Isolation of Extracellular Vesicles from Phloem Sap by Size Exclusion Chromatography.

Extracellular vesicles (EVs) are nanoparticles that are released by cells and participate in the transfer of information. It is now known that EVs from mammalian cells are involved in different physiological and pathophysiological processes (antigen presentation, tissue regeneration, cancer, inflammation, diabetes, etc.). In the past few years, several studies on plants have demonstrated that EVs are also key tools for plant intercellular and cross-kingdom communications, suggesting that these nanostructures may contribute to distinct aspects of plant physiology such as development, defense, reproduction, symbiotic relationships, etc. These findings are challenging the traditional view of signaling in plants. EVs are probably involved in the phloem's transport system, since this vascular tissue plays a crucial role in translocating nutrients, defensive compounds, and informational signals throughout the plant. The collection of phloem is experimentally challenging because sap is under high turgor pressure inside the sieve elements, which have a small diameter and are hidden within the plant organs. The goals of this work are to develop new protocols that allow us to detect EVs for the first time in the phloem of the plants, and to isolate these nanovesicles for in-depth analysis and characterization. Our protocols describe two distinct methods to collect the phloem sap from rice and melon. The first method (Basic Protocol 1) involves 'Aphid stylectomy by radiofrequency microcautery' using rice plants and the aphid Sitobion avenae. This is considered the least invasive method for collecting phloem sap. The second method, 'Stem incision', involves cutting the stem of melon plants for collecting the exuded sap. Phloem sap EVs are then isolated by size exclusion chromatography. The results obtained in this study represent the first report on typical EVs isolated from in vivo-collected phloem sap. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Isolation of EVs from phloem sap: Aphid stylectomy by radiofrequency microcautery Basic Protocol 2: Isolation of EVs from phloem sap: Stem incision method.

Extracellular vesicles from the trematodes Fasciola hepatica and Dicrocoelium dendriticum trigger different responses in human THP-1 macrophages.

Extracellular vesicles (EVs) released by the helminths Dicrocoelium dendriticum and Fasciola hepatica are important modulators of the host immune response, contributing to the establishment of the infection. Monocytes and, in particular, macrophages are major regulators of the inflammatory response and are likely responsible for the phagocytosis of most of the parasite EVs. In this study, we isolated EVs from F. hepatica (FhEVs) and D. dendriticum (DdEVs) by size exclusion chromatography (SEC) and characterized them by nanoparticle tracking analysis, transmission electron microscopy and LC-MS/MS, and analyzed the cohort of proteins. The treatment of monocytes/macrophages with FhEVs, DdEVs or EV-depleted fractions from SEC, demonstrated species-specific effects of the EVs. In particular, FhEVs reduce the migratory capacity of monocytes and the analysis of the cytokine profile showed that they induce a mixed M1/M2 response, exerting anti-inflammatory properties in Lipopolysaccharide-activated macrophages. In contrast, DdEVs do not affect monocyte migration and seem to have pro-inflammatory properties. These results correlate with the differences in the life cycle of both parasites, suggesting different host immune responses. Only F. hepatica migrates to the bile duct through the liver parenchyma, driving the host immune response to heal deep erosions. Furthermore, the proteomic analysis of the macrophages upon FhEV treatment identified several proteins that might be involved in FhEV-macrophage interactions.

Dietary-Derived Exosome-like Nanoparticles as Bacterial Modulators: Beyond MicroRNAs.

There is increasing evidence that food is an important factor that influences the composition of the gut microbiota. Usually, all the attention has been focused on nutrients such as lipids, proteins, vitamins, or polyphenols. However, a pivotal role in these processes has been linked to dietary-derived exosome-like nanoparticles (DELNs). While food macro- and micronutrient composition are largely well established, there is considerable interest in these DELNs and their cargoes. In this sense, traditionally, all the attention was focused on the proteins or miRNAs contained in these vesicles. However, it has been shown that DELNs would also carry other bioactive molecules with a key role in regulating biochemical pathways and/or interactions with the host's gut microbiome affecting intracellular communication. Due to the scarce literature, it is necessary to compile the current knowledge about the antimicrobial capacity of DELNs and its possible molecular mechanisms that will serve as a starting point. For this reason, in this review, we highlight the impact of DENLs on different bacteria species modulating the host gut microbiota or antibacterial properties. It could be concluded that DELNs, isolated from both plant and animal foods, exert gut microbiota modulation. However, the presence of miRNA in the vesicle cargoes is not the only one responsible for this effect. Lipids present in the DELNs membrane or small molecules packed in may also be responsible for apoptosis signaling, inhibition, or growth promoters.

'Nutritional Footprint' in the Food, Meals and HoReCa Sectors: A Review.

Nowadays, the food industry is integrating environmental, social, and health parameters to increase its sustainable impact. To do this, they are using new tools to calculate the potential efficiency of nutritional products with lower levels of environmental impact. One of these tools is called the 'nutritional footprint', created by Wuppertal Institute for Climate, Environment and Energy GmbH. This study aims to review this concept and clarify its historical development, its use in several sectors of the food industry, and its transformation from a manual to an online tool. Results reflected that it is a suitable indicator that integrates nutritional, environmental, and social-economic dimensions to help the decision-making process in the procurement of more sustainable products and, although it is limited to Germany due to the use of the national standard nutritional intakes of Germany, its importance lies in the fact that is a promising instrument to promote environmental sustainability in the context of food, meals, and the hotel, restaurant and catering (HoReCa) sectors.

Characterization and bioactivity of extracellular vesicles isolated from pomegranate.

In the current study, extracellular vesicles from pomegranate juice (PgEVs) were isolated for the first time using size exclusion chromatography (SEC). This method permitted us to obtain highly enriched EV samples without most of the non-EV co-isolated proteins. The characterization of PgEVs through nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM) allowed the determination of vesicles' concentration/volume, size, and morphology. It was confirmed from the analytical data that PgEVs contain a homogeneous population of vesicles, with a dimension and structure comparable to plant-derived EVs. Proteomic analyses by LC-MS/MS led to the characterization of 131 proteins, and several of them were related commonly to the biogenesis and transport of EVs, and/or proposed as EV markers. PgEVs exerted anti-inflammatory, antioxidant and wound-healing effects when added to the in vitro cultures of monocytic (THP-1) and intestinal (Caco-2) cell lines, respectively.

"Five Keys to Safer Food" and COVID-19.

On 11 March 2020, coronavirus disease 2019 (COVID-19) was declared a pandemic by the World Health Organization (WHO) and, up to 18:37 a.m. on 9 December 2021, it has produced 268,440,530 cases and 5,299,511 deaths. This disease, in some patients, included pneumonia and shortness of breath, being transmitted through droplets and aerosols. To date, there is no scientific literature to justify transmission directly from foods. In this review, we applied the precautionary principle for the home and the food industry using the known "Five Keys to Safer Food" manual developed by the World Health Organization (WHO) and extended punctually in its core information from five keys, in the light of new COVID-19 evidence, to guarantee a possible food safety tool.

COVID Obesity: A One-Year Narrative Review.

On 11 March 2020, coronavirus disease 2019 (COVID-19) was declared a pandemic by the World Health Organization (WHO). This study focuses on a narrative review about the illness during the first year of the pandemic in relation to obesity. Databases were used to search studies published up to 8 December 2020. In total, 4430 articles and other scientific literature were found, and 24 articles were included in this one-year narrative review. The mean BMI value of severe COVID-19 patients ranged from 24.5 to 33.4 kg/m2, versus <18.5 to 24.3 kg/m2 for non-severe patients. Articles using the terms obesity or overweight without indicating the BMI value in these patients were common, but this is not useful, as the anthropometric parameters, when not defined by this index, are confusing due to the classification being different in the West compared to among Asian and Korean criteria-based adults. We proposed a new term, called COVID obesity, to define the importance of this anthropometric parameter, among others, in relation with this pandemic.

An Internet-based treatment for Flying Phobia using 360° images: Study protocol for a feasibility pilot study.

BACKGROUND: Flying Phobia (FP) is a prevalent disorder that can cause serious interference in a person's life. ICBT interventions have already shown their efficacy in several studies, but studies in the field of specific phobias are still scarce. Moreover, few studies have investigated the feasibility of using different types of images in exposure scenarios in ICBTs and no studies have been carried out on the role of sense of presence and reality judgement. The aim of the present study is to explore the feasibility of an ICBT for FP (NO-FEAR Airlines) using two types of images with different levels of immersion (still and navigable images). A secondary aim is to explore the potential effectiveness of the two experimental conditions using two types of images compared to a waiting list control group. Finally, the role of navigable images compared to the still images in the level of anxiety, sense of presence, and reality judgement will also be explored. This paper presents the study protocol. METHODS: This study is a three-armed feasibility pilot study with the following conditions: NO-FEAR Airlines with navigable images, NO-FEAR Airlines with still images, and a waiting list group. A minimum of 60 participants will be recruited. The intervention will have a maximum duration of 6 weeks. Measurements will be taken at four different moments: baseline, post-intervention, and two follow-ups (3- and 12-month). Participants' opinions, preference, satisfaction and acceptance regarding the images used in the exposure scenarios will be assessed. FP symptomatology outcomes will also be considered for secondary analyses. The anxiety, sense of presence, and reality judgement in the exposure scenarios will also be analysed. DISCUSSION: This study will conduct a pilot study on the feasibility of an ICBT for FP and it is the first one to explore the evaluation of patients of the two type of images (still and navigable) and the role of presence and reality judgement in exposure scenarios delivered through the Internet. Research in this field can have an impact on the way these scenarios are designed and developed, as well as helping to explore whether they have any effect on adherence. TRIAL REGISTRATION: NCT03900559. Trial Registration date 3 April 2019, retrospectively registered.

Evolutionary Trends in the Mitochondrial Genome of Archaeplastida: How Does the GC Bias Affect the Transition from Water to Land?

Among the most intriguing mysteries in the evolutionary biology of photosynthetic organisms are the genesis and consequences of the dramatic increase in the mitochondrial and nuclear genome sizes, together with the concomitant evolution of the three genetic compartments, particularly during the transition from water to land. To clarify the evolutionary trends in the mitochondrial genome of Archaeplastida, we analyzed the sequences from 37 complete genomes. Therefore, we utilized mitochondrial, plastidial and nuclear ribosomal DNA molecular markers on 100 species of Streptophyta for each subunit. Hierarchical models of sequence evolution were fitted to test the heterogeneity in the base composition. The best resulting phylogenies were used for reconstructing the ancestral Guanine-Cytosine (GC) content and equilibrium GC frequency (GC*) using non-homogeneous and non-stationary models fitted with a maximum likelihood approach. The mitochondrial genome length was strongly related to repetitive sequences across Archaeplastida evolution; however, the length seemed not to be linked to the other studied variables, as different lineages showed diverse evolutionary patterns. In contrast, Streptophyta exhibited a powerful positive relationship between the GC content, non-coding DNA, and repetitive sequences, while the evolution of Chlorophyta reflected a strong positive linear relationship between the genome length and the number of genes.

Exploring the Relationship Between the Acceptability of an Internet-Based Intervention for Depression in Primary Care and Clinical Outcomes: Secondary Analysis of a Randomized Controlled Trial.

Background: Depression is one of the most prevalent psychological disorders worldwide. Although psychotherapy for depression is effective, there are barriers to its implementation in primary care in Spain. The use of the Internet has been shown to be a feasible solution. However, the acceptability of Internet-based interventions has not been studied sufficiently. Objective: To assess the acceptability of an Internet-based intervention (IBI) for depression in primary care, and explore the relationship between expectations and satisfaction and the improvement in the clinical variables in primary care patients receiving this intervention. Furthermore, it offers data about the effects of some sociodemographic characteristics on these acceptability variables and analyzes whether the expectations are related to finalizing the intervention. Methods: Data were based on depressive patients who were participants in a randomized controlled trial. In the present study, we present the data from all the participants in the Internet intervention groups (N = 198). All the participants filled out the expectation and satisfaction scales (six-item scales regarding treatment logic, satisfaction, recommending, usefulness for other disorders, usefulness for the patient, and unpleasantness), the Beck Depression Inventory-II, and the secondary outcome measures: depression and anxiety impairment, and positive and negative affect. Results: Results showed that participants' expectations and satisfaction with the program were both high and differences in expectations and satisfaction depended on some sociodemographic variables (age: older people have higher expectations; sex: women have greater satisfaction). A positive relationship between these variables and intervention efficacy was found: expectations related to "usefulness for the patient" were a statistically related predictor to the results on the BDI-II (Beta = 0.364), and the perception of how logical the treatment is (Beta = 0.528) was associated with change in the clinical variable. Furthermore, the higher the expectations, the higher the improvements exhibited by the patients in all measures evaluated during the ten intervention modules. High expectations were also directly related to finalizing the intervention. Conclusions: This is the first study in Spain to address this issue in the field of IBIs for depression in primary care. The IBI showed high acceptance related to the intervention's efficacy and completion. Research on IBI acceptability could help to implement the treatment offered. Clinical Trial Registration: www.ClinicalTrials.gov, identifier NCT01611818.

Prevalencia de hipertensión arterial y su asociación con antropometría y dieta en niños (de seis a nueve años): estudio ANIVA / Prevalence of arterial hypertension and its association with anthropometry and diet in children (6 to 9 years old): ANIVA study

Introducción: estudios recientes alertan de cambios en los hábitos alimentarios y de un aumento del sobrepeso en niños como posibles factores de riesgo de una hipertensión temprana. Objetivos: conocer la prevalencia de normotensión, prehipertensión e hipertensión en los niños estudiados y valorar su posible asociación con la dieta e indicadores antropométricos. Métodos: se realizó un estudio transversal dentro del Proyecto "Antropometría y Nutrición Infantil de Valencia" (ANIVA), donde se valoró dieta, presión arterial y antropometría en 652 niños/as de entre seis y nueve años. Se aplicaron los criterios de la Organización Mundial de la Salud (OMS) para identificar normotensos, prehipertensos e hipertensos. Resultados: existe una mayor prevalencia de normotensos (83,9%, p < 0,05). La prevalencia de prehipertensos e hipertensos es de un 8,1% y 8,0%, respectivamente. Los niños presentan mayores niveles de hipertensión (8,4%), mientras que las niñas presentan mayor prevalencia de prehipertensión (9,3%). En los niños se identifican valores significativamente mayores en el peso, la altura y la circunferencia de cadera. Las niñas presentan signifi cativamente mayor porcentaje de masa grasa, pliegue bicipital, pliegue suprailiaco y frecuencia cardiaca. Se encontraron correlaciones significativas entre los indicadores antropométricos y la presión arterial por sexo, destacando la correlación perímetro de cintura-presión arterial en los niños. Se observan diferencias significativas entre los nutrientes ingeridos en la dieta y la presión arterial. Conclusiones: se identifica una prevalencia de hipertensión del 8% en niños de 6-9 años de la provincia de Valencia. Los valores de presión arterial aumentan en función del índice de masa corporal, el porcentaje de masa grasa, el perímetro de cintura y el conjunto de los pliegues. Se identifi ca un patrón dietético asociado a la hipertensión en niños

Introduction: recent studies warn of changes in dietary habits and an increase in overweight children as possible risk factors for early hypertension. Objectives: to know the prevalence of normotension, prehypertension and hypertension in the children studied, and to evaluate its possible association with diet and anthropometric indicators. Methods: a cross-sectional study was carried out within the Project "Anthropometry and Child Nutrition of Valencia" (ANIVA), where diet, blood pressure, and anthropometry were evaluated in 652 children between six and nine years old. The World Health Organization (WHO) criteria to identify normotensive, prehypertensive and hypertensive were applied. Results: there is a higher prevalence of normotensive (83.9%, p < 0.05). The prevalence of prehypertensive and hypertensive patients is 8.1% and 8.0%, respectively. Boys have higher levels of hypertension (8.4%), while girls have a higher prevalence of prehypertension (9.3%). In boys, significantly higher values are identified for the weight, height and hip circumference. Girls present significantly higher percentage of fat mass, bicipital fold, suprailiac fold, and heart rate. Significant correlations were found between the anthropometric indicators and blood pressure by sex, highlighting the waist circumference-blood pressure correlation in boys. Significant differences are observed between the nutrients ingested in the diet and blood pressure. Conclusions: a prevalence of hypertension of 8% in children aged 6-9 years of the province of Valencia is identified. The blood pressure values increase according to the body mass index, percentage of fat mass, waist circumference and the set of folds. A dietary pattern associated with hypertension in children is identified

Assessing Chronic Stress, Coping Skills, and Mood Disorders through Speech Analysis: A Self-Assessment 'Voice App' for Laptops, Tablets, and Smartphones.

BACKGROUND: Computerized speech analysis (CSA) is a powerful method that allows one to assess stress-induced mood disturbances and affective disorders through repeated measurements of speaking behavior and voice sound characteristics. Over the past decades CSA has been successfully used in the clinical context to monitor the transition from 'affectively disturbed' to 'normal' among psychiatric patients under treatment. This project, by contrast, aimed to extend the CSA method in such a way that the transition from 'normal' to 'affected' can be detected among subjects of the general population through 10-20 self-assessments. METHODS: Central to the project was a normative speech study of 5 major languages (English, French, German, Italian, and Spanish). Each language comprised 120 subjects stratified according to gender, age, and education with repeated assessments at 14-day intervals (total n = 697). In a first step, we developed a multivariate model to assess affective state and stress-induced bodily reactions through speaking behavior and voice sound characteristics. Secondly, we determined language-, gender-, and age-specific thresholds that draw a line between 'natural fluctuations' and 'significant changes'. Thirdly, we implemented the model along with the underlying methods and normative data in a self-assessment 'voice app' for laptops, tablets, and smartphones. Finally, a longitudinal self-assessment study of 36 subjects was carried out over 14 days to test the performance of the CSA method in home environments. RESULTS: The data showed that speaking behavior and voice sound characteristics can be quantified in a reproducible and language-independent way. Gender and age explained 15-35% of the observed variance, whereas the educational level had a relatively small effect in the range of 1-3%. The self-assessment 'voice app' was realized in modular form so that additional languages can simply be 'plugged in' once the respective normative data become available. Results of the longitudinal self-assessment study in home environments demonstrated that CSA methods work well under most circumstances. CONCLUSIONS: We have successfully developed and tested a self-assessment CSA method that can monitor transitions from 'normal' to 'affected' in subjects of the general population in the broader context of mood disorders. Our easy-to-use 'voice app' evaluates sequences of 10-20 repeated assessments and watches for affect- and stress-induced deviations from baseline that exceed language-, gender-, and age-specific thresholds. Specifically, the 'voice app' provides users with stress-related 'biofeedback' and can help to identify that 10-15% subgroup of the general population that exhibits insufficient coping skills under chronic stress and may benefit from early detection and intervention prior to developing clinically relevant symptoms.

An Internet-based program for depression using activity and physiological sensors: efficacy, expectations, satisfaction, and ease of use.

PURPOSE: Computerized cognitive behavioral therapy (CCBT) has been shown to be efficacious. Moreover, CCBT can be enhanced by using physiological and activity sensors, but there is no evidence about the acceptability of all these tools. The objective of this study is to examine the efficacy, expectations, satisfaction, and ease of use of an Internet-based CCBT program for preventing depression, with and without sensors (electroencephalography, electrocardiograhpy ECG, and actigraphy), in a high-risk population (unemployed men). PATIENTS AND METHODS: Sixty participants at risk of depression (unemployed men) were randomly assigned to three experimental conditions: 1) intervention program (N=22), 2) intervention program plus sensors (N=19), and 3) control group (N=19). Participants completed depression, anxiety, positive and negative affect, and perceived stress measures. Furthermore, they also completed the measures for expectation, satisfaction, and the ease of use of the program. RESULTS: Results showed that the two intervention groups improved significantly more than the control group on the clinical variables, and the improvements were greater in the group that used sensors than in the group that did not use them. Furthermore, participants in both intervention groups scored high on expectations and satisfaction with the CCBT program (with and without sensors). The mean score for usability was 88 out of 100 (standard deviation =12.32). No significant differences were found between groups on any of these variables. CONCLUSION: This is the first study to analyze the efficacy, expectations, satisfaction, and ease of use of an Internet-based program using physiological and activity sensors. These results suggest that an Internet program for depression with or without physiological and activity sensors is effective, satisfactory, and easy to use.

Presence of microorganisms from isolated Megaselia spp. in foodservice establishments.

INTRODUCTION: The transmission of harmful pathogens by arthropods is an increasing health concern. More concretely, flies are known to be able to transmit the infectious agent mechanically. OBJECTIVE: The present work shows a case report occurred from foodservice establishments where were isolated and identified, at the first time, Megaselia spp. in the food preparation place. Furthermore, microorganisms were analyzed from these flies. METHOD: It is based in entomological and microbiological analysis. RESULTS: Mesophilic aerobic flora and Enterobacteriaceae were found in all the samples, exceeding the limits established from food commodities on 41.7% (5/12) for mesophilic aerobic bacteria and 66.7% (8/12) for Enterobacteriaceae. Furthermore, 25% (3/12) of analyzed flies were found positive to Escherichia coli, data that can be linked with the microbiological food results. The most surprising results were the presence of S. aureus in 66.7% (8/12) of the analyzed flies. CONCLUSIONS: A binomial relationship among Megaselia spp. and bacteria is demonstrated being an important study to demonstrate that must be checked more hygienically measurement in foodservice.

Introducción: la transmisión de patógenos por insectos es una creciente preocupación para la salud pública. Más concretamente, las moscas son conocidas por ser capaces de transmitir el agente infeccioso mecánicamente. Objetivo: el presente trabajo muestra un estudio en los servicios de restauración en los que se aisló por primera vez en la literatura Megaselia spp, detectándose la presencia de microorganismos en estas moscas. Método: se basa en análisis microbiológicos y entomológicos. Resultados: la presencia de aerobios mesófilos y Enterobacteriaceae se han encontrado en todas las muestras, superando los límites establecidos en el 41,7% (5/12) para las bacterias aerobias mesófilas y el 66,7% (8/12) para Enterobacteriaceae. Por otra parte, en el 25 y 66,7% de las moscas analizadas se detectó la presencia de Escherichia coli y Staphylococcus aureus, respectivamente. Conclusiones: hay un binomio entre la presencia de microorganismos y Megaselia spp., lo que demuestra la importancia de mantener una vigilancia más estricta en las medidas higiénico-sanitarias en los servicios de restauración.

Presence of microorganisms from isolated Megaselia spp. In foodservice establishments / Presencia de microorganismos aislados de Megaselia spp. En los establecimientos de servicios de restauración

Introduction: the transmission of harmful pathogens by arthropods is an increasing health concern. More concretely, flies are known to be able to transmit the infectious agent mechanically. Objective: the present work shows a case report occurred from foodservice establishments where were isolated and identified, at the first time, Megaselia spp. in the food preparation place. Furthermore, microorganisms were analyzed from these flies. Method: it is based in entomological and microbiological analysis. Results: mesophilic aerobic flora and Enterobacteriaceae were found in all the samples, exceeding the limits established from food commodities on 41.7% (5/12) for mesophilic aerobic bacteria and 66.7% (8/12) for Enterobacteriaceae. Furthermore, 25% (3/12) of analyzed flies were found positive to Escherichia coli, data that can be linked with the microbiological food results. The most surprising results were the presence of S. aureus in 66.7% (8/12) of the analyzed flies. Conclusions: a binomial relationship among Megaselia spp. and bacteria is demonstrated being an important study to demonstrate that must be checked more hygienically measurement in foodservice (AU)

Introducción: la transmisión de patógenos por insectos es una creciente preocupación para la salud pública. Más concretamente, las moscas son conocidas por ser capaces de transmitir el agente infeccioso mecánicamente. Objetivo: el presente trabajo muestra un estudio en los servicios de restauración en los que se aisló por primera vez en la literatura Megaselia spp, detectándose la presencia de microorganismos en estas moscas. Método: se basa en análisis microbiológicos y entomológicos. Resultados: la presencia de aerobios mesófilos y Enterobacteriaceae se han encontrado en todas las muestras, superando los límites establecidos en el 41,7% (5/12) para las bacterias aerobias mesófilas y el 66,7% (8/12) para Enterobacteriaceae. Por otra parte, en el 25 y 66,7% de las moscas analizadas se detectó la presencia de Escherichia coli y Staphylococcus aureus, respectivamente. Conclusiones: hay un binomio entre la presencia de microorganismos y Megaselia spp., lo que demuestra la importancia de mantener una vigilancia más estricta en las medidas higiénico-sanitarias en los servicios de restauración (AU)

Quantifying insufficient coping behavior under chronic stress: a cross-cultural study of 1,303 students from Italy, Spain and Argentina.

The question of how to quantify insufficient coping behavior under chronic stress is of major clinical relevance. In fact, chronic stress increasingly dominates modern work conditions and can affect nearly every system of the human body, as suggested by physical, cognitive, affective and behavioral symptoms. Since freshmen students experience constantly high levels of stress due to tight schedules and frequent examinations, we carried out a 3-center study of 1,303 students from Italy, Spain and Argentina in order to develop socioculturally independent means for quantifying coping behavior. The data analysis relied on 2 self-report questionnaires: the Coping Strategies Inventory (COPE) for the assessment of coping behavior and the Zurich Health Questionnaire which assesses consumption behavior and general health dimensions. A neural network approach was used to determine the structural properties inherent in the COPE instrument. Our analyses revealed 2 highly stable, socioculturally independent scales that reflected basic coping behavior in terms of the personality traits activity-passivity and defeatism-resilience. This replicated previous results based on Swiss and US-American data. The percentage of students exhibiting insufficient coping behavior was very similar across the study sites (11.5-18.0%). Given their stability and validity, the newly developed scales enable the quantification of basic coping behavior in a cost-efficient and reliable way, thus clearing the way for the early detection of subjects with insufficient coping skills under chronic stress who may be at risk of physical or mental health problems.

Propuesta de valores de referencia ambientales microbiológicos en los servicios de restauración / Proposal of reference values of microbiological environment monitoring in foodservice establishments

Introducción: Proponer una nueva referencia microbiológica ambiental en los servicios de restauración. Objetivo: El presente trabajo muestra la determinación y evaluación de la contaminación microbiológica generada en un establecimiento de servicio de alimentos. Método: Se basa en el muestreo microbiológico de superficie utilizando filtros de membrana de ésteres mixtos de celulosa y en el muestreo de aire utilizando placas de Petri. Resultados: Los límites de contaminación se establecieron antes y durante la elaboración de alimentos, mediante el análisis microbiológico de las superficies y del medio ambiente, sistemas de equipos, hasta límites confiables y estableciendo niveles de aceptación en cada punto seleccionado. Por último, se estableció un programa de vigilancia microbiológica del medio ambiente incluyendo la evaluación de todos los parámetros que la componen y están implícitos en la zona, asegurando de esta manera y el apoyo a su continuidad con la documentación y los registros desarrollados para el área de seguridad. Las muestras para el análisis microbiológico se recogieron durante un período durante diez días diferentes, en dos horas diferentes. Se identificaron doce puntos como peligrosos. Además, los alimentos clasificados como de alto riesgo fueron recogidos periódicamente para el análisis microbiológico. Conclusiones: La posibilidad de utilizar de una amplia gama de medios selectivos, muy por encima del número limitado utilizado en este estudio, permite el análisis de muchas especies microbianas individuales (AU)

Introduction: To propose a new reference of microbiological environment monitoring in foodservice establishments. Objective: The present work shows the determination and evaluation of the microbiological contamination generated in a foodservice establishment Method: It is based on surface sampling (microbial build-up) using mixed cellulose ester membrane filters and on air sampling (hourly microbial adhesion) using Petri dishes. Results: Limits of contamination are established before and during the food elaboration, by means of the microbiological analysis of the environment, surfaces and equipment systems, until reliable limits and levels of acceptance are established of each selected point. Finally, a program of environmental microbiological monitoring was established including the evaluation of all parameters that compose and are implicit in the area, thus assuring and supporting its continuity with the documentation and registers developed for a safety area. Samples for microbiological examination were collected over a period of one moth on ten different days, at two different times. Twelve selected points having previously been identified as hazardous were monitored. Furthermore, foods though to be of high risk were periodically collected for microbiological analysis. Conclusions: The possibility to use of an ample range of selective media, well over the limited number used in this study, allows the analysis of many single microbial species (AU)

Affective state and voice: cross-cultural assessment of speaking behavior and voice sound characteristics--a normative multicenter study of 577 + 36 healthy subjects.

BACKGROUND: Human speech is greatly influenced by the speakers' affective state, such as sadness, happiness, grief, guilt, fear, anger, aggression, faintheartedness, shame, sexual arousal, love, amongst others. Attentive listeners discover a lot about the affective state of their dialog partners with no great effort, and without having to talk about it explicitly during a conversation or on the phone. On the other hand, speech dysfunctions, such as slow, delayed or monotonous speech, are prominent features of affective disorders. METHODS: This project was comprised of four studies with healthy volunteers from Bristol (English: n = 117), Lausanne (French: n = 128), Zurich (German: n = 208), and Valencia (Spanish: n = 124). All samples were stratified according to gender, age, and education. The specific study design with different types of spoken text along with repeated assessments at 14-day intervals allowed us to estimate the 'natural' variation of speech parameters over time, and to analyze the sensitivity of speech parameters with respect to form and content of spoken text. Additionally, our project included a longitudinal self-assessment study with university students from Zurich (n = 18) and unemployed adults from Valencia (n = 18) in order to test the feasibility of the speech analysis method in home environments. RESULTS: The normative data showed that speaking behavior and voice sound characteristics can be quantified in a reproducible and language-independent way. The high resolution of the method was verified by a computerized assignment of speech parameter patterns to languages at a success rate of 90%, while the correct assignment to texts was 70%. In the longitudinal self-assessment study we calculated individual 'baselines' for each test person along with deviations thereof. The significance of such deviations was assessed through the normative reference data. CONCLUSIONS: Our data provided gender-, age-, and language-specific thresholds that allow one to reliably distinguish between 'natural fluctuations' and 'significant changes'. The longitudinal self-assessment study with repeated assessments at 1-day intervals over 14 days demonstrated the feasibility and efficiency of the speech analysis method in home environments, thus clearing the way to a broader range of applications in psychiatry.

Evaluation of mycotoxins and their metabolites in human breast milk using liquid chromatography coupled to high resolution mass spectrometry.

Humans can be exposed to mycotoxins through the food chain. Mycotoxins are mainly found as contaminants in food and could be subsequently excreted via biological fluids such as urine or human breast milk in native or metabolised form. Since breast milk is usually supposed as the only food for new-borns, the occurrence of mycotoxins in thirty-five human milk samples was evaluated by a newly developed method based on QuEChERS extraction and UHPLC-HRMS detection. The method described here allows the detection of target mycotoxins in order to determine the quality of this initial feeding. The method has been fully validated, with recoveries ranging from 64% to 93% and relative standard deviations (RSD, %) being lower than 20%. Using the method described, non-metabolised mycotoxins such as ZEA, NEO, NIV, ENA, ENA1, ENB, ENB1 and metabolites, such as ZEA metabolites, HT-2, DOM and T-2 triol were detected in human milk samples. Results obtained help to estimate the exposure of mothers and infants to mycotoxins. Moreover, to the best of our knowledge, this is the first work describing the simultaneous detection, quantification and screening of mycotoxins and their metabolites in human mature milk.

Occurrence of fumonisins in organic and conventional cereal-based products commercialized in France, Germany and Spain.

A fast and effective method for quantitative determination of fumonisins B1 and B2 (FB1 and FB2) in different cereal-based products was developed. Fumonisins were extracted combining a solid liquid extraction (SLE) pre-treatment and immunoaffinity columns (IACs), which were applied for an effective clean-up. Consecutively, high pressure liquid chromatography (HPLC) coupled to hybrid triple quadrupole-linear ion trap mass spectrometer (QTrap®) was used for the separation, detection and quantification of targeted mycotoxins. The analytical method showed acceptable recoveries ranged from 89.7% to 99.1%, as well as a relative standard deviation (%, RSD) lower than 12%. A total of 1250 samples from France, Germany and Spain were analysed. The incidence of fumonisin B1 and B2 in organic samples were 11.4% and 11.3%, respectively. By contrast, the occurrence of conventional samples was 3.6% of FB1 and 3.5% of FB2. The highest values were obtained in Spanish samples of flour, as an example toasted corn, called gofio that was contaminated with 1201.7 and 1010.5 µg kg(-1) of FB1 and FB2, respectively.