Genetic Diversity of Algal Viruses Which Lyse the Photosynthetic Picoflagellate Micromonas pusilla (Prasinophyceae)

The genetic similarity among eight clones of Micromonas pusilla virus (MpV) isolated from five geographic locations was measured by DNA hybridization. Our objective was to explore the existence of genetically distinct populations of MpV by comparing the similarity among MpVs isolated from a single water sample to the similarity among viruses isolated from geographically distant locations. The highest and lowest similarities we observed were 70% (plusmn) 1.1% (mean (plusmn) standard error [SE], n = 3) for virus strains SP1 and SP2 isolated from a California coastal water sample and 13% (plusmn) 1.9% for strains SP2 and PB6; the latter was isolated from New York estuarine water. However, the similarity between MpV isolated from a single water sample was not always greater than the similarity between viruses isolated from different locations. Viruses PB7 and PB8 were isolated from a single New York estuarine sample but were only 16% (plusmn) 0.5% similar, whereas PB7 was quite similar (43% (plusmn) 2.9%) to PL1, a virus from Texas coastal water. Overall, the similarity among MpVs isolated from a single geographic location, 34% (plusmn) 12.6% (mean (plusmn) SE, n = 4), was not significantly different from the similarity among MpVs isolated from geographically distant locations, 26.6% (plusmn) 2.7% (mean (plusmn) SE, n = 24) (P = 0.92, Mann-Whitney U test). Clones of MpV were more similar to each other than they were to the related algal virus PBCV-1, and three groups of MpVs consisting of (i) PL1, SG1, PB6, and PB7, (ii) PB8, and (iii) GM1, SP1, and SP2 were resolved. The genetic variation among MpVs isolated from a single water sample was as large as the variation between viruses isolated from different oceans. If MpVs within a geographic location share genetic characteristics not shared with MpVs from geographically distant locations, this was not reflected in the overall similarity of their genomes.     

Differentiation of methicillin-resistant strains of Staphylococcus aureus by prophage specificity]

By inducing with mitomycin C the following phages were isolated from all the tested 32 methicillin resistant strains of S. aureus: the serogroup B phage was isolated from 2 strains, the serogroup B and F phages were isolated from 5 strains and the serogroup F phage was isolated from 25 strains. The phages were divided into 5 groups by the antiphage immunity. In group 1 of the phages 4 additional phages were specified. By the specificity of the prophages in the cultures all the strains were divided into 5 groups. Group 1 of the cultures was divided into 5 subgroups (A, B, C, D and E).     

Kaempferol glycosides from Siparuna apiosyce

The kaempferol derivative 3,7-di-O-methyl-4'-O-beta-[alpha rhamnosyl (1 --> 6)]-glucopyranoside (siparunoside) was isolated from the leaves of Sparuna apiosyce. Its structure was established by extensive NMR studies. The alkaloids reticuline and liriodenine were also isolated from the leaves along with the kaempferol derivative tiliroside. Benzylisoquinoline alkaloids were isolated from the wood (liriodenine) and wood bark (liriodenine, laurotetanine, N-methyl-laurotetanine, reticuline), together with a mixture of cis and trans-N-feruloyltyramines. 3,7,4'-tri-O-methylkaempferol was isolated from all organs.     

海洋放线菌Streptomyces variabilis strain 6-1次级代谢产物的研究

目的:对来自海洋软珊瑚的链霉菌6-1(Streptomyces variabilis strain 6-1)进行次级代谢产物的分离和鉴定,寻找具有生物活性的化合物,为人类健康服务。方法:采用液体培养基对分自海洋软珊瑚Scleronephthya sp中的链霉菌6-1(Streptomyces variabi-lis strain 6-1)进行发酵培养,用乙酸乙酯对发酵液进行萃取;采用半制备高效液相色谱(semi-preparative HPLC)分离方法对乙酸乙酯萃取物进行分离纯化,得到单体化合物;运用电喷雾质谱(ESI-MS)、核磁共氢振(1H NMR)、核磁共振碳谱(13C NMR)和物理性质对所得单体化合物进行结构鉴定。结果:从海洋链霉菌6-1(strain 6-1)发酵液的乙酸乙酯萃取物中分离得到3个单体化合物,分别鉴定为:7,4'-二羟基异黄酮(1)、5,7,4'-三羟基异黄酮(2)和丁烯酸内酯-Ⅰ(3)。结论:丁烯酸内酯-Ⅰ是从链霉菌属首次分离得到,化合物1和2均是从Streptomyces variabilis中首次分离得到;变异链霉菌6-1(Streptomyces variabilis strain 6-1)可以作为活性化合物3(丁烯酸内酯-Ⅰ)的重要来源。     

Alterations in the longevity and fecundity of adult Trichinella pseudospiralis related to method of isolation of infective larvae

The infectivity of Trichinella pseudospiralis infective larvae was reduced significantly following exposure to low pH or a combination of 1% pepsin at low pH compared to that for larvae isolated in phosphate-buffered saline (PBS) at pH 7.0. Reduction of host gastric pH by administration to mice of sodium bicarbonate solution in PBS was accompanied by an increase in the infectivity of larvae isolated in 1% pepsin/HCl (P/HCl) compared to that for worms inoculated into hosts given PBS alone. Fewer adult worms developing from larvae isolated in P/HCl became established in the host small bowel than was seen with larvae isolated in PBS; moreover, the fecundity in vitro of adult worms developing from P/HCl-isolated larvae was reduced below that for adults developing from larvae isolated from host muscle in PBS. More adult worms were recovered following infection of immune hosts with PBS-isolated larvae than were recovered from immune mice challenged with larvae isolated in P/HCl. Similar findings were observed in mice immunized by infection with Trichinella spiralis and challenged with T. pseudospiralis larvae isolated in either P/HCl or PBS. Immunization of mice with T. pseudospiralis larvae isolated by either method and challenged with larvae of T. spiralis resulted in recovery of similar percentages of the challenge inoculum.     

Isolation of a human X chromosome-linked gene essential for progression from G1 to S phase of the cell cycle

The tsBN462 cell line, a temperature-sensitive (ts) mutant isolated from the hamster cell line, BHK21/13, cannot progress into S phase at 39.5 degrees C, following the release from isoleucine deprivation. The mutant cells were transfected with high molecular weight (HMW) DNA from human KB cells, and several human DNA bands were found to be conserved through three cycles of ts+ transformation. Conserved human DNA was isolated from the cosmid library of the secondary ts+ transformant (K-1-1), using 32P-labelled total human DNA as a probe. The isolated human DNA covers about 70 kb of human DNA flanked with hamster DNA, and originates from the human X chromosome. The middle part (56 kb) of the isolated human DNA was conserved through the primary, secondary and tertiary ts+ transformation, without gross rearrangement.     

Chemical and biological characterisation of sapinopyridione, a phytotoxic 3,3,6-trisubstituted-2,4-pyridione produced by Sphaeropsis sapinea, a toxigenic pathogen of native and exotic conifers, and its derivatives

A phytotoxic trisubstituted 2,4-pyridione, named sapinopyridione, was isolated from the culture filtrates of Sphaeropsis sapinea, a fungal pathogen of conifers occurring world-wide. Three strains were isolated from two cypress species. Strain D-55 isolated from Cupressus sempervirens resulted high producer of sapinopyridione (12.3 mg l(-1)), whereas strain D-54 isolated from the same cypress species was low producer (1.1 mg l(-1)); strain D-50 isolated from C. macrocarpa was intermediate producer (5.4 mg l(-1)). Sapinopyridione was characterised by spectroscopic and chemical methods, as the 6-methyl-2-(2-methyl-1-oxobutyl)-1-oxa-5-azaspiro[2.5]oct-6-ene-4,8-dione. The structure was supported by the preparation of three key derivatives, whose phytotoxic and antimycotic activities were also tested on host plants and on three Seiridium species, virulent fungal agents of cypress canker disease. Some structure-activity relationships were identified for both phytoxicity and antifungal activities. These activities appear related to the presence of both pyridione and oxiran rings. Also the carbonyl group of the side chain seems to play a role into impart activity.     

Detection of Shiga toxins,intimin and enterohemolysin in Escherichia coli strains isolated from children in eastern Slovakia

FiftyEscherichia coli strains isolated from stool samples of 51 healthy children, 143 strains isolated from stool samples of 327 children with diarrhea and 24 strains isolated from stool samples of 21 children with suspected hemolytic uremic syndrome were examined for the presence of Shiga toxin-producingE. coli virulence factors (shiga toxin 1 and 2, intimin and enterohemolysin) and their genes. Vero-cell assay and latex agglutination were used for detection of Shiga toxin 1 and 2, TSB agar with washed erythrocytes was used for detection of enterohemolysin; genes encoding shiga toxin 1 and 2, intimin and enterohemolysin were detected using multiplex PCR. The presence ofE. coli strains harboring genes encoding shiga toxin 1 and 2 (12 strains), intimin (34 strains) and enterohemolysin (12 strains) was demonstrated.     

Formation of mercapturic acids in rats after the administration of aralkyl esters

1. Benzylmercapturic acid and hippuric acid were isolated from the urine of rats that had been injected subcutaneously with benzyl acetate. 2. 1-Menaphthylmercapturic acid and 1-naphthoic acid were isolated from the urine of rats after the subcutaneous injection of each of the following compounds: 1-menaphthyl alcohol and its acetate, propionate, butyrate and benzoate esters. 3. A quantitative method for determining 1-menaphthylmercapturic acid in urine was developed and used to measure the excretion of this compound in the urine of rats in the 4-day period after the subcutaneous injection of 1-menaphthyl alcohol and its acetate, propionate, butyrate and benzoate esters. 4. Chromatographic evidence was obtained for the presence of S-(1-menaphthyl)glutathione and S-(1-menaphthyl)-l-cysteine in bile collected from rats with cannulated bile ducts after the animals had been injected subcutaneously with each of the following compounds: S-(1-menaphthyl)glutathione, 1-menaphthyl acetate, propionate and butyrate. 5. Benzylmercapturic acid and 1-menaphthylmercapturic acid were isolated from the urine of rats that had been injected with sodium benzyl sulphate and sodium 1-menaphthyl sulphate respectively.     

Structural and immunological characterization of type IV collagen isolated from chicken tissues

Previously, a type IV collagen fraction was isolated from chicken gizzard and further fractionated into three components called F1, F2 and F3 [Mayne, R. and Zettergren, J.G. (1980) Biochemistry, 19, 4065-4072]. F1 and F2 were consistently isolated in a 2:1 proportion, and the existence of a small native fragment of structure (F1)2F2 was proposed. In the present series of experiments, a type IV collagen fraction was isolated from the chicken kidney and shown to consist almost entirely of F1 and F2 which were again present in a 2:1 proportion. Identical one-dimensional peptide maps for F1 and F2 from both sources were obtained by polyacrylamide gel electrophoresis of peptides obtained after cleavage with cyanogen bromide or Staphylococcus aureus V8 protease. The denaturation temperature of a preparation containing F1 and F2 in native form was determined by optical rotatory dispersion and a single melting curve was observed with a melting temperature of 33 degrees C. This result provides further supportive evidence that F1 and F2 exist as a native fragment (F1)2F2. Antibodies were prepared in rabbits against a type IV collagen fraction isolated from chicken gizzard, and immunofluorescent staining of a wide variety of basement membranes was demonstrated. Experiments were performed in which various type IV collagen fractions were observed in the electron microscope after rotary shadowing. The lengths of (F1)2F2 and F3 were 147 nm and 174 nm respectively, the sum of these lengths (321 nm) corresponding closely to the length of the major triple-helical domain of type IV collagen (326-328 nm). A specific cleavage site was located at a distance of 215 nm from the 7-S domain which, together with the length of (F1)2F2, gives a total length of 362 nm. This value corresponds closely to the maximum length of the arms which originate from the 7-S domain (355 nm) when type IV collagen was solubilized with a low concentration of pepsin. The results suggest that (a) type IV collagen isolated from the chicken gizzard is closely related, if not identical, to type IV collagen isolated from other tissues; (b) there is a single type IV collagen molecule of chain organization[alpha 1(IV)]2 alpha2(IV); (c) the order of the pepsin-resistant fragments within a type IV molecule is 7S-F3-(F1)2F2.     

Isolation and identification of dihydrochrysanolide and its 1-epimer from Chrysanthemum coronarium L

A new sesquiterpene lactone (1) was isolated with known dihydrochrysanolide derivatives (2 and 3) from the flowers of Chrysanthemum coronarium L., and their structures were identified by spectroscopic data. The stereochemistry of the epimers (1 and 2) was determined from NOESY data and an X-ray crystallographic analysis. The isolated compounds (1-3) were examined for their cytotoxic activity against such human cell lines as A549, PC-3 and HCT-15.     

Structure of hemicelluloses isolated from Canavalia ensiformis and Triticum aestivum straws

Hemicellulose was extracted from horse bean and wheat straws in a yield of 5 and 9% respectively. The whole hemicellulose was hydrolysed and the molar ratio of the component monosaccharides was determined. Uronic acid, galactose, glucose, arabinose and xylose were found in both hemicelluloses. The molar ratio of the monosaccharides was determined in each of 4 fractions derived from the saccharide. The main fractions (B and C) were partially hydrolysed and an oligosaccharide containing arabinose and xylose (1:1) was isolated from both hemicelluloses. Another oligosaccharide containing xylose and glucose (2:1) was also isolated from wheat straw hemicellulose. Periodate oxidation was carried out on fractions B and C. The formic acid and the consumed periodate were determined. Each hemicellulose was subjected to Smith's degradation. Glycerol, erythrytol and compounds containing xylose and glycerol (1:1), and xylose and erythrytol (1:1) were isolated.     

The isolation and use of iron-oxidizing,moderately thermophilic acidophiles from the Collie coal mine for the generation of ferric iron leaching solution

Moderately thermophilic, iron-oxidizing acidophiles were enriched from coal collected from an open-cut mine in Collie, Western Australia. Iron-oxidizers were enriched in fluidized-bed reactors (FBR) at 60 degrees C and 70 degrees C; and iron-oxidation rates were determined. Ferrous iron oxidation by the microbiota in the original coal material was inhibited above 63;C. In addition to four iron-oxidizers, closely related to Sulfobacillus spp that had been earlier isolated from the 60 degrees C FBR, one heterotroph closely related to Alicyclobacillus spp was isolated. The Alicyclobacillus sp. isolated from the Collie coal mine tolerated a lower pH than known Alicyclobacillus spp and therefore may represent a new species. The optimum temperature for growth of the iron-oxidizing strains was approximately 50 degrees C and their maximum temperatures were approximately 60 degrees C. The FBR was adjusted to operate at 50 degrees C and was inoculated with all of the isolated iron-oxidizing strains. At 60 degrees C, an iron-oxidation rate of 0.5 g Fe(2+) l(-1) x h(-1) was obtained. At 50 degrees C, the iron-oxidation rate was only 0.3 g Fe(2+) l(-1) x h(-1). These rates compare favourably with the iron-oxidation rate of Acidianus brierleyi in shake-flasks, but are considerably lower than mesophilic iron-oxidation rates.     

Ureaplasmas and mycoplasmas in chimpanzees of various breeding capacities

Adult chimpanzees (24 male, 76 female) with low and high rates of conception were examined for ureaplasmas, arginine-metabolizing mycoplasmas and chlamydiae. Ureaplasmas were isolated from the throat of only 1 male and 1 female animal, but from the urethra of 29% of the males and from the vagina of 95% of the females. Mycoplasmas were isolated from the throat more often than were ureaplasmas, but from the genital tract with about the same frequency as ureaplasmas. The numbers of organisms, of either type, isolated from the vagina were larger than the numbers isolated from the male urethra. Chlamydiae were not isolated from any animal. The occurrence of ureaplasmas and mycoplasmas and the numbers of these organisms isolated were similar in animals with low or high rates of conception. Furthermore, no association was noted between the organisms in the lower genital tract and the occurrence of abortion and/or stillbirth.     

中华光萼苔的化学成分研究

首次对中华光萼苔(Porella chinensis(Steph.)Hat.)的化学成分进行了研究,从中分离得到四种倍半萜类化合物.通过波谱学数据并与已知化合物的数据比较,它们分别鉴定为环色烯酮(cycloeolorenone,1)、辛辣木醇(drimenol,2)、辛辣木素(drimenin,3)及α-古巴烯(α-copacne,4).通过NOESY确定了化合物1的相对构型.化合物2和4首次从该植物的配子体中分离得到.     

The glycoprotein-bound large carbohydrates from embryonal carcinoma cells carry receptors for several lectins recognizing N-acetylgalactosamine and galactose

When various lectins were mixed with radioactively labeled embryoglycan (polylactosamine-type glycoprotein-bound carbohydrates from early embryonic cells) isolated from F9 embryonal carcinoma cells and the resulting complex was precipitated with ammonium sulfate, the glycan was found to react with the following lectins: Helix pomatia agglutinin (HPA), soybean agglutinin (SBA), Sophora japonica agglutinin (SJA), and Ricinus communis agglutinin-1 (RCA-1). Furthermore, affinity chromatography on lectin-agarose revealed that receptors for Griffonia simplicifolia agglutinin-I (GS-I) were also carried by the glycan. Together with the previous finding that the glycan carries receptors for Dolichos biflorus agglutinin (DBA) and peanut agglutinin (PNA), the present result established that the glycan has receptors for a variety of lectins recognizing N-acetylgalactosamine and/or galactose in teratocarcinoma cells. Intact molecules carrying GS-1 receptors and SJA receptors were isolated from F9 cells and teratocarcinoma OTT6050 and were shown to be high-molecular weight glycoproteins similar to DBA receptors isolated from the same sources.     

Modulation of murine macrophage responses stimulated with influenza glycoproteins.

Previously it was reported that influenza virus stimulated, nonspecific resistance was largely due to its glycoproteins, hemagglutinin (HA) and neuraminidase (NA). The enhancement of natural killer cell activity was the intrinsic property of NA and HA. In the present study, the stimulatory effect of these glycoproteins on the murine peritoneal macrophages was studied. Electrophoretically purified glycoproteins, NA and HA, of influenza virus A/USSR/90/77 (H1N1) were administered intraperitoneally to C3H/HeN mice, with or without stearyl tyrosine (ST). Macrophages were isolated and were restimulated with phorbol myristate acetate. H2O2 secretion was determined by horseradish peroxidase dependent oxidation of phenol red assay. HA enhanced H2O2 secretion only in the presence of ST (60 nmol.mg-1.h-1), whereas NA alone stimulated H2O2 secretion (83 nmol.mg-1.h-1), by 6-fold over control (13 nmol.mg-1.h-1), and this stimulation was further increased (136 nmol.mg-1.h-1) in the presence of ST. Interleukin 1 (IL-1) activity was determined by using D10.G4.1 cells. There was a little stimulation of IL-1 activity (less than 1 U/mL) of macrophages isolated from HA-primed of HA+ST-primed mice restimulated with HA. On the other hand, IL-1 activity of macrophages isolated from NA-primed mice restimulated with NA significantly increased (102 U/mL) over control (less than 1 U/mL), and an additional 2-fold increase (231 U/mL) resulted when macrophages from NA+ST-primed mice were used. Tumor necrosis factor (TNF) activity was examined by using L929 cells. Negligible TNF activity was observed in macrophages isolated from either HA-primed or HA+ST-primed mice restimulated with HA.(ABSTRACT TRUNCATED AT 250 WORDS)     

Isolation and identification of yeast messenger ribonucleic acids coding for enolase, glyceraldehyde-3-phosphate dehydrogenase, and phosphoglycerate kinase.

Yeast poly(adenylic acid)-containing messenger ribonucleic acid isolated from two strains of Saccharomyces cerevisiae was fractionated by preparative polyacrylamide gel electrophoresis in the presence of formamide. Three messenger ribonucleic acids, present at high intracellular concentration, were electrophoretically eluted from the polyacrylamide gels and translated in a wheat germ cell-free extract. The in vitro synthesized polypeptides were identified by tryptic peptide analysis. Messenger ribonucleic acids coding for enolase and glyceraldehyde-3-phosphate dehydrogenase were isolated from commercially grown baker's yeast (strain F1), and messenger ribonucleic acid coding for phosphoglycerate kinase was isolated from Saccharomyces cerevisiae (ATCC 24657). Significant differences in the spectrum of abundant messenger ribonucleic acids isolated from commercially grown baker's yeast (strain F1) and strain 24657 were observed. When both strains were grown under identical conditions, however, the spectrum of messenger ribonucleic acid isolated from the cells is indistinguishable.     

Sulfate-reducing bacteria in rice field soil and on rice roots.

Rice plants that were grown in flooded rice soil microcosms were examined for their ability to exhibit sulfate reducing activity. Washed excised rice roots showed sulfate reduction potential when incubated in anaerobic medium indicating the presence of sulfate-reducing bacteria. Rice plants, that were incubated in a double-chamber (phylloshpere and rhizosphere separated), showed potential sulfate reduction rates in the anoxic rhizosphere compartment. These rates decreased when oxygen was allowed to penetrate through the aerenchyma system of the plants into the anoxic root compartment, indicating that sulfate reducers on the roots were partially inhibited by oxygen or that sulfate was regenerated by oxidation of reduced S-compounds. The potential activity of sulfate reducers on rice roots was consistent with MPN enumerations showing that H2-utilizing sulfate-reducing bacteria were present in high numbers on the rhizoplane (4.1 x 10(7) g-1 root fresh weight) and in the adjacent rhizosperic soil (2.5 x 10(7) g-1 soil dry weight). Acetate-oxidizing sulfate reducers, on the other hand, showed highest numbers in the unplanted bulk soil (1.9 x 10(6) g-1 soil dry weight). Two sulfate reducing bacteria were isolated from the highest dilutions of the MPN series and were characterized physiologically and phylogenetically. Strain F1-7b which was isolated from the rhizoplane with H2 as electron donor was related to subgroup II of the family Desulfovibrionaceae. Strain EZ-2C2, isolated from the rhizoplane on acetate, grouped together with Desulforhabdus sp. and Syntrophobacter wolinii. Other strains of sulfate-reducing bacteria originated from bulk soil of rice soil microcosms and were isolated using different electron donors. From these isolates, strains R-AcA1, R-IbutA1, R-PimA1 and R-AcetonA170 were Gram-positive bacteria which were affiliated with the genus Desulfotomaculum. The other isolates were members of subgroup II of the Desulfovibrionaceae (R-SucA1 and R-LacA1), were related to Desulforhabdus sp. (strain BKA11), Desulfobulbus (R-PropA1), or culstered between Desulfobotulus sapovorans and Desulfosarcina variabilis (R-ButA1 and R-CaprA1).     

Interaction of Escherichia coli adenosine triphosphatase with aurovertin and citreoviridin: inhibition and fluorescence studies.

Aurovertins B and D inhibited the adenosine triphosphatase (ATPase) activity of soluble Escherichia coli coupling factor ATPase (BF1) isolated from wile-type E. coli K-12. Half inhibition was obtained with 2 microns aurovertin B and 0.9 microns aurovertin D. Aurovertins B and D had no inhibitory effect on BF1 isolated from the aurovertin-resistant E. coli mutant MA12. Acetylation or saponification of aurovertin D yielded a derivative which was devoid of inhibitory effect on BF1. Citreoviridin also inhibited wild-type BF1 but with much less efficiency (half inhibition at 60 microns) than aurovertin. Citreoviridin had no effect on the aurovertin-resistant BF1. The fluorescence intensity of aurovertins B and D was markedly enhanced upon addition to purified BF1. There was no enhancement of fluorescence when the aurovertins were added to BF1 isolated from the aurovertin-resistant mutant. The fluorescence of the aurovertin-BF1 complex was enhanced by adenosine 5'-diphosphate and by low concentrations of adenosine 5'-triphosphate. The adenosine 5'-diphosphate-enhanced fluorescence of the aurovertin-BF1 complex was quenched by high concentrations of adenosine 5'-triphosphate or by MG2+. Aurovertin bound selectively to the beta subunit of BF1 isolated from wile-type cells. By complementation assays in vitro, using a reconstituted system made of subunits isolated from wild-type and aurovertin-resistant BF1, it was shown that the altered peptide in aurovertin-resistant BF1 was the beta subunit.