Decoding the dynamics of cellular metabolism and the action of 3-bromopyruvate and 2-deoxyglucose using pulsed stable isotope-resolved metabolomics

BACKGROUND: Cellular metabolism is highly dynamic and continuously adjusts to the physiological program of the cell. The regulation of metabolism appears at all biological levels: (post-) transcriptional, (post-) translational, and allosteric. This regulatory information is expressed in the metabolome, but in a complex manner. To decode such complex information, new methods are needed in order to facilitate dynamic metabolic characterization at high resolution. RESULTS: Here, we describe pulsed stable isotope-resolved metabolomics (pSIRM) as a tool for the dynamic metabolic characterization of cellular metabolism. We have adapted gas chromatography-coupled mass spectrometric methods for metabolomic profiling and stable isotope-resolved metabolomics. In addition, we have improved robustness and reproducibility and implemented a strategy for the absolute quantification of metabolites. CONCLUSIONS: By way of examples, we have applied this methodology to characterize central carbon metabolism of a panel of cancer cell lines and to determine the mode of metabolic inhibition of glycolytic inhibitors in times ranging from minutes to hours. Using pSIRM, we observed that 2-deoxyglucose is a metabolic inhibitor, but does not directly act on the glycolytic cascade

Correlation between Endosonographic and Doppler Ultrasound Features of Portal Hypertension in Patients with Cirrhosis

Purpose. Endoscopic ultrasound (EUS) permits the detailed visualization of clinically significant features of portal hypertension; however, it is an invasive procedure that is not widely available. The aim of this cross-sectional study was to determine whether a correlation exists between the features of portal hypertension detected using both Doppler ultrasound and EUS in subjects with liver cirrhosis. Materials and Methods. Analyzed cohort included 42 patients who underwent a detailed Doppler ultrasound focusing on the parameters of blood flow in the portal/splenic vein as well as an endoscopic/EUS procedure that included the assessment of the size and localization of “deep” varices. Results. The size of “deep” oesophageal varices detected with EUS exhibited no correlation with the parameters assessed by Doppler ultrasound. However, the size of the “deep” gastric varices detected using EUS correlated with the time averaged maximum velocity (Tmax as well as Vmin, Vmax) for the portal vein using Doppler ultrasound and exhibited a correlation with the Vmax and Tmax for the splenic vein. No significant correlation was determined between the diameter of the azygous vein and the thickness of the gastric wall when seen on EUS versus the parameters measured with Doppler ultrasound. Conclusion. EUS provides important information regarding the features of portal hypertension, and in the case of “deep” oesophageal varices exhibits a limited correlation with the parameters detected by Doppler ultrasound. Thus, despite its invasiveness, EUS is a method that provides a reliable and unique assessment of the features of portal hypertension in patients with liver cirrhosis

Barriers and facilitators to uptake and retention of inner-city ethnically diverse women in a postnatal weight management intervention: a mixed-methods process evaluation within a feasibility trial in England.

OBJECTIVES: To understand the barriers and facilitators to uptake and retention of postnatal women randomised to a commercial group weight management intervention using the COM-B (capability, opportunity, motivation and behaviour) behaviour change model. DESIGN: Concurrent mixed-methods (qualitative dominant) process evaluation nested within a feasibility randomised controlled trial, comprising questionnaires and interviews at 6 and 12 months postbirth. SETTING: One National Health Service maternity unit in an inner city area in the south of England. PARTICIPANTS: 98 postnatal women with body mass indices>25 kg/m2 (overweight/obese) at pregnancy commencement. INTERVENTION: Twelve-week Slimming World (SW) commercial group weight management programme, commencing anytime from 8 to 16 weeks postnatally. PRIMARY AND SECONDARY OUTCOME MEASURES: Data regarding uptake and retention from questionnaires and interviews conducted 6 and 12 months postbirth analysed thematically and mapped to the COM-B model. RESULTS: Barriers to SW uptake mostly concerned opportunity issues (eg, lack of time or childcare support) though some women also lacked motivation, not feeling that weight reduction was a priority, and a few cited capability issues such as lacking confidence. Weight loss aspirations were also a key factor explaining retention, as were social opportunity issues, particularly in relation to factors such as the extent of group identity and relationship with the group consultant; and physical opportunity such as perceived support from and fit with family lifestyle. In addition, barriers relating to beliefs and expectations about the SW programme were identified, including concerns regarding compatibility with breastfeeding and importance of exercise. Women's understanding of the SW approach, and capability to implement into their lifestyles, appeared related to level of attendance (dose-response effect). CONCLUSIONS: Uptake and retention in commercial weight management programmes may be enhanced by applying behaviour change techniques to address the barriers impacting on women's perceived capability, motivation and opportunity to participate. TRIAL REGISTRATION NUMBER: ISRCTN39186148

Agricultural landscapes as multi-scale public good and the role of the Common Agricultural Policy

During the last 50 years, the Common Agricultural Policy (CAP) has impacted the evolution of European agricultural landscapes by driving changes in land use and farming practices. We propose a typology characterizing the scales relevant for agricultural landscapes management and argue that action is required on three scales: (1) a landscape oriented management at farm level, (2) the coordination of land managers\u27 actions at landscape level, and (3) the conservation of the diversity of agricultural landscapes in the EU. We provide evidence that the CAP has until now mainly focused on the first scale. We also illustrate how agricultural policy could encourage coordinated actions at the landscape- and EU-scales. In particular, we propose policy instruments to coordinate actions of individual land owners (e.g. collective bonus in agro-environmental contracts or support to environmental cooperatives). We also analyse how the recognition and transposition of the European Landscape Convention could promote trans-frontier landscape cooperation in order, not only to conserve high-quality rural landscapes, but also to ensure the conservation of the diversity of EU landscapes (scale 3). This article provides a knowledge base to support an integrated CAP design in the direction of improved landscape management, as an important component of the EU project towards more sustainable agriculture

Motion analysis of elite Polish soccer goalkeepers throughout a season

The study aims were to determine the distance covered by goalkeepers during matches in the context of game duration and result, to identify the area of their most frequent activity, and to assess goalkeepers' involvement in games finished with a win, draw, or loss. The investigation was based on two innovative tools: the goalkeeper's activity index (GAI) and an analysis of 5-min periods. A video tracking system was used to monitor 17 goalkeepers from Polish National League teams during 15 matches. The GAI was applied to assess their involvement in the game. Elite goalkeepers covered 72.7%, 25.8%, and 2.5% of the distance during the game by walking/jogging, running, and sprinting, respectively. The distances covered in lost, won, and drawn matches turned out similar (mean \ub1 SD: 4800 \ub1 906 m, 4696 \ub1 1033 m, and 4660 \ub1 754 m, respectively). There were no significant differences between the distances covered in the first and second halves. The area of most frequent activity was the middle sector of the penalty area between the goal and penalty area lines. ANOVA results showed that in drawn matches, goalkeepers' activity significantly differed in mean values of the GAI in comparison with that in won and lost games (p = 0.034, p = 0.039, respectively). It was noted that goalkeepers tended to intervene more often in games where their team was winning rather than in those with a losing result. Their direct involvement in defending the goal was the lowest in drawn games

Increasing role of arthropod bites in tularaemia transmission in Poland – case reports and diagnostic methods

The study describes four cases of tularaemia – one developed after contact with rabbits and three developed after an arthropod bite. Due to non-specific clinical symptoms, accurate diagnosis of tularaemia may be difficult. The increasing contribution of the arthropod vectors in the transmission of the disease indicates that special effort should be made to apply sensitive and specific diagnostic methods for tularaemia, and to remind health-care workers about this route of Francisella tularensis infections. The advantages and disadvantages of various diagnostic methods – molecular, serological and microbiological culture – are discussed. The PCR as a rapid and proper diagnostic method for ulceroglandular tularaemia is presented

Preplant 1,3-D treatments test well for perennial crop nurseries, but challenges remain

Preplant fumigation with methyl bromide commonly is used in open-field perennial crop nurseries in California for control of plant-parasitic nematodes, pathogens and weeds. Because this fumigant is being phased out, alternatives are needed to ensure the productivity of the perennial crop nursery industry as well as the ornamental, orchard and vineyard production systems that depend on clean planting stock. As part of the USDA Area-Wide Pest Management Program for Integrated Methyl Bromide Alternatives, several perennial crop nursery projects were conducted in California from 2007 to 2011 to test and demonstrate registered alternative fumigants and application techniques that maximize performance and minimize environmental impacts. The project was designed to evaluate shank application and soil surface sealing methods intended to reduce aboveground emission and improve soil performance of 1,3-dichloropropene, a leading methyl bromide alternative for nurseries. In these garden rose and tree nursery experiments, 1,3-dichloropropene treatments performed well regardless of application techniques. In this article, we highlight recent research and discuss the significance and remaining challenges for adoption of methyl bromide alternatives in this unique nursery stock production system

Alterations of mTOR signaling impact metabolic stress resistance in colorectal carcinomas with BRAF and KRAS mutations

Metabolic reprogramming is as a hallmark of cancer, and several studies have reported that BRAF and KRAS tumors may be accompanied by a deregulation of cellular metabolism. We investigated how BRAF(V600E) and KRAS(G12V) affect cell metabolism, stress resistance and signaling in colorectal carcinoma cells driven by these mutations. KRAS(G12V) expressing cells are characterized by the induction of glycolysis, accumulation of lactic acid and sensitivity to glycolytic inhibition. Notably mathematical modelling confirmed the critical role of MCT1 designating the survival of KRAS(G12V) cells. Carcinoma cells harboring BRAF(V600E) remain resistant towards alterations of glucose supply or application of signaling or metabolic inhibitors. Altogether these data demonstrate that an oncogene-specific decoupling of mTOR from AMPK or AKT signaling accounts for alterations of resistance mechanisms and metabolic phenotypes. Indeed the inhibition of mTOR in BRAF(V600E) cells counteracts the metabolic predisposition and demonstrates mTOR as a potential target in BRAF(V600E)-driven colorectal carcinomas