Somatic Mutation Detection in Human Biomonitoring

Olsen, L. S.; Nielsen, Lise Rud; Nexø, Bjørn A.; Wassermann, Karsten

doi:10.1111/j.1600-0773.1996.tb00220.x

Cited by 15 publications

(4 citation statements)

References 73 publications

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“…[1–5] which have been the gold standard for many years but these assays are costly and can take weeks to conduct. Gene mutation assays such as HPRT [1,6,7] and Glycophorin A [1,6,8,9] have also been used, although these are not usually as accurate as cytogenetics [1] and only work on individuals of the appropriate genotype or gender. Electron paramagnetic resonance has also been used successfully [9].…”

Section: Introductionmentioning

confidence: 99%

Gene Expression-Based Dosimetry by Dose and Time in Mice Following Acute Radiation Exposure

et al. 2013

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Rapid and reliable methods for performing biological dosimetry are of paramount importance in the event of a large-scale nuclear event. Traditional dosimetry approaches lack the requisite rapid assessment capability, ease of use, portability and low cost, which are factors needed for triaging a large number of victims. Here we describe the results of experiments in which mice were acutely exposed to 60Co gamma rays at doses of 0 (control) to 10 Gy. Blood was obtained from irradiated mice 0.5, 1, 2, 3, 5, and 7 days after exposure. mRNA expression levels of 106 selected genes were obtained by reverse-transcription real time PCR. Stepwise regression of dose received against individual gene transcript expression levels provided optimal dosimetry at each time point. The results indicate that only 4–7 different gene transcripts are needed to explain ≥ 0.69 of the variance (R2), and that receiver-operator characteristics, a measure of sensitivity and specificity, of ≥ 0.93 for these statistical models were achieved at each time point. These models provide an excellent description of the relationship between the actual and predicted doses up to 6 Gy. At doses of 8 and 10 Gy there appears to be saturation of the radiation-response signals with a corresponding diminution of accuracy. These results suggest that similar analyses in humans may be advantageous for use in a field-portable device designed to assess exposures in mass casualty situations.

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Section: Introductionmentioning

confidence: 99%

Gene Expression-Based Dosimetry by Dose and Time in Mice Following Acute Radiation Exposure

et al. 2013

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“…A simple, sensitive, speci®c and ecient method for identifying point mutations in nucleic acids is needed. Several methods have been developed and meet these requirements to varying degrees (Grompe, 1993;Olsen et al, 1996). Manual dideoxy termination sequencing (Sanger et al, 1977), the accepted standard for identi®cation of alterations in nucleic acid sequences, is both time-and labor-intensive.…”

Section: Introductionmentioning

confidence: 99%

Detection of mutations by automated fluorescence/RNA-based dideoxy fingerprinting (ARddF)

Martincic

Koury

Gale³

et al. 1999

Oncogene

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Dideoxy ®ngerprinting (ddF) is a hybrid technique which combines aspects of single strand conformational polymorphism (SSCP) and dideoxy sequencing to detect the presence of single base changes in a de®ned fragment of nucleic acid. ddF is no more technically demanding than SSCP, yet it is more sensitive in detecting point mutations. We describe here the adaptation of conventional ddF to an automated sequencing system usinḡ uorescent Cy5 labeled primers. We show that automated RNA-based ddF (ARddF) has several advantages over conventional radioisotope-based ddF, including: (1) analysis of larger nucleic acid fragments (up to 10 3 bp), due to the ability to continuously analyse and compile sequencing information; (2) greater reliability for distinguishing mutant sequences from wild type sequences (particularly when the mutation leads to gain or loss of a dideoxy termination segment); (3) the use of¯uorescent labeled primers, making ARddF less hazardous than methods requiring radionucleotides. The use of ARddF in conjunction with new methods for isolating RNA from a small number of cells facilitates mutational analysis of small tissue biopsies and other limited samples, and will allow more widespread application of mutational screening in the setting of clinical diagnostic laboratories.

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“…Recombinant enzymes exhibiting better activity and/or stability have been obtained as a result of mutagenesis [16] or gene fusion [86,67]. However cleavage of protein fusions to generate free protein remains the major disadvantage of protein fusion technologies.…”

Section: Construction Of Bifunctional Xylanase By Gene Fusionmentioning

confidence: 99%

Bifunctional xylanases and their potential use in biotechnology

Khandeparker

Numan

2008

J Ind Microbiol Biotechnol

112

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Plant cell walls are comprised of cellulose, hemicellulose and other polymers that are intertwined. This complex structure acts as a barrier to degradation by single enzyme. Thus, a cocktail consisting of bi and multifunctional xylanases and xylan debranching enzymes is most desired combination for the efficient utilization of these complex materials. Xylanases have prospective applications in the food, animal feed, and paper and pulp industries. Furthermore, in order to enhance feed nutrient digestibility and to improve wheat flour quality xylanase along with other glycohydrolases are often used. For these applications, a bifunctional enzyme is undoubtedly much more valuable as compared to monofunctional enzyme. The natural diversity of enzymes provides some candidates with evolved bifunctional activity. Nevertheless most resulted from the in vitro fusion of individual enzymes. Here we present bifunctional xylanases, their evolution, occurrence, molecular biology and potential uses in biotechnology.

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Somatic Mutation Detection in Human Biomonitoring

Cited by 15 publications

References 73 publications

Gene Expression-Based Dosimetry by Dose and Time in Mice Following Acute Radiation Exposure

Gene Expression-Based Dosimetry by Dose and Time in Mice Following Acute Radiation Exposure

Detection of mutations by automated fluorescence/RNA-based dideoxy fingerprinting (ARddF)

Bifunctional xylanases and their potential use in biotechnology

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