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1.
Indian J Exp Biol ; 2022 Mar; 60(3): 169-175
Article | IMSEAR | ID: sea-222467

ABSTRACT

Colorectal cancer is a one of the leading causes of death globally and its clinical management of cancer involves chemotherapy. Increase in the development of resistance to the drugs used in the cancer treatment and serious side effects associated with chemotherapeutic drugs are the major limitations in cancer therapy. Hence, there exists a huge need to develop safer natural therapeutic products for cancer therapy. In this study, ethanolic extract of Stoechospermum marginatum was evaluated for its anticancer activity. The cytotoxicity of S. marginatum extract was evaluated on HT-29 cells by MTT assay. Trypan blue cell viability was also carried out to evaluate cytotoxicity and antiproliferative effect. The apoptosis-inducing potential of the extract was analyzed by acridine orange and ethidium bromide dual staining method, mitochondrial membrane potential assay and FITC Annexin V-Propidium iodide staining method. The ethanolic extract of S. marginatum showed significant dose-dependent cytotoxicity in HT-29 cells Treatment with S. marginatum extract increased number of apoptotic cells in HT-29 cells and caused damage to mitochondrial membrane potential. The findings of the present study confirmed in vitro anticancer activity of ethanolic extract S. Marginatum

2.
Braz. j. microbiol ; 49(4): 848-855, Oct.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-974300

ABSTRACT

ABSTRACT We studied the role of Thermus thermophilus Recombinase A (RecA) in enhancing the PCR signals of DNA viruses such as Hepatitis B virus (HBV). The RecA gene of a thermophilic eubacterial strain, T. thermophilus, was cloned and hyperexpressed in Escherichia coli. The recombinant RecA protein was purified using a single heat treatment step without the use of any chromatography steps, and the purified protein (>95%) was found to be active. The purified RecA could enhance the polymerase chain reaction (PCR) signals of HBV and improve the detection limit of the HBV diagnosis by real time PCR. The yield of recombinant RecA was ∼35 mg/L, the highest yield reported for a recombinant RecA to date. RecA can be successfully employed to enhance detection sensitivity for the diagnosis of DNA viruses such as HBV, and this methodology could be particularly useful for clinical samples with HBV viral loads of less than 10 IU/mL, which is interesting and novel.


Subject(s)
Bacterial Proteins/genetics , Hepatitis B virus/isolation & purification , Polymerase Chain Reaction/methods , Thermus thermophilus/enzymology , Cloning, Molecular , Recombinases/genetics , Bacterial Proteins/isolation & purification , Bacterial Proteins/metabolism , Gene Expression , Hepatitis B virus/genetics , Polymerase Chain Reaction/instrumentation , Thermus thermophilus/genetics , Recombinases/isolation & purification , Recombinases/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism
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