Lotus corniculatus L. is the most important agricultural species in the genus Lotus and is the most widely distributed Lotus species worldwide. L. corniculatus genotypes form complex groups that are difficult to recognize both morphologically and biochemically. Given the extensive and expensive process of isolating Simple Sequence Repeats (SSR, also called microsatellites), the possibility of using microsatellites already identified in related species is highly attractive. The aim of this work was the identification and validation of transferable microsatellite markers in L. corniculatus, and using those markers to study the genetic variability among four cultivars. Each cultivar of L. corniculatus was represented by 15 genotypes. Ten microsatellite markers were evaluated, and from those, four were selected based on their discriminative values observed among cultivars. We detected 29 alleles for the four markers, and there was an average of 7.25 alleles per locus. The marker TM0197 had the fewest number of alleles (5) and TM0083 had the highest number of alleles (10). The polymorphic information content (PIC) for the selected markers varied from 0.19 to 0.35, and the markers were therefore classified as highly informative. Based on the markers, we found high variability between individuals of the same cultivar. The use of transferable microsatellite markers could be useful to differentiate individuals at a relatively low cost, showing a great potential for use in breeding programs.

Lotus corniculatus L. es la especie de mayor importancia agronómica en el género Lotus y la de mayor distribución a nivel mundial. Los genotipos de L. corniculatus forman grupos genéticos complejos y difíciles de reconocer tanto morfológica como bioquímicamente. Dado el proceso largo y costoso de aislar secuencias simples repetidas (SSR, microsatélites), resulta altamente atractiva la posibilidad de utilizar microsatélites ya identificados en una especie relacionada. El objetivo del trabajo fue la identificación y validación de marcadores microsatélites transferibles y la caracterización molecular de cuatro cultivares. Cada cultivar de L. corniculatus fue representado por 15 genotipos. Se evaluaron 10 marcadores microsatélites provenientes de L. japonicus L. Se comprobó la transferibilidad de marcadores de L. japonicus en L. corniculatus. Se detectaron un total de 29 alelos para los 4 microsatélites trasferibles seleccionados, mientras que el promedio de alelos por locus fue de 7,25. El marcador TM0197 fue el que menos alelos presentó (5 alelos) y el TM0083 el que más presentó (10 alelos). El contenido de información polimórfica (PIC) para los marcadores seleccionados varió entre 0,19 y 0,35 calificando en el rango de altamente informativos. Se comprobó una alta variabilidad entre individuos del mismo cultivar. El empleo de marcadores microsatélites puede ser de gran utilidad para diferenciar individuos a un bajo costo, presentando un gran potencial para su uso en mejoramiento.

The aim of the study was to evaluate the antifungal activity of extracts of 10 plant species used in traditional Uruguayan medicine against the phytopathogenic fungus Alternaria spp. The plants were selected on the basis of their reported ethnobotanical uses. Aqueous, saline buffer and acid extracts of different plant species were screened in vitro for their antifungal activity against Alternaria spp. For the antifungal evaluation we used a microspectrophotometric assay. Minimal inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the extracts were determined. Three solvents were assayed on different tissues of the plants and among the 29 evaluated extracts, 31% of the extracts inhibited growth, similar to the effects of a chemical fungicide. Acid extracts of the plants were more effective than the aqueous or buffer extracts against Alternaria spp. The MIC values of the extracts were determined ranging between 1.25 and 25 µg mL-1. The MFC values of the extracts ranged between 1.25 µg mL-1 (Rosmarinus officinalis L.) and 10 µg mL-1 (Cynara scolymus L.). MICs and MFCs values obtained from leaves (Salvia officinalis L. and R. officinalis) and seeds extracts (Salvia sclarea L.) were quite comparable to values obtained with the conventional fungicide captan (2.5 µg mL-1). The extracts of Salvia sclarea, S. officinalis and R. officinalis could be considered as potential sources of antifungal compounds for treating diseases in plants. These extracts showed maximum activity, even at very low concentrations, and the same fungicide effects as chemical fungicide. We conclude from this that these extracts exhibit amazing fungicidal properties that support their traditional use as antiseptics.

El objetivo de este trabajo fue evaluar la actividad antifúngica de extractos vegetales de 10 especies utilizadas en la medicina tradicional uruguaya contra el hongo fitopatógeno Alternaria spp. Las plantas fueron seleccionadas en base a usos etnobotánicos reportados. Se evaluó in vitro la actividad antifúngica de tres tipos de extractos, acuoso, tampón salino de extracción y ácido, de las diferentes especies vegetales. Para la evaluación de la actividad antifúngica se usó un ensayo microespectrofotométrico. Para los extractos se determinaron los parámetros de mínima concentración inhibitoria (MIC) y mínima concentración fungicida (MFC). Los tres solventes utilizados fueron ensayados en diferentes órganos vegetales; de los 29 extractos evaluados, el 31% presentó una inhibición del crecimiento en Alternaria spp. similar a la de un fungicida químico. Los extractos ácidos fueron los más efectivos. Los valores de MIC obtenidos variaron entre 1,25 y 25,0 µg mL-1. Para MFC se observaron valores en el rango de 1,25 µg mL-1 (Rosmarinus officinalis L.) y 10,0 µg mL-1 (Cynara scolymus L.). Los valores de MIC y MFC de extractos obtenidos de hojas (Salvia officinalis L. y R. officinalis) y semillas (Salvia sclarea L.) fueron comparables a los valores obtenidos con el fungicida convencional captan (2,5 µg mL-1). Los extractos de S. sclarea, S. officinalis y R. officinalis pueden ser fuentes potenciales de compuestos antifúngicos para el tratamiento de enfermedades en plantas. Estos extractos mostraron máxima actividad inhibitoria a bajas concentraciones, teniendo un comportamiento similar a la del fungicida químico. Estos resultados permiten concluir que estos extractos presentan sorprendentes propiedades fungicidas, fundamentando su uso tradicional como antisépticos.

Fostering translational research of advanced therapies has become a major priority of both scientific community and national governments. Advanced therapy medicinal products (ATMP) are a new medicinal product category comprising gene therapy and cell-based medicinal products as well as tissue engineered medicinal products. ATMP development opens novel avenues for therapeutic approaches in numerous diseases, including cancer and neurodegenerative and cardiovascular diseases. However, there are important bottlenecks for their development due to the complexity of the regulatory framework, the high costs and the needs for good manufacturing practice (GMP) facilities and new end-points for clinical experimentation. Thus, a strategic cooperation between different stakeholders (academia, industry and experts in regulatory issues) is strongly needed. Recently, a great importance has been given to research infrastructures dedicated to foster translational medicine of advanced therapies. Some ongoing European initiatives in this field are presented and their potential impact is discussed.

La promozione della ricerca traslazionale nel campo delle terapie avanzate è avvertita sempre di più come una priorità sia dalla comunità scientifica che dai governi nazionali. I prodotti medicinali per terapie avanzate (PMTA) sono una nuova categoria di farmaci e comprendono tre tipologie di prodotti: prodotti per terapia cellulare somatica, prodotti per terapia genica e prodotti di ingegneria tissutale. Lo sviluppo dei PMTA apre nuove strade per approcci terapeutici a varie malattie incluso il cancro, le malattie neurodegenerative e le malattie cardiovascolari. Tuttavia questo processo è ostacolato dalla complessità della cornice regolatoria, dai costi elevati e dalla necessità di produzione GMP (good manufacturing practice, buona pratica di fabbricazione) e di nuovi end-point per la sperimentazione clinica. Quindi è necessario che si stabilisca una cooperazione strategica tra i differenti "stakeholders" (mondo accademico, industria ed esperti in campo regolatorio). Recentemente ha assunto una grande importanza lo sviluppo di infrastrutture di ricerca dedicate alla promozione della ricerca traslazionale nel campo delle terapie avanzate. In questo lavoro vengono presentate alcune iniziative europee in questo ambito e viene discusso il loro impatto potenziale.

The bread making quality of wheat (Triticum aestivum L.) depends on the quality and quantity of the gluten proteins -glutenins and gliadins- which constitute 10 to 14% of the grain protein content. Various quantitative parameters, such as the total content of proteins of the flour, content of polymeric proteins present in the grain and the glutenin/gliadin ratio are related to baking quality. This work presents the characterization of high molecular weight glutenin subunits (HMW-GS) by sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis (SDS-PAGE) of 22 Uruguayan cultivars and breeding lines. 14 HMW-GS subunits were observed corresponding to 10 allele combinations. On the other hand, it is proposed a simple method to group and quantify the main storage proteins of bread wheat by SE-HPLC (Size Exclusion High Performance Liquid Chromatography). The chromatograms obtained showed an excellent separation of the endosperm proteins in three main peaks corresponding to polymeric, monomeric and non gluten proteins, respectively. The integrated areas of the chromatograms as well as the presence of subunits of the cultivars were analyzed. The genotypes analyzed showed quantitative differences in the quantity of glutenin/gliadin solubles in SDS and in the proportion of polymeric and monomeric proteins. The cultivar INIA Gorrión of good baking quality having the alleles Glu-D1 5+10 showed a lower percentage of SDS-soluble polymeric proteins and higher insoluble polymeric proteins than the experimental line LE 2265, which has the alleles Glu-D1 2+12.

La calidad del trigo para pan (Triticum aestivum L.) depende de la calidad y cantidad de las proteínas del gluten -gluteninas y gliadinas- las cuales constituyen 10 a 14% de las proteínas del grano. Varios parámetros cuantitativos, como el contenido total de proteínas de la harina, el contenido de proteínas poliméricas presentes en el grano y la proporción de gluteninas y gliadinas, están relacionados a la calidad panadera. En este trabajo se presenta la caracterización de las gluteninas de alto peso molecular (G-APM) en geles de poliacrilamida en SDS (SDS-PAGE) de 22 cultivares y líneas de mejoramiento de Uruguay. Fueron observadas 14 subunidades de G-APM correspondientes a 10 combinaciones alélicas. Por otra parte, se propone un método sencillo para agrupar y cuantificar las principales proteínas de reserva de trigo mediante cromatografía líquida de exclusión por tamaño de alta resolución (SE-HPLC). Los cromatogramas obtenidos mostraron una excelente separación de las proteínas del endospermo en tres picos principales correspondientes a proteínas poliméricas, monoméricas y no gluten, respectivamente. El área de integración de los cromatogramas así como la presencia de subunidades de los cultivares fueron analizados. Los genotipos estudiados mostraron diferencias cuantitativas en el contenido de gluteninas/gliadinas solubles en SDS, en la proporción de proteínas poliméricas y monoméricas. El cultivar INIA Gorrión, de buena calidad panadera y que presenta el alelo Glu-D1 5+10, mostró menor porcentaje de proteínas poliméricas solubles en SDS y mayor porcentaje de proteínas poliméricas insolubles en comparación con la línea experimental LE 2265, que tiene el par Glu-D1 2+12.