Disruption of retinal inflammation and the development of diabetic retinopathy in mice by a CD40-derived peptide or mutation of CD40 in Müller cells.

AIMS/HYPOTHESIS: CD40 expressed in Müller cells is a central driver of diabetic retinopathy. CD40 causes phospholipase Cγ1 (PLCγ1)-dependent ATP release in Müller cells followed by purinergic receptor (P2X7)-dependent production of proinflammatory cytokines in myeloid cells. In the diabetic retina, CD40 and P2X7 upregulate a broad range of inflammatory molecules that promote development of diabetic retinopathy. The molecular event downstream of CD40 that activates the PLCγ1-ATP-P2X7-proinflammatory cytokine cascade and promotes development of diabetic retinopathy is unknown. We hypothesise that disruption of the CD40-driven molecular events that trigger this cascade prevents/treats diabetic retinopathy in mice. METHODS: B6 and transgenic mice with Müller cell-restricted expression of wild-type (WT) CD40 or CD40 with mutations in TNF receptor-associated factor (TRAF) binding sites were made diabetic using streptozotocin. Leucostasis was assessed using FITC-conjugated concanavalin A. Histopathology was examined in the retinal vasculature. Expression of inflammatory molecules and phospho-Tyr783 PLCγ1 (p-PLCγ1) were assessed using real-time PCR, immunoblot and/or immunohistochemistry. Release of ATP and cytokines were measured by ATP bioluminescence and ELISA, respectively. RESULTS: Human Müller cells with CD40 ΔT2,3 (lacks TRAF2,3 binding sites) were unable to phosphorylate PLCγ1 and release ATP in response to CD40 ligation, and could not induce TNF-α/IL-1ß secretion in bystander myeloid cells. CD40-TRAF signalling acted via Src to induce PLCγ1 phosphorylation. Diabetic mice in which WT CD40 in Müller cells was replaced by CD40 ΔT2,3 failed to exhibit phosphorylation of PLCγ1 in these cells and upregulate P2X7 and TNF-α in microglia/macrophages. P2x7 (also known as P2rx7), Tnf-α (also known as Tnf), Il-1ß (also known as Il1b), Nos2, Icam-1 (also known as Icam1) and Ccl2 mRNA were not increased in these mice and the mice did not develop retinal leucostasis and capillary degeneration. Diabetic B6 mice treated intravitreally with a cell-permeable peptide that disrupts CD40-TRAF2,3 signalling did not exhibit either upregulation of P2X7 and inflammatory molecules in the retina or leucostasis. CONCLUSIONS/INTERPRETATION: CD40-TRAF2,3 signalling activated the CD40-PLCγ1-ATP-P2X7-proinflammatory cytokine pathway. Src functioned as a link between CD40-TRAF2,3 and PLCγ1. Replacing WT CD40 with CD40 ΔT2,3 impaired activation of PLCγ1 in Müller cells, upregulation of P2X7 in microglia/macrophages, upregulation of a broad range of inflammatory molecules in the diabetic retina and the development of diabetic retinopathy. Administration of a peptide that disrupts CD40-TRAF2,3 signalling reduced retinal expression of inflammatory molecules and reduced leucostasis in diabetic mice, supporting the therapeutic potential of pharmacological inhibition of CD40-TRAF2,3 in diabetic retinopathy.

Adaptation and study protocol of the evidence-based Make Better Choices (MBC2) multiple diet and activity change intervention for a rural Appalachian population.

BACKGROUND: Rural Appalachian residents experience among the highest prevalence of chronic disease, premature mortality, and decreased life expectancy in the nation. Addressing these growing inequities while avoiding duplicating existing programming necessitates the development of appropriate adaptations of evidence-based lifestyle interventions. Yet few published articles explicate how to accomplish such contextual and cultural adaptation. METHODS: In this paper, we describe the process of adapting the Make Better Choices 2 (MBC2) mHealth diet and activity randomized trial and the revised protocol for intervention implementation in rural Appalachia. Deploying the NIH's Cultural Framework on Health and Aaron's Adaptation framework, the iterative adaptation process included convening focus groups (N = 4, 38 participants), conducting key informant interviews (N = 16), verifying findings with our Community Advisory Board (N = 9), and deploying usability surveys (N = 8), wireframing (N = 8), and pilot testing (N = 9. This intense process resulted in a comprehensive revision of recruitment, retention, assessment, and intervention components. For the main trial, 350 participants will be randomized to receive either the multicomponent MBC2 diet and activity intervention or an active control condition (stress and sleep management). The main outcome is a composite score of four behavioral outcomes: two outcomes related to diet (increased fruits and vegetables and decreased saturated fat intake) and two related to activity (increased moderate vigorous physical activity [MVPA] and decreased time spent on sedentary activities). Secondary outcomes include change in biomarkers, including blood pressure, lipids, A1C, waist circumference, and BMI. DISCUSSION: Adaptation and implementation of evidence-based interventions is necessary to ensure efficacious contextually and culturally appropriate health services and programs, particularly for underserved and vulnerable populations. This article describes the development process of an adapted, community-embedded health intervention and the final protocol created to improve health behavior and, ultimately, advance health equity. TRIAL REGISTRATION: ClinicalTrials.gov Identifier NCT04309461. The trial was registered on 6/3/2020.

Practitioners Assess Achievements and Challenges of Nonfatal Injury Surveillance.

OBJECTIVE: Injury surveillance relies on data coded for administrative rather than epidemiological accuracy. The Centers for Disease Control and Prevention (CDC) established the 5-year Surveillance Quality Improvement (SQI) initiative to advance consensus and methodology for injury epidemiology reporting and analysis. Evaluation of the positive predictive value of the CDC's injury surveillance definitions based on International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) and International Classification of Diseases, Tenth Revision, Clinical Modification (ICD-10-CM) coding in designated injury categories comprised much of the SQI initiative's work. The goal of the current study is to identify achievements and challenges in SQI as articulated by experienced injury epidemiology practitioners who participated in the CDC-funded SQI initiative. DESIGN, SETTING, AND PARTICIPANTS: We conducted semistructured interviews with 12 representatives of state and federal public health agencies who had participated extensively in the SQI initiative. The interviews were transcribed and coded using NVivo qualitative analysis software. Initial coding of the data involved both in vivo coding (using the words of participants) and coding of a priori themes. MAIN OUTCOME MEASURES: Qualitative analysis identified 2 overarching themes, variability among states and observations on the science of injury surveillance. RESULTS: Within the 2 broad themes, the respondents provided valuable insights regarding access to medical records, case definition validation, unique contributions of medical record abstracting, variations in the practice of medical coding, and the potential for use of data from medical record reviews in other injury-related areas. CONCLUSIONS: The contributions of the SQI initiative have provided valuable insights into ICD-10-CM case definitions for national injury surveillance. Challenges remain with regard to data access and quality with ongoing reliance on administrative datasets for injury surveillance.

Celebrity Cancer on Twitter: Mapping a Novel Opportunity for Cancer Prevention.

Social media platforms have the potential to facilitate the dissemination of cancer prevention and control messages following celebrity cancer diagnoses. However, cancer communicators have yet to systematically leverage these naturally occurring interventions on social media as these events are difficult to identify as they are unfolding and little research has analyzed their effect on social media conversations. In this study, we add to the research by analyzing how a celebrity cancer announcement influenced Twitter conversations in terms of the volume of social media messages and the type of content. Over a 9-day period, during which actor Ben Stiller announced that he had been treated for prostate cancer, we collected 1.2 million Twitter messages about cancer. We conducted automated content analyses to identify how often common cancer sites (prostate, breast, colon, or lung) were discussed. Then, we used manual content analysis on a sample of messages to identify cancer continuum content (awareness, prevention, early detection, diagnosis, treatment, survivorship, and end of life). Chi-square analyses were implemented to evaluate changes in cancer site and cancer continuum content before and after the announcement. We found that messages related to prostate cancer increased significantly more than expected for 2 days following Stiller's announcement. However, the number of cancer messages that described other cancer locations either did not increase or did not increase by the same magnitude. In terms of message content, results showed larger than expected increases in diagnosis messages. These results suggest opportunities to shape social media conversations following celebrity cancer announcements and increase prevention and early detection messages.

Retweeting Risk Communication: The Role of Threat and Efficacy.

Social media platforms like Twitter and Facebook provide risk communicators with the opportunity to quickly reach their constituents at the time of an emerging infectious disease. On these platforms, messages gain exposure through message passing (called "sharing" on Facebook and "retweeting" on Twitter). This raises the question of how to optimize risk messages for diffusion across networks and, as a result, increase message exposure. In this study we add to this growing body of research by identifying message-level strategies to increase message passing during high-ambiguity events. In addition, we draw on the extended parallel process model to examine how threat and efficacy information influence the passing of Zika risk messages. In August 2016, we collected 1,409 Twitter messages about Zika sent by U.S. public health agencies' accounts. Using content analysis methods, we identified intrinsic message features and then analyzed the influence of those features, the account sending the message, the network surrounding the account, and the saliency of Zika as a topic, using negative binomial regression. The results suggest that severity and efficacy information increase how frequently messages get passed on to others. Drawing on the results of this study, previous research on message passing, and diffusion theories, we identify a framework for risk communication on social media. This framework includes four key variables that influence message passing and identifies a core set of message strategies, including message timing, to increase exposure to risk messages on social media during high-ambiguity events.

Social Media Messages in an Emerging Health Crisis: Tweeting Bird Flu.

Limited research has examined the messages produced about health-related crises on social media platforms and whether these messages contain content that would allow individuals to make sense of a crisis and respond effectively. This study uses the crisis and emergency risk communication (CERC) framework to evaluate the content of messages sent via Twitter during an emerging crisis. Using manual and computer-driven content analysis methods, the study analyzed 25,598 tweets about the H7N9 virus that were produced in April 2013. The study found that a large proportion of messages contained sensemaking information. However, few tweets contained efficacy information that would help individuals respond to the crisis appropriately. Implications and recommendations for practice and future study are discussed.

Constructing the uncertainty of due dates.

By its nature, the date that a baby is predicted to be born, or the due date, is uncertain. How women construct the uncertainty of their due dates may have implications for when and how women give birth. In the United States as many as 15% of births occur before 39 weeks because of elective inductions or cesarean sections, putting these babies at risk for increased medical problems after birth and later in life. This qualitative study employs a grounded theory approach to understand the decisions women make on how and when to give birth. Thirty-three women who were pregnant or had given birth within the past 2 years participated in key informant or small-group interviews. The results suggest that women interpret the uncertainty of their due dates as a reason to wait for birth and as a reason to start the process early; however, information about a baby's brain development in the final weeks of pregnancy may persuade women to remain pregnant longer. The uncertainties of due dates are analyzed using Babrow's problematic integration, which distinguishes between epistemological and ontological uncertainty. The results point to a third type of uncertainty, axiological uncertainty. Axiological uncertainty is rooted in the values and ethics of outcomes.

A Simplified Method for Isolation and Culture of Retinal Pigment Epithelial Cells from Adult Mice.

Retinal pigment epithelial cells (RPE) are critical for the proper function of the retina. RPE dysfunction is involved in the pathogenesis of important retinal diseases, such as age-related macular degeneration, retinitis pigmentosa, and diabetic retinopathy. We present a streamlined approach for the isolation of RPE from murine adult eyes. In contrast to previously reported methods, this approach enables the isolation and culture of highly pure RPE from adult mice. This simple and fast method does not require extensive technical skill and is achievable with basic scientific tools and reagents. Primary RPE are isolated from C57BL/6 background mice aged 3- to 14-weeks by enucleation of the eye followed by the removal of the anterior segment. Enzymatic trypsinization and centrifugation are used to dissociate and isolate the RPE from the eyecup. In conclusion, this approach offers a quick and effective protocol for the utilization of RPE in the study of retinal function and disease.

Advanced Glycation End Products Upregulate CD40 in Human Retinal Endothelial and Müller Cells: Relevance to Diabetic Retinopathy.

CD40 induces pro-inflammatory responses in endothelial and Müller cells and is required for the development of diabetic retinopathy (DR). CD40 is upregulated in these cells in patients with DR. CD40 upregulation is a central feature of CD40-driven inflammatory disorders. What drives CD40 upregulation in the diabetic retina remains unknown. We examined the role of advanced glycation end products (AGEs) in CD40 upregulation in endothelial cells and Müller cells. Human endothelial cells and Müller cells were incubated with unmodified or methylglyoxal (MGO)-modified fibronectin. CD40 expression was assessed by flow cytometry. The expression of ICAM-1 and CCL2 was examined by flow cytometry or ELISA after stimulation with CD154 (CD40 ligand). The expression of carboxymethyl lysine (CML), fibronectin, and laminin as well as CD40 in endothelial and Müller cells from patients with DR was examined by confocal microscopy. Fibronectin modified by MGO upregulated CD40 in endothelial and Müller cells. CD40 upregulation was functionally relevant. MGO-modified fibronectin enhanced CD154-driven upregulation of ICAM-1 and CCL2 in endothelial and Müller cells. Increased CD40 expression in endothelial and Müller cells from patients with DR was associated with increased CML expression in fibronectin and laminin. These findings identify AGEs as inducers of CD40 upregulation in endothelial and Müller cells and enhancers of CD40-dependent pro-inflammatory responses. CD40 upregulation in these cells is associated with higher CML expression in fibronectin and laminin in patients with DR. This study revealed that CD40 and AGEs, two important drivers of DR, are interconnected.

Feasibility and efficacy of a novel audiovisual tool to increase colorectal cancer screening among rural Appalachian Kentucky adults.

Introduction: Residents of Appalachian regions in Kentucky experience increased colorectal cancer (CRC) incidence and mortality. While population-based screening methods, such as fecal immunochemical tests (FITs), can reduce many screening barriers, written instructions to complete FIT can be challenging for some individuals. We developed a novel audiovisual tool ("talking card") to educate and motivate accurate FIT completion and assessed its feasibility, acceptability, and efficacy. Materials and methods: We collected data on the talking card via: (1) cross-sectional surveys exploring perceptions of images, messaging, and perceived utility; (2) follow-up focus groups centered on feasibility and acceptability; and (3) efficacy testing in community-based FIT distribution events, where we assessed FIT completion rate, number of positive vs. negative screens, demographic characteristics of participants, and primary drivers of FIT completion. Results: Across the three study phases, 692 individuals participated. Survey respondents positively identified with the card's sounds and images, found it highly acceptable, and reported high-to-very high self-efficacy and response efficacy for completing FIT, with nearly half noting greater likelihood to complete screening after using the tool. Focus group participants confirmed the acceptability of the individuals featured on the card. Nearly 75% of participants provided a FIT accurately completed it, with most indicating the talking card, either alone or combined with another strategy, helped with completion. Discussion: To reduce CRC screening disparities among Appalachian Kentuckians, population-based screening using contextually relevant implementation strategies must be used alongside clinic-based education. The talking card represents a novel and promising strategy to promote screening uptake in both clinical and community settings.

CD40 Upregulation in the Retina of Patients With Diabetic Retinopathy: Association With TRAF2/TRAF6 Upregulation and Inflammatory Molecule Expression.

Purpose: CD40 is upregulated in the retinas of diabetic mice, drives pro-inflammatory molecule expression, and promotes diabetic retinopathy. The role of CD40 in diabetic retinopathy in humans is unknown. Upregulation of CD40 and its downstream signaling molecules TNF receptor associated factors (TRAFs) is a key feature of CD40-driven inflammatory disorders. We examined the expression of CD40, TRAF2, and TRAF6 as well as pro-inflammatory molecules in retinas from patients with diabetic retinopathy. Methods: Posterior poles from patients with diabetic retinopathy and non-diabetic controls were stained with antibodies against von Willebrand factor (labels endothelial cells), cellular retinaldehyde-binding protein (CRALBP), or vimentin (both label Müller cells) plus antibodies against CD40, TRAF2, TRAF6, ICAM-1, CCL2, TNF-α, and/or phospho-Tyr783 phospholipase Cγ1 (PLCγ1). Sections were analyzed by confocal microscopy. Results: CD40 expression was increased in endothelial and Müller cells from patients with diabetic retinopathy. CD40 was co-expressed with ICAM-1 in endothelial cells and with CCL2 in Müller cells. TNF-α was detected in retinal cells from these patients, but these cells lacked endothelial/Müller cell markers. CD40 in Müller cells from patients with diabetic retinopathy co-expressed activated phospholipase Cγ1, a molecule that induces TNF-α expression in myeloid cells in mice. CD40 upregulation in endothelial cells and Müller cells from patients with diabetic retinopathy was accompanied by TRAF2 and TRAF6 upregulation. Conclusions: CD40, TRAF2, and TRAF6 are upregulated in patients with diabetic retinopathy. CD40 associates with expression of pro-inflammatory molecules. These findings suggest that CD40-TRAF signaling may promote pro-inflammatory responses in the retinas of patients with diabetic retinopathy.

Community engagement through social media: A promising low-cost strategy for rural recruitment?

PURPOSE: For the same reasons that rural telehealth has shown promise for enhancing the provision of care in underserved environments, social media recruitment may facilitate more inclusive research engagement in rural areas. However, little research has examined social media recruitment in the rural context, and few studies have evaluated the feasibility of using a free social media page to build a network of rural community members who may be interested in a research study. Here, we describe the rationale, process, and protocols of developing and implementing a social media approach to recruit rural residents to participate in an mHealth intervention. METHODS: Informed by extensive formative research, we created a study Facebook page emphasizing community engagement in an mHealth behavioral intervention. We distributed the page to local networks and regularly posted recruitment and community messages. We collected data on the reach of the Facebook page, interaction with our messages, and initiations of our study intake survey. FINDINGS: Over 21 weeks, our Facebook page gained 429 followers, and Facebook users interacted with our social media messages 3,080 times. Compared to messages that described desirable study features, messages that described community involvement resulted in higher levels of online interaction. Social media and other recruitment approaches resulted in 225 people initiating our in-take survey, 9 enrolling in our pilot study, and 26 placing their names on a waiting list. CONCLUSIONS: A standalone social media page highlighting community involvement shows promise for recruiting in rural areas.

#Ebola: Emergency Risk Messages on Social Media.

Public health threats require effective communication. Evaluating effectiveness during a situation that requires emergency risk communication is difficult, however, because these events require an immediate response and collecting data may be secondary to more immediate needs. In this article, we draw on research analyzing the effectiveness of social media messages during times of imminent threat and research analyzing the emergency risk communication conceptual model in order to propose a method for evaluating emergency risk communication on social media. We demonstrate this method by evaluating 2,915 messages sent by local, state, and federal public health officials during the 2014 Ebola outbreak in the United States. The results provide empirical support for emergency risk communication and identify message strategies that have the potential to increase exposure to official communication on social media during future public health threats.

Evaluation of automated pre-treatment and transit in-vivo dosimetry in radiotherapy using empirically determined parameters.

BACKGROUND AND PURPOSE: First reports on clinical use of commercially automated systems for Electronic Portal Imaging Device (EPID)-based dosimetry in radiotherapy showed the capability to detect important changes in patient setup, anatomy and external device position. For this study, results for more than 3000 patients, for both pre-treatment verification and in-vivo transit dosimetry were analyzed. MATERIALS AND METHODS: For all Volumetric Modulated Arc Therapy (VMAT) plans, pre-treatment quality assurance (QA) with EPID images was performed. In-vivo dosimetry using transit EPID images was analyzed, including causes and actions for failed fractions for all patients receiving photon treatment (2018-2019). In total 3136 and 32,632 fractions were analyzed with pre-treatment and transit images respectively. Parameters for gamma analysis were empirically determined, balancing the rate between detection of clinically relevant problems and the number of false positive results. RESULTS: Pre-treatment and in-vivo results depended on machine type. Causes for failed in-vivo analysis included deviations in patient positioning (32%) and anatomy change (28%). In addition, errors in planning, imaging, treatment delivery, simulation, breath hold and with immobilization devices were detected. Actions for failed fractions were mostly to repeat the measurement while taking extra care in positioning (54%) and to intensify imaging procedures (14%). Four percent initiated plan adjustments, showing the potential of the system as a basis for adaptive planning. CONCLUSIONS: EPID-based pre-treatment and in-vivo transit dosimetry using a commercially available automated system efficiently revealed a wide variety of deviations and showed potential to serve as a basis for adaptive planning.

Mapping of agronomic traits, disease resistance and malting quality in a wide cross of two-row barley cultivars.

Wide crosses between genetically diverged parents may reveal novel loci for crop improvement that are not apparent in crosses between elite cultivars. The landrace Chevallier was a noted malting barley first grown in 1820. To identify potentially novel alleles for agronomic traits, Chevallier was crossed with the modern malting cultivar NFC Tipple generating two genetically diverse recombinant inbred line populations. Genetic maps were produced using genotyping-by-sequencing and 384-SNP genotyping, and the populations were phenotyped for agronomic traits to allow the identification of quantitative trait loci (QTL). Within the semi-dwarf 1 (sdw1) region on chromosome 3H Chevallier conferred increased plant height and reduced tiller number, with QTL for these traits explaining 79.4% and 35.2% of the phenotypic variance observed, respectively. Chevallier was also associated with powdery mildew susceptibility, with a QTL on 1H accounting for up to 19.1% of the variance and resistance at this locus most likely resulting from an Mla variant from Tipple. Two novel QTL for physiological leaf spotting were identified on 3H and 7H, explaining up to 17.1% of the variance and with the Chevallier allele reducing symptom severity on 7H. Preliminary micromalting analysis was also undertaken to compare the malting characteristics of Chevallier and Tipple. Chevallier malt contained significantly lower levels of both α-amylase and wort ß-glucan than Tipple malt, however no significant differences were observed for the remaining malting parameters measured. This suggests that the most obvious improvements in barley since the introduction of Chevallier are for agronomic traits such as height, yield and lodging resistance rather than for malting characteristics. Overall, our results demonstrate that this wide cross between Chevallier and Tipple may provide a source of novel QTL for barley breeding.

Lung Cancer Messages on Twitter: Content Analysis and Evaluation.

PURPOSE: The aim of this project was to describe and evaluate the levels of lung cancer communication across the cancer prevention and control continuum for content posted to Twitter during a 10-day period (September 30 to October 9) in 2016. METHODS: Descriptive and inferential statistics were used to identify relationships between tweet characteristics in lung cancer communication on Twitter and user-level data. Overall, 3,000 tweets published between September 30 and October 9 were assessed by a team of three coders. Lung cancer-specific tweets by user type (individuals, media, and organizations) were examined to identify content and structural message features. The study also assessed differences by user type in the use of hashtags, directed messages, health topic focus, and lung cancer-specific focus across the cancer control continuum. RESULTS: Across the universe of lung cancer tweets, the majority of tweets focused on treatment and the use of pharmaceutical and research interventions, followed by awareness and prevention and risk topics. Among all lung cancer tweets, messages were most consistently tweeted by individual users, and personal behavioral mobilizing cues to action were rare. CONCLUSIONS: Lung cancer advocates, as well as patient and medical advocacy organizations, with an interest in expanding the reach and effectiveness of social media efforts should monitor the topical nature of public tweets across the cancer continuum and consider integrating cues to action as a strategy to increase engagement and behavioral activation pertaining to lung cancer reduction efforts.