The key role of Calpain in COVID-19 as a therapeutic strategy.

COVID-19 is one of the viral diseases that has caused many deaths and financial losses to humans. Using the available information, this virus appears to activate the host cell-death mechanism through Calpain activation. Calpain inhibition can stop its downstream cascade reactions that cause cell death. Given the main roles of Calpain in the entry and pathogenicity of the SARS-CoV-2, its inhibition can be effective in controlling the COVID-19. This review describes how the virus activates Calpain by altering calcium flow. When Calpain was activated, the virus can enter the target cell. Subsequently, many complications of the disease, such as inflammation, cytokine storm and pulmonary fibrosis, are caused by virus-activated Calpain function. Calpain inhibitors appear to be a potential drug to control the disease and prevent death from COVID-19.

Cytotoxicity and apoptosis induction in breast cancer, skin cancer and glioblastoma cells by plant extracts.

Medicinal plants can be candidate as a common alternative for cancer treatment according to natural landscaping and native plants in each country. The aim of this study was the evaluations of cytotoxicity, apoptosis, and cell cycle arrest induction by using seven leaves extracts of Catharanthus roseus, Calystegia sepium, Berberis integerrima, Mahonia fortunei, Melia azedarach, Plantago major, Betula pendula and one bulb extract of Narcissus tazetta. Extracts were assessed on three cancer cell lines including MCF-7 breast cancer cells, A431 epidermal cell line, and U87-MG glioma cell line that were compared to HGF-1 as normal cells. According to analysis of MTT, methanolic extract of C. sepium leaves increased significantly the rate of cell death in all cancer cell lines when compared to HGF-1 as normal cells. Among different extracts, methanolic extract of C. roseus leaves and methanolic extract of C. sepium leaves indicated a crucial role in apoptosis of cancer cells according to evidences from MTT assay, cell cycle analysis, and apoptosis assay. Doxorubicin has been used as standard drug to compare with IC50 s of different extracts. In addition, the encapsulation of methanolic and ethanolic extracts in small unilamellar vesicles form (SUV) increased the cytotoxicity on cancer cell lines and normal cells. Our results indicated that different extracts can differently affect the cytotoxicity rate in variety of cancer cell lines.

Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit.

RNA and DNA extraction is a requirement for the study of gene expression and has an increasingly important role in genetic studies of all fleshy fruits. RNA and DNA extraction is difficult in kiwifruit due to the significant amount of polysaccharides and polyphenols compounds. So far, no commercial kit has been developed specifically for high-quality RNA and DNA extraction in kiwifruit and the common protocols for RNA extraction have poor yields. This study developed a new protocol for high quality RNA extraction in Actinidia deliciosa. According to the results, the average yield of RNA extraction of fruit and leaf of A. deliciosa was ∼2180.7 ng/µl (∼545.175 µg/g FW) and ∼3424.9 ng/µl (∼856.225 µg/g FW), respectively with A260/A280 between 1.95 to 2.07 and A260/A230 higher than 2 indicating high RNA purity. While the averages yield of RNA extraction using previous methods from kiwifruit and leaf was 23 µg/g FW and 527 µg/g FW, respectively. Also, the average yields of genomic DNA from kiwifruit ranged from 52 to 98 ng/µl with A260/A230 between 0.60 to 1.64 and A260/A280 between 1.40 to 1.48. To our knowledge, this is the first report of a highly efficient and rapid method of RNA and DNA extraction in kiwifruit which can be used for a broad spectrum of the all fleshy fruits.

An efficient protocol for isolation of inhibitor-free nucleic acids even from recalcitrant plants.

For fast and easy isolation of inhibitor-free genomic DNA even from the toughest plant leaf samples, including those high in polyphenols and polysaccharides, a protocol has been developed. To prevent the solubility of polysaccharides in the DNA extract, high salt concentration (1.4 M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) was used for the removal of polyphenols as polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by proteinase K and removed by centrifugation from plant extracts during the isolation process resulting in pure DNA and RNA ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA and RNA isolated from leaves and roots of recalcitrant plants which was free from contamination and color. The average yields of total RNA from roots and shoot of Betula and Grape ranged from 285 to 364 ng/µl with A260/A280 between 1.9 and 2.08. The RNA isolated with this protocol was verified to be suitable for PCR, quantitative real-time PCR, semi-quantitative reverse transcription polymerase chain reaction, cDNA synthesis and expression analysis. This protocol shown here is reproducible and can be used for a broad spectrum of plant species which have polyphenols and polysaccharide compounds.