Metabolomic and transcriptomic profiles after immune stimulation in the zebrafish testes.

Fish farms are prone to disease outbreaks and stress due to high-density rearing conditions in tanks and sea cages, adversely affecting growth, reproduction, and metabolism. To understand the molecular mechanisms affected in the gonads of breeder fish after an immune challenge, we investigated the metabolome and the transcriptome profiles in zebrafish testes after inducing an immune response. After 48 h of the immune challenge, ultra-high-performance liquid chromatography (LC-MS) and transcriptomic analysis by RNA-seq (Illumina) resulted in 20 different released metabolites and 80 differentially expressed genes. Among these, glutamine and succinic acid were the most abundant metabolites released and 27,5% of the genes belong to either the immune or reproduction systems. Pathway analysis based on metabolomic and transcriptomic crosstalk identified cad and iars genes that act simultaneously with succinate metabolite. This study deciphers interactions between reproduction and immune systems and provides a basis to improve protocols in generating more resistant broodstock.

Impact of Treatment with Unilateral Implant- Supported Fixed Partial Prosthesis on the Frequency of Masticatory Side Switches in Patients with Unilateral Posterior Missing Teeth.

This study aimed to determine the impact of treatment with an implant-supported fixed partial prosthesis (ISFPP) on the frequency of masticatory side switches in patients with unilateral posterior missing teeth. This was a prospective study of 30 patients with unilateral posterior missing teeth treated with one-, two-, or three-unit ISFPPs. Comparison was with 10 healthy individuals with complete natural dentitions. Each participant performed masticatory assays, which involved chewing pieces of silicon inside a latex bag, at baseline and at 3-months' follow-up. The frequency of masticatory side switches was reported as the masticatory side-switch index: the number of side switches divided by the maximum number of possible switches. Data were analyzed by Kruskal-Wallis, Mann-Whitney U test, or Wilcoxon test, as appropriate. At baseline, the masticatory side-switch index was lower only in patients with three missing teeth than in controls. At 3 months after treatment, a significant increase in the masticatory side-switch index was only observed in patients treated with three restorative units. Treatment with ISFPPs does not change the masticatory side-switch frequency in patients with unilateral posterior missing teeth, though it could increase it in patients with three missing teeth.

Epigenetic differences in the innate response after immune stimulation during zebrafish sex differentiation.

Infections are able to trigger epigenetic modifications; however, epigenetic-mediating infections in the immune system in fish is currently unavailable. Within this purpose, zebrafish were immune-stimulated with three lipopolysaccharides (LPS) during sex differentiation. Methylation patterns of three immune genes were studied by a candidate gene approach together with gene expression analysis, and in adulthood, sex ratios were determined. It was shown that the entrance of LPS was through the gills and accumulated in the pronephros. Significant hypomethylation levels of CASP9 and a significant CpG site for IL1ß after Pseudomonas aeruginosa LPS exposure were found. No methylation difference was observed for TNFα. Gene expression and correlation data differed among studied genes. Sex ratios showed a feminization in dose and LPS strain-dependent manner. Here, it is provided epigenetic regulatory mechanisms derived by innate response and the first evidence of possible epigenetic interactions between the immune and reproductive systems.

Immune genes, IL1ß and Casp9, show sexual dimorphic methylation patterns in zebrafish gonads.

There is crosstalk between the immune and reproductive systems in which sexual dimorphism is a common pattern in vertebrates. In recent years, epigenetics has emerged as a way to study the molecular mechanisms involved in gonadal development, those responsible for integrating environmental information that contribute to assigning a specific sexual phenotype (either an ovary or a testis). The knowledge of epigenetic mechanisms in certain molecular processes allows the development of epigenetic markers. In fish gonads, the existence of reproduction-immune system interactions is known, although the epigenetic mechanisms involved are far from clear. Here, we used the zebrafish (Danio rerio) as a model to study the DNA methylation patterns in gonads of two well-known innate immune genes: IL1ß and Casp9. DNA methylation levels were studied by a candidate gene approach at single nucleotide resolution and gene expression analyses were also carried out. Results showed that there was clear sexual dimorphism in the DNA methylation levels of the two immune genes studied, being significantly higher in the testes when compared to the ovaries. In summary, and although further research is needed, this paper presents sexual dimorphic methylation patterns of two immune-related genes, thus sex-biased differences in methylation profiles should considered when analyzing immune responses in fish. Data showed here can help to develop epimarkers with forthcoming applications in livestock and fish farming production, for example, in immune fish diseases or sexual control programs as epigenetic molecular tools to predict environmental pressure in the gonads.

Characterization of the European Sea Bass (Dicentrarchus labrax) Gonadal Transcriptome During Sexual Development.

The European sea bass is one of the most important cultured fish in Europe and has a marked sexual growth dimorphism in favor of females. It is a gonochoristic species with polygenic sex determination, where a combination between still undifferentiated genetic factors and environmental temperature determines sex ratios. The molecular mechanisms responsible for gonadal sex differentiation are still unknown. Here, we sampled fish during the gonadal developmental period (110 to 350 days post fertilization, dpf), and performed a comprehensive transcriptomic study by using a species-specific microarray. This analysis uncovered sex-specific gonadal transcriptomic profiles at each stage of development, identifying larger number of differentially expressed genes in ovaries when compared to testis. The expression patterns of 54 reproduction-related genes were analyzed. We found that hsd17ß10 is a reliable marker of early ovarian differentiation. Further, three genes, pdgfb, snx1, and nfy, not previously related to fish sex differentiation, were tightly associated with testis development in the sea bass. Regarding signaling pathways, lysine degradation, bladder cancer, and NOD-like receptor signaling were enriched for ovarian development while eight pathways including basal transcription factors and steroid biosynthesis were enriched for testis development. Analysis of the transcription factor abundance showed an earlier increase in females than in males. Our results show that, although many players in the sex differentiation pathways are conserved among species, there are peculiarities in gene expression worth exploring. The genes identified in this study illustrate the diversity of players involved in fish sex differentiation and can become potential biomarkers for the management of sex ratios in the European sea bass and perhaps other cultured species.

"Cell ELISA" como ferramenta auxiliar no controle da adenite equina / "Cell ELISA" as an auxiliary tool for the control of equine adenitis

Este trabalho relata o desenvolvimento e a avaliação de um ensaio imunoenzimático (ELISA) como ferramenta auxiliar no controle da adenite equina. Foi avaliada a presença de anticorpos anti-Streptococcus equi subsp. equi em equinos com doença clínica de garrotilho, portadores assintomáticos e potros vacinados. Equinos doentes demonstraram absorbâncias médias superiores (P<0,05) às médias observadas nas demais categorias examinadas. Equinos portadores assintomáticos apresentaram valores médios de absorbância superiores (P<0,05) aos animais com cultura negativa. Logo após a vacinação, potros apresentaram elevação nos níveis de anticorpos, seguida de um decréscimo nos níveis 90 dias após a segunda vacinação. O "Cell ELISA" foi eficiente para a detecção de anticorpos em equinos expostos a antígenos de S. equi, diferenciando-se de infecções por S. zooepidemicus. O "Cell ELISA" mostrou-se uma alternativa clínica para o diagnóstico indireto da adenite equina, diferenciando-se, entre equinos assintomáticos, os potenciais portadores da infecção. Os resultados observados em potros vacinados confirmam o potencial de utilização desse teste como ferramenta em programas de vacinação contra garrotilho pelo monitoramento de rebanhos pós-vacinação. Esses resultados sugerem que o "Cell ELISA" é uma promissora ferramenta auxiliar no controle da adenite equina.(AU)

This study reports the development and evaluation of the use of "Cell ELISA" as a tool for clinical interpretation for the control of strangles. The presence of anti-S. equi antibodies was evaluated in horses with strangles, in asymptomatic carriers and in vaccinated foals. Equine positive for strangle showed higher average of absorbance (P<0.05) when compared with the average for the other categories of horses studied. Asymptomatic S. equi equine carriers had higher average of absorbance (P<0.05) than equines with negative culture. After vaccination, foals presented an increase in antibody levels, followed by a decrease in antibody levels 90 days post the second vaccination. The "Cell ELISA" was efficient for the detection of antibodies in horses exposed to S. equi antigens, differentiating infections with S. zooepidemicus. Thus, the test might be a clinical tool for indirect diagnosis of the strangles, differentiating, between the asymptomatic horses, the potential carriers of infection. The results observed in vaccinated foals confirm the potential use of this test as an auxiliary instrument for strangles vaccination programs based in the serological monitoring of the herd after immunization. These results suggest that the "Cell ELISA" is a promising auxiliary tool in the control of equine adenitis.(AU)

Differential responses to environmental challenge by common carp Cyprinus carpio highlight the importance of coping style in integrative physiology.

Common carp Cyprinus carpio displaying proactive or reactive stress coping styles were acclimated to two environmental regimes (low oxygen and low temperature), and selected groups were tested for response to an inflammatory challenge (Escherichia coli lipopolysaccharide, LPS). Plasma glucose and lactate levels were measured, as were selected C. carpio-specific messenger (m)RNA transcript abundance, including cortisol receptor (CR), enolase (ENO), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and interleukin-1-beta (IL1ß) was measured in individual whole brain samples. Basal levels (in sham injected fish held in normoxic conditions at 25° C) of plasma lactate and glucose differed between coping styles, being significantly lower in proactive individuals. Both variables increased in response to LPS challenge, with the exception of plasma glucose in reactive fish held in hypoxia. Baseline levels of gene expression under control conditions were significantly different for GAPDH between behavioural phenotypes. The responses to experimental challenge were sometimes diametrically opposed between stress-coping styles in a transcript-specific manner. For CR and GAPDH, for example, the response to LPS injection in hypoxia were opposite between proactive and reactive animals. Proactive fish showed decreased CR and increased GAPDH, whereas reactive showed the opposite response. These results further highlight that screening for stress-coping styles prior to experiments in adaptive physiology can significantly affect the interpretation of data obtained. Further, this leads to a more finely tuned analytical output providing an improved understanding of variation in individual responses to both environmental and inflammatory challenge.

Comprehensive transcriptomic analysis of the process of gonadal sex differentiation in the turbot (Scophthalmus maximus).

The turbot is a flatfish with a ZW/ZZ sex determination system but with a still unknown sex determining gene(s), and with a marked sexual growth dimorphism in favor of females. To better understand sexual development in turbot we sampled young turbot encompassing the whole process of gonadal differentiation and conducted a comprehensive transcriptomic study on its sex differentiation using a validated custom oligomicroarray. Also, the expression profiles of 18 canonical reproduction-related genes were studied along gonad development. The expression levels of gonadal aromatase cyp19a1a alone at three months of age allowed the accurate and early identification of sex before the first signs of histological differentiation. A total of 56 differentially expressed genes (DEG) that had not previously been related to sex differentiation in fish were identified within the first three months of age, of which 44 were associated with ovarian differentiation (e.g., cd98, gpd1 and cry2), and 12 with testicular differentiation (e.g., ace, capn8 and nxph1). To identify putative sex determining genes, ∼4.000 DEG in juvenile gonads were mapped and their positions compared with that of previously identified sex- and growth-related quantitative trait loci (QTL). Although no genes mapped to the previously identified sex-related QTLs, two genes (foxl2 and 17ßhsd) of the canonical reproduction-related genes mapped to growth-QTLs in linkage group (LG) 15 and LG6, respectively, suggesting that these genes are related to the growth dimorphism in this species.

Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA / Clonagem, expressão e caracterização de proteína SeM de Streptococcus equi subsp. equi e avaliação de sua utilização como antígeno em um ELISA indireto

Strangles is an economically important horse disease caused by Streptococcus equi subsp. equi. The diagnosis can be confirmed either directly by bacterial isolation and PCR or by ELISA, which is an indirect method based on the detection of serum antibodies. The aim of this study was to clone, express and characterize the SeM protein of Streptococcus equi subsp. equi, evaluate its use as antigen in indirect ELISA and determine its performance to distinguish sera of negative, vaccinated and positive animals. This was initially performed by cloning the gene encoding the SeM protein and its expression in Escherichia coli. Subsequently, the protein produced was characterized and used as antigen in ELISA. Serum samples for evaluation were taken from 40 negative foals, 46 horses vaccinated with a commercial vaccine against strangles and 46 horses diagnosed with the disease. The test showed high specificity and sensitivity, allowing discrimination between negative and positive, positive and vaccinated animals, and vaccinated animals and negative sera. Thus, it was concluded that the protein produced rSeM, which can be used as antigen for disease diagnosis, and the described ELISA might be helpful to evaluate the immune status of the herd...

A adenite equina é uma enfermidade economicamente importante de equinos, causada por Streptococcus equi subsp. equi. Seu diagnóstico pode ser confirmado de forma direta, por meio de isolamento bacteriano e de PCR, ou de forma indireta, por meio de ELISA, método baseado na detecção de anticorpos séricos. O objetivo deste estudo foi clonar, expressar e caracterizar a proteína SeM de Streptococcus equi subsp. equi, avaliar sua utilização como antígeno em um ELISA indireto e determinar a capacidade do teste de distinguir soros de animais negativos, vacinados e positivos. Para tal, foi inicialmente realizada a clonagem do gene que codifica para a proteína SeM e sua expressão em Escherichia coli. Posteriormente, a proteína produzida foi caracterizada e utilizada como antígeno em um teste de ELISA indireto. Para avaliação do teste, foram utilizadas amostras de soro de 40 potros negativos, de 46 equinos vacinados com uma vacina comercial contra adenite equina e de 46 equinos com diagnóstico da doença. O teste demonstrou alta sensibilidade e especificidade, permitindo discriminar entre soros negativos e positivos, positivos e de animais vacinados, e negativos e de animais vacinados. Assim, conclui-se que a proteína rSeM produzida pode ser usada como antígeno para o diagnóstico da enfermidade e que o ELISA descrito pode ser útil para avaliar o estado imunológico do rebanho...

Osteodistrofia fibrosa em equinos criados em pastagem de Panicum maximum cultivar Aruana: relato de casos / Fibrous osteodystrophy in horses raised on Aruana (Panicum maximum) pasture: case reports

Relata-se a ocorrência de osteodistrofia fibrosa em 38 equinos criados em pastagens de Panicum maximum cultivar Aruana, da raça Mangalarga Marchador, provenientes de uma propriedade localizada no estado do Paraná. No exame clínico geral, observou-se aumento bilateral e simétrico dos ossos da face e, também, aumento de volume na porção distal do rádio. A análise radiológica das áreas afetadas demonstrou redução da densidade óssea, e, na bioquímica sanguínea, 24 (63 por cento) animais apresentaram níveis de fósforo elevados. O exame bromatológico revelou níveis elevados de oxalato na pastagem de Aruana. Conclui-se que a ingestão de Aruana, forragem com altos níveis de oxalato, foi responsável pelo aparecimento de lesões de osteodistrofia fibrosa em equinos.

Fibrous osteodystrophy occurrence is reported in 38 Mangalarga Marchador horses raised on Aruana (Panicum maximum) pasture from a breeding farm in Paraná state. Clinical examination showed bilateral and symmetric increase in bones of face and an increase of volume at the radius distal portion. The radiologic analysis revealed bone density reduction in affected areas. Blood biochemist demonstrated high phosphorus levels in 24 animals (63 percent). Analysis of the pasture was performed and high levels of oxalate were found in the Aruana samples. It was concluded that the ingestion of Aruana, a pasture with high oxalate levels, was responsible for the appearing of fibrous osteodystrophy lesions in horses.

Use of spent mushroom substrates from Agaricus subrufescens (syn. A. blazei, A. brasiliensis) and Lentinula edodes productions in the enrichment of a soil-based potting media for lettuce (Lactuca sativa) cultivation: Growth promotion and soil bioremediation.

This study aimed to assess physicochemical and microbiological properties of fresh spent mushroom substrates (SMSs)--without post-crop heat treatment--from Agaricus subrufescens and Lentinula edodes production to optimize the use of these residues in the soil enrichment for lettuce growth promotion and soil remediation. Organic matter and C content of both SMSs were high. Fresh A. subrufescens SMS was a good source of N, P and K. On the other hand, L. edodes SMS presented a lower concentration of these nutrients and a high level of immaturity. Both SMSs presented high electric conductivity values (2.5-3.4 mS/cm). Microbiological analysis, based upon enumeration of culturable bacteria (thermophilic and mesophilic) and fungi, and also evolution of CO(2), showed that SMSs played higher microbial diversity than soil control. Laccase activity from A. subrufescens SMS tended to remain constant during a 2-month period, while L. edodes SMS presented low laccase activity throughout the same period. Agaricus subrufescens and L. edodes were able to grow on a PDA (Potato Dextrose Agar) media supplemented with different concentrations of atrazine (1-50 microg/ml), degraded the herbicide, attaining rates of 35% and 26%, respectively. On experiments of lettuce growth promotion using a soil-based potting media with different SMS rates, 5% and 10% (dw) rates of A. subrufescens SMS resulted in higher lettuce aerial dry weights than the rates of 25% and 40%, the chemical fertilization (NPK) and the control (soil). At 10% supplementation, lettuce aerial dry weight increased 2.2 and 1.3 times compared to the control and the NPK treatment, respectively. Protein content increased along with SMS rates. Fresh A. subrufescens SMS was an excellent supplement for lettuce growth promotion and showed potential for remediation of biocides possibly due to improved microbial diversity and enzymatic activity. Fresh L. edodes SMS was not a good fertilizer, at least under the conditions tested. However, microbiological analysis showed that promising results may be achieved when using fresh L. edodes SMS for soil remediation.

CD83 expression in sea bream macrophages is a marker for the LPS-induced inflammatory response.

CD83, a cell surface membrane glycoprotein member of the Ig superfamily which is commonly used as standard surface marker for dendritic cells, was cloned from gilthead sea bream macrophages using degenerate primers against conserved motifs of known CD83 sequences. The obtained cDNA contains an open reading frame of 669 nucleotides that translate into a 222 amino acid putative peptide. The deduced protein sequence shows conservation of features shared by vertebrate CD83 and multiple alignment with fish CD83 sequences reveals high homology. In cultured sea bream macrophages CD83 mRNA expression was significantly enhanced in a dose- and time-dependent fashion after stimulation with Escherichia coli LPS. These results indicate that in fish, macrophages express high levels of CD83 mRNA after LPS exposure and CD83 is therefore a good marker for activated mature myeloid cells in fish.

Increase in prevalence of rhinoconjunctivitis but not asthma and atopic eczema in teenagers.

OBJECTIVE: We sought to assess changes in the prevalence of asthma and other allergic diseases in schoolchildren of Curitiba between 1995 and 2001. METHODS: Schoolchildren (aged 13-14) were evaluated by a self-reported written questionnaire of ISAAC protocol for symptoms of asthma and allergic diseases. The diagnoses of asthma, rhinoconjunctivitis and atopic eczema were considered according to the ISAAC protocol. Probable asthma was considered: more than 4 attacks of wheezing in the last 12 months; 1 to 3 attacks of wheezing in the last 12 months with dry cough at night, and/or with sleep disturbance due to wheezing, and/or with wheezing after exercise. The data obtained in 2001 were compared with the data obtained in 1995 for the same age group. RESULTS: We evaluated 2,946 schoolchildren in 1995 and 3.628 in 2001. The prevalence of symptoms of asthma, probable asthma, rhinoconjunctivitis and eczema in 1995 and 2001 was 18.4% and 18.7%, 11.6% and 12.4%, 14.2% and 17.2%, 3.8% and 3.7%, respectively. There was a 23.7% increase in the prevalence of rhinoconjunctivitis symptoms (p < 0.05). CONCLUSIONS: The prevalence of symptoms of asthma, probable asthma and eczema did not change over the past 6 years but a significant increase of the rhinoconjunctivitis symptoms was detected.

Olive oil and the Mediterranean diet: beyond the rhetoric.

OBJECTIVE: The purpose of this study was to analyze the association of food, nutrient and energy intakes with olive oil consumption in Spain. DESIGN: Cross-sectional study by face-to-face interview. SETTING: Population-based random sample derived from the Catalan Nutrition Survey. SUBJECTS: In total, 1600 individuals between 18 and 60 y of age. INTERVENTION: Two 24-h recalls were administered to measure food and nutrient intakes. Food consumption and nutrient intakes were analyzed comparing the highest and lowest quartiles of olive oil consumption. RESULTS: Those with the highest consumption of olive oil (greater than 13.5% of total calories, fourth quartile) consumed less cereal, baked goods, whole milk, sausages, candy, fruit juice and soft drinks, but more fish, eggs, vegetables and added fats, as compared to those with the lowest olive oil consumption (less than 6.8% of total calories, first quartile). The group with the greatest olive oil consumption also demonstrated high total fat intake, although saturated fats showed a lower percentage of total energy intake. Vitamin intake was more adequate in those with the highest consumption of olive oil. CONCLUSION: Olive oil is a key contributor to the healthy aspects attributed to the Mediterranean diet, and as such, nutritional objectives in Mediterranean countries should address reducing saturated fats, without modifying quantities of olive oil.

Nutrient adequacy and Mediterranean Diet in Spanish school children and adolescents.

OBJECTIVE: To evaluate dietary habits and nutritional status of Spanish school children and adolescents, and their relationship with the Mediterranean Diet. DESIGN: Cross-sectional study by face-to-face interview. SETTING: Free living children and adolescents of all Spanish regions. SUBJECTS: A random sample of 3166 people aged 6-24 y. METHODS: Home interviews conducted by a team of 43 dietitians included 24-h recalls (a second 24-h recall in 25% of the sample) and a short frequency questionnaire to determine the quality of the Mediterranean Diet (KIDMED). RESULTS: The participation rate was 68%. In general, the adequacy of the Mediterranean Diet rose with increased mean intakes of the majority of vitamins and minerals and decreased percentages of inadequate intakes (&<2/3 of the RNI) for calcium, magnesium, vitamin B(6) and C in both sexes, and iron and vitamin A only in females. CONCLUSIONS: This study demonstrates that the Mediterranean Diet contributes to nutritional quality, and also shows concomitant risks as the Mediterranean Diet deteriorates.

Sociodemographic and lifestyle determinants of food patterns in Spanish children and adolescents: the enKid study.

OBJECTIVE: To analyse prevailing food patterns among Spanish children and young people and their relationship to sociodemographic and lifestyle factors. DESIGN: Cross-sectional population survey. SETTING: Population study. Data were collected at participants' home addresses. SUBJECTS: Random sample of the Spanish population aged 2-24 y (n=3534; 1629 boys and 1905 girls). INTERVENTIONS: Food consumption was assessed by means of a 24-h recall and a food frequency questionnaire. Sociodemographic and lifestyle related data were collected by specially designed questionnaires, previously pretested and validated. All the information was collected during a personal interview by trained dietitians. DATA COLLECTION: May 1998-April 2000. RESULTS: Average consumption of fruit and vegetables was low. The youngest age group (2-5 y) showed the lowest proportions of inadequacy for the dairy group (P<0,001; chi(2)=39.11 boys; chi(2)=49.60 girls). Factor analysis identified five main components of dietary patterns. The 'Snacky' pattern was characterised by more frequent and higher consumption of bakery products (buns, cakes and biscuits), sweets, salted snacks and soft drinks. Higher intakes of fruit, vegetables and fish were associated to the 'Healthy' pattern. Children whose mother had a low level of education and those who spent more than 2 h daily watching TV were more likely to follow the 'Snacky' pattern. Girls were more likely to follow the 'Healthy' pattern, while children and young people whose mother had a lower level of education were less likely. CONCLUSION: Results from this study highlight the importance of enhancing school-based and community-based actions to promote healthy eating and physical activity addressed to children and young people.

Food preferences of Spanish children and young people: the enKid study.

OBJECTIVE: To describe the profile of food preferences, likes and dislikes of Spanish children and young people and analyse its connection with prevailing food patterns. DESIGN: Cross-sectional population survey. SETTING: Population study. Data were collected at participants' home address. SUBJECTS: Random sample of the Spanish population aged 2-24 y (n=3534; 1629 boys and 1905 girls). INTERVENTIONS: Food preferences, food consumption and practices as well as nutrition-related information were assessed by means of a multiple choice questionnaire. A food preference score was computed considering food items ranked as first, second or third choice within each food group. Data collection was carried out during May 1998-April 2000. RESULTS: Bananas and apples were the fruit items preferred by Spanish children and young people across all age and gender groups. Within the vegetable group tomato sauce and salads, particularly lettuce and tomato salad scored highest, followed by carrots in all age and gender groups. However, 47% (95% confidence interval 46-48%) of the sample reported dislike for vegetables and an additional 5.7% (95% confidence interval 4.9-6.5%) a dislike for fruit. The proportion of individuals with low consumption of vegetables or fruit was significantly higher among those reporting a dislike either for vegetables (chi(2)=127.69; P<0.001); fruit (chi(2)=24.62; P<0.001) or for both groups (chi(2)=81.53; P<0.001). CONCLUSION: There is a significant relationship between the likes/dislikes for fruits and vegetables and usual consumption of this food group among children and young people. Strategies addressed to improve acceptance for this food group should be considered when designing interventions aimed at promoting adequate consumption of fruits and vegetables among children and young people.

A xyloglucan from seeds of the native Brazilian species Hymenaea courbaril for micropropagation of Marubakaido and Jonagored apples.

Xyloglucan was extracted from seeds of Hymenaea courbaril and mixed with agar to prepare a solid culture medium used for micropropagation of the Marubakaido apple rootstock (Malus prunifolia Borkh) and cv. Jonagored (Malus domestica). The performance on gels created from a blend of 0.4%agar and 0.2% xyloglucan (w/v) was compared with that on media gelled with a standard concentration 0.6% (w/v) of agar. The growth of shoots and the multiplication rate were higher on the modified culture medium than on the agar-gelled medium. The occurrence of hyperhydric shoots was lower on the modified medium. In the absence of auxin, shoot rooting reached 70% (Marubakaido) and 66% (Jonagored) on the agar-xyloglucan medium and 6.7% and 10.4%, respectively, on the agar medium. When 0.25 microM indole-3-butyric acid (IBA) was added to both media, the modified medium gave better results in terms of rooting percentage and quality of roots than the agar-gelled medium.

Comparative analysis of nutrition data from national, household, and individual levels: results from a WHO-CINDI collaborative project in Canada, Finland, Poland, and Spain.

STUDY OBJECTIVE: This project determined to what extent data on diet and nutrition, which were collected in a non-uniform manner, could be harmonised and pooled for international and national comparison. DESIGN: Direct comparisons of dietary data between studies were made using food balance sheets (FBS), household budget surveys (HBS), and individual dietary data (IDS); comparisons were also made within countries. Differences in study design and methodological approaches were taken into consideration. Data from research projects from the following four World Health Organisation (WHO) Countrywide Integrated Noncommunicable Disease Intervention (CINDI) countries were included-Canada, Finland, Poland, and Spain. MAIN RESULTS: FBS overestimated food consumption and nutrient intake compared to IDS. Results between HBS and IDS were quite similar, except for fish, meat, pulses and vegetables, which were underestimated by HBS, and sugar and honey and cereals, which were overestimated. Percentages of energy from fat, carbohydrates and proteins were higher when estimated from FBS, HBS, and IDS respectively. CONCLUSIONS: Results suggest that estimations from these three sources of dietary data are difficult to compare because they are measuring different levels of dietary information. The understanding of their relations may be important in formulating and evaluating a nutrition policy.

Mediterranean diet and health: is all the secret in olive oil?

OBJECTIVE: The purpose of this study was to review and analyze the synergies of olive oil related food consumption and nutrient intake in a Mediterranean region. DESIGN: Cross sectional study by face to face interview. SETTING: Population based random sample derived from the Catalan Nutrition Survey. SUBJECTS: 1600 individuals between 18 and 60 years of age. INTERVENTION: Two 24-hour recalls were administered to measure food and nutrient intakes. Food group consumption and nutrient intakes ratios between the highest(greater than 13.5% of total calories, 4th quartile)and lowest quartiles (less than 6.8% of total calories,1st quartile) of olive oil consumption were calculated. RESULTS: Comparing the highest quartile of olive oil consumption to the lowest, the highest ratios by food group consumption were observed for eggs, vegetables and fish in both men and women and non fried potatoes in women only. Lower ratios were associated in both sexes with soft drinks, commercially baked goods, processed meat and full fat milk and nuts in women only, and fried potatoes in men only. Analysis by nutrients showed higher ratios for carotenoids,monounsaturated fatty acids, vitamin E and folic acid in both sexes and Vitamin C and polyunsaturated fatty acids in women only. Lower ratios were seen for mono,di and polysaccharides, saturated fat, retinol and sodium in both genders and calcium niacin, thiamin,riboflavin and fibre in men only. CONCLUSION: Olive oil is a key contributor to the healthy aspects attributed to the Mediterranean diet, since its own health attributes may be added to those of foods that it's commonly served with (fish, vegetables), thus contributing to its thrombosis related disease preventive properties. As such, nutritional objectives in Mediterranean countries should address reducing saturated fats, without modifying quantities of olive oil,and increasing fish, vegetables and nuts.