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61.
62.
Andrea Pizzariello Jozef vorc Miroslav Stredansky Stanislav Miertu 《Journal of the science of food and agriculture》1999,79(8):1136-1140
A specific inhibition of 3′,5′‐cyclic phosphodiesterase (CPDE) from bovine heart by methylxanthines was used in combination with a pH electrode to develop a new biosensing method for the detection of caffeine in coffee. The potential response changes of the sensor were proportional to the concentration of caffeine in the range 0–4 mg ml−1. The response time was about 2–4 min. The standard deviation of five measurements of a 0.2 mg ml−1 caffeine solution was ±7.1 µg ml−1. The electrode gave a detection limit of 0.6 mg l−1 caffeine. The concentration of caffeine in espresso coffee was analysed. This model gave excellent correlation between observed and predicted caffeine values. This electrode exhibits advantages such as fast response, short conditioning time and low cost of the instrumentation used. We also expected to be able to perform the detection of caffeine in food and clinical analysis. © 1999 Society of Chemical Industry 相似文献
63.
Asmo Aro-Heinilä Tuomas Lönnberg Dr. Pasi Virta 《Chembiochem : a European journal of chemical biology》2021,22(2):354-358
A highly nucleobase-discriminating metalated nucleoside analogue, 3-fluoro-2-mercuri-6-methylaniline, was incorporated into an oligonucleotide molecular beacon. Fluorescence emission spectra were measured after the addition of four different complementary strands, in which the nucleobase opposite the metalated analogue varies. The fluorescence results showed a clear binding selectivity at room temperature, in the order G>T>C>A. The selectivity is based on the different affinities between the metalated nucleoside analogue and the canonical nucleobases. The synthesized probe is capable of robust discrimination between the two purine as well as the two pyrimidine bases by fluorescence at room temperature, and more sophisticated temperature analysis allows clear separation of every canonical nucleobase. The probe would, hence, be a suitable method for the detection of single nucleotide polymorphisms. 相似文献
64.
Peter Nguyen Kyle Hess Larissa Smulders Dat Le Carolina Briseno Christina M. Chavez Nikolas Nikolaidis 《International journal of molecular sciences》2020,21(24)
Molecular chaperones, particularly the 70-kDa heat shock proteins (Hsp70s), are key orchestrators of the cellular stress response. To perform their critical functions, Hsp70s require the presence of specific co-chaperones, which include nucleotide exchange factors containing the BCL2-associated athanogene (BAG) domain. BAG-1 is one of these proteins that function in a wide range of cellular processes, including apoptosis, protein refolding, and degradation, as well as tumorigenesis. However, the origin of BAG-1 proteins and their evolution between and within species are mostly uncharacterized. This report investigated the macro- and micro-evolution of BAG-1 using orthologous sequences and single nucleotide polymorphisms (SNPs) to elucidate the evolution and understand how natural variation affects the cellular stress response. We first collected and analyzed several BAG-1 sequences across animals, plants, and fungi; mapped intron positions and phases; reconstructed phylogeny; and analyzed protein characteristics. These data indicated that BAG-1 originated before the animals, plants, and fungi split, yet most extant fungal species have lost BAG-1. Furthermore, although BAG-1’s structure has remained relatively conserved, kingdom-specific conserved differences exist at sites of known function, suggesting functional specialization within each kingdom. We then analyzed SNPs from the 1000 genomes database to determine the evolutionary patterns within humans. These analyses revealed that the SNP density is unequally distributed within the BAG1 gene, and the ratio of non-synonymous/synonymous SNPs is significantly higher than 1 in the BAG domain region, which is an indication of positive selection. To further explore this notion, we performed several biochemical assays and found that only one out of five mutations tested altered the major co-chaperone properties of BAG-1. These data collectively suggest that although the co-chaperone functions of BAG-1 are highly conserved and can probably tolerate several radical mutations, BAG-1 might have acquired specialized and potentially unexplored functions during the evolutionary process. 相似文献
65.
B. Fragomeni Y. Masuda H.L. Bradford D.A.L. Lourenco I. Misztal 《Journal of dairy science》2019,102(3):2330-2335
The purpose of this study was to determine whether multi-country genomic evaluation can be accomplished by multiple-trait genomic best linear unbiased predictor (GBLUP) without sharing genotypes of important animals. Phenotypes and genotypes with 40k SNP were simulated for 25,000 animals, each with 4 traits assuming the same genetic variance and 0.8 genetic correlations. The population was split into 4 subpopulations corresponding to 4 countries, one for each trait. Additionally, a prediction population was created from genotyped animals that were not present in the individual countries but were related to each country's population. Genomic estimated breeding values were computed for each country and subsequently converted to SNP effects. Phenotypes were reconstructed for the prediction population based on the SNP effects of a country and the prediction animals' genotypes. The prediction population was used as the basis for the international evaluation, enabling bull comparisons without sharing genotypes and only sharing SNP effects. The computations were such that SNP effects computed within-country or in the prediction population were the same. Genomic estimated breeding values were calculated by single-trait GBLUP for within-country and multiple-trait GBLUP for multi-country predictions. The true accuracy for the prediction population with reconstructed phenotypes was at most 0.02 less than the accuracy with the original data. The differences increased when countries were assumed unequally sized. However, accuracies by multiple-trait GBLUP with the prediction population were always greater than accuracies from any single within-country prediction. Multi-country genomic evaluations by multiple-trait GBLUP are possible without using original genotypes at a cost of lower accuracy compared with explicitly combining countries' data. 相似文献
66.
Bovine leukemia virus (BLV) causes enzootic leucosis in cattle and is classified into 10 genotypes with a worldwide distribution, except for several European countries, Australia, and New Zealand. Although BLV is widespread in Chinese cows with the positive rate of 49.1% at the individual level, very little is known about the BLV genotype in dairy cattle in China. To determine BLV genetic variability in cows in China, 112 BLV-positive samples from 5 cities in China were used for BLV molecular characterization in this study. Phylogenetic analysis using the neighbor-joining method on partial env sequence encoding gp51 obtained from 5 Chinese cities and those available in GenBank (n = 53, representing BLV genotype 1–10) revealed the Chinese strains belonged to genotype 6. Seven unique SNP were identified among Yancheng, Shanghai, and Bengbu strains out of the total 12 SNP identified in Chinese strains. The genotyping coupled with SNP analysis of BLV can serve as a useful molecular epidemiological tool for tracing the source of pathogens. This study highlights the importance of genetic analysis of geographically diverse BLV strains to understand BLV global genetic diversity. 相似文献
67.
68.
In various diseases, there is an increased production of the free radicals needed to carry out certain physiological processes but their excessive amounts can cause oxidative stress and cell damage. Enzymes play a major role in the transformations associated with free radicals. One of them is nitric oxide synthase (NOS), which catalyzes the formation of nitric oxide (NO). This enzyme exists in three forms (NOS1, NOS2, NOS3), each encoded by a different gene. The following work presents the most important information on the NOS isoforms and their role in the human body, including NO synthesis in various tissues and cells, intercellular signaling and activities supporting the immune system and regulating blood vessel functions. The role of NOS in pathological conditions such as obesity, diabetes and heart disease is considered. Attention is also paid to the influence of the polymorphisms of these genes, encoding particular isoforms, on the development of these pathologies and the role of NOS inhibitors in the treatment of patients. 相似文献
69.
The mutations rad3-101 and rad3-102 (formerly rem1-1 and rem1-2) of the essential RAD3 gene of Saccharomyces cerevisiae confer a phenotype of semidominant enhancement of spontaneous mitotic recombination and mutation frequencies, but not extreme sensitivity to ultraviolet (UV) light. These properties differ from the previously published observations of other rad3 mutations, which are very UV-sensitive but do not alter recombination frequencies significantly. We have located the position of DNA sequence changes from wild-type RAD3 to the rad3-101 and rad3-102 mutations and have demonstrated that these sequence changes are necessary and sufficient to confer the (Rem?) mutant phenotype when transferred into otherwise wild-type RAD3 plasmids. The Rem? mutations are not located in the same region. It is possible that the two regions of the gene in which these mutations map define portions of the molecule which are in contact when folded in the native configuration. To begin to test this hypothesis, we have constructed two double mutant alleles, one with rad3-101 and rad3-102, and one with the UV-sensitive rad3-1 mutation and rad3-102. We find that plasmids carrying these double mutant alleles of RAD3 are no longer able to confer a hyper-recombinational phenotype and do not complement the UV-sensitivity of the excision-defective rad3-2 allele. We conclude that the double mutant alleles are non-functional for excision repair, and may be null. We have also constructed new rad3 alleles by oligonucleotide-directed mutagenesis and have tested their effects on spontaneous mutation and mitotic recombination and on UV repair. 相似文献
70.
Cloning and characterization of baker's yeast alpha-glucosidase: over-expression in a yeast strain devoid of vacuolar proteinases 总被引:4,自引:0,他引:4
Two alpha-glucosidase (maltase) genes, designated GLUCPI and GLUCPII, have been cloned from an industrial strain of baker's yeast (Saccharomyces cerevisiae) by complementation of a maltase-negative mutant strain. The different genes were identified according to their alternatively expressed isoenzymes PI and PII in transformants after isoelectric focusing and activity staining in separated cell lysates. The gene encoding alpha-glucosidase PI (GLUCPI), which was not present in laboratory strains of S. carlsbergensis with a defined MAL1, 2, 3, 4 or 6 locus, was sequenced and compared with the recently published MAL6S gene. This comparison revealed single amino acid deviations at three positions in the predicted polypeptide sequence. In addition, the divergent promoter region of GLUCPI differed from MAL6S by a triple repeated 147-bp DNA segment. Maltose induction and glucose repression of alpha-glucosidase PI were not affected by the deletion of the repeated DNA segment. However, the absolute expression of alpha-glucosidase PI increased two- to four-fold. In addition, a two-fold increase in the maltase synthesis occurred when the cloned positive regulator gene MAL2-8ep was on the same plasmid. Furthermore, stability of the alpha-glucosidase in cultures in the stationary growth phase was greatly enhanced using a host strain lacking the proteinases A and B and the carboxypeptidases Y and S. Promoter trimming, MAL2-8cp stimulation and the use of a host strain deficient in four vacuolar proteinases resulted in alpha-glucosidase PI expression of about 13% of the soluble protein. 相似文献