Comprehensive Risk Pathway of the Qualitative Likelihood of Human Exposure to Severe Acute Respiratory Syndrome Coronavirus 2 from the Food Chain.

ABSTRACT: A group of experts from all Canadian federal food safety partners was formed to monitor the potential issues relating to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) food contamination, to gather and consider all of the relevant evidence and to determine the impact for Canadian food safety. A comprehensive risk pathway was generated to consider the likelihood of a SARS-CoV-2 contamination event at any of the relevant steps of the food processing and handling chain and the potential for exposure and transmission of the virus to the consumer. The scientific evidence was reviewed and assessed for each event in the pathway, taking into consideration relevant elements that could increase or mitigate the risk of contamination. The advantage of having an event-wise contextualization of the SARS-CoV-2 transmission pathway through the food chain is that it provides a systematic and consistent approach to evaluate any new data and communicate its importance and impact. The pathway also increases the objectivity and consistency of the assessment in a rapidly evolving and high-stakes situation. Based on our review and analysis, there is currently no comprehensive epidemiological evidence of confirmed cases of SARS-CoV-2, or its known variants, causing coronavirus disease 2019 from transmission through food or food packaging. Considering the remote possibility of exposure through food, the likelihood of exposure by ingestion or contact with mucosa is considered negligible to very low, and good hygiene practices during food preparation should continue to be followed.

Cytoplasmic dynein/dynactin mediates the assembly of aggresomes.

Aggresomes are pericentrosomal cytoplasmic structures into which aggregated, ubiquitinated, misfolded proteins are sequestered. Misfolded proteins accumulate in aggresomes when the capacity of the intracellular protein degradation machinery is exceeded. Previously, we demonstrated that an intact microtubule cytoskeleton is required for the aggresome formation [Johnston et al., 1998: J. Cell Biol. 143:1883-1898]. In this study, we have investigated the involvement of microtubules (MT) and MT motors in this process. Induction of aggresomes containing misfolded DeltaF508 CFTR is accompanied by a redistribution of the retrograde motor cytoplasmic dynein that colocalizes with aggresomal markers. Coexpression of the p50 (dynamitin) subunit of the dynein/dynactin complex prevents the formation of aggresomes, even in the presence of proteasome inhibitors. Using in vitro microtubule binding assays in conjunction with immunogold electron microscopy, our data demonstrate that misfolded DeltaF508 CFTR associate with microtubules. We conclude that cytoplasmic dynein/dynactin is responsible for the directed transport of misfolded protein into aggresomes. The implications of these findings with respect to the pathogenesis of neurodegenerative disease are discussed.

A rhodopsin mutant linked to autosomal dominant retinitis pigmentosa is prone to aggregate and interacts with the ubiquitin proteasome system.

The inherited retinal degenerations are typified by retinitis pigmentosa (RP), a heterogeneous group of inherited disorders that causes the destruction of photoreceptor cells, the retinal pigmented epithelium, and choroid. This group of blinding conditions affects over 1.5 million persons worldwide. Approximately 30-40% of human autosomal dominant (AD) RP is caused by dominantly inherited missense mutations in the rhodopsin gene. Here we show that P23H, the most frequent RP mutation in American patients, renders rhodopsin extremely prone to form high molecular weight oligomeric species in the cytoplasm of transfected cells. Aggregated P23H accumulates in aggresomes, which are pericentriolar inclusion bodies that require an intact microtubule cytoskeleton to form. Using fluorescence resonance energy transfer (FRET), we observe that P23H aggregates in the cytoplasm even at extremely low expression levels. Our data show that the P23H mutation destabilizes the protein and targets it for degradation by the ubiquitin proteasome system. P23H is stabilized by proteasome inhibitors and by co-expression of a dominant negative form of ubiquitin. We show that expression of P23H, but not wild-type rhodopsin, results in a generalized impairment of the ubiquitin proteasome system, suggesting a mechanism for photoreceptor degeneration that links RP to a broad class of neurodegenerative diseases.