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31.
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Anthocyanin composition in berries of Rhamnus alaternus L., a perennial wild shrub typical of the Mediterranean area, was determined for the first time. The pigments were extracted from the berries with 0.1% HCl in methanol and purified using a C-18 solid-phase cartridge. High-performance liquid chromatography diode array detection-mass spectrometry analysis showed that delphinidin 3-O-rutinoside represented about 62.4% of the total pigments. Other anthocyanins were 3-O-rutinoside derivatives of cyanidin (8.4%), petunidin (15.8%), pelargonidin (4.7%), and peonidin and malvidin (8.7%). The concomitant presence of the six most common anthocyanidins suggested that R. alaternus berries, besides being a good pigment source, could also be a useful tool for anthocyanin identification.  相似文献   
33.
Results from examining shellfish collected from ‘safe’ (2–34Escherichia coli/100 mL) and ‘unsafe’ (> 34E. coli/100 mL) waters for the presence ofYersinia enterocolitica and related bacteria are reported. The recovery of these microorganisms was low in bivalve mollusks destined for food consumption, high in those derived from prohibited areas. No human pathogenic bio-serogroup f Yersinia was found during the entire survey.  相似文献   
34.
In mammalian cells, repair of DNA double-strand breaks (DSBs) by nonhomologous end-joining (NHEJ) is critical for genome stability. Although the end-bridging and ligation steps of NHEJ have been reconstituted in vitro, little is known about the end-processing reactions that occur before ligation. Recently, functionally homologous end-bridging and ligation activities have been identified in prokarya. Consistent with its homology to polymerases and nucleases, we demonstrate that DNA ligase D from Mycobacterium tuberculosis (Mt-Lig) possesses a unique variety of nucleotidyl transferase activities, including gap-filling polymerase, terminal transferase, and primase, and is also a 3' to 5' exonuclease. These enzyme activities allow the Mt-Ku and Mt-Lig proteins to join incompatible DSB ends in vitro, as well as to reconstitute NHEJ in vivo in yeast. These results demonstrate that prokaryotic Ku and ligase form a bona fide NHEJ system that encodes all the recognition, processing, and ligation activities required for DSB repair.  相似文献   
35.
A single genetic alteration, a guanine-to-cytosine transversion, is responsible for the acquisition of malignant properties by K-ras genes of two human tumor cell lines established from carcinomas of the bladder (A1698) and lung (A2182). As a consequence, arginine instead of the normal glycine is incorporated into the K-ras-coded p21 proteins at amino acid position 12. This mutation creates a restriction enzyme polymorphism that can be used to screen human cells for transforming K-ras genes. This approach was used to identify the mutational event responsible for the malignant activation of a K-ras oncogene in a squamous cell lung carcinoma of a 66-year-old man; this point mutation was not present in either the normal bronchial or parenchymal tissue or in the blood lymphocytes. Hence, malignant activation of a ras oncogene appears to be specifically associated with the development of a human neoplasm.  相似文献   
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In eukaryotic cells, double-strand breaks (DSBs) in DNA are generally repaired by the pathway of homologous recombination or by DNA nonhomologous end joining (NHEJ). Both pathways have been highly conserved throughout eukaryotic evolution, but no equivalent NHEJ system has been identified in prokaryotes. The NHEJ pathway requires a DNA end-binding component called Ku. We have identified bacterial Ku homologs and show that these proteins retain the biochemical characteristics of the eukaryotic Ku heterodimer. Furthermore, we show that bacterial Ku specifically recruits DNA ligase to DNA ends and stimulates DNA ligation. Loss of these proteins leads to hypersensitivity to ionizing radiation in Bacillus subtilis. These data provide evidence that many bacteria possess a DNA DSB repair apparatus that shares many features with the NHEJ system of eukarya and suggest that this DNA repair pathway arose before the prokaryotic and eukaryotic lineages diverged.  相似文献   
38.
A simple, rapid, and sensitive spectrophotometric method for routine analysis of saccharin in commercial noncaloric sweeteners is proposed. This method is based on the reaction of saccharin with tetrachloro-p-benzoquinone (p-chloranil) accelerated by hydrogen peroxide and conducted in an ethanol:acetone (4:1) medium, producing a violet-red compound (lambda(max) = 550 nm). Beer's law is obeyed in a concentration range of 2.05 x 10(-4) to 3.00 x 10(-3) M with an excellent correlation coefficient (r = 0.9998). The detection limit was 1.55 x 10(-5) M, and the effect of interferences on the spectrophotometric measurements was evaluated. The proposed procedure was applied successfully to the determination of saccharin in noncaloric sweeteners. Recoveries were within 99.2-104.3% with standard deviations ranging from to 0.5-1.6%. Results of the proposed method compare very favorably with those given by the high-performance liquid chromatography method recommended by the Food and Drug Administration.  相似文献   
39.
Although an antioxidant mechanism has been involved in the beneficial effects of ferulic acid in human diseases, there are few reports on the antioxidant properties of this compound in isolated membranes and intact cells. Here, we evaluated the ability of ferulic acid in inhibiting lipid peroxidation in rat liver microsomal membranes and reactive oxygen species production in NIH-3T3 fibroblasts, induced by both tert-BOOH and AAPH. We also compared its antioxidant efficiency with that of other antioxidants, such as alpha-tocopherol, beta-carotene, and ascorbic acid, added alone or in combination. Ferulic acid acted as a potent antioxidant in our models, being more effective in protecting from tert-BOOH than from AAPH. Moreover, the compound was the most effective among the antioxidants tested. Synergistic interactions were observed when the compound was used in combination with the other antioxidants, suggesting that they can cooperate in preserving physiological integrity of cells exposed to free radicals.  相似文献   
40.
A proteinaceous inhibitor with high activity against trypsin-like serine proteinases was purified from seeds of the tamarind tree (Tamarindus indica) by gel filtration on Shephacryl S-200 followed by a reverse-phase HPLC Vidac C18 TP. The inhibitor, called the tamarind trypsin inhibitor (TTI), showed a Mr of 21.42 kDa by mass spectrometry analysis. TTI was a noncompetitive inhibitor with a Ki value of 1.7 x 10(-9) M. In vitro bioinsecticidal activity against insect digestive enzymes from different orders showed that TTI had remarkable activity against enzymes from coleopteran, Anthonomus grandis (29.6%), Zabrotes subfasciatus (51.6%), Callosobruchus maculatus (86.7%), Rhyzopertha dominica(88.2%), and lepidopteron, Plodia interpuncptella (26.7%), Alabama argillacea (53.8%), and Spodoptera frugiperda (75.5%). Also, digestive enzymes from Diptera, Ceratitis capitata (fruit fly), were inhibited (52.9%). In vivo bioinsecticidal assays toward C. capitata and C. maculatus larvae were developed. The concentration of TTI (w/w) in the artificial seed necessary to cause 50% mortality (LD50) of larvae was 3.6%, and that to reduce mass larvae by 50.0% (ED50) was 3.2%. Furthermore, the mass C. capitata larvae were affected at 53.2% and produced approximately 34% mortality at a level of 4.0% (w/w) of TTI incorporated in artificial diets.  相似文献   
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