Mouse Model of Fragile X Syndrome Analyzed by Quantitative Proteomics: A Comparison of Methods.

Multiple methods for quantitative proteomics are available for proteome profiling. It is unclear which methods are most useful in situations involving deep proteome profiling and the detection of subtle distortions in the proteome. Here, we compared the performance of seven different strategies in the analysis of a mouse model of Fragile X Syndrome, involving the knockout of the fmr1 gene that is the leading cause of autism spectrum disorder. Focusing on the cerebellum, we show that data-independent acquisition (DIA) and the tandem mass tag (TMT)-based real-time search method (RTS) generated the most informative profiles, generating 334 and 329 significantly altered proteins, respectively, although the latter still suffered from ratio compression. Label-free methods such as BoxCar and a conventional data-dependent acquisition were too noisy to generate a reliable profile, while TMT methods that do not invoke RTS showed a suppressed dynamic range. The TMT method using the TMTpro reagents together with complementary ion quantification (ProC) overcomes ratio compression, but current limitations in ion detection reduce sensitivity. Overall, both DIA and RTS uncovered known regulators of the syndrome and detected alterations in calcium signaling pathways that are consistent with calcium deregulation recently observed in imaging studies. Data are available via ProteomeXchange with the identifier PXD039885.

High-Efficiency Enrichment by Saturating Nanoliters of Protein Affinity Media.

Affinity-purification mass spectrometry (AP-MS) is an established technique for identifying protein-protein interactions (PPIs). The basic technology involves immobilizing a high-specificity ligand to a solid-phase support (e.g., an agarose or magnetic bead) to pull down protein(s) of interest from cell lysates. Although these supports are engineered to minimize interactions with background protein, the conventional method recovers mostly nonspecific binders. The law of mass action for dilute solutions has taught us to use an excess of beads to capture all target proteins, especially weakly interacting ones. However, modern microbead technology presents a binding environment that is much different from a dilute solution. We describe a fluidic platform that captures and processes ultralow nanoliter quantities of magnetic particles, simultaneously increasing the efficiency of PPI detection and strongly suppressing nonspecific binding. We demonstrate the concept with synthetic mixtures of tagged protein and illustrate performance with a variety of AP-MS experiment types. These include a BioID experiment targeting lamin-A interactors from HeLa cells and pulldowns using GFP-tagged proteins associated with a double-strand DNA repair mechanism. We show that efficient extraction requires saturation of the solid-phase support and that <10 nL of beads is sufficient to generate comprehensive protein interaction maps.

Nutrition support and intervention preferences of cancer survivors.

INTRODUCTION: Ascertaining cancer survivors' specific needs and preferences regarding nutrition information delivery is important in developing and designing future health interventions in oncology. The present study aimed to explore the nutrition information needs of cancer survivors and their preferences regarding intervention delivery. METHODS: The present study is a mixed-methods study comprising a cross-sectional survey and focus groups. Participants were eligible for inclusion if they were aged ≥ 18 years, living in Ireland with a cancer diagnosis and had completed treatment at least 6 months previously. Cancer support centres promoted the online survey on social media. Focus groups were conducted with 20 individuals. RESULTS: The cohort (n = 56) was predominantly female (n = 50; 89.3%) breast cancer survivors (n = 42; 75%). Seven (12.5%) had received nutrition advice from a dietitian. Most were interested in receiving nutrition advice (n = 52; 92.9%), with variability in delivery mode. There was interest in improving sleep quality (n = 35; 62.5%), making positive lifestyle changes (n = 28; 50%) and accessing cancer-specific physical activity classes (n = 27; 48.2%). The best time to deliver information was throughout the cancer journey (n = 31; 55.4%). Time and motivation (n = 15; 26.8%) were the main barriers. Facilitators were keeping healthy (n = 42; 75%) and weight maintenance (n = 31; 55.4%). Four themes emerged from thematic analysis: lack of nutrition guidance, an abundance of misinformation, one size does not fit all and dietitians as the preferred source of advice. CONCLUSIONS: There is a desire for evidence-based nutrition advice and for dietitians to be integrated into cancer clinics and cancer survivorship care. This research guides future nutrition interventions to improve cancer survivorship outcomes.

Sources of nutrition advice and desired nutrition guidance in oncology care: Patient's perspectives.

BACKGROUND: The present study aimed to describe the proportion of cancer patients and survivors who receive nutrition advice, the sources of that advice and to explore the nutrition advice this cohort would like to have received. METHODS: A descriptive cross-sectional survey was completed by 211 cancer patients and survivors recruited through two hospitals in the North-West of Ireland. The survey consisted of open-ended and closed questions that aimed to explore the provision of and sources of nutrition advice and desired guidance in this cohort. Descriptive analysis was conducted using SPPS, while open-ended questions underwent thematic analysis. RESULTS: Respondents were mainly female (n = 133; 63%), aged 50-69 years old (n = 118; 56%), attending the outpatient department (n = 128; 60.7%) and < 5 years since diagnosis (n = 150; 71.7%). Breast (n = 69; 32.7%) was the most common cancer type. Respondents experienced a mean ± SD of 5 ± 3.1 nutrition impact symptoms. The most common was changes in taste/smell (n = 122; 57.8%). Although 53.6% (n = 113) reported being given advice, only 34.1% (n = 72) received this advice from a dietitian. The main sources of nutritional advice were friends and family. Respondents expressed their desire for nutrition advice from a dietitian and that this should be individualised, clear and practical. There was also a desire to avoid misinformation and uncertainty. CONCLUSIONS: The results of the present study can be used to help tailor nutrition support for this group. There is a need to ensure that patients and survivors consistently receive evidence-based advice tailored to their needs, cancer type and treatment stage.

A qualitative study into cancer survivors' relationship with nutrition post-cancer treatment.

In Ireland, one in every 25 individuals is living with and beyond cancer, placing survivorship care at the forefront of strategic planning and service requirements. Several unmet needs of cancer survivors were identified in the National Cancer Strategy 2017-2026. Further reports from national bodies have highlighted how these unmet needs, including those of a nutritional nature, have not been prioritised. In Ireland, little information is available on how cancer diagnosis changes cancer survivors' perceptions of food and nutrition. Thus, this study aimed to describe cancer survivors' relationship with nutrition. Survivors who lived on the Island of Ireland and were at least 6 months post-treatment were eligible. Five semi-structured focus groups and two individual interviews were conducted online (n = 20). Focus groups and interviews were analysed using thematic analysis. The cohort was mostly female breast cancer survivors (n = 12) with an average age of 51.3 ± 12 years. Emergent themes illustrated how a cancer diagnosis appears to motivate cancer survivors to immediate positive dietary changes. Many new dietary introductions were in line with cancer survivor recommendations, while dietary supplement use and some nonevidence-supported actions were revealed. A minority of participants continued their original dietary intake prior to their diagnosis. Participants reported the impact of treatment-related side effects on their relationship with nutrition. The participants emphasised a desire to receive dietary advice from health professionals while undergoing treatment and survivorship.

European Healthy Hydration Awareness Campaign for Dietitians.

Up to 50% of the adult population fail to meet the recommended total water intake despite the adverse health outcomes associated with chronic low fluid intake. Total fluid intake plays an important role in the energy and nutrient intake of individuals. Dietitians' role is to advise and guide the population toward a healthier diet. However, based on existing evidence, dietitians' current knowledge and practices regarding healthy hydration is an area for improvement. The European Healthy Hydration Awareness Campaign (EuHHAC) is an initiative between the European Federation of the Associations of Dietitians (EFAD) and the Hydration and Health Department of Danone Research, which aims to increase knowledge and awareness of healthy hydration and provide dietitians with evidence-based resources to use in practice. Since 2016, a series of tasks were undertaken by the EuHHAC group targeting dietitians across Europe. These tasks included an online survey addressing gaps in hydration knowledge, a webinar providing information on the terminology and impact of hydration on health, an interactive session addressing facilitators and barriers to healthy hydration, and a tutorial summarizing key hydration information and take-home messages. Dietitians were actively engaged in each aspect. Based on the feedback received, awareness of hydration was increased among dietitians. To further enhance understanding, an online Hydration Resource Center, including the resources of EuHHAC, will be created and uploaded on the EFAD website. Once completed an evaluation study will be undertaken to assess the usefulness of the resources in practice. The deliverables will be cascaded across Europe and worldwide via social media, conferences, and publications.

Nutrition education and leadership for improved clinical outcomes: training and supporting junior doctors to run 'Nutrition Awareness Weeks' in three NHS hospitals across England.

BACKGROUND: One in four adults are estimated to be at medium to high risk of malnutrition when screened using the 'Malnutrition Universal Screening Tool' upon admission to hospital in the United Kingdom. The Need for Nutrition Education/Education Programme (NNEdPro) Group was developed to address this issue and the Nutrition Education and Leadership for Improved Clinical Outcomes (NELICO) is a project within this group.The objective of NELICO was to assess whether an intensive training intervention combining clinical and public health nutrition, organisational management and leadership strategies, could equip junior doctors to contribute to improvement in nutrition awareness among healthcare professionals in the National Health Service in England. METHODS: Three junior doctors were self-selected from the NNEdPro Group original training. Each junior doctor recruited three additional team members to attend an intensive training weekend incorporating nutrition, change management and leadership. This equipped them to run nutrition awareness weeks in their respective hospitals. Knowledge, attitudes and practices were evaluated at baseline as well as one and four months post-training as a quality assurance measure. The number and type of educational events held, pre-awareness week Online Hospital Survey results, attendance and qualitative feedback from training sessions, effectiveness of dissemination methods such as awareness stalls, Hospital Nutrition Attitude Survey results and overall feedback were also used to determine impact. RESULTS: When the weighted average score for knowledge, attitudes and practices at baseline was compared with four months post-intervention scores, there was a significant increase in the overall score (p = 0.03). All three hospital teams conducted an effective nutrition awareness week, as determined by qualitative data collected from interviews and feedback from educational sessions. CONCLUSION: The NELICO project and its resulting nutrition awareness weeks were considered innovative in terms of concept and content. It was considered useful, both for the junior doctors who showed improvement in their nutrition knowledge and reported enthusiasm and for the hospital setting, increasing awareness of clinical and public health nutrition among healthcare professionals. The NELICO project is one innovative method to promote nutrition awareness in tomorrow's doctors and shows they have the enthusiasm and drive to be nutrition champions.

Qualitative Analysis Following the Delivery of a Farmer Lifestyle Program.

BACKGROUND: Irish farmers are adversely affected by noncommunicable diseases. Although there has been an increase in farmer health promotion activities in Ireland, farmers views on lifestyle programs are currently unknown. OBJECTIVES: To qualitatively analyze the impact of the previously mentioned 6-week physical activity and health education intervention on farmer health and to investigate how best to support this cohort moving forward. METHODS: A qualitative study was conducted online (two interviews, three focus groups) with fourteen Irish farmers (53.5 ± 6.5 years) who completed the 6-week program in December 2019. Interviews and focus groups were recorded, transcribed, and analyzed for themes. RESULTS: The main themes that emerged from this study were barriers, facilitators, and recommendations for lifestyle programs aimed to improve farmer health. Additional views on health and lifestyle behaviors were mentioned. Time of year was reported as the main barrier for farmers to engage in lifestyle programs. The key facilitators reported by farmers were the social health benefits obtained from the program and the farmer-specific nature of the program. Farmers suggested that physical activity and health education programs that are farmer-specific, delivered locally and catering for all fitness abilities should be more widely available to them. Although some farmers reported that they maintained the lifestyle behaviors they established during the initial 6-week program, follow-up supports are needed to encourage sustainable behavior change. CONCLUSIONS: Interventions that are farmer-specific, community-based, and feasible within the context of available resources may be effective in improving farmer health. Working in partnership with organizations that support farmers has the potential to improve farmer health.

Meaning of nutrition for cancer survivors: a photovoice study.

Background: Little is known about how cancer survivors perceive nutrition through the cancer experience and how those perceptions may influence their diet. Aims: This study aimed to capture the meaning of nutrition for cancer survivors who are post-cancer treatment using a participatory photography method known as photovoice. Methods: Wang and Burris's photovoice procedure was followed. Recruitment took place via email through existing links with participants from a previous quantitative study. The participants were tasked with taking photographs to represent the meaning of nutrition for them post-treatment. Group workshops and semistructured interviews were conducted to facilitate reflection, dialogue and analysis. Data analysis followed Braun and Clarke's six-phase thematic analysis. Results: One man and seven women (n=8) across the Island of Ireland were recruited. Participants identified six themes (illustrated with photographs): (1) Fresh is Best, (2) Be kind to yourself, (3) Building Blocks. Be Informed., (4) Post-Treatment Healing Changes, (5) Chemo Rituals and (6) Food for the Soul-Healthy Mind. Healthy Body. Conclusions: Participants displayed a holistic approach to a healthy lifestyle for recovery post-treatment and maintaining health. While diverse, participants made post-treatment nutritional changes by introducing and eliminating certain foods or food groups. All agreed that being informed and building nutrition knowledge are essential. It is important to clarify the implications cancer has had on diet and health when providing nutrition guidance to ensure that it is appropriate and specific.

Automating data analysis for hydrogen/deuterium exchange mass spectrometry using data-independent acquisition methodology.

We present a hydrogen/deuterium exchange workflow coupled to tandem mass spectrometry (HX-MS2) that supports the acquisition of peptide fragment ions alongside their peptide precursors. The approach enables true auto-curation of HX data by mining a rich set of deuterated fragments, generated by collisional-induced dissociation (CID), to simultaneously confirm the peptide ID and authenticate MS1-based deuteration calculations. The high redundancy provided by the fragments supports a confidence assessment of deuterium calculations using a combinatorial strategy. The approach requires data-independent acquisition (DIA) methods that are available on most MS platforms, making the switch to HX-MS2 straightforward. Importantly, we find that HX-DIA enables a proteomics-grade approach and wide-spread applications. Considerable time is saved through auto-curation and complex samples can now be characterized and at higher throughput. We illustrate these advantages in a drug binding analysis of the ultra-large protein kinase DNA-PKcs, isolated directly from mammalian cells.

Phosphorylation of polynucleotide kinase/ phosphatase by DNA-dependent protein kinase and ataxia-telangiectasia mutated regulates its association with sites of DNA damage.

Human polynucleotide kinase/phosphatase (PNKP) is a dual specificity 5'-DNA kinase/3'-DNA phosphatase, with roles in base excision repair, DNA single-strand break repair and non-homologous end joining (NHEJ); yet precisely how PNKP functions in the repair of DNA double strand breaks (DSBs) remains unclear. We demonstrate that PNKP is phosphorylated by the DNA-dependent protein kinase (DNA-PK) and ataxia-telangiectasia mutated (ATM) in vitro. The major phosphorylation site for both kinases was serine 114, with serine 126 being a minor site. Ionizing radiation (IR)-induced phosphorylation of cellular PNKP on S114 was ATM dependent, whereas phosphorylation of PNKP on S126 required both ATM and DNA-PK. Inactivation of DNA-PK and/or ATM led to reduced PNKP at DNA damage sites in vivo. Cells expressing PNKP with alanine or aspartic acid at serines 114 and 126 were modestly radiosensitive and IR enhanced the association of PNKP with XRCC4 and DNA ligase IV; however, this interaction was not affected by mutation of PNKP phosphorylation sites. Purified PNKP protein with mutation of serines 114 and 126 had decreased DNA kinase and DNA phosphatase activities and reduced affinity for DNA in vitro. Together, our results reveal that IR-induced phosphorylation of PNKP by ATM and DNA-PK regulates PNKP function at DSBs.

An exploration of self-perceived competence in providing nutrition care among physiotherapists in Ireland: a cross-sectional study.

BACKGROUND: Integrating nutrition care into physiotherapy can address modifiable risk factors contributing to chronic diseases, providing comprehensive and effective patient care, and supporting a wellness-oriented approach to healthcare. OBJECTIVE: To investigate physiotherapists' self-perceived confidence in their competence in nutrition care in Ireland. METHODS: Cross-sectional study using data from a validated online survey tool. Four constructs of competence in nutrition care were assessed: knowledge, skill, communication and counseling, and attitude. Open-ended responses were collected to gather opinions on nutrition knowledge requirements. Participants were chartered physiotherapists representing public and private workplaces across geographical settings in Ireland. RESULTS: 447 physiotherapists completed the survey. Most were female (n = 364, 81%), in private practice (n = 136, 31%), and located in a city (n = 215, 48%) with a mean 17 years post-qualification experience. Participants' self-perceived confidence in nutrition care competence was positive (mean score of 107.2/175). However, knowledge (mean score of 18.5/35) and skills (mean score of 27.2/55) related to nutrition care received lower ratings. Age and years of practice were positively associated with higher confidence in providing nutrition care. Most participants (n = 314, 71%) agreed that additional nutrition education is needed. Three overarching themes were identified regarding nutrition knowledge requirements: importance of providing nutrition advice, stated knowledge needs for nutrition education and training, and feasibility of nutrition care within physiotherapy practice. CONCLUSIONS: Physiotherapists in Ireland have confidence in provision of nutrition care but rated their knowledge and skills in nutrition as relatively low. Nutrition knowledge and skill are essential for physiotherapist practice as they can significantly impact patients' outcomes.

EASO and EFAD Position Statement on Medical Nutrition Therapy for the Management of Overweight and Obesity in Children and Adolescents.

INTRODUCTION: This position statement on medical nutrition therapy in the management of overweight or obesity in children and adolescents was prepared by an expert committee convened by the European Association for the Study of Obesity (EASO) and developed in collaboration with the European Federation of the Associations of Dietitians (EFAD). METHODS: It is based on the best evidence available from systematic reviews of randomized controlled trials on child and adolescent overweight and obesity treatment and other relevant peer-reviewed literature. RESULTS: Multicomponent behavioural interventions are generally considered to be the gold standard treatment for children and adolescents living with obesity. The evidence presented in this position statement confirms that dietary interventions can effectively improve adiposity-related outcomes. Dietary strategies should focus on the reduction of total energy intake through promotion of food-based guidelines that target modification of usual eating patterns and behaviours. These should target increasing intakes of nutrient-rich foods with a lower energy density, specifically vegetables and fruits, and a reduction in intakes of energy-dense nutrient-poor foods and beverages. In addition, higher intensity, longer duration treatments, delivered by interventionists with specialized dietetic-related skills and co-designed with families, are associated with greater treatment effects. DISCUSSION: Such interventions should be resourced adequately so that they can be implemented in a range of settings and in different formats, including digital or online delivery, to enhance accessibility.

European Association for the Study of Obesity Position Statement on Medical Nutrition Therapy for the Management of Overweight and Obesity in Adults Developed in Collaboration with the European Federation of the Associations of Dietitians.

INTRODUCTION: Obesity affects nearly 1 in 4 European adults increasing their risk for mortality and physical and psychological morbidity. Obesity is a chronic relapsing disease characterized by abnormal or excessive adiposity with risks to health. Medical nutrition therapy based on the latest scientific evidence should be offered to all Europeans living with obesity as part of obesity treatment interventions. METHODS: A systematic review was conducted to identify the latest evidence published in the November 2018-March 2021 period and to synthesize them in the European guidelines for medical nutrition therapy in adult obesity. RESULTS: Medical nutrition therapy should be administered by trained dietitians as part of a multidisciplinary team and should aim to achieve positive health outcomes, not solely weight changes. A diverse range of nutrition interventions are shown to be effective in the treatment of obesity and its comorbidities, and dietitians should consider all options and deliver personalized interventions. Although caloric restriction-based interventions are effective in promoting weight reduction, long-term adherence to behavioural changes may be better supported via alternative interventions based on eating patterns, food quality, and mindfulness. The Mediterranean diet, vegetarian diets, the Dietary Approaches to Stop Hypertension, portfolio diet, Nordic, and low-carbohydrate diets have all been associated with improvement in metabolic health with or without changes in body weight. In the November 2018-March 2021 period, the latest evidence published focused around intermittent fasting and meal replacements as obesity treatment options. Although the role of meal replacements is further strengthened by the new evidence, for intermittent fasting no evidence of significant advantage over and above continuous energy restriction was found. Pulses, fruit and vegetables, nuts, whole grains, and dairy foods are also important elements in the medical nutrition therapy of adult obesity. DISCUSSION: Any nutrition intervention should be based on a detailed nutritional assessment including an assessment of personal values, preferences, and social determinants of eating habits. Dietitians are expected to design interventions that are flexible and person centred. Approaches that avoid caloric restriction or detailed eating plans (non-dieting approaches) are also recommended for improvement of quality of life and body image perceptions.

Nutrition Practices among Adult Cancer Survivors Living on the Island of Ireland: A Cross-Sectional Study.

The purpose of this research was to explore the nutrition practices among post-treatment cancer survivors across Ireland. Cancer survivors aged 18+ years living across Ireland, who were not palliative and had completed active cancer treatment at least six months previous, were recruited to complete an online survey assessing dietary quality, food choice and satisfaction with food-related life as well as clinical and nutrition status. It was circulated by cancer support networks and on social media. Descriptive statistics are presented. The cohort (n = 170) was predominantly female (85.9%) and had breast cancer (64.7%). Mean age was 51.5 ± 10.9 years and 42.7% of the cohort were >five years post-treatment. Only 20% and 12% of the cohort had been assessed by a dietitian during and post-treatment, respectively. The mean dietary quality score was 10.3 ± 1.7, which was measured by the Leeds short-form food frequency questionnaire (SFFFQ). Using a 5-point Likert scale, the median satisfaction with food-related life score was 19 (3.3), which evaluates cognitive judgements on the person's food-related life. The food choice questionnaire (FCQ) assesses the relative importance of a range of factors related to dietary choice to individuals. The primary determinant of food choice in this cohort was the natural content (31.7%) followed by health (24.7%). Vitamin and mineral supplement use was reported by 69.8% of the cohort; the most consumed was Vitamin D. Four themes emerged from an optional open-ended question: awareness of nutritional importance; desire for specific nutritional advice and dietetic referral; cancer and treatment nutrition impacts were highlighted; as well as struggles with weight gain. This research provides useful insight into the nutrition practices of Irish cancer survivors. A desire and need for individualised and specific advice are evident.

An exploration of the self-perceived nutrition competencies of pharmacists.

Background: Pharmacists are integral members of healthcare teams, providing accessible nutrition care to patients, carers and health professionals. However, little is known about nutrition competence among pharmacists. Objective: The aim of the study is to determine the self-perceived competence in providing nutrition care among pharmacists in Ireland. Methods: This cross-sectional study used a mixed methods design consisting of an online survey delivered to pharmacists in Ireland. The pharmacists' self-perceived competencies according to confidence in knowledge, skills, communication and counselling, and attitudes in nutrition care were assessed using the validated NUTCOMP questionnaire. Qualitative responses in relation to nutrition knowledge were also gathered. Questionnaire responses were analysed using descriptive statistics and free text narratives were coded into themes and subthemes. Results: A total of n = 557 (74% (n = 413) female; 24.1% (n = 134) male) respondents completed the questionnaire providing a 14.9% response rate. Confidence in nutrition knowledge (mean score (SD) 21.6 (±4.31) out of a maximum of 35 points), nutrition skills (mean score (SD) 31.8 (±7.74) out of a maximum count of 55) and nutrition related counselling and communication (mean score (SD) 29.8 (±6.35) out of a maximum of 45 points) were limited. Most respondents had completed a programme with some nutrition content (60.8%) either formally or informally. Previous nutrition education was positively associated with greater scores in knowledge, skills, communication/counselling and attitudes towards nutrition care (p = ≤0.001). Over three-quarters of respondents (78.1%) agreed that they would like further nutrition education to support themselves in their roles as pharmacists. Thematic analysis highlights the role of interprofessional nutrition care, barriers to providing nutritional care in practice and opportunities for improving nutrition training. Conclusions: Most pharmacists reported being somewhat confident in their nutrition knowledge, skills and attitudes and are willing to participate in further nutrition education to support their patients. The findings suggest that there are other barriers to providing nutrition care in the pharmacy setting such as time, renumeration and lack of community dietitians.

Viral Manipulation of a Mechanoresponsive Signaling Axis Disassembles Processing Bodies.

Processing bodies (PBs) are ribonucleoprotein granules important for cytokine mRNA decay that are targeted for disassembly by many viruses. Kaposi's sarcoma-associated herpesvirus is the etiological agent of the inflammatory endothelial cancer, Kaposi's sarcoma, and a PB-regulating virus. The virus encodes kaposin B (KapB), which induces actin stress fibers (SFs) and cell spindling as well as PB disassembly. We now show that KapB-mediated PB disassembly requires actin rearrangements, RhoA effectors, and the mechanoresponsive transcription activator, YAP. Moreover, ectopic expression of active YAP or exposure of ECs to mechanical forces caused PB disassembly in the absence of KapB. We propose that the viral protein KapB activates a mechanoresponsive signaling axis and links changes in cell shape and cytoskeletal structures to enhanced inflammatory molecule expression using PB disassembly. Our work implies that cytoskeletal changes in other pathologies may similarly impact the inflammatory environment.

'Bhavishya Shakti: Empowering the Future': establishing and evaluating a pilot community mobile teaching kitchen as an innovative model, training marginalised women to become nutrition champions and culinary health educators in Kolkata, India.

BACKGROUND: Malnutrition is a global emergency, creating an overlapping burden on individual, public and economic health. The double burden of malnutrition affects approximately 2.3 billion adults worldwide. Following 3 years of capacity building work in Kolkata, with assistance of local volunteers and organisations, we established an empowering nutrition education model in the form of a 'mobile teaching kitchen (MTK)' with the aim of creating culinary health educators from lay slum-dwelling women. AIMS: To evaluate the piloting of a novel MTK nutrition education platform and its effects on the participants, alongside data collection feasibility. METHODS: Over 6 months, marginalised (RG Kar and Chetla slums) women underwent nutrition training using the MTK supported by dietitians, doctors and volunteers. Preintervention and postintervention assessments of knowledge, attitudes and practices (KAP), as well as anthropometric and clinical nutritional status of both the women and their children were recorded. The education was delivered by a 'See One, Do One, Teach One' approach with a final assessment of teaching delivery performed in the final session. RESULTS: Twelve women were trained in total, six from each slum. Statistically significant improvements were noted in sections of KAP, with improvements in nutrition knowledge (+4.8) and practices (+0.8). In addition, statistically significant positive changes were seen in 'understanding of healthy nutrition for their children' (p=0.02), 'sources of protein rich food' (p=0.02) and 'not skipping meals if a child is ill' (p≤0.001). CONCLUSION: The MTK as a public health intervention managed to educate, empower and upskill two groups of lay marginalised women into MTK Champions from the urban slums of Kolkata, India. Improvements in their nutrition KAP demonstrate just some of the effects of this programme. By the provision of healthy meals and nutritional messages, the MTK Champions are key drivers nudging improvements in nutrition and health related awareness with a ripple effect across the communities that they serve. There is potential to upscale and adapt this programme to other settings, or developing into a microenterprise model, that can help future MTK Champions earn a stable income.

Three-year review of a capacity building pilot for a sustainable regional network on food, nutrition and health systems education in India.

BACKGROUND: In Kolkata (India), there are high rates of malnourished children (45.9%) under the age of three, impacting growth, organ development, function, and cognition. Mothers have a major role to play during this crucial development stage, with research showing nutrition knowledge, attitudes and practices (KAP) of mothers are important determinants of childhood malnutrition. AIMS: To document 3 years of capacity building towards a sustainable nutrition education network in Kolkata, India, while assessing the ability to perform data collection in the form of needs assessments, impact assessments and capacity reviews. METHODS: Descriptive review and analysis of engagement and impact from 3 years of work by the NNEdPro Global Centre for Nutrition and Health, initiating locally led nutrition education interventions. Mapping to the Indian National Nutrition Strategy was also performed to review adherence to nationwide priorities surrounding nutrition and determine the wider application potential of the network. RESULTS: Two simultaneous projects were taken forward by a team of local healthcare professionals and student champions. Project 1-medical college workshops for medical student nutrition education with added focus on underserved populations, Project 2-preparation for a 'Mobile Teaching Kitchen' (MTK) in marginalised communities to empower local women as nutrition educators.Data collection methods used for analysing markers of impact and sustainability were semi-structured interviews of the community members, and KAP questionnaires to assess response to educational sessions. CONCLUSION: With local support it is possible to create and sustain fieldwork for an extended period with meaningful outputs and impact. This initiative demonstrates that it is possible to use healthcare professionals, students and volunteers with low-intensity training and a low-cost approach to produce action research with considerable impact and results in rapid, reliable and robust manner.

trans Autophosphorylation at DNA-dependent protein kinase's two major autophosphorylation site clusters facilitates end processing but not end joining.

Recent studies have established that DNA-dependent protein kinase (DNA-PK) undergoes a series of autophosphorylation events that facilitate successful completion of nonhomologous DNA end joining. Autophosphorylation at sites in two distinct clusters regulates DNA end access to DNA end-processing factors and to other DNA repair pathways. Autophosphorylation within the kinase's activation loop regulates kinase activity. Additional autophosphorylation events (as yet undefined) occur that mediate kinase dissociation. Here we provide the first evidence that autophosphorylation within the two major clusters (regulating end access) occurs in trans. Further, both UV-induced and double-strand break (DSB)-induced phosphorylation in the two major clusters is predominantly autophosphorylation. Finally, we show that while autophosphorylation in trans on one of two synapsed DNA-PK complexes facilitates appropriate end processing, this is not sufficient to promote efficient end joining. This suggests that end joining in living cells requires additional phosphorylation events that either occur in cis or that occur on both sides of the DNA-PK synapse. These data support an emerging consensus that, via a series of autophosphorylation events, DNA-PK undergoes a sequence of conformational changes that promote efficient and appropriate repair of DSBs.