific detection method. the data were analysed using the similarity coefficient and the unweighted pair-group with arithmetic averages algorithm. in addition t ... | 1997 | 9060149 |
| evaluation of the genotoxicity potential (by mutatox test) of ten pesticides found as water pollutants in cyprus. | ten pesticides: aldicarb, aldicard sulfone, aldicarb oxide, carbofuran, propoxur, methomyl, diuron, linuron, alachlor and parathion-methyl, found as water pollutants in cyprus, were evaluated for their genotoxicity potential with the mutatox test, both directly and after exogenous activation with s9 hepatic enzymes. a dark variant (m 169) of the photobacterium phosphoreum was used as the test organism. trials were undertaken in triplicate using ground-water spiked with pesticides solutions at te ... | 1996 | 9111768 |
| effect of river and wetland sediments on toxicity of metolachlor. | metolachlor is a preplant, preemergent herbicide applied to corn and soybean fields. agricultural runoff after application can cause the herbicide to enter into natural waters. the objective of this study was to determine the effect of river and wetland sediments on the toxicity of metolachlor. sediments from a river and a wetland (separately) were mixed with ottawa sand (0, 25, 50, 75, and 100%). metolachlor (in water) and sediment were mixed in an orbital shaker for 8 hr; the mixture was centr ... | 1997 | 9126436 |
| mixture toxicity of nitrobenzene and trinitrobenzene using the marine bacterium vibrio harveyi as the test organism. | vibrio harveyi, a bioluminescent marine bacterium, was used to evaluate combined or mixture toxicity of two organic compounds, nitrobenzene and trinitrobenzene. an estimated median effective concentration (ec50) and confidence interval were determined for each chemical. these chemicals at their ec50 were evaluated in combination and an additive index method was used to determine a numerical toxicology value. combinations at 20% intervals of the ec50 were performed using isopleths. the isopleths ... | 1997 | 9126438 |
| ecotoxicity monitoring--use of vibrio fischeri. | the proliferation of chemical substances having the potential to pollute any environmental medium (air, land, water), or humans via occupational exposure, la considerable. whilst chemical analytical techniques exist for the measurement of some of these chemicals, many of the methods involve costly techniques of considerable sophistication-quantification may be even more difficult. in less developed countries where sophisticated techniques may not be available or supplies of reagents, compressed ... | 1996 | 9127506 |
| cysteine-286 as the site of acylation of the lux-specific fatty acyl-coa reductase. | the channelling of fatty acids into the fatty aldehyde substrate for the bacterial bioluminescence reaction is catalyzed by a fatty acid reductase multienzyme complex, which channels fatty acids through the thioesterase (luxd), synthetase (luxe) and reductase (luxc) components. although all three components can be readily acylated in extracts of different luminescent bacteria, this complex has been successfully purified only from photobacterium phosphoreum and the sites of acylation identified o ... | 1997 | 9128139 |
| light organ symbioses in fishes. | most bioluminescent fishes are self-luminescent, but a substantial minority of bioluminescent teleosts produce light that is due to symbiotic luminous bacteria housed in elaborate light organs. the majority of symbiotically bioluminescent fishes (ten families in five orders) harbors common free-living species of marine luminous bacteria: photobacterium phosphoreum, p. leiognathi, and p. fischeri (= vibrio fischeri). others, associated with the beryciform family anomalopidae and nine families in ... | 1993 | 8305135 |
| purification and characterization of flavoproteins and cytochromes from the yellow bioluminescence marine bacterium vibrio fischeri strain y1. | several flavoproteins and cytochromes that occur as major components in extracts of the yellow bioluminescence y1 strain of the marine bacterium vibrio fischeri have been purified and characterized with respect to their mass (sds/page and matrix-assisted laser-desorption/ionization ms), chromatographic properties, n-terminal sequence, and spectroscopy (absorption, fluorescence emission and anisotropy decay). the investigated proteins were as follows: yellow fluorescence protein (yfp) with bound ... | 1997 | 9183020 |
| spectroscopic characterization of recombinant cu,zn superoxide dismutase from photobacterium leiognathi expressed in escherichia coli: evidence for a novel catalytic copper binding site. | cu,zn superoxide dismutase from photobacterium leiognathi has been cloned and expressed in escherichia coli. the circular dichroism spectrum in the uv region of the recombinant protein indicates an higher content of random coil structure with respect to the eukaryotic enzymes. investigation of the active site by optical, cd, and epr spectroscopy indicates a different coordination geometry around the catalytic copper site with respect to the eukaryotic enzymes. in particular a different orientati ... | 1997 | 9188710 |
| strain characterization and classification of oxyphotobacteria in clone cultures on the basis of 16s rrna sequences from the variable regions v6, v7, and v8. | a major problem in development of a polyphasic taxonomy is that the identification of oxyphotobacterial strains (cyanobacteria and prochlorophytes) in culture collections may be incorrect. we have therefore developed a diagnostic system using the dna sequence polymorphism in the 16s rrna regions v6 to v8 for individual strain characterization and identification. pcr primers amplifying v6 to v8 from oxyphotobacteria in unialgal cultures were constructed. direct solid-phase or cyclic sequencing wa ... | 1997 | 9212409 |
| evaluation of the toxicity of uncaria tomentosa by bioassays in vitro. | aqueous extracts of uncaria tomentosa (willdenow ex roemer and schultes) dc. (rubiaceae) ('uña de gato'), were analyzed for the presence of toxic compounds in chinese hamster ovary cells (cho) and bacterial cells (photobacterium phosphoreum). toxicity was evaluated by four systems: neutral red assay (nr), total protein content (kb), tetrazolium assay (mtt) and microtox test. the extracts of u. tomentosa did not show toxicity in vitro at the concentrations tested. testing in vitro could be a valu ... | 1997 | 9292411 |
| microbiological spoilage of fish and fish products. | spoilage of fresh and lightly preserved fish products is caused by microbial action. this paper reviews the current knowledge in terms of the microbiology of fish and fish products with particular emphasis on identification of specific spoilage bacteria and the qualitative and quantitative biochemical indicators of spoilage. shewanella putrefaciens and pseudomonas spp. are the specific spoilage bacteria of iced fresh fish regardless of the origin of the fish. modified atmosphere stored marine fi ... | 1996 | 8913813 |
| shewanella woodyi sp. nov., an exclusively respiratory luminous bacterium isolated from the alboran sea. | thirty-four strains of nonfermentative, respiratory, luminous bacteria were isolated from samples of squid ink and seawater from depths of 200 to 300 m in the alboran sea. although these strains had a few properties similar to properties of shewanella (alteromonas) hanedai, they did not cluster phenotypically with any previously described bacterium. the nucleotide sequence of a 740-bp segment of luxa was not homologous with other known luxa sequences but clustered with the luxa sequences of shew ... | 1997 | 9336902 |
| the pki gene encoding pyruvate kinase i links to the luxz gene which enhances bioluminescence of the lux operon from photobacterium leiognathi. | partial 3'-end nucleotide sequence of the pki gene (genbank accession no. af019143) from photobacterium leiognathi atcc 25521 has been determined, and the encoded pyruvate kinase i is deduced. pyruvate kinase i is the key enzyme of glycolysis, which converts phosphoenol pyruvate to pyruvate. alignment and comparison of pyruvate kinase is from p. leiognathi, e. coli and salmonella typhimurium show that they are homologous. nucleotide sequence reveals that the pki gene is linked to the luxz gene t ... | 1997 | 9345300 |
| h-ns protein represses transcription of the lux systems of vibrio fischeri and other luminous bacteria cloned into escherichia coli. | high expression in escherichia coli of the lux system cistron of a luminous bacteria under its own control has been accomplished only for the vibrio fischeri lux system at high cell density. mutation of the hns gene in e. coli has resulted in strong expression of the v. fischeri lux system at low cell density even in an rpos-deleted strain of e. coli that emits very low levels of luminescence. the e. coli double mutant, mc4110 hns::kan rpos::tet carrying the lux system of v. fischeri, developed ... | 1997 | 9353217 |
| the toxicity of substituted phenolic compounds to a detoxifying and an acetic acid bacterium. | in the detoxifying bacterium acinetobacter calcoaceticus 69-v and in the acetic acid bacterium acetobacter methanolicus mb 58, glucose and xylose are oxidized, respectively, via pqq-dependent membrane-bound dehydrogenases, which are linked to the respiratory chain in a manner enabling energy conservation via electron transport phosphorylation (etp) in the cytoplasmic membrane. neither the glucose and gluconic acid nor the xylose and xylonic acid are metabolized. therefore, measurements of sugar ... | 1997 | 9143455 |
| a biosensor for environmental genotoxin screening based on an sos lux assay in recombinant escherichia coli cells. | a genetically controlled luminescent bacterial reporter assay, the sos lux test, was developed for rapid detection of environmental genotoxins. the bioassay is based on the recombinant plasmid ppls-1, which was constructed as a derivative of pbr322, carrying the promoterless luxcdabfe genes of photobacterium leiognathi downstream of a truncated cda gene from cold with a strong sos promoter. e. coli reca+ strains containing this construction are inducible to high levels of light production in the ... | 1997 | 9361425 |
| identification and characterization of the two-enzyme system catalyzing oxidation of edta in the edta-degrading bacterial strain dsm 9103. | in a gram-negative isolate (dsm 9103) able to grow with edta as the sole source of carbon, nitrogen, and energy, the first two steps of the catabolic pathway for edta were elucidated. they consisted of the sequential oxidative removal of two acetyl groups, resulting in the formation of glyoxylate. an enzyme complex that catalyzes the removal of two acetyl groups was purified and characterized. in the reaction, ethylenediaminetriacetate (ed3a) was formed as an intermediate and n,n'-ethylenediamin ... | 1997 | 9371437 |
| purity of recycled fibre-based materials. | in order to study the purity of recycled fibre-based materials, products containing recycled fibre as well as recycled pulp were examined with regard to their chemical impurities, toxicity and microbiological quality. the study was carried out to clarify both qualitatively and quantitatively the variations in microbiological quality. the levels of several classes of chemical substances were analysed and semi-volatile and volatile substances identified. the toxicity and mutagenicity of virgin fib ... | 1997 | 9373533 |
| detection and speciation of bacteria through pcr using universal major cold-shock protein primer oligomers. | the detection of bacteria using pcr is a well-established diagnostic technique. however, conventional pcr requires the use of dna primer oligomers that are specific to the target organism and, as a consequence, a sample can only be tested for the presence of that specific target. a significant advantage would be to probe a sample for the presence of any bacteria, followed by identification. to achieve this it is necessary to identify a dna sequence common to all bacteria. here we demonstrate tha ... | 1997 | 9439003 |
| toxicity of methyl tertiary butyl ether to daphnia magna and photobacterium phosphoreum. | | 1995 | 8555689 |
| methenyltetrahydrofolate cyclohydrolase is rate limiting for the enzymatic conversion of 10-formyltetrahydrofolate to 5,10-methylenetetrahydrofolate in bifunctional dehydrogenase-cyclohydrolase enzymes. | the kinetic properties of three methylenetetrahydrofolate dehydrogenase-cyclohydrolase (d/c) enzymes (the nadp-dependent bifunctional domain of the human cytoplasmic trifunctional enzyme, the human mitochondrial nad-dependent bifunctional enzyme, and the nad(p)-dependent bifunctional enzyme from photobacterium phosphoreum) were determined in both forward and reverse directions. in the forward direction, the enzymes possess widely different ratios of kcat c/kcat d, but all channel methenylh4folat ... | 1998 | 9454603 |
| rapidly fatal infection due to photobacterium (vibrio) damsela. | | 1997 | 9356821 |
| cloning and expression of a marine bacterial beta-galactoside alpha2,6-sialyltransferase gene from photobacterium damsela jt0160. | sialyltransferase 0160, a bacterial sialyltransferase which catalyzes the incorporation of neuac from cmp-neuac into the galactose residue of the carbohydrate chain at position 6, is produced by photobacterium damsela jt0160. the gene coding for sialyltransferase 0160 (bst) was cloned, sequenced, and expressed in escherichia coli. the sialyltransferase 0160 gene contains an open reading frame of 2,028 base pairs encoding a protein of 675 amino acid residues. the deduced amino acid sequence of si ... | 1998 | 9504414 |
| application of an iterative approach for development of a microbial model predicting the shelf-life of packed fish. | an iterative approach was used to develop a microbial model for shelf-life prediction of cod fillets packed in modified atmospheres. the effect of temperature (0-15 degrees c) and co2 (0-100%) on growth of the specific spoilage organism, photobacterium phosphoreum, was studied in packed cod and in liquid media. p. phosphoreum was a dominant part of the spoilage microflora of packed cod stored at the extremes of the range of conditions studied. the organism is therefore likely to be important for ... | 1997 | 9506282 |
| procedure to screen illicit discharge of toxic substances in septic sludge received at a wastewater treatment plant. | this paper presents an integrative approach, using toxicological and chemical analyses, to screen toxic and illegal substances that could be added to the septic sludge transported by a tanker truck to the wastewater treatment plant of the montreal urban community (muc). microtox, lettuce root elongation, and a bacterial respiration test were used to establish the toxicity range of a normal sludge and the determination of threshold limit criteria. septic sludge samples were spiked with different ... | 1998 | 9515073 |
| a new approach to separate the genus photobacterium from vibrio with rflp patterns by hhai digestion of pcr-amplified 16s rdna. | a new approach to separate members of the genus photobacterium from the genus vibrio with rflp (restriction fragment length polymorphism) patterns by hhai digestion of pcr-amplified 16s rdna was developed in the present study. it was clearly shown that these patterns of the genus photobacterium were unique and distinguishable from vibrio species. this method is very simple and does not need other supporting procedures, such as southern transfer and probe hybridization. it can be applied not only ... | 1998 | 9516546 |
| quorum sensing: potential means of treating gram-negative infections? | | 1998 | 9525356 |
| toxicity testing of refinery wastewater using microtox. | | 1998 | 9528705 |
| mass production of bacterial alpha 2,6-sialyltransferase and enzymatic syntheses of sialyloligosaccharides. | to supply alpha 2,6-sialyltransferase for the large-scale synthesis of sialoside, we investigated culture conditions for the production of sialyltransferase 0160. the addition of galactose and beef extract, and control of the ph of the culture medium were effective on the production of sialyltransferase 0160. the maximal enzyme productivity reached 550 units/l. using a crude extract of photobacterium damsela jt0160 cells as an enzyme source, enzymatic syntheses were performed with mono- and di-s ... | 1998 | 9532777 |
| identification of a regulatory protein required for pressure-responsive gene expression in the deep-sea bacterium photobacterium species strain ss9. | here, we report the characterization of a gene necessary for hydrostatic pressure regulation of gene expression in the deep-sea bacterium photobacterium species strain ss9. the deduced amino acid sequence of the gene product shares extensive similarity to toxr, a transmembrane dna-binding protein first discovered as a virulence determinant in the pathogenic bacterium vibrio cholerae. changes in hydrostatic pressure induce changes in both the abundance and the activity of the ss9 toxr protein (or ... | 1998 | 9535087 |
| characteristics analysis of the luza gene encoding chaperone from photobacterium leiognathi related to bioluminescence. | nucleotide sequence of the luza gene (genbank accession no. af039303) from photobacterium leiognathi atcc 25521 (ncimb 2193) has been determined, and the chaperone encoded by the luza gene was deduced. the luza chaperone has a calculated m(r) 26,295 and comprises 230 amino acid residues; the hydrophobic alpha-helix n-terminal 21 amino acid residues mkktifallfmsvfi sypsfa is the leader peptide, therefore the matured luza chaperone has a calculated m(r) 23,871 and comprises 209 amino acid residues ... | 1998 | 9535753 |
| evaluation of the toxicity of guarana with in vitro bioassays. | a natural stimulant, paullinia cupana, commonly called guarana, was tested for its ability to induce in vitro toxicity in chinese hamster ovary (cho) cells and bacterial cells (photobacterium phosphoreum). the cytotoxic effects of aqueous guarana extracts were evaluated by three endpoint systems: neutral red (nr) uptake assay, total protein content [kenacid blue (kb)] assay, and tetrazolium (mtt) assay. the microtox test was also used. results indicated that the lowest concentration of guarana t ... | 1998 | 9570905 |
| characteristic analysis of the luxg gene encoding the probable flavin reductase that resides in the lux operon of photobacterium leiognathi. | nucleotide sequence of the luxg gene (genbank accession no. af053227) from photobacterium leiognathi pl741 has been determined, and the encoded probable flavin reductase is deduced. the probable flavin reductase encoded by the luxg gene has a calculated m(r) 26,544 and comprises 235 amino acid residues. the probable flavin reductase like the nad(p)h-flavin reductase might catalyze the reduction of flavins. alignment and comparison of the probable flavin reductases from p. leiognathi pl741, atcc ... | 1998 | 9610381 |
| toxicity and mutagenicity of component classes of oils isolated from soils at petroleum- and creosote-contaminated sites. | microtox and ames bioassays were employed to assess acute toxicity and mutagenicity of water soluble components of class-fractionated oils extracted from one creosote- and four petroleum-contaminated soils. microtox results revealed that potential acute toxicity resides mainly in the polar class fractions at three sites and indicated potential synergistic and antagonistic effects between compounds in the total extracts at two sites. ames salmonella/microsome testing indicated that the polyaromat ... | 1997 | 9448516 |
| the efficiency of different phenol-degrading bacteria and activated sludges in detoxification of phenolic leachates. | phenolic composition, toxicity and biodegradability of three different phenolic leachates/samples was studied. samples a and c were the leachates from the oil-shale industry spent shale dumps at kohtla-järve, estonia. sample b was a laboratory-prepared synthetic mixture of 7 phenolic compounds mimmicking the phenolic composition of the leachate a. toxicity of these 3 samples was analyzed using two photobacterial test (biotox and microtox), daphnia test (daphtoxkit f pulex) and rotifiers' test (r ... | 1998 | 9650267 |
| the luxr regulator protein controls synthesis of polyhydroxybutyrate in vibrio harveyi. | the luxr regulatory protein of vibrio harveyi has been shown to control synthesis of polyhydroxybutyrate (phb) as well as luminescence so as to occur at high cell density, suggesting that it is a general regulatory protein. mutants defective in the production of luxr (d1, d34, and mr1130) were found to be missing phb, whose synthesis could be restored by complementation with luxr. triparental mating with a v. harveyi genomic library revealed the presence of three genomic clones (g1, g2 and g3) t ... | 1998 | 9659397 |
| the detection and characterization of genetically programmed responses to environmental stress. | the rapid, accurate, and inexpensive detection of environmental contaminants (and the stress that they engender) is still a major problem worldwide. though assays exist for monitoring these pollutants, they can often be expensive, time-consuming, and require extensive equipment and/or training in order to be effective. research over the past decade has pointed to the measurement of enzymes encoded by genes programmed to respond to particular classes of environmental stress as a means of quantify ... | 1998 | 9668622 |
| photobacterium profundum sp. nov., a new, moderately barophilic bacterial species isolated from a deep-sea sediment. | a novel, moderately barophilic bacterium was isolated from a sediment sample obtained from the ryukyu trench, at a depth of 5110 m. the isolate, designated strain dsj4, is a gram-negative rod capable of growth between 4 degrees c and 18 degrees c under atmospheric pressure, with optimum growth displayed at 10 degrees c, and capable of growth at pressures between 0.1 mpa and 70 mpa at 10 degrees c, with optimum growth displayed at 10 mpa. strain dsj4 is a moderately barophilic bacterium, and show ... | 1998 | 9676237 |
| the extracytoplasmic function sigma factors: role and regulation. | alternative sigma factors provide a means of regulating gene expression in response to various extracellular changes. one such class of sigma factors appears to control a variety of functions, including expression of heat-shock genes in escherichia coli, biosynthesis of alginates and carotenoids in pseudomonas aeruginosa and myxococcus xanthus, respectively, iron uptake in e. coli and pseudomonas spp., nickel and cobalt efflux in alcaligenes europhus, plant pathogenicity in pseudomonas syringae ... | 1998 | 9680198 |
| the molecular biology of barophilic bacteria. | many microorganisms from the deep-sea display high-pressure-adapted--also described as barophilic or piezophilic--growth characteristics. phylogenetic studies have revealed that a large proportion of the barophilic bacteria currently in culture collections belong to a distinct subgroup of the genus shewanella, referred to as the "barophile branch." many of the basic properties of barophiles that enable their survival at extremes of pressure remain to be elucidated. however, several genes whose e ... | 1997 | 9680316 |
| toxicity assessment of pesticides using the microtox test: application to environmental samples. | | 1997 | 9307428 |
| [the evaluation of the total toxicity of heavy metals based on the luminescence bacterial test]. | | 1998 | 9721511 |
| cu,zn superoxide dismutase from photobacterium leiognathi is an hyperefficient enzyme. | the catalytic rate constant of recombinant photobacterium leiognathi cu,zn superoxide dismutase has been determined as a function of ph by pulse radiolysis. at ph 7 and low ionic strength (i = 0.02 m) the catalytic rate constant is 8.5 x 10(9) m-1 s-1, more than two times the value found for all the native eukaryotic cu,zn superoxide dismutases investigated to date. similarly, brownian dynamics simulations indicate an enzyme-substrate association rate more than two times higher than that found f ... | 1998 | 9724543 |
| on the coordination and oxidation states of the active-site copper ion in prokaryotic cu,zn superoxide dismutases. | the active-site copper ion of the prokaryotic cu,zn superoxide dismutase from p. leiognathi is found to undergo reversible reduction upon irradiation of the protein solution with a high-intensity x-ray beam from a third-generation synchrotron source. the same phenomenon is observed for the enzyme crystals, whose diffraction pattern has been obtained from synchrotron sources. in this case the active-site copper-ligand coordination bond lengths and in particular the cu-ne2(his61) distance are cons ... | 1998 | 9731178 |
| the effect of suspending solution supplemented with marine cations on the oxidation of biolog gn microplate substrates by vibrionaceae bacteria. | bacteria belonging to the family vibrionaceae were suspended using saline and a solution prepared from a marine-cations supplement. the effect of this on the profile of oxidized substrates obtained when using biolog gn microplates was investigated. thirty-nine species belonging to the genera aeromonas, listonella, photobacterium, and vibrio were studied. of the strains studied, species of listonella, photobacterium, and vibrio could be expected to benefit from a marine-cations supplement that co ... | 1998 | 9643966 |
| thawed cod fillets spoil less rapidly than unfrozen fillets when stored under modified atmosphere at 2 degrees c. | the effect of two months of frozen storage at -20 degrees c on the spoilage characteristics and shelf life of thawed and modified atmosphere packed (map) cod fillets stored at 2 degrees c was studied. thawed map cod fillets were compared with fresh cod fillets stored in co2-containing modified atmospheres with and without added oxygen. the shelf life of 11 to 12 days in the fresh map cod was extended to more than 20 days in the thawed map cod at 2 degrees c. this shelf life extension was most li ... | 1998 | 9766063 |
| specific inhibition of photobacterium phosphoreum extends the shelf life of modified-atmosphere-packed cod fillets. | inhibition of the specific spoilage organism, photobacterium phosphoreum, was studied in model substrates and in modified-atmosphere-packed cod fillets. the objective was to determine how inhibition of this organism influenced spoilage. the spoilage reactions limiting shelf life were studied rather than the development of a new product. in naturally contaminated modified-atmosphere-packed cod fillets, 500 ppm na2caedta reduced the growth rate of p. phosphoreum by 40% and shelf life was increased ... | 1998 | 9766075 |
| established technologies and new approaches in applying luminous bacteria for analytical purposes. | | 1997 | 9481605 |
| microtox system, a new approach to the safety evaluation of medical devices. | medical devices of both intra- and extra-corporeal applications may contain leachable toxicants that warrant routine testing using internationally recognized/practiced protocols. an in vitro microtox system comprising a selected bioluminescence strain of bacteria-photobacterium phosphorium which emits light as an end product of its respiration, is used to screen out toxicants from biomaterials/devices. a microtox system was standardized and validated in our laboratory conditions. our experiences ... | 1998 | 9777466 |
| mechanism of reduced flavin transfer from vibrio harveyi nadph-fmn oxidoreductase to luciferase. | the mechanisms of reduced flavin transfer in biological systems are poorly understood at the present. the vibrio harveyi nadph-fmn oxidoreductase (frp) and the luciferase pair were chosen as a model for the delineation of the reduced flavin transfer mechanism. frp, which uses fmn as a cofactor to mediate the reduction of the flavin substrate by nadph, exhibited a ping-pong kinetic pattern with a km, fmn of 8 microm and a km,nadph of 20 microm in a single-enzyme spectrophotometric assay monitorin ... | 1998 | 9772191 |
| use of an autobioluminescent salmonella hadar to monitor the effects of acid and temperature treatments on cell survival and viability on lactic acid-treated poultry carcasses. | to determine the long-term effects of a lactic acid rinse on viability and recovery of pathogens, salmonella hadar was isolated from poultry and bioluminescent constructs obtained by transformation with the lux (cdabe) gene cassette from photobacterium phosphoreum. results indicated that the transformed salmonella hadar lux was otherwise phenotypically similar to the wild-type strain. viability studies were performed by measuring luminescence following lactic acid treatment of turkey breast and ... | 1998 | 9829182 |
| survival of fish-virulent strains of photobacterium damselae subsp. damselae in seawater under starvation conditions. | the survival of fish-virulent strains of photobacterium damselae subsp. damselae in seawater microcosms, with and without sediment, was investigated. the strains survived as culturable bacteria at 14 and 22 degrees c for at least 1 year, and infectivity for fish was maintained. at 5 degrees c, cells lost culturability on solid media, but this was recovered when the temperature was increased to 22 degrees c. finally, morphological changes in the bacterium (rod to coccus), and production of vesicl ... | 1998 | 9835027 |
| differentiation of marine luminous bacteria using commercial identification plates. | differentiation of marine luminous bacteria using biology gn plate combined with api 20e or the bbl crystal id plate inoculated with cell suspensions in artificial seawater was accomplished by comparison to type species using cluster analysis. inoculum density affected the results from biolog gn plates, but had less of an effect on the reactions obtained from api 20e strip or bbl crystal id plate. in a few cases, combination of the biolog gn traits along with either the api 20e or crystal id tra ... | 1998 | 9693982 |
| ecotoxicological assessment of ashes and particulate matter from fluidized bed combustion of coal. | toxicity of ash and particulate matter from the gases generated in coal fluidized bed combustion (fbc) has been determined by the microtox assay according to the standard leachate procedure. results are compared with the polycylic aromatic hydrocarbon (pah) content of the particulate matter, which was determined by fluorescence spectroscopy. although pahs are considered highly toxic compounds, the low ecotoxicity values obtained could be explained by the low solubility of the compounds in water. ... | 1998 | 9756690 |
| evolutionary constraints for dimer formation in prokaryotic cu,zn superoxide dismutase. | prokaryotic cu,zn superoxide dismutases are characterized by a distinct quaternary structure, as compared to that of the homologous eukaryotic enzymes. here we report a newly determined crystal structure of the dimeric cu,zn superoxide dismutase from photobacterium leiognathi (crystallized in space group r32, refined at 2.5 a resolution, r-factor 0.19) and analyse it in comparison with that of the monomeric enzyme from escherichia coli. the dimeric assembly, observed also in a previously studied ... | 1999 | 9878406 |
| the anti-sigma factors. | a mechanism for regulating gene expression at the level of transcription utilizes an antagonist of the sigma transcription factor known as the anti-sigma (anti-sigma) factor. the cytoplasmic class of anti-sigma factors has been well characterized. the class includes asia form bacteriophage t4, which inhibits escherichia coli sigma 70; flgm, present in both gram-positive and gram-negative bacteria, which inhibits the flagella sigma factor sigma 28; spoiiab, which inhibits the sporulation-specific ... | 1998 | 9891799 |
| reassessment of the taxonomic position of vibrio iliopiscarius (onarheim et al. 1994) and proposal for photobacterium iliopiscarium comb. nov. | the phylogenetic position of vibrio iliopiscarius was inferred by the maximum-likelihood, maximum-parsimony and neighbour-joining methods on the basis of almost complete 16s rrna gene sequences. the results showed that this species falls into the same cluster as photobacterium species and is clearly distinct from other vibrio species. its nearest phylogenetic neighbour is photobacterium phosphoreum. from these results, it is concluded that v. iliopiscarius should be reclassified as photobacteriu ... | 1999 | 10028270 |
| superoxide dismutase and catalase in photobacterium damselae subsp. piscicida and their roles in resistance to reactive oxygen species. | photobacterium damselae subsp. piscicida (formerly pasteurella piscicida) is the causative agent of pasteurellosis or pseudotuberculosis in warm water marine fish. enzymes which neutralize reactive oxygen species, produced during aerobic metabolism or during respiratory burst in fish macrophages, are important virulence factors in many pathogens. this study characterizes a periplasmic superoxide dismutase (sod) and a cytoplasmic catalase in p. damselae. purification and partial amino-terminal se ... | 1999 | 10075430 |
| toward the engineering of a super efficient enzyme. | the catalytic activity of a mutant of photobacterium leiognathi cu, zn superoxide dismutase in which the glu59 residue, conserved in most bacterial variants of the enzyme, has been replaced by glutamine was investigated by pulse radiolysis. at neutral ph the enzyme was found to have a kcat/km of 1.0 +/- 0.1 x 10(10) m-1s-1 the highest value ever found for any superoxide dismutase. brownian dynamics simulation suggests that such a high value is due to an enhanced substrate attraction by the modif ... | 1999 | 10079201 |
| monounsaturated but not polyunsaturated fatty acids are required for growth of the deep-sea bacterium photobacterium profundum ss9 at high pressure and low temperature. | there is considerable evidence correlating the production of increased proportions of membrane unsaturated fatty acids (ufas) with bacterial growth at low temperatures or high pressures. in order to assess the importance of ufas to microbial growth under these conditions, the effects of conditions altering ufa levels in the psychrotolerant piezophilic deep-sea bacterium photobacterium profundum ss9 were investigated. the fatty acids produced by p. profundum ss9 grown at various temperatures and ... | 1999 | 10103272 |
| recd function is required for high-pressure growth of a deep-sea bacterium. | a genomic library derived from the deep-sea bacterium photobacterium profundum ss9 was conjugally delivered into a previously isolated pressure-sensitive ss9 mutant, designated ec1002 (e. chi and d. h. bartlett, j. bacteriol. 175:7533-7540, 1993), and exconjugants were screened for the ability to grow at 280-atm hydrostatic pressure. several clones were identified that had restored high-pressure growth. the complementing dna was localized and in all cases found to possess strong homology to recd ... | 1999 | 10197992 |
| polyphasic approach to the characterisation of marine luminous bacteria. | fifteen luminous bacterial strains were isolated from the tyrrhenian sea coastal waters off northeastern sicily and characterised by a combination of phenotypic and molecular tests in order to identify them and to determine their intraspecific genetic variability. five luminous type strains, vibrio splendidus ncimb 1, v. harveyi ncimb 1280, v. fischeri ncimb 1281, v. orientalis ncimb 2195 and photobacterium leiognathi ncimb 2193, were used as reference. on the basis of their phenotypic character ... | 1999 | 10229952 |
| characterization of the spectroscopic properties of the cu,co cluster in a prokaryotic superoxide dismutase. | a cu,co derivative of the cu,znsod from photobacterium leiognathi, in which cobalt has been selectively substituted for zinc, has been prepared and spectroscopically investigated. the derivative shows three bands in the visible region at 530, 566, and 600 nm when copper is in the oxidized state. reduction or depletion of the copper ion produce a shift of the band absorbing at 600 to 590 nm because of the detachment from copper of the imidazolate bridging the two metals when copper is in the oxid ... | 1999 | 10334865 |
| vibrios associated with litopenaeus vannamei larvae, postlarvae, broodstock, and hatchery probionts. | several bacteriological surveys were performed from 1994 to 1996 at different litopenaeus vannamei hatcheries (in ecuador) and shrimp farms (in mexico). samples were taken from routine productions of healthy and diseased l. vannamei larvae, postlarvae, and their culture environment and from healthy and diseased juveniles and broodstock. in ecuador, the dominant bacterial flora associated with shrimp larvae showing symptoms of zoea 2 syndrome, mysis mold syndrome, and bolitas syndrome has been de ... | 1999 | 10347048 |
| [lipopolysaccharide from photosynthetic bacteria blocks ca2+ response of human neutrophils induced by endotoxins from shigella flexneri and its re-mutant]. | | 1999 | 10347828 |
| applicability of catalyst in ecotoxicology, a new promising tool for 3d-qsar: study of chlorophenols. | with the aim of applying recent quantitative structure-activity relationship (qsar) descriptors coming from the pharmaceutical sciences to the modelization of ecotoxicity data, this work deals with the establishment of 3d-qsar on chlorophenols by using for the first time in this field the recent catalyst software. the training set has been intentionally chosen simply because of its homogeneous character and the abundance of reliable experimental values. among 69 data sets collected in the litera ... | 1999 | 10381301 |
| purification and characterization of nfra1, a bacillus subtilis nitro/flavin reductase capable of interacting with the bacterial luciferase. | ipa-43d is a hypothetical gene identified by the bacillus subtilis genome project (mol. microbiol. 10, 371-384 1993; nature 390, 249-256 1997). the ipa-43d protein overexpressed in e. coli was purified to homogeneity and its properties were analyzed biochemically. the ipa-43d protein was found to be tightly associated with fmn and to be capable of reducing both nitrofurazone and fmn effectively. although the ipa-43d protein catalysis obeys the ping-pong bi-bi mechanism, catalysis mode was change ... | 1998 | 9836433 |
| viable heterotrophic bacteria in water and sediment in 'mar piccolo' of taranto (ionian sea, italy). | samples of water and sediment were collected from october 1996 to september 1997 in 'mar piccolo' of taranto (ionian sea, italy). mar piccolo is a semi-enclosed basin subject to pollution and receives a considerable amount of sewage and industrial waste. qualitative and quantitative analyses of the composition of the microbial flora were conducted on samples from six stations. the highest bacterial densities, in water and in sediment samples, were found in summer and the lowest, in autumn. among ... | 1999 | 10389241 |
| prediction of water solubility and toxicity of substituted indoles to photobacterium phosphoreum using molecular connectivity indices and quantum chemical parameters. | | 1998 | 9841718 |
| occurrence of p-flavin binding protein in vibrio fischeri and properties of the protein. | in previous studies involving photobacterium species we proposed that (i) p-flavin is the product of luciferase, (ii) the physiological function of the lux operon is not to produce light but to produce fp(390) (luxf protein), including its prosthetic group, p-flavin, and (iii) fp(390) reactivates oxidatively inactivated cobalamin-dependent methionine synthase similar to flavodoxin but at relatively high ionic strength. it seems difficult to extend this idea to all luminous bacteria because the l ... | 1999 | 10423522 |
| luminous bacteria cultured from fish guts in the gulf of oman. | the incidence of culturable luminous bacteria in omani market fish guts was correlated to habitat type amongst 109 species of fish. isolated representative luminous bacteria were compared to known species using the biolog system (95 traits/isolate) and cluster analysis, which showed that the main taxa present in fish guts were clades related to vibrio harveyi and photobacterium species with sporadic incidence of p. phosphoreum. the luminous isolates from gut of the slip-mouth (barred pony fish), ... | 1999 | 10423577 |
| phenotypic characterization of the marine pathogen photobacterium damselae subsp. piscicida. | the taxonomic position of photobacterium damselae subsp. piscicida, the causative agent of fish pasteurellosis, is controversial as this organism has also been described as 'pasteurella piscicida'. to clarify the taxonomic position of the pathogen, a total of 113 p. damselae subsp. piscicida strains and 20 p. damselae subsp. damselae strains, isolated from different geographical areas and from the main affected fish species, were analysed using 129 morphological and biochemical tests, including ... | 1998 | 9828416 |
| evidence of stable monomeric species in the unfolding of cu,zn superoxide dismutase from photobacterium leiognathi. | the equilibrium unfolding process of photobacterium leiognathi cu,zn superoxide dismutase has been quantitatively monitored through circular dichroism (cd) and fluorescence spectroscopy, upon increasing the guanidinium hydrochloride concentration. the study has been undertaken for both the holo- and the copper-free derivative to work out the role of copper in protein stability. in both cases the unfolding was reversible. the denaturation curve derived from cd and fluorescence spectroscopy was no ... | 1999 | 10510278 |
| autoinduction of light emission in different species of bioluminescent bacteria. | the production of light in most luminescent bacteria lags behind cellular growth with induction of luminescence only occurring at high cell density. the maximum intensity and the cell density reached before induction of luminescence is highly dependent on the bacterial species and the growth conditions including temperature, salt and nutrient composition as well as other experimental conditions. although common n-acylhomoserine lactone autoinducers have been identified in vibrio (photobacterium) ... | 1999 | 10398554 |
| 16s rrna gene sequence analysis of photobacterium damselae and nested pcr method for rapid detection of the causative agent of fish pasteurellosis. | the causative agent of fish pasteurellosis, the organism formerly known as pasteurella piscicida, has been reclassified as photobacterium damselae subsp. piscicida on the basis of 16s rrna gene sequence comparisons and chromosomal dna-dna hybridization data; thus, this organism belongs to the same species as photobacterium damselae subsp. damselae (formerly vibrio damselae). since reassignment of p. damselae subsp. piscicida was based on only two strains, one objective of the present work was to ... | 1999 | 10388687 |
| transfer of xenobiotics through cell membranes of luminous bacteria. | the influence of some chemical substances on luminous bacteria was studied to elucidate the interrelation between the xenobiotics action on bacterial luminescence and cell ultrastructure. such substances as quinones, phenols, chlorides of heavy metals (in concentrations of substances inhibiting luminescence by 50%) resulted in damaging effects upon bacteria: a lot of cells had damage of membranes due to changes in their permeability. it was found that the high concentration of edta and toluene d ... | 1999 | 10512991 |
| assessment of a magnetic bead-eia based kit for rapid diagnosis of fish pasteurellosis. | the accuracy of the magnetic beads-eia based bionor aquaeia-pp kit for the rapid diagnosis of pasteurellosis was evaluated. the kit reacted with all the photobacterium damselae subsp. piscicida strains included in this study, with a detection limit of 10(4) bacteria/ml. however, non-specific reactions were observed with isolates of ph. damselae subsp. damselae or ph. histaminum when the bacterial concentration was high (10(9)-10(10) bacteria/ml). similar findings in specificity and sensitivity w ... | 1999 | 10520595 |
| bioluminescent method to determine non-specific endotoxicosis in therapy. | the possibility of using a bioluminescence method for measuring endotoxicosis during therapy has been investigated. results of our experiments shown that a bioluminescence method can be used as a reliable criterion to monitor the course of disease for patients with bronchitis, ulcerous disease or chronic colecystitis. assays using both soluble and immobilized reagents are possible. this method does not reveal differences in patients with sepsis, hepatocirrhosis or oncology. the methods are highl ... | 1999 | 10441049 |
| a study of deep-sea natural microbial populations and barophilic pure cultures using a high-pressure chemostat. | continuous cultures in which a high-pressure chemostat was used were employed to study the growth responses of (i) deep-sea microbial populations with the naturally occurring carbon available in seawater and with limiting concentrations of supplemental organic substrates and (ii) pure cultures of copiotrophic barophilic and barotolerant deep-sea isolates in the presence of limiting carbon concentrations at various pressures, dilution rates, and temperatures. we found that the growth rates of nat ... | 1999 | 10583982 |
| acetone formation in the vibrio family: a new pathway for bacterial leucine catabolism. | there is current interest in biological sources of acetone, a volatile organic compound that impacts atmospheric chemistry. here, we determined that leucine-dependent acetone formation is widespread in the vibrionaceae. sixteen vibrio isolates, two listonella species, and two photobacterium angustum isolates produced acetone in the presence of l-leucine. shewanella isolates produced much less acetone. growth of vibrio splendidus and p. angustum in a fermentor with controlled aeration revealed th ... | 1999 | 10601206 |
| [structural approach to simulating the population dynamics of unstable recombinant strains of bacteria containing multicopy plasmids]. | | 1999 | 10613258 |
| a region of the transmembrane regulatory protein toxr that tethers the transcriptional activation domain to the cytoplasmic membrane displays wide divergence among vibrio species. | the virulence regulatory protein toxr of vibrio cholerae is unique in that it contains a cytoplasmic dna-binding-transcriptional activation domain, a transmembrane domain, and a periplasmic domain. although toxr and other transmembrane transcriptional activators have been discovered in other bacteria, little is known about their mechanism of activation. utilizing degenerate oligonucleotides and pcr, we have amplified internal toxr gene sequences from seven vibrio and photobacterium species and s ... | 2000 | 10629204 |