N-dihydrogalactochitosan reduces mortality in a lethal mouse model of SARS-CoV-2.

The rapid emergence and global dissemination of SARS-CoV-2 that causes COVID-19 continues to cause an unprecedented global health burden resulting in nearly 7 million deaths. While multiple vaccine countermeasures have been approved for emergency use, additional treatments are still needed due to sluggish vaccine rollout, vaccine hesitancy, and inefficient vaccine-mediated protection. Immunoadjuvant compounds delivered intranasally can guide non-specific innate immune responses during the critical early stages of viral replication, reducing morbidity and mortality. N-dihydrogalactochitosan (GC) is a novel mucoadhesive immunostimulatory polymer of ß-0-4-linked N-acetylglucosamine that is solubilized by the conjugation of galactose glycans with current applications as a cancer immunotherapeutic. We tested GC as a potential countermeasure for COVID-19. GC was well-tolerated and did not produce histopathologic lesions in the mouse lung. GC administered intranasally before and after SARS-CoV-2 exposure diminished morbidity and mortality in humanized ACE2 receptor expressing mice by up to 75% and reduced infectious virus levels in the upper airway. Fluorescent labeling of GC shows that it is confined to the lumen or superficial mucosa of the nasal cavity, without involvement of adjacent or deeper tissues. Our findings demonstrate a new application for soluble immunoadjuvants such as GC for preventing disease associated with SARS-CoV-2 and may be particularly attractive to persons who are needle-averse.

Exposure to diverse sarbecoviruses indicates frequent zoonotic spillover in human communities interacting with wildlife.

BACKGROUND: Sarbecoviruses are a subgenus of Coronaviridae that mostly infect bats with known potential to infect humans (SARS-CoV and SARS-CoV-2). Populations in Southeast Asia, where these viruses are most likely to emerge, have been undersurveyed to date. METHODS: We surveyed communities engaged in extractive industries and bat guano harvesting from rural areas in Myanmar. Participants were screened for exposure to sarbecoviruses, and their interactions with wildlife were evaluated to determine the factors associated with exposure to sarbecoviruses. RESULTS: Of 693 people screened between July 2017 and February 2020, 12.1% were seropositive for sarbecoviruses. Individuals were significantly more likely to have been exposed to sarbecoviruses if their main livelihood involved working in extractive industries (logging, hunting, or harvesting of forest products; odds ratio [OR] = 2.71, P = 0.019) or had been hunting/slaughtering bats (OR = 6.09, P = 0.020). Exposure to a range of bat and pangolin sarbecoviruses was identified. CONCLUSION: Exposure to diverse sarbecoviruses among high-risk human communities provides epidemiologic and immunologic evidence that zoonotic spillover is occurring. These findings inform risk mitigation efforts needed to decrease disease transmission at the bat-human interface, as well as future surveillance efforts warranted to monitor isolated populations for viruses with pandemic potential.

Severe Acute Respiratory Syndrome Coronavirus 2 Vasculopathy in a Syrian Golden Hamster Model.

Clinical evidence of vascular dysfunction and hypercoagulability as well as pulmonary vascular damage and microthrombosis are frequently reported in severe cases of human coronavirus disease 2019 (COVID-19). Syrian golden hamsters recapitulate histopathologic pulmonary vascular lesions reported in patients with COVID-19. Herein, special staining techniques and transmission electron microscopy further define vascular pathologies in a Syrian golden hamster model of human COVID-19. The results show that regions of active pulmonary inflammation in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection are characterized by ultrastructural evidence of endothelial damage with platelet marginalization and both perivascular and subendothelial macrophage infiltration. SARS-CoV-2 antigen/RNA was not detectable within affected blood vessels. Taken together, these findings suggest that the prominent microscopic vascular lesions in SARS-CoV-2-inoculated hamsters likely occur due to endothelial damage followed by platelet and macrophage infiltration.

Acute bone loss following SARS-CoV-2 infection in mice.

The novel coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and has infected more than 650 million people worldwide. Approximately 23% of these patients developed lasting "long-haul" COVID symptoms, including fatigue, joint pain, and systemic hyperinflammation. However, the direct clinical impact of SARS-CoV-2 infection on the skeletal system including bone and joint health has not been determined. Utilizing a humanized mouse model of COVID-19, this study provides the first direct evidence that SARS-CoV-2 infection leads to acute bone loss, increased osteoclast number, and thinner growth plates. This bone loss could decrease whole-bone mechanical strength and increase the risk of fragility fractures, particularly in older patients, while thinner growth plates may create growth disturbances in younger patients. Evaluating skeletal health in patients that have recovered from COVID-19 will be crucial to identify at-risk populations and develop effective countermeasures.

Sylvatic Transmission of Chikungunya Virus among Nonhuman Primates in Myanmar.

Nonhuman primates living in proximity to humans increase risks for sylvatic arbovirus transmission. We collected serum samples from nonhuman primates in Hlawga National Park near Yangon, Myanmar, and detected antibodies against chikungunya (33%) and Japanese encephalitis (4%) viruses. Buffer zones between primate and human communities might reduce cross-species arbovirus transmission.

Mucosal administration of a live attenuated recombinant COVID-19 vaccine protects nonhuman primates from SARS-CoV-2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the COVID-19 global pandemic. SARS-CoV-2 is an enveloped RNA virus that relies on its trimeric surface glycoprotein spike for entry into host cells. Here we describe the COVID-19 vaccine candidate MV-014-212, a live, attenuated, recombinant human respiratory syncytial virus expressing a chimeric SARS-CoV-2 spike as the only viral envelope protein. MV-014-212 was attenuated and immunogenic in African green monkeys (AGMs). One mucosal administration of MV-014-212 in AGMs protected against SARS-CoV-2 challenge, reducing by more than 200-fold the peak shedding of SARS-CoV-2 in the nose. MV-014-212 elicited mucosal immunoglobulin A in the nose and neutralizing antibodies in serum that exhibited cross-neutralization against virus variants of concern Alpha, Beta, and Delta. Intranasally delivered, live attenuated vaccines such as MV-014-212 entail low-cost manufacturing suitable for global deployment. MV-014-212 is currently in Phase 1 clinical trials as an intranasal COVID-19 vaccine.

Respiratory Tract Explant Infection Dynamics of Influenza A Virus in California Sea Lions, Northern Elephant Seals, and Rhesus Macaques.

To understand susceptibility of wild California sea lions and Northern elephant seals to influenza A virus (IAV), we developed an ex vivo respiratory explant model and used it to compare infection kinetics for multiple IAV subtypes. We first established the approach using explants from colonized rhesus macaques, a model for human IAV. Trachea, bronchi, and lungs from 11 California sea lions, 2 Northern elephant seals, and 10 rhesus macaques were inoculated within 24 h postmortem with 6 strains representing 4 IAV subtypes. Explants from the 3 species showed similar IAV infection kinetics, with peak viral titers 48 to 72 h post-inoculation that increased by 2 to 4 log10 PFU/explant relative to the inoculum. Immunohistochemistry localized IAV infection to apical epithelial cells. These results demonstrate that respiratory tissue explants from wild marine mammals support IAV infection. In the absence of the ability to perform experimental infections of marine mammals, this ex vivo culture of respiratory tissues mirrors the in vivo environment and serves as a tool to study IAV susceptibility, host range, and tissue tropism. IMPORTANCE Although influenza A virus can infect marine mammals, a dearth of marine mammal cell lines and ethical and logistical challenges prohibiting experimental infections of living marine mammals mean that little is known about IAV infection kinetics in these species. We circumvented these limitations by adapting a respiratory tract explant model first to establish the approach with rhesus macaques and then for use with explants from wild marine mammals euthanized for nonrespiratory medical conditions. We observed that multiple strains representing 4 IAV subtypes infected trachea, bronchi, and lungs of macaques and marine mammals with variable peak titers and kinetics. This ex vivo model can define infection dynamics for IAV in marine mammals. Further, use of explants from animals euthanized for other reasons reduces use of animals in research.

Engineering a fidelity-variant live-attenuated vaccine for chikungunya virus.

Chikungunya virus (CHIKV), which causes a febrile illness characterized by severe and prolonged polyarthralgia/polyarthritis, is responsible for a global disease burden of millions of cases each year with autochthonous transmission in over 100 countries and territories worldwide. There is currently no approved treatment or vaccine for CHIKV. One live-attenuated vaccine (LAV) developed by the United States Army progressed to Phase II human clinical trials but was withdrawn when 8% of volunteers developed joint pain associated with vaccination. Attenuation of the Army's CHIKV LAV strain 181 clone 25 (CHIKV-181/25) relies on two mutations in the envelope 2 (E2) glycoprotein responsible for cell binding and entry, making it particularly prone to reversion, a common concern for replication-competent vaccines. High error rates associated with RNA virus replication have posed a challenge for LAV development where stable incorporation of attenuating elements is necessary for establishing safety in pre-clinical models. Herein, we incorporate two replicase mutations into CHIKV-181/25 which modulate CHIKV replication fidelity combined with additional attenuating features that cannot be eliminated by point mutation. The mutations were stably incorporated in the LAV and did not increase virulence in mice. Two fidelity-variant CHIKV LAVs generated neutralizing antibodies and were protective from CHIKV disease in adult mice. Unexpectedly, our fidelity-variant candidates were more mutable than CHIKV-181/25 and exhibited restricted replication in mice and Aedes mosquitoes, a possible consequence of hypermutation. Our data demonstrate safety and efficacy but highlight a further need to evaluate fidelity-altering phenotypes before use as a LAV given the potential for virulent reversion.

Reconsidering the Role of Design Standards in Developing Effective Safety Labeling: Monolithic Recipes or Collections of Separable Features?

OBJECTIVE: This research investigated whether safety labeling design guidelines, such as the American National Standards Institute (ANSI) Z535 series, contribute to better warnings. BACKGROUND: Studies investigating the impact of safety label formatting on warning effectiveness have produced mixed findings. Additionally, research has failed to find a consistent relationship between measures of predicted and actual compliance. One commonality is that all of these studies have investigated the ANSI Z535 guidelines as a binary variable rather than as an integrative system of separable features. METHOD: We measured predicted compliance using both a within-subjects and a between-subjects design, but actual compliance using only a between-subjects design. Data were analyzed using both analysis of variance and linear/probit regressions to test the relationships between warning features recommended in the ANSI Z535 guidelines and measures of behavioral compliance. RESULTS: Predicted compliance assessed via a within-subjects design differed greatly from predicted compliance assessed via a between-subjects design. Levels of predicted and actual compliance were most similar when both measures were assessed using a between-subjects design. Consistent with previous research, location had a strong relationship with actual compliance, but surprisingly, presence of an ANSI-style orange warning header had a negative relationship with compliance. CONCLUSION: The choice of experimental design and analytical methods can dramatically influence a study's results and conclusions drawn. This research identified several aspects of experimental design that should be considered in future research on warning effectiveness. APPLICATION: Testing features recommended in the ANSI Z535 guidelines under varying conditions can contribute to the development of more effective warnings.

The Interferon-Induced Exonuclease ISG20 Exerts Antiviral Activity through Upregulation of Type I Interferon Response Proteins.

Type I interferon (IFN)-stimulated genes (ISGs) have critical roles in inhibiting virus replication and dissemination. Despite advances in understanding the molecular basis of ISG restriction, the antiviral mechanisms of many remain unclear. The 20-kDa ISG ISG20 is a nuclear 3'-5' exonuclease with preference for single-stranded RNA (ssRNA) and has been implicated in the IFN-mediated restriction of several RNA viruses. Although the exonuclease activity of ISG20 has been shown to degrade viral RNA in vitro, evidence has yet to be presented that virus inhibition in cells requires this activity. Here, we utilized a combination of an inducible, ectopic expression system and newly generated Isg20-/- mice to investigate mechanisms and consequences of ISG20-mediated restriction. Ectopically expressed ISG20 localized primarily to Cajal bodies in the nucleus and restricted replication of chikungunya and Venezuelan equine encephalitis viruses. Although restriction by ISG20 was associated with inhibition of translation of infecting genomic RNA, degradation of viral RNAs was not observed. Instead, translation inhibition of viral RNA was associated with ISG20-induced upregulation of over 100 other genes, many of which encode known antiviral effectors. ISG20 modulated the production of IFIT1, an ISG that suppresses translation of alphavirus RNAs. Consistent with this observation, the pathogenicity of IFIT1-sensitive alphaviruses was increased in Isg20-/- mice compared to that of wild-type viruses but not in cells ectopically expressing ISG20. Our findings establish an indirect role for ISG20 in the early restriction of RNA virus replication by regulating expression of other ISGs that inhibit translation and possibly other activities in the replication cycle.IMPORTANCE The host immune responses to infection lead to the production of type I interferon (IFN), and the upregulation of interferon-stimulated genes (ISGs) reduces virus replication and virus dissemination within a host. Ectopic expression of the interferon-induced 20-kDa exonuclease ISG20 suppressed replication of chikungunya virus and Venezuelan equine encephalitis virus, two mosquito-vectored RNA alphaviruses. Since the replication of alphavirus genomes occurs exclusively in the cytoplasm, the mechanism of nucleus-localized ISG20 inhibition of replication is unclear. In this study, we determined that ISG20 acts as a master regulator of over 100 genes, many of which are ISGs. Specifically, ISG20 upregulated IFIT1 genes and inhibited translation of the alphavirus genome. Furthermore, IFIT1-sensitive alphavirus replication was increased in Isg20-/- mice compared to the replication of wild-type viruses but not in cells ectopically expressing ISG20. We propose that ISG20 acts as an indirect regulator of RNA virus replication in the cytoplasm through the upregulation of many other ISGs.

Role of Place in Explaining Racial Heterogeneity in Cognitive Outcomes among Older Adults.

Racially patterned disadvantage in Southern states, especially during the formative years of primary school, may contribute to enduring disparities in adult cognitive outcomes. Drawing on a lifecourse perspective, we examine whether state of school attendance affects cognitive outcomes in older adults and partially contributes to persistent racial disparities. Using data from older African American and white participants in the national Health and Retirement Study (HRS) and the New York based Washington Heights Inwood Cognitive Aging Project (WHICAP), we estimated age-and gender-adjusted multilevel models with random effects for states predicting years of education and cognitive outcomes (e.g., memory and vocabulary). We summarized the proportion of variation in outcomes attributable to state of school attendance and compared the magnitude of racial disparities across states. Among WHICAP African Americans, state of school attendance accounted for 9% of the variance in years of schooling, 6% of memory, and 12% of language. Among HRS African Americans, state of school attendance accounted for 13% of the variance in years of schooling and also contributed to variance in cognitive function (7%), memory (2%), and vocabulary (12%). Random slope models indicated state-level African American and white disparities in every Census region, with the largest racial differences in the South. State of school attendance may contribute to racial disparities in cognitive outcomes among older Americans. Despite tremendous within-state heterogeneity, state of school attendance also accounted for some variability in cognitive outcomes. Racial disparities in older Americans may reflect historical patterns of segregation and differential access to resources such as education.

Association of proximity and density of parks and objectively measured physical activity in the United States: A systematic review.

One strategy for increasing physical activity is to create and enhance access to park space. We assessed the literature on the relationship of parks and objectively measured physical activity in population-based studies in the United States (US) and identified limitations in current built environment and physical activity measurement and reporting. Five English-language scholarly databases were queried using standardized search terms. Abstracts were screened for the following inclusion criteria: 1) published between January 1990 and June 2013; 2) US-based with a sample size greater than 100 individuals; 3) included built environment measures related to parks or trails; and 4) included objectively measured physical activity as an outcome. Following initial screening for inclusion by two independent raters, articles were abstracted into a database. Of 10,949 abstracts screened, 20 articles met the inclusion criteria. Five articles reported a significant positive association between parks and physical activity. Nine studies found no association, and six studies had mixed findings. Our review found that even among studies with objectively measured physical activity, the association between access to parks and physical activity varied between studies, possibly due to heterogeneity of exposure measurement. Self-reported (vs. independently-measured) neighborhood park environment characteristics and smaller (vs. larger) buffer sizes were more predictive of physical activity. We recommend strategies for further research, employing standardized reporting and innovative study designs to better understand the relationship of parks and physical activity.

Developing a competency framework for U.S. state food and feed testing laboratory personnel.

A competency-based training curriculum framework for U.S. state food and feed testing laboratories personnel is being developed by the International Food Protection Training Institute (IFPTI) and three partners. The framework will help laboratories catalog existing training courses/modules, identify training gaps, inform training curricula, and create career-spanning professional development learning paths, ensuring consistent performance expectations and increasing confidence in shared test results. Ultimately, the framework will aid laboratories in meeting the requirements of ISO/IEC 17025 (2005) international accreditation and the U.S. Food Safety Modernization Act (U.S. Public Law 111-353). In collaboration with the Association of Food and Drug Officials, the Association of Public Health Laboratories, and the Association of American Feed Control Officials, IFPTI is carrying out the project in two phases. In 2013, an expert panel of seven subject matter experts developed competency and curriculum frameworks for five professional levels (entry, mid-level, expert, supervisor/manager, and senior administration) across four competency domains (technical, communication, programmatic, and leadership) including approximately 80 competencies. In 2014 the expert panel will elicit feedback from peers and finalize the framework.

The impact of neighborhood park access and quality on body mass index among adults in New York City.

OBJECTIVE: To evaluate the association between adult individuals' body mass index (BMI) and characteristics of parks (size and cleanliness) in an urban environment taking into account the physical and social environments of the neighborhood. METHODS: Cross-sectional, hierarchical linear models were used to determine whether park effects were associated with BMI using self-reported height and weight data obtained from the Community Health Survey in New York City (2002-2006). RESULTS: Both the proportion of the residential zip code that was large park space and the proportion that was small park space had significant inverse associations with BMI after controlling for individual socio-demographic and zip code built environment characteristics (-0.20 BMI units across the inter-quartile range (IQR) for large parks, 95% CI -0.32, -0.08; -0.21 BMI units across the IQR for small parks, 95% CI -0.31, -0.10, respectively). Poorer scores on the park cleanliness index were associated with higher BMI, 0.18 BMI units across the IQR of the park cleanliness index (95% CI 0.05, 0.30). CONCLUSIONS: This study demonstrated that proportion of neighborhoods that was large or small park space and park cleanliness were associated with lower BMI among NYC adults after adjusting for other neighborhood features such as homicides and walkability, characteristics that could influence park usage.

Comparing nutrition environments in bodegas and fast-food restaurants.

Many small grocery stores or "bodegas" sell prepared or ready-to-eat items, filling a niche in the food environment similar to fast-food restaurants. However, little comparative information is available about the nutrition environments of bodegas and fast-food outlets. This study compared the nutrition environments of bodegas and national chain fast-food restaurants using a common audit instrument, the Nutrition Environment Measures Study in Restaurants (NEMS-R) protocol. The analytic sample included 109 bodegas and 107 fast-food restaurants located in New York City neighborhoods in the upper third and lower third of the census tract poverty rate distribution. Inter-rater reliability was evaluated in 102 food outlets, including 31 from the analytic sample and 71 from a supplementary convenience sample. The analysis compared scores on individual NEMS-R items, a total summary score, and subscores indicating healthy food availability, nutrition information, promotions of healthy or unhealthy eating, and price incentives for healthy eating, using t tests and χ(2) statistics to evaluate differences by outlet type and neighborhood poverty. Fast-food restaurants were more likely to provide nutrition information, and bodegas scored higher on healthy food availability, promotions, and pricing. Bodegas and fast-food restaurants had similar NEMS-R total scores (bodegas 13.09, fast food 14.31; P=0.22). NEMS-R total scores were higher (indicating healthier environments) in low- than high-poverty neighborhoods among both bodegas (14.79 vs 11.54; P=0.01) and fast-food restaurants (16.27 vs 11.60; P<0.01). Results imply different policy measures to improve nutrition environments in the two types of food outlets.

The development of a standards-based national curriculum framework for regulatory food safety training in the United States.

In response to the recognized need for a training system to support an integrated food safety system in the U.S., the International Food Protection Training Institute (IFPTI) in Battle Creek, Michigan, designed a career-spanning curriculum framework. IFPTI collaborated with a national curriculum team consisting of regulatory officials and university academics. The curriculum framework encompasses and organizes existing professional development for the estimated 45,000 federal, state, and local food regulators in the U.S. into efficient, standards-driven learning paths. This article describes the development process leading to an integrated national food protection training curriculum framework.

More neighborhood retail associated with lower obesity among New York City public high school students.

Policies target fast food outlets to curb adolescent obesity. We argue that researchers should examine the entire retail ecology of neighborhoods, not just fast food outlets. We examine the association between the neighborhood retail environment and obesity using Fitnessgram data collected from 94,348 New York City public high school students. In generalized hierarchical linear models, the number of fast food restaurants predicted lower odds of obesity for adolescents (OR:0.972 per establishment; CI:0.957-0.988). In a "placebo test" we found that banks--a measure of neighborhood retail ecology--also predicted lower obesity (OR:0.979 per bank; CI:0.962-0.994). Retail disinvestment might be associated with greater obesity; accordingly, public health research should study the influence of general retail disinvestment not just food-specific investment.

Neighbourhood food environments and body mass index among New York City adults.

BACKGROUND: Studies evaluating the impact of the neighbourhood food environment on obesity have summarised the density or proximity of individual food outlets. Though informative, there is a need to consider the role of the entire food environment; however, few measures of whole system attributes have been developed. New variables measuring the food environment were derived and used to study the association with body mass index (BMI). METHODS: Individual data on BMI and sociodemographic characteristics were collected from 48 482 respondents of the 2002-2006 community health survey in New York City and linked to residential zip code-level characteristics. The food environment of each zip code was described in terms of the diversity of outlets (number of types of outlets present in a zip code), the density of outlets (outlets/km(2)) and the proportion of outlets classified as BMI-unhealthy (eg, fast food, bodegas). RESULTS: Results of the cross-sectional, multilevel analyses revealed an inverse association between BMI and food outlet density (-0.32 BMI units across the IQR, 95% CI -0.45 to -0.20), a positive association between BMI and the proportion of BMI-unhealthy food outlets (0.26 BMI units per IQR, 95% CI 0.09 to 0.43) and no association with outlet diversity. The association between BMI and the proportion of BMI-unhealthy food outlets was stronger in lower (

Associations between body mass index and park proximity, size, cleanliness, and recreational facilities.

PURPOSE: To determine whether body mass index (BMI) is associated with proximity to neighborhood parks, the size of the parks, their cleanliness, and the availability of recreational facilities in the parks. DESIGN: Cross-sectional. SETTING: New York City. SUBJECTS: Adults (13,102) were recruited from 2000 to 2002 (median age 45 years, 36% male). MEASURES: Anthropometric and sociodemographic data from study subjects were linked to Department of Parks & Recreation data on park space, cleanliness, and facilities. Neighborhood-level sociodemographic and park proximity metrics were created for half-mile-radius circular buffers around each subject's residence. Proximity to park space was measured as the proportion of the subject's neighborhood buffer area that was total park space, large park space (a park > 6 acres) and small park space (a park ≤ 6 acres). Analysis. Hierarchical linear models were used to determine whether neighborhood park metrics were associated with BMI. RESULTS: Higher proximity to large park space was significantly associated with lower BMI (beta  =  -1.69, 95% confidence interval  =  -2.76, -.63). Across the population distribution of proximity to large park space, compared to subjects living in neighborhoods at the 10th percentile of the distribution, the covariate-adjusted average BMI was estimated to be .35 kg/m lower for those living in neighborhoods at the 90th percentile. The proportion of neighborhood area that was small park space was not associated with BMI, nor was park cleanliness or the availability of recreational facilities. CONCLUSION: Neighborhood proximity to large park spaces is modestly associated with lower BMI in a diverse urban population.