Barriers and Facilitators to Online Grocery Shopping Among Supplemental Nutrition Assistance Program (SNAP) Participants: A Mixed Method Study.

This mixed methods study explored online grocery shopping perceptions by surveying individuals who do and do not receive SNAP benefits (n = 129) and by conducting interviews with SNAP recipients (n = 26) who have grocery shopped online. T-tests assessed survey findings, codebook thematic analysis was used to identify qualitative themes, and results were interpreted collectively. Survey results found no differences in perceptions of online grocery shopping between SNAP and non-SNAP recipients (p-values = 0.2-1.0) and that 97% of SNAP recipients felt comfortable using SNAP online. Five qualitative themes were identified and provided context to the survey results. The study findings can inform policy actions within SNAP.

"We're, Like, the Most Unhealthy People in the Country": Using an Equity Lens to Reduce Barriers to Healthy Food Access in Rural Appalachia.

INTRODUCTION: Obesity disproportionately affects rural communities, and Appalachia has some of the highest obesity rates in the nation. Successful policy, systems, and environmental (PSE) interventions to reduce obesity must reflect the circumstances of the population. We used a health equity lens to identify barriers and facilitators for healthy food access in Martin County, Kentucky, to design interventions responsive to social, cultural, and historical contexts. METHODS: We conducted 5 focus groups in Martin County, Kentucky, in fall 2019 to obtain perspectives on the local food system and gauge acceptability of PSE interventions. We used grounded theory to identify perceived barriers and facilitators for healthy eating. RESULTS: Thirty-four adults (27 women; median age, 46 years) participated in 5 groups. One prominent theme was declining interest in farming; many participants believed this decline was generational. One participant noted, "Most of my adult male relatives worked in the coal mines, and they worked 6 days a week. . . . My grandpa had the garden, but then my dad's generation is the one quit gardening." Another shared, "You would probably have to have someone to teach [gardening]." Instead of enhancing farmers markets, participants suggested building community capacity for home gardens to increase vegetable consumption. CONCLUSION: Our findings demonstrate the importance of obtaining community input on the development of PSE interventions to mitigate inequities in obesity. Although farmers market interventions were deemed not feasible, other solutions to enhance access to produce were identified. Developers of community-responsive PSE interventions to improve healthy eating in rural, food-insecure locations should consider using an equity-oriented prevention framework to ensure acceptable interventions.

Formation of functional, non-amyloidogenic fibres by recombinant Bacillus subtilis TasA.

Bacterial biofilms are communities of microbial cells encased within a self-produced polymeric matrix. In the Bacillus subtilis biofilm matrix, the extracellular fibres of TasA are essential. Here, a recombinant expression system allows interrogation of TasA, revealing that monomeric and fibre forms of TasA have identical secondary structure, suggesting that fibrous TasA is a linear assembly of globular units. Recombinant TasA fibres form spontaneously, and share the biological activity of TasA fibres extracted from B. subtilis, whereas a TasA variant restricted to a monomeric form is inactive and subjected to extracellular proteolysis. The biophysical properties of both native and recombinant TasA fibres indicate that they are not functional amyloid-like fibres. A gel formed by TasA fibres can recover after physical shear force, suggesting that the biofilm matrix is not static and that these properties may enable B. subtilis to remodel its local environment in response to external cues. Using recombinant fibres formed by TasA orthologues we uncover species variability in the ability of heterologous fibres to cross-complement the B. subtilis tasA deletion. These findings are indicative of specificity in the biophysical requirements of the TasA fibres across different species and/or reflect the precise molecular interactions needed for biofilm matrix assembly.

Inflammation, Immunity, and amyotrophic lateral sclerosis: I. Etiology and pathology.

Amyotrophic lateral sclerosis (ALS) is a severely debilitating disease characterized by progressive degeneration of motor neurons. Charcot first described ALS in 18691 ; however, its pathogenesis remains unknown, and effective treatments remain elusive. It is apparent that new paradigms must be investigated to understand the effectors of ALS, including inflammation, immune responses, and the body's response to stress and injury. Herein we discuss the potential role of the immune system in ALS pathogenesis and critically review evidence from patient and animal studies. Although immune system components may indeed play a role in ALS pathogenesis, studies implicating immune cells, antibodies, and cytokines in early disease pathology are limited. We propose more focused studies that examine the role of the immune system together with characterized pathogenesis to determine when, where, and if immune and inflammatory processes are critical to disease progression, and thus worthy targets of intervention. Muscle Nerve 59:10-22, 2019.

Interfacial self-assembly of a bacterial hydrophobin.

The majority of bacteria in the natural environment live within the confines of a biofilm. The Gram-positive bacterium Bacillus subtilis forms biofilms that exhibit a characteristic wrinkled morphology and a highly hydrophobic surface. A critical component in generating these properties is the protein BslA, which forms a coat across the surface of the sessile community. We recently reported the structure of BslA, and noted the presence of a large surface-exposed hydrophobic patch. Such surface patches are also observed in the class of surface-active proteins known as hydrophobins, and are thought to mediate their interfacial activity. However, although functionally related to the hydrophobins, BslA shares no sequence nor structural similarity, and here we show that the mechanism of action is also distinct. Specifically, our results suggest that the amino acids making up the large, surface-exposed hydrophobic cap in the crystal structure are shielded in aqueous solution by adopting a random coil conformation, enabling the protein to be soluble and monomeric. At an interface, these cap residues refold, inserting the hydrophobic side chains into the air or oil phase and forming a three-stranded ß-sheet. This form then self-assembles into a well-ordered 2D rectangular lattice that stabilizes the interface. By replacing a hydrophobic leucine in the center of the cap with a positively charged lysine, we changed the energetics of adsorption and disrupted the formation of the 2D lattice. This limited structural metamorphosis represents a previously unidentified environmentally responsive mechanism for interfacial stabilization by proteins.

Mutations in SIPA1L3 cause eye defects through disruption of cell polarity and cytoskeleton organization.

Correct morphogenesis and differentiation are critical in development and maintenance of the lens, which is a classic model system for epithelial development and disease. Through germline genomic analyses in patients with lens and eye abnormalities, we discovered functional mutations in the Signal Induced Proliferation Associated 1 Like 3 (SIPA1L3) gene, which encodes a previously uncharacterized member of the Signal Induced Proliferation Associated 1 (SIPA1 or SPA1) family, with a role in Rap1 signalling. Patient 1, with a de novo balanced translocation, 46,XY,t(2;19)(q37.3;q13.1), had lens and ocular anterior segment abnormalities. Breakpoint mapping revealed transection of SIPA1L3 at 19q13.1 and reduced SIPA1L3 expression in patient lymphoblasts. SIPA1L3 downregulation in 3D cell culture revealed morphogenetic and cell polarity abnormalities. Decreased expression of Sipa1l3 in zebrafish and mouse caused severe lens and eye abnormalities. Sipa1l3(-/-) mice showed disrupted epithelial cell organization and polarity and, notably, abnormal epithelial to mesenchymal transition in the lens. Patient 2 with cataracts was heterozygous for a missense variant in SIPA1L3, c.442G>T, p.Asp148Tyr. Examination of the p.Asp148Tyr mutation in an epithelial cell line showed abnormal clustering of actin stress fibres and decreased formation of adherens junctions. Our findings show that abnormalities of SIPA1L3 in human, zebrafish and mouse contribute to lens and eye defects, and we identify a critical role for SIPA1L3 in epithelial cell morphogenesis, polarity, adhesion and cytoskeletal organization.

Molecular findings from 537 individuals with inherited retinal disease.

BACKGROUND: Inherited retinal diseases (IRDs) are a clinically and genetically heterogeneous set of disorders, for which diagnostic second-generation sequencing (next-generation sequencing, NGS) services have been developed worldwide. METHODS: We present the molecular findings of 537 individuals referred to a 105-gene diagnostic NGS test for IRDs. We assess the diagnostic yield, the spectrum of clinical referrals, the variant analysis burden and the genetic heterogeneity of IRD. We retrospectively analyse disease-causing variants, including an assessment of variant frequency in Exome Aggregation Consortium (ExAC). RESULTS: Individuals were referred from 10 clinically distinct classifications of IRD. Of the 4542 variants clinically analysed, we have reported 402 mutations as a cause or a potential cause of disease in 62 of the 105 genes surveyed. These variants account or likely account for the clinical diagnosis of IRD in 51% of the 537 referred individuals. 144 potentially disease-causing mutations were identified as novel at the time of clinical analysis, and we further demonstrate the segregation of known disease-causing variants among individuals with IRD. We show that clinically analysed variants indicated as rare in dbSNP and the Exome Variant Server remain rare in ExAC, and that genes discovered as a cause of IRD in the post-NGS era are rare causes of IRD in a population of clinically surveyed individuals. CONCLUSIONS: Our findings illustrate the continued powerful utility of custom-gene panel diagnostic NGS tests for IRD in the clinic, but suggest clear future avenues for increasing diagnostic yields.

The use of autozygosity mapping and next-generation sequencing in understanding anterior segment defects caused by an abnormal development of the lens.

The formation of the anterior segment of the eye is an intricate process that is dependent to a large degree on the normal development of the lens. Despite intensive study of the role of well-described eye genes, many causes of lenticular and anterior segment anomalies remain elusive. The majority of genes implicated thus far act in an autosomal dominant manner. Autosomal recessive causes are less well described; their diagnosis has been hindered by technological limitations, extreme genetic heterogeneity, a lack of understanding of eye biology and the role of many genes within the genome. The opportunity for the discovery of extremely rare autosomal recessive causes of ocular abnormalities from the study of consanguineous families is large, particularly through the powerful combination of next-generation sequencing with autozygosity mapping. Having begun to overcome the genetic heterogeneity bottleneck, it is increasingly recognised that the interpretation of genetic variants and the association of novel genes with a particular phenotype remain challenging. Nonetheless, increasing understanding of the genetic and mutational basis of lens and anterior segment abnormalities will be of enormous value to our comprehension of eye disease(s). Further, it will improve our ability to accurately interpret putative disease-causing variants with the aim of providing more personalised patient care and avoiding lifelong visual loss in children.

Personalized diagnosis and management of congenital cataract by next-generation sequencing.

PURPOSE: To assess the utility of integrating genomic data from next-generation sequencing and phenotypic data to enhance the diagnosis of bilateral congenital cataract (CC). DESIGN: Evaluation of diagnostic technology. PARTICIPANTS: Thirty-six individuals diagnosed with nonsyndromic or syndromic bilateral congenital cataract were selected for investigation through a single ophthalmic genetics clinic. METHODS: Participants underwent a detailed ophthalmic examination, accompanied by dysmorphology assessment where appropriate. Lenticular, ocular, and systemic phenotypes were recorded. Mutations were detected using a custom-designed target enrichment that permitted parallel analysis of 115 genes associated with CC by high-throughput, next-generation DNA sequencing (NGS). Thirty-six patients and a known positive control were tested. Suspected pathogenic variants were confirmed by bidirectional Sanger sequencing in relevant probands and other affected family members. MAIN OUTCOME MEASURES: Molecular genetic results and details of clinical phenotypes were identified. RESULTS: Next-generation DNA sequencing technologies are able to determine the precise genetic cause of CC in 75% of individuals, and 85% patients with nonsyndromic CC were found to have likely pathogenic mutations, all of which occurred in highly conserved domains known to be vital for normal protein function. The pick-up rate in patients with syndromic CC also was high, with 63% having potential disease-causing mutations. CONCLUSIONS: This analysis demonstrates the clinical utility of this test, providing examples where it altered clinical management, directed care pathways, and enabled more accurate genetic counseling. This comprehensive screen will extend access to genetic testing and lead to improved diagnostic and management outcomes through a stratified medicine approach. Establishing more robust genotype-phenotype correlations will advance knowledge of cataract-forming mechanisms.

Genetic testing for inherited ocular disease: delivering on the promise at last?

Genetic testing is of increasing clinical utility for diagnosing inherited eye disease. Clarifying a clinical diagnosis is important for accurate estimation of prognosis, facilitating genetic counselling and management of families, and in the future will direct gene-specific therapeutic strategies. Often, precise diagnosis of genetic ophthalmic conditions is complicated by genetic heterogeneity, a difficulty that the so-called 'next-generation sequencing' technologies promise to overcome. Despite considerable counselling and ethical complexities, next-generation sequencing offers to revolutionize clinical practice. This will necessitate considerable adjustment to standard practice but has the power to deliver a personalized approach to genomic medicine for many more patients and enhance the potential for preventing vision loss.

Enduring Effects of the COVID-19 Pandemic on Food Access, Nutrition, and Well-Being in Rural Appalachia.

The COVID-19 pandemic produced acute effects on health inequities, yet more enduring impacts in vulnerable populations in rural Appalachia are understudied. This qualitative study included three focus groups with thirty-nine adults (74% female, mean age 52.7 years) to obtain perspectives on the impact of the COVID-19 pandemic on well-being in Martin County, Kentucky, in fall 2022. Grounded Theory was employed using an iterative inductive-deductive approach to capture the lasting effects of the COVID-19 pandemic on health practices and status. Three prominent themes emerged: (1) increased social isolation; (2) household cost of living strains caused by inflation; and (3) higher food prices and diminished food availability causing shifts in food purchasing and consumption. Participants noted that the rising cost of living resulted in residents having to "choose between medication, food and utilities". Increased food prices resulted in residents "stretching" their food, modifying how they grocery shopped, and limiting meat consumption. Persistent food shortages were exacerbated by there being few grocery stores in the county. Lastly, increased social isolation was profoundly articulated as widely impacting mental health, especially among youth. Our findings underscore the ongoing deleterious effects of inflation and food supply chain disruptions in this rural, geographically isolated community, which resulted in difficult spending choices for residents.

Household Food Sourcing Patterns and Their Associations With Food Insecurity in Appalachian Ohio.

BACKGROUND: Households experiencing food insecurity may use dynamic strategies to meet food needs. Yet, the relationship between household food sourcing behaviors and food security, particularly in rural settings, is understudied. OBJECTIVE: To identify food sourcing patterns and their associations with food insecurity among households in rural Appalachian Ohio during the COVID-19 pandemic. DESIGN: Survey data were collected from a cohort of households in Athens County, OH, in July 2020, October 2020, January 2021, and April 2021. PARTICIPANTS/SETTING: The sample included 663 households with household food sourcing and food security information for ≥1 survey wave. MAIN OUTCOME MEASURES: Household food sourcing patterns. Households reported the frequency with which they obtained food from various retailers and charitable sources, classified as supercenters, supermarkets, convenience stores, farmers markets, or charitable sources. STATISTICAL ANALYSES: Principal component analysis was used to identify food sourcing patterns. Linear mixed models were used to assess changes in food sourcing behaviors over the study period and to determine whether food sourcing behaviors differed according to food security status. RESULTS: Two patterns were identified: Convenience Stores and Charitable Food and Supermarkets and Farmers Markets, not Supercenters. Relative to July 2020, alignment of households' food sourcing behaviors with the Supermarkets and Farmers Markets, not Supercenters pattern was higher in October 2020 (ß .07, 95% CI .02 to .12) and alignment with the Convenience Stores and Charitable Food pattern was lower in April 2021 (ß -.06, 95% CI -.11 to -.02). Compared with food-secure households, food sourcing behaviors of food-insecure households were more closely aligned with the Convenience Stores and Charitable Food pattern (ß .07, 95% CI .00 to .13); no statistically significant difference in scores was observed for the Supermarkets and Farmers Markets, not Supercenters pattern (ß -.07, 95% CI -.15 to .02). CONCLUSIONS: These findings support efforts to increase access to healthy, affordable options at venues where food-insecure households may be likely to obtain food, such as convenience stores and charitable sources.

Impact of community-driven interventions on dietary and physical activity outcomes among a cohort of adults in a rural Appalachian county in Eastern Kentucky, 2019-2022.

Several environmental level factors exacerbate poor health outcomes in rural populations in the United States, such as lack of access to healthy food and locations to be physically active, which support healthy choices at the individual level. Thus, utilizing innovative place-based approaches in rural locations is essential to improve health outcomes. Leveraging community assets, like Cooperative Extension, is a novel strategy for implementing community-driven interventions. This prospective cohort study (n = 152), recruited in 2019 and surveyed again in 2020 and 2021, examined individual level changes in diet and physical activity in one rural Appalachian county. During this time, multiple community-driven interventions were implemented alongside Cooperative Extension and several community partners. Across the three-year study, the cohort indicated increases in other vegetables and water and reductions in fruits and legumes. There were also reductions in less healthy items such as French fries and sugar-sweetened beverages. The cohort also reported being less likely to engage in physical activity. Our findings suggest that key community-driven programs may have indirect effects on dietary and physical activity choices over time. Outcomes from this study are relevant for public health practitioners and community organizations working within rural Appalachian communities to address health-related behaviors.

The Impact of a Grocery Store Closure in One Rural Highly Obese Appalachian Community on Shopping Behavior and Dietary Intake.

Research has examined how the entry of grocery stores into neighborhoods influences dietary outcomes, yet limited evidence suggests a direct correlation between opening a store and changes in dietary intake. A factor that might influence individuals' behavior more directly is the closing of a grocery store where residents shop. This study aims to examine how a grocery closure in a rural Appalachian high poverty county is associated with dietary intake. A cohort of n = 152 individuals were recruited to participate in a longitudinal study examining purchasing habits and dietary intake. At time point two, one year later, n = 74 individuals completed the survey via phone. Results indicate those that switched from shopping at a local grocery store to a supercenter significantly increased their dietary intake of fruit (0.2 ± 0.8), fruits and vegetables (1.4 ± 2.7), alcohol (grams) (17.3 ± 54.1), and tomato sauce (0.1 ± 0.3). A local grocery store closure was associated with a change in shopping behavior and dietary intake. Community-level interventions targeting dietary behaviors must account for neighborhood food environment influences, including grocery store availability. Policy aimed at improving food access in rural communities need to consider approaches to improving a variety of food venues with affordable healthy food, while addressing the evolving grocery shopping behaviors of consumers.

Barriers and Facilitators of Online Grocery Services: Perceptions from Rural and Urban Grocery Store Managers.

Online grocery shopping has expanded rapidly in the U.S., yet little is known about the retailer's perceptions of online grocery services, which can aid in the expansion of services. Furthermore, many barriers to online grocery utilization persist across geographic areas, especially among Supplemental Nutrition Assistance Program (SNAP)-authorized retailers. This study captured perceived barriers and facilitators of online grocery shopping for managers of SNAP-authorized retailers. Qualitative semi-structured interviews were conducted with managers (n = 23) of grocery stores/supermarkets in urban and rural areas across four different states: TN, KY, NC, and NY. Grocery store managers offering online ordering (n = 15) and managers from brick-and-mortar stores without online services (n = 8) participated in the interviews. Three primary themes emerged among managers offering online ordering: (1) order fulfillment challenges, (2) perceived customer barriers, and (3) perceived customer benefits. Among managers at brick-and-mortar locations without online services, four major themes emerged: (1) thoughts on implementing online shopping, (2) COVID-19 pandemic impacts, (3) competition with other stores, and (4) benefits of maintaining brick-and-mortar shopping. This study provides a deeper understanding of retailers' experience and perceptions of online grocery services among stores authorized to accept SNAP benefits. This perspective is necessary to inform policies and enhance the evolving virtual food marketplace for SNAP customers.

Online Pilot Grocery Intervention among Rural and Urban Residents Aimed to Improve Purchasing Habits.

Online grocery shopping has the potential to improve access to food, particularly among low-income households located in urban food deserts and rural communities. The primary aim of this pilot intervention was to test whether a three-armed online grocery trial improved fruit and vegetable (F&V) purchases. Rural and urban adults across seven counties in Kentucky, Maryland, and North Carolina were recruited to participate in an 8-week intervention in fall 2021. A total of 184 adults were enrolled into the following groups: (1) brick-and-mortar "BM" (control participants only received reminders to submit weekly grocery shopping receipts); (2) online-only with no support "O" (participants received weekly reminders to grocery shop online and to submit itemized receipts); and (3) online shopping with intervention nudges "O+I" (participants received nudges three times per week to grocery shop online, meal ideas, recipes, Facebook group support, and weekly reminders to shop online and to submit itemized receipts). On average, reported food spending on F/V by the O+I participants was USD 6.84 more compared to the BM arm. Online shopping with behavioral nudges and nutrition information shows great promise for helping customers in diverse locations to navigate the increasing presence of online grocery shopping platforms and to improve F&V purchases.

Mechanical, sensory, and consumer evaluation of ketogenic, gluten-free breads.

Ketogenic, gluten-free breads comprised of almond flour, oat bran fiber, or combinations of both were compared. The textural properties, sensory attributes, and consumer acceptance were analyzed on each bread containing 100% almond flour (AF), 66.7% almond flour with 33.3% oat bran fiber (AOB), 66.7% oat bran fiber with 33.3% almond flour (OBA), and 100% oat bran fiber (OB). AF and AOB breads had a more open crumb structure composed of cells between 1-4 mm2. OBA and OB had a significantly dense crumb pattern made up of more cells less than one millimeter squared. Quantitative-descriptive analysis (QDA) and consumer acceptance testing was conducted 24 hr after baking and mechanical endpoints were evaluated 24, 72, and 120 hr after baking. AF and AOB breads were preferred over OBA and OB breads in QDA evaluation and consumer acceptance scores. Greater percentages of oat bran fiber resulted in a bread that was less moist, firmer in texture, and chewier with trained panelists. In both sensory evaluations, higher amounts of almond flour resulted in higher values in eggy flavor while increased amounts of oat bran fiber correlated with higher values in earthy flavor. Mechanical testing identified higher percentages of almond flour resulted in bread that was less firm and less chewy. Over time, all variations with almond flour became softer and less chewy, while the OB bread increased in firmness. Sensory cohesiveness did not correlate with the mechanical equivalent, identifying a need to re-evaluate the parameters used to calculate this objective endpoint.

Community-Based Efforts Aim to Improve the Food Environment within a Highly Obese Rural Appalachian County.

Rural communities in Appalachia are displaying increased obesity prevalence, yet traditional interventions have not provided a broad enough impact to improve dietary consumption patterns. Therefore, expanding efforts that address the food environment and incorporate behavioral nudges through community-developed marketing strategies may be a viable mechanism to improve food and beverage choices within this unique population. This study installed shelf-wobblers across n = 5 gas stations in one rural Appalachian county in Kentucky. Smart Snacks were identified from store inventory lists utilizing the CDC Food Service Guideline for Federal Facilities calculator and were categorized into high-protein snacks, low-fat carbohydrate snacks, meal replacement snacks, and no-calorie beverages. NEMS-CS audits were conducted, and monthly sales data was collected at baseline and for six months thereafter for each store location. A difference-in-difference model was used, adjusting for total sales or total mean sales for each Smart Snack model to assess the percentage change within and between stores. Overall, percent change in mean sales and total sales across all stores resulted in a percentage increase of sales of Smart Snack items following wobbler installment. This study provides unique insight into how a community-driven approach to marketing can influence the sale of healthier food and beverage items.

Sugar-sweetened beverage consumption among adults in rural Appalachia.

Sugar-sweetened beverage (SSB) consumption is decreasing nationally, yet intakes remain high in certain sub-populations as new varieties of SSBs are introduced. This study aims to expand on SSB intake patterns among adults living in Appalachia to develop policy, systems, and environmental (PSE) interventions to reduce consumption. Baseline cohort surveys were conducted to examine beverage consumption patterns of adults in one rural Appalachian county in Kentucky using a validated BEVQ-15 instrument. Ages were collapsed into three generational groups - Millennials (22-38 years), Generation X (39-54 years), and Boomers/Silents (≥55 years). Over half (n = 81; 54%) of the sample (n = 150) were Boomers/Silents. Age was a significant predictor of SSB consumption, with Millennials drinking more daily calories of SSB compared to older adults (329.2 kcal v 157.0 kcal v 134.6 kcal, p = 0.05); a significant amount of those calories coming from non-soda SSBs. Millennials were twice as likely to drink sweetened fruit juice drinks (p = 0.0002) and energy drinks (p = 0.01) daily and consumed six times more daily calories from sweetened fruit juice drinks than the other groups (73.5 kcal v 11.1 kcal v 8.0 kcal, p < 0.01). To our knowledge, this is the first study to show beverage choices and consumption patterns in Appalachian adults vary by age and non-soda SSBs are significant sources of added sugar. These findings inform PSE interventions for reducing SSB consumption, such as tailored marketing approaches and technology-based strategies, within a unique setting, and offer insight for nutrition educators and public health professionals working within rural, remote communities.