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951.
By inserting appropriate peptide ligands into surface loops on globular proteins, we expect to develop probes for the location, accessibility, and steric and electrostatic environment of these ligand-binding sites on their membrane-bound receptors. Three residues in a loop on the surface of E. coli alkaline phosphatase were substituted by an 18-residue peptide containing the receptor-binding segment of somatostatin-14 without significantly affecting the catalytic properties of the enzyme. This hybrid protein was then used to investigate the ligand-binding site of somatostatin receptors. Tryptic cleavage of the hybrid protein within the inserted sequence, and binding of the hybrid protein to antisomatostatin antibodies demonstrated the surface accessibility of the guest peptide. Both the wild-type enzyme and the hormone-enzyme hybrid displaced 125I-labeled somatostatin from rat brain membrane receptors only at high concentrations. However, chemical cationization of the hybrid protein, which again did not disturb the phosphatase activity, enhanced its receptor-binding potency to a level only 23 times lower than that of somatostatin itself and 280 times higher than that of the cationized wild-type protein. This alkaline phosphatase/somatostatin hybrid protein appears, therefore, to be a suitable starting point for the development of probes for the steric and electrostatic environment of the ligand-binding site of somatostatin receptors.  相似文献   
952.
The synthesis and pharmacology of 15 1-deoxy-Δ8-THC analogues, several of which have high affinity for the CB2 receptor, are described. The deoxy cannabinoids include 1-deoxy-11-hydroxy-Δ8-THC (5), 1-deoxy-Δ8-THC (6), 1-deoxy-3-butyl-Δ8-THC (7), 1-deoxy-3-hexyl-Δ8-THC (8) and a series of 3-(1′,1′-dimethylalkyl)-1-deoxy-Δ8-THC analogues (2, n=0–4, 6, 7, where n=the number of carbon atoms in the side chain−2). Three derivatives (1719) of deoxynabilone (16) were also prepared. The affinities of each compound for the CB1 and CB2 receptors were determined employing previously described procedures. Five of the 3-(1′,1′-dimethylalkyl)-1-deoxy-Δ8-THC analogues (2, n=1–5) have high affinity (Ki=<20 nM) for the CB2 receptor. Four of them (2, n=1–4) also have little affinity for the CB1 receptor (Ki=>295 nM). 3-(1′,1′-Dimethylbutyl)-1-deoxy-Δ8-THC (2, n=2) has very high affinity for the CB2 receptor (Ki=3.4±1.0 nM) and little affinity for the CB1 receptor (Ki=677±132 nM).
Scheme 3. (a) (C6H5)3PCH3+ Br, n-BuLi/THF, 65°C; (b) LiAlH4/THF, 25°C; (c) KBH(sec-Bu)3/THF, −78 to 25°C then H2O2/NaOH.  相似文献   
953.
Sedláková  Iva  Chytrý  Milan 《Plant Ecology》1999,143(1):77-87
Vegetation development in dry heathlands was studied during a six-year period following experimental burning, sod-cutting and cutting in a continental area in the southern part of the Czech Republic (Podyjí National Park). Calluna vulgaris-dominated heathlands in the area were composed of uneven-aged stands and had been unmanaged for several decades. The aim of the study was testing the applicability of Western European management systems to nature conservation in a continental area. Species composition was recorded in 8 permanent plots using 25×25 cm grids, and vegetation recovery following different treatments was analysed. Calluna recovered successfully after burning. Burning dense heathlands, which contained a large amount of woody fuel, resulted in a medium-intensity fire that destroyed moss mats and litter and exposed patches of bare ground to facilitate Calluna regeneration by seed, in addition to vegetative regrowth. On the contrary, in open heathlands with patches of herbaceous vegetation, low-intensity fires which failed to expose mineral soil were more typical. Almost all Calluna regeneration was vegetative in this case, and regrowth was slower. Heathland recovery after sod-cutting to mineral soil depended on whether or not Calluna seed germination occurred in the plot. With germination, the community developed towards heathland; without towards grassland. Cutting promoted a striking increase in grass cover, which was followed by the slow recovery of Calluna. The experiments suggest that of these management systems, burning is perhaps the most appropriate in the study area. A nature conservation management system is proposed, based on sheep grazing combined with rotational burning of restricted patches.  相似文献   
954.
Wang XS  Dong Q  Zuo JP  Fang JN 《Carbohydrate research》2003,338(22):2393-2402
S3A was a RG-I pectin isolated from Centella asiatica that contained Rha, Ara, Gal, Glc and GalA in molar ratio of 1.0:0.6:1.5:0.2:1.1 and had been found to have a backbone composed mainly of the disaccharide repeat unit, -->4)-alpha-D-GalpA-(1-->2)-alpha-L-Rhap-(1-->. Based on methylation analysis, NaIO4 oxidation, partial acid hydrolysis and lithium-treatment, the structural features were elucidated. Side chains of S3A were predominantly linked to O-4 of 1,2,4-linked alpha-L-Rhap. The side chains are comprised of arabinosyl chains, galactosyl chains, arabinogalactosyl chains and short glucosyl chains. A total of 45% Rhap in the backbone was substituted by side chains. The arabinosyl residues were mostly distributed in the arabinosyl side chains. According to the immunological results of S3A and its degraded derivatives, S3A had no immunological activity, but its derivatives had immuno-stimulating activities to some extent.  相似文献   
955.
The internal transcribed spacer (ITS) region of the nuclear ribosomal DNA from cultivated lentil (Lens culinaris subsp. culinaris) and its wild relatives was isolated and analysed for nucleotide sequence variation. Sequence divergence values ranged from no polymorphism within single species and between the cultigen and one accession of its wild progenitor (L. culinaris subsp. orientalis) to 14 base substitutions between L. nigricans and L. lamottei. Jukes and Cantor distance ranged from 0 to 1.79 %. Phylogenetic analysis confirmed the divergence of L. nigricans from all species, and the closeness of cultivated lentil to its wild progenitor, although two gene pools could possibly be identified in subsp. orientalis. Based on this study, the two recently recognized species, L. lamottei and L. tomentosus were separated from the other species. Each wild species showed peculiar autapomorphies and, in general, did not display much variation among accessions. The trees using chickpea as an outgroup formed two main clusters, one constituted by L. nigricans only and the other including the remaining taxa. Within this larger group, small subclades could be identified.  相似文献   
956.
Eight new alkaloids, the pyrido[1,2-a]azepines stemokerrin, methoxystemokerrin-N-oxide, oxystemokerrin, oxystemokerrin-N-oxide, and pyridostemin, along with the pyrrolo[1,2-a]azepines dehydroprotostemonine, oxyprotostemonine, and stemocochinin were isolated from four Stemona species together with the known compounds protostemonine, stemofoline, 2'-hydroxystemofoline, and parvistemonine. Their structures were elucidated by 1H and 13C NMR including 2D methods and two key compounds additionally by X-ray diffraction. Besides the formation of a six membered piperidine ring, additional oxygen bridges and N-oxides contributed to structural diversity. The co-occurrence of pyrrolo- and pyridoazepines suggested biosynthetic connections starting from more widespread protostemonine type precursors. Bioassays with lipophilic crude extracts against Spodoptera littoralis displayed very strong insecticidal activity for the roots of S. curtisii and S. cochinchinensis, moderate activity for S. kerrii, but only weak effects for the unidentified species HG 915. The insect toxicity was mainly caused by the accumulation of stemofoline, oxystemokerrin, and dehydroprotostemonine displaying two different modes of action. Based on the various insecticidal activities of 13 derivatives structure-activity relationships became apparent.  相似文献   
957.
The transferability of microsatellite markers developed for olive cultivars (Olea europaea L.) has been tested and confirmed in the Olea complex. Thirty two genotypes, belonging to different taxa of the genus Olea, have been analyzed with four olive SSRs. Positive amplifications at all loci were obtained in 13 taxa (at least one accession per species). Sixty seven different alleles have been detected at the four loci analyzed. Polymorphic products have been observed at the inter- and intra-species level. Some SSR loci have shown multiple amplification products in some species. The high number of unique alleles has allowed the unambiguous discrimination of most accessions. Similarity coefficients and relationships among the Olea taxa have been calculated based on SSR amplification results. The reliability of SSRs as markers for intra-species variability evaluation has been confirmed while their use to explore relationships at the inter-species level is discussed, being dependent on the locus analyzed.Communicated by H.F. Linskens  相似文献   
958.
Nineteen cattle bones from the Viking 10th and early 11th century levels in Dublin were assessed for presence of reliable genotypes from three autosomal markers. Due to the good preservational condition of the samples, it was possible to amplify and type at least two out of three of the microsatellite markers (CSRM60, HEL1 and ILSTS001) in 11 specimens. Full three-loci genotypes were obtained from a subset of seven of these samples. A comparative analysis was performed using data from the same three markers in 11 extant British, Irish and Nordic cattle breeds. Although the medieval remains displayed lower levels of diversity than the modern European breeds, the results fit within the ranges obtained from the extant populations. The results indicate a probable origin for the ancient Irish cattle as the remains group significantly more closely with breeds from the British Isles than with those from Scandinavia. The data collected indicate that microsatellites may be useful for the further study of ancient cattle.  相似文献   
959.
The paper discusses the main changes in the composition of mixed deciduous forests which occurred mostly between 5,000 and 2,000 b.p., based on selected pollen diagrams from the lowlands of Germany, Denmark and Poland, and including two pollen diagrams from varved sediments, used as reference sites, and on isopollen maps for Poland. The Ulmus retreat is shown on maps, and additional data for its pathogenic origin are presented. Corylus declines at ca. 3,500 b.p. at both reference sites, and its connection with Fagus expansion in the west and Carpinus expansion in the east is discussed. The nature of post-Atlantic transitory shrub–forest communities with dominant Corylus and Quercus is presented. Relationships between the history of Fagus and Carpinus and the development of human settlements are shown. Human impact has been admitted as one of the most important driving forces determining vegetational development since the time of fully developed Neolithic cultures. Other very important abiotic factors were the climate (particularly after 2,500 b.p.), and soil degradation.  相似文献   
960.
Shukla A  Guptasarma P 《Proteins》2004,57(3):548-557
We show that residues at the interfaces of protein-protein complexes have higher side-chain energy than other surface residues. Eight different sets of protein complexes were analyzed. For each protein pair, the complex structure was used to identify the interface residues in the unbound monomer structures. Side-chain energy was calculated for each surface residue in the unbound monomer using our previously developed scoring function.1 The mean energy was calculated for the interface residues and the other surface residues. In 15 of the 16 monomers, the mean energy of the interface residues was higher than that of other surface residues. By decomposing the scoring function, we found that the energy term of the buried surface area of non-hydrogen-bonded hydrophilic atoms is the most important factor contributing to the high energy of the interface regions. In spite of lacking hydrophilic residues, the interface regions were found to be rich in buried non-hydrogen-bonded hydrophilic atoms. Although the calculation results could be affected by the inaccuracy of the scoring function, patch analysis of side-chain energy on the surface of an isolated protein may be helpful in identifying the possible protein-protein interface. A patch was defined as 20 residues surrounding the central residue on the protein surface, and patch energy was calculated as the mean value of the side-chain energy of all residues in the patch. In 12 of the studied monomers, the patch with the highest energy overlaps with the observed interface. The results are more remarkable when only three residues with the highest energy in a patch are averaged to derive the patch energy. All three highest-energy residues of the top energy patch belong to interfacial residues in four of the eight small protomers. We also found that the residue with the highest energy score on the surface of a small protomer is very possibly the key interaction residue.  相似文献   
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