Design of an Ara h 2 hypoallergen from conformational epitopes.

INTRODUCTION: Adverse reactions are relatively common during peanut oral immunotherapy. To reduce the risk to the patient, some researchers have proposed modifying the allergen to reduce IgE reactivity, creating a putative hypoallergen. Analysis of recently cloned human IgG from patients treated with peanut immunotherapy suggested that there are three common conformational epitopes for the major peanut allergen Ara h 2. We sought to test if structural information on these epitopes could indicate mutagenesis targets for designing a hypoallergen and evaluated the reduction in IgE binding via immunochemistry and a mouse model of passive cutaneous anaphylaxis (PCA). METHODS: X-ray crystallography characterized the conformational epitopes in detail, followed by mutational analysis of key residues to modify monoclonal antibody (mAb) and serum IgE binding, assessed by ELISA and biolayer interferometry. A designed Ara h 2 hypoallergen was tested for reduced vascularization in mouse PCA experiments using pooled peanut allergic patient serum. RESULTS: A ternary crystal structure of Ara h 2 in complex with patient antibodies 13T1 and 13T5 was determined. Site-specific mutants were designed that reduced 13T1, 13T5, and 22S1 mAbs binding by orders of magnitude. By combining designed mutations from the three major conformational bins, a hexamutant (Ara h 2 E46R, E89R, E97R, E114R, Q146A, R147E) was created that reduced IgE binding in serum from allergic patients. Further, in the PCA model where mice were primed with peanut allergic patient serum, reactivity upon allergen challenge was significantly decreased using the hexamutant. CONCLUSION: These studies demonstrate that prior knowledge of common conformational epitopes can be used to engineer reduced IgE reactivity, an important first step in hypoallergen design.

The Emerging Leaders in Public Health Initiative: Individual Leadership Transformation.

The Emerging Leaders in Public Health (ELPH) Initiative, sponsored by The Kresge Foundation, provided leadership development to local governmental public health leaders. An adaptive leadership framework was used to shape the curriculum. The coleads attended multiday convenings and webinars over a 16- to 18-month period. Key components of the initiative included using applied learning to solidify their leadership skills as they worked to develop a new role for their agencies provision of a resource grant from The Kresge Foundation to support agency transformation and technical assistance and consultation provided by a National Program Office. An external evaluator conducted a multicomponent evaluation of individual leadership skill change. Graduates assessed change in their own leadership and change in their colead. Colleagues of ELPH participants were surveyed about their observed changes in the leadership actions of the program graduates. One hundred four leaders from 30 states participated in the initiative in 3 successive cohorts. Leaders demonstrated improvement as shown by self-report and external observation. Improved ability to communicate in a way that inspires others was one of the most significant changes in leadership behaviors. Additional leadership actions were strengthened including the ability to create and maintain high-performing teams, the ability to ask transformational questions, and the ability to actively listen to understand. The pandemic has shown the importance of developing the field, starting with leadership. Leadership development and agency transformation are synergistic; they depend on each other and support each other.

The Emerging Leaders in Public Health Initiative: Leadership Development to Advance Public Health Practice.

CONTEXT: Effective leadership is essential for governmental public health agencies to promote and protect the health of their communities. OBJECTIVE: The Emerging Leaders in Public Health Initiative, a program of The Kresge Foundation, was implemented to strengthen leadership in governmental public health. We seek to examine lessons from the initiative to contribute to the field a better understanding of leadership development practices. DESIGN: An external evaluator conducted a retrospective analysis of participant responses postinitiative to assess the overall impact of the initiative and explore which components were most valuable. SETTING: United States. PARTICIPANTS: Directors and other staff from governmental public health agencies were recruited in 2-person teams to participate in 3 successive cohorts. INTERVENTION: Drawn from adaptive leadership principles, a framework was developed to guide the selection and implementation of educational and experiential activities. Participants were challenged to develop a new role for their public health agency, which provided a learning laboratory to practice individual and team leadership skills. The Kresge Foundation provided participants with a resource grant and a National Program Office offered convenings, webinars, coaching, and technical assistance over the 18 months of the developmental experience. MAIN OUTCOME MEASURES: Satisfaction, perceived value of components, and future intentions were assessed from participants in cohorts II and III (n = 70). The overall response rate was 93%. RESULTS: One hundred four diverse leaders from 52 agencies, representing 30 states participated in the initiative. Participants were extremely satisfied with the program (94%) and indicated a strong likelihood of recommending it to a colleague (96%). Unrestricted grant funding, peer learning, and in-person learning sessions were the program components rated most highly valuable. CONCLUSIONS: This initiative offers insights into principles and processes to be considered for future public health leadership development.

The role of stress responses on engagement in dietary and physical activity behaviors among Latino adults living with prediabetes.

OBJECTIVES: Latinos are at a high risk of developing type 2 diabetes (T2D). Prediabetes is a major risk factor for T2D; however, progression to T2D can be slowed with engagement in healthy behaviors. Stress can hinder engagement with health behaviors. Qualitative methods were used to understand how Latinos with prediabetes attempted to modify their diet and physical activity behaviors to slow T2D progression and how stress affected their engagement in these behaviors. DESIGN: Semi-structured interviews were conducted with 20 Latinos with prediabetes in North Carolina. Participants were asked questions about types of stress they experienced and how stress affected their health. We generated codes on stress and stress responses and used content analysis to organize codes between and within participants. RESULTS: Behaviors changed after prediabetes diagnosis. Few participants reported changing their physical activity, however, all participants attempted to change their eating patterns by changing food types consumed and reducing portion sizes. The stress participants experienced impacted their ability to self-regulate their diet. They reported overeating or appetite suppression during stressful periods. Stress also affected cognitive responses by compromising healthy decision-making and instigating negative emotional reactions. Overall, stress complicated participants' ability to properly engage in recommended behaviors by negatively impacting participants' behavioral self-regulation and cognitive processes. CONCLUSIONS: Stress affects behavioral and cognitive progresses that adversely alters primarily dietary behaviors. Tailored plans acknowledging the impact of stress and providing coping and supportive help for dealing with stress may enhance engagement in healthy behaviors for Latinos with prediabetes.

An assessment of the Libyan baccalaureate nursing education during political turmoil.

Since the Libyan Revolution in 2011, the country's nursing workforce has been struggling. Libyan nursing schools have focused on rebuilding the country's supply of nurses after many emigrated. Wanting to infuse the workforce with more baccalaureate-prepared nurses, Libyan nursing faculty invited nursing and public health representatives from a US-based academic medical institution and a non-governmental medical organization to collaborate with local stakeholders in a country-wide assessment. The purpose of this article is to outline the national programs' strengths and weaknesses and make recommendations for developing a strategy to elevate nursing education to meet international standards. This can serve as a launching point to strengthen Libya's health services provision capacity, particularly during this time of transition when opportunities may become available to move in new directions. The approach and findings may have wider application to other countries who are similarly experiencing civil and political turmoil.

Bridging the Urban-Rural Divide in Chronic Disease Through Community Engagement in Health Program Delivery.

The urban-rural divide in chronic disease contributes to persistent geographic disparities in life expectancy in North Carolina. Policies and programs in resource-constrained rural areas should be designed to increase health equity and reduce this continuing divide by including greater community engagement and decision-making for meaningful, sustainable change.

Action Learning Cohort Series: An Innovative Community-Engaged Approach for Translating Research Into Practice.

Cultivating strong partnerships among community and academic stakeholders expedites the translation of research findings into practice and communities by enhancing opportunities for research dissemination and implementation. However, the lack of systematic methods for community stakeholder engagement may decelerate the translational research process. The North Carolina Translational Research and Clinical Sciences Institute implemented an innovative approach to community engagement called the Action Learning Cohort (ALC) Series. The ALC Series, a workgroup extension of a professional conference, used action learning and systems thinking strategies to conceptualize and develop a product aimed at preventing, treating, and controlling hypertension in eastern North Carolina. We evaluated the acceptability and practicality of the ALC Series using survey, focus group, and interview pilot data. Action learning and systems thinking strategies led ALC stakeholders to develop and disseminate The Empathy Building Resource Guide: A Toolkit for Enhancing Patient-Provider Relationships in the Treatment, Management, and Prevention of Hypertension. Stakeholders rated the Series as satisfactory and acknowledged gains in knowledge and desire for engagement with fellow ALC stakeholders beyond the Series. The ALC Series approach is a potentially practical and acceptable model for systematically engaging community stakeholders in translating knowledge into a product that addresses health topics like hypertension.

Identification of the amino-terminal fragment of Ara h 1 as a major target of the IgE-binding activity in the basic peanut protein fraction.

BACKGROUND: Small, basic peanut proteins are often poorly extracted in pH-neutral buffers that are optimal for the extraction of peanut storage proteins such as Ara h 1. As a result, such proteins are easily missed as potential allergens. OBJECTIVE: To analyse the allergenic composition of the basic peanut protein (BPP) fraction. METHODS: A peanut extract prepared at pH 4 was fractionated by physicochemical procedures. Chemical analysis was performed by SDS-PAGE and mass spectrometry. Because immunoblotting was found to be inefficient for most of these small basic proteins, IgE-binding activity was measured by coupling the fractions to CNBr-activated Sepharose, followed by incubation with sera from 55 Dutch peanut-allergic children and 125 I-labelled anti-IgE. RESULTS: Most IgE reactivity of the BPP fraction was due to the 5-7 kDa amino-terminal fragment of Ara h 1. This finding was confirmed by the use of the fragment in recombinant form, to which 25/55 of the sera was IgE-positive. CONCLUSION: The amino-terminal fragment of Ara h 1, a member of a family of small anti-microbial proteins, is an allergen independent of the carboxy-terminal fragment of Ara h 1.

The Role of Religious Socialization and Religiosity in African American and Caribbean Black Adolescents' Sexual Initiation.

This study determined the nature of the associations between religious socialization, religiosity, and adolescent sexual initiation. Data originated from the National Survey of American Life-Adolescent (n = 1170), a nationally representative study of black adolescents. Factor analysis, structural equation modeling, and logistic regression were used to evaluate hypotheses. Results indicated that as black adolescents received more messages about religious beliefs and practices, their religiosity was greater and, in turn, they were less likely to report sexual initiation; findings varied by ethnicity, gender, and age. Findings contribute to understanding religious socialization and its association with sexual initiation.

Seeds of HOPE: Incorporating Community-Based Strategies to Implement a Weight-Loss and Empowerment Intervention in Eastern North Carolina.

Seeds of HOPE (Health, Opportunities, Partnerships, and Empowerment) was a community-based participatory research project designed to implement an evidence-based weight loss and empowerment intervention aimed at increasing self-efficacy through peer support. HOPE Works, its model, has yielded significant weight loss and increases in participants' ability to set and achieve goals, including in consumption of fruits and vegetables and in physical activity.METHODS The Seeds of HOPE project was implemented through 3 peer leader recruitment approaches using Circle Leaders. Study participants met in hope circles to discuss various topics aimed at developing healthier lifestyles. Changes at 6-month intervals were assessed in health behaviors, weight, and hope. One-way ANOVA was used to test differences in mean change in weight and hope across the 3 recruitment approaches.RESULTS Study participants (N = 146) lost weight (-3.3 pounds, P = 0.003, 95% CI: -5.40 - -1.27). Fruit consumption significantly increased (P = 0.04, 95% CI: 0.01-0.41) while no changes in vegetable consumption or physical activity were observed. No differences based on different leader recruitment approaches were found.LIMITATIONS There were limitations in the power to fully detect differences across Circle Leader recruitment approaches; loss-to-follow up and representativeness of the sample were present. Further research is needed to determine if these approaches would be successful in other rural counties, with different genders, or with individuals in urban and/or higher socioeconomic populations.CONCLUSIONS Participants found modest success in weight loss and fruit consumption. Authors find that community engagement strategies are helpful in finding mutually acceptable adaptations to implement interventions such as Seeds of HOPE in diverse communities.

Iron incorporation into MnSOD A (bacterial Mn-dependent superoxide dismutase) leads to the formation of a peroxidase/catalase implicated in oxidative damage to bacteria.

BACKGROUND: Mn/Fe-superoxide dismutase (SOD) is a family of enzymes essential for organisms to be able to cope with oxygen. These enzymes bound to their classical metals catalyze the dismutation of the free radical superoxide anion (O2(-)) to H2O2 and molecular oxygen. E. coli has the manganese-dependent SOD A and the iron-dependent SOD B. METHODS: Strains of E. coli overexpressing SOD A or SOD B were grown in media with different metal compositions. SODs were purified and their metal content and SOD activity were determined. Those proteins were incubated with H2O2 and assayed for oxidation of Amplex red or o-phenylenediamine, consumption of H2O2, release of iron and protein radical formation. Cell survival was determined in bacteria with MnSOD A or FeSOD A after being challenged with H2O2. RESULTS: We show for the first time that the bacterial manganese-dependent SOD A when bound to iron (FeSOD A) has peroxidase activity. The in vivo formation of the peroxidase FeSOD A was increased when media had higher levels of iron because of a decreased manganese metal incorporation. In comparison to bacteria with MnSOD A, cells with FeSOD A had a higher loss of viability when exposed to H2O2. GENERAL SIGNIFICANCE: The biological occurrence of this fundamental antioxidant enzyme in an alternative iron-dependent state represents an important source of free radical formation.

Client history and violence on direct care workers in the home care setting.

BACKGROUND: Health care workers providing home care are frequently unaware of their client's history of violence or mental illness/substance abuse disorder, recognized risk factors for workplace violence. This study estimated the associations between these factors and experiencing client violence among direct care workers in the home settings (DCWHs). METHODS: Acts and threats of violence were estimated using data from an anonymous survey among DCWHs (n = 876) working at two large home care agencies. Logistic regressions were performed to produce odds ratios. RESULTS: Physical acts and physical or verbal threats of client violence were associated with providing homecare to clients with a violence history (adjusted ORs = 6.60 and 10.78, respectively), whereas threats of client violence (adjusted OR = 5.80) were associated with caring for clients with a mental illness/substance abuse disorder. CONCLUSIONS: Policy and practices that support the communication of appropriate client risk information may reduce the likelihood of workplace violence among DCWHs. Am. J. Ind. Med. 59:1130-1135, 2016. © 2016 Wiley Periodicals, Inc.

Analysis of glutathione S-transferase allergen cross-reactivity in a North American population: Relevance for molecular diagnosis.

BACKGROUND: It is not clear whether cross-reactivity or cosensitization to glutathione S-transferases (GSTs) occurs in tropical and subtropical environments. In the United States, Bla g 5 is the most important GST allergen and lack of coexposure to GSTs from certain species allows a better assessment of cross-reactivity. OBJECTIVES: To examine the molecular structure of GST allergens from cockroach (Bla g 5), dust mites (Der p 8 and Blo t 8), and helminth (Asc s 13) for potential cross-reactive sites, and to assess the IgE cross-reactivity of sensitized patients from a temperate climate for these allergens for molecular diagnostic purposes. METHODS: Four crystal structures were determined. Sera from patients allergic to cockroach and mite were tested for IgE reactivity to these GSTs. A panel of 6 murine anti-Bla g 5 mAb was assessed for cross-reactivity with the other 3 GSTs using antibody binding assays. RESULTS: Comparisons of the allergen structures, formed by 2-domain monomers that dimerize, revealed few contiguous regions of similar exposed residues, rendering cross-reactivity unlikely. Accordingly, anti-Bla g 5 or anti-Der p 8 IgE from North American patients did not recognize Der p 8 or Bla g 5, respectively, and neither showed binding to Blo t 8 or Asc s 13. A weaker binding of anti-Bla g 5 IgE to Der p 8 versus Bla g 5 (∼ 100-fold) was observed by inhibition assays, similar to a weak recognition of Der p 8 by anti-Bla g 5 mAb. Patients from tropical Colombia had IgE to all 4 GSTs. CONCLUSIONS: The lack of significant IgE cross-reactivity among the 4 GSTs is in agreement with the low shared amino acid identity at the molecular surface. Each GST is needed for accurate molecular diagnosis in different geographic areas.

Primary identification, biochemical characterization, and immunologic properties of the allergenic pollen cyclophilin cat R 1.

Cyclophilin (Cyp) allergens are considered pan-allergens due to frequently reported cross-reactivity. In addition to well studied fungal Cyps, a number of plant Cyps were identified as allergens (e.g. Bet v 7 from birch pollen, Cat r 1 from periwinkle pollen). However, there are conflicting data regarding their antigenic/allergenic cross-reactivity, with no plant Cyp allergen structures available for comparison. Because amino acid residues are fairly conserved between plant and fungal Cyps, it is particularly interesting to check whether they can cross-react. Cat r 1 was identified by immunoblotting using allergic patients' sera followed by N-terminal sequencing. Cat r 1 (∼ 91% sequence identity to Bet v 7) was cloned from a cDNA library and expressed in Escherichia coli. Recombinant Cat r 1 was utilized to confirm peptidyl-prolyl cis-trans-isomerase (PPIase) activity by a PPIase assay and the allergenic property by an IgE-specific immunoblotting and rat basophil leukemia cell (RBL-SX38) mediator release assay. Inhibition-ELISA showed cross-reactive binding of serum IgE from Cat r 1-allergic individuals to fungal allergenic Cyps Asp f 11 and Mala s 6. The molecular structure of Cat r 1 was determined by NMR spectroscopy. The antigenic surface was examined in relation to its plant, animal, and fungal homologues. The structure revealed a typical cyclophilin fold consisting of a compact ß-barrel made up of seven anti-parallel ß-strands along with two surrounding α-helices. This is the first structure of an allergenic plant Cyp revealing high conservation of the antigenic surface particularly near the PPIase active site, which supports the pronounced cross-reactivity among Cyps from various sources.

Community Member and Stakeholder Perspectives on a Healthy Environment Initiative in North Carolina.

INTRODUCTION: The North Carolina Community Transformation Grant Project (NC-CTG) aimed to implement policy, system, and environmental strategies to promote healthy eating, active living, tobacco-free living, and clinical and community preventive services to advance health equity and reduce health disparities for the state's most vulnerable communities. This article presents findings from the Health Equity Collaborative Evaluation and Implementation Project, which assessed community and stakeholder perceptions of health equity for 3 NC-CTG strategies: farmers markets, shared use, and smoke-free multiunit housing. METHODS: In a triangulated qualitative evaluation, 6 photo elicitation (PE) sessions among 45 community members in 1 urban and 3 rural counties and key informant interviews among 22 stakeholders were conducted. Nine participants from the PE sessions and key informant interviews in the urban county subsequently participated in a stakeholder power analysis and mapping session (SPA) to discuss and identify people and organizations in their community perceived to be influential in addressing health equity-related issues. RESULTS: Evaluations of the PE sessions and key informant interviews indicated that access (convenience, cost, safety, and awareness of products and services) and community fit (community-defined quality, safety, values, and norms) were important constructs across the strategies. The SPA identified specific community- and faith-based organizations, health care organizations, and local government agencies as key stakeholders for future efforts. CONCLUSIONS: Both community fit and access are essential constructs for promoting health equity. Findings demonstrate the feasibility of and need for formative research that engages community members and local stakeholders to shape context-specific, culturally relevant health promotion strategies.

Evaluating Policy, Systems, and Environmental Change Interventions: Lessons Learned From CDC's Prevention Research Centers.

INTRODUCTION: The field of public health is increasingly implementing initiatives intended to make policies, systems, and environments (PSEs) more supportive of healthy behaviors, even though the evidence for many of these strategies is only emerging. Our objective was 3-fold: 1) to describe evaluations of PSE-change programs in which the evaluators followed the steps of the Centers for Disease Control and Prevention's (CDC's) Framework for Program Evaluation in Public Health, 2) to share the resulting lessons learned, and 3) to assist future evaluators of PSE-change programs with their evaluation design decisions. METHODS: Seven Prevention Research Centers (PRCs) applied CDC's framework to evaluate their own PSE-change initiatives. The PRCs followed each step of the framework: 1) engage stakeholders, 2) describe program, 3) focus evaluation design, 4) gather credible evidence, 5) justify conclusions, and 6) ensure use and share lessons learned. RESULTS: Evaluation stakeholders represented a range of sectors, including public health departments, partner organizations, and community members. Public health departments were the primary stakeholders for 4 of the 7 evaluations. Four PRCs used logic models to describe the initiatives being evaluated. Their evaluations typically included both process and outcome questions and used mixed methods. Evaluation findings most commonly focused on contextual factors influencing change (process) and the adoption or implementation of PSE-change strategies (outcome). Evaluators shared lessons learned through various channels to reach local stakeholders and broader public health audiences. CONCLUSION: Framework for Program Evaluation in Public Health is applicable to evaluations of PSE-change initiatives. Using this framework to guide such evaluations builds practice-based evidence for strategies that are increasingly being used to promote healthful behaviors.

Occupational Health Providers' Perceptions of Employee Vaccine Hesitancy.

OBJECTIVE: This study examined the perspectives of occupational health providers (OHPs) on the most frequently encountered clinically relevant reasons for employee vaccine hesitancy. METHODS: We conducted an anonymous, online, cross-sectional survey of US OHPs ( N = 217). The survey asked OHPs about the major reasons that employees cite for being unwilling to receive the following three categories of vaccines: COVID-19, annual influenza, and others relevant to the workplace. RESULTS: Concern about adverse effects was the most frequently reported reason for employee vaccine hesitancy for each vaccine category. Mistrust was reported more frequently for COVID-19 than for the influenza vaccine or other vaccines (χ 2P < 0.05). Targets of employee mistrust included government and researchers or scientists, but mistrust of healthcare providers was uncommon. CONCLUSIONS: These results can be used to inform interventions to address vaccine hesitancy in the occupational health setting.

What "price" means when buying food: insights from a multisite qualitative study with Black Americans.

OBJECTIVES: We explored the role of price in the food purchasing patterns of Black adults and youths. METHODS: We analyzed qualitative data from interviews and focus groups with socioeconomically diverse, primarily female, Black adults or parents (n = 75) and youths (n = 42) in 4 US cities. Interview protocols were locality specific, but all were designed to elicit broad discussion of food marketing variables. We performed a conventional qualitative content analysis by coding and analyzing data from each site to identify common salient themes. RESULTS: Price emerged as a primary influence on food purchases across all sites. Other value considerations (e.g., convenience, food quality, healthfulness of product, and family preferences) were discussed, providing a more complex picture of how participants considered the price of a product. CONCLUSIONS: Food pricing strategies that encourage consumption of healthful foods may have high relevance for Black persons across income or education levels. Accounting for how price intersects with other value considerations may improve the effectiveness of these strategies.

Perspectives on Benefits and Challenges to Developing a Co-Teaching and Co-Learning Exchange between Community and Academic Partners.

BACKGROUND: To train future professionals in health disparities and social determinants of health, academic health centers often use curricula exclusively developed and instructed by faculty. OBJECTIVE: To examine the perceptions and attitudes of faculty and community stakeholders towards the benefits of and challenges to developing co-teaching/co-learning exchange programs. METHODS: Faculty from six academic professional schools at a single institution and community members participated in focus groups. Interviews were video-recorded and reviewed for themes. RESULTS: Both faculty and community participants felt that partnering in the design and implementation of lectures addressing the social determinants of health could enhance curriculum and provide real-world context for the learning experience. CONCLUSIONS: Our findings add to the literature examining the benefits and challenges of engagement between faculty and community and offer new insights on the value of co-teaching/co-learning experiences.