scholarly journals Isolation and Characterization of SARS-CoV-2 strains circulating in Eastern India.

2021 ◽  
Author(s):  
Bharati Singh ◽  
KIRAN Avula ◽  
Sanchari Chatterjee ◽  
Ankita Datey ◽  
Arup Ghosh ◽  
...  

Emergence of SARS-CoV-2 as a serious pandemic has altered the global socioeconomic dynamics. The wide prevalence, high death counts and rapid emergence of new variants urge for establishment of research infrastructure to facilitate rapid development of efficient therapeutic modalities and preventive measures. In agreement with this, five SARS-CoV2 strains (ILS01, ILS02, ILS03, ILS15 and ILS24) of four different clades (19A, 19B, 20A and 20B) were isolated from patient swab samples collected during the 1st COVID-19 wave in Odisha, India. The viral isolates were adapted to in-vitro cultures and further characterized to identify strain specific variations in viral growth characteristics. All the five isolates showed substantial amount of virus induced CPE however ILS03 belonging to 20A clade displayed highest level of CPE. Time kinetics experiment revealed spike protein expression was evident after 16th hours post infection in all five isolates. ILS03 induced around 90% of cytotoxicity. Further, the susceptibility of various cell lines (human hepatoma cell line (Huh-7), CaCo2 cell line, HEK-293T cells, Vero, Vero-E6, BHK-21, THP-1 cell line and RAW 264.7 cells) were assessed. Surprisingly, it was found that the human monocyte cells THP-1 and murine macrophage cell line RAW 264.7 were permissive to all the SARS-CoV-2 isolates. The neutralization susceptibility of viral isolates to vaccine-induced antibodies was determined using sera from individuals vaccinated in the Government run vaccine drive in India. The micro-neutralization assay suggested that both Covaxin and Covishield vaccines were equally effective (100% neutralization) against all of the isolates. The whole genome sequencing of culture adapted viral isolates and viral genome from patient oropharyngeal swab sample suggested that repetitive passaging of SARS-CoV2 virus in Vero-E6 cells did not lead to emergence of many mutations during the adaptation in cell culture.  Phylogenetic analyses revealed that the five isolates clustered to respective clades. The major goal was to isolate and adapt SARS-CoV-2 viruses in in-vitro cell culture with minimal modification to facilitate research activities involved in understanding the molecular virology, host-virus interactions, application of these strains for drug discovery and animal challenge models development which eventually will contribute towards the development of effective and reliable therapeutics.

1987 ◽  
Vol 6 (3) ◽  
pp. 675-680 ◽  
Author(s):  
C.M. Chang ◽  
K.S. Jeng ◽  
C.P. Hu ◽  
S.J. Lo ◽  
T.S. Su ◽  
...  

Pharmaceutics ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 362 ◽  
Author(s):  
Amélia M. Silva ◽  
Helen L. Alvarado ◽  
Guadalupe Abrego ◽  
Carlos Martins-Gomes ◽  
Maria L. Garduño-Ramirez ◽  
...  

Oleanolic (OA) and ursolic (UA) acids are recognized triterpenoids with anti-cancer properties, showing cell-specific activity that can be enhanced when loaded into polymeric nanoparticles. The cytotoxic activity of OA and UA was assessed by Alamar Blue assay in three different cell lines, i.e., HepG2 (Human hepatoma cell line), Caco-2 (Human epithelial colorectal adenocarcinoma cell line) and Y-79 (Human retinoblastoma cell line). The natural and synthetic mixtures of these compounds were tested as free and loaded in polymeric nanoparticles in a concentration range from 2 to 32 µmol/L. The highest tested concentrations of the free triterpene mixtures produced statistically significant cell viability reduction in HepG2 and Caco-2 cells, compared to the control (untreated cells). When loaded in the developed PLGA nanoparticles, no differences were recorded for the tested concentrations in the same cell lines. However, in the Y-79 cell line, a decrease on cell viability was observed when testing the lowest concentration of both free triterpene mixtures, and after their loading into PLGA nanoparticles.


1985 ◽  
Vol 34 (6) ◽  
pp. 1017-1021 ◽  
Author(s):  
Shigehiko Uni ◽  
Masamichi Aikawa ◽  
William E. Collins ◽  
Michael R. Hollingdale ◽  
Carlos C. Campbell

2002 ◽  
Vol 76 (15) ◽  
pp. 7736-7746 ◽  
Author(s):  
Amedeo De Tomassi ◽  
Maura Pizzuti ◽  
Rita Graziani ◽  
Andrea Sbardellati ◽  
Sergio Altamura ◽  
...  

ABSTRACT Tamarins (Saguinus species) infected by GB virus B (GBV-B) have recently been proposed as an acceptable surrogate model for hepatitis C virus (HCV) infection. The availability of infectious genomic molecular clones of both viruses will permit chimeric constructs to be tested for viability in animals. Studies in cells with parental and chimeric constructs would also be very useful for both basic research and drug discovery. For this purpose, a convenient host cell type supporting replication of in vitro-transcribed GBV-B RNA should be identified. We constructed a GBV-B subgenomic selectable replicon based on the sequence of a genomic molecular clone proved to sustain infection in tamarins. The corresponding in vitro-transcribed RNA was used to transfect the Huh7 human hepatoma cell line, and intracellular replication of transfected RNA was shown to occur, even though in a small percentage of transfected cells, giving rise to antibiotic-resistant clones. Sequence analysis of GBV-B RNA from some of those clones showed no adaptive mutations with respect to the input sequence, whereas the host cells sustained higher GBV-B RNA replication than the original Huh7 cells. The enhancement of replication depending on host cell was shown to be a feature common to the majority of clones selected. The replication of GBV-B subgenomic RNA was susceptible to inhibition by known inhibitors of HCV to a level similar to that of HCV subgenomic RNA.


2003 ◽  
Vol 43 (1-3) ◽  
pp. 27-32
Author(s):  
Yutaka Miura ◽  
Miyako Ariga ◽  
Maiko Miyauchi ◽  
Katsuhiko Arai ◽  
Kazumi Yagasaki

Polymers ◽  
2021 ◽  
Vol 13 (19) ◽  
pp. 3215
Author(s):  
Gulsim Kulsharova ◽  
Akbota Kurmangaliyeva ◽  
Elvira Darbayeva ◽  
Luis Rojas-Solórzano ◽  
Galiya Toxeitova

The drug development process can greatly benefit from liver-on-a-chip platforms aiming to recapitulate the physiology, mechanisms, and functionalities of liver cells in an in vitro environment. The liver is the most important organ in drug metabolism investigation. Here, we report the development of a hybrid cyclic olefin copolymer (COC) and polydimethylsiloxane (PDMS) microfluidic (HCP) platform to culture a Huh7 hepatoma cell line in dynamic conditions towards the development of a liver-on-a-chip system. The microfluidic platform is comprised of a COC bottom layer with a microchannel and PDMS-based flat top layer sandwiched together. The HCP device was applied for culturing Huh7 cells grown on a collagen-coated microchannel. A computational fluid dynamics modeling study was conducted for the HCP device design revealing the presence of air volume fraction in the chamber and methods for optimizing experimental handling of the device. The functionality and metabolic activity of perfusion culture were assessed by the secretion rates of albumin, urea, and cell viability visualization. The HCP device hepatic culture remained functional and intact for 24 h, as assessed by resulting levels of biomarkers similar to published studies on other in vitro and 2D cell models. The present results provide a proof-of-concept demonstration of the hybrid COC–PDMS microfluidic chip for successfully culturing a Huh7 hepatoma cell line, thus paving the path towards developing a liver-on-a-chip platform.


2001 ◽  
Vol 7 (5) ◽  
pp. 685 ◽  
Author(s):  
Zhi-Qiang Song ◽  
Fei Hao ◽  
Feng Min ◽  
Qiao-Yu Ma ◽  
Guo-Dong Liu

2020 ◽  
Vol 15 (5) ◽  
pp. 1934578X2092162
Author(s):  
Kun-Min Xiao ◽  
Jun-Ming Wang ◽  
Ling-Yun Zhou ◽  
Lei Guo ◽  
Da-Cong Tan ◽  
...  

One new triterpene, 24-ene-2′-methyl carboxy acetylquercinicate, and 3 known compounds were isolated from the ethyl acetate extract of the fruiting bodies of Trametes orientalis (Yasuda) Imazeki. Their structures were elucidated by means of extensive spectroscopic analysis and compared with data reported in the literature. Furthermore, they were evaluated for their cytotoxicity against 5 human cancer cell lines in vitro. Compound 4 showed significant cytotoxicity, especially to the leukemic HL-60 cell line (IC50 = 1.77 ± 0.07 μM), while compounds 1-3 showed significant cytotoxicity to SMMC-7721 (hepatoma cell line).


Sign in / Sign up

Export Citation Format

Share Document