Sample/analyte screening systems and chromatography

2001 ◽  
Vol 53 (S1) ◽  
pp. S149-S153 ◽  
Author(s):  
M. Valcárcel ◽  
M. Gallego ◽  
S. Cárdenas
Keyword(s):  
Author(s):  
D. A. Bairashewski ◽  
G. Yu. Drobychev ◽  
V. A. Karas ◽  
V. V. Komarov ◽  
M. V. Protsko

2016 ◽  
Vol 83 (6) ◽  
Author(s):  
Tal Argov ◽  
Lev Rabinovich ◽  
Nadejda Sigal ◽  
Anat A. Herskovits

ABSTRACT Construction of Listeria monocytogenes mutants by allelic exchange has been laborious and time-consuming due to lack of proficient selection markers for the final recombination event, that is, a marker conveying substance sensitivity to the bacteria bearing it, enabling the exclusion of merodiploids and selection for plasmid loss. In order to address this issue, we engineered a counterselection marker based on a mutated phenylalanyl-tRNA synthetase gene (pheS*). This mutation renders the phenylalanine-binding site of the enzyme more promiscuous and allows the binding of the toxic p-chloro-phenylalanine analog (p-Cl-phe) as a substrate. When pheS* is introduced into L. monocytogenes and highly expressed under control of a constitutively active promoter, the bacteria become sensitive to p-Cl-phe supplemented in the medium. This enabled us to utilize pheS* as a negative selection marker and generate a novel, efficient suicide vector for allelic exchange in L. monocytogenes. We used this vector to investigate the monocin genomic region in L. monocytogenes strain 10403S by constructing deletion mutants of the region. We have found this region to be active and to cause bacterial lysis upon mitomycin C treatment. The future applications of such an effective counterselection system, which does not require any background genomic alterations, are vast, as it can be modularly used in various selection systems (e.g., genetic screens). We expect this counterselection marker to be a valuable genetic tool in research on L. monocytogenes. IMPORTANCE L. monocytogenes is an opportunistic intracellular pathogen and a widely studied model organism. An efficient counterselection marker is a long-standing need in Listeria research for improving the ability to design and perform various genetic manipulations and screening systems for different purposes. We report the construction and utilization of an efficient suicide vector for allelic exchange which can be conjugated, leaves no marker in the bacterial chromosome, and does not require the use of sometimes leaky inducible promoters. This highly efficient genome editing tool for L. monocytogenes will allow for rapid sequential mutagenesis, introduction of point mutations, and design of screening systems. We anticipate that it will be extensively used by the research community and yield novel insights into the diverse fields studied using this model organism.


2001 ◽  
Vol 16 (supplement) ◽  
pp. 162-163 ◽  
Author(s):  
Toshiyuki KUME ◽  
Tsunehiro HARADA ◽  
Katsuyuki FUKUDA ◽  
Hideshi SHIMADZU

2021 ◽  
Vol 4 (1) ◽  
pp. 71-79
Author(s):  
Borys Igorovych Tymchenko

Nowadays, means of preventive management in various spheres of human life are actively developing. The task of automated screening is to detect hidden problems at an early stage without human intervention, while the cost of responding to them is low. Visual inspection is often used to perform a screening task. Deep artificial neural networks are especially popular in image processing. One of the main problems when working with them is the need for a large amount of well-labeled data for training. In automated screening systems, available neural network approaches have limitations on the reliability of predictions due to the lack of accurately marked training data, as obtaining quality markup from professionals is very expensive, and sometimes not possible in principle. Therefore, there is a contradiction between increasing the requirements for the precision of predictions of neural network models without increasing the time spent on the one hand, and the need to reduce the cost of obtaining the markup of educational data. In this paper, we propose the parametric model of the segmentation dataset, which can be used to generate training data for model selection and benchmarking; and the multi-task learning method for training and inference of deep neural networks for semantic segmentation. Based on the proposed method, we develop a semi-supervised approach for segmentation of salient regions for classification task. The main advantage of the proposed method is that it uses semantically-similar general tasks, that have better labeling than original one, what allows users to reduce the cost of the labeling process. We propose to use classification task as a more general to the problem of semantic segmentation. As semantic segmentation aims to classify each pixel in the input image, classification aims to assign a class to all of the pixels in the input image. We evaluate our methods using the proposed dataset model, observing the Dice score improvement by seventeen percent. Additionally, we evaluate the robustness of the proposed method to different amount of the noise in labels and observe consistent improvement over baseline version.


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