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A cell-based high-throughput screen that assessed the cellular stability of a tumor suppressor protein PDCD4 (Programmed cell death 4) was used to identify a new guanidine-containing marine alkaloid mirabilin K (3), as well as the known compounds mirabilin G (1) and netamine M (2). The structures of these tricyclic guanidine alkaloids were established from extensive spectroscopic analyses. Compounds 1 and 2 inhibited cellular degradation of PDCD4 with EC50 values of 1.8 μg/mL and 2.8 μg/mL, respectively. Mirabilin G (1) and netamine M (2) are the first marine natural products reported to stabilize PDCD4 under tumor promoting conditions.  相似文献   
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Lawsonia intracellularis is an intracellular organism that causes proliferative enteritis in pigs. This bacterium is difficult to culture, and antemortem demonstration of the microbe is therefore often performed on fecal samples by polymerase chain reaction (PCR). Polymerase chain reaction is sensitive and specific, but inhibitory factors in feces might cause false-negative results. This article describes the construction and use of an internal standard, a mimic. The mimic is amplified by the same primers as those used for L. intracellularis DNA and thus could indicate false-negative results in clinical samples. The amplicon was clearly visible when as few as 10 mimic molecules were added per amplification reaction and when no inhibitors werepresent. When fecal samples were spiked with the mimic, the detection limit was 10(2) molecules per PCR. Sixty clinical samples, 20 from wild boars, 20 from growing pigs with diarrhea, and 20 from pigs without diarrhea, were prepared by a boiling procedure and subjected to PCR together with 10(3) mimic molecules. Nine samples were positive, of which 7 originated from pigs with diarrhea and 2 from pigs without diarrhea. In 14 samples from wild boars, in 8 samples from pigs without diarrhea, and in 3 samples from pigs with diarrhea, neither the mimic nor the target DNA was visible. This indicated the presence of inhibitors in these samples. It is concluded that the mimic can be used as an internal control in the diagnosis of L. intracellularis to indicate inhibition of PCR.  相似文献   
35.
The IgG response of goats experimentally infected with RH Toxoplasma gondii has been analysed using an indirect ELISA and Western-blot analysis. Specific IgG antibodies were first detected at 14 days post-inoculation (p.i.), reaching a peak by day 35 p.i. and showing slight fluctuations until the end of the experiment (91 p.i.). Specific IgG showed a reactivity over a whole range of peptides (125-24 kDa approximately), but the highest reactivity was observed against a group of antigens with a molecular weight between 34 and 28 kDa, in particular against a 30 kDa fraction which is considered to represent the major surface protein of T. gondii named p30 or SAG-1.  相似文献   
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Summary Chromosome number of morphogenic and non-morphogenic calli and regenerated plants of barley were determined. Cultures were obtained from two kinds of explants, immature embryos and seedling leaves from three cultivars, Ingrid, Dissa and Golden Promise. Callus chromosome analyses were carried out during a 12 month period in a medium containing 2 mg/l of 2,4-D. Diploid cells were predominant in all cases; although in leaf-derived cultures, retraploid cells (2n=4x=28) showed a tendency to increase as time in culture increased and after more than six months in culture, diploid cells decreased to percentages of almost 70%. Aneuploid cells were generally infrequent in all cases. The source of explant has been more important than the genotype (cultivar) and the type of callus (morphogenic vs. non-morphogenic) in the chromosomal stability of cultures as time increases. From short term cultures, only 1.85% of the regenerated plants were tetraploid, the remaining were diploids. The ability of morphogenic calli to regenerate plants decreased before any significant reduction of diploid cells were observed.  相似文献   
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We have used 19 SSR markers to fingerprint 41 local potato cultivars from 10 locations of Tenerife Island. These varieties represent relicts of the early introductions originating from South America and have been characterised previously morphologically and ecophysiologically. The SSR primers generated a varying degree of polymorphisms. A total of 67 alleles were observed, 12 of them were present in all cultivars. Several accession and group specific alleles were detected. Similarity coefficients were computed from the molecular data and cluster analyses were performed. Generally, cultivar groups with identical or related common names showed the same SSR patterns or clustered closely together. According to the molecular patterns misleading or confounded names were evident for four accessions. The dendrogram clusters were generally in good agreement with previous classifications of the accessions as Solanum tuberosum subsp. andigena, S. tuberosum subsp. tuberosum and Solanum chaucha genotypes. However in four cases the molecular patterns showed discrepancies with previous species assignments suggesting the need for a more detailed and comparative study of these accessions.  相似文献   
39.
The objective of this paper was to study the differences between some prolamin variants coded at the loci Glu-3/Gli-1, Glu-1 and Gli-A2 and their relative roles in durum-wheat quality. F3 lines from four durum wheat crosses (‘Abadia’בMexicali’. ‘Oscar’בArdente’, ‘Oscar × Mexicali’ and ‘Alaga’בC. of Balazote’) were analysed for gliadin and glutenin composition by electrophoresis. Whole-grain-derived samples were analysed for SDS sedimentation (SDSS) value, mixing properties, and contents of protein and vitreousness. The glutenin patterns LMW-2. LMW-2? and LMW-2 (CB) at Glu-B3/Gli-B1 were associated with better gluten quality than were LMW-1 and LMW-2*. The glutenin subunits LMW4 and LMW3 + 15 at Glu-A3/Gli-A1 and HMW-1 showed better mixing properties than LMW7 + 12, LMW5 and the null phenotype. respectively. The HMW glutenin subunits 20 + 8 at Glu-B1 showed a negative association with gluten quality, but the rest of the HMW glutenin subunits and α-gliadins did not show any influence on gluten quality. Correlations between the results of the SDSS test and the mixograph were highly significant, but no correlation was found between these results and protein and vitreousness contents. The results are discussed in relation to the development of durum wheat varieties with improved qualities.  相似文献   
40.
The fatty acid composition and total fatty acid content of seeds from 36 blackcurrant genotypes developed at the Scottish Crop Research Institute were examined. A rapid small-scale procedure, involving homogenization of seeds in toluene followed by sodium methoxide transesterification and gas chromatography, was used. There was considerable variation between genotypes. The gamma-linolenic acid content generally varied from 11 to 19% of the total fatty acids, but three genotypes had higher values of 22-24%, levels previously not reported for blackcurrant seed and similar to those for borage seed. Other nutritionally important fatty acids, stearidonic acid and alpha-linolenic acid, varied from 2 to 4% and 10-19%, respectively. The mean total fatty acid contents ranged from 14 to 23% of the seed, but repeatability was poor. The results are discussed. Blackcurrant seeds are mainly byproducts from juice production, and the study shows the potential for developing blackcurrant genotypes with optimal added value.  相似文献   
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