scholarly journals A Cytopathic Effect-Based Tissue Culture Method for HCoV-OC43 Titration Using TMPRSS2-Expressing VeroE6 Cells

mSphere ◽  
2021 ◽  
Vol 6 (3) ◽  
Author(s):  
Ryohei Hirose ◽  
Naoto Watanabe ◽  
Risa Bandou ◽  
Takuma Yoshida ◽  
Tomo Daidoji ◽  
...  

ABSTRACT Human coronavirus (HCoV)-OC43 rarely shows a cytopathic effect (CPE) after infection of various cell lines, and the indirect immunoperoxidase assay (IPA), a relatively complex procedure, has long been used as an alternative assay. Because HCoV-OC43 uses cell-surface transmembrane protease serine 2 (TMPRSS2) for cell entry, VeroE6 cells expressing TMPRSS2 may show a clear CPE after HCoV-OC43 infection. The aim of this study was to construct a 50% tissue culture infectious dose (TCID50) assay for HCoV-OC43 based on CPE evaluation using VeroE6/TMPRSS2 cells. VeroE6/TMPRSS2 cells showed clear CPEs 3 to 4 days after low-titer HCoV-OC43 infection. Evaluation of viral kinetics indicated that the viral titer in the culture supernatant of VeroE6/TMPRSS2 cells in the early stages of infection was higher than that of other cells. In comparison, between the CPE-based and the IPA-based (i.e., the reference titer) methods, the titer measured with CPE evaluation 4 to 5 days after infection using VeroE6/TMPRSS2 cells showed a much smaller difference from the reference titer than that measured using other cells. Thus, the TCID50 assay using CPE evaluation with VeroE6/TMPRSS2 cells provides the correct titer value and will greatly contribute to future research on HCoV-OC43. IMPORTANCE HCoV-OC43 rarely shows a cytopathic effect (CPE) in infected cell lines, and thus the plaque and TCID50 assays by CPE observation are not applicable for titration; the indirect immunoperoxidase assay (IPA) is used instead. However, the IPA is relatively complex, time-consuming, costly, and not suitable for simultaneous titration of many samples. We developed a TCID50 assay using CPE evaluation with TMPRSS2-expressing VeroE6/TMPRSS2 cells that provides the same accuracy as the conventional IPA-based viral titration and does not require any staining procedures using antibodies or substrates. This titration method will greatly contribute to future research on HCoV-OC43 by allowing simple, low-cost, and accurate titration of this virus.

1978 ◽  
Vol 26 (11) ◽  
pp. 934-947 ◽  
Author(s):  
E Karsenti ◽  
B Guilbert ◽  
M Bornens ◽  
S Avrameas ◽  
R Whalen ◽  
...  

This paper reports on the preparation of immunsera against tubulin and actin, and the purification of anti-tubulin and anti-actin antibodies on immunoadsorbent columns. These purified antibodies were used in an indirect immunoperoxidase assay to visualize microtubules and microfilaments in various cell lines. The specificity of antibodies and the methods of cell fixation required are discussed, as well as some aspects of microtubule and microfilament organization, as visualized by this technique.


2018 ◽  
Vol 32 (2) ◽  
pp. 103-119
Author(s):  
Colleen M. Boland ◽  
Chris E. Hogan ◽  
Marilyn F. Johnson

SYNOPSIS Mandatory existence disclosure rules require an organization to disclose a policy's existence, but not its content. We examine policy adoption frequencies in the year immediately after the IRS required mandatory existence disclosure by nonprofits of various governance policies. We also examine adoption frequencies in the year of the subsequent change from mandatory existence disclosure to a disclose-and-explain regime that required supplemental disclosures about the content and implementation of conflict of interest policies. Our results suggest that in areas where there is unclear regulatory authority, mandatory existence disclosure is an effective and low cost regulatory device for encouraging the adoption of policies desired by regulators, provided those policies are cost-effective for regulated firms to implement. In addition, we find that disclose-and-explain regulatory regimes provide stronger incentives for policy adoption than do mandatory existence disclosure regimes and also discourage “check the box” behavior. Future research should examine the impact of mandatory existence disclosure rules in the year that the regulation is implemented. Data Availability: Data are available from sources cited in the text.


Electronics ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 517
Author(s):  
Seong-heum Kim ◽  
Youngbae Hwang

Owing to recent advancements in deep learning methods and relevant databases, it is becoming increasingly easier to recognize 3D objects using only RGB images from single viewpoints. This study investigates the major breakthroughs and current progress in deep learning-based monocular 3D object detection. For relatively low-cost data acquisition systems without depth sensors or cameras at multiple viewpoints, we first consider existing databases with 2D RGB photos and their relevant attributes. Based on this simple sensor modality for practical applications, deep learning-based monocular 3D object detection methods that overcome significant research challenges are categorized and summarized. We present the key concepts and detailed descriptions of representative single-stage and multiple-stage detection solutions. In addition, we discuss the effectiveness of the detection models on their baseline benchmarks. Finally, we explore several directions for future research on monocular 3D object detection.


Rare Metals ◽  
2021 ◽  
Author(s):  
Jia-Xing Song ◽  
Xin-Xing Yin ◽  
Zai-Fang Li ◽  
Yao-Wen Li

Abstract As a promising photovoltaic technology, perovskite solar cells (pero-SCs) have developed rapidly over the past few years and the highest power conversion efficiency is beyond 25%. Nowadays, the planar structure is universally popular in pero-SCs due to the simple processing technology and low-temperature preparation. Electron transport layer (ETL) is verified to play a vital role in the device performance of planar pero-SCs. Particularly, the metal oxide (MO) ETL with low-cost, superb versatility, and excellent optoelectronic properties has been widely studied. This review mainly focuses on recent developments in the use of low-temperature-processed MO ETLs for planar pero-SCs. The optical and electronic properties of widely used MO materials of TiO2, ZnO, and SnO2, as well as the optimizations of these MO ETLs are briefly introduced. The commonly used methods for depositing MO ETLs are also discussed. Then, the applications of different MO ETLs on pero-SCs are reviewed. Finally, the challenge and future research of MO-based ETLs toward practical application of efficient planar pero-SCs are proposed. Graphical abstract


Sensors ◽  
2019 ◽  
Vol 19 (22) ◽  
pp. 4916 ◽  
Author(s):  
Qiaoyun Wu ◽  
Yunzhe Zhang ◽  
Qian Yang ◽  
Ning Yuan ◽  
Wei Zhang

The vital importance of rapid and accurate detection of food borne pathogens has driven the development of biosensor to prevent food borne illness outbreaks. Electrochemical DNA biosensors offer such merits as rapid response, high sensitivity, low cost, and ease of use. This review covers the following three aspects: food borne pathogens and conventional detection methods, the design and fabrication of electrochemical DNA biosensors and several techniques for improving sensitivity of biosensors. We highlight the main bioreceptors and immobilizing methods on sensing interface, electrochemical techniques, electrochemical indicators, nanotechnology, and nucleic acid-based amplification. Finally, in view of the existing shortcomings of electrochemical DNA biosensors in the field of food borne pathogen detection, we also predict and prospect future research focuses from the following five aspects: specific bioreceptors (improving specificity), nanomaterials (enhancing sensitivity), microfluidic chip technology (realizing automate operation), paper-based biosensors (reducing detection cost), and smartphones or other mobile devices (simplifying signal reading devices).


2021 ◽  
Vol 07 ◽  
Author(s):  
Wei Li

: Exploring low-cost, green and safe technologies to provide an alternative to the conventional selective catalytic reduction process is key to the control of NOx emitted from small-scale boilers and other industrial processes. To meet the demand, the chemical absorption-biological reduction integrated system has been developing recently. chemical absorption-biological reduction integrated system applies Fe(II)EDTA for NO absorption and iron-reducing and denitrifying bacteria for absorbent regeneration. Many studies have focused on the enhancements of mass transfer and biological reaction, among which the biological processes were the rate-limiting steps. This review summarizes the current researches on the biological processes in the CABR system, which focuses on the mechanism and enhancement of biochemical reactions, and provides the possible directions of future research.


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