Virally programmed extracellular vesicles sensitize cancer cells to oncolytic virus and small molecule therapy.

Recent advances in cancer therapeutics clearly demonstrate the need for innovative multiplex therapies that attack the tumour on multiple fronts. Oncolytic or "cancer-killing" viruses (OVs) represent up-and-coming multi-mechanistic immunotherapeutic drugs for the treatment of cancer. In this study, we perform an in-vitro screen based on virus-encoded artificial microRNAs (amiRNAs) and find that a unique amiRNA, herein termed amiR-4, confers a replicative advantage to the VSVΔ51 OV platform. Target validation of amiR-4 reveals ARID1A, a protein involved in chromatin remodelling, as an important player in resistance to OV replication. Virus-directed targeting of ARID1A coupled with small-molecule inhibition of the methyltransferase EZH2 leads to the synthetic lethal killing of both infected and uninfected tumour cells. The bystander killing of uninfected cells is mediated by intercellular transfer of extracellular vesicles carrying amiR-4 cargo. Altogether, our findings establish that OVs can serve as replicating vehicles for amiRNA therapeutics with the potential for combination with small molecule and immune checkpoint inhibitor therapy.

Gestational Folate and Folic Acid Intake among Women in Canada at Higher Risk of Pre-Eclampsia.

BACKGROUND: Periconceptional folic acid (FA) supplementation is recommended to prevent neural tube defects; however, the extent to which recommendations are met through dietary sources and supplements is not clear. OBJECTIVES: Our objective was to evaluate the dietary and supplemental intakes of FA in a Canadian pregnancy cohort and to determine the proportions of pregnant women exceeding the Estimated Average Requirement (EAR) and Tolerable Upper Intake Level (UL). METHODS: FACT (the Folic Acid Clinical Trial) was an international multicenter, randomized, double-blinded, placebo-controlled, phase III trial investigating FA for the prevention of pre-eclampsia in high-risk pregnancies. Participants were enrolled from Canadian sites at 8-16 weeks of gestation. Dietary and supplemental FA intake data were collected through participant interviews and FFQs at the time of FACT enrollment. Categorical data were summarized as n (%) and continuous data as median (IQR). RESULTS: This study included 1198 participants. Participants consumed 485 µg dietary folate equivalents (DFE)/d (IQR: 370-630 µg DFE/d) from dietary sources of folate and FA. Through diet alone, 43.4% of participants consumed ≥520 µg DFE/d, the EAR for pregnant individuals. Of the 91.9% of participants who consumed daily FA supplements, 0.4% consumed <400 µg FA/d and 96.0% consumed ≥1000 µg/d, the UL for FA. Median (IQR) total folate intake was 2167 µg DFE/d (2032-2325 µg DFE/d); 95.3% of participants met or exceeded the EAR from all sources, but 1069 (89.2%) participants exceeded the UL. CONCLUSIONS: The majority of participants in this Canadian pregnancy cohort did not consume the recommended amount of folate from dietary sources. However, most prenatal supplements contained 1000 µg FA, resulting in the majority of women exceeding the UL. With no additional benefit associated with FA intakes beyond the UL for most women, modification of prenatal supplement formulations may be warranted to ensure women meet but do not exceed recommended FA intakes.FACT was registered at clinicaltrials.gov as NCT01355159 and at isrctn.com as ISRCTN23781770.

Absolute measurement of the tissue origins of cell-free DNA in the healthy state and following paracetamol overdose.

BACKGROUND: Despite the emergence of cell-free DNA (cfDNA) as a clinical biomarker in cancer, the tissue origins of cfDNA in healthy individuals have to date been inferred only by indirect and relative measurement methods, such as tissue-specific methylation and nucleosomal profiling. METHODS: We performed the first direct, absolute measurement of the tissue origins of cfDNA, using tissue-specific knockout mouse strains, in both healthy mice and following paracetamol (APAP) overdose. We then investigated the utility of total cfDNA and the percentage of liver-specific cfDNA as clinical biomarkers in patients presenting with APAP overdose. RESULTS: Analysis of cfDNA from healthy tissue-specific knockout mice showed that cfDNA originates predominantly from white and red blood cell lineages, with minor contribution from hepatocytes, and no detectable contribution from skeletal and cardiac muscle. Following APAP overdose in mice, total plasma cfDNA and the percentage fraction originating from hepatocytes increased by ~ 100 and ~ 19-fold respectively. Total cfDNA increased by an average of more than 236-fold in clinical samples from APAP overdose patients with biochemical evidence of liver injury, and 18-fold in patients without biochemically apparent liver injury. Measurement of liver-specific cfDNA, using droplet digital PCR and methylation analysis, revealed that the contribution of liver to cfDNA was increased by an average of 175-fold in APAP overdose patients with biochemically apparent liver injury compared to healthy subjects, but was not increased in overdose patients with normal liver function tests. CONCLUSIONS: We present a novel method for measurement of the tissue origins of cfDNA in healthy and disease states and demonstrate the potential of cfDNA as a clinical biomarker in APAP overdose.

Genetic epidemiology of motor neuron disease-associated variants in the Scottish population.

Genetic understanding of motor neuron disease (MND) has evolved greatly in the past 10 years, including the recent identification of association between MND and variants in TBK1 and NEK1. Our aim was to determine the frequency of pathogenic variants in known MND genes and to assess whether variants in TBK1 and NEK1 contribute to the burden of MND in the Scottish population. SOD1, TARDBP, OPTN, TBK1, and NEK1 were sequenced in 441 cases and 400 controls. In addition to 44 cases known to carry a C9orf72 hexanucleotide repeat expansion, we identified 31 cases and 2 controls that carried a loss-of-function or pathogenic variant. Loss-of-function variants were found in TBK1 in 3 cases and no controls and, separately, in NEK1 in 3 cases and no controls. This study provides an accurate description of the genetic epidemiology of MND in Scotland and provides support for the contribution of both TBK1 and NEK1 to MND susceptibility in the Scottish population.

Physiological and perceptual responses to affect-regulated exercise in healthy young women.

The purpose of the study was to objectively measure the exercise intensity associated with affective responses of "good" and "fairly good." In Study 1, 8 active females completed 20 min of affect-regulated exercise to feel "good" or "fairly good" (order counterbalanced) followed by an intensity replication session. On-line gas analysis was used during the replication session to measure the physiological cost of exercising. In Study 2, 10 females completed either 3 trials of exercise to feel "good" (n = 5) or 3 trials to feel "fairly good" (n = 5). Each trial consisted of an affect-regulated session followed by a replication session. Across studies, the intensity to feel "fairly good" was significantly higher than to feel "good." Both intensities lay close to ventilatory threshold. The results add to evidence that women can use affect to regulate intensity and exercise at an intensity that would confer fitness and health benefits if maintained.

Motivational interviewing for screening and feedback and encouraging lifestyle changes to reduce relative weight in 4-8 year old children: design of the MInT study.

BACKGROUND: Because parental recognition of overweight in young children is poor, we need to determine how best to inform parents that their child is overweight in a way that enhances their acceptance and supports motivation for positive change. This study will assess 1) whether weight feedback delivered using motivational interviewing increases parental acceptance of their child's weight status and enhances motivation for behaviour change, and 2) whether a family-based individualised lifestyle intervention, delivered primarily by a MInT mentor with limited support from "expert" consultants in psychology, nutrition and physical activity, can improve weight outcomes after 12 and 24 months in young overweight children, compared with usual care. METHODS/DESIGN: 1500 children aged 4-8 years will be screened for overweight (height, weight, waist, blood pressure, body composition). Parents will complete questionnaires on feeding practices, physical activity, diet, parenting, motivation for healthy lifestyles, and demographics. Parents of children classified as overweight (BMI > or = CDC 85th) will receive feedback about the results using Motivational interviewing or Usual care. Parental responses to feedback will be assessed two weeks later and participants will be invited into the intervention. Additional baseline measurements (accelerometry, diet, quality of life, child behaviour) will be collected and families will be randomised to Tailored package or Usual care. Parents in the Usual care condition will meet once with an advisor who will offer general advice regarding healthy eating and activity. Parents in the Tailored package condition will attend a single session with an "expert team" (MInT mentor, dietitian, physical activity advisor, clinical psychologist) to identify current challenges for the family, develop tailored goals for change, and plan behavioural strategies that best suit each family. The mentor will continue to provide support to the family via telephone and in-person consultations, decreasing in frequency over the two-year intervention. Outcome measures will be obtained at baseline, 12 and 24 months. DISCUSSION: This trial offers a unique opportunity to identify effective ways of providing feedback to parents about their child's weight status and to assess the efficacy of a supportive, individualised early intervention to improve weight outcomes in young children. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry ACTRN12609000749202.

Pleasant for some and unpleasant for others: a protocol analysis of the cognitive factors that influence affective responses to exercise.

BACKGROUND: At exercise intensities around ventilatory threshold (VT), the extent to which individuals experience pleasure or displeasure from the exercise varies between individuals. One source of this variability is proposed to be the cognitive appraisal that occurs during the exercise which influences the generation of the affective response. When individuals self-select their own intensity they choose to exercise around VT and experience more positive affective responses, again the explanation being that cognitive appraisal processes influence the choice of intensity and resulting affective response. However, the specific factors that comprise this appraisal process have not been thoroughly explored. In addition, it is not clear if activity status influences this appraisal and different cognitive factors play a role in the generation of affective responses. Therefore, the purpose of this study was to explore the cognitive factors that influence the affective response experienced during prescribed and self-selected intensity exercise in low-active and high-active women. METHODS: Seventeen low-active and 15 high-active women (M age = 45 years, SD = 10) completed a graded exercise test and two 30 min bouts of treadmill exercise, one at a self-selected intensity and one prescribed at an intensity around VT. Using 'think aloud' procedures, every five min, the women were asked to provide an affective response and explain the thought processes that caused them to report that affective response. Using inductive content analysis, the verbal reports provided by the women were analysed for key themes and categories that emerged as explaining the factors that underpinned the generation of the affective response. Data from the low-active and high-active women were analysed separately. RESULTS: Concepts relating to pre-exercise affective state, perceptions of ability, immediate and anticipated outcomes, attentional focus and perceptions of control emerged. The physiological demands of the exercise stimulus influenced affective responses through themes related to the interpretation of physiological symptoms and the physiological state of the body. No thematic differences emerged between high-active and low-active women. CONCLUSION: Results highlight the complex interaction of psychological and physiological influences in producing an affective response to exercise and provide insight into how exercise can be structured to elicit positive affective responses.

Athlete burnout in elite sport: a self-determination perspective.

Using self-determination theory (Deci & Ryan, 1985) as the theoretical framework, we examined potential antecedents of athlete burnout in 201 elite Canadian athletes (121 females, 80 males; mean age 22.9 years). Employing a cross-sectional design, our primary aims were to investigate the relationships between behavioural regulations and athlete burnout and to examine whether self-determined motivation mediated relationships between basic needs satisfaction and athlete burnout. Our self-determination theory-derived hypotheses were largely supported. Relationships among athlete burnout and behavioural regulations mostly varied according to their rank on the self-determination continuum, with less self-determined motives showing positive associations and more self-determined motives showing negative correlations with burnout. The basic needs of competence and autonomy, plus self-determined motivation, accounted for significant amounts of variance in athlete burnout symptoms (exhaustion, R(2) = 0.31; devaluation, R(2) = 0.49; reduced accomplishment, R(2) = 0.61; global burnout, R(2) = 0.74). Self-determined motivation fully mediated the relationships that competence and autonomy had with exhaustion. Analyses showed indirect relationships between these two needs and devaluation, through their associations with self-determined motivation. Motivation partially mediated the needs-reduced sense of accomplishment relationships, but the direct effects were more prominent than the indirect effects.

Can the feeling scale be used to regulate exercise intensity?

INTRODUCTION/PURPOSE: It is important that individuals experience a positive affective response during exercise to encourage future behavior. Exercise intensity is a determinant of the affective response. Current research protocols have failed to find the intensity at which all individuals will experience this positive affective response. Consequently, the purpose of this study was to evaluate whether sedentary women could self-regulate their exercise intensity during exercise using the Hardy and Rejeski Feeling Scale (FS) to experience a specific positive affective state and to examine the specific intensities chosen and their consistency over exercise bouts. METHODS: Seventeen sedentary women completed eight 30-min laboratory-based treadmill exercise sessions (two sessions were completed each week). In four consecutive sessions, participants exercised at an intensity they perceived corresponded to an FS value of 1 (fairly good) and the other four sessions at an intensity corresponding to an FS value of 3 (good). Measures of exercise intensity were recorded. RESULTS: Participants exercised at a lower intensity to achieve an affective state of good (FS 3) compared with fairly good (FS 1). Both these intensities lay close to the individual's ventilatory threshold. The selected intensity was consistent across trials with intensity increasing across time to maintain the required affective state. CONCLUSIONS: Sedentary women can regulate intensity using the FS to experience a pleasant affective state, and the intensities chosen are physiologically beneficial for health and fitness.

The behavioral regulation in sport questionnaire (BRSQ): instrument development and initial validity evidence.

The purpose of the four studies described in this article was to develop and test a new measure of competitive sport participants' intrinsic motivation, extrinsic motivation, and amotivation (self-determination theory; Deci & Ryan, 1985). The items for the new measure, named the Behavioral Regulation in Sport Questionnaire (BRSQ), were constructed using interviews, expert review, and pilot testing. Analyses supported the internal consistency, test-retest reliability, and factorial validity of the BRSQ scores. Nomological validity evidence was also supportive, as BRSQ subscale scores were correlated in the expected pattern with scores derived from measures of motivational consequences. When directly compared with scores derived from the Sport Motivation Scale (SMS; Pelletier, Fortier, Vallerand, Tuson, & Blais, 1995) and a revised version of that questionnaire (SMS-6; Mallett, Kawabata, Newcombe, Otero-Forero, & Jackson, 2007), BRSQ scores demonstrated equal or superior reliability and factorial validity as well as better nomological validity.

A sensitive and quantitative assay for normal PrP in plasma.

Transmissible spongiform encephalopathies can be transmitted by blood transfusion. The risk of spreading the disease among the human population could be mitigated with the implementation of a blood screening assay. We developed a two-antibody assay for PrP detection in plasma using the ORIGEN technology with a protocol modification to improve the limit of detection and to increase the sample volume assayed. In the standard 200 microL format, the assay had a detection limit of 7-10 pg of recombinant PrP and 3 pg in 1 mL final volume implementation. PrP concentration measured in normal and scrapie-infected hamster brains was 7.5+/-0.9 and 57.3+/-9.6 microg/g, respectively. After a concentration step with an immuno-affinity resin, plasma PrP(c) was detected by Western blot and its concentration was measured at 3.5+/-0.8 ng/mL. From these data and assuming that blood has the same specific infectivity as brain, we estimated the concentration of abnormal PrP in hamster-infected plasma to be 32 f g/mL. The assay also detected abnormal brain PrP spiked into plasma although the limit of detection was affected. This is a novel and sensitive assay for the detection of PrP in plasma that could be developed into a platform for a plasma-based TSE test.

Mood, illness and injury responses and recovery with adventure racing.

OBJECTIVE: Exercise stress, immune status, and mood are interrelated. The stress of adventure racing is unique; exercise is very prolonged and competitive, with severe sleep deprivation and sustained cognitive demands, usually in arduous terrain and environmental conditions. The purpose of this prospective, descriptive study was to identify mood changes along with symptoms of illness and injury during and in the weeks following an international-level adventure race. METHODS: Mood, sleep, injury, and illness data were collected using questionnaires before, during, and for 2 weeks following New Zealand's Southern Traverse Adventure Race in November 2003. RESULTS: Mood was variable between athletes, but peaks of altered mood subscores were evident (P < .05) during the first 24 hours of racing, around race completion, and, as was hypothesized, 3 days after racing. Altered mood subscores resolved within 2 weeks. Symptoms of upper respiratory illness were most common immediately before (25/60, 42%) and after (28/49, 57%) racing, and largely resolved over the 2-week follow-up (5/27, 19%). Skin wounds and infections were common (43/49, 88%) immediately after the race but settled quickly. Pain was universal (100%), and musculoskeletal injury was common (38/48, 79%). Gastrointestinal complaints were common at the finish (8/49, 16%) and during the next 5 days but settled more quickly than upper respiratory symptoms. CONCLUSIONS: Adventure racing of approximately 100 hours causes significant symptomatic injury and illness and mood state disruption, which generally resolve within a fortnight following racing. Disrupted mood and symptoms of illness and injury indicate athlete susceptibility to overreaching or overtraining without sufficient recovery.

Dentistas são qualificados e motivados para cuidar de DTM em crianças e adolescentes? / Are dental surgeons qualified and motivated to care for TDJ in children and adolescents?

Este trabalho objetiva caracterizar, em uma população de cirurgiões-dentistas que atendem crianças e adolescentes, a formação para o cuidado das DTMs e o interesse despertado na graduação para o tema. Em um congresso de Odontopediatria, de 181 profissionais, 20% faziam diagnóstico de DTM raramente ou nunca, 62% ocasionalmente e 16% com freqüência. Embora o tema "DTM" desperte muito interesse para 44,19% dos sujeitos, somente 4,97% correlacionaram a motivação com a graduação, na qual a DTM foi abordada superficialmente (70,16%) ou nunca (15,46%). Quanto à DTM em crianças e adolescentes, o tema nunca foi discutido (54%) ou o foi superficialmente (38%). É relevante capacitar os cirurgiões-dentistas, na graduação, para o cuidado da DTM em todas as idades e, em especial, na infância, considerando o grande potencial preventivo de lesões mais graves.

This study aims at characterizing, in a population of dental surgeons who attend children and adolescents, the preparing for caring for TMD, as well as the interest raised for the subject during pre-graduation. In a Congress of Odontopaediatrics, of 181 professionals, 20% diagnosed TMD rarely or never, 62% occasionaly and 16% frequently. Although 44,19% were highly interested in TMD, only 4,97% had been motivacted by the subject in pre-graduation, where TMD was inserted superficially (70,16%) and never (15,46%). TMD in children and adolescents was never discussed (54%) or was superficially (38%). It is relevant to prepair dentists during pre-graduation for caring for this affection in patients of all ages and, specially, in children, considering the great potential for prevention of more severe lesions.