μ-calpain binds to lipid bilayers via the exposed hydrophobic surface of its Ca2+-activated conformation

μ- and m-calpain are cysteine proteases requiring micro- and millimolar Ca2+ concentrations for their activation in vitro. Among other mechanisms, interaction of calpains with membrane phospholipids has been proposed to facilitate their activation by nanomolar {[}Ca2+] in living cells. Here the interaction of non-autolysing, C115A active-site mutated heterodimeric human μ-calpain with phospholipid bilayers was studied in vitro using protein-to-lipid fluorescence resonance energy transfer and surface plasmon resonance. Binding to liposomes was Ca2+-dependent, but not selective for specific phospholipid. head groups. {[}Ca2+](0.5) for association with lipid bilayers was not lower than that required for the exposure of hydrophobic surface (detected by TNS fluorescence) or for enzyme activity in the absence of lipids. Deletion of domain V reduced the lipid affinity of the isolated small subunit (600-fold) and of the heterodimer (10- to 15-fold), thus confirming the proposed role of domain V for membrane binding. Unexpectedly, mutations in the acidic loop of the `C2-like' domain III, a putative Ca2+ and phospholipid-binding site, did not affect lipid affinity. Taken together, these results support the hypothesis that in vitro membrane binding of μ-calpain is due to the exposed hydrophobic surface of the active conformation and does not reduce the Ca2+ requirement for activation

μ-calpain binds to lipid bilayers via the exposed hydrophobic surface of its Ca2+-activated conformation

μ- and m-calpain are cysteine proteases requiring micro- and millimolar Ca2+ concentrations for their activation in vitro. Among other mechanisms, interaction of calpains with membrane phospholipids has been proposed to facilitate their activation by nanomolar {[}Ca2+] in living cells. Here the interaction of non-autolysing, C115A active-site mutated heterodimeric human μ-calpain with phospholipid bilayers was studied in vitro using protein-to-lipid fluorescence resonance energy transfer and surface plasmon resonance. Binding to liposomes was Ca2+-dependent, but not selective for specific phospholipid. head groups. {[}Ca2+](0.5) for association with lipid bilayers was not lower than that required for the exposure of hydrophobic surface (detected by TNS fluorescence) or for enzyme activity in the absence of lipids. Deletion of domain V reduced the lipid affinity of the isolated small subunit (600-fold) and of the heterodimer (10- to 15-fold), thus confirming the proposed role of domain V for membrane binding. Unexpectedly, mutations in the acidic loop of the `C2-like' domain III, a putative Ca2+ and phospholipid-binding site, did not affect lipid affinity. Taken together, these results support the hypothesis that in vitro membrane binding of μ-calpain is due to the exposed hydrophobic surface of the active conformation and does not reduce the Ca2+ requirement for activation

μ-Calpain binds to lipid bilayers via the exposed hydrophobic surface of its Ca2+-activated conformation

μ- and m-calpain are cysteine proteases requiring micro- and millimolar Ca2+ concentrations for their activation in vitro. Among other mechanisms, interaction of calpains with membrane phospholipids has been proposed to facilitate their activation by nanomolar [Ca2+] in living cells. Here the interaction of non-autolysing, C115A active-site mutated heterodimeric human μ-calpain with phospholipid bilayers was studied in vitro using protein-to-lipid fluorescence resonance energy transfer and surface plasmon resonance. Binding to liposomes was Ca2+-dependent, but not selective for specific phospholipid head groups. [Ca2+]0.5 for association with lipid bilayers was not lower than that required for the exposure of hydrophobic surface (detected by TNS fluorescence) or for enzyme activity in the absence of lipids. Deletion of domain V reduced the lipid affinity of the isolated small subunit (600-fold) and of the heterodimer (10- to 15-fold), thus confirming the proposed role of domain V for membrane binding. Unexpectedly, mutations in the acidic loop of the ‘C2-like' domain III, a putative Ca2+ and phospholipid-binding site, did not affect lipid affinity. Taken together, these results support the hypothesis that in vitro membrane binding of μ-calpain is due to the exposed hydrophobic surface of the active conformation and does not reduce the Ca2+ requirement for activatio

Incidencia de la aplicación de las 5 S en el servicio de atención del área de laboratorio de la clínica Madre de Cristo. Trujillo, 2017

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Three New Promising Highly Productive Sugarcane Clones for Farming and the Industry

Context: At present, high-potential agricultural and industrial sugarcane clones are critical at the beginning of the harvest to replace the cultivars that have reduced their productivity after years of exploitation and resistance to the major pests and diseases that attack the crop. Aim: To evaluate the agro-industrial potential of three new promising sugarcane clones in dryland. Methods: A randomized experimental block design with five treatments and three replicas was used. The treatments were three promising clones (C13-369, C13-370 and C13-383), and two commercial cultivars (C86-12 y C1051-73) used as controls. The agro-botanical characteristics and crop yields, along with the juice quality parameters at the beginning of the harvest (December), at 12 months of plant’s age. Results: Clones C13-383 and C13-370 demonstrated a better performance than the controls, reaching higher crop yields than 195 t ha-1 of millable sugarcane, and 30 t Pol ha-1. The crop yields of C13-369 (164.48 t sugarcane t ha-1) and industrial yields (25.53 t Pol ha-1) were similar to cultivar C86-12 and higher than C1051-73. Conclusions: The three promising clones have a high genetic potential for sugar production, confirming its possible use as early-maturation cultivars to be harvested at the beginning of production

Phenological Characterization and Production of Biomass from 12 Varieties of Sugar to Feed Cattle

Context: One of the main limiting factors of cattle production in Cuba is linked to low availability of quality pasture in sufficient amounts, during the dry season. Sugar cane has anatomical and physiological features that offer advantages as food and energetic supplement to ruminants. Objective: To characterize 12 varieties of sugar cane for cattle nutrition. Methods: Phenological evaluations were made and biomass production was determined in a study conducted at the Territorial Station of Sugar Cane Research (ETICA), mid-east Camagüey, in dry lands. A randomized block experimental design was made, consisting of 12 treatments (varieties) and three replications. The phenological composition of the stump (stem, top, and whole) was determined at 14 months in all the varieties. Agronomic variables plant height, stem diameter, number of stems m-2, active leaves, and production of green biomass by fractions and as a whole, were determined as well. Results: The study demonstrated the existence of no significant differences in the phenological composition among the varieties. Concerning variables crop yield and green biomass production, varieties C92-325, C86-12, C99-374, C90-530, and C97-366 showed the greatest potential. Conclusions: Its use is recommended in the major cattle raising areas in the province and the country with similar edaphoclimatic conditions to the experimental area

Growth Indicators of a 48-Clone Sugar Cane Population (Saccharum spp.) with Forage Potential

The aim of this paper was to determine growth indicators in a 48-clone sugar cane population, with promising phenotypical features for forage production.   The following indicators were assessed: leaf area (A), leaf area index (LA1); leaf area ratio (LAR); specific leaf area (SLA); leaf weight ratio (LWR); crop growth rate (CGR); net assimilation rate (NAR); relative growth rate in weight (RGR); biomass production speed (G); leaf area duration (LAD); and biomass duration (Z), monthly (187 - 370 days). The minimum, the mean, the maximum values, and the population variance were determined for all cutting ages and the variables assessed. The results achieved have provided quantitative values that can be used as reference for selection and assessment of forage genotypes for ruminant nutrition

Tres nuevos clones promisorios de caña de azúcar de alta productividad agrícola e industrial

Context: At the present time it is necessary to have sugarcane cultivars with high agricultural and industrial potential to beginnings of harvest that allows to substitute those with many years of exploitation that have diminished their productivity and resistance to the main pest and diseases that affect to the sugarcane crop. Objective: To evaluate the agricultural and industrial potential of three new promissory clones of sugarcane. Methods: The experimental design was a random blocks with five treatments and three replications. The treatments were three promissory clones (C13-369, C13-370 and C13-383) and two commercial cultivars (C86-12 and C1051-73) used as control. Agrobotanic characteristics and the agricultural yield, as well as indicators of quality of the juices to harvest beginnings (December) at 12 months crop age in the plant cane crop were evaluated. Results: The C13-383 and C13-370 clones overcame the control when reaching agricultural yields superiors to 195 t ha-1 of millable cane and 30 t Pol ha-1. The C13-369 clone showed values of agricultural (164.48 t cane ha-1) and industrial (25.53 t Pol ha-1) yield similar when C86-12 cultivar and superiors to the C1051-73. Conclusions: The three promissory clones possess a high genetic potential for the sugar production and it reaffirms their possible use like cultivars of early maturity to be harvested to beginnings harvest period.Contexto: En la actualidad es necesario contar con cultivares de caña de azúcar con alto potencial agrícola e industrial a inicios de zafra que permita sustituir aquellos que con muchos años de explotación han disminuido su productividad y resistencia a las principales plagas y enfermedades que afectan al cultivo. Objetivo: Evaluar el potencial agroindustrial de tres nuevos clones promisorios de caña de azúcar en condiciones de secano. Métodos: Se utilizó un diseño experimental de bloques al azar con cinco tratamientos y tres réplicas. Los tratamientos fueron tres clones promisorios (C13-369, C13-370 y C13-383) y dos cultivares comerciales (C86-12 y C1051-73) utilizados como testigos. Se evaluaron características agrobotánicas y el rendimiento agrícola, así como indicadores de calidad de los jugos a inicios de zafra (diciembre) con 12 meses de edad el cultivo en la cepa de caña planta. Resultados: Los clones C13-383 y C13-370 superaron a los testigos al alcanzar rendimientos agrícolas superiores a las 195 t ha-1 de caña molible y 30 t Pol ha-1. El clon C13-369 mostró valores de rendimiento agrícola (164,48 t caña ha-1) e industrial (25,53 t Pol ha-1) similares al cultivar C86-12 y superiores al C1051-73. Conclusiones: Los tres clones promisorios poseen un alto potencial genético para la producción de azúcar y reafirma su posible utilización como cultivares de madurez temprana para ser cosechados a inicios de zafra

Tres nuevos clones promisorios de caña de azúcar de alta productividad agrícola e industrial

Contexto: En la actualidad es necesario contar con cultivares de caña de azúcar con alto potencial agrícola e industrial a inicios de zafra que permita sustituir aquellos que con muchos años de explotación han disminuido su productividad y resistencia a las principales plagas y enfermedades que afectan al cultivo. Objetivo: Evaluar el potencial agroindustrial de tres nuevos clones promisorios de caña de azúcar en condiciones de secano. Métodos: Se utilizó un diseño experimental de bloques al azar con cinco tratamientos y tres réplicas. Los tratamientos fueron tres clones promisorios (C13-369, C13-370 y C13-383) y dos cultivares comerciales (C86-12 y C1051-73) utilizados como testigos. Se evaluaron características agrobotánicas y el rendimiento agrícola, así como indicadores de calidad de los jugos a inicios de zafra (diciembre) con 12 meses de edad el cultivo en la cepa de caña planta. Resultados: Los clones C13-383 y C13-370 superaron a los testigos al alcanzar rendimientos agrícolas superiores a las 195 t ha-1 de caña molible y 30 t Pol ha-1. El clon C13-369 mostró valores de rendimiento agrícola (164,48 t caña ha-1) e industrial (25,53 t Pol ha-1) similares al cultivar C86-12 y superiores al C1051-73. Conclusiones: Los tres clones promisorios poseen un alto potencial genético para la producción de azúcar y reafirma su posible utilización como cultivares de madurez temprana para ser cosechados a inicios de zafra

Caracterización fenológica y producción de biomasa de 12 variedades de caña de azúcar para la alimentación bovina

Context: One of the main limitations of bovine production in Cuba is associated with the low availability of pastures in quantity and quality in the dry season of the year. Sugarcane has anatomical and physiological characteristics that offer advantages as food and energy supplement for ruminants. Objective:  In order to characterize 12 sugarcane varieties for bovine feeding. Methods: Phenological evaluations and biomass production determination were carried out in a study planted in areas of the Sugarcane Research Territorial Station of Camagüey under rainfed conditions. An experimental design of random blocks with 12 treatments (varieties) and three replicas was used. The phenological composition (stalk; sugarcane top and whole fraction) was determined at 14 months of age in the plant cane ratoon, as well as the agronomic variables height of the plant, diameter of the stalk, number of stalk m-2, active leaves and green biomass production for fractions of the plant and in whole way. Results: It was shown that there are not significant differences in the phenological composition among sugarcane varieties. With regard to agricultural yield variables and green biomass production the C92-325, C86-12, C99-374, C90-530 and C97-366 varieties expressed the highest potentials. Conclusions: It is recommended the use of these varieties in the main cattle areas of the province and the country that have similar soil and climate conditions to the area where this study was carried out.Contexto: Una de las principales limitantes de la producción bovina en Cuba está asociada a la baja disponibilidad de pastos en cantidad y calidad en el período poco lluvioso del año. La caña de azúcar posee características anatómicas y fisiológicas que ofrecen ventajas como alimento y suplemento energético de rumiantes. Objetivo: Con el objetivo de caracterizar 12 variedades de caña de azúcar para la alimentación bovina. Métodos: Se realizaron evaluaciones fenológicas y determinación de producción de biomasa en un estudio plantado en áreas de la Estación Territorial de Investigaciones de la Caña de Azúcar (ETICA) Centro Oriental de Camagüey en condiciones de secano. Se utilizó un diseño experimental de bloques al azar con 12 tratamientos (variedades) y tres réplicas. A los 14 meses de edad en la cepa de caña planta se determinó la composición fenológica (tallo, cogollo e integral) por variedad, así como las variables agronómicas altura de la planta, diámetro del tallo, número de tallos m-2, hojas activas y producción de biomasa verde por fracciones de la planta y de forma integral. Resultados: Se demostró que no existen diferencias significativas en la composición fenológica entre variedades. Con respecto a las variables del rendimiento agrícola y la producción de biomasa verde las variedades C92-325, C86-12, C99-374, C90-530 y C97-366 expresaron los mayores potenciales. Conclusiones: Se recomienda su uso en las principales áreas ganaderas de la provincia y el país que posean similares condiciones edafoclimáticas a la zona donde se llevó a cabo este estudio