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171.
172.
G. DAUBE, B. CHINA, P. SIMON, K. HVALA AND J. MAINIL. 1994. The strains of Clostridium perfringens are classified according to major toxins produced. Classically, this determination involves the seroneutralization of their lethal effect in mice. However, this method requires specific antisera and a large number of mice. In this work, a new typing method was developed based on the amplification of toxin genes by polymerase chain reaction (PCR). By combination of several pairs of primers, the toxinotype of a Cl. perfringens strain was determined by looking at the pattern of bands on an agarose gel electrophoresis. This mixture contained primers amplifying simultaneously a part of α-toxin, α-toxin, β-toxin and enterotoxin genes. In order to distinguish between toxinotype A and E, the *** l -toxin gene fragment must be amplified in a separate PCR reaction. Moreover, with the primers combination, in most cases, a PCR product corresponding to the α-toxin gene was obtained from direct enrichments of animal intestinal contents. 相似文献
173.
M. Schulz M. Bayer H. White H. Gü nther H. Simon 《Biocatalysis and Biotransformation》1994,10(1):25-36
Crude extracts of Clostridium thermoaceticum DSM 521 contain various AMAPORs (artificial mediator accepting pyridine nucleotide oxidoreductases). The specific activities of this mixture of AMAPORs is about 8-9 U mg-1 protein (µmoles mg-1 min-1) for NADPH and 3-4 U mg-1 protein for NADH formation with reduced methylviologen (MV++) as electron donor. These AMAPOR-activities are only slightly oxygen sensitive. The reoxidation of NADPH and NADH with carboxamido-methylviologen is catalysed by crude extracts with 2.0 and 1.6 U mg-1 protein, respectively. The same crude extracts also catalyse the dehydrogenation of reduced pyridine nucleotides with suitable quinones such as anthraquinone-2,6-disulphonate. The reduced quinone can be reoxidised by dioxygen.
The Km-values of these enzymes for the pyridine nucleotides and also for the artificial electron mediators are in a suitable range for preparative transformations.
Furthermore the crude extract of C. thermoaceticum contains about 2.5 U mg-1 protein of an NADP+-dependent formate dehydrogenase (FDH), which is suitable for NADPH and/or MV++ regeneration. The regeneration of MV++ with FDH and formate as electron donor proceeds with a specific activity of about 5 U mg-1 protein of the crude extract. The reduced viologen in turn reduces NAD(P)+ by AMAPOR. The formate dehydrogenase is sensitive to oxygen.
Examples of compounds which have been prepared by combination of AMAPORs or formate dehydrogenase with an oxidoreductase are: (S)-3-hydroxycarboxylates, esters of (S)-3-hydroxycarboxylates, (1R,2S)-1-hydroxypropane-tricarboxylate (Ds-(+)-isocitrate), Ls-(-)-isocitrate and 6-phosphogluconate. 相似文献
The Km-values of these enzymes for the pyridine nucleotides and also for the artificial electron mediators are in a suitable range for preparative transformations.
Furthermore the crude extract of C. thermoaceticum contains about 2.5 U mg-1 protein of an NADP+-dependent formate dehydrogenase (FDH), which is suitable for NADPH and/or MV++ regeneration. The regeneration of MV++ with FDH and formate as electron donor proceeds with a specific activity of about 5 U mg-1 protein of the crude extract. The reduced viologen in turn reduces NAD(P)+ by AMAPOR. The formate dehydrogenase is sensitive to oxygen.
Examples of compounds which have been prepared by combination of AMAPORs or formate dehydrogenase with an oxidoreductase are: (S)-3-hydroxycarboxylates, esters of (S)-3-hydroxycarboxylates, (1R,2S)-1-hydroxypropane-tricarboxylate (Ds-(+)-isocitrate), Ls-(-)-isocitrate and 6-phosphogluconate. 相似文献
174.
Developmental regulation of hexosamine biosynthesis by protein phosphatases 2A and 2C in Blastocladiella emersonii. 下载免费PDF全文
L C Etchebehere M N Simon R B Campanh P D Zapella M Vron J C Maia 《Journal of bacteriology》1993,175(16):5022-5027
Extracts of the aquatic fungus Blastocladiella emersonii were found to contain protein phosphatases type 1, type 2A, and type 2C with properties analogous to those found in mammalian tissues. The activities of all three protein phosphatases are developmentally regulated, increasing during sporulation, with maximum level in zoospores. Protein phosphatases 2A and 2C, present in zoospore extracts, catalyze the dephosphorylation of L-glutamine:fructose-6-phosphate amidotransferase (EC 2.6.1.16, amidotransferase), a key regulatory enzyme in hexosamine biosynthesis. The protein phosphatase inhibitor okadaic acid induces encystment and inhibits germ tube formation but does not affect the synthesis of the chitinous cell wall. These results strongly suggest that phosphatase 2C is responsible for the dephosphorylation of amidotransferase in vivo. This dephosphorylation is inhibited by uridine-5'-diphospho-N-acetylglucosamine, the end product of hexosamine synthesis and the substrate for chitin synthesis. This result demonstrates a dual role of uridine-5'-diphospho-N-acetylglucosamine by inhibiting the activity of the phosphorylated form of amidotransferase and by preventing its dephosphorylation by protein phosphatases. 相似文献
175.
Mortimer M. Civan Jonathan Robbins Simon Broad Enrique Rozengurt David A. Brown 《The Journal of membrane biology》1993,133(1):51-59
Summary Differentiated neuroblastoma cells exhibit both the delayed rectifier potassium current (I
K) and the M-current (I
M). The present study was designed to determine the roles of protein kinase C (PKC) and of the calmodulin-binding protein 80K/MARCKS, a prominent substrate for PKC and possible regulator of these currents. Neuroblastoma x glioma (NG108-15) hybrid cells transfected with m1 muscarinic receptors were grown with 1% fetal bovine serum (FBS) without the prostaglandin E1 (PGE1) and isobutylmethylxanthine (IBMX) usually added in preparation for electrophysiological studies. Under these conditions, the usual pleomorphism was largely abolished, leaving two populations of small cells with stellate and spherically symmetrical geometries. Whole-cell patch clamping indicated that the two cell types had identical electrophysiological properties, displaying: I
k, a small current through a T-like Ca2+ channel, and no M-current.Stimulation with carbachol shifted the distribution of cells to a more stellate morphology within 24 hr and later (after 48 hr) reduced the PKC substrate 80K/MARCKS by 22±7%. In contrast to the stimulation of I
k observed with cardiac cells, PKC activation produced only a small inhibition of I
k, which was independent of carbachol pretreatment. Thus, PKC and 80K/MARCKS can be dissociated from the regulation of I
k in neuroblastoma cells.Supported in part by research grants from the National Institutes of Health (DK-40145 and EY-08343) and from the U.K. Medical Research Council.We thank Dr. Peter J. Parker for his generous gift of PKC, and Yvonne Vallis for her skillful assistance with the cultures and harvesting of the NG108-15 transfected cells. 相似文献
176.
Simon Y. C. Wong Geoffrey R. Guile Thomas W. Rademacher Raymond A. Dwek 《Glycoconjugate journal》1993,10(3):227-234
Glycopeptides can be valuable tools in determining the influence of carbohydrate moieties on the intrinsic properties of glycoproteins. However, glycopeptides of sufficient quantity and purity are as yet not readily available from biological sources. The chemical coupling of a -glycosylamino group of an unprotected carbohydrate with an activated aspartic acid residue of an unprotected peptide is a simple method for synthesizing asparagine-linked glycopeptides. In this report we demonstrate that the use of this method is not restricted to -glycosylamines of simple monosaccharides or short aspartic acid-containing pentapeptides. This is illustrated by the syntheses of several glycopentapeptides containingN,N-diacetylchitobiose, a glutamine-linked glycopentapeptide containing a biantennary complex oligosaccharide, and glycosylated variants of two analogs of a polypeptide hormone, atriopeptin, containingN,N-diacetylchitobiose.Abbreviations Ac
acetyl
- Bzl
benzyl
- DMF
dimethylformamide
- Fmoc
9-fluorenylmethoxycarbonyl
- Fuc
fucose
- Gal
galactose
- GlcNAc
N-acetylglucosamine
- HBTU
O-benzotriazol-1-yl-N,N,N,N-tetramethyluroniumhexa-fluorophosphate
- HOBt
1-hydroxybenzotriazole
- Man
mannose
-
m/z
mass/charge
- NMR
nuclear magnetic resonance
- Xyl
xylose
- Z
benzyloxycarbonyl; unless otherwise specified, amino acids are abbreviated using their one-letter codes. 相似文献
177.
Jörg Müller Oliver Mitesser Marc W. Cadotte Fons van der Plas Akira S. Mori Christian Ammer Anne Chao Michael Scherer-Lorenzen Petr Baldrian Claus Bässler Peter Biedermann Simone Cesarz Alice Claßen Benjamin M. Delory Heike Feldhaar Andreas Fichtner Torsten Hothorn Claudia Kuenzer Marcell K. Peters Kerstin Pierick Thomas Schmitt Bernhard Schuldt Dominik Seidel Diana Six Ingolf Steffan-Dewenter Simon Thorn Goddert von Oheimb Martin Wegmann Wolfgang W. Weisser Nico Eisenhauer 《Global Change Biology》2023,29(6):1437-1450
Intensification of land use by humans has led to a homogenization of landscapes and decreasing resilience of ecosystems globally due to a loss of biodiversity, including the majority of forests. Biodiversity–ecosystem functioning (BEF) research has provided compelling evidence for a positive effect of biodiversity on ecosystem functions and services at the local (α-diversity) scale, but we largely lack empirical evidence on how the loss of between-patch β-diversity affects biodiversity and multifunctionality at the landscape scale (γ-diversity). Here, we present a novel concept and experimental framework for elucidating BEF patterns at α-, β-, and γ-scales in real landscapes at a forest management-relevant scale. We examine this framework using 22 temperate broadleaf production forests, dominated by Fagus sylvatica. In 11 of these forests, we manipulated the structure between forest patches by increasing variation in canopy cover and deadwood. We hypothesized that an increase in landscape heterogeneity would enhance the β-diversity of different trophic levels, as well as the β-functionality of various ecosystem functions. We will develop a new statistical framework for BEF studies extending across scales and incorporating biodiversity measures from taxonomic to functional to phylogenetic diversity using Hill numbers. We will further expand the Hill number concept to multifunctionality allowing the decomposition of γ-multifunctionality into α- and β-components. Combining this analytic framework with our experimental data will allow us to test how an increase in between patch heterogeneity affects biodiversity and multifunctionality across spatial scales and trophic levels to help inform and improve forest resilience under climate change. Such an integrative concept for biodiversity and functionality, including spatial scales and multiple aspects of diversity and multifunctionality as well as physical and environmental structure in forests, will go far beyond the current widely applied approach in forestry to increase resilience of future forests through the manipulation of tree species composition. 相似文献
178.
Variation in tooth crown morphology plays a crucial role in species diagnoses, phylogenetic inference, and the reconstruction of the evolutionary history of the primate clade. While a growing number of studies have identified developmental mechanisms linked to tooth size and cusp patterning in mammalian crown morphology, it is unclear (1) to what degree these are applicable across primates and (2) which additional developmental mechanisms should be recognized as playing important roles in odontogenesis. From detailed observations of lower molar enamel–dentine junction morphology from taxa representing the major primate clades, we outline multiple phylogenetic and developmental components responsible for crown patterning, and formulate a tooth crown morphology framework for the holistic interpretation of primate crown morphology. We suggest that adopting this framework is crucial for the characterization of tooth morphology in studies of dental development, discrete trait analysis, and systematics. 相似文献
179.
Sleep and Biological Rhythms - 相似文献
180.
Bram Danneels Monique Blignaut Guillaume Marti Simon Sieber Peter Vandamme Marion Meyer Aurélien Carlier 《Environmental microbiology》2023,25(2):454-472
The symbioses between plants of the Rubiaceae and Primulaceae families with Burkholderia bacteria represent unique and intimate plant–bacterial relationships. Many of these interactions have been identified through PCR-dependent typing methods, but there is little information available about their functional and ecological roles. We assembled 17 new endophyte genomes representing endophytes from 13 plant species, including those of two previously unknown associations. Genomes of leaf endophytes belonging to Burkholderia s.l. show extensive signs of genome reduction, albeit to varying degrees. Except for one endophyte, none of the bacterial symbionts could be isolated on standard microbiological media. Despite their taxonomic diversity, all endophyte genomes contained gene clusters linked to the production of specialized metabolites, including genes linked to cyclitol sugar analog metabolism and in one instance non-ribosomal peptide synthesis. These genes and gene clusters are unique within Burkholderia s.l. and are likely horizontally acquired. We propose that the acquisition of secondary metabolite gene clusters through horizontal gene transfer is a prerequisite for the evolution of a stable association between these endophytes and their hosts. 相似文献