Aplicación de nuevas tecnologías para la detección de reordenamientos genómicos en pacientes con retraso del neurodesarrollo y anomalías congénitas

El retraso del neurodesarrollo o discapacidad intelectual (DI) es una de las causas de minusvalías más frecuente en niños, afecta a cuatro de cada mil nacidos vivos en su forma más grave, aunque el retraso leve podría afectar a 1-3% de la población. La mayoría de los casos de DI son de origen desconocido. Las variantes de número de copias (CNVs) son una causa importante de discapacidad intelectual sindrómica. A lo largo de estos años se han desarrollado diferentes herramientas para el estudio genético de la discapacidad intelectual sindrómica, tales como la FISH (hibridación in situ fluorescente), el MLPA (multiplex ligation-dependent probe amplification) o la CGH-array (Hibridación genómica comparada sobre arrays). La hipótesis es que los reordenamientos genómicos son una causa importante de retraso del neurodesarrollo y anomalías congénitas. La aplicación de diferentes tecnologías nos va a permitir evaluar qué método es el más adecuado para su detección y aplicación en la práctica clínica. El objetivo general de este trabajo es evaluar la aplicabilidad de las diferentes tecnologías (FISH, MLPA y CGH-array) para la detección de reordenamientos genómicos desequilibrados en pacientes con retraso psicomotor y anomalías congénitas. Se ha estudiado una serie de 202 pacientes con retraso del neurodesarrollo asociado a anomalías congénitas, antecedentes familiares de DI y/o abortos de repetición, mediante FISH y MLPA de regiones subteloméricas y dos tipos de arrays genómicos (basado en BACs y en oligonucleótidos). Uno de los grandes retos de este trabajo ha sido la interpretación de la repercusión clínica de las CNVs detectadas. Inicialmente se utilizaron diferentes criterios, clasificando las CNVs en patológicas si habían surgido de novo o presentaban un tamaño mayor de 1Mb y polimórficas si habían sido heredadas de un progenitor sano o se habían descrito en población general. A lo largo de este trabajo, se ha puesto de manifiesto que no existen criterios únicos con validez universal para establecer el carácter patológico o polimórfico de las CNVs, hoy por hoy, la interpretación es muy dinámica, y está sujeta al conocimiento disponible en cada momento. La frecuencia de reordenamientos considerados patológicos detectados mediante FISH y MLPA ha sido similar, aproximadamente del 10%, siendo la técnica del MLPA la más indicada de las dos para el rastreo de reordenamientos subteloméricos, por ser más económica y menos laboriosa. Mediante la CGH-array se han detectado un mayor número de CNVs patológicas, un 12% mediante el array basado en BACs y un 25% mediante el array basado en oligonucleótidos. Por otra parte, el array de oligonucleótidos presenta una mayor especificidad, sensibilidad y posibilidad de automatización. Con estos resultados concluimos que la CGH-array basada en oligonucleótidos es la técnica más apropiada para la detección de reordenamientos genómicos desequilibrados en todo el genoma. El número de CNVs patológicas detectadas presentan una buena correlación con el tamaño del cromosoma, sugiriendo una distribución uniforme, salvo en los cromosomas X, 16 y 22 que presentan una mayor tasa de alteraciones en relación a su tamaño. Finalmente, para poder ofrecer un asesoramiento genético adecuado a las familias tras la detección de CNVs patológicas, es conveniente realizar estudios complementarios mediante la técnica del FISH para detectar reordenamientos genómicos equilibrados y translocaciones intersticiales en los progenitores.Neurodevelopmental delay or intellectual disability (ID) is one of the most common causes of disability in children (4/1000). The majority of ID cases have an unknown origin. Copy number variants (CNVs) are a cause of syndromic intellectual disability. In these last years, new tools have been developed for the genetic study of ID, such as FISH (fluorescence in situ hybridization), the MLPA (multiplex ligation -dependent probe amplification) or CGH-array (Comparative genomic hybridization arrays). The hypothesis is that genomic rearrangements are an important cause of neurodevelopmental delay and congenital anomalies. The application of different technologies will allow us to evaluate which method is best suited for detection and application in clinical practice. The objective of this work is evaluate the applicability of these technologies for detecting unbalanced genomic rearrangements in patients with syndromic ID. We have studied 202 patients with neurodevelopmental delay associated with congenital abnormalities, family history of ID and / or repeat abortions, by subtelomeric FISH and MLPA and two types of genomic arrays (based on BACs and oligonucleotides). One of the great challenges of this work was the clinical interpretation of the CNVs detected. Initially different simple criteria were used in classifying pathogenic CNVs if they had arisen de novo or had a size larger than 1Mb and polymorphic if they were inherited from a healthy parent or had been described in general population. In this work, it has been shown that there are no unique criteria with universal validity to establish the pathogenic or polymorphic character of CNVs. Today the interpretation is very dynamic and is subject to the knowledge available at the time. The pathogenic CNVs frequencies detected by FISH and MLPA were similar (10 %), but the MLPA technique was better for subtelomeric rearrangements screening, being cheaper and less laborious. By CGH-array was detected a greater number of pathogenic CNVs, 12% by BACs array and 25% by oligonucleotide array. Moreover, the oligonucleotides array was a higher specificity, sensitivity and automation possibility. With these results, we concluded that the CGH-array based on oligonucleotide is the most appropriate technique for the detection of genomic rearrangements unbalanced. A good correlation of the number of CNVs and the size of the chromosomes suggests a uniform distribution, except for chromosomes X, 16 and 22, which have a higher rate of changes in relation to its size. Finally, in order to offer appropriate genetic counseling to families following the detection of pathogenic CNVs, it is advisable to perform additional studies using FISH technique to detect genomic rearrangements and interstitial balanced translocations in parents

Enrichment of ultraconserved elements among genomic imbalances causing mental delay and congenital anomalies

<p>Abstract</p> <p>Background</p> <p>The ultraconserved elements (UCEs) are defined as stretches of at least 200 base pairs of human DNA that match identically with corresponding regions in the mouse and rat genomes, albeit their real significance remains an intriguing issue. These elements are most often located either overlapping exons in genes involved in RNA processing or in introns or nearby genes involved in the regulation of transcription and development. Interestingly, human UCEs have been reported to be strongly depleted among segmental duplications and benign copy number variants (CNVs). However no comprehensive survey of a putative enrichment of these elements among pathogenic dose variants has yet been reported.</p> <p>Results</p> <p>A survey for UCEs was performed among the 26 cryptic genomic rearrangements detected in our series of 200 patients with idiopathic neurodevelopmental disorders associated to congenital anomalies. A total of 29 elements, out of the 481 described UCEs, were contained in 13 of the 26 pathogenic gains or losses detected in our series, what represents a highly significant enrichment of ultraconserved elements. In addition, here we show that these elements are preferentially found in pathogenic deletions (enrichment ratio 3.6 vs. 0.5 in duplications), and that this association is not related with a higher content of genes. In contrast, pathogenic CNVs lacking UCEs showed almost a threefold higher content in genes.</p> <p>Conclusions</p> <p>We propose that these elements may be interpreted as hallmarks for dose-sensitive genes, particularly for those genes whose gain or loss may be directly implied in neurodevelopmental disorders. Therefore, their presence in genomic imbalances of unknown effect might be suggestive of a clinically relevant condition.</p

Impact of the Content of Fatty Acids of Oral Fat Tolerance Tests on Postprandial Triglyceridemia: Systematic Review and Meta-Analysis

Whether the content of saturated (SFA), monounsaturated (MUFA), and polyunsaturated fatty acids (PUFA) could differently influence postprandial triglycerides (TG) is unknown. We examined possible differences in the postprandial TG response to fat tolerance tests (FTTs), in which SFA or unsaturated fatty acids were used. Crossover clinical trials investigating the effects of FTTs containing SFA and unsaturated fats on postprandial triglyceridemia in databases from 1994 until 2016 were searched. Of 356 studies, 338 were excluded and 18 were considered. TG net incremental areas under the curve were calculated using time-points or changes from baseline. Pooled effects of standardized mean differences and I2 test were used. Results: In 12 studies, responses to SFA versus PUFA meals, and in 16 studies versus MUFA meals were compared. Over 4 hours, no differences between SFA and unsaturated fats were observed. Over 8 hours a lower response to PUFA (SMD −2.28; 95%CI −4.16, −0.41) and a trend to lower response to MUFA (SMD −0.89, 95%CI −1.82, 0.04) were detected. FTTs shorter than 8 hours may not be sufficient to differentiate postprandial TG after challenges with distinct fatty acids. Clinical significance of different postprandial TG responses on cardiovascular risk in the long-term deserves investigation

Distinct breakfast patterns on satiety perception in individuals with weight excess

ABSTRACT Objectives: Western dietary pattern predisposes to weight gain, insulin resistance and cardiometabolic diseases. Promoting satiety via modifications in diet composition could be useful to fight weight gain. Mediterranean diet which is recognized to be cardioprotective contains high fiber and unsaturated fat contents. We compared the effects of distinct breakfast patterns on satiety of individuals at cardiometabolic risk, and examined the correlation of satiety level after each breakfast intervention period with glucose parameters. Materials and methods: In this 10-week cross-over clinical trial, 54 individuals with weight excess were submitted to 2 types of 4-week isocaloric breakfasts (2-week washout), one typically Brazilian and a modified one, differing concerning fiber and types of fatty acids contents. Clinical data were collected before and after each breakfast. A satiety scale was applied at fasting and 10, 30 and 120&apos; after breakfast consumption. Repeated measures ANOVA, Student t test or non-parametric correspondents were used; correlations were tested by Pearson or Spearman coefficients. Results: Anthropometric variations after breakfasts were not significant. Only after the modified breakfast, reduction in blood pressure levels was observed. The satiety level did not show significant variation across each period or between the breakfasts. Non-significant correlation between satiety and glucose, insulin and HOMA-IR values after each intervention period was observed. Conclusion: We conclude that different breakfast compositions do not alter satiety level, which is not correlated to glucose parameters in overweight individuals. Stronger modifications of daily meals might be necessary to differentiate satiety levels under distinct dietary patterns. Arch Endocrinol Metab. 2016;60(4):333-4

Distinct breakfast patterns on satiety perception in individuals with weight excess

ABSTRACT Objectives: Western dietary pattern predisposes to weight gain, insulin resistance and cardiometabolic diseases. Promoting satiety via modifications in diet composition could be useful to fight weight gain. Mediterranean diet which is recognized to be cardioprotective contains high fiber and unsaturated fat contents. We compared the effects of distinct breakfast patterns on satiety of individuals at cardiometabolic risk, and examined the correlation of satiety level after each breakfast intervention period with glucose parameters. Materials and methods: In this 10-week cross-over clinical trial, 54 individuals with weight excess were submitted to 2 types of 4-week isocaloric breakfasts (2-week washout), one typically Brazilian and a modified one, differing concerning fiber and types of fatty acids contents. Clinical data were collected before and after each breakfast. A satiety scale was applied at fasting and 10, 30 and 120&apos; after breakfast consumption. Repeated measures ANOVA, Student t test or non-parametric correspondents were used; correlations were tested by Pearson or Spearman coefficients. Results: Anthropometric variations after breakfasts were not significant. Only after the modified breakfast, reduction in blood pressure levels was observed. The satiety level did not show significant variation across each period or between the breakfasts. Non-significant correlation between satiety and glucose, insulin and HOMA-IR values after each intervention period was observed. Conclusion: We conclude that different breakfast compositions do not alter satiety level, which is not correlated to glucose parameters in overweight individuals. Stronger modifications of daily meals might be necessary to differentiate satiety levels under distinct dietary patterns

Survey of the occurrence of Giardia duodenalis cysts and Cryptosporidium spp. oocysts in green leafy vegetables marketed in the city of Valencia (Spain)

The role of vegetables usually consumed without prior culinary treatment is known to contribute to the prevalence of foodborne diseases. Cysts and oocysts can contaminate food, which can then be the source of infection in humans. The aim of the study was to assess the occurrence of Giardia duodenalis and Cryptosporidium spp. (oo) cysts in green leafy vegetables marketed in the city of Valencia (Spain) combining parasitological methods, two real-time qPCRs and light microscopy. An experimental field study was conducted on 129 vegetable samples, 64 from conventional farms and 65 from ecological (organic) farms. The samples were washed with water, and the resulting solution after removing the vegetables, was subjected to 24-hour sedimentation. The concentrated sediment was used for the search for protozoa. A positive result by both real-time PCRs, or a positive result by one qPCR and confirmation by microscopy was established as a positivity criterion. Giardia duodenalis was detected in 23.0 % of the samples, and Cryptosporidium spp. in 7.8 %. G. duodenalis (41.5 %) and Cryptosporidium spp. (20.0 %) were more frequent in ecological crops. The high level of contamination detected in organic vegetables may be due to the type of fertilizers and the quality of the water used for their irrigation and reinforces the need to take extreme hygiene measures in vegetables that are consumed raw

ICT tools for tackling bullying in schools: an analysis and opportunities

[EN] Bullying is defined as the act of repeatedly and intentionally causing harm to another person who feels helpless (i.e., the victim) against the perpetrator or group of perpetrators. These series of acts of physical and/or psychological violence have been reported to cause a negative impact on the victim on several aspects of his/her well-being and daily life. Recently, multidisciplinary research teams have noticed the potential of ICT tools as catalysts for fighting bullying in schools. A range of ICT-based tools such as elearning systems, intelligent tutoring systems, gamified applications, analytics, or even artificial intelligence have been proposed as technological assets against bullying. In this paper, we identify, categorize, and analyse the use of these ICT tools against bullying under a wide range of criteria such as their role (e.g., prevention, mitigation, detection, etc.), the potential investment that is required, the target age of users, and strengths and weaknesses of each tool. Then, we discuss on potential areas of expansion for the use of ICT tools in the fight bullying, and we identify new potential areas of research.This work has been partially funded by the Generalitat Valenciana (GV/2019/012).Alberola Oltra, JM.; Sanchez-Anguix, V.; Soto-González, MD.; Molines Borrás, S.; Monfort Torres, G.; Díaz Novillo, S. (2020). ICT tools for tackling bullying in schools: an analysis and opportunities. IATED. 4662-4667. https://doi.org/10.21125/inted.2020.1289S4662466

Association of fruits and vegetables consumption and related-vitamins with inflammatory and oxidative stress markers in prediabetic individuals

Abstract\ud \ud Background\ud Dietary guidelines of 5 servings per day of fruits and vegetables (FV) offer a reasonable amount of vitamins to control organic processes, which may contribute to a favorable cardiometabolic profile. This study aimed at investigating whether the intake of the FV group as well as pro-vitamin A carotenoids and vitamins C and E were associated with circulating markers of oxidative stress, inflammation and insulin resistance in Brazilians individuals at cardiometabolic risk.\ud \ud \ud Methods\ud This cross-sectional study included 205 individuals screened for diabetes prevention program in a healthcare center from the School of Public Health, University of São Paulo, conducted in 2008. Possible associations of consumption of FV group, as well as pro-vitamin A carotenoids and vitamins C and E, with circulating markers of oxidative stress (superoxide dismutase – SOD and oxidized LDL – oxLDL), inflammation (C reactive protein, TNF-α and adiponectin) and insulin resistance (HOMA-IR) were investigated. Pearson correlation coefficient, ANOVA and multiple linear regression were employed.\ud \ud \ud Results\ud The sample (64.7% women) had a mean age of 54.1 ± 12.7 years and body mass index of 30.7 ± 5.7 kg/m2. Dietary, physical activity, anthropometric and laboratory data were obtained. Participants consumed a mean of 3.8 servings/day of FV; their FV intake was categorized into three groups: <2.5, 2.5-5.0 and >5.0 servings/day. Significant trends for lower waist circumference (103.4 ± 13.6 vs. 100.1 ± 12.2 vs. 98.2 ± 12.7 cm, p-trend <0.05) and higher adiponectin concentrations (10.4 ± 1.8 vs. 11.9 ± 1.9 vs. 13.6 ± 2.1 ng/mL, p-trend <0.05) were detected across categories. Associations between SOD concentrations (β 0.172 [0.110-0.688]) with FV consumption and between oxLDL concentrations with vitamins C (β -0.333 [−2.568 – -0.218]) and E (β -0.354 [−1.131– -0.110]) intakes, adjusted for age, gender, BMI, saturated fat intake, smoking and physical activity were found. Similar results were observed for the associations between oxLDL and FV intake, but significance disappeared adding adjustment for saturated fat, smoking and physical activity.\ud \ud \ud Conclusion\ud Our data suggest that the intake of FV or selected vitamins may be useful for identifying the oxidative stress and inflammation involved in the genesis of cardiometabolic diseases and for motivating at-risk patients for changing dietary habits.This work was supported by a grant from The São Paulo Foundation for\ud Research Support – FAPESP and from Coordination for Improvement of\ud Graduate Personnel – CAPES (Brazil)

Índice de Qualidade da Dieta Revisado para população brasileira

El Índice de Calidad de la Dieta Revisado es un indicador de calidad de la dieta desarrollado cónsono con las recomendaciones nutricionales vigentes. Los datos dietéticos fueron provenientes del estudio de base poblacional Inquérito de Saúde e Alimentação (ISA - Pesquisa de Salud y Alimentación)-Capital-2003. El Índice contiene 12 componentes, siendo nueve fundamentados en los grupos de alimentos de la Guía Alimentaria Brasileña de 2006, cuyas porciones diarias son expresadas en densidad energética; dos nutrientes (sodio y grasa saturada); y Gord_AA (calorías provenientes de grasa sólida, alcohol y azúcar de adición). El Índice de Calidad de la Dieta Revisado propicia medir varios factores de riesgos dietéticos para enfermedades crónicas, permitiendo, simultáneamente, evaluar y monitorear la dieta en nivel individual o poblacional.O Índice de Qualidade da Dieta Revisado é um indicador de qualidade da dieta desenvolvido consoante com as recomendações nutricionais vigentes. Os dados dietéticos foram provenientes do estudo de base-populacional, Inquérito de Saúde e Alimentação (ISA)-Capital-2003. O Índice contém 12 componentes, sendo nove fundamentados nos grupos de alimentos do Guia Alimentar Brasileiro de 2006, cujas porções diárias são expressas em densidade energética; dois nutrientes (sódio e gordura saturada); e Gord_AA (calorias provenientes de gordura sólida, álcool e açúcar de adição). O Índice de Qualidade da Dieta Revisado propicia mensurar variados fatores de riscos dietéticos para doenças crônicas, permitindo, simultaneamente, avaliar e monitorar a dieta em nível individual ou populacional.The revised version of the Brazilian Healthy Eating Index is an indicator of dietary quality developed according to current nutritional recommendations. Dietary data were obtained from a population-based survey, the 2003 Inquérito de Saúde e Alimentação (ISA - Health and Diet Survey)-Capital. The Revised Index consists of 12 components: nine food groups included in the 2006 Brazilian Dietary Guidelines, in which daily portions are expressed in terms of energy density; two nutrients (sodium and saturated fats), and SoFAAS (calories from solid fat, alcohol and added sugar). The Revised Brazilian Healthy Eating Index allows for the measurement of dietary risk factors for chronic diseases, evaluating and monitoring the diet at both individual and population levels