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Hafez Hala A. Kamel Maher A. Osman Mohamed Y. Osman Hassan MY. Elblehi Samar S. Mahmoud Shimaa A. 《Molecular and cellular biochemistry》2021,476(5):2233-2249
Molecular and Cellular Biochemistry - Alzheimer’s disease (AD) is a chronic, progressive, multifactorial, and the most common neurodegenerative disease which causes dementia and mental... 相似文献
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Audrey Lenhart Abel Eigege Alphonsus Kal D Pam Emmanuel S Miri George Gerlong J Oneyka Y Sambo J Danboyi B Ibrahim Erica Dahl D Kumbak A Dakul MY Jinadu John Umaru Frank O Richards Tovi Lehmann 《Filaria journal》2007,6(1):1-6
Background
Monitoring and evaluation are essential to the successful implementation of mass drug administration programmes for LF elimination. Monitoring transmission when it is low requires both large numbers of mosquito vectors and sensitive methods for detecting Wuchereria bancrofti infections in them. PCR-based methods are preferred over classical dissections but the best protocol so far achieved detection of one L3 Wuchereria bancrofti larva in a pool of 35–50 Anopheles mosquitoes. It also lacks consistency and remains still a costly tool. Hence we decided to improve upon this to achieve detection in a pool of 100 or more by enhancing the quality of the template DNA. Prior to this we also evaluated three vector sampling methods in the context of numbers for monitoring.Methods
Human landing, pyrethrium spray and light traps catches were conducted concurrently at sites in an LF endemic district in Ghana and the numbers obtained compared. Two DNA extraction methods; Bender buffer and phenol/chloroform purification, and DNAeasy Tissue kit (Quaigen Inc) were used on pools of 25, 50, 75 100 and 150 mosquitoes each seeded with one L3 or its quivalent amount of DNA. Then another set of extracted DNA by the two methods was subjected to Dynal bead purification method (using capture oligonucleotide primers). These were used as template DNA in PCR to amplify W. bancrofti sequences. The best PCR result was then evaluated in the field at five sites by comparing its results (infections per 1000 mosquitoes) with that of dissection of roughly equal samples sizes.Results
The largest numbers of mosquitoes were obtained with the human landing catches at all the sites sampled. Although PCR detection of one L3 in pools of 25, 50 and 75 mosquitoes was consistent irrespective of the extraction method, that of one L3 in 100 was only achieved with the kit-extracted DNA/Dynal bead purification method. Infections were found at only two sites by both dissection and pool-screening being 14.3 and 19 versus 13.4 and 20.1 per 1000 Anopheles mosquitoes respectively, which were not statistically significantDiscussion and conclusion
HLC still remains the best option for sampling for the large numbers of mosquitoes required for monitoring transmission during MDA programmes, when vector population densities are high and classical indices of transmission are required. One – in – 100 detection is an improvement on previous PCR pool-screening methods, which in our opinion was a result of the introduction of the extra step of parasite DNA capture using Dynal/beads. As pool sizes increase the insects DNA will swamp parasite DNA making the latter less available for an efficient PCR, therefore we propose either additional steps of parasite DNA capture or real-time PCR to improve further the pool screening method. The study also attests also to the applicability of Katholi et al's algorithm developed for determining onchocerciasis prevalence in LF studies. 相似文献13.
Background
The goal of DNA barcoding is to develop a species-specific sequence library for all eukaryotes. A 650 bp fragment of the cytochrome c oxidase 1 (CO1) gene has been used successfully for species-level identification in several animal groups. It may be difficult in practice, however, to retrieve a 650 bp fragment from archival specimens, (because of DNA degradation) or from environmental samples (where universal primers are needed).Results
We used a bioinformatics analysis using all CO1 barcode sequences from GenBank and calculated the probability of having species-specific barcodes for varied size fragments. This analysis established the potential of much smaller fragments, mini-barcodes, for identifying unknown specimens. We then developed a universal primer set for the amplification of mini-barcodes. We further successfully tested the utility of this primer set on a comprehensive set of taxa from all major eukaryotic groups as well as archival specimens.Conclusion
In this study we address the important issue of minimum amount of sequence information required for identifying species in DNA barcoding. We establish a novel approach based on a much shorter barcode sequence and demonstrate its effectiveness in archival specimens. This approach will significantly broaden the application of DNA barcoding in biodiversity studies. 相似文献14.
RÉMY GOURVENNEC 《Lethaia: An International Journal of Palaeontology and Stratigraphy》1989,22(4):405-411
Capillae are a major feature of the fine ornamentation in spiriferid brachiopods. Within the lower Devonian spiriferids, studied in this paper, two categories of organization can be distinguished: strictly radial orientation of the microstructures, which leads to the eospiriferid-type of organization, and pseudoradial orientation, which provides a delthyridid-type. Apart from this distinction, it can now be shown that the mode of development is quite different in each case. Thus, and allowing for differences in size, the capillae of eospiriferids are quite similar to the costae of costate spiriferids. On the other hand, the origin of the capillae is more complex in the delthyridids and as yet not fully known. Other factors (such as the rate of growth, the divergence angle of the capillae, etc.) have effects on the growth mechanisms and contribute to the existence of a large variety of microornaments. Furthermore. available data indicate that the divergence of the capillae is closely related to the paleogeographical distribution of the populations involved. If this observation proves valuable for the whole stock of spiriferids ( s.l. ), it opens new ground for paleozoogeographical investigations. □ Spiriferid brachiopods, eospiriferids, delthyridids. microornament, Devonian, paleogeography. 相似文献
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The glucose transport system of the hyperthermophilic anaerobic bacterium Thermotoga neapolitana 总被引:1,自引:1,他引:0 下载免费PDF全文
The glucose transport system of the extremely thermophilic anaerobic bacterium Thermotoga neapolitana was studied with the nonmetabolizable glucose analog 2-deoxy-D-glucose (2-DOG). T. neapolitana accumulated 2-DOG against a concentration gradient in an intracellular free sugar pool that was exchangeable with external source of energy, such as pyruvate, and was inhibited by arsenate and gramicidin D. There was no phosphoenolpyruvate-dependent phosphorylation of glucose, 2-DOG, or fructose by cell extracts or toluene-treated cells, indicating the absence of a phosphoenolpyruvate:sugar phosphotransferase system. These data indicate that D-glucose is taken up by T. neapolitana via an active transport system that is energized by an ion gradient generated by ATP, derived from substrate-level phosphorylation. 相似文献
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Liquid chromatographic determination of isoniazid, pyrazinamide and rifampicin from pharmaceutical preparations and blood 总被引:2,自引:0,他引:2
Khuhawar MY Rind FM 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2002,766(2):357-363
Isoniazid (IN), pyrazinamide (Pz) and rifampicin (Rf) are separated on YMC-ODS column. IN was derivatized with 2-fluorene-carboxaldehyde (FA). The separation was achieved using ethanol-chloroform-acetonitrile water by isocratic elution and detected at 337 nm. The detection limits were 0.11 ng, 0.2 ng and 13 ng/injection (5 microl) for IN, Pz and Rf, respectively. The method of analysis was applied to the pharmaceutical preparations and in the blood samples of the patients suffering from tuberculosis after undergoing chemotherapy with IN, Pz and Rf. The amounts quantitated in blood showed 0.97 to 1.58 microg/ml IN, 3.44 to 4.09 microg/ml Pz and 1.98 to 3.5 microg/ml Rf with coefficient of variations 0.8-1.8%, 0.9-1.3% and 0.8-2.1%, respectively. 相似文献
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A novel approach to the design of sensitive fluorescent probes for nucleic acids detection is proposed. Suitable modifications of tri- and pentamethine cyanine dyes in the polymethine chain and/or in the heterocyclic residues can result in a significant decrease in unbound dye fluorescence intensity and an increase in dye emission intensity in the presence of DNA compared to the unsubstituted dye. The sharp enhancement in the fluorescence intensity upon dye interaction with double-stranded DNA permits the application of the modified tri- and pentamethine dyes as fluorescent probes in double-stranded DNA detection in homogeneous assays. 相似文献