Genome editing in the mouse brain with minimally immunogenic Cas9 RNPs.

Transient delivery of CRISPR-Cas9 ribonucleoproteins (RNPs) into the central nervous system (CNS) for therapeutic genome editing could avoid limitations of viral vector-based delivery including cargo capacity, immunogenicity, and cost. Here, we tested the ability of cell-penetrant Cas9 RNPs to edit the mouse striatum when introduced using a convection-enhanced delivery system. These transient Cas9 RNPs showed comparable editing of neurons and reduced adaptive immune responses relative to one formulation of Cas9 delivered using AAV serotype 9. The production of ultra-low endotoxin Cas9 protein manufactured at scale further improved innate immunity. We conclude that injection-based delivery of minimally immunogenic CRISPR genome editing RNPs into the CNS provides a valuable alternative to virus-mediated genome editing.

Developing Nutrition Label Reading Skills: A Web-Based Practice Approach.

BACKGROUND: Nutrition labels offer the information needed to follow Dietary Guidelines for Americans, yet many individuals use labels infrequently or ineffectively due to limited comprehension and the effort required to use them. OBJECTIVE: The objective of our study was to develop and test a Web-based label-reading training tool to improve individuals' ability to use labels to select more healthful foods. We were particularly interested in determining whether practice can lead to increased accuracy using labels as well as decreased effort, together reflecting greater efficiency. We compared a basic and an enhanced, prior-knowledge version of the tool that contained an additional component, a brief nutrition tutorial. METHODS: Participants were 140 college students with an average age of 20.7 (SD 2.1) years and education 14.6 (SD 1.2) years, who completed 3 sets of practice that were designed to teach them, through repetition and feedback, how to use nutrition labels to select more healthful products. Prior to training, participants in the prior-knowledge group viewed a multimedia nutrition presentation, which those in the basic group did not receive. Mixed-effects models tested for improvement in accuracy and speed with practice, and whether improvements varied by group. RESULTS: The training led to significant increases in average accuracy across the 3 practice sets (averaging 79% [19/24 questions], 92% [22/24], 96% [23/24] respectively, P<.001), as well as decreases in time to complete with mean (SD) values of 8.7 (2.8), 4.6 (1.8), and 4.1 (1.7) seconds, respectively. In block 3, the odds of a correct answer for the prior-knowledge group were 79% higher (odds ratio, OR=1.79, 95% CI 1.1-2.9) than those for the basic group (P=.02). There was no significant difference between the groups in block 2 (P=.89). CONCLUSIONS: Practice led to improvements in nutrition label reading skills that are indicative of early stages of automatic processing. To the extent that automatic processes are at the core of healthy habit change, this may be an efficient way to improve dietary decision-making.

Age differences in the use of serving size information on food labels: numeracy or attention?

OBJECTIVE: The ability to use serving size information on food labels is important for managing age-related chronic conditions such as diabetes, obesity and cancer. Past research suggests that older adults are at risk for failing to accurately use this portion of the food label due to numeracy skills. However, the extent to which older adults pay attention to serving size information on packages is unclear. We compared the effects of numeracy and attention on age differences in accurate use of serving size information while individuals evaluated product healthfulness. DESIGN: Accuracy and attention were assessed across two tasks in which participants compared nutrition labels of two products to determine which was more healthful if they were to consume the entire package. Participants' eye movements were monitored as a measure of attention while they compared two products presented side-by-side on a computer screen. Numeracy as well as food label habits and nutrition knowledge were assessed using questionnaires. SETTING: Sacramento area, California, USA, 2013-2014. SUBJECTS: Stratified sample of 358 adults, aged 20-78 years. RESULTS: Accuracy declined with age among those older adults who paid less attention to serving size information. Although numeracy, nutrition knowledge and self-reported food label use supported accuracy, these factors did not influence age differences in accuracy. CONCLUSIONS: The data suggest that older adults are less accurate than younger adults in their use of serving size information. Age differences appear to be more related to lack of attention to serving size information than to numeracy skills.

Strategies to uncover undiagnosed HIV infection among heterosexuals at high risk and link them to HIV care with high retention: a "seek, test, treat, and retain" study.

BACKGROUND: Over 50,000 individuals become infected with HIV annually in the U.S., and over a quarter of HIV infected individuals are heterosexuals. Undiagnosed HIV infection, as well as a lack of retention in care among those diagnosed, are both primary factors contributing to ongoing HIV incidence. Further, there are racial/ethnic disparities in undiagnosed HIV and engagement in care, with African Americans/Blacks and Latinos remaining undiagnosed longer and less engaged in care than Whites, signaling the need for culturally targeted intervention approaches to seek and test those with undiagnosed HIV infection, and link them to care with high retention. METHODS/DESIGN: The study has two components: one to seek out and test heterosexuals at high risk for HIV infection, and another to link those found infected to HIV care with high retention. We will recruit sexually active African American/Black and Latino adults who have opposite sex partners, negative or unknown HIV status, and reside in locations with high poverty and HIV prevalence. The "Seek and Test" component will compare the efficacy and cost effectiveness of two strategies to uncover undiagnosed HIV infection: venue-based sampling and respondent-driven sampling (RDS). Among those recruited by RDS and found to have HIV infection, a "Treat and Retain" component will assess the efficacy of a peer-driven intervention compared to a control arm with respect to time to an HIV care appointment and health indicators using a cluster randomized controlled trial design to minimize contamination. RDS initial seeds will be randomly assigned to the intervention or control arm at a 1:1 ratio and all recruits will be assigned to the same arm as the recruiter. Participants will be followed for 12 months with outcomes assessed using medical records and biomarkers, such as HIV viral load. DISCUSSION: Heterosexuals do not test for HIV as frequently as and are diagnosed later than other risk groups. The study has the potential to contribute an efficient, innovative, and sustainable multi-level recruitment approach and intervention to the HIV prevention portfolio. Because the majority of heterosexuals at high risk are African American/Black or Latino, the study has great potential to reduce racial/ethnic disparities in HIV/AIDS. TRIAL REGISTRATION: ClinicalTrials.gov, NCT01607541, Registered May 23, 2012.

Behavioral intervention improves treatment outcomes among HIV-infected individuals who have delayed, declined, or discontinued antiretroviral therapy: a randomized controlled trial of a novel intervention.

Nationally up to 60 % of persons living with HIV are neither taking antiretroviral therapy (ART) nor well engaged in HIV care, mainly racial/ethnic minorities. This study examined a new culturally targeted multi-component intervention to address emotional, attitudinal, and social/structural barriers to ART initiation and HIV care. Participants (N = 95) were African American/Black and Latino adults with CD4 < 500 cells/mm(3) not taking ART, randomized 1:1 to intervention or control arms, the latter receiving treatment as usual. Primary endpoints were adherence, evaluated via ART concentrations in hair samples, and HIV viral load suppression. The intervention was feasible and acceptable. Eight months post-baseline, intervention participants tended to be more likely to evidence "good" (that is, 7 days/week) adherence (60 vs. 26.7 %; p = 0.087; OR = 3.95), and had lower viral load levels than controls (t(22) = 2.29, p = 0.032; OR = 5.20), both large effect sizes. This highly promising intervention merits further study.

Misunderstanding of Front-Of-Package Nutrition Information on US Food Products.

Front-of-package nutrition symbols (FOPs) are presumably readily noticeable and require minimal prior nutrition knowledge to use. Although there is evidence to support this notion, few studies have focused on Facts Up Front type symbols which are used in the US. Participants with varying levels of prior knowledge were asked to view two products and decide which was more healthful. FOPs on packages were manipulated so that one product was more healthful, allowing us to assess accuracy. Attention to nutrition information was assessed via eye tracking to determine what if any FOP information was used to make their decisions. Results showed that accuracy was below chance on half of the comparisons despite consulting FOPs. Negative correlations between attention to calories, fat, and sodium and accuracy indicated that consumers over-relied on these nutrients. Although relatively little attention was allocated to fiber and sugar, associations between attention and accuracy were positive. Attention to vitamin D showed no association to accuracy, indicating confusion surrounding what constitutes a meaningful change across products. Greater nutrition knowledge was associated with greater accuracy, even when less attention was paid. Individuals, particularly those with less knowledge, are misled by calorie, sodium, and fat information on FOPs.

Relationships among food label use, motivation, and dietary quality.

Nutrition information on packaged foods supplies information that aids consumers in meeting the recommendations put forth in the US Dietary Guidelines for Americans such as reducing intake of solid fats and added sugars. It is important to understand how food label use is related to dietary intake. However, prior work is based only on self-reported use of food labels, making it unclear if subjective assessments are biased toward motivational influences. We assessed food label use using both self-reported and objective measures, the stage of change, and dietary quality in a sample of 392 stratified by income. Self-reported food label use was assessed using a questionnaire. Objective use was assessed using a mock shopping task in which participants viewed food labels and decided which foods to purchase. Eye movements were monitored to assess attention to nutrition information on the food labels. Individuals paid attention to nutrition information when selecting foods to buy. Self-reported and objective measures of label use showed some overlap with each other (r=0.29, p<0.001), and both predicted dietary quality (p<0.001 for both). The stage of change diminished the predictive power of subjective (p<0.09), but not objective (p<0.01), food label use. These data show both self-reported and objective measures of food label use are positively associated with dietary quality. However, self-reported measures appear to capture a greater motivational component of food label use than do more objective measures.

HIV-Infected Individuals Who Delay, Decline, or Discontinue Antiretroviral Therapy: Comparing Clinic- and Peer-Recruited Cohorts.

A substantial proportion of persons living with HIV/AIDS (PLHA) delay, decline, or discontinue antiretroviral therapy (ART) when it is medically indicated (40-45%), largely African-Americans and Latinos/Hispanics. This study explores the feasibility of locating PLHA, who are not on ART (PLHA-NOA) through clinics and peer-referral; compares the two cohorts on multi-level barriers to ART; and examines readiness to initiate/reinitiate ART, a predictor of treatment outcomes. We recruited adult HIV-infected African-American and Latino/Hispanic PLHA-NOA through HIV hospital clinics and peer-referral in 2012-2013. Participants were engaged in structured 1-h assessments with reliable/valid measures on barriers to ART. We found that recruitment through peers (63.2%, 60/95) was more feasible than in clinics (36.8%, 35/90). Participants were 48.0 years old and had lived with HIV for 14.7 years on average, and 56.8% had taken ART previously. Most (61.1%) were male and African-American (76.8%), and 23.2% were Latino/Hispanic. Peer-recruited participants were older, had lived with HIV longer, were less engaged in HIV care, and were more likely to have taken ART previously. The cohorts differed in reasons for discontinuing ART. Levels of ART knowledge were comparable between cohorts (68.5% correct), and there were no differences in attitudes toward ART (e.g., mistrust), which were in the neutral range. In bivariate linear regression, readiness for ART was negatively associated with physician mistrust (B = -10.4) and positively associated with self-efficacy (B = 5.5), positive outcome expectancies (B = 6.3), beliefs about personal necessity of ART (B = 17.5), and positive internal norms (B = 7.9). This study demonstrates the feasibility of engaging this vulnerable population through peer-referral. Peer-recruited PLHA evidence particularly high rates of risk factors compared to those in hospital clinics. Interventions to support ART initiation and continuation are sorely needed for both subgroups.

Natural versus commercial carbohydrate supplementation and endurance running performance.

BACKGROUND: We examined the metabolic, performance and gastrointestinal (GI) effects of supplementation with a natural food product (raisins) compared to a commercial product (sport chews). METHODS: Eleven male (29.3 ± 7.9 yrs; mean and SD) runners completed three randomized trials (raisins, chews and water only) separated by seven days. Each trial consisted of 80-min (75%VO2max) treadmill running followed by a 5-km time trial (TT). Heart rate (HR), respiratory exchange ratio (RER), blood lactate, serum free fatty acids (FFA), glycerol and insulin, plasma glucose and creatine kinase, GI symptoms and rating of perceived exertion (RPE) were recorded every 20-min. We employed a within-subject two-way analysis of variance (ANOVA) for repeated measures with a Fisher's post hoc analysis to determine significant differences. RESULTS: VO2, HR, lactate, glycerol and RPE did not differ due to treatment. Average plasma glucose was maintained at resting levels (5.3 ± 0.4 mmol·L-1) during the sub-maximal exercise bout (5.9 ± 0.6, 5.7 ± 0.6 and 5.5 ± 0.5 mmol·L-1 for chews, raisins and water respectively), and was significantly higher with chews than water only. RER and % of non-protein macronutrient oxidation derived from carbohydrate was highest with chews, followed by raisins and water was the lowest (74.4 ± 6.4, 70.0 ± 7.0 and 65.1 ± 8.7% for chews, raisins and water respectively) during the sub-maximal exercise period. Serum FFA was higher in the water treatment versus both raisins and chews at 80 min of sub-maximal exercise. Serum insulin was higher with the chews than both raisins and water (5.1 ± 2.0, 3.1 ± 0.8, 1.9 ± 0.6 uU·ml-1 for chews, raisins and water respectively). Plasma creatine kinase, corrected for baseline values, for the last 40 min of the sub-maximal exercise bout, was higher with raisins compared to other treatments. The TT was faster for both carbohydrate supplements (20.6 ± 2.6, 20.7 ± 2.5, 21.6 ± 2.7 min for raisin, chews and water respectively). GI disturbance was mild for all treatments. CONCLUSION: Raisins and chews promoted higher carbohydrate oxidation and improved running performance compared to water only. Running performance was similar between the raisins and chews, with no significant GI differences.

Mechanisms underlying comprehension of health information in adulthood: the roles of prior knowledge and working memory capacity.

Prior knowledge, working memory capacity (WMC), and conceptual integration (attention allocated to integrating concepts in text) are critical within many contexts; however, their impact on the acquisition of health information (i.e. learning) is relatively unexplored.We examined how these factors impact learning about nutrition within a cross-sectional study of adults ages 18 to 81. Results showed that conceptual integration mediated the effects of knowledge and WMC on learning, confirming that attention to concepts while reading is important for learning about health. We also found that when knowledge was controlled, age declines in learning increased, suggesting that knowledge mitigates the effects of age on learning about nutrition.

Predictors of nutrition information comprehension in adulthood.

OBJECTIVE: The goal of the present study was to examine relationships among several predictors of nutrition comprehension. We were particularly interested in exploring whether nutrition knowledge or motivation moderated the effects of attention on comprehension across a wide age range of adults. METHODS: Ninety-three participants, ages 18-80, completed measures of nutrition knowledge and motivation and then read nutrition information (from which attention allocation was derived) and answered comprehension questions. RESULTS: In general, predictor variables were highly intercorrelated. However, knowledge, but not motivation, had direct effects on comprehension accuracy. In contrast, motivation influenced attention, which in turn influenced accuracy. Results also showed that comprehension accuracy decreased-and knowledge increased-with age. When knowledge was statistically controlled, age declines in comprehension increased. CONCLUSION: Knowledge is an important predictor of nutrition information comprehension and its role increases in later life. Motivation is also important; however, its effects on comprehension differ from knowledge. PRACTICE IMPLICATIONS: Health educators and clinicians should consider cognitive skills such as knowledge as well as motivation and age of patients when deciding how to best convey health information. The increased role of knowledge among older adults suggests that lifelong educational efforts may have important payoffs in later life.

Effects of an amino acid carbohydrate drink on exercise performance after consecutive-day exercise bouts.

This study examined the effect of amino acids in a carbohydrate beverage on cycling performance. Twelve male athletes (28.5 +/- 2.1 yr) cycled at 75% VO2peak for 90 min followed by a ride to exhaustion at 85% VO2peak, before (T1) and on 2 consecutive days (T2 and T3) after 2 weeks of supplementation with 3.6% carbohydrate plus 1% amino acids (AA) or 4.6% carbohydrate-only (CHO) isocaloric beverages. Muscle damage was assessed by plasma creatine kinase (CK), and muscle fatigue by changes in vertical jump pre- to postexercise. Muscle soreness, overall fatigue, and changes in mood state were assessed using questionnaires. Plasma CK was lower for AA in T3 (214.0 +/- 13.5 vs. 485.9 +/- 191.4 U/L immediately post, 213.9 +/- 13.1 vs. 492.0 +/- 199.4 U/L 5 hr post, and 194.9 +/- 17.9 vs. 405.9 +/- 166.6 U/L 24 hr postexercise in AA and CHO, respectively). Time to exhaustion decreased from T2 to T3 only in CHO (10.9 +/- 2.5 to 12.6 +/- 3.2 vs. 13.8 +/- 2.8 to 7.8 +/- 1.5 min in AA and CHO, respectively). Vertical-jump change from pre- to postexercise was greater in T3 for the CHO treatment. Total fatigue score and mood disturbance decreased significantly only with AA in T3. The addition of AA to a carbohydrate beverage after consecutive-day exercise bouts reduced muscle damage as indicated by CK levels, decreased fatigue, and maintained exercise performance compared with consuming carbohydrate alone.

Carbohydrate-supplement form and exercise performance.

Numerous studies have shown that ingesting carbohydrate in the form of a drink can improve exercise performance by maintaining blood glucose levels and sparing endogenous glycogen stores. The effectiveness of carbohydrate gels or jellybeans in improving endurance performance has not been examined. On 4 separate days and 1-2 hr after a standardized meal, 16 male (8; 35.8 +/- 2.5 yr) and female (8; 32.4 +/- 2.4 yr) athletes cycled at 75% VO(2peak) for 80 min followed by a 10-km time trial. Participants consumed isocaloric (0.6 g of carbohydrate per kg per hour) amounts of randomly assigned sports beans, sports drink, gel, or water only, before, during, and after exercise. Blood glucose concentrations were similar at rest between treatments and decreased significantly during exercise with the water trial only. Blood glucose concentrations for all carbohydrate supplements were significantly, p < .05, higher than water during the 80-min exercise bout and during the time trial (5.7 +/- 0.2 mmol/L for sports beans, 5.6 +/- 0.2 mmol/L for sports drink, 5.7 +/- 0.3 mmol/L for gel, and 4.6 +/- 0.3 mmol/L for water). There were no significant differences in blood glucose between carbohydrate treatments. The 10-km time trials using all 3 carbohydrate treatments were significantly faster (17.2 +/- 0.6 min for sports beans, 17.3 +/- 0.6 min for sports drink, and 17.3 +/- 0.6 min for gel) than water (17.8 +/- 0.7 min). All carbohydrate-supplement types were equally effective in maintaining blood glucose levels during exercise and improving exercise performance compared with water only.