Complex metaprobe production and characterisation for metabolic studies in relation to bowel health

Introduction - Colorectal cancer is the third most common cancer in the UK and the second most common cause of cancer mortality. There is increasing evidence that the presence of a systemic inflammatory response plays an important role in predicting survival in patients with colorectal cancer. The Glasgow Prognostic Score includes a measure of systemic inflammatory response and has been used successfully as a predictor of survival rate in cancer patients. Butyric acid is one of the short chain fatty acids (SCFA) produced by colonic bacteria through fermentation of dietary non-digestible carbohydrate (NDC). This simple organic acid is known to have anti-inflammatory and possibly direct anti-neoplastic effects. The inaccessibility of the human colon means that little is known of the in vivo rate of butyrate production, its metabolic fate or the foodstuffs that may maximise its production. Butyrate may be produced in greater quantities from other SCFA such as acetate and lactate rather than by direct production by saccharolytic bacteria. Specialist organisms may be responsible for producing butyrate through conversion of other SCFA which are produced by fermentation in the colon. The distal site of butyrate production may be a very important feature and the implication that slowly fermentable NDC, such as resistant starch, may be significant. Previous Work - The European Union project EUROSTARCH (www.eurostarch.org) investigated the way in which different starchy foods are digested and metabolised within the body. The results of this project have informed the development of low GI foods. Results have also increased our knowledge of NDC fermentation and promise to lead to improvements in human bowel health. We pioneered the measurement of starch digestion and fermentation during this project. What are Metaprobes? The term metaprobe is used to describe stable isotope labelled tracers. Complex metaprobes are produced by biological synthesis e.g. the introduction of 13C enriched CO2 gas during the production of crops. Simple metaprobes are produced by chemical synthesis e.g. urea. We can produce complex metaprobes by labelling staple foods such as cereals using the stable isotope 13C incorporated from enriched 13CO2 during anthesis of the crops. Complex metaprobes are unique tools with which to study human metabolism in health and disease and they can be used in several ways. The labelled cereals allow in vivo measurement of butyrate production from dietary NDC and to permit its production to be monitored. Hypothesis - 30% of cancer deaths in the UK are bowel related. Butyric acid produced naturally through fermentation of dietary carbohydrates, is a potent and natural anti-inflammatory agent which can protect against cancer. We urgently need to develop methods to quantify butyrate production in vivo. Aims - Three aims are recognised: optimising the preparation of complex metaprobes of barley and wheat grain; optimising yield and 13C-enrichment of whole grain and its starch and protein fractions; using complex metaprobes to compare SCFA production in vivo. Plan of Work - This project will include a review of complex metaprobe production for human nutrition and health studies in relation to bowel health. The practical work will include production and characterisation of complex metaprobes (13C-labelled cereals) and their application to study butyrate production in a pilot study on volunteer human subjects. I have successfully grown cereal crops with different varieties of barley and wheat in glasshouse conditions. During the anthesis phase, the plants were placed in growth cabinets at 18oC with 18 hours of light in every 24 hours. The plants were labelled using 13C enriched CO2 gas and are incubated for a further 24 hours and returned to the glasshouse. Examples of the harvest are Wheat var Paragon produced 1.7 Kg labelled grain at 0.278 ape 13C enrichment and Barley var Cellar produced 3.4 Kg labelled grain at 0.198 ape 13C enrichment. Some of the 13C labelled grain has already been used for human nutrition studies. The 13C-enrichment of starch glucose has been measured by liquid chromatography isotope ratio mass spectrometry (LC-IRMS) analysis. These cereal grains can be incorporated into common foodstuffs for in vivo studies of the production of butyrate and other SCFA. Volunteers were fed two meals made from 13C-enriched barley grain: porridge and whole grain barley. Whole grains being rich in resistant starch. Samples of urine will be used to measure butyrate production. Breath samples will be used to measure oxidation of the 13C-enriched barley, which is a global measure of digestion and fermentation. Laboratory procedures such as ultrafiltration and solid phase extraction will be applied to urine samples. These sample preparation procedures will be followed by SCFA analysis by GC-IRMS. Future Direction - Future clinical trials will be considered with Prof Donald C McMillan, University Dept of Surgery, Glasgow Royal Infirmary as a means of identifying the role of butyrate in colonic cancer. The current screening process recognises three phases: early stage diagnosis; polyp development; diagnosis and treatment of advance stage colonic cancer. We wish to study the role of butyrate and its production using our newly developed complex metaprobe tools

Plant organellar RNA maturation

Plant organellar RNA metabolism is run by a multitude of nucleus-encoded RNA-binding proteins (RBPs) that control RNA stability, processing, and degradation. In chloroplasts and mitochondria, these post-transcriptional processes are vital for the production of a small number of essential components of the photosynthetic and respiratory machinery—and consequently for organellar biogenesis and plant survival. Many organellar RBPs have been functionally assigned to individual steps in RNA maturation, often specific to selected transcripts. While the catalog of factors identified is ever-growing, our knowledge of how they achieve their functions mechanistically is far from complete. This review summarizes the current knowledge of plant organellar RNA metabolism taking an RBP-centric approach and focusing on mechanistic aspects of RBP functions and the kinetics of the processes they are involved in.Peer Reviewe

Gender, Research, and Advocacy: What We’ve Learned and What Work Lies Ahead

From the global Women’s Marches to the #metoo movement and SDG 5, women’s rights and gender equality are clearly among the most urgent issues for the international human rights movement today. Long-held commitments and established language on women’s rights and gender equality are under threat at all levels of governance and in issue areas ranging from sexual violence to reproductive health to political participation. If history has made one thing clear, it’s that women’s rights are fragile and constantly at risk of being extinguished for political purposes. This panel will explore the following four themes from the conference in light of this global reality: Connecting Scholars and Practitioners: What are the opportunities and limitations of connecting research and advocacy when working on women’s rights and/or gender equality issues? Which research methods are ideal for bridging scholars and practitioners on these issues? Movements: To what extent do we or should we distinguish between studying (and doing) women’s activism and using a gender perspective in human rights advocacy? Are there ways in which greater visibility to women’s issues has narrowed our understandings of women? To the exclusion of other gender-related issues? Technology: What role does technology play in creating visibility for gender issues and voices? For sharing research on human rights advocacy or aiding mobilization? Neoliberal Solutions: What is the status of philanthropy and funding for work on gender issues? Have market-based solutions to raise funds for NGOs become dominant in ways that perpetuate stereotypes

Explaining participation in the informal economy a purchaser perspective

Purpose Participation in the informal economy has been predominantly explained from a supply-side perspective by evaluating the rationales for people working in this sphere. Recognising that many transactions in the informal economy are often instigated by customers, exemplified by purchasers asking “how much for cash?”, the aim of this paper is to explain the informal economy from a demand-side perspective by evaluating citizens‟ rationales for making purchases in the informal economy. Here, we test three potential explanations for acquiring goods and services in the informal economy, grounded in rational economic actor, social actor and formal economy imperfections theoretical perspectives. Methodology To do this, a 2013 Eurobarometer survey, involving 27,563 face-to-face interviews conducted in 28 European Union member states is reported. Findings The finding is that all three rationales apply but the weight given to each varies across populations. A multinomial logit regression analysis then pinpoints the specific groups variously using the informal economy to obtain a lower price, for social or redistributive rationales, or due to the failures of the formal economy in terms of the availability, speed and quality of provision. Practical Implications The outcome is to reveal that the policy approach of changing the cost/benefit ratios confronting purchasers will only be effective for those purchasers citing a lower price as their prime rationale. Different policy measures will be required for those making informal economy purchases due to the shortcomings of the formal economy, and for social ends. These policy measures are then discussed. Originality/value The value and originality of this paper is that it explains participation in the informal economy from a purchaser, rather than the predominant supplier, perspective

Time-dependent genetic effects on gene expression implicate aging processes

Gene expression is dependent on genetic and environmental factors. In the last decade, a large body of research has significantly improved our understanding of the genetic architecture of gene expression. However, it remains unclear whether genetic effects on gene expression remain stable over time. Here, we show, using longitudinal whole-blood gene expression data from a twin cohort, that the genetic architecture of a subset of genes is unstable over time. In addition, we identified 2213 genes differentially expressed across time points that we linked with aging within and across studies. Interestingly, we discovered that most differentially expressed genes were affected by a subset of 77 putative causal genes. Finally, we observed that putative causal genes and down-regulated genes were affected by a loss of genetic control between time points. Taken together, our data suggest that instability in the genetic architecture of a subset of genes could lead to widespread effects on the transcriptome with an aging signature

Fat and Carbohydrate Interact to Potentiate Food Reward in Healthy Weight but Not in Overweight or Obesity

Prior work suggests that actual, but not estimated, energy density drives the reinforcing value of food and that energy from fat and carbohydrate can interact to potentiate reward. Here we sought to replicate these findings in an American sample and to determine if the effects are influenced by body mass index (BMI). Thirty participants with healthy weight (HW; BMI 21.92 ± 1.77; M ± SD) and 30 participants with overweight/obesity (OW/OB; BMI 29.42 ± 4.44) rated pictures of common American snacks in 120-kcal portions for liking, familiarity, frequency of consumption, expected satiety, healthiness, energy content, energy density, and price. Participants then completed an auction task where they bid for the opportunity to consume each food. Snacks contained either primarily carbohydrate, primarily fat, or roughly equal portions of fat and carbohydrate (combo). Replicating prior work, we found that participants with HW bid the most for combo foods in linear mixed model analyses. This effect was not observed among individuals with OW/OB. Additionally, in contrast with previous reports, our linear regression analyses revealed a negative relationship between the actual energy density of the snacks and bid amount that was mediated by food price. Our findings support altered macronutrient reinforcement in obesity and highlight potential influences of the food environment on the regulation of food reward

Hospital-Based Acute Care Use in Survivors of Septic Shock

OBJECTIVES: Septic shock is associated with increased long-term morbidity and mortality. However, little is known about the use of hospital-based acute care in survivors after hospital discharge. The objectives of the study were to examine the frequency, timing, causes, and risk factors associated with emergency department visits and hospital readmissions within 30 days of discharge. DESIGN: Retrospective cohort study. SETTING: Tertiary, academic hospital in the United States. PATIENTS: Patients admitted with septic shock (serum lactate ≥ 4 mmol/L or refractory hypotension) and discharged alive to a nonhospice setting between 2007 and 2010. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The coprimary outcomes were all-cause hospital readmission and emergency department visits (treat-and-release encounters) within 30 days to any of the three health system hospitals. Of 269 at-risk survivors, 63 (23.4%; 95% CI, 18.2-28.5) were readmitted within 30 days of discharge and another 12 (4.5%; 95% CI, 2.3-7.7) returned to the emergency department for a treat-and-release visit. Readmissions occurred within 15 days of discharge in 75% of cases and were more likely in oncology patients (p=0.001) and patients with a longer hospital length of stay (p=0.04). Readmissions were frequently due to another life-threatening condition and resulted in death or discharge to hospice in 16% of cases. The reasons for readmission were deemed potentially related to the index septic shock hospitalization in 78% (49 of 63) of cases. The most common cause was infection related, accounting for 46% of all 30-day readmissions, followed by cardiovascular or thromboembolic events (18%). CONCLUSIONS: The use of hospital-based acute care appeared to be common in septic shock survivors. Encounters often led to readmission within 15 days of discharge, were frequently due to another acute condition, and appeared to result in substantial morbidity and mortality. Given the potential public health implications of these findings, validation studies are needed

Developing laboratory capacity for Good Laboratory Practice certification: lessons from a Tanzanian insecticide testing facility.

Background: With increasing insecticide resistance in malaria-endemic countries there is an urgent need for safe and effective novel vector control products. To improve the capacity of facilities that test insecticides in sub-Saharan Africa, a programme is supporting seven facilities towards Good Laboratory Practice (GLP) certification, the globally recognized standard for quality management system (QMS) for the conduct of non-clinical and environmental studies. The World Health Organization (WHO) GLP Handbook provides guidance on a stepwise approach to implement a GLP compliant QMS. This study assesses auditor GLP checklists and timings outlined in the WHO GLP Handbook in the real-life context of a Tanzanian insecticide-testing facility, evaluating their implementation in this context. Methods and Principle Findings: We conducted document review and semi-structured interviews with staff at all levels of the test facility to explore factors that influenced progress towards GLP certification. We found that while auditor GLP checklists underemphasised computer systems, they were otherwise broadly applicable. Factors that delayed time to completion of GLP certification included the need for extensive infrastructure improvements, the availability of regional expertise related to GLP, the capacity of national and regional external systems and services to meet GLP compliance requirements, and training development required for Standard Operating Procedure implementation. Conclusion: The standards required for full GLP compliance are rigorous, with an expected completion timeline to implementation of 24 months. This study shows that in low and middle-income countries this timeline may be unrealistic due to challenges related to infrastructure development and lack of regional capacity and expertise. We recommend a comprehensive gap analysis when starting a project, including these areas which are beyond those recommended by the WHO GLP Handbook. These challenges can be successfully overcome and the experience in Tanzania provides key lessons for other facilities seeking GLP certification or the development of similar QMS

First Cultivation and Characterization of Mycobacterium ulcerans from the Environment

Mycobacterium ulcerans infection, or Buruli ulcer, is the third most common mycobacteriosis of humans worldwide, after tuberculosis and leprosy. Buruli ulcer is a neglected, devastating, necrotizing disease, sometimes producing massive, disfiguring ulcers, with huge social impact. Buruli ulcer occurs predominantly in impoverished, humid, tropical, rural areas of Africa, where the incidence has been increasing, surpassing tuberculosis and leprosy in some regions. Besides being a disease of the poor, Buruli ulcer is a poverty-promoting chronic infectious disease. There is strong evidence that M. ulcerans is not transmitted person to person but is an environmental pathogen transmitted to humans from its aquatic niches. However, until now M. ulcerans has not been isolated in pure culture from environmental sources. This manuscript describes the first isolation, to our knowledge, of M. ulcerans in pure culture from an environmental source. This strain, which is highly virulent for mice, has microbiological features typical of African strains of M. ulcerans and was isolated from an aquatic insect from a Buruli ulcer–endemic area in Benin, West Africa. Our findings support the concept that M. ulcerans is a pathogen of humans with an aquatic environmental niche and will have positive consequences for the control of this neglected and socially important tropical disease