Beta-Cyclodextrins as affordable antivirals to treat coronavirus infection

The SARS-CoV-2 pandemic made evident that we count with few coronavirus-fighting drugs. Here we aimed to identify a cost-effective antiviral with broad spectrum activity and high safety and tolerability profiles. We began elaborating a list of 116 drugs previously used to treat other pathologies or characterized in pre-clinical studies with potential to treat coronavirus infections. We next employed molecular modelling tools to rank the 44 most promising inhibitors and tested their efficacy as antivirals against a panel of and {beta} coronavirus, e.g., the HCoV-229E and SARS-CoV-2 viruses. Four drugs, OSW-1, U18666A, hydroxypropyl-{beta}-cyclodextrin (H{beta}CD) and phytol, showed antiviral activity against both HCoV-229E (in MRC5 cells) and SARS-CoV-2 (in Vero E6 cells). The mechanism of action of these compounds was studied by transmission electron microscopy (TEM) and by testing their capacity to inhibit the entry of SARS-CoV-2 pseudoviruses in ACE2-expressing HEK-293T cells. The entry was inhibited by H{beta}CD and U18666A, yet only H{beta}CD could inhibit SARS-CoV-2 replication in the pulmonary cells Calu-3. With these results and given that cyclodextrins are widely used for drug encapsulation and can be safely administered to humans, we further tested 6 native and modified cyclodextrins, which confirmed {beta}-cyclodextrins as the most potent inhibitors of SARS-CoV-2 replication in Calu-3 cells. All accumulated data points to {beta}-cyclodextrins as promising candidates to be used in the therapeutic treatments for SARS-CoV-2 and possibly other respiratory viruses.

Multiplex Isothermal Amplification Coupled with Nanopore Sequencing for Rapid Detection and Mutation Surveillance of SARS-CoV-2

Molecular testing and surveillance of the spread and mutation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are critical public health measures to combat the pandemic. There is an urgent need for methods that can rapidly detect and sequence SARS-CoV-2 simultaneously. Here we describe a method for multiplex isothermal amplification of the SARS-CoV-2 genome in 20 minutes. Based on this, we developed NIRVANA (Nanopore sequencing of Isothermal Rapid Viral Amplification for Near real-time Analysis) to detect viral sequences and monitor mutations in multiple regions of SARS-CoV-2 genome for up to 96 patients at a time. NIRVANA uses a newly developed algorithm for on-the-fly data analysis during Nanopore sequencing. The whole workflow can be completed in as short as 3.5 hours, and all reactions can be done in a simple heating block. NIRVANA provides a rapid field-deployable solution of SARS-CoV-2 detection and surveillance of pandemic strains.

Development and Characterization of a Biodegradable PLA Food Packaging Hold Monoterpene-Cyclodextrin Complexes against Alternaria alternata.

The fungi of the genus Alternaria are among the main pathogens causing post-harvest diseases and significant economic losses. The consumption of Alternaria contaminated foods may be a major risk to human health, as many Alternaria species produce several toxic mycotoxins and secondary metabolites. To protect consumer health and extend the shelf life of food products, the development of new ways of packaging is of outmost importance. The aim of this work was to investigate the antifungal capacity of a biodegradable poly(lactic acid) (PLA) package filled with thymol or carvacrol complexed in ß-cyclodextrins (ß-CDs) by the solubility method. Once solid complexes were obtained by spray drying, varying proportions (0.0%, 1.5%, 2.5%, and 5.0 wt%) of ß-CD-thymol or ß-CD-carvacrol were mixed with PLA for packaging development by injection process. The formation of stable complexes between ß-CDs and carvacrol or thymol molecules was assessed by Fourier-transform infrared spectroscopy (FTIR). Mechanical, structural, and thermal characterization of the developed packaging was also carried out. The polymer surface showed a decrease in the number of cuts and folds as the amount of encapsulation increased, thereby reducing the stiffness of the packaging. In addition, thermogravimetric analysis (TGA) revealed a slight decrease in the temperature of degradation of PLA package as the concentration of the complexes increased, with ß-CD-carvacrol or ß-CDs-thymol complexes acting as plasticisers that lowered the intermolecular forces of the polymer chains, thereby improving the breaking point. Packages containing 2.5% and 5% ß-CD-carvacrol, or 5% ß-CD-thymol showed Alternaria alternata inhibition after 10 days of incubation revealing their potential uses in agrofood industry.

Mutations in foregut SOX2 cells induce efficient proliferation via CXCR2 pathway

Identification of the precise molecular pathways involved in oncogene-induced transformation may help us gain a better understanding of tumor initiation and promotion. Here, we demonstrate that SOX2 foregut epithelial cells are prone to oncogenic transformation upon mutagenic insults, such as Kras and p53 deletion. GFP-based lineage-tracing experiments indicate that SOX2 cells are the cells-of-origin of esophagus and stomach hyperplasia. Our observations indicate distinct roles for oncogenic KRAS mutation and P53 deletion. p53 homozygous deletion is required for the acquisition of an invasive potential, and Kras expression, but not p53 deletion, suffices for tumor formation. Global gene expression analysis reveals secreting factors upregulated in the hyperplasia induced by oncogenic KRAS and highlights a crucial role for the CXCR2 pathway in driving hyperplasia. Collectively, the array of genetic models presented here demonstrate that stratified epithelial cells are susceptible to oncogenic insults, which may lead to a better understanding of tumor initiation and aid in the design of new cancer therapeutics.