Asymmetry in Synergistic Interaction Between Wheat streak mosaic virus and Triticum mosaic virus in Wheat.

Wheat streak mosaic virus (WSMV) and Triticum mosaic virus (TriMV), distinct members in the family Potyviridae, are economically important wheat-infecting viruses in the Great Plains region. Previously, we reported that coinfection of wheat by WSMV and TriMV caused disease synergism with increased concentration of both viruses. The mechanisms of synergistic interaction between WSMV and TriMV and the effects of prior infection of wheat by either of these "synergistically interacting partner" (SIP) viruses on the establishment of local and systemic infection by the other SIP virus are not known. In this study, using fluorescent protein-tagged viruses, we found that prior infection of wheat by WSMV or TriMV negatively affected the onset and size of local foci elicited by subsequent SIP virus infection compared with those in buffer-inoculated wheat. These data revealed that prior infection of wheat by an SIP virus has no measurable advantage for another SIP virus on the initiation of infection and cell-to-cell movement. In TriMV-infected wheat, WSMV exhibited accelerated long-distance movement and increased accumulation of genomic RNAs compared with those in buffer-inoculated wheat, indicating that TriMV-encoded proteins complemented WSMV for efficient systemic infection. In contrast, TriMV displayed delayed systemic infection in WSMV-infected wheat, with fewer genomic RNA copies in early stages of infection compared with those in buffer-inoculated wheat. However, during late stages of infection, TriMV accumulation in WSMV-infected wheat increased rapidly with accelerated long-distance movement compared with those in buffer-inoculated wheat. Taken together, these data suggest that interactions between synergistically interacting WSMV and TriMV are asymmetrical; thus, successful establishment of synergistic interaction between unrelated viruses will depend on the order of infection of plants by SIP viruses.

Wheat streak mosaic virus alters the transcriptome of its vector, wheat curl mite (Aceria tosichella Keifer), to enhance mite development and population expansion.

Wheat streak mosaic virus (WSMV; genus Tritimovirus; family Potyviridae) is an economically important wheat virus that is transmitted by the wheat curl mite (WCM; Aceria tosichella Keifer) in a persistent manner. Virus-vector coevolution may potentially influence vector gene expression to prolong viral association and thus increase virus transmission efficiency and spread. To understand the transcriptomic responses of WCM to WSMV, RNA sequencing was performed to assemble and analyse transcriptomes of WSMV viruliferous and aviruliferous mites. Among 7291 de novo-assembled unigenes, 1020 were differentially expressed between viruliferous and aviruliferous WCMs using edgeR at a false discovery rate ≤0.05. Differentially expressed unigenes were enriched for 108 gene ontology terms, with the majority of the unigenes showing downregulation in viruliferous mites in comparison to only a few unigenes that were upregulated. Protein family and metabolic pathway enrichment analyses revealed that most downregulated unigenes encoded enzymes and proteins linked to stress response, immunity and development. Mechanistically, these predicted changes in mite physiology induced by viral association could be suggestive of pathways needed for promoting virus-vector interactions. Overall, our data suggest that transcriptional changes in viruliferous mites facilitate prolonged viral association and alter WCM development to expedite population expansion, both of which could enhance viral transmission.

Neighborhood food environment and obesity in community-dwelling older adults: individual and neighborhood effects.

OBJECTIVES: We tested hypotheses about the relationship between neighborhood-level food sources and obesity, controlling for individual-level characteristics. METHODS: Data (collected November 2006-April 2008) derived from a random-digit-dial sample of 5688 community-dwelling adults aged 50 to 74 years residing in 1644 census tracts in New Jersey. Using multilevel structural equation models, we created latent constructs representing density of fast-food establishments and storefronts (convenience stores, bars and pubs, grocery stores) and an observed indicator for supermarkets at the neighborhood level, simultaneously modeling obesity and demographic characteristics (age, gender, race, education, household income) at the individual level. RESULTS: When we controlled for individual-level age, gender, race, education, and household income, densities of fast-food establishments and storefronts were positively associated with obesity. Supermarkets were not associated with obesity. CONCLUSIONS: Because people living in neighborhoods with a higher density of fast food and storefronts are more likely to be obese, these neighborhoods may be optimal sites for interventions.

The effects of wearing a mask on an exercise regimen.

CONTEXT: Masks have long been utilized to prevent the spread of airborne pathogens and diseases in the healthcare setting. Recently, due to the COVID-19 pandemic, mask use has been expanded to all public areas to help slow the spread of this virus. One such location where masks can be seen is gyms. While exercising, the needs of the body are altered due to the increased stress being placed upon it. Normal physiology is thus adjusted to meet these new demands and to maintain optimal functioning. Therefore, it is possible that adding a mask covering the mouth and nose while exercising could further exacerbate this physiologic alteration, causing potential concerns. OBJECTIVES: The goal of this study is to identify the impact of mask use on normal perceived physiology (breathing, heart rate, temperature, exertion, stamina, and quality of workout) within the exercising population. METHODS: To obtain data focused on the research question, a self-reporting, online, anonymous Qualtrics survey was administered in local gyms and social media outlets. A total of 280 total participants were recruited between the ages of 18 and 65 who have ever exercised while wearing a mask. All results were analyzed utilizing descriptive statistics, bivariate correlations, Mann-Whitney U tests, and Kruskal-Wallis tests. A Cronbach's alpha was also calculated to check internal validity. The significance level utilized was p≤0.05. RESULTS: Completion of a Kruskal-Wallis test revealed statistical significance regarding the perception of masks in general and the participants' rating of the perceived physiological parameters (breathing: p<0.001; heart rate: p<0.001; temperature: p<0.001; exertion: p<0.001; stamina: p<0.001; and quality of workout: p<0.001), the duration of time the mask was utilized during the workout, and the participants' rating of the perceived physiological parameters (breathing: p=0.001; heart rate: p=0.020; temperature: p<0.001; exertion: p<0.001; stamina: p=0.001; quality of workout: p<0.001; and perception of mask: p<0.001), and the change in the number of days that the participants exercised per week during the pandemic as well as some of the participants' ratings of the perceived physiological parameters (breathing: p=0.042; exertion: p=0.015; stamina: p=0.027; and quality of workout: p=0.016). CONCLUSIONS: Any alterations to normal physiology perception while exercising with a mask appear to be psychological and adaptive in nature. Masks alone did not contribute to the perception of their physiologic changes.

Wheat streak mosaic virus P1 Binds to dsRNAs without Size and Sequence Specificity and a GW Motif Is Crucial for Suppression of RNA Silencing.

Wheat streak mosaic virus (WSMV; genus Tritimovirus; family Potyviridae) is an economically important virus infecting wheat in the Great Plains region of the USA. Previously, we reported that the P1 protein of WSMV acts as a viral suppressor of RNA silencing. In this study, we delineated the minimal region of WSMV P1 and examined its mechanisms in suppression of RNA silencing. We found that the 25 N-terminal amino acids are dispensable, while deletion of a single amino acid at the C-terminal region completely abolished the RNA silencing suppression activity of P1. Electrophoretic mobility shift assays with in vitro expressed P1 revealed that the P1 protein formed complexes with green fluorescent protein-derived 180-nt dsRNA and 21 and 24-nt ds-siRNAs, and WSMV coat protein-specific 600-nt dsRNA. These data suggest that the P1 protein of WSMV binds to dsRNAs in a size- and sequence-independent manner. Additionally, in vitro dicing assay with human Dicer revealed that the P1 protein efficiently protects dsRNAs from processing by Dicer into siRNAs, by forming complexes with dsRNA. Sequence comparison of P1-like proteins from select potyvirid species revealed that WSMV P1 harbors a glycine-tryptophan (GW) motif at the C-terminal region. Disruption of GW motif in WSMV P1 through W303A mutation resulted in loss of silencing suppression function and pathogenicity enhancement, and abolished WSMV viability. These data suggest that the mechanisms of suppression of RNA silencing of P1 proteins of potyvirid species appear to be broadly conserved in the family Potyviridae.

RNA silencing suppression mechanisms of Triticum mosaic virus P1: dsRNA binding property and mapping functional motifs.

Triticum mosaic virus (TriMV) is the exemplar strain of the type species of the genus Poacevirus in the family Potyviridae infecting wheat in the Great Plains region of the USA. Previously, we reported that the P1 protein of TriMV is a viral suppressor of RNA silencing. Mutational analyses of P1 showed that deletion of 55 N-terminal amino acids, and a single amino acid at the C-terminus retained its ability to suppress ssGFP-induced RNA silencing. These data suggest that the N-terminal region but not the C-terminal region of P1 is flexible for suppression of RNA silencing activity. Computational analyses revealed that TriMV P1 contains LXK/RA and zinc finger motifs at the N-terminal region and a domain containing the GW motif at the C-terminal region. Mutational analysis of TriMV P1 suggested functional roles for these motifs in suppression of RNA silencing. Electrophoretic mobility shift assays with bacterially expressed P1 protein revealed that P1 binds to 180-nt and 21- and 24-nt ds-siRNAs derived from green fluorescent protein sequence. Additionally, TriMV P1 protected the 655-nt long dsRNA derived from TriMV coat protein from dicing by the human Dicer enzyme into siRNAs. Disruption of the GW motif in TriMV P1 with a W332A mutation abolished silencing suppression, pathogenicity enhancement and viability of TriMV, suggesting a functional role for the GW motif in suppression of RNA silencing.

P7 and P8 proteins of High Plains wheat mosaic virus, a negative-strand RNA virus, employ distinct mechanisms of RNA silencing suppression.

High Plains wheat mosaic virus (genus Emaravirus), an octapartite negative-sense RNA virus, encodes two RNA silencing suppressors, P7 and P8. In this study, we found that P7 and P8 efficiently delayed the onset of dsRNA-induced transitive pathway of RNA silencing. Electrophoretic mobility shift assays (EMSA) revealed that only P7 protected long dsRNAs from dicing in vitro and bound weakly to 21- and 24-nt PTGS-like ds-siRNAs. In contrast, P8 bound strongly and relatively weakly to 21- and 24-nt ds-siRNAs, respectively, suggesting size-specific binding. In EMSA, neither protein bound to 180-nt and 21-nt ssRNAs at detectable levels. Sequence analysis revealed that P7 contains a conserved GW motif. Mutational disruption of this motif resulted in loss of suppression of RNA silencing and pathogenicity enhancement, and failure to complement the silencing suppression-deficient wheat streak mosaic virus. Collectively, these data suggest that P7 and P8 proteins utilize distinct mechanisms to overcome host RNA silencing for successful establishment of systemic infection in planta.

Octapartite negative-sense RNA genome of High Plains wheat mosaic virus encodes two suppressors of RNA silencing.

High Plains wheat mosaic virus (HPWMoV, genus Emaravirus; family Fimoviridae), transmitted by the wheat curl mite (Aceria tosichella Keifer), harbors a monocistronic octapartite single-stranded negative-sense RNA genome. In this study, putative proteins encoded by HPWMoV genomic RNAs 2-8 were screened for potential RNA silencing suppression activity by using a green fluorescent protein-based reporter agroinfiltration assay. We found that proteins encoded by RNAs 7 (P7) and 8 (P8) suppressed silencing induced by single- or double-stranded RNAs and efficiently suppressed the transitive pathway of RNA silencing. Additionally, a Wheat streak mosaic virus (WSMV, genus Tritimovirus; family Potyviridae) mutant lacking the suppressor of RNA silencing (ΔP1) but having either P7 or P8 from HPWMoV restored cell-to-cell and long-distance movement in wheat, thus indicating that P7 or P8 rescued silencing suppressor-deficient WSMV. Furthermore, HPWMoV P7 and P8 substantially enhanced the pathogenicity of Potato virus X in Nicotiana benthamiana. Collectively, these data demonstrate that the octapartite genome of HPWMoV encodes two suppressors of RNA silencing.

Molecular architecture of silk fibroin of Indian golden silkmoth, Antheraea assama.

The golden silk spun by Indian golden silkmoth Antheraea assama, is regarded for its shimmering golden luster, tenacity and value as biomaterial. This report describes the gene coding for golden silk H-fibroin (AaFhc), its expression, full-length sequence and structurally important motifs discerning the underlying genetic and biochemical factors responsible for its much sought-after properties. The coding region, with biased isocodons, encodes highly repetitious crystalline core, flanked by a pair of 5' and 3' non-repetitious ends. AaFhc mRNA expression is strictly territorial, confined to the posterior silk gland, encoding a protein of size 230 kDa, which makes homodimers making the elementary structural units of the fibrous core of the golden silk. Characteristic polyalanine repeats that make tight ß-sheet crystals alternate with non-polyalanine repeats that make less orderly antiparallel ß-sheets, ß-turns and partial α-helices. Phylogenetic analysis of the conserved N-terminal amorphous motif and the comparative analysis of the crystalline region with other saturniid H-fibroins reveal that AaFhc has longer, numerous and relatively uniform repeat motifs with lower serine content that assume tighter ß-crystals and denser packing, which are speculated to be responsible for its acclaimed properties of higher tensile strength and higher refractive index responsible for golden luster.

Current regulations and modest proposals regarding disposal of unused opioids and other controlled substances.

There are no uniform protocols in the United States for safe, environmentally acceptable disposal of controlled substances by patients. In addition, there are conflicting protocols used by various institutions for the disposal of narcotic medications. Although the US Drug Enforcement Administration oversees the prescribing, acquisition, and distribution of controlled substances and works to prevent the illegal diversion of these products, it stops short of recommending specific mechanisms for consumers to dispose of unused medications. The lack of specific regulations in this area increases the risk of illegal diversion of prescription medications and other controlled substances. The authors review and examine the dilemma posed by an ill-defined set of guidelines for disposal of controlled substances by patients and institutions not registered with the US Drug Enforcement Administration. The authors encourage public officials to update and reform ambiguous policies regarding opioid disposal by consumers and allied healthcare workers.