ABSTRACT
One alternative for adapting viticulture to high temperatures and the scarcity of water is the development of new varieties adapted to such conditions. This work describes six new genotypes, derived from "Monastrell" × "Cabernet Sauvignon" (MC16, MC19, MC72, MC80) and "Monastrell" × "Syrah" (MS104, MS49) crosses, grown under deficit irrigation and rainfed conditions in a semi-arid wine-producing area (Murcia, southeastern Spain). The effect of genotype, year, and irrigation treatment on the phenological, productiveness, morphological, and grape quality data was evaluated. The study material was obtained and selected as part of a breeding program run by the Instituto Murciano de Investigación y Desarollo Agrario y Medioambiental (IMIDA). The results obtained show that under rainfed conditions, the values for productive variables decreased, while those referring to the phenolic content increased. Notable variation in the parameters evaluated was also seen for the different genotypes studied. The behavior of the genotypes MC80 and MS104 under rainfed conditions was noteworthy. In addition to maintaining very adequate yields, phenolic contents, must pH, and total acidity values, MC80 fell into the best 'phenolic quality group' and MS104 returned a low º°Baumé value, ideal for the production of low-alcohol-content wines. These genotypes could favor the development of sustainable quality viticulture in dry and hot areas.
ABSTRACT
The need to develop an environmentally friendly, sustainable viticulture model has led to numerous grapevine improvement programmes aiming to increase resistance to downy and powdery mildew. The success of such programmes relies on the availability of protocols that can quantify the resistance/susceptibility of new genotypes, and on the existence of molecular markers of resistance loci that can aid in the selection process. The present work assesses the degree of phenotypic resistance/susceptibility to downy and powdery mildew of 28 new genotypes obtained from crosses between "Monastrell" and "Regent." Three genotypes showed strong combined resistance, making them good candidates for future crosses with other sources of resistance to these diseases (pyramiding). In general, laboratory and glasshouse assessments of resistance at the phenotype level agreed with the resistance expected from the presence of resistance-associated alleles of simple sequence repeat (SSR) markers for the loci Rpv3 and Ren3 (inherited from "Regent"), confirming their usefulness as indicators of likely resistance to downy and powdery mildew, respectively, particularly so for downy mildew.
ABSTRACT
Wound healing is a biological process directed to the restoration of tissue that has suffered an injury. An important phase of wound healing is the generation of a basal epithelium able to wholly replace the epidermis of the wound. A broad range of products derived from fibroin and sericin from Bombyx mori silk are used to stimulate wound healing. However, so far the molecular mechanism underlying this phenomenon has not been elucidated. The aim of this work was to determine the molecular basis underlying wound healing properties of silk proteins using a cell model. For this purpose, we assayed fibroin and sericin in a wound healing scratch assay using MDA-MB-231 and Mv1Lu cells. Both proteins stimulated cell migration. Furthermore, treatment with sericin and fibroin involved key factors of the wound healing process such as upregulation of c-Jun and c-Jun protein phosphorylation. Moreover, fibroin and sericin stimulated the phosphorylation of ERK 1/2 and JNK 1/2 kinases. All these experiments were done in the presence of specific inhibitors for some of the cell signalling pathways referred above. The obtained results revealed that MEK, JNK and PI3K pathways are involved in fibroin and sericin stimulated cells migration. Inhibition of these three kinases prevented c-Jun upregulation and phosphorylation by fibroin or sericin. Fibroin and sericin were tested in the human keratinocyte cell line, HaCaT, with similar results. Altogether, our results showed that fibroin and sericin initiate cell migration by activating the MEK, JNK and PI3K signalling pathways ending in c-Jun activation.
Subject(s)
Cell Movement/physiology , Fibroins/physiology , Proto-Oncogene Proteins c-jun/metabolism , Sericins/physiology , Silk/chemistry , Up-Regulation/physiology , Animals , Bombyx , Cell Line , Humans , Phosphorylation , Protein Kinase Inhibitors/pharmacology , Protein Kinases/metabolismABSTRACT
INTRODUCCION: La leishmaniasis tegumentaria americana (LTA) es una enfermedad endémica reemergente en la provincia de Salta. Los recursos terapéuticos disponibles presentan serias limitaciones. Se han detectado numerosos casos de fallas terapéuticas debido a que el criterio de curación se basa en la evolución clínica de las lesiones.OBJETIVO: A fin de contar con un sistema adecuado para monitorear la enfermedad, se propuso optimizar una PCR en tiempo real (RT-PCR) basada en el uso de un agente intercalante y oligonucleótidos que amplifican secuencias del ADN del kinetoplasto (KADN), directamente de muestras de raspados de lesiones (en pacientes con LTA) contenidas en buffer TE (Tris-EDTA), sin extracción de ADN.METODOS: Para obtener una curva estándar (CE) se realizaron diluciones seriadas de parásitos (p) de cepas de referencia de especies que circulan en la zona. También se compararon 3 métodos de procesamiento de muestras (PM): Buffer de lisis, Insta-GeneTM Matrix (BIO-RAD) y TE. Luego se analizaron negativos para evaluar la especificidad y sensibilidad del sistema. Estas muestras provenían de raspados de lesiones cutáneas o mucocutáneas, tomados con palillos de madera en 300 μl TE.RESULTADOS: El límite de detección fue de 0,001 p/300 μl TE. La CE construida a partir de una cepa de Leishmania Viannia braziliensis mostró una pendiente de -3,40, eficiencia de amplificación de 96,66%, coeficiente de Pearson (R2) de 0,997 e intersección en la ordenada de 44,079. Al comparar los PM, el método de buffer TE fue el más eficiente. La RT-PCR desarrollada mostró un 100% de especificidad frente a muestras controles negativos y un 100% de sensibilidad frente a controles positivos.CONCLUSIONES: El sistema analizado es altamente sensible y permite detectar parásitos de Leishmania sp directamente de muestras clínicas provenientes de raspados de lesiones.
INTRODUCTION: The american tegumentary leishmaniasis (ATL) is an endemic, re-emergent disease in the Province of Salta. The therapeutic resources which are available have serious limitations. Numerous treatment failures have been detected due to the fact that the evaluation of chemotherapy is based on the clinical outcome of lesionsOBJECTIVE: In order to find a system for the correct follow-up of the disease, the study aimed at optimized the real time PCR (RT-PCR) based on intercalating agent and primers that amplify kinetoplastic DNA sequences (KDNA), directly on samples from skin lesion scrappings (from patients with ATL) contained in buffer TE, without DNA purification..METHODS: To obtain a stardard curve (SC) serial dilutions of parasites (p) were made (from reference strains of species that circulate in the region). 3 different methods of sample processing (SP) were compared: Lysis buffer, Insta-GeneTM Matrix (BIO-RAD) and TE (Tris-EDTA). The study evaluated the specificity and sensibility of the system by analyzing positive (by conventional PCR) and negative clinical samples. These samples were from cutaneous or mucocutaneous lesion scrapings, taken with toothpicks in 300 μl TE.RESULTS: The detection limit was 0.001 p/300 μl TE. The SC built with a Leishmania Viannia braziliensis strain showed a slope of -3.40, amplification efficiency of 96.66%, Pearson coefficient (R2) of 0.997 and 44.079 x-intersection. When comparing the SP, the buffer TE method was the most efficient one. With respect to positive and negative control samples, this RT-PCR showed a sensitivity and specificity of 100% respectively.CONCLUSIONS: This system is highly sensitive and allows to detect Leishmania sp parasites directly from clinical samples of lesion scrapings.
Subject(s)
Leishmaniasis , Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Diagnosis , Argentina , Public HealthABSTRACT
INTRODUCCION: La leishmaniasis tegumentaria americana (LTA) es una enfermedad endémica reemergente en la provincia de Salta. Los recursos terapéuticos disponibles presentan serias limitaciones. Se han detectado numerosos casos de fallas terapéuticas debido a que el criterio de curación se basa en la evolución clínica de las lesiones.OBJETIVO: A fin de contar con un sistema adecuado para monitorear la enfermedad, se propuso optimizar una PCR en tiempo real (RT-PCR) basada en el uso de un agente intercalante y oligonucleótidos que amplifican secuencias del ADN del kinetoplasto (KADN), directamente de muestras de raspados de lesiones (en pacientes con LTA) contenidas en buffer TE (Tris-EDTA), sin extracción de ADN.METODOS: Para obtener una curva estándar (CE) se realizaron diluciones seriadas de parásitos (p) de cepas de referencia de especies que circulan en la zona. También se compararon 3 métodos de procesamiento de muestras (PM): Buffer de lisis, Insta-GeneTM Matrix (BIO-RAD) y TE. Luego se analizaron negativos para evaluar la especificidad y sensibilidad del sistema. Estas muestras provenían de raspados de lesiones cutáneas o mucocutáneas, tomados con palillos de madera en 300 μl TE.RESULTADOS: El límite de detección fue de 0,001 p/300 μl TE. La CE construida a partir de una cepa de Leishmania Viannia braziliensis mostró una pendiente de -3,40, eficiencia de amplificación de 96,66%, coeficiente de Pearson (R2) de 0,997 e intersección en la ordenada de 44,079. Al comparar los PM, el método de buffer TE fue el más eficiente. La RT-PCR desarrollada mostró un 100% de especificidad frente a muestras controles negativos y un 100% de sensibilidad frente a controles positivos.CONCLUSIONES: El sistema analizado es altamente sensible y permite detectar parásitos de Leishmania sp directamente de muestras clínicas provenientes de raspados de lesiones.
INTRODUCTION: The american tegumentary leishmaniasis (ATL) is an endemic, re-emergent disease in the Province of Salta. The therapeutic resources which are available have serious limitations. Numerous treatment failures have been detected due to the fact that the evaluation of chemotherapy is based on the clinical outcome of lesionsOBJECTIVE: In order to find a system for the correct follow-up of the disease, the study aimed at optimized the real time PCR (RT-PCR) based on intercalating agent and primers that amplify kinetoplastic DNA sequences (KDNA), directly on samples from skin lesion scrappings (from patients with ATL) contained in buffer TE, without DNA purification..METHODS: To obtain a stardard curve (SC) serial dilutions of parasites (p) were made (from reference strains of species that circulate in the region). 3 different methods of sample processing (SP) were compared: Lysis buffer, Insta-GeneTM Matrix (BIO-RAD) and TE (Tris-EDTA). The study evaluated the specificity and sensibility of the system by analyzing positive (by conventional PCR) and negative clinical samples. These samples were from cutaneous or mucocutaneous lesion scrapings, taken with toothpicks in 300 μl TE.RESULTS: The detection limit was 0.001 p/300 μl TE. The SC built with a Leishmania Viannia braziliensis strain showed a slope of -3.40, amplification efficiency of 96.66%, Pearson coefficient (R2) of 0.997 and 44.079 x-intersection. When comparing the SP, the buffer TE method was the most efficient one. With respect to positive and negative control samples, this RT-PCR showed a sensitivity and specificity of 100% respectively.CONCLUSIONS: This system is highly sensitive and allows to detect Leishmania sp parasites directly from clinical samples of lesion scrapings.
