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Detection Protocol (detection + protocol)
Selected AbstractsSelective Electroanalytical Assay for Cysteine at a Boron Doped Diamond ElectrodeELECTROANALYSIS, Issue 16 2004Olga Nekrassova Abstract The electrochemical oxidation of the cysteine-quinone adduct has been examined as a means of providing an electroanalytical cysteine specific detection protocol. The appliance of square-wave voltammetry allowed 0.5,,M as a limit of detection. The effects of various biologically relevant interferences including other thiols were studied and found to present no change in the voltammetric profile. The practical applicability and efficiency of the methodology was probed through the determination of cysteine concentration in growth tissue medium. [source] Performance of collaborative codes in CSMA/CD environmentEUROPEAN TRANSACTIONS ON TELECOMMUNICATIONS, Issue 5 2006F. Gebali A new medium access control scheme is proposed for implementing collaborative codes in a system using carrier sense multiple access with collision detection protocol (CC-CSMA/CD). We also propose a new backoff algorithm which is simple to implement and to analyse. A discrete-time Markov chain analytical model is developed for CC-CSMA/CD. The resulting model describes the regular CSMA/CD as a special case. Protocol performance measures were studied such as throughput, packet acceptance probability, average packet delay and channel utilisation. It is found that CC-CSMA/CD offers improvements over a system that uses CSMA/CD in terms of throughput, packet acceptance probability, delay and channel utilisation. Copyright © 2006 AEIT. [source] Effective detection of corrected dystrophin loci in mdx mouse myogenic precursors,HUMAN MUTATION, Issue 8 2007Marian Todaro Abstract Targeted corrective gene conversion (TCGC) holds much promise as a future therapy for many hereditary diseases in humans. Mutation correction frequencies varying between 0.0001% and 40% have been reported using chimeraplasty, oligoplasty, triplex-forming oligonucleotides, and small corrective PCR amplicons (CPA). However, PCR technologies used to detect correction events risk either falsely indicating or greatly exaggerating the presence of corrected loci. This is a problem that is considerably exacerbated by attempted improvement of the TCGC system using high corrective nucleic acid (CNA) to nuclear ratios. Small fragment homologous replacement (SFHR)-mediated correction of the exon 23 dystrophin (DMD) gene mutation in the mdx mouse model of DMD has been used in this study to evaluate the effect of increasing CPA amounts. In these experiments, we detected extremely high levels of apparently corrected loci and determined that at higher CNA to nuclear ratios the extent of locus correction was highly exaggerated by residual CNA species in the nucleic acids extracted from the treated cells. This study describes a generic locus-specific detection protocol designed to eradicate residual CNA species and avoid the artifactual or exaggerated detection of gene correction. Hum Mutat 28(8), 816,823, 2007. © 2007 Wiley-Liss, Inc. [source] Field studies on the environmental fate of the Cry1Ab Bt-toxin produced by transgenic maize (MON810) and its effect on bacterial communities in the maize rhizosphereMOLECULAR ECOLOGY, Issue 8 2005SUSANNE BAUMGARTE Abstract Field studies were done to assess how much of the transgenic, insecticidal protein, Cry1Ab, encoded by a truncated cry1Ab gene from Bacillus thuringiensis (Bt), was released from Bt-maize MON810 into soil and whether bacterial communities inhabiting the rhizosphere of MON810 maize were different from those of the rhizosphere of nontransgenic maize cultivars. Bacterial community structure was investigated by SSCP (single-strand conformation polymorphism) of PCR-amplified 16S rRNA genes from community DNA. Using an improved extraction and detection protocol based on a commercially available ELISA, it was possible to detect Cry1Ab protein extracted from soils to a threshold concentration of 0.07 ng/g soil. From 100 ng of purified Cry1Ab protein added per gram of soil, only an average of 37% was extractable. At both field sites investigated, the amount of Cry1Ab protein in bulk soil of MON810 field plots was always lower than in the rhizosphere, the latter ranging from 0.1 to 10 ng/g soil. Immunoreactive Cry1Ab protein was also detected at 0.21 ng/g bulk soil 7 months after harvesting, i.e. in April of the following year. At this time, however, higher values were found in residues of leaves (21 ng/g) and of roots (183 ng/g), the latter corresponding to 12% of the Cry1Ab protein present in intact roots. A sampling 2 months later indicated further degradation of the protein. Despite the detection of Cry1Ab protein in the rhizosphere of MON810 maize, the bacterial community structure was less affected by the Cry1Ab protein than by other environmental factors, i.e. the age of the plants or field heterogeneities. The persistence of Cry1Ab protein emphasizes the importance of considering post-harvest effects on nontarget organisms. [source] A qualitative proteome investigation of the sediment portion of human urine: Implications in the biomarker discovery processPROTEOMICS - CLINICAL APPLICATIONS, Issue 1 2009Diane Mataija-Botelho Abstract Inherent to the biomarker discovery process is a comparative analysis of physiological states. It is therefore critical that the proteome detection protocol does not bias the analysis. With urine, the sediment portion, obtained upon thawing frozen urine, is routinely discarded prior to proteome analysis. However, our results demonstrate that such a practice inadvertently induces bias, having significant implications in the biomarker discovery process. We present the first proteome investigation of human urinary sediments, identifying 60 proteins in this phase by MS. Many sediment proteins were also detected in the urinary supernatant, indicating that several proteins partition between the two phases. This partitioning is dependant on the pH of the sample, as well as the degree of sample agitation. As a consequence of discarding the sediment portion of urine, the concentration of potential candidate biomarkers in the supernatant phase will be altered or, in other instances, may be completely removed from the sample. To minimize this, the pH of all samples should first be normalized, and the samples vigorously vortexed prior to discarding the sediments. For more comprehensive biomarker investigations, we suggest that urinary sediments be analyzed along with the supernatant proteins. [source] Detecting North American signal crayfish (Pacifastacus leniusculus) in rifflesAQUATIC CONSERVATION: MARINE AND FRESHWATER ECOSYSTEMS, Issue 5 2010Z. F. Gladman Abstract 1.The spread of the invasive signal crayfish (Pacifastacus leniusculus) outside its natural range is of widespread concern due to the threats posed to native biodiversity. To date, there is no standard protocol for determining signal crayfish presence or absence in a watercourse. 2.For the purposes of this investigation, the crayfish detection ability of active sampling methods , hand-netting, electrofishing (one, two and three runs), kick sampling and Surber sampling , was tested at 30 sites along the River Clyde, southern central Scotland. 3.No single technique was successful in detecting crayfish in 100% of the sites known to contain crayfish and so the application of combinations of techniques was considered. The combination of techniques that resulted in a 100% detection rate was electrofishing (three runs) together with kick sampling. These results suggest that three-run electrofishing and kick sampling are the best candidates for incorporation into a crayfish detection protocol. 4.The mean time taken to apply electrofishing (three runs) was significantly greater than the mean time to apply kick sampling. Given the lower effort required for its application, kick sampling is recommended as the preliminary technique: if kick sampling yields a negative result, the application of electrofishing will decrease the chance of recording a false negative presence. If both kick sampling and electrofishing fail to detect crayfish, trapping may further decrease the risk of a false negative result. 5.These findings have assisted in the development of a crayfish detection protocol, which will be applied across Scotland to determine the current distribution of signal crayfish. Copyright © 2010 John Wiley & Sons, Ltd. [source] HCV-RNA In Sural Nerve From Hcv Infected Patients With Peripheral NeuropathyJOURNAL OF THE PERIPHERAL NERVOUS SYSTEM, Issue 1 2001L De Martino Objective: Evaluation of hepatitis C virus (HCV) by reverse transcription-polymerase chain reaction (RT-PCR) in peripheral nerve tissues from HCV infected patients with peripheral neuropathy. METHODS: RT-PCR was performed on homogenates of nerve biopsies from 17 consecutive HCV-positive patients with peripheral neuropathy, with or without mixed cryoglobulinemia, hospitalised from 1996 to 2000. Sural nerve specimens were frozen in iso-pentane pre-cooled in liquid nitrogen and stored at ,80°C until use. RNA was extracted from ten 7-,m thick cryostatic sections or from a nerve trunk specimen of about 3 mm length, collected from each biopsy. Three different protocols of RNA extraction were tested (1,3). Complementary DNAs (cDNAs) were obtained without or with RNasin (Promega, Madison, WI) addition in the reaction mixture to inhibit residual RNase activity. Two sets of commercially available PCR primers for the outer and the nested reaction were used. PCR products were analysed by agarose gel electrophoresis and ethidium bromide staining. Serum samples and liver specimens from proven HCV positive patients served as positive controls, whereas sera from healthy subjects were negative controls. RESULTS: Sufficient amount of RNA could be obtained either by cryostatic sections or by in toto nerve specimens. Extraction by Trizol (Gibco-BRL) allowed the best concentration and purity of RNA as assessed by biophotometry. The presence of RNasin didn't improve the cDNA synthesis. The resulting amplification product of the nested PCR was 187 bp long. We have always observed this product in our positive controls and never in the negative. Six samples from patients either with or without cryoglobulinemia resulted positive; 7 were negative. Four samples gave variable results. CONCLUSIONS: While 40% of the nerves in our series were undoubtedly HCV positive, the cause(s) of negative and variable results in the remaining samples is likely more complex than variations in the detection protocols and deserve further investigations. REFERENCES: 1) Chomczynski P, Sacchi N (1987). Anal Biochem 162:156. 2) Marquardt O et al. (1996). Med Microbiol Lett 5:55. 3) Chomczynski P (1993). Bio/Techniques 15:532. [source] | ||||||||||