Face-valid phenotypes in a mouse model of the most common mutation in EEF1A2-related neurodevelopmental disorder.

De novo heterozygous missense mutations in EEF1A2, encoding neuromuscular translation-elongation factor eEF1A2, are associated with developmental and epileptic encephalopathies. We used CRISPR/Cas9 to recapitulate the most common mutation, E122K, in mice. Although E122K heterozygotes were not observed to have convulsive seizures, they exhibited frequent electrographic seizures and EEG abnormalities, transient early motor deficits and growth defects. Both E122K homozygotes and Eef1a2-null mice developed progressive motor abnormalities, with E122K homozygotes reaching humane endpoints by P31. The null phenotype is driven by progressive spinal neurodegeneration; however, no signs of neurodegeneration were observed in E122K homozygotes. The E122K protein was relatively stable in neurons yet highly unstable in skeletal myocytes, suggesting that the E122K/E122K phenotype is instead driven by loss of function in muscle. Nevertheless, motor abnormalities emerged far earlier in E122K homozygotes than in nulls, suggesting a toxic gain of function and/or a possible dominant-negative effect. This mouse model represents the first animal model of an EEF1A2 missense mutation with face-valid phenotypes and has provided mechanistic insights needed to inform rational treatment design.

Endogenous epitope tagging of eEF1A2 in mice reveals early embryonic expression of eEF1A2 and subcellular compartmentalisation of neuronal eEF1A1 and eEF1A2.

All vertebrate species express two independently-encoded forms of translation elongation factor eEF1A. In humans and mice eEF1A1 and eEF1A2 are 92 % identical at the amino acid level, but the well conserved developmental switch between the two variants in specific tissues suggests the existence of important functional differences. Heterozygous mutations in eEF1A2 result in neurodevelopmental disorders in humans; the mechanism of pathogenicity is unclear, but one hypothesis is that there is a dominant negative effect on eEF1A1 during development. The high degree of similarity between the eEF1A proteins has complicated expression analysis in the past; here we describe a gene edited mouse line in which we have introduced a V5 tag in the gene encoding eEF1A2. Expression analysis using anti-V5 and anti-eEF1A1 antibodies demonstrates that, in contrast to the prevailing view that eEF1A2 is only expressed postnatally, it is expressed from as early as E11.5 in the developing neural tube. Two colour immunofluorescence also reveals coordinated switching between eEF1A1 and eEF1A2 in different regions of postnatal brain. Completely reciprocal expression of the two variants is seen in post-weaning mouse brain with eEF1A1 expressed in oligodendrocytes and astrocytes and eEF1A2 in neuronal soma. Although eEF1A1 is absent from neuronal cell bodies after development, it is widely expressed in axons. This expression does not appear to coincide with myelin sheaths originating from oligodendrocytes but rather results from localised translation within the axon, suggesting that both variants are transcribed in neurons but show completely distinct subcellular localisation at the protein level. These findings will form an underlying framework for understanding how missense mutations in eEF1A2 result in neurodevelopmental disorders.

Modelling epilepsy in the mouse: challenges and solutions.

In most mouse models of disease, the outward manifestation of a disorder can be measured easily, can be assessed with a trivial test such as hind limb clasping, or can even be observed simply by comparing the gross morphological characteristics of mutant and wild-type littermates. But what if we are trying to model a disorder with a phenotype that appears only sporadically and briefly, like epileptic seizures? The purpose of this Review is to highlight the challenges of modelling epilepsy, in which the most obvious manifestation of the disorder, seizures, occurs only intermittently, possibly very rarely and often at times when the mice are not under direct observation. Over time, researchers have developed a number of ways in which to overcome these challenges, each with their own advantages and disadvantages. In this Review, we describe the genetics of epilepsy and the ways in which genetically altered mouse models have been used. We also discuss the use of induced models in which seizures are brought about by artificial stimulation to the brain of wild-type animals, and conclude with the ways these different approaches could be used to develop a wider range of anti-seizure medications that could benefit larger patient populations.

Recapitulation of the EEF1A2 D252H neurodevelopmental disorder-causing missense mutation in mice reveals a toxic gain of function.

Heterozygous de novo mutations in EEF1A2, encoding the tissue-specific translation elongation factor eEF1A2, have been shown to cause neurodevelopmental disorders including often severe epilepsy and intellectual disability. The mutational profile is unusual; ~50 different missense mutations have been identified but no obvious loss of function mutations, though large heterozygous deletions are known to be compatible with life. A key question is whether the heterozygous missense mutations operate through haploinsufficiency or a gain of function mechanism, an important prerequisite for design of therapeutic strategies. In order both to address this question and to provide a novel model for neurodevelopmental disorders resulting from mutations in EEF1A2, we created a new mouse model of the D252H mutation. This mutation causes the eEF1A2 protein to be expressed at lower levels in brain but higher in muscle in the mice. We compared both heterozygous and homozygous D252H and null mutant mice using behavioural and motor phenotyping alongside molecular modelling and analysis of binding partners. Although the proteomic analysis pointed to a loss of function for the D252H mutant protein, the D252H homozygous mice were more severely affected than null homozygotes on the same genetic background. Mice that are heterozygous for the missense mutation show no behavioural abnormalities but do have sex-specific deficits in body mass and motor function. The phenotyping of our novel mouse lines, together with analysis of molecular modelling and interacting proteins, suggest that the D252H mutation results in a gain of function.

PTSD symptoms are differentially associated with general distress and physiological arousal: Implications for the conceptualization and measurement of PTSD.

BACKGROUND: The primary purpose of this study was to examine the place of posttraumatic stress disorder (PTSD) vis-à-vis the external dimensions of general distress and physiological arousal. METHODS: Using data collected from veterans of the wars in Iraq and Afghanistan (N = 1350), latent variable covariance structure modeling was employed to compare correlations of PTSD symptom clusters and individual PTSD symptoms with general distress and physiological arousal. RESULTS: Each PTSD symptom cluster, and 17 of 20 individual PTSD symptoms were more strongly associated with general distress than with physiological arousal. However, moderate to strong associations were also found between physiological arousal and both PTSD clusters and symptoms. LIMITATIONS: Findings are based on self-reported data elicited from a single sample of veterans with substantial PTSD symptoms. Replication, particularly by clinician interview, is necessary. Generalizability to other traumatized populations is unknown. CONCLUSIONS: Results offer support, with caveats, for viewing PTSD as a distress disorder. Findings are not consistent with the position that PTSD is a hybrid disorder with some features reflecting hyperarousal and others indicative of general distress. Results have implications for the conceptualization and measurement of PTSD.

A randomized controlled trial of a web-based, personalized normative feedback alcohol intervention for young-adult veterans.

OBJECTIVE: Young-adult American veterans are at risk for problematic alcohol use. However, they are unlikely to seek care and may drop out from lengthy, multicomponent treatments when they do get care. This randomized controlled trial tested a very brief alcohol intervention delivered over the Internet to reach the population of young-adult veterans to help reduce their drinking. METHOD: Veterans (N = 784) were recruited from Facebook and randomized to either a control condition or a personalized normative feedback (PNF) intervention seeking to correct drinking perceptions of gender-specific veteran peers. RESULTS: At immediate postintervention, PNF participants reported greater reductions in their perceptions of peer drinking and intentions to drink over the next month, compared with control participants. At 1-month follow-up, PNF participants reduced their drinking behavior and related consequences to a significantly greater extent than controls. Specifically, PNF participants drank 3.4 fewer drinks per week, consumed 0.4 fewer drinks per occasion, binge drank on 1.0 fewer days, and experienced about 1.0 fewer consequences than control participants in the month after the intervention. Intervention effects for drinks per occasion were most pronounced among more problematic drinkers. Changes in perceived norms from baseline to 1-month follow-up mediated intervention efficacy. CONCLUSION: Though effects were assessed after only 1 month, findings have potential to inform broader, population-level programs designed for young veterans to prevent escalation of drinking and development of long-term alcohol problems. Given the simplicity of the PNF approach and ease of administration, this intervention has the potential for a substantial impact on public health. (PsycINFO Database Record

Recruitment and retention of young adult veteran drinkers using Facebook.

The objective of this study was to describe the feasibility of using Facebook as a platform to recruit and retain young adult veteran drinkers into an online-alcohol use intervention study. Facebook's wide accessibility and popularity among the age group that comprises the majority of veterans from the conflicts in Iraq and Afghanistan make it a compelling resource through which research can extend its reach to this otherwise hard-to-reach group. We developed a series of Facebook advertisement campaigns to reach veteran drinkers not specifically searching for alcohol treatment. In doing so, we recruited 793 valid veteran participants in approximately two weeks for an advertising cost of $4.53 per obtained participant. The study sample consisted primarily of male veterans, between 19 and 34 years of age, who were drinking at moderate to heavy levels. Although about half of the sample reported mental health comorbidity, few had received any mental health or substance use treatment in the past year. Facebook appears to be a valuable mechanism through which to recruit young veterans with unmet behavioral health needs, although more specific efforts may be needed to engage certain types of veterans after initial study enrollment.

Combat experience and problem drinking in veterans: Exploring the roles of PTSD, coping motives, and perceived stigma.

PURPOSE: The current investigation sought to illustrate the etiology of adverse alcohol consequences in young adult veterans using a path analytic framework. METHODS: A total of 312 veterans aged 19-34 were enrolled in a larger intervention study on alcohol use. At baseline, participants completed measures of combat severity, PTSD symptom severity, and drinking motives to cope. At one month follow-up, participants completed measures of perceived stigma of behavioral health treatment seeking and past 30-day alcohol consequences. RESULTS: After entering the covariates of age, gender, race/ethnicity, and past year behavioral health treatment utilization, a path analytic model demonstrated a good fit to the data predicting alcohol consequences in this population. Further, a separate exploratory analysis confirmed that both drinking motives to cope and perceived stigma of behavioral health treatment seeking mediated the link between PTSD symptom severity and alcohol consequences. CONCLUSIONS: The current model expands upon prior research showing the relationship between combat severity and alcohol use behavior in young adult veterans. Results support the notion that veterans with PTSD symptoms may drink to cope and that perceived stigma surrounding help seeking may further contribute to alcohol related problems.

Behavioral Health Treatment Receipt Among a Community Sample of Young Adult Veterans.

Young adult veterans are at risk for behavioral health problems such as depression, posttraumatic stress disorder (PTSD), and substance misuse. Despite this, studies of veterans within the Veterans Affairs Healthcare System (VA) indicate that about half of those warranting treatment receive it in any form, with few receiving an adequate dose of care. For this study, the behavioral health screening status and behavioral health usage (including care outside of VA settings) among a community sample of 812 young adult veterans recruited from the Internet is described. Although approximately 70% of the sample screened positive for behavioral health problems, only one fifth to three fifths of those screening positive reported any mental health or substance use treatment in the past year, with one third or less receiving a dose of minimally adequate psychotherapy or psychotropic care. Findings expand on prior work and suggest that more effort is necessary to engage young veterans in behavioral health services.

Study protocol for a web-based personalized normative feedback alcohol intervention for young adult veterans.

BACKGROUND: Young adult veterans from the wars in Iraq and Afghanistan represent a population at-risk for heavy and problematic alcohol use. Unfortunately, few seek treatment for alcohol concerns and those that do seek care may drop out from lengthy multicomponent treatments. Additionally, veterans who live in rural areas and those who are not engaged in the Veterans Affairs Healthcare System are often overlooked, difficult to engage in treatment, and may not be actively seeking treatment for heavy patterns of use that may develop into an alcohol use disorder. The objective of this proposed randomized controlled trial is to develop and pilot test a brief, stand-alone Internet-based alcohol intervention with young adult veterans to help them reduce their drinking and prevent the development of problematic alcohol use. METHODS/DESIGN: Recruitment and intervention is delivered entirely over the Internet to address barriers to seeking care among this at-risk group. The online intervention consists of an assessment followed by a single module of personalized normative feedback (PNF), which provides individuals with accurate information to reduce misperceptions regarding the frequency and acceptability of risky peer behavior. PNF has established efficacy as included within multicomponent interventions targeting military populations or as a stand-alone intervention with young adult college students, but has not yet been empirically supported for the at-risk veteran population. This paper describes the development of the PNF intervention content and details the protocol for the intervention study, which will utilize a sample of 600 young adult veterans to examine the efficacy of the brief PNF intervention targeted toward reducing perceived norms, intentions to drink, actual drinking behavior, and consequences. Specific subpopulations of this veteran population, including those with mental health concerns and those differentiated by level of drinking problems, reasons for drinking, and connection to peers, will be examined to support generalizability of the intervention. DISCUSSION: This intervention has the potential to improve veteran health care by utilizing a novel approach to increase access to care, assist with drinking reductions, and prevent alcohol-related problems. Trial registration ClinicalTrials.gov Identifier NCT02187887.

Diabetes and Cardiovascular Disease Risk in Cambodian Refugees.

UNLABELLED: To determine rates of diabetes, hypertension, and hyperlipidemia in Cambodian refugees, and to assess the proportion whose conditions are satisfactorily managed in comparison to the general population. Self-report and laboratory/physical health assessment data obtained from a household probability sample of U.S.-residing Cambodian refugees (N = 331) in 2010-2011 were compared to a probability sample of the adult U.S. population (N = 6,360) from the 2009-2010 National Health and Nutrition Examination Survey. Prevalence of diabetes, hypertension and hyperlipidemia in Cambodian refugees greatly exceeded rates found in the age- and gender-adjusted U.S. POPULATION: Cambodian refugees with diagnosed hypertension or hyperlipidemia were less likely than their counterparts in the general U.S. population to have blood pressure and total cholesterol within recommended levels. Increased attention should be paid to prevention and management of diabetes and cardiovascular disease risk factors in the Cambodian refugee community. Research is needed to determine whether this pattern extends to other refugee groups.

Young adult veteran perceptions of peers' drinking behavior and attitudes.

Social norms-based interventions have shown promise in reducing drinking behavior and the resulting consequences in young adults. Although most research has focused on young civilians (i.e., college students), some studies have investigated social norms-based interventions with active-duty military and veteran samples. Yet, research has not yet determined how to maximize the effectiveness of social norms-based interventions in this heavy-drinking population. As an initial step toward this goal, the current study utilized a community sample of 1,023 young adult veterans to examine (a) whether veteran perceptions of the drinking behavior of their veteran peers differ from their perceptions of civilian drinking behavior, (b) whether perceptions of specific veteran groups differ from the actual drinking behavior of veterans within those groups, (c) what levels of specificity in reference groups (same-gender civilians, same-branch veterans, same-gender veterans, or same-branch-and-gender veterans) are most strongly associated with veterans' own drinking, and (d) whether perceptions about others' attitudes toward drinking also contribute independently of perceived behavioral norms to veteran drinking. Findings indicated that participants perceived that other veterans drank more than civilians and that veteran groups drank more than veterans in the sample actually drank. Veteran-specific perceived behavioral norms were similar in their associations with drinking outcomes, whereas same-gender civilian perceived behavioral norms exhibited little or no associations with drinking. Veteran-specific perceived attitudinal norms exhibited little or no association with drinking behavior after controlling for perceived behavioral norms. These findings can be used to inform the development of social norms interventions for young adult veterans.

Help-Seeking Stigma and Mental Health Treatment Seeking Among Young Adult Veterans.

Veterans underutilize mental health services. We investigated the association between treatment seeking stigma and utilization of mental health services in a sample of 812 young adult veterans. Higher perceived public stigma of treatment seeking was significantly related to lower treatment utilization. Although many veterans were concerned about negative perceptions if they were to seek treatment, a much smaller number of them endorsed that they would judge a fellow veteran negatively in similar situation. Targeting perceived public stigma of treatment seeking, through perceived norms interventions, might help in narrowing the gap between the need and receipt of help among veterans.

Using facebook to recruit young adult veterans: online mental health research.

BACKGROUND: Veteran research has primarily been conducted with clinical samples and those already involved in health care systems, but much is to be learned about veterans in the community. Facebook is a novel yet largely unexplored avenue for recruiting veteran participants for epidemiological and clinical studies. OBJECTIVE: In this study, we utilized Facebook to recruit a sample of young adult veterans for the first phase of an online alcohol intervention study. We describe the successful Facebook recruitment process, including data collection from over 1000 veteran participants in approximately 3 weeks, procedures to verify participation eligibility, and comparison of our sample with nationally available norms. METHODS: Participants were young adult veterans aged 18-34 recruited through Facebook as part of a large study to document normative drinking behavior among a large community sample of veterans. Facebook ads were targeted toward young veterans to collect information on demographics and military characteristics, health behaviors, mental health, and health care utilization. RESULTS: We obtained a sample of 1023 verified veteran participants over a period of 24 days for the advertising price of approximately US $7.05 per verified veteran participant. Our recruitment strategy yielded a sample similar to the US population of young adult veterans in most demographic areas except for race/ethnicity and previous branch of service, which when we weighted the sample on race/ethnicity and branch a sample better matched with the population data was obtained. The Facebook sample recruited veterans who were engaged in a variety of risky health behaviors such as binge drinking and marijuana use. One fourth of veterans had never since discharge been to an appointment for physical health care and about half had attended an appointment for service compensation review. Only half had attended any appointment for a mental health concern at any clinic or hospital. Despite more than half screening positive for current probable mental health disorders such as post-traumatic stress disorder, depression, anxiety, only about 1 in 3 received mental health care in the past year and only 1 in 50 received such care within the past month. CONCLUSIONS: This work expands on the work of other studies that have examined clinical samples of veterans only and suggests Facebook can be an adequate method of obtaining samples of veterans in need of care. TRIAL REGISTRATION: Clinicaltrials.gov NCT02187887; http://clinicaltrials.gov/ct2/show/NCT02187887 (Archived by WebCite at http://www.webcitation.org/6YiUKRsXY).

Characterizing the Mental Health Care of U.S. Cambodian Refugees.

OBJECTIVE: This study examined U.S. Cambodian refugees' utilization of mental health services across provider types, levels of minimally adequate care, and mode of communication with providers. METHODS: Face-to-face household interviews about mental health service use in the past 12 months were conducted as part of a study of a probability sample of Cambodian refugees. The analytic sample was restricted to the 227 respondents who met past 12-month criteria for posttraumatic stress disorder (PTSD) or major depressive disorder or both. Analyses were weighted to account for complex sampling design effects and for attrition. RESULTS: Fifty-two percent of Cambodian refugees who met diagnostic criteria obtained mental health services in the past 12 months. Of those who obtained care, 75% visited a psychiatrist and 56% a general medical provider. Only 7% had obtained care from other mental health specialty providers. Virtually all respondents who had seen a psychiatrist (100%) or a general medical doctor (97%) had been prescribed a psychotropic medication. Forty-five percent had received minimally adequate care. Most relied on interpreters to communicate with providers. CONCLUSIONS: Cambodian refugees' rates of mental health service utilization and minimally adequate care were comparable to those of individuals in the general U.S. POPULATION: Cambodian refugees obtained care almost entirely from psychiatrists and general medical doctors, and nearly all were receiving pharmacotherapy; these findings differ from rates seen in a nationally representative sample. Given this pattern of utilization, and the persistently high levels of PTSD and depression found among Cambodian refugees, treatment improvements may require identification of creative approaches to delivering more evidence-based psychotherapy.

Access to Behavioral Health Care for Geographically Remote Service Members and Dependents in the U.S.

With many service members now returning to the United States from the recent conflicts in Iraq and Afghanistan, concern over adequate access to behavioral health care (treatment for mental, behavioral, or addictive disorders) has risen. Yet data remain very sparse regarding how many service members (and their dependents) reside in locations remote from behavioral health providers, as well as the resulting effect on their access to and utilization of care. Little is also known about the effectiveness of existing policies and other efforts to improve access to services among this population. To help fill these gaps, a team of RAND researchers conducted a geospatial analysis using TRICARE and other data, finding that roughly 300,000 military service members and 1 million dependents are geographically distant from behavioral health care, and an analysis of claims data indicated that remoteness is associated with lower use of specialty behavioral health care. A review of existing policies and programs discovered guidelines for access to care, but no systematic monitoring of adherence to those guidelines, limiting their value. RAND researchers recommend implementing a geospatial data portal and monitoring system to track access to care in the military population and mark progress toward improvements in access to care. In addition, the RAND team highlighted two promising pathways for improving access to care among remote military populations: telehealth and collaborative care that integrates primary care with specialty behavioral care.

A comparison of Cambodian-American adolescent substance use behavior to national and local norms.

INTRODUCTION: This study was designed to compare rates of alcohol, marijuana, and cigarette use in Cambodian-American adolescents with norms from nationally- and regionally-representative peers. METHODS: Substance use data from 439 10th grade Cambodian-American adolescents in Long Beach, California were compared to grade- and gender-matched nationally representative data from the Monitoring the Future study and data from the California Healthy Kids Survey of students within the same school district. RESULTS: Overall, the Cambodian-American youth were less likely than nationally- and regionally-representative youth to use alcohol, marijuana, and cigarettes. Specifically, relative to estimates obtained for the general population and students attending school in the same school district, Cambodian-American youth were significantly less likely to use alcohol and marijuana. Cambodian-American youth were also less likely than youth in the general population to smoke cigarettes, but did not differ statistically from youth within their same school district. CONCLUSIONS: As a group, Cambodian-American youth may not be at especially high risk for substance use. As is the case with virtually all populations, some individuals within the Cambodian-American group are likely to have more difficulty than others with substance use concerns. Thus, additional research is needed to identify factors that might help to identify high users with potential service needs.

Are mental disorders more common in urban than rural areas of the United States?

Urban vs. rural residence is commonly cited as a risk factor for depression and other mental disorders, but epidemiological evidence for this relationship in the US is inconclusive. We examined three consecutive annual samples (2009-2011) of adolescents (age 12-17, N = 55,583) and adults (age 18 and over, N = 116,459) from the National Survey of Drug Use and Health (NSDUH) to compare the prevalence of major depression and other serious mental illness across four categories of urbanicity: (1) large metropolitan areas, (2) small metropolitan areas, (3) semi-rural areas, and (4) rural areas, with and without adjustment for other demographic risk factors. For adolescents, no association was observed between urbanicity and the prevalence of major depression, with or without statistical adjustments. For adults, no differences were found in the prevalence of major depression or serious mental illness between large metropolitan areas and rural areas, but the prevalence of both was slightly higher in the two intermediate urbanicity categories than in large metropolitan areas, with statistically significant odds ratios after adjustment ranging from 1.12 to 1.19. Contrary to expectations, the prevalence of mental disorders was not higher in the most urban compared with the most rural areas, suggesting that the move to identify mechanistic explanations for risk associated with the urban environment is premature. Evidence of slightly higher prevalence in small urban and semi-rural areas relative to large urban areas, reported for the first time, requires additional investigation.

Prevalence of mental health problems among Iraq and Afghanistan veterans who have and have not received VA services.

OBJECTIVE: Roughly half of veterans who served in Operation Enduring Freedom and Operation Iraqi Freedom (OEF/OIF) have not received services from the Veterans Health Administration (VHA). This study assessed probable posttraumatic stress disorder (PTSD) and depression among OEF/OIF veterans by receipt of VHA services. METHODS: In 2010 a mixed-mode survey assessing symptoms and VHA services utilization was fielded in a random sample of 913 New York State OEF/OIF veterans. RESULTS: Probable PTSD and depression were roughly three times more common among veterans who had received VHA services (N=537) (PTSD, 23%; depression, 21%) than those who had not (N=376) (PTSD, 6%; depression, 8%). CONCLUSIONS: Studies of veterans receiving VHA services likely overstate the prevalence of mental health problems among the broader OEF/OIF veteran population. However, many veterans with mental health problems are not receiving VHA services. Policies that improve outreach to this population may improve health outcomes.