Clonación y expresión de proteínas recombinantes del virus de la hepatitis E en sistemas heterólogos: caracterización y aplicaciones

Tesis doctoral inédita. Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 16-07-2010Bibliografí

X-Ray Structure and Molecular-Packing Analysis of Artemetin

5-Hydroxy-3,3' ,4',6,7-pentamethoxyflavone (flavone is 2-phenyl-4H-1-benzopyran-4-one ), C20H20- 08, M,= 388·4, monoclinic, P2/n, a= 7·518 (4), b=13·625(3), C=17·766(5)Á, P=98·52(3) 0 , V = 1803 (l)Á\ Z=4, Dx= 1·430Mgm-J, A.(MoKa) = 0·7107 A, !J = 0·104 mm-I, F(OOO) = 816, room temperature, final wR = 0·063 for 1480 observed reflexions. The benzopyran ring and the attached phenyl ring are quasi-planar. Dihedral angles between least-squares planes through each of the two rings are lower than 2°. An intramolecular 0-H ···O hydrogen bond exists involving hydroxyl and carbonyl groups of the phenyl and pyrone rings. Molecular-packing analysis in the atom-atom approach yields an equilib­ rium configuration in very good agreement with the experimental one

Manganese Stress Tolerance Depends on Yap1 and Stress-Activated MAP Kinases

Understanding which intracellular signaling pathways are activated by manganese stress is crucial to decipher how metal overload compromise cellular integrity. Here, we unveil a role for oxidative and cell wall stress signaling in the response to manganese stress in yeast. We find that the oxidative stress transcription factor Yap1 protects cells against manganese toxicity. Conversely, extracellular manganese addition causes a rapid decay in Yap1 protein levels. In addition, manganese stress activates the MAPKs Hog1 and Slt2 (Mpk1) and leads to an up-regulation of the Slt2 downstream transcription factor target Rlm1. Importantly, Yap1 and Slt2 are both required to protect cells from oxidative stress in mutants impaired in manganese detoxification. Under such circumstances, Slt2 activation is enhanced upon Yap1 depletion suggesting an interplay between different stress signaling nodes to optimize cellular stress responses and manganese tolerance.Universidad de Sevilla 2020/00001326Junta de Andalucía P20-RT-01220Ministerio de Ciencia e Innovación PID2019-105342GB-I0

West Nile Virus Replication Requires Fatty Acid Synthesis but Is Independent on Phosphatidylinositol-4-Phosphate Lipids

West Nile virus (WNV) is a neurovirulent mosquito-borne flavivirus, which main natural hosts are birds but it also infects equines and humans, among other mammals. As in the case of other plus-stranded RNA viruses, WNV replication is associated to intracellular membrane rearrangements. Based on results obtained with a variety of viruses, different cellular processes have been shown to play important roles on these membrane rearrangements for efficient viral replication. As these processes are related to lipid metabolism, fatty acid synthesis, as well as generation of a specific lipid microenvironment enriched in phosphatidylinositol-4-phosphate (PI4P), has been associated to it in other viral models. In this study, intracellular membrane rearrangements following infection with a highly neurovirulent strain of WNV were addressed by means of electron and confocal microscopy. Infection of WNV, and specifically viral RNA replication, were dependent on fatty acid synthesis, as revealed by the inhibitory effect of cerulenin and C75, two pharmacological inhibitors of fatty acid synthase, a key enzyme of this process. However, WNV infection did not induce redistribution of PI4P lipids, and PI4P did not localize at viral replication complex. Even more, WNV multiplication was not inhibited by the use of the phosphatidylinositol-4-kinase inhibitor PIK93, while infection by the enterovirus Coxsackievirus B5 was reduced. Similar features were found when infection by other flavivirus, the Usutu virus (USUV), was analyzed. These features of WNV replication could help to design specific antiviral approaches against WNV and other related flaviviruses

Widespread distribution of hepatitis E virus in Spanish pig herds

<p>Abstract</p> <p>Background</p> <p>Hepatitis E virus (HEV) infection is a serious health problem in developing countries and is also increasingly reported in industrialized regions. HEV is considered a zoonotic agent and strains isolated from swine and human sources are genetically similar. Thus, HEV is of increasing importance to both public and animal health. The aim of the present study was to evaluate the distribution of HEV in a large population of pigs from herds located in different autonomous regions throughout Spain.</p> <p>Results</p> <p>The presence of anti-HEV IgG antibodies was analyzed in 1141 swine serum samples (corresponding to 381 pigs younger than 6 months and 760 pigs older than 6 months) collected from 85 herds. Herds were located in 6 provinces in 4 autonomous regions throughout Spain. At least one pig tested positive for anti-HEV IgG in over 80% of herds. Of individual pigs, 20.4% (233/1141) were positive for anti-HEV IgG, with the prevalence being higher in adult pigs than in those under 6 months (30.2% <it>vs. </it>15.5%). A subset of serum samples taken at 2- to 5-week intervals showed that seroprevalence dropped between 3 and 11 weeks of age, and then rose significantly by the 15th week. Pigs were also examined for the presence of HEV-RNA by RT-PCR. Of pigs tested for the presence of HEV-RNA 18.8% (64/341) were positive, with at least one pig in almost half of the herds testing positive. HEV-RNA amplicons from several positive pigs were sequenced and all were of genotype 3.</p> <p>Conclusions</p> <p>HEV was found to be widely distributed among swine farms across Spain, with the prevalence being highest among animals older than 6 months. These results indicate that HEV infection either is or is likely to become endemic in the Spanish swine population.</p

Spanish children's diet: compliance with nutrient and food intake guidelines

OBJECTIVE: To compare the diet of Spanish children against the nutrient and food intake guidelines. To calculate an index of overall diet quality and check its validity against nutrient intake. DESIGN AND SETTING: Cross-sectional study in four cities in Spain, where information on food and nutrient intake was obtained from schoolchildren through a food frequency questionnaire. PARTICIPANTS: The sample included 1112 children (overall response rate of 85%) attending public and private schools and aged 6-7 y. Children were selected through random cluster sampling in schools, and stratified by sex and socioeconomic level. MAIN OUTCOME MEASURES: Mean nutrient intake, number of food servings, and the percentage of children who meet recommended nutrient and food-serving intake levels. The overall dietary quality was assessed using the Healthy Eating Index (HEI). RESULTS: Mean micronutrient intake exceeded 100% of the recommended dietary allowances, except for vitamin B6, which registered a mean intake of 77.1%. For almost all children, intake of saturated fat was above, and that of carbohydrate below, the recommended level, in contrast to the relatively high compliance with the recommendations for poly- and monounsaturated fatty acid, salt and fiber intake (69.7, 43.7, 40.7, and 30.1%, respectively). Consumption of food servings for each of the five American pyramid food groups came close to or exceeded USDA guidelines, with the exception of cereals, with 5.4 servings per day. The mean score obtained in the HEI was 64.6. Children who complied with all the food guide pyramid recommendations registered a higher dietary variety and a healthier nutritional profile. CONCLUSIONS: Children aged 6-7 y show scant compliance with the macronutrient goals for healthy eating. Micronutrient intake is adequate in general, yet there are small groups of children with risk of deficient intake of vitamins B6 and D. While Spanish children's eating habits are reasonably in line with American food guide pyramid guidelines, consumptions of cereals and fruit should be improved.This study was partly funded by grants from the Inter-national Olive Oil Board (Consejo Oleı ́cola Internacional),Comunidad de Madrid, Fundacio ́n Pedro Barrie ́de laMaza, and Fundacio ́n Eugenio Rodrı ́guez Pascua

Food sources of nutrients in the diet of Spanish children: the Four Provinces Study

The aim of the present study was to assess the principal food sources of energy and nutrients among Spanish children. We used a cross-sectional study design, based on results obtained from a food-frequency questionnaire. The sample included 1112 children, aged 6-7 years, from Cadiz, Madrid, Orense and Murcia, Spain. Children were selected through random cluster-sampling in schools. We analysed the percentage contributed by each food item to total energy and nutrient intake. The most important food sources were: white bread in the case of carbohydrate (13.4 %); olive oil in the case of total lipids (18.3 %) and monounsaturated fatty acids (29.2 %); whole milk in the case of protein (10.2 %) and saturated fatty acids (14.9 %); chips (French fried potatoes) in the case of polyunsaturated fatty acids (30.4 %). The greatest proportion of Na, consumed in excess, came from salt added to meals. Ham ranked second as a source of saturated fats. Fruits and green leafy vegetables proved to have great relevance as sources of fibre and vitamins, though with regard to the latter, it was observed that fortified foods (breakfast cereals, dairy products, fruit juices, etc.) had come to play a relevant role in many cases. In conclusion, the nutritional profile of Spanish school-aged children aged 6-7 years could be improved by nutritional policies targeted at limiting their consumption of ham (cured or cooked) and of salt added to meals, replacing whole milk with semi-skimmed milk, encouraging the consumption of products rich in complex carbohydrates already present in children's diets (bread, pasta, rice) and promoting less fatty ways of cooking food.This study was partly funded by grants from the Inter-national Olive Oil Board (Consejo Oleı ́cola Internacional),Madrid Regional Authority (Comunidad de Madrid), PedroBarrie ́de la Maza Foundation and Eugenio Rodrı ́guezPascual FoundationS

Consumption of bakery products, sweetened soft drinks and yogurt among children aged 6-7 years: association with nutrient intake and overall diet quality

The present study tests the hypothesis that higher consumption of bakery products, sweetened soft drinks and yogurt is associated with higher intake of energy, saturated fats, sugars and worse overall diet quality among Spanish children. This is a cross-sectional study covering 1112 children aged 6.0-7.0 years in four Spanish cities. Nutrient and food intake were obtained through a food-frequency questionnaire, and overall diet quality calculated using the healthy-eating index (HEI) developed by Kennedy et al. (1995). Standardized methods were used to measure anthropometric variables. Associations of interest were summarized as the difference in nutrient and food consumption between the value of the fifth and the first quintile of consumption (dq) of bakery products, sweetened soft drinks or yogurt, adjusted for energy intake and BMI. Bakery products, sweetened soft drinks and yogurt supplied 15.5, 1.0 and 5.6 % energy intake respectively. Higher consumption of these three foods was associated with greater energy intake (P<0.001), but not with higher BMI. Consumption of bakery products was associated with the proportion of energy derived from intake of total carbohydrates (dq 4.5 %, P<0.001) and sugars (dq 2 %, P<0.001), but did not show association with the HEI. Consumption of sweetened soft drinks was associated with a lower consumption of milk (dq -88 ml, P<0.001) and Ca (dq -175 mg/d, P<0.001), and worse HEI (dq -2, P<0.01). Consumption of yogurt, while associated with higher energy intake from saturated fats (dq 1.77 %, P<0.001) and sugars (dq 2.02 %, P<0.001), showed no association with the HEI. Differences in the intake of nutrients and foods across quintiles of consumption of bakery products, sweetened soft drinks and yogurt were usually very small. We conclude that the impact of the consumption of bakery products, sweetened soft drinks and yogurt on the quality of the diet of Spanish children is only modest, although it may contribute to aggravating certain unhealthy characteristics of their diet, particularly excess energy, saturated fats and sugars. Therefore, consumption of bakery products and sweetened soft drinks should be moderated, and priority given to consumption of low-fat, low-sugar yogurt.This study was partly funded by grants from the Inter-national Olive Oil Board (Consejo Oleı ́cola Internacional),Comunidad de Madrid, Fundacio ́n Pedro Barrie ́de laMaza, and Fundacio ́n Eugenio Rodrı ́guez PascualS

Greater dietary variety is associated with better biochemical nutritional status in Spanish children: the Four Provinces Study

BACKGROUND AND AIM: Although dietary variety has been associated with a better nutritional profile, its possible role in obesity raises doubts about its overall health benefits. In this study, we examined the association between dietary variety and anthropometric variables, food intake and various food intake biomarkers in Spanish children. METHODS AND RESULTS: This was a cross-sectional study of 1112 children aged 6-7 years from Cadiz, Murcia, Orense and Madrid, who were selected by means of the random cluster-sampling of schools. Information concerning food and nutrient intake was obtained using a food frequency questionnaire, and a dietary variety index (DVI) was calculated on the basis of the number of different foods consumed more than once a month. The anthropometric variables (weight and height), and plasma lipid and vitamin levels were determined using standardised methods. Our results show that the body mass index (BMI) did not vary substantially as a function of DVI: it was 16.9 in the lowest DVI tertile and 17.2 in the highest (p=0.20). Unlike BMI, the DVI positively correlated (p<0.05) with the plasma levels of alpha and beta-carotene, lycopene, retinol, alpha-tocopherol and vitamin E, with energy intake, and with most of the foods, particularly vegetables, fruit and sausages (respective correlation coefficients of 0.43, 0.26 and 0.23). CONCLUSIONS: Dietary variety is associated with a better food and nutritional profile in Spanish children. Nevertheless, the presence of a positive association between the DVI and energy intake, and the consumption of sausages and pre-cooked products calls for the recommendation of a varied diet of healthy foods, such as cereals (especially whole grains), fruits and vegetables