Detection and control of environmentally transmissible viruses

Viruses, owing to their ubiquitous nature and ability to infect almost every other species, have long been a subject of interest for scientists. Some of the virus species can be very deadly to humans and animals alike and can impose a huge economic and health burden across the world. The recent CoVID-19 pandemic underscores the importance of timely detection for developing effective intervention strategies. Unfortunately, some of the virus species that cause significant health and economic impacts do not have robust and reliable detection methods due to several reasons. In some cases, despite having gold standard methods for detection of viruses, lack of effective upstream sample preparation steps could result in underestimating the viral loads. Sample preparation prior to detection is an often-overlooked aspect of foodborne virus detection. The sample preparation step is very crucial especially when food and environmental samples are involved due to the small number of infectious virus particles in a large volume of sample (eg. Fresh produce and sewage). Earlier studies have shown that representative gut bacteria strains can capture human norovirus from environmental samples. But the capture efficiency is largely dependent on the culture media conditions. The current study focuses on this aspect of sample concentration prior to detection using engineered bacterial strains. We have demonstrated that using engineered bacterial strains could effectively improve the capture efficiency of human norovirus particles from stool samples. We noticed an upwards of 65% capture efficiency with all the engineered clones we tested. This is much higher compared to that of conventional PEG or magnetic bead-based methods wherein the capture efficiencies are E. coli-based capture method can be scaled up to accommodate larger sample volumes. The engineered E. coli-based capture and concentration technique is also not susceptible to change in media conditions as the inducible expression of norovirus specific peptides expressed on the surface can be fine-tuned. This is the first time ever someone has used engineered E. coli for capture and concentration of human norovirus from environment samples. Moreover, the ease with which the engineered bacteria can be cultured and utilized for capture of norovirus makes it an ideal method for sample concentration prior to detection in resource limited settings. Control of environmentally transmissible viruses is an important aspect from a public health standpoint. To achieve this, conventional disinfection strategies employ a wide variety of chemical compounds which can often be detrimental to human health. To circumvent this issue, we propose the use of novel disinfection strategies that employ engineered water nanostructures for neutralizing both foodborne and environmental viruses. The residue-free disinfection methods proposed can be employed in a food industry setting without any problem. The EWNS cocktails used in this study showed more efficacy against a coronavirus surrogate and vegetative bacteria than MS2. Miniscule amounts of active ingredients were required to achieve inactivation of pathogens on high touch surfaces. Targeted and precise delivery of active ingredients is superior to conventional “wet” treatments. With the EWNS system, we were able to achieve complete inactivation of the SARS-CoV-2 surrogate HCoV229E after just 1 minute exposure. This demonstrates the potential of the EWNS system as an effective method for inactivating viruses on surface. The potential of EWNS for air disinfection is currently being tested. We also highlight the importance of using UV-C based disinfection methods for combating environmentally significant viruses. We tested the efficacy of 4 different commercially available UV-C light-based systems for their disinfection capacity. The two handheld devices we tested lived up to their claims of disinfecting viruses on surfaces. The airborne inactivation results show promise for occupational deployment of ceiling-based UVC 222 nm technology for a high level of SARS-CoV-2 inactivation in the air within a short time. But the potential for UV-C based disinfection techniques requires further scrutiny

Turicibacter fermentation enhances the inhibitory effects of Antrodia camphorata supplementation on tumorigenic serotonin and Wnt pathways and promotes ROS-mediated apoptosis of Caco-2 cells

Introduction: Diet-induced obesity has been shown to decrease the abundance of Turicibacter, a genus known to play a role in the serotonin signaling system, which is associated with colorectal tumorigenesis, making the presence of Turicibacter potentially influential in the protection of intestinal tumorigenesis. Recently, Antrodia camphorata (AC), a medicinal fungus native to Taiwan, has emerged as a promising candidate for complementary and alternative cancer therapy. Small molecules and polysaccharides derived from AC have been reported to possess health-promoting effects, including anti-cancer properties.Methods: Bacterial culture followed with cell culture were used in this study to determine the role of Turicibacter in colorectal tumorigenesis and to explore the anti-cancer mechanism of AC with Turicibacter fermentation.Results:Turicibacter fermentation and the addition of AC polysaccharide led to a significant increase in the production of nutrients and metabolites, including α-ketoglutaric acid and lactic acid (p < 0.05). Treatment of Turicibacter fermented AC polysaccharide was more effective in inhibiting serotonin signaling-related genes, including Tph1, Htr1d, Htr2a, Htr2b, and Htr2c (p < 0.05), and Wnt-signaling related protein and downstream gene expressions, such as phospho-GSK-3β, active β-catenin, c-Myc, Ccnd1, and Axin2 (p < 0.05). Additionally, it triggered the highest generation of reactive oxygen species (ROS), which activated PI3K/Akt and MAPK/Erk signaling and resulted in cleaved caspase-3 expression. In comparison, the treatment of AC polysaccharide without Turicibacter fermentation displayed a lesser effect.Discussion: Our findings suggest that AC polysaccharide effectively suppresses the tumorigenic serotonin and Wnt-signaling pathways, and promotes ROS-mediated apoptosis in Caco-2 cells. These processes are further enhanced by Turicibacter fermentation

Inactivating SARS-CoV-2 Surrogates on Surfaces Using Engineered Water Nanostructures Incorporated with Nature Derived Antimicrobials

The continuing cases of COVID-19 due to emerging strains of the SARS-CoV-2 virus underscore the urgent need to develop effective antiviral technologies. A crucial aspect of reducing transmission of the virus is through environmental disinfection. To this end, a nanotechnology-based antimicrobial platform utilizing engineered water nanostructures (EWNS) was utilized to challenge the human coronavirus 229E (HCoV-229E), a surrogate of SARS-CoV-2, on surfaces. The EWNS were synthesized using electrospray and ionization of aqueous solutions of antimicrobials, had a size in the nanoscale, and contained both antimicrobial agents and reactive oxygen species (ROS). Various EWNS were synthesized using single active ingredients (AI) as well as their combinations. The results of EWNS treatment indicate that EWNS produced with a cocktail of hydrogen peroxide, citric acid, lysozyme, nisin, and triethylene glycol was able to inactivate 3.8 logs of HCoV-229E, in 30 s of treatment. The delivered dose of antimicrobials to the surface was measured to be in pico to nanograms. These results indicate the efficacy of EWNS technology as a nano-carrier for delivering a minuscule dose while inactivating HCoV-229E, making this an attractive technology against SARS-CoV-2

Geldanamycin-inspired compounds induce direct trans-differentiation of human mesenchymal stem cells to neurons

Inspired from geldanamycin, the synthesis of a new series of 20-membered macrocyclic compounds is developed. The key features in our design are (i) retention of the fragment having the precise chiral functional groups of geldanamycin at C10, C11, C12 and C14, and (ii) replacement of an olefin moiety with the ester group, and the quinoid sub-structure with the triazole ring. The southern fragment needed for the macrocyclic ring formation was obtained from Evans' syn aldol as the key reaction and with the use of D-mannitol as the cheap source of a chiral starting material. For the synthesis of the northern fragment, we utilized L-ascorbic acid, which provided the desired chiral functional groups at C6 and C7. Further, the chain extension completed the synthesis of the northern fragment. In our approach, the crucial 20 membered macrocyclic ring was formed employing the click chemistry. When tested for their ability to directly trans-differentiate human mesenchymal stem cells to neurons, two novel compounds (20a and 7) from this series were identified and this was further validated by the presence of specific neuronal biomarkers (i.e. nestin, agrin and RTN4)