Enzymatic synthesis of novel fructosylated compounds by Ffase from Schwanniomyces occidentalis in green solvents

The β-fructofuranosidase from the yeast Schwanniomyces occidentalis (Ffase) produces potential prebiotic fructooligosaccharides (FOS) by self-transfructosylation of sucrose, being one of the highest known producers of 6-kestose. The use of Green Solvents (GS) in biocatalysis has emerged as a sustainable alternative to conventional organic media for improving product yields and generating new molecules. In this work, the Ffase hydrolytic and transfructosylating activity was analysed using different GS, including biosolvents and ionic liquids. Among them, 11 were compatible for the net synthesis of FOS. Besides, two glycerol derivatives improved the yield of total FOS. Interestingly, polyols ethylene glycol and glycerol were found to be efficient alternative fructosyl-acceptors, both substantially decreasing the sucrose fructosylation. The main transfer product of the reaction with glycerol was a 62 g L−1 isomeric mixture of 1-O and 2-O-β-D-fructofuranosylglycerol, representing 95% of all chemicals generated by transfructosylation. Unexpectedly, the non-terminal 2-O fructo-conjugate was the major molecule catalysed during the process, while the 1-O isomer was the minor one. This fact made Ffase the first known enzyme from yeast showing this catalytic ability. Thus, novel fructosylated compounds with potential applications in food, cosmetics, and pharmaceutical fields have been obtained in this work, increasing the biotechnological interest of Ffase with innocuous GS

Knowledge, attitudes, behavioral and organizational factors of health professions students for a competent smoking cessation practice: An instrument adaptation and psychometric validation study in Spanish and English samples

Background: To improve smoking cessation, training of health professions students is essential. However, no specific instrument is available to assess factors that may affect students' learning about smoking cessation practice. Aim: To adapt and validate the Knowledge, Attitudes, Behaviors and Organization questionnaire in the population of undergraduate health professions students. Design: Methodological research. Methods: The researchers conducted this study with 511 Spanish and 186 English health professions students from four different universities. We used a four-step approach: 1) adaptation of the items to the target population and validation of the content by a panel of experts; 2) a pilot study to test face validity; 3) linguistic adaptation of the Spanish version to English; and 4) the psychometric assessment based on construct validity, criterion validity and internal consistency. Results: Exploratory factor analysis revealed four subscales for the Spanish version, namely 'Individual knowledge and skills', 'Individual attitudes and beliefs', 'Organizational support' and 'Organizational resources', which accounted for 85.1% of the variance. Confirmatory factor analysis in the holdout Spanish and English samples revealed adequate goodness-of-fit values, supporting the factor structure. Hypotheses testing demonstrated significant differences by capacitation in smoking cessation interventions and degree courses, providing further evidence regarding construct validity. All the subscales correlated positively with the criterion variables (5 A's smoking cessation model), except for the 'Organizational resources' subscale, which was not significantly correlated with the 5 A's. The overall Cronbach's alpha was.83 for the Spanish version and.88 for the English one. Conclusions: Our results provide empirical support for the use of the Knowledge, Attitudes, Behaviors and Organization questionnaire for Students as a reliable and valid instrument to assess knowledge, attitudes, behaviors and organization perceptions in health professions students, which is essential for competent smoking cessation practice. Interestingly, 'Organizational resources' subscale presented the lowest correlations among factors and did not correlate with any component of the 5 A's, suggesting the need of enhancing students' responsibility and involvement during their internships, as well as the interest of some organizations

G protein-specific mechanisms in the serotonin 5-HT2A receptor regulate psychosis-related effects and memory deficits

G protein-coupled receptors (GPCRs) are sophisticated signaling machines able to simultaneously elicit multiple intracellular signaling pathways upon activation. Complete (in)activation of all pathways can be counterproductive for specific therapeutic applications. This is the case for the serotonin 2 A receptor (5-HT2AR), a prominent target for the treatment of schizophrenia. In this study, we elucidate the complex 5-HT2AR coupling signature in response to different signaling probes, and its physiological consequences by combining computational modeling, in vitro and in vivo experiments with human postmortem brain studies. We show how chemical modification of the endogenous agonist serotonin dramatically impacts the G protein coupling profile of the 5-HT2AR and the associated behavioral responses. Importantly, among these responses, we demonstrate that memory deficits are regulated by Gαq protein activation, whereas psychosis-related behavior is modulated through Gαi1 stimulation. These findings emphasize the complexity of GPCR pharmacology and physiology and open the path to designing improved therapeutics for the treatment of stchizophrenia.This work was supported by the ERAnet NEURON consortium fund (funding was provided by CIHR NDD-161471 and FRQ-S 278647 for M.B., the German Federal Ministry of Education and Research under grant number 01EW1909 for P.K., as well as the Instituto de Salud Carlos III and Fondo Europeo de Desarrollo Regional number AC18/00030 for J.S. and P.R.). This work was further supported by the Instituto de Salud Carlos III (ISCIII) and co-funded by the European Union (PI18/00094) to J.S. and (PI18/00053) to P.R. We acknowledge grant support from Agencia Estatal de Investigación (PID2020-119428RB-I00; SAF2017-88126R), Basque Government (IT-1211/19, IT-1512/22 and KK-2019/00-49), Xunta de Galicia (ED431C 2022/20 and ED431G 2019/02) and European Regional Development Fund (ERDF). P.K. thanks the German Research Foundation DFG for Heisenberg Professorship KO4095/5-1. S.S.O. and R.S. thank the PTQ-17-09103 (Ayuda Torres Quevedo, Ministerio de Ciencia e Innovación), and BioExcel-2 (Grant Number 823830, Horizon2020). M.B. was in part supported by an operating grant (# PJT 183758) from the Canadian Institute for Health Research. I.M-A. was the recipient of a predoctoral fellowship from the Basque Government. The authors would like to thank the staff members of the Basque Institute of Legal Medicine for their cooperation in the study, especially to Dr. Benito Morentin. R.D-A., T.M.S, D.A.G., I.M.A., A.S., P.K. and J.S. are members of COST Action CA18133 “ERNEST”

Dietary inflammatory index and all-cause mortality in large cohorts: The SUN and PREDIMED studies

[Background]: Inflammation is known to be related to the leading causes of death including cardiovascular disease, several types of cancer, obesity, type 2 diabetes, depression-suicide and other chronic diseases. In the context of whole dietary patterns, the Dietary Inflammatory Index (DII®) was developed to appraise the inflammatory potential of the diet. [Objective]: We prospectively assessed the association between DII scores and all-cause mortality in two large Spanish cohorts and valuated the consistency of findings across these two cohorts and results published based on other cohorts.[Design]: We assessed 18,566 participants in the “Seguimiento Universidad de Navarra” (SUN) cohort followed-up during 188,891 person-years and 6790 participants in the “PREvencion con DIeta MEDiterránea” (PREDIMED) randomized trial representing 30,233 person-years of follow-up. DII scores were calculated in both cohorts from validated FFQs. Higher DII scores corresponded to more proinflammatory diets. A total of 230 and 302 deaths occurred in SUN and PREDIMED, respectively. In a random-effect meta-analysis we included 12 prospective studies (SUN, PREDIMED and 10 additional studies) that assessed the association between DII scores and all-cause mortality.[Results]: After adjusting for a wide array of potential confounders, the comparison between extreme quartiles of the DII showed a positive and significant association with all-cause mortality in both the SUN (hazard ratio [HR] = 1.85; 95% CI: 1.15, 2.98; P-trend = 0.004) and the PREDIMED cohort (HR = 1.42; 95% CI: 1.00, 2.02; P-trend = 0.009). In the meta-analysis of 12 cohorts, the DII was significantly associated with an increase of 23% in all-cause mortality (95% CI: 16%–32%, for the highest vs lowest category of DII).[Conclusion]: Our results provide strong and consistent support for the hypothesis that a pro-inflammatory diet is associated with increased all-cause mortality. The SUN cohort and PREDIMED trial were registered at clinicaltrials.gov as NCT02669602 and at isrctn.com as ISRCTN35739639, respectively.Supported by the official funding agency for biomedical research of the Spanish Government, Instituto de Salud Carlos III (ISCIII), through grants provided to research networks specifically developed for the trial (RTIC G03/140, to R.E.; RTIC RD 06/0045, to Miguel A. Martínez-González) and through Centro de Investigación Biomédica en Red de Fisiopatología de la Obesidad y Nutrición (CIBERobn), and by grants from Centro Nacional de Investigaciones Cardiovasculares (CNIC 06/2007), Fondo de Investigación Sanitaria–Fondo Europeo de Desarrollo Regional (Proyecto de Investigación (PI) 04-2239, PI 05/2584, CP06/00100, PI07/0240, PI07/1138, PI07/0954, PI 07/0473, PI10/01407, PI10/02658, PI11/01647, P11/02505, PI13/00462, PI13/00615, PI13/01090, PI14/01668, PI14/01798, PI14/01764), Ministerio de Ciencia e Innovación (Recursos y teconologia agroalimentarias(AGL)-2009-13906-C02 and AGL2010-22319-C03 and AGL2013-49083-C3-1- R), Fundación Mapfre 2010, the Consejería de Salud de la Junta de Andalucía (PI0105/2007), the Public Health Division of the Department of Health of the Autonomous Government of Catalonia, Generalitat Valenciana (Generalitat Valenciana Ayuda Complementaria (GVACOMP) 06109, GVACOMP2010-181, GVACOMP2011-151), Conselleria de Sanitat y, PI14/01764 AP; Atención Primaria (CS) 2010-AP-111, and CS2011-AP-042), and Regional Government of Navarra (P27/2011).). Drs. Shivappa and Hébert were supported by grant number R44DK103377 from the United States National Institute of Diabetes and Digestive and Kidney Diseases

Dietary α-linolenic acid, marine ω-3 fatty acids, and mortality in a population with high fish consumption: Findings from the PREvención con DIeta MEDiterránea (PREDIMED) Study

12 Páginas.-- 6 Tablas.-- 1 FiguraBackground-Epidemiological evidence suggests a cardioprotective role of α-linolenic acid (ALA), a plant-derived ω-3 fatty acid. It is unclear whether ALA is beneficial in a background of high marine ω-3 fatty acids (long-chain n-3 polyunsaturated fatty acids) intake. In persons at high cardiovascular risk from Spain, a country in which fish consumption is customarily high, we investigated whether meeting the International Society for the Study of Fatty Acids and Lipids recommendation for dietary ALA (0.7% of total energy) at baseline was related to all-cause and cardiovascular disease mortality. We also examined the effect of meeting the society's recommendation for long-chain n-3 polyunsaturated fatty acids (≥500 mg/day). Methods and Results-We longitudinally evaluated 7202 participants in the PREvención con DIeta MEDiterránea (PREDIMED) trial. Multivariable-adjusted Cox regressionmodels were fitted to estimate hazard ratios. ALA intake correlated towalnut consumption (r=0.94). During a 5.9-y follow-up, 431 deaths occurred (104 cardiovascular disease, 55 coronary heart disease, 32 sudden cardiac death, 25 stroke). The hazard ratios formeeting ALArecommendation (n=1615, 22.4%) were 0.72 (95% CI 0.56-0.92) for all-causemortality and 0.95 (95% CI 0.58-1.57) for fatal cardiovascular disease. The hazard ratios formeeting the recommendation for long-chain n-3 polyunsaturated fatty acids (n=5452, 75.7%) were 0.84 (95% CI 0.67-1.05) for all-causemortality, 0.61 (95% CI 0.39-0.96) for fatal cardiovascular disease, 0.54 (95% CI 0.29-0.99) for fatal coronary heart disease, and 0.49 (95% CI 0.22-1.01) for sudden cardiac death. The highest reduction in all-cause mortality occurred in participants meeting both recommendations (hazard ratio 0.63 [95% CI 0.45-0.87]). Conclusions-In participants without prior cardiovascular disease and high fish consumption, dietary ALA, supplied mainly by walnuts and olive oil, relates inversely to all-cause mortality, whereas protection from cardiac mortality is limited to fish-derived long-chain n-3 polyunsaturated fatty acids.This study was funded in part by Instituto de Salud Carlos III (ISCIII) (Spanish Ministry of Economy) through grants RTIC G03/140, RTIC RD 06/0045, Centro Nacional de Investigaciones Cardiovasculares CNIC 06/2007, ISCIII FIS PS09/01292, the Spanish Ministry of Science and Innovation (MICINN) AGL2010‐22319‐C03‐02 and AGL2009‐13906‐C02‐02, and an unrestricted grant from the California Walnut Commission. Sala‐Vila holds a Miguel Servet I fellowship from the Ministry of Economy and Competitiveness through the ISCIII