Hepato-protective, antioxidant activities and acute toxicity of a stem bark extract of Erythrina senegalensis DC.

This study aimed at evaluating the in vitro antioxidant and hepatoprotective activities of different stem bark extracts of Erythrina senegalensis prepared with ethanol, and the in vivo hepatoprotective activity and acute toxicity of the best extract. The 2, 4-diphenyl-1-picryl-hydrazil (DPPH) and microsomal lipid peroxidation (MLP) models, and the rat liver slices system were respectively used for the in vitro study. The Methylene chloride/methanol (1:1 v/v) (Emc) and 40% ethanolic (E40) extracts were more efficient in inhibiting MLP and in scavenging DPPH radical. However, E40 was most effective with regards to lactate dehydrogenase (LDH) leakage inhibition from rat liver slices intoxicated with carbon tetrachloride (CCl4). The in vivo hepatoprotective activity was evaluated against CCl4-induced hepatotoxicity in rats. The E40 extract (100 mg/Kg) significantly reduced the increase in ALT, AST and lipid peroxidation in liver homogenate, showing that the extract is as protective as silymarin at the same dose. Acute toxicity was evaluated in mice and E40 did not produce any behavioural changes or mortality even at an oral dose of 16 g/kg. The extract was found to contain antioxidant classes of compounds (flavonoids and polyphenols). In conclusion, the E40 extract of E. senegalensis could be an important source of hepatoprotective compounds.Key words: Erythrina senegalensis, stem bark extract, antioxidant, hepatoprotective, carbon tetrachloride

Recommendations for accurate genotyping of SARS-CoV-2 using amplicon-based sequencing of clinical samples.

Genotyping of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been instrumental in monitoring viral evolution and transmission during the pandemic. The quality of the sequence data obtained from these genotyping efforts depends on several factors, including the quantity/integrity of the input material, the technology, and laboratory-specific implementation. The current lack of guidelines for SARS-CoV-2 genotyping leads to inclusion of error-containing genome sequences in genomic epidemiology studies. We aimed to establish clear and broadly applicable recommendations for reliable virus genotyping. We established and used a sequencing data analysis workflow that reliably identifies and removes technical artefacts; such artefacts can result in miscalls when using alternative pipelines to process clinical samples and synthetic viral genomes with an amplicon-based genotyping approach. We evaluated the impact of experimental factors, including viral load and sequencing depth, on correct sequence determination. We found that at least 1000 viral genomes are necessary to confidently detect variants in the SARS-CoV-2 genome at frequencies of ≥10%. The broad applicability of our recommendations was validated in over 200 clinical samples from six independent laboratories. The genotypes we determined for clinical isolates with sufficient quality cluster by sampling location and period. Our analysis also supports the rise in frequencies of 20A.EU1 and 20A.EU2, two recently reported European strains whose dissemination was facilitated by travel during the summer of 2020. We present much-needed recommendations for the reliable determination of SARS-CoV-2 genome sequences and demonstrate their broad applicability in a large cohort of clinical samples