scholarly journals Gamma-H2AX biodosimetry for use in large scale radiation incidents: comparison of a rapid lyse/fix protocol with a routine method

2013 ◽  
Author(s):  
Jayne Moquet ◽  
Stephen G R Barnard ◽  
Kai Rothkamm
Keyword(s):  
Large Scale ◽  
Processing Time ◽  
Clinical Symptoms ◽  
Gradient Centrifugation ◽  
Routine Method ◽  
Sample Processing ◽  
Ficoll Density Gradient Centrifugation ◽  
One Step ◽  
The Cost ◽  
Ficoll Density Gradient

Following a radiation incident, preliminary dose estimates made by γ-H2AX foci analysis can supplement the early triage of casualties based on clinical symptoms. Sample processing time is important when many individuals need to be rapidly assessed. A protocol was therefore developed for high sample throughput that requires less than 0.1 ml blood, thus enabling finger prick sampling. The technique combines red blood cell lysis and leukocyte fixation in one step on a 96 well plate, in contrast to the routine protocol, where lymphocytes are separated by Ficoll density gradient centrifugation with subsequent washing and fixation steps. The rapid lyse/fix method reduced the estimated sample processing time for 96 samples to about 4 h compared to 15 h using the routine protocol. However, scoring 20 cells in 96 samples prepared by the rapid protocol took longer than for the routine method (3.1 versus 1.5 h at zero dose; 7.0 versus 6.1 h for irradiated samples). Similar foci yields were scored for both protocols and reliable dose estimates were obtained for coded samples, with mean absolute differences from the actual doses of 0.26 and 0.27 Gy for the routine and lyse/fix method, respectively. The lyse/fix protocol can therefore facilitate high throughput processing for γ-H2AX biodosimetry for use in large scale radiation incidents, at the cost of somewhat longer foci scoring times.

scholarly journals Gamma-H2AX biodosimetry for use in large scale radiation incidents: comparison of a rapid lyse/fix protocol with a routine method

2013 ◽  
Author(s):  
Jayne Moquet ◽  
Stephen G R Barnard ◽  
Kai Rothkamm
Keyword(s):  
Large Scale ◽  
Processing Time ◽  
Clinical Symptoms ◽  
Gradient Centrifugation ◽  
Routine Method ◽  
Sample Processing ◽  
Ficoll Density Gradient Centrifugation ◽  
One Step ◽  
The Cost ◽  
Ficoll Density Gradient

Following a radiation incident, preliminary dose estimates made by γ-H2AX foci analysis can supplement the early triage of casualties based on clinical symptoms. Sample processing time is important when many individuals need to be rapidly assessed. A protocol was therefore developed for high sample throughput that requires less than 0.1 ml blood, thus enabling finger prick sampling. The technique combines red blood cell lysis and leukocyte fixation in one step on a 96 well plate, in contrast to the routine protocol, where lymphocytes are separated by Ficoll density gradient centrifugation with subsequent washing and fixation steps. The rapid lyse/fix method reduced the estimated sample processing time for 96 samples to about 4 h compared to 15 h using the routine protocol. However, scoring 20 cells in 96 samples prepared by the rapid protocol took longer than for the routine method (3.1 versus 1.5 h at zero dose; 7.0 versus 6.1 h for irradiated samples). Similar foci yields were scored for both protocols and reliable dose estimates were obtained for coded samples, with mean absolute differences from the actual doses of 0.26 and 0.27 Gy for the routine and lyse/fix method, respectively. The lyse/fix protocol can therefore facilitate high throughput processing for γ-H2AX biodosimetry for use in large scale radiation incidents, at the cost of somewhat longer foci scoring times.

scholarly journals Function of miR-24 and miR-27 in Pediatric Patients With Idiopathic Nephrotic Syndrome

2021 ◽  
Author(s):  
Fen-fen Ni ◽  
Guang-lei Liu ◽  
shi-lei Jia ◽  
Ran-ran Chen ◽  
Li-bing Liu ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Nephrotic Syndrome ◽  
Idiopathic Nephrotic Syndrome ◽  
Gradient Centrifugation ◽  
Th2 Cells ◽  
Rt Pcr ◽  
Critical Function ◽  
Significant Difference ◽  
Ficoll Density Gradient Centrifugation ◽  
Ficoll Density Gradient

Abstract Background: The specific etiology and mechanism of idiopathic nephrotic syndrome (INS) in children remain unclear, so we investigated the pathogenesis of INS by measuring the effects two specific miRNAs on Th2 cells in children with this disease. Methods: Flow cytometry was used to measure the levels of Th2 cells and a cytometric bead array was used to measure the levels of IgE, interleukin (IL) -4, and IL-13. RT-PCR was used to measure the levels of miR-24 and miR-27 in CD4+TCD25− cells. PBMCs were isolated using Ficoll density gradient centrifugation, and transfected with different mimic or inhibitor miRNAs. RT-PCR was used to measure the expression of different RNAs, and flow cytometry was used to determine the percentages of Th2 cells. Results: Children with active INS had higher percentages of Th2 cells than healthy controls (P<0.05), but there was no significant difference for children in remission. The plasma levels of IgE, IL-4, and IL-13 were significantly increased in children with active INS (P<0.05). miR-24 and miR-27 were at lower levels in children with active non-atopic INS (P<0.05). Transfection experiments indicated that upregulation of each miRNA decreased the percentage of Th2 cells and the level of IL-4 (P<0.05), and down-regulation of each miRNA had the opposite effects (P<0.05). Conclusion: Children with active INS, with or without atopy, had higher levels of IgE, possibly related to their higher levels of IL-13 and IL-4 due to drift toward Th2 cells. miR-24 and miR-27 suppress the expression of Th2 cells and have a critical function in Th2 expression in INS.

scholarly journals CHOSEN ASPECTS OF THE PRODUCTION OF THE BASIC MAP USING UAV IMAGERY

2016 ◽  
Vol XLI-B1 ◽  
pp. 873-877 ◽  
Author(s):  
M. Kedzierski ◽  
A. Fryskowska ◽  
D. Wierzbicki ◽  
P. Nerc
Keyword(s):  
Large Scale ◽  
Processing Time ◽  
Reference Data ◽  
Image Data ◽  
Flight Time ◽  
Digital Cameras ◽  
Geometric Accuracy ◽  
Aerial Photos ◽  
Low Altitude ◽  
The Cost

For several years there has been an increasing interest in the use of unmanned aerial vehicles in acquiring image data from a low altitude. Considering the cost-effectiveness of the flight time of UAVs vs. conventional airplanes, the use of the former is advantageous when generating large scale accurate ortophotos. Through the development of UAV imagery, we can update large-scale basic maps. These maps are cartographic products which are used for registration, economic, and strategic planning. On the basis of these maps other cartographic maps are produced, for example maps used building planning. The article presents an assessesment of the usefulness of orthophotos based on UAV imagery to upgrade the basic map. In the research a compact, non-metric camera, mounted on a fixed wing powered by an electric motor was used. The tested area covered flat, agricultural and woodland terrains. The processing and analysis of orthorectification were carried out with the INPHO UASMaster programme. Due to the effect of UAV instability on low-altitude imagery, the use of non-metric digital cameras and the low-accuracy GPS-INS sensors, the geometry of images is visibly lower were compared to conventional digital aerial photos (large values of phi and kappa angles). Therefore, typically, low-altitude images require large along- and across-track direction overlap – usually above 70 %. As a result of the research orthoimages were obtained with a resolution of 0.06 meters and a horizontal accuracy of 0.10m. Digitized basic maps were used as the reference data. The accuracy of orthoimages vs. basic maps was estimated based on the study and on the available reference sources. As a result, it was found that the geometric accuracy and interpretative advantages of the final orthoimages allow the updating of basic maps. It is estimated that such an update of basic maps based on UAV imagery reduces processing time by approx. 40%.

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