Diagnostics més precisos gràcies a noves tècniques de citogenètica molecular

Cristina Hernando ha presentat la seva tesi doctoral on evidencia la utilitat de les noves tècniques de citogenètica molecular per a la caracterització de petits desequilibris i reorganitzacions del genoma en diferents teixits. La principal aportació d'aquesta tesi ha estat l'establiment de diferents correlacions entre les anomalies cromosòmiques detectades i les característiques clíniques observades en cada pacient.Cristina Hernando ha presentado su tesis doctoral en la que evidencia la utilidad de las nuevas técnicas de citogenética molecular para la caracterización de pequeños desequilibrios y reorganizaciones del genoma en diferentes tejidos. Lla principal aportación de la tesis es el establecimiento de diferentes correlaciones entre las anomalías cromosómicas detectadas y las características clínicas observadas en cada paciente.Cristina Hernando shows in her doctoral thesis the utility of the new technologies in Molecular Cytogenetics, which is helpful to the characterization of imbalance and genome reorganization in different tissues. This work shows new correlations between genotype- fenotype which allow to give a genetic advice more accurate, surely the most important contribution of this work

Caracterización de anomalías cromosómicas en diagnóstico prenatal y postnatal mediante técnicas de citogenética molecular

Consultable des del TDXTítol obtingut de la portada digitalitzadaLas anomalías cromosómicas están implicadas en la aparición de muchas enfermedades hereditarias, siendo también la principal causa de retraso mental y de pérdidas gestacionales. La pérdida, ganancia o reordenación de fragmentos cromosómicos de un tamaño similar, puede tener distintas consecuencias dependiendo del número y función de los genes que contengan. Los métodos clásicos de bandeo cromosómico han sido durante muchos años la única herramienta para la detección de anomalías cromosómicas tanto numéricas como estructurales. Gracias a estas técnicas se han podido identificar muchas anomalías cromosómicas asociadas a diferentes malformaciones congénitas, síndromes o cánceres colaborando eficazmente en la comprensión de la etiología de dichas patologías. No obstante, las alteraciones cromosómicas de un tamaño < 3-5 Mb o las reorganizaciones complejas son muy difíciles de identificar mediante las técnicas de bandas convencionales. Desde su inicio, hacia 1980, las técnicas de citogenética molecular basadas en la hibridación in situ fluorescente (FISH) y sus variantes tecnológicas, entre ellas la Hibridación Genómica Comparada (CGH) i la FISH-Multicolor (M-FISH), han permitido la detección e identificación precisa de un gran número de anomalías cromosómicas, como las microdeleciones, las reorganizaciones cromosómicas complejas y las reorganizaciones crípticas, que hasta ese momento pasaban desapercibidas. El objetivo principal de este trabajo ha sido profundizar en el papel que desempeñan regiones cromosómicas específicas, que al alterarse, originan fenotipos concretos mediante la aplicación conjunta de diferentes técnicas de citogenética molecular (CGH y FISH) y de citogenética convencional. Hoy en día, se conocen muy pocas regiones cromosómicas cuya alteración, por pérdida o duplicación, esté asociada a un síndrome clínico bien definido. Distintos estudios, incluido el presente, muestran que tanto la CGH, como la M-FISH, son dos técnicas que contribuyen eficazmente en la identificación de regiones cromosómicas específicas asociadas a fenotipos concretos. Esta información es de suma importancia para los biólogos moleculares como indicación de qué regiones del genoma deben considerar como «dianas» para identificar los genes implicados. En esta tesis además de la puesta a punto u optimización de las técnicas de M-FISH, CGH y HR-CGH para su aplicación al diagnóstico prenatal y postnatal, se han determinado las limitaciones tanto de las propias técnicas como de los softwares empleados. Ello nos ha permitido establecer nuevas correlaciones genotipo-fenotipo gracias a la identificación de 27 monosomías y trisomías autosómicas y gonosómicas, 15 cromosomas marcadores supernumerarios y 9 reorganizaciones complejas y/o crípticas. Al mismo tiempo, se ha establecido un protocolo para la identificación de marcadores cromosómicos mediante la utilización combinada de las técnicas de citogenética convencional, FISH y CGH. Por último, el análisis de los 77 puntos de rotura implicados en las anomalías cromosómicas estudiadas ha revelado que no se producen al azar y que fundamentalmente afectan a los cromosomas: 2, 7, 9, 15, 16, 18, X e Y. Las bandas más afectadas han sido: 9p23, 11q22.2, 14q11.2, 15q11.2, 15q15, 16p11.2, 18q22, 22q11.2, Xp22.3, Xq21.2, Yp11.3 e Yq12, evidenciando que las roturas cromosómicas se producen en bandas cromosómicas claras (80%) que corresponden a regiones del genoma con un mayor contenido genético. El análisis de estos puntos revela que coinciden en un 90% con regiones donde se ha descrito duplicaciones segmentarias. Este último hallazgo es de suma importancia y abre un nuevo campo de investigación. En resumen, esta tesis muestra la utilidad y aplicabilidad de la CGH y M-FISH, tanto en Diagnóstico Prenatal como Postnatal, al identificar tanto pequeños desequilibrios como reorganizaciones complejas y/o crípticas permitiendo establecer nuevas correlaciones genotipo-fenotipo que facilitaran en un futuro un consejo genético mucho más preciso.Chromosomal abnormalities are involved in the appearance of many hereditary diseases representing the principal factor of mental retardation as well as miscarriages. The loss, gain or redistribution of chromosomal fragments with a similar size may have different consequences depending on the number and function of the respective genes. During lots of years conventional chromosomal banding techniques had been applied as a unique tool in order to detect numerical and structural chromosomal abnormalities. Thanks to those techniques a significant number of chromosome-related abnormalities associated with congenital malformations, diverse syndromes and cancer forms could be indeed identified consolidating on this way, even a better comprehension of the etiology of these pathologies. However, the chromosomal alterations with a size of < 3-5 Mb or complex reorganizations are rather difficult to be identified by conventional banding techniques. Since 1980 molecular cytogenetic techniques based on a fluorescence in situ hybridization (FISH) with its technologic variants as for instance the Comparative Genomic Hybridization (CGH) and the multicolour-FISH (M-FISH) made possible the exact detection and identification of numerous chromosomal abnormalities as there are microdeletions, complex chromosomal reorganizations as well as cryptic reorganizations which until that moment could not be recognized. The main objective of this work consisted of focussing the specific chromosomal regions which cause in case of alteration concrete phenotypes by means of a combined application of different molecular cytogenetic and conventional techniques (CGH and FISH). Nowadays we dispose of a very limited know how about chromosomal regions of which an alteration by deletion or duplication is associated with a well defined clinical syndrome. Different studies including the present one had showed that both procedures CGH and M-FISH are techniques able to identify efficiently specific chromosomal regions associated with specific phenotypes. This information is a quite important for molecular biologists as it reveals which regions of a genome can be considered as key regions within the definition of implicated genes. With this doctoral thesis, besides the improvement and/or optimisation of M-FISH, CGH and HR-CGH techniques relevant for prenatal and postnatal diagnosis, limitations had been determined on the one hand regarding the same techniques and on the other hand with respect to the software. That circumstance allowed us indeed to establish new genotype-phenotype correlations due to the identification of 27 autosomal and gonosomal monosomies and trisomies, 15 supernumerary marker chromosomes and 9 complex and/or cryptic reorganizations. At the same time a protocol could be established for the identification of supernumerary marker chromosomes by the combined application of the conventional cytogenetic techniques FISH and CGH. Finally the analysis of the 77 breakpoints implicated in the investigated chromosomal abnormalities unveiled that they had not been brought out accidentally and that they fundamentally affect the chromosomes: 2, 7, 9, 15, 16, 18, X and Y. On the other hand the bands more affected were 9p23, 11q22.2, 14q11.2, 15q15, 16p11.2, 18q22, 22q11.2, Xp22.3, Xq21.2, Yp11.3 and Yq12, showing that chromosome breaks are mainly located on clear chromosome bands (80%) corresponding to genome regions with a bigger genetic content. The analysis of these points reveals that a 90 % of them coincide with regions in which segmental duplications had been described. This last recognition represents an outstanding circumstance and initiates in a future a new serie of investigations. Summing up this doctoral thesis shows the utility and applicability of CGH and M-FISH as prenatal and postnatal diagnosis identifying as well slighter deviations as complex and cryptic reorganisations. It will equally allow us to establish new genotypic and phenotypic correlations making possible in a future a more exact genetic assessment

Topics on “Libra” electronic nose for fruit quality

Quartz crystal microbalances (QCM) are piezoelectric devices. To turn a quartz crystal into a chemical sensor it is necessary to coat it with a layer of a material capable of capturing molecules from the environment. When a mass is absorbed or placed onto the quartz crystal surface, the oscillation frequency changes decreasing in proportion to the amount of mass (Di Natale et al. 1997). The ability to control a QCM´s selectivity by applying different coatings is an important feature, and makes this sensor type extremely versatile. However, the coating of QCM is, ironically, their greatest drawback. Batch-to-batch variability in the manufacturing leads to inadequate reproducibility(Sarig 2000). Indeed, the response of sensors depends on numerous factors that may be difficult to control, such as the temperature and the humidity of the carrier gas. All these factors cause changes in the selectivity of sensors affecting the reproducibility of measurements. The detection threshold of the human nose is typically between 1000 ppm (10-6)and <1ppt (10-12). Therefore, the absolute detection threshold of a gas sensor should be very low. For a QCM e-nose, only a few molecules are required to react with the sensitive elements leading to sensitivities close to the ppm or tenth of a ppm range as measured in the vapour phase (Mielle 1996) . Despite previous studies using electronic nose, no QCM calibration statement has been established in relation to the sensitivity needed in the sensors for organoleptic quality measurement in pear or in other fruit. Present work will show the possibilities of using this equipment as a non destructive method to evaluate fruit quality. The work will be developed in different steps or topics to get a better understanding

In Vitro and In Vivo Digestion of Persimmon and Derived Products: A Review

[EN] The link between nutrition and health has focused on the strategy of diet-based programs to deal with various physiological threats, such as cardiovascular disease, oxidative stress, and diabetes. Therefore, the consumption of fruits and vegetables as a safeguard for human health is increasingly important. Among fruits, the intake of persimmon is of great interest because several studies have associated its consumption with health benefits due to its high content of bioactive compounds, fiber, minerals, and vitamins. However, during digestion, some changes take place in persimmon nutritional compounds that condition their subsequent use by the human body. In vitro studies indicate different rates of recovery and bioaccessibility depending on the bioactive compound and the matrix in which they are found. In vivo studies show that the pharmacological application of persimmon or its functional components, such as proanthocyanidins, can help to prevent hyperlipidemia and hyperglycemia. Thus, persimmon and persimmon derived products have the potential to be a fruit recommended for diet therapy. This review aims to compile an updated review of the benefits of persimmon and its derived products, focusing on the in vitro and in vivo digestibility of the main nutrients and bioactive compounds.Grant RTA2017-00045-C02-02 funded by MCIN/AEI/ 10.13039/501100011033 and, by ERDF A way of making Europe.González, CM.; Hernando Hernando, MI.; Moraga Ballesteros, G. (2021). In Vitro and In Vivo Digestion of Persimmon and Derived Products: A Review. Foods. 10(12):1-15. https://doi.org/10.3390/foods10123083S115101

Influence of ripening stage and de-astringency treatment on the production of dehydrated persimmon snacks

[EN] BACKGROUND Seasonal persimmon (Diospyros kakiL.) crops have steadily increased in Spain; this has been linked to a significant increase in the postharvest production waste. Therefore, development of valorized products is of great interest. In this study, a hot air-drying technique was used to obtain persimmon snacks. Slices from astringent and non-astringent persimmons (submitted to de-astringency treatment) at three different ripening stages were dried at 40 and 60 degrees C to reach 15 +/- 3% water content. RESULTS After the drying treatment, dehydrated samples were harder, turned into a more orange hue angle, and had a reduced soluble tannin content. Dehydrated samples obtained from the astringent fruit at the most advanced ripening stage had similar soluble tannin content as the samples obtained from non-astringent fruit, especially at 60 degrees C. Besides, a high correlation was observed between the level of astringency perceived by consumers and the decrease of soluble tannin content. Although, in the first ripening stage, consumers preferred the snacks obtained from non-astringent fruits; in the last ripening stage, snacks produced from astringent fruits were equally accepted than the non-astringent ones. CONCLUSION Therefore, well-accepted persimmon snacks are obtained from both astringent and non-astringent fruits when advanced ripening stages of persimmon are used.The authors thank the Ministerio de Ciencia, Innovacion y Universidades for the financial support given throughout Project RTA2017-00045-C02-02. They would also like to thank Phillip Bentley for assistance in correcting the English manuscript.González, CM.; Hernando Hernando, MI.; Moraga Ballesteros, G. (2021). Influence of ripening stage and de-astringency treatment on the production of dehydrated persimmon snacks. Journal of the Science of Food and Agriculture. 101(2):603-612. https://doi.org/10.1002/jsfa.10672603612101

Designing a Clean Label Sponge Cake with Reduced Fat Content

[EN] The fat in a sponge cake formulation was partially replaced (0%, 30%, 50%, and 70%) with OptiSolTM5300.This natural functional ingredient derived from flax seeds, rich in fiber and alpha-linoleic acid, provides a natural substitute for guar and xanthan gums, avoiding E-numbers on labels. The structure and some physicochemical properties of the formulations were examined, sensory analysis was conducted and changes in starch digestibility due to adding this ingredient were determined. Increasing quantities of OptiSolTM5300 gave harder cakes, with less weight loss during baking, without affecting the final cake height. There were no significant differences (P > 0.05) in texture, flavor and overall acceptance between the control and the 30% substitution cake, nor in the rapidly digestible starch values. Consequently, replacing up to 30% of the fat with OptiSolTM5300 gives a new product with health benefits and a clean label that resembles the full-fat sponge cake.The authors are grateful to INIA for financial support through the BERRYPOM- Adding value to fruit processing waste: innovative ways to incorporate fibers from berry pomace in baked and extruded cereal-based foods project included in the ERA-NET - SUSFOOD program.Eslava-Zomeño, C.; Quiles Chuliá, MD.; Hernando Hernando, MI. (2016). Designing a Clean Label Sponge Cake with Reduced Fat Content. Journal of Food Science. 81(10):2352-2359. doi:10.1111/1750-3841.1344623522359811

Optimization of a portable NIR device for the optical supervision of milk coagulation process

The coagulation of milk is the fundamental process in cheese-making, which is based on a gel formation as consequence of physicochemical changes taking place in the casein micelles. Monitoring the whole process of milk curd formation is a dedicated process for dairy researchers and cheese companies. In addition to advances in composition-based applications by means of NIR spectroscopy, researchers are pursuing dynamic applications that show promise especially with regard to tracking a sample in situ during processing The objective of this work is to propose an original portable NIR equipment to supervise the milk coagulation process. The experiments have been carried out on sheep and goat milk, by immersion of the probe directly in the liquid and acquiring spectrum each 1 minute during the 30 minutes of coagulation process. The increasing values of transflected light registered allow identifying, based on PCA analysis, the different kinetics that occur along the gel formation and the time to reach the optimal gel firmness to cut the cur

Envases inteligentes que mejoran los zumos

Las funciones principales de los envases son la contención del alimento y su protección y preservación de la contaminación externa, contribuyendo también a la comodidad, el marketing y la comunicación con el consumidor. La alta penetración de tecnologías que posibilitan el acceso a la información en todos los ámbitos y en tiempo real, junto con el abaratamiento de las tecnologías que permiten la actuación y monitorización sobre los envases individuales, son las razones tecnológicas que se combinan con otras razones legislativas y sociológicas, como las demandas y las exigencias de unos consumidores formados y una normativa cada vez más restrictiva, para hacer suponer que los envases activos e inteligentes serán muy pronto realidades frecuentes en nuestras estanterías. En este trabajo se presentan algunas de las soluciones que ya existen en el mercado y otras que estarán disponibles en un futuro muy próximo para incorporar/integrar en los envases tradicionalmente empleados en productos líquidos como zumos y batidos de frutas y hortalizas frescos, elementos activos e inteligentes que jugarán un papel importante en la diferenciación de estos productos en los mercados de los países europeos

Poscosecha de Precisión en los Albores del siglo XXI

Según la FAO (Gustavsson et al., 2011) todos los años un tercio de los alimentos producidos para consumo humano se pierden o desperdician. En su informe de 2011 ‘Pérdidas y desperdicio de alimentos en el mundo’, este organismo cifra en 1.300 millones de toneladas los alimentos que no llegan al consumidor final. Las altas pérdidas que se producen también en los procesos poscosecha en forma de derrames, de disminución de la calidad y/o acortamiento de la vida útil de frutas y hortalizas justifican la necesidad de invertir en el manejo y procedimientos poscosecha antes de pensar incluso en el incremento de las superficies de cultivo (Yahla, 2009)