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11.
12.
Levels of katahdinone (solavetivone), lubimin, rishitin, and phytuberin, sesquiterpenoid stress metabolites of white potato (Solanum tuberosum), were monitored in tuber slices which were challenged with an extract of Phytophthora infestans and incubated under controlled atmospheres. A mixture of ethylene in air enhanced stress metabolite production. This enhancement was amplified by higher partial pressures of oxygen. Stress metabolite production was inhibited by salicylhydroxamic acid. These results suggest the involvement of cyanide-resistant respiration in the production of potato stress metabolites, compounds which may serve as phytoalexins.  相似文献   
13.
The absolute configurations of isoflavans and isoflavanquinones isolated from Cyclolobium, Dalbergia and Machaerium species were established by comparison of their ORD curves with that of (3S)-5,7,3′,4′-tetra-methoxyisoflavan and (3S)-7,4′-dimethoxyisoflavan-2′,5′-quinone, respectively. The assignments were checked by the ozonolysis of the isoflavan (?)-duartin to (R)-paraconic acid and the oxidation of isoflavans to isoflavanquinones. The PMR spectra of the dihydropyran ring of the isoflavans are discussed in terms of the preferred conformation of this ring.  相似文献   
14.
The potentiometric acid-base titration curve of fully protonated lysozyme at ionic strengths of 0.10 and 1.0 m has been performed. The stoichiometry and the pKa values of each titratable group have been determined through the linearization of titration curves. Two types of carboxylic groups with pKa values of 3.76 and 5.02, the imidazole group with pKa 7.37 and the amine group with pKa 9.63, have been identified at an ionic strength of 0.10 m at 25.0°C. The number of titratable groups found per mole of protein has been 5.12 and 5.60 for the two types of carboxylic groups, 1.13 for the imidazole group, and 3.19 for the amino groups. The endpoint of the titration of the protein obtained by this method accords quite well with the endpoint obtained by the use of Gran function applied to the excess of strong base.  相似文献   
15.
Antistasin, a 15-kDa anticoagulant protein isolated from the salivary glands of the Mexican leech Haementeria officinalis, has been shown to be a potent inhibitor of factor Xa in the blood coagulation cascade. Antistasin possesses a twofold internal homology between the N- and C-terminal halves of the molecule, suggesting a gene duplication event in the evolution of the antistasin gene. This structural feature also suggests that either or both halves of the protein may possess biological activity if expressed as separate domains. Because the N-terminal domain contains a factor Xa P1-reactive site, we chose to express this domain in an insect cell baculovirus expression system. Characterization of this recombinant half antistasin molecule reveals that the N-terminal domain inhibits factor Xa in vitro, with a K(i) of 1.7 nM.  相似文献   
16.
A method for the electron microscopic study of human peripheral lymphocytes by which very small buffy coats are obtained through centrifugation of heparinized whole blood in glass or plastic microhematocrit tubes is presented. This method is time saving and efficient, yielding well preserved material and a comparatively large number of mononuclear cells (mainly lymphocytes) in each thin section.  相似文献   
17.
Many plants pollinated by nectar-foraging animals have to maintain a balance between legitimate visitor attraction strategies and mechanisms that minimize illegitimate visits. This study investigated how floral display and neighboring species composition influences nectar robbing by hummingbirds in the tropical ornithophilous herb Heliconia spathocircinata. We tested the role of inflorescence display, flower abundance, and neighboring species in the reduction of nectar robbing in H. spathocircinata. Our results indicate that nectar robbing hummingbird activity was higher in moderately large inflorescence displays and that the frequency of nectar robbing in H. spathocircinata decreases with increased flower abundance and the presence of neighboring plant species. Neighboring non-ornithophilous plants decreased the frequency of nectar robbing in H. spathocircinata flowers to a greater extent than ornithophilous ones. These results suggest that nectar robbing hummingbirds are attracted to similar conditions that attract legitimate visitors and that spatial aggregation and mixed-species displays may represent a mechanism to dilute nectar robbing effects at an individual level.  相似文献   
18.
Volatility and thermal stability are necessary physical-chemical properties for analysing a substance by gas chromatography. A derivatization step is required before gas chromatography of benzoylecgonine (the main metabolite of cocaine). In the literature, reactions such as silylation, perfluoroalkylation or alkylation are the most frequently used to derivatize benzoylecgonine. However, they allow the formation of products sensitive to moisture or require a purification step. So, a procedure to derivatize benzoylecgonine with diazomethane before gas chromatographic analysis was evaluated. A study was performed to evaluate the efficiency of conversion of benzoylecgonine in cocaine, the necessary time for reaction and the stability of ethereal solution of diazomethane. The reaction was shown to be very fast in mild conditions and there was no need for a further purification step. When benzoylecgonine was extracted from urine by solid-phase extraction and derivatized with diazomethane, concentrations as low as 25 ng/ml could be detected using GC-MS in the full scan mode.  相似文献   
19.
Brachypodium distachyon is a promising model system for the structural and functional genomics of temperate grasses because of its physical, genetic and genome attributes. The sequencing of the inbred line Bd21 ( http://www.brachypodium.org ) started in 2007. However, a transformation method remains to be developed for the community standard line Bd21. In this article, a facile, efficient and rapid transformation system for Bd21 is described using Agrobacterium -mediated transformation of compact embryogenic calli (CEC) derived from immature embryos. Key features of this system include: (i) the use of the green fluorescent protein (GFP) associated with hygromycin selection for rapid identification of transgenic calli and plants; (ii) the desiccation of CEC after inoculation with Agrobacterium ; (iii) the utilization of Bd21 plants regenerated from tissue culture as a source of immature embryos; (iv) the control of the duration of the selection process; and (v) the supplementation of culture media with CuSO4 prior to and during the regeneration of transgenic plants. Approximately 17% of CEC produced transgenic plants, enabling the generation of hundreds of T-DNA insertion lines per experiment. GFP expression was observed in primary transformed Bd21 plants (T0) and their progeny (T1). The Mendelian inheritance of the transgenes was confirmed. An adaptor-anchor strategy was developed for efficient retrieval of flanking sequence tags (FSTs) of T-DNA inserts, and the resulting sequences are available in public databases. The production of T-DNA insertion lines and the retrieval of associated FSTs reported here for the reference inbred line Bd21 will facilitate large-scale functional genomics research in this model system.  相似文献   
20.
Infections caused by Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa foremost among them, constitute a major worldwide health problem. Bioinformatics methodologies are being used to rationally design new antimicrobial peptides, a potential alternative for treating these infections. One of the algorithms used to develop antimicrobial peptides is the Joker, which was used to design the peptide PaDBS1R6. This study evaluates the antibacterial activities of PaDBS1R6 in vitro and in vivo, characterizes the peptide interaction to target membranes, and investigates the PaDBS1R6 structure in contact with mimetic vesicles. Moreover, we demonstrate that PaDBS1R6 exhibits selective antimicrobial activity against Gram-negative bacteria. In the presence of negatively charged and zwitterionic lipids the structural arrangement of PaDBS1R6 transits from random coil to α-helix, as characterized by circular dichroism. The tertiary structure of PaDBS1R6 was determined by NMR in zwitterionic dodecylphosphocholine (DPC) micelles. In conclusion, PaDBS1R6 is a candidate for the treatment of nosocomial infections caused by Gram-negative bacteria, as template for producing other antimicrobial agents.  相似文献   
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