Genetic Study of Elf5 and Ehf in the Mouse Salivary Gland.

Salivary gland (SG) development, maturation, and homeostasis require coordinated roles of transcription factors (TFs) that dictate specific cell identities and fate. The ETS family of proteins are important transcriptional drivers of diverse cell lineages, tissue development, and differentiation programs and hence are also likely to play an important role in the SG. Here we have leveraged genomic and epigenomic data of the SG to examine the expression profile of ETS genes and identified 2 closely related paralogs, Elf5 and Ehf, that are highly expressed in distinct epithelial subpopulations. By using a well-defined mouse knockout model of Elf5, we show that Elf5, despite its enriched expression in the acinar cells, is functionally dispensable for maintaining the homeostatic state of the adult SG epithelium. The lack of a discernible phenotype of the Elf5-null SG might be due to possible functional redundancy with Ehf or other ETS factors. To probe this possibility and to examine the specific consequences of Ehf loss in the SG, we used CRISPR-Cas9 to generate mice in which the DNA-binding ETS domain of Ehf is disrupted due to an insertion mutation. We demonstrate that the Ehf mutant (EhfMut) mice exhibit a distinct cellular phenotype with decreased granular convoluted tubules that are accompanied by an increased accumulation of the intercalated Sox9-positive ductal cell population. Interestingly, the ductal phenotype of the EhfMut animals is highly pronounced in males, reaffirming the established sexual dimorphism of the SG that exists in rodents. Our results show that unlike Elf5, Ehf plays a nonredundant role in directing ductal cell differentiation of the SG and highlights the phenotypic subtlety in mutant mice of closely related TFs and the importance of careful consideration of cell type-specific studies.

Savoring Interventions Increase Positive Emotions After a Social-Evaluative Hassle.

Achieving a high quality of life is dependent upon how individuals face adversity. Positive psychological interventions are well-suited to support coping efforts; however, experimental research is limited. The purpose of the current research was to examine whether different savoring interventions could increase important coping resources (i.e., positive emotions) in response to a social-evaluative hassle. We completed an experimental mixed subject design study with a university student sample. All participants completed a hassle induction task and were then randomly assigned into different intervention groups. Positive emotion ratings were collected at three points in time (baseline, post-induction task, post-intervention). Results revealed a significant time x intervention interaction effect, such that individuals in the savoring the moment intervention reported higher levels of positive emotions (at post-intervention) compared to individuals assigned to the true control group, guided imagery control group, and savoring through reminiscence intervention. Such findings represent a significant extension to savoring theory and offer support for savoring the moment exercises as a primary prevention strategy to bolster effective responses to social-evaluative hassles.

A qualitative analysis of pharmacists' responses to naloxone inquiries in rural Georgia.

Given their critical gatekeeper function regarding naloxone access in rural areas, the purpose of the current study was to examine rural pharmacists' knowledge and attitudes regarding naloxone use and access (Georgia, 2019), all 364 rural community pharmacies in Georgia were contacted using "secret shopper" methodology and asked about the availability, accessibility, and use of naloxone. A script provided specific questions for data collection. Callers recorded general notes about the call, including the dispositions and impressions of the pharmacists. Thematic analysis was conducted to identify key themes. Rural pharmacists exhibited two classes of attributes: Active Facilitators and Passive Hindrances. Active facilitators supported callers in their search for naloxone, but passive hindrances discouraged callers in a way that was not perceived to be malicious or intentional. Three types of active facilitators emerged: positive approach, helpful suggestions, and knowledgeable and informed. Two types of passive hindrances emerged: negative attitudes and insufficient knowledge. Community pharmacies are sometimes the only formal healthcare access point in rural communities. It is vital for pharmacists to be active facilitators of naloxone access for maximized public health impact. Specific interventions are needed to increase the ability of rural pharmacists to support access to naloxone.

Disparities Elimination Summer Research Experience (DESRE): An Intensive Summer Research Training Program to Promote Diversity in the Biomedical Research Workforce.

The Disparities Elimination Summer Research Experience (DESRE) was created to provide hands-on health equity research training opportunities to undergraduate and graduate students, particularly those from backgrounds underrepresented in biomedical research. Funded by NIH's National Institute on Minority Health and Health Disparities, a total of 22 students participated in 4 annual cycles of an intensive, 6-week, full-time, residential research training program consisting of didactics, community immersion experiences, peer mentoring, ethics training, and hands-on health disparities research. Demand for the program was high; by the 4th year of implementation, more than 500 applications were received for the cohort's six slots. More than half of DESRE participants came from minority-serving institutions and/or identified as a member of a minority group. Students reported a significant increase in self-reported competency across all of the program's 26 learning objectives from pre- to post-assessment. Further, the program had a 77% success rate in promoting a career in biomedical research and/or health disparities elimination, including 100% of minority participants either entering a graduate program and/or entering careers focused on health equity. Key success factors and lessons learned are discussed.

Transcriptomic and Single-Cell Analysis of the Murine Parotid Gland.

The salivary complex of mammals consists of 3 major pairs of glands: the parotid, submandibular, and sublingual glands. While the 3 glands share similar functional properties, such as saliva secretion, their differences are largely based on the types of secretions they produce. While recent studies have begun to shed light on the underlying molecular differences among the glands, few have examined the global transcriptional repertoire over various stages of gland maturation. To better elucidate the molecular nature of the parotid gland, we have performed RNA sequencing to generate comprehensive and global gene expression profiles of this gland at different stages of maturation. Our transcriptomic characterization and hierarchical clustering analysis with adult organ RNA sequencing data sets has identified a number of molecular players and pathways that are relevant for parotid gland biology. Moreover, our detailed analysis has revealed a unique parotid gland-specific gene signature that may represent important players that could impart parotid gland-specific biological properties. To complement our transcriptomic studies, we have performed single-cell RNA sequencing to map the transcriptomes of parotid epithelial cells. Interrogation of the single-cell transcriptomes revealed the degree of molecular and cellular heterogeneity of the various epithelial cell types within the parotid gland. Moreover, we uncovered a mixed-lineage population of cells that may reflect molecular priming of differentiation potentials. Overall our comprehensive studies provide a powerful tool for the discovery of novel molecular players important in parotid gland biology.

The Impact of Rurality on Likelihood of Drunk Driving and Riding With a Driver Under the Influence Among High School Students.

PURPOSE: The purpose of this study was to examine the importance of rural location in the likelihood of adolescent drunk driving and riding in a vehicle with a driver under the influence while controlling for a variety of student-, school-, and county-level factors. METHODS: Data from the 2013 Georgia Student Health Survey (GSHS) II (a statewide assessment of student health in public school students in Georgia) were analyzed using multilevel binary regressions to examine rural-urban differences in prevalence of driving under the influence (11th and 12th graders only; n = 114,907) and riding with a driver under the influence (9th -12th graders; n = 258,610), controlling for school-level race, gender, and income, in addition to county-level education level, unemployment, alcohol use, and smoking. FINDINGS: Across geographies, students were twice as likely to report riding with a driver under the influence (10.32%) as driving under the influence (4.16%). While both outcomes were more likely among rural adolescents in unadjusted analyses, in adjusted analyses, rural adolescents were not significantly more likely to drive under the influence (ORadj = 1.19; P = .055), but they were significantly more likely to ride with a driver under the influence (ORadj = 1.18; P = .002). CONCLUSIONS: Our findings suggest that riding with a driver under the influence rather than driving under the influence may be a more pressing public health issue for adolescents in rural areas. Future research focused on the formative work necessary to build novel, culturally tailored interventions should be conducted to minimize the associated substantial burden of motor vehicle deaths within rural adolescents.

Discrepancy in Motivation for Weight Loss and Exercise in Rural Patients.

OBJECTIVES: We explored the extent to which discrepancy between motivation for weight loss and exercise is related to obesity among rural patients with chronic disease, and identified the psychosocial correlates of this discrepancy. METHODS: 497 patients with diabetes and/or hypertension were recruited from a network of Federally Qualified Health Centers in the rural South and completed a battery of assessments. RESULTS: Most persons in the sample (83.1%) were overweight and 65.0% were obese. For motivation for change, 70.8% reported being in the Action stage or higher for weight loss, whereas only 24.9% reported being in the Action stage or higher for motivation for exercise. When controlling for age, education level, income, sex, and race/ethnicity, individuals who were motivated for weight loss but not exercise were twice as likely to be obese (p = .005). Race and sex were significantly associated with this discrepancy, with African-American patients 1.7 times as likely (p = .05) and women 2.3 times as likely to be discrepant (p = .001). CONCLUSIONS: Findings underscore the importance of focusing not simply on weight loss among rural patients with chronic disease, but rather to incorporate specific activities designed to build simultaneous motivation for engaging in exercise.

Recent Alcohol, Tobacco, and Substance Use Variations between Rural and Urban Middle and High School Students.

The use of addictive substances by adolescents is a major public health concern; however, rural vs. urban variations are poorly understood. The purpose of the current study was to examine rural-urban differences in the prevalence of recent use of 11 substances in grades 6-12 in a statewide sample of students from the Georgia Student Health Survey II (n = 513,909). We found that rural-urban differences in substance use depend largely upon grade level, with rural middle school students demonstrating higher rates of alcohol, smoking tobacco, and chewing tobacco use, and urban high school students demonstrating higher rates of illicit drugs.

Women's health care: the experiences and behaviors of rural and urban lesbians in the USA.

INTRODUCTION: Previous research has consistently demonstrated that, in comparison to their cisgender heterosexual counterparts, lesbians face a multitude of women's healthcare-related disparities. However, very little research has been conducted that takes an intersectionality approach to examining the potential influences of rural-urban location on the health-related needs and experiences of lesbians. The purpose of this study was to quantitatively compare rural and urban lesbians' access to women's health care, experiences with women's healthcare providers (WHCPs), and preventive behavior using a large, diverse sample of lesbians from across the USA. METHODS: A total of 895 (31.1% rural and 68.9% urban) lesbian-identified cisgender women (ie not transgender) from the USA participated in the current online study. As part of a larger parent study, participants were recruited from across the USA through email communication to lesbian, gay, bisexual, and transgender (LGBT)-focused organizations and online advertisements. Participants were asked to complete a series of questions related to their women's healthcare-related experiences and behaviors (ie access to care, experiences with WHCPs, and preventive behavior). A series of χ2 analyses were utilized in order to examine rural-urban differences across dependent variables. RESULTS: An examination of sexual risks revealed that relatively more rural lesbians reported at least one previous male sexual partner in comparison to the urban sample of lesbians (78.1% vs 69.1%, χ2(1, N=890)=7.56, p=0.006). A similarly low percentage of rural (42.4%) and urban (42.9%) lesbians reported that they have a WHCP that they see on a regular basis for preventive care. In terms of experiences with WHCP providers, relatively fewer rural lesbians indicated that their current WHCP had discussed/recommended the human papillomavirus (HPV) vaccination in comparison to urban lesbians (27.5% vs. 37.2%; χ2 (1, N=796)=7.24, p=0.007). No other rural-urban differences in experiences with WHCPs emerged - few rural and urban lesbians had been asked about their sexual orientation by their WHCP (38.8% and 45.0%, respectively), been provided with appropriate safe-sex education by their WHCP (21.4% and 25.3%), had their last HIV/sexually transmitted infection screening instigated by their WHCP (if applicable; 15.7% and 19.5%), and reported that their WHCP seems knowledgeable about lesbian health concerns (56.5% vs 54.6%). In terms of preventive behavior, significantly fewer rural lesbians aged 40 years or more had received a mammogram in the past 3 years (63.2% vs 83.2%; χ2(1, N=163)=8.36, p=0.004) when compared to their urban counterparts. No other significant rural-urban differences in preventive behaviors emerged. A similarly low percentage of rural and urban lesbians indicated that they have received the HPV vaccination (22.8% and 29.0%, respectively) and/or have had a HIV/STI screening (43.0% and 47.8%), Pap test (62.0% and 64.5%) or breast exam (59.2% and 62.8%), in the past 3 years. CONCLUSIONS: The current findings highlight that rural lesbians in the USA, in comparison to urban lesbians, may experience elevated health risks related to being more likely to have at least one previous male sexual partner, less likely to be recommended the HPV vaccination by a WHCP, and, for those 40 or older, less likely to receive routine mammogram screenings. Furthermore, rural lesbians appear to engage in similarly low rates of HPV vaccination and regular HIV/STI screenings, Pap tests, and clinical breast exams as their urban counterparts. Given the increased cervical and breast cancer risks associated with rural living, the current findings underscore the dire need for health promotion efforts aimed at increasing rural lesbians' engagement in routine pelvic and breast exams.

Discrepancy between Actual and Perceived Weight Status in Rural Patients: Variations by Race and Gender.

The study's goal was to determine if differences in weight misperception by race and/or gender occur within a sample of economically disadvantaged rural patients with diabetes and/or hypertension. Diabetic and hypertensive patients were enrolled in the study from a network of federally qualified health centers (FQHCs) in the rural South. Multivari-ate logistic regression analysis suggests that, even when controlling for age, education level, employment status, and poverty, rural African American patients with chronic disease are more likely than their White counterparts to misperceive their weight status (OR = 1.709, p = .037). This difference in perceived weight occurred despite the absence of an underlying difference in actual weight status between African American and White patients (p = .171). In addition, rural men were much more likely than rural women to misperceive their weight status (OR = 2.688, p < .001). Implications for intervention development and implementation are discussed.

Inhibition of proliferation and invasion in 2D and 3D models by 2-methoxyestradiol in human melanoma cells.

Despite the recent advances in the clinical management of melanoma, there remains a need for new pharmacological approaches to treat this cancer. 2-methoxyestradiol (2ME) is a metabolite of estrogen that has shown anti-tumor effects in many cancer types. In this study we show that 2ME treatment leads to growth inhibition in melanoma cells, an effect associated with entry into senescence, decreased pRb and Cyclin B1 expression, increased p21/Cip1 expression and G2/M cell cycle arrest. 2ME treatment also inhibits melanoma cell growth in colony formation assay, including cell lines with acquired resistance to BRAF and BRAF+MEK inhibitors. We further show that 2ME is effective against melanoma with different BRAF and NRAS mutational status. Moreover, 2ME induced the retraction of cytoplasmic projections in a 3D spheroid model and significantly decreased cell proliferation in a 3D skin reconstruct model. Together our studies bring new insights into the mechanism of action of 2ME allowing melanoma targeted therapy to be further refined. Continued progress in this area is expected to lead to improved anti-cancer treatments and the development of new and more effective clinical analogues.

Targeting the hedgehog transcription factors GLI1 and GLI2 restores sensitivity to vemurafenib-resistant human melanoma cells.

BRAF inhibitor (BRAFi) therapy for melanoma patients harboring the V600E mutation is initially highly effective, but almost all patients relapse within a few months. Understanding the molecular mechanisms underpinning BRAFi-based therapy is therefore an important issue. Here we identified a previously unsuspected mechanism of BRAFi resistance driven by elevated Hedgehog (Hh) pathway activation that is observed in a cohort of melanoma patients after vemurafenib treatment. Specifically, we demonstrate that melanoma cell lines, with acquired in vitro-induced vemurafenib resistance, show increased levels of glioma-associated oncogene homolog 1 and 2 (GLI1/GLI2) compared with naïve cells. We also observed these findings in clinical melanoma specimens. Moreover, the increased expression of the transcription factors GLI1/GLI2 was independent of canonical Hh signaling and was instead correlated with the noncanonical Hh pathway, involving TGFß/SMAD (transforming growth factor-ß/Sma- and Mad-related family) signaling. Knockdown of GLI1 and GLI2 restored sensitivity to vemurafenib-resistant cells, an effect associated with both growth arrest and senescence. Treatment of vemurafenib-resistant cells with the GLI1/GLI2 inhibitor Gant61 led to decreased invasion of the melanoma cells in a three-dimensional skin reconstruct model and was associated with a decrease in metalloproteinase (MMP2/MMP9) expression and microphthalmia transcription factor upregulation. Gant61 monotherapy did not alter the drug sensitivity of naïve cells, but could reverse the resistance of melanoma cells chronically treated with vemurafenib. We further noted that alternating dosing schedules of Gant61 and vemurafenib prevented the onset of BRAFi resistance, suggesting that this could be a potential therapeutic strategy for the prevention of therapeutic escape. Our results suggest that targeting the Hh pathway in BRAFi-resistant melanoma may represent a viable therapeutic strategy to restore vemurafenib sensitivity, reducing or even inhibiting the acquired chemoresistance in melanoma patients.

Connection between Depression and Inability to Fill Prescriptions in Rural FQHC Patients with Chronic Disease.

The purpose of this paper was to (1) examine the rates of elevated depression symptoms among a sample of rural Federally Qualified Health Center (FQHC) patients with chronic disease and (2) determine if an inability to afford general prescription medications within the past 12 months is a significant predictor of depression symptoms among these patients. These data came from Project EDUCATE, an ongoing five-year study designed to be a large-scale, multifocal examination of the needs and experiences of rural FQHC patients with hypertension and/or diabetes. A total of 497 rural FQHC patients completed surveys (including a series of psychosocial questions, the Multigroup Ethnic Identity Measure and the Center for Epidemiologic Studies Depression scale) as part of phase one of the project; 438 of these with complete data are included in the current analytic sample. Results revealed that 53.0% of the sample screened positive for depression, and over half of those who screened positive reported not being able to afford their prescription medications at least once within the past 12 months (51.3% vs. 26.3% non-depressed). Further, even after controlling for age, ethnic identity attachment, sex, education level, employment status, income, insurance status, recent inability to afford needed medical care, hypertensive status, mental health diagnosis, and family history of mental illness, patients who could not afford to fill their prescriptions in the past 12 months were 2.6 times as likely to screen positive for depression (ORADJ = 2.476, p = 0.002) as those who could afford their medications. Overall, results of this study suggest that, among rural patients diagnosed with chronic disease, depressive symptomatology may be alarmingly high and an inability to afford medications may be an important risk factor for depression symptoms. These results highlight the need for increased attention to prescription medication affordability among rural patients with chronic disease in order to reduce the risk of comorbid depression.

Ethnic Identity Attachment and Motivation for Weight Loss and Exercise Among Rural, Overweight, African-American Women.

Rural and minority women are disproportionately impacted by the obesity epidemic; however, little research has studied the intersection of these disparity groups. The purpose of this study was to examine the influence of racial identity on motivation for weight loss and exercise among rural, African-American women with an obesity-linked chronic disease. A total of 154 African-American women were recruited from the patient population of a Federally Qualified Health Center in the rural South to complete a questionnaire battery including the Multigroup Ethnic Identity Measure and separate assessments of motivation for weight loss and exercise. Multivariate analyses, controlling for age, education status, insurance status, and body mass index revealed that attachment to ethnic identity was predictive of motivation for exercise but not for weight loss. Our findings suggest that attachment to ethnic identity may be an important factor in motivation for change among African-American women, particularly with respect to exercise, with direct implications for the development of culturally and geographically tailored weight loss interventions.

Differences in Psychosocial Predictors of Obesity Among LGBT Subgroups.

PURPOSE: The purpose of the current study was to examine the overall presence of and differences in rates of overweight/obesity among a large, nationally diverse sample of lesbian, gay, bisexual, transgender (LGBT)-identified individuals (i.e., cisgender lesbians, cisgender gay men, cisgender bisexual women, cisgender bisexual men, transgender women, and transgender men) and to identify specific psychosocial predictors of obesity within each of the six LGBT subgroups. METHODS: A total of 2702 LGBT-identified participants participated in the online study. Participants completed a series of demographic questions (including weight and height) and the Depression Anxiety Stress Scale 21. RESULTS: The percentage of participants who were overweight/obese did not differ significantly across LGBT subgroups, with 61.1% of the total sample being overweight/obese. However, the percentage of participants who self-reported body mass indexes in the obese range differed significantly across the six LGBT subgroups, with the highest prevalence in transgender men (46.0%). In addition, the predictors of obesity varied by subgroup, with age a significant predictor for cisgender lesbians, cisgender gay men, and cisgender bisexual women, relationship status for cisgender bisexual women, employment status for both cisgender gay men and cisgender bisexual women, education level for cisgender lesbians, and depression, anxiety, and stress for cisgender gay men. None of the examined psychosocial factors emerged as predictors of obesity for cisgender bisexual men, transgender women, or transgender men. CONCLUSION: These findings suggest that there are substantial variations in the presence and predictors of obesity across LGBT subgroups that support the need for culturally tailored healthy weight promotion efforts within the LGBT community.

ΔNp63 regulates IL-33 and IL-31 signaling in atopic dermatitis.

Atopic dermatitis (AD) is the most common inflammatory skin disease with no well-delineated cause or effective cure. Here we show that the p53 family member p63, specifically the ΔNp63, isoform has a key role in driving keratinocyte activation in AD. We find that overexpression of ΔNp63 in transgenic mouse epidermis results in a severe skin phenotype that shares many of the key clinical, histological and molecular features associated with human AD. This includes pruritus, epidermal hyperplasia, aberrant keratinocyte differentiation, enhanced expression of selected cytokines and chemokines and the infiltration of large numbers of inflammatory cells including type 2 T-helper cells - features that are highly representative of AD dermatopathology. We further demonstrate several of these mediators to be direct transcriptional targets of ΔNp63 in keratinocytes. Of particular significance are two p63 target genes, IL-31 and IL-33, both of which are key players in the signaling pathways implicated in AD. Importantly, we find these observations to be in good agreement with elevated levels of ΔNp63 in skin lesions of human patients with AD. Our studies reveal an important role for ΔNp63 in the pathogenesis of AD and offer new insights into its etiology and possible therapeutic targets.

Essential role of HDAC6 in the regulation of PD-L1 in melanoma.

Histone deacetylases (HDACs), originally described as histone modifiers, have more recently been demonstrated to target a variety of other proteins unrelated to the chromatin environment. In this context, our present work demonstrates that the pharmacological or genetic abrogation of HDAC6 in primary melanoma samples and cell lines, down-regulates the expression of PD-L1, an important co-stimulatory molecule expressed in cancer cells, which activates the inhibitory regulatory pathway PD-1 in T-cells. Our data suggests that this novel mechanism of PD-L1 regulation is mainly mediated by the influence of HDAC6 over the recruitment and activation of STAT3. Additionally, we observed that selective HDAC6 inhibitors impairs tumor growth and reduce the in vivo expression of several inhibitory check-point molecules and other regulatory pathways involved in immunosurveillance. Most importantly, these results provide a key pre-clinical rationale and justification to further study isotype selective HDAC6 inhibitors as potential immuno-modulatory agents in cancer.

Differences in health risk behaviors across understudied LGBT subgroups.

OBJECTIVE: The purpose of the current study was to conduct a large-scale, geographically diverse comparison of health risk behaviors between a broad range of sexual orientation and gender identity groups to more fully understand the health risks of subgroups within the LGBT community. METHOD: A total of 3,279 individuals self-identifying as LGBT (e.g., lesbian, gay, bisexual, pansexual, transgender, genderqueer, and/or another gender or sexual minority) were recruited from across the United States through 2 sequentially implemented online recruitment methods. Participants completed a demographic assessment and the Health Risk Questionnaire (assessing diet and exercise, substance use and smoking, motor vehicle risks, sexual behaviors, violence, and medical risk-taking). RESULTS: Significant differences were found across genders for 18 of the 28 health risk behaviors investigated and across sexual orientations for 23 behaviors. Major differences emerged particularly with relation to diet and exercise behaviors, as well as sexual risk-taking, substance use, and medical risk-taking. Groups with notably elevated health risk behaviors included transgender women (diet and exercise behaviors), cisgender men (alcohol-related risk-taking), bisexual participants (substance use), and both transgender men and pansexual participants (self-harm). Differences between transgender participants and genderqueer or nonbinary participants were stark, indicating that these frequently combined groups have distinct health risk profiles. CONCLUSIONS: Results suggest that there are extensive and largely variable levels of engagement in health risk behaviors within the LGBT community. In addition, gender and sexual orientation subcategories that are traditionally collapsed into 1 category (i.e., transgender and bisexual) evidenced strikingly different risks when examined independently. Recommendations for future research and LGBT health promotion efforts are discussed.

Rural-Urban Differences in Perceptions of Child Overweight Among Children and Adolescents, Their Guardians and Health Care Professionals in the United States.

RESEARCH OBJECTIVE: Children and adolescents residing in rural environments with higher prevalence of an overweight population may develop inaccurate perceptions of a healthy weight. This study examines rural-urban differences in perceptions of child overweight among overweight (85 ≤ BMI percentile < 95) and obese children (BMI percentile ≥ 95), their guardians and health care providers (HCPs), and children's concomitant weight control. METHODS: The cross-sectional study was based on the 2005-2010 NHANES data (1,844 overweight and obese children and adolescents, aged 8-15 years). Rurality was defined using the 2003 RUCC. The weight status was based on the standardized measures of children's height and weight. Children reported whether they considered themselves overweight and whether they were trying to lose weight. Proxy respondents (ie, guardians) reported whether they considered their child to be overweight and whether an HCP had ever told them their child was overweight. Weighted percentages and predicted probabilities from multivariable logistic regressions were calculated, accounting for the complex, multistage, probability sampling design and nonresponse. FINDINGS: Rural residents comprised 18.8% of the study population; 41.8% of them were overweight and 58.2% were obese compared to 46.7% and 53.3% of urban peers, respectively. Misperceptions of children's weight status were 11.3 and 6.0 percentage points higher in rural children and their guardians, respectively. Recall of an HCP identification of child overweight was 6.3 percentage points lower among rural versus urban guardians. CONCLUSION: Obesity prevention efforts may be fostered by improving accuracy of child overweight perceptions. This may be particularly impactful in rural settings, where weight misperceptions are high.