Volatile Components of Quince Fruit (Cydonia oblonga Mill.)

Steam distilled oil of quince fruit (Cydonia oblonga Mill.=C. vulgaris Pers., marmelo in Japanese) was analyzed by gas chromatography and gas chromatography-mass spectrometry. Sixty-two compounds, 2 hydrocarbons, 13 esters, 11 alcohols, 11 aldehydes, 11 ketones, 5 lactones and 9 miscellaneous compounds, were identified. Of them, the chemical structures of two new oxide compounds, trans- and cis-3-methyl-5-[(E)-3′-methyl-13′-butadien-1′-yl]tetrahydrofuran, were elucidated by instrumental analyses.     

A Note on the Limitations of Identifying Soft Rot Erwinias by Temperature Tolerances and Sensitivity to Erythromycin on a Pectate Medium

Forty-five erwinia strains from 14 different hosts have been tested lor growth on a crystal violet pectate medium with or without 70 μ/ml erythromycin at 27, 33.5 and 37°C. On the basis of positive data in the literature it was tried to identify the strains as Erwnia carotovora subsp. atroseptica, E. carotovora subsp. carotovora or E. chrysanthemi using this method. It appeared that relatively few strains reacted differentially, apparently due to strain variation (all strains were identified also by biochemical and physiological tests). The conclusion is that for a reliable identification of field isolates of soft rot crwinias other (biochemical, physiological and serological) methods have to be applied.     

Simultaneous determination of triamcinolone acetonide and hydrocortisone in human plasma by high-performance liquid chromatography

A validated HPLC method for the simultaneous determination of triamcinolone acetonide and hydrocortisone has been established to monitor the plasma levels of the compounds in healthy volunteers following intramuscular (i.m.) administration of triamcinolone acetonide. Plasma (1.0 ml) was extracted with dichloromethane after addition of the internal standard, fluocortolone. The compounds were separated using a LiChrospher RP 18 column and detected by UV absorbance. Specificity, linearity, as well as the repeatability, intermediate precision and accuracy of the method were established. The limit of quantification was 0.6 ng/ml for triamcinolone acetonide (C.V.=8.7%, R.E.=2.6%, n=6) and 2.0 ng/ml for hydrocortisone (C.V.=8.3%, R.E.=2.8%, n=6). Data on the stability of triamcinolone acetonide in human plasma are presented. Recovery of the compounds and the internal standard have been studied. The results of quality control samples (n=126) determined during routine analysis of volunteer samples are described. Plasma levels of triamcinolone acetonide after i.m. administration of 40 mg of triamcinolone acetonide are presented.     

贝类壳-体质量比和静水沉降特性的相关性

软体动物贝壳表型是适应性进化的结果,为探究贝壳形态特性与软体动物随水流扩散功能的相关性,实验选取9种具代表性的腹足纲和双壳纲贝类作为研究对象,研究了其成体的壳-体质量比与静水沉降特性及二者的相关性。结果表明：体质量的变异系数（C.V.=1.11）大于壳长（C.V.=0.67）、壳宽（C.V.=0.54）的变异系数;壳-体质量比具较强的种属特异性,种内变异系数小（C.V.=0.20）,而种间差异极显著（P < 0.01）。受试贝类的平均壳-体质量比为0.32±0.13,陆生贝类的壳-体质量比显著小于水生贝类（P < 0.01）,壳-体质量比最大的河蚬（0.54±0.06）是最小的非洲大蜗牛（0.17±0.04）的3倍,福寿螺的壳-体质量比为0.50±0.06。双壳类和螺类的沉降行为差异明显,只有中华园田螺、铜锈环棱螺和河蚬不能在静水中漂浮;沉降速度最大的河蚬（（24.99±4.22）cm/s）是最小的椭圆萝卜螺（（4.13±0.96）cm/s）的6倍,入侵种非洲大蜗牛（（18.30±3.64）cm/s）和福寿螺（（21.77±5.23）cm/s）与土著种铜锈环棱螺（（19.48±3.14）cm/s）和中华园田螺（（21.44±3.92）cm/s）在沉降速度上无显著差异（P < 0.01）。成体贝类的壳-体质量比与沉降速度（R²=0.28）、沉降加速度（R²=0.39）之间存在一定的相关性。在此基础上,贝类沉降特性随生活史变化的研究将进一步揭示贝类进化与种群扩散中的生态水力学作用。     

Biochemical characterisation of Nicotiana plumbaginifolia auxotrophs that require branched-chain amino acids

The biochemical lesions in six amino acid-requiring auxotrophic lines of Nicotiana plumbaginifolia have been investigated, by means of feeding experiments with [¹⁴C] and unlabelled substrates, and enzyme analysis. Three of the lines require isoleucine for growth, are unable to synthesise 2-oxobutyrate in vivo and have no detectable threonine dehydratase (E.C.4.2.1.16) in vitro. The other three lines require (isoleucine + valine), accumulate [¹⁴C] dihydroxymethylvalerate when fed [¹⁴C]-L-threonine, and contain no detectable dihydroxyacid dehydratase (E.C.4.2.1.9).     

Evaluation of different PCR-based approaches for the identification and typing of environmental enterococci

The aim of the work was the evaluation of different PCR-based methods to found an appropriate identification and typing strategy for environmental enterococci. Environmental enterococci were isolated mainly from surface- and waste-waters. Species identification was provided by combination of phenotypic (Micronaut System, Merlin) and molecular detection methods (fluorescent ITS-PCR, ddl-PCR, REP-PCR, AFLP). Very similar results were observed among molecular methods, however several discrepancies were recognized during comparison of molecular and biochemical identification. Seven enterococcal species (E. faecium, E. hirae, E. casseliflavus, E. mundtii, E. faecalis, E. durans and E. gallinarum) were identified within 166 environmental isolates. The results obtained in this work attest the importance of PCR-based methods for identification and typing of environmental enterococci. The fluorescent ITS-PCR (fITS-PCR) showed the best results in order to identify the enterococci strains, the method used the automated capillary electrophoresis to separate the PCR products in a very rapid and precise way. The AFLP method was suitable to identify and characterize the isolates, while the REP-PCR can be used for species identification.     

Electrophoretic Analysis of Endonuclease-Generated Fragments of k-DNA,of Esterase Isoenzymes,and of Surface Proteins as Aids for Species Identification of Insect Trypanosomatids1

In order to verify the applicability of biochemical methods for species identification of Trypanosomatidae, 13 species of monoxenic trypanosomatids plus the heteroxenous Trypanosoma cruzi were comparatively analyzed by three different biochemical methods. Insect trypanosomatids examined were: Crithidia acanthocephali, C. fasciculata (three varieties), C. luciliae luciliae, C. luciliae thermophila, C. deanei, C. oncopelti, Herpetomonas muscarum muscarum, H. megaseliae, H. samuelpessoai, H. mariadeanei, Leptomonas seymouri, L. collosoma, L. samueli, and Blastocrithidia culicis. Also included in the survey were aposymbiotic strains of C. deanei and C. oncopelti. Methods used were: electrophoretic profiling of endonuclease-generated fragments of k-DNA, esterase isoenzymes profiling, and polyacrylamide-gel electrophoresis (SDS-PAGE) of radioiodinated cell surface proteins. Interspecific but not intraspecific differences were detected by all three methods among the 13 monoxenic species examined. Thus, it is concluded that these methods can be successfully used, in addition to classical criteria, for species identification of insect trypanosomatids.     

Electrophoretic Isoenzyme Patterns of Entamoeba histolytica and Entamoeba chattoni in a Primate Survey1

Stocks of Entamoeba histolytica grown in a monoxenic culture system from the feces of nonhuman primates are compared with the eleven zymodemes of E. histolytica so far demonstrated from man. In a similar fashion, Entamoeba chattoni has also been grown and identified. Both E. histolytica and E. chattoni have been demonstrated in keepers of the primate collections. Comparisons have been made using the electrophoretic patterns of three enzymes: glucosephosphate isomerase [(GPI) E.C.5.3.1.9], phosphoglucomutase [(PGM) E.C.2.7.5.1], and L-malate—NADP⁺ oxidoreductase (oxaloacetate-decarboxylating) [(ME) E.C. 1.1.1.40]. Enzyme patterns of E. histolytica from the apes were found to be identical with three of those already demonstrated from man. The enzyme pattern of E. chattoni was distinctly different from that of any of the E. histolytica zymodemes. Other protozoa found in the single fecal sample examined from each subject are also listed.     

The influence of coal ashes and soil dust on the activity of two selected hydrolytic enzymes in the liver of laboratory animals

The effect of a single intratracheal application of the electroenergetic ashes and the soil dust on the liver cell was studied by means of the histoenzymatic methods for the determination of the activities of chosen hydrolases (AcP, E.C. 3.1.3.2. and ATP, E.C. 3.6.1.). The degree of the histochemical changes in hepatocytes depended on the kind of the applied dust and the trace elements' content in it. The activity's increase of the acid phosphatase and ATP-ase in each experimental group was observed, especially in the preparations collected from the animals were given the electroenergetic ashes.     

Amidase and urease activities in plants

Summary Foliar fertilization has received considerable attention in recent years. Because of the importance of amides and urea as N sources, this work was carried out to study the enzymes that catalyze the hydrolysis of these compounds in plant leaves. The methods developed for assay of these enzymes in plants involve determination by steam distillation of the NH₄ ⁺–N produced by amidase or urease activity when plant materials are incubated at 37°C with buffered (0.1M THAM pH 8.0) amide solution or buffered (0.1M THAM pH 7.5) urea solution, respectively. Amidase and urease were detected in 21 diverse plants in the families of Gramineae and Leguminosae. Results showed that amidase and urease have optimum activities at buffer pH values of 8.0 and 7.5, respectively. Both amidase and urease activities were decreased significantly upon freezing or air-drying of plant samples before enzyme assay. These differences were proportional to the original activities of fresh plant materials. Studies on the effect of temperature on amidase and urease activities showed that these enzymes are inactivated at temperatures above 60 and 70°C, respectively. The energy of activation of the reaction catalyzed by amidase and urease in plants, expressed in kJ·mole^–1, ranged from 44.0 to 51.2 (avg.=47.1) and from 43.1 to 56.5 (avg.=51.2) when formamide and urea were used as substrates, respectively. The apparent K_m constants of these enzymes varied among the plant samples studied. By using the Lineweaver-Burk plot, the K_m values for amidase when formamide was used as a substrate ranged from 2.0 to 9.4 (avg.=5.8 mM) and for urease ranged from 0.4 to 1.6 (avg.=0.8 mM). The V_max values of 7 plant samples, expressed in g of NH₄ ⁺–N produced/0.1 g of plant materials/2h, ranged from 137 to 514 for amidase and from 29 to 123 for urease. The importance of these enzymes in application of amides and urea to plant leaves is discussed.     

Enterohaemorrhagic Escherichia coli O157:H7 Shiga‐like toxin 1 is required for full pathogenicity and activation of the p38 mitogen‐activated protein kinase pathway in Caenorhabditis elegans

Enterohaemorrhagic Escherichia coli (EHEC) causes life‐threatening infections in humans as a consequence of the production of Shiga‐like toxins. Lack of a good animal model system currently hinders in vivo study of EHEC virulence by systematic genetic methods. Here we applied the genetically tractable animal, Caenorhabditis elegans, as a surrogate host to study the virulence of EHEC as well as the host immunity to this human pathogen. Our results show that E. coli O157:H7, a serotype of EHEC, infects and kills C. elegans. Bacterial colonization and induction of the characteristic attaching and effacing (A/E) lesions in the intact intestinal epithelium of C. elegans by E. coli O157:H7 were concomitantly demonstrated in vivo. Genetic analysis indicated that the Shiga‐like toxin 1 (Stx1) of E. coli O157:H7 is a virulence factor in C. elegans and is required for full toxicity. Moreover, the C. elegans p38 mitogen‐activated protein kinase (MAPK) pathway, anevolutionarily conserved innate immune and stress response signalling pathway, is activated in the regulation of host susceptibility to EHEC infection in a Stx1‐dependent manner. Our results validate the EHEC–C. elegans interaction as suitable for future comprehensive genetic screens for both novel bacterial and host factors involved in the pathogenesis of EHEC infection.     

Energy exploitation in Chirocephalus croaticus (Steuer, 1899) (Crustacea: Anostraca): survival strategy in an intermittent lake

Electron transport system (ETS) activity was measured in Chirocephalus croaticus from the intermittent lake, Petelinjsko Jezero. The ETS activities were measured at 5, 10, 15, 20, and 25 °C, and were studied separately in juveniles, females and males. Juveniles had significantly higher activity than adults at a standard temperature of 20 °C. The mass-specific ETS activity decreased with increasing size of the animals; the value b was 0.787. Respiration rates (R) were determined at 20 °C and the ratio ETS/R (±standard deviation) for C. croaticus was 1.43±0.46 (n=38). ETS activity increased with temperature. Females had higher Q₁₀ than males in higher temperature range (t-test; t=2.50; d.f.=8; p<0.05). Activation energy E_a was higher for females than males (t-test; t=2.35; d.f.=8; p<0.05). Females exhibited lower ETS activity than males over the lower temperature range, but their ETS could function more efficient at higher temperature.     

Comparison of two chromogenic media and evaluation of two molecular based identification systems for Enterobacter sakazakii detection

Background  

Enterobacter sakazakii is a foodborne pathogen that has been associated with sporadic cases and outbreaks causing meningitis, necrotizing enterocolitis and sepsis especially in neonates. The current FDA detection method includes two enrichment steps, the subculturing of the second enrichment broth on a selective agar (VRBG), a further subculturing of selected grown colonies on TSA and the subsequent biochemical identification of yellow-pigmented colonies by API20E. However, there is a strong need for simplified methods for isolation and identification of E. sakazakii. In this study, two chromogenic media, which allow to indicate presumptive E. sakazakii colonies by the alpha glucosidase activity, as well as a newly developed 1,6-alpha-glucosidase based conventional PCR assay and a rRNA oligonucleotide probe based commercial test system for identification of presumptive E. sakazakii were evaluated on 98 target and non-target strains. The methods were compared with respect to specifiCity aspects.     

Analysis of the rnc locus of Coxiella burnetii

A 3.2 kb EcoRI genomic DNA fragment of Coxiella burnetii was isolated by virtue of Its ability to suppress mucoidy in Eschertchia coli. Nucleotide sequence analysis revealed the presence of the genes homologous to rnc, era and recO of E. coli. Suppression of capsule synthesis, measured by β-galactosidase expression in Ion cps-lac fusion strains of E. coli, is caused by gene-dosage effects of the plasmid-borne rnc genes of either C. burnetii or E. coli. The rnc gene of C. burnetii complemented rn– E. coli hosts for lambda plaque morphology and stimulation of lambda N gene expression. We also demonstrated heterologous complementation of an E coli strain defective for the expression of Era, an essential protein in E. coli, using the plasmid-borne C. burnetii era. Under the control of the bacteriophage lambda P_L promoter, this 3.2 kb EcoRI DNA fragment directed the synthesis in E. coli of three proteins with approximate molecular masses of 35,27 and 25 kDa. Antibodies against purified E. coli Era protein cross-reacted with the 35 kDa protein of C. burnetii on Western blots.     

Structural-metabolic characteristics of acute hepatic failure]

Basing on early autopsy data, the study was made of structural and metabolic characteristics of the liver in marked intoxication and blood loss. Histologic, histoenzymatic and biochemical methods were used to evaluate morpho- and pathogenesis of acute liver failure arising in the above conditions.     

Cranial capacity in hominid evolution

We present an analysis of cranial capacity of 118 hominid crania available from the literature. The crania belong to both the genusAustralopithecus andHomo and provide a clear outline of hominid cranial evolution starting at more than 3 million years ago. Beginning withA. afarensis there is a clear increase in both absolute and relative brain size with every successive time period.H.s. neandertal has an absolutely and relatively smaller brain size (1412cc, E.Q.=5.6) than fossil modernH.s. sapiens (1487cc, E.Q.=5.9). Three evolutionary models of hominid brain evolution were tested: gradualism, punctuated equilibrium, and a mixed model using both gradualism and punctuated equilibrium. Both parametric and non-parametric analyses show a clear trend toward increasing brain size withH. erectus and a possible relationship within archaicH. sapiens. An evolutionary stasis in cranial capacity could not be refuted for all other taxa. Consequently, the mixed model appears to more fully explain hominid cranial capacity evolution. However, taxonomic decisions could directly compromise the possibility of testing the evolutionary mechanisms hypothesized to be operating in hominid brain expansion.     

丙型肝炎病毒依赖于RNA的RNA聚合酶(RdRp)研究进展

由于缺乏合适的HCV感染细胞模型,严重制约了HCV复制,特别是HCV复制的关键因子依赖于RNA的RNA聚合酶(RdRp)的研究.对HCV序列比较分析并通过异源表达证明NS5B是HCV复制的RdRp.NS5B C端疏水性氨基酸区域以及NS5B与细胞膜形成复合体等影响NS5B溶解性.在合适的反应条件下NS5B可以多种RNA分子为模板催化RNA复制,特别是能有效复制HCV全长(+)RNA.高浓度GTP激活HCV RdRp活性.NS5B N/C端缺失突变和保守性A、B、C区中的点突变影响RdRp活性,但D区345位精氨酸突变为赖氨酸时RdRp活性明显升高.HCV RdRp的发现及其功能研究为HCV药物研究提供了新型靶标.     

Dissection of the conformational cycle of the multidrug/lipidA ABC exporter MsbA

Recent crystal structures of the multidrug ATP‐binding cassette (ABC) exporters Sav1866 from Staphylococcus aureus, MsbA from Escherichia coli, Vibrio cholera, and Salmonella typhimurium, and mouse ABCB1a suggest a common alternating access mechanism for export. However, the molecular framework underlying this mechanism is critically dependent on assumed conformational relationships between nonidentical crystal structures and therefore requires biochemical verification. The structures of homodimeric MsbA reveal a pair of glutamate residues (E208 and E208′) in the intracellular domains of its two half‐transporters, close to the nucleotide‐binding domains (NBDs), which are in close proximity of each other in the outward‐facing state but not in the inward‐facing state. Using intermolecular cysteine crosslinking between E208C and E208C′ in E. coli MsbA, we demonstrate that the NBDs dissociate in nucleotide‐free conditions and come close on ATP binding and ADP·vanadate trapping. Interestingly, ADP alone separates the half‐transporters like a nucleotide‐free state, presumably for the following catalytic cycle. Our data fill persistent gaps in current studies on the conformational dynamics of a variety of ABC exporters. Based on a single biochemical method, the findings describe a conformational cycle for a single ABC exporter at major checkpoints of the ATPase reaction under experimental conditions, where the exporter is transport active. Proteins 2010. © 2010 Wiley‐Liss, Inc.     

Effect of pH on growth and biochemical responses of <Emphasis Type="Italic">Dunaliella bardawil</Emphasis> and <Emphasis Type="Italic">Chlorella ellipsoidea</Emphasis>

The goal of the present investigation was to study the effect of pH on growth and biochemical responses of Dunaliella bardawil and Chlorella ellipsoidea when exposed to different pH values. The two tested microalgae could grow in a wide range of pH (4–9 for D. bardawil and 4–10 for C. ellipsoidea). The dry weight gain and the biochemical components of D. bardawil were greatly enhanced at pH 7.5. In contrast, dry weight and carbohydrate content of C. ellipsoidea attained their maximum values at the alkaline pH. On the other hand, the protein content of C. ellipsoidea recorded its highest value at pH 4, while the pigment content of the same alga was highest at pH 4, 6, and 7.5 and decreased at alkaline pH. Both pH 6 and pH 9 stimulated the accumulation of β-carotene, vitamin E and vitamin C in D. bardawil, with the highest values of the three compounds recorded at pH 9. In the case of C. ellipsoidea, β-carotene content increased at pH 6 and pH 10 as compared with the control, but the amount of β-carotene was much higher at pH 6 than at pH 10. Vitamin E content was higher in C. ellipsoidea cells at pH 10 than at pH 6. Both pH 6 and pH 10 caused a significant decline in vitamin C content of C. ellipsoidea.     

Sex pheromone discrimination by male aphids of a biotype of Schizaphis graminum

The ability of male aphids of two different biotypes (C & E) of the greenbug, Schizaphis graminum (Rondani), to distinguish ovipara-produced sex pheromone of their own biotype from that of the other biotype was examined using an arena olfactometer. Biotype E males showed a strong preference for biotype E oviparae; the preference of biotype C males for biotype C oviparae was less marked. These behavioral findings indicate a potential biochemical reproductive isolating mechanism for these biotypes.

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Unterschiedung der weiblichen geschlechtspheromone zweier biotypen der aphide Schizaphis graminum durch die männchen

Zusammenfassung Die Reaktion der Männchen von zwei Biotypen (C & E) der Aphide Schizaphis graminum auf die Geschlechtspheromone der Oviparen ihres eigenen und des anderen Biotyps wurde mit Hilfe einer Olfaktometer-Arena untersucht. Biotyp E Männchen zeigten eine starke Präferenz für Biotyp E Ovipare, während die Präferenz von Biotyp C Männchen für Biotyp C Ovipare weniger stark ausgeprägt war. Diese Resultate deuten auf einen möglichen verhaltensbiologischen Isolationsmechanismus der beiden Biotypen dieser Blattlausart in der Natur hin.