Functional Impact and Regulation of Alternative Splicing in Mouse Heart Development and Disease.

Alternative splicing (AS) plays a major role in the generation of transcript diversity. In the heart, roles have been described for some AS variants, but the global impact and regulation of AS patterns are poorly understood. Here, we studied the AS profiles in heart disease, their relationship with heart development, and the regulatory mechanisms controlling AS dynamics in the mouse heart. We found that AS profiles characterized the different groups and that AS and gene expression changes affected independent genes and biological functions. Moreover, AS changes, specifically in heart disease, were associated with potential protein-protein interaction changes. While developmental transitions were mainly driven by the upregulation of MBNL1, AS changes in disease were driven by a complex regulatory network, where PTBP1 played a central role. Indeed, PTBP1 over-expression was sufficient to induce cardiac hypertrophy and diastolic dysfunction, potentially by perturbing AS patterns.S

Lafora Disease Is an Inherited Metabolic Cardiomyopathy

This work was supported by grants from the Spanish Ministry of Economy and Competitiveness (SAF2015-65722-R to Dr. Lara-Pezzi and SAF2014-59594-R to Dr. Serratosa), Autonomous Community of Madrid (2010-BMD2321, FIBROTEAM Consortium), European Union's FP7 (CardioNeT-ITN-289600, CardioNext-ITN-608027), the Spanish Carlos III Institute of Health (CPII14/00027 to Dr. Lara-Pezzi, PI13/00865 to Dr. Sanchez and RD12/0042/066 to Drs. Garcia-Pavia and Lara-Pezzi), and the National Institute of Neurological Disorders And Stroke of the National Institutes of Health (P01NS097197 to Dr. Sanchez). This work was also supported by the Plan Estatal de I+D+I 2013-2016-European Regional Development Fund (FEDER) "A way of making Europe," Spain. The Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC) is supported by the Spanish Ministry of Economy and Competitiveness (MINECO) and the Pro-CNIC Foundation, and is a Severo Ochoa Center of Excellence (MINECO award SEV-2015-0505).S

Lung ultrasound as a translational approach for non-invasive assessment of heart failure with reduced or preserved ejection fraction in mice

Aims: Heart failure (HF) has become an epidemic and constitutes a major medical, social, and economic problem worldwide. Despite advances in medical treatment, HF prognosis remains poor. The development of efficient therapies is hampered by the lack of appropriate animal models in which HF can be reliably determined, particularly in mice. The development of HF in mice is often assumed based on the presence of cardiac dysfunction, but HF itself is seldom proved. Lung ultrasound (LUS) has become a helpful tool for lung congestion assessment in patients at all stages of HF. We aimed to apply this non-invasive imaging tool to evaluate HF in mouse models of both systolic and diastolic dysfunction. Methods and results: We used LUS to study HF in a mouse model of systolic dysfunction, dilated cardiomyopathy, and in a mouse model of diastolic dysfunction, diabetic cardiomyopathy. LUS proved to be a reliable and reproducible tool to detect pulmonary congestion in mice. The combination of LUS and echocardiography allowed discriminating those mice that develop HF from those that do not, even in the presence of evident cardiac dysfunction. The study showed that LUS can be used to identify the onset of HF decompensation and to evaluate the efficacy of therapies for this syndrome. Conclusions: This novel approach in mouse models of cardiac disease enables for the first time to adequately diagnose HF non-invasively in mice with preserved or reduced ejection fraction, and will pave the way to a better understanding of HF and to the development of new therapeutic approaches.This study was supported by grants from the Spanish Ministerio de Economia y Competitividad (SAF2015-65722-R), Comunidad Autonoma de Madrid (2010-BMD2321, FIBROTEAM Consortium), European Union's FP7 (CardioNeT-ITN-289600, CardioNext-ITN-608027) and the Spanish Instituto de Salud Carlos III (CPII14/00027 to E.L-P, RD12/0042/0054 to B.I. and RD12/0042/066 to P.G.-P. and E.L-P). This work was also supported by the Plan Estatal de I+D+I 2013-2016 - European Regional Development Fund (FEDER) "A way of making Europe", Spain. The CNIC is supported by the Ministry of Economy, Industry and Competitiveness (MINECO) and the Pro CNIC Foundation, and is a Severo Ochoa Center of Excellence (MINECO award SEV-2015-0505).S

Activation of Serine One-Carbon Metabolism by Calcineurin A beta 1 Reduces Myocardial Hypertrophy and Improves Ventricular Function

BACKGROUND In response to pressure overload, the heart develops ventricular hypertrophy that progressively decompensates and leads to heart failure. This pathological hypertrophy is mediated, among others, by the phosphatase calcineurin and is characterized by metabolic changes that impair energy production by mitochondria. OBJECTIVES The authors aimed to determine the role of the calcineurin splicing variant CnA beta 1 in the context of cardiac hypertrophy and its mechanism of action. METHODS Transgenic mice overexpressing CnAb1 specifically in cardiomyocytes and mice lacking the unique C-terminal domain in CnA beta 1 (CnA beta 1(Delta i12) mice) were used. Pressure overload hypertrophy was induced by transaortic constriction. Cardiac function was measured by echocardiography. Mice were characterized using various molecular analyses. RESULTS In contrast to other calcineurin isoforms, the authors show here that cardiac-specific overexpression of CnA beta 1 in transgenic mice reduces cardiac hypertrophy and improves cardiac function. This effect is mediated by activation of serine and one-carbon metabolism, and the production of antioxidant mediators that prevent mitochondrial protein oxidation and preserve ATP production. The induction of enzymes involved in this metabolic pathway by CnAb1 is dependent on mTOR activity. Inhibition of serine and one-carbon metabolism blocks the beneficial effects of CnA beta 1. CnA beta 1(Delta i12) mice show increased cardiac hypertrophy and declined contractility. CONCLUSIONS The metabolic reprogramming induced by CnAb1 redefines the role of calcineurin in the heart and shows for the first time that activation of the serine and one-carbon pathway has beneficial effects on cardiac hypertrophy and function, paving the way for new therapeutic approaches. (J Am Coll Cardiol 2018; 71: 654-67) (C) 2018 The Authors. Published by Elsevier on behalf of the American College of Cardiology Foundation. This is an open access article under the CC BY-NC-ND license (http://creativecommons. org/licenses/by-nc-nd/4.0/).This work was supported by grants from the European Union (CardioNeT-ITN-289600 and CardioNext-608027 to Dr. Lara-Pezzi; Meet-ITN-317433 to Dr. Enriquez; UE0/MCA1108 to Dr. Acin-Perez), from the Spanish Ministry of Economy and Competitiveness (SAF2015-65722-R and SAF2012-31451 to Dr. Lara-Pezzi; SAF2015-71521-REDC, BFU2013-50448, and SAF2012-32776 to Dr. Enriquez; RyC-2011-07826 to Dr. Acin-Perez; BIO2012-37926 and BIO2015-67580-P to Dr. Vazquez), from the Spanish Carlos III Institute of Health (CPII14/00027 to Dr. Lara-Pezzi; RD12/0042/066 to Drs. Garcia-Pavia and Lara-Pezzi), from the Regional Government of Madrid (2010-BMD-2321 ``Fibroteam´´ to Dr. Lara-Pezzi; 2011-BMD-2402 ``Mitolab´´ to Dr. Enriquez) and the FIS-ISCIII (PRB2-IPT13/0001 and RD12/0042/0056-RIC-RETICS to Dr. Vazquez). This work was also supported by the Plan Estatal de IthornDthornI 2013-2016-European Regional Development Fund (FEDER) ``A way of making Europe,´´ Spain. The CNIC is supported by the Spanish Ministry of Economy and Competitiveness and by the Pro-CNIC Foundation and is a Severo Ochoa Center of Excellence (MINECO award SEV-2015-0505). Drs. Vazquez and Garcia-Pavia have served as consultants for VL39. All other authors have reported that they have no relationships relevant to the contents of this paper to disclose. Drs. Padron-Barthe, Villalba-Orero, and Gomez-Salinero contributed equally to this work and are joint first authors. Robyn Shaw, MD, PhD, served as Guest Editor for this paper.S

Measurement of vegetations in infective endocarditis: An inaccurate method to decide the therapeutical approach

Background: The European Society of Cardiology and American Heart Association guidelines give a central role to the maximal vegetation diameter in the indication for surgery to prevent embolism in left sided infective endocarditis (LSIE). Vegetation measuring is likely to be inaccurate. The hypothesis herein, is that the vegetation diameter is not an appropriate surgical criterion given the variability of its measurement. Methods: Two trained echocardiographers independently measured the maximal vegetation diameter by transesophageal echocardiogram of 76 vegetations in 67 consecutive patients with definite infective endocarditis in an off-line workstation. The interobserver variability was calculated by the interclass correlation coefficient. The relationship between the strength of agreement for the cut-off points of 10 and 15 mm was also calculated. Finally, the number of patients whose surgical indication would have changed depending on which operator measured the vegetation was evaluated. Results: Interobserver interclass correlation coefficient in the measurement of the maximal longitudinal diameter of the vegetations was 0.757 (0.642–0.839). The strength of agreement of the interobserver analysis for the cut-off point of 10 mm was 0.533 (0.327–0.759). For the cut-off point of 15 mm it was 0.475 (0.270–0.679). If heart failure or uncontrolled infections had been absent, the surgical indication would have changed in a total of 33 patients (33/76; 43%) depending on which operator measured the vegetation. Conclusions: The variability in the measurements of the maximal longitudinal diameter by transesophageal echocardiogram is high. Surgical indications based on the cut-off points recommended by the international guidelines should be revised

Efecto de las propiedades físicas, químicas, biológicas del suelo bajo siembra directa y trasplante en combinación con abonos verdes antecesores, sobre la producción de hortalizas en sistemas agroecológicos

Durante los años 2007-2012 fueron analizados dos tratamientos de sistemas de implantación de cultivos hortícolas; uno siembra directa de semillas (SD) y otro transplante de plantines (TR). Estos se combinaron con cuatro antecesores de manejo de suelos. Los tratamientos antecesores fueron: (H1) barbecho desnudo estival antes del policultivo de lechuga, acelga, remolacha, (H2) moha antes del policultivo de lechuga, acelga, remolacha, (H3) sorgo forrajero antes del cultivo de cebolla de verdeo, (H4) avena antes del cultivo de zapallo. El experimento tuvo diseño factorial con dos sistema de implantación, cuatro antecesores y tres repeticiones. En el período 2007-2009 fueron analizadas las variables, físicas, químicas, biológicas de la matriz del suelo: densidad aparente (DAP), infiltración básica (IB), porosidad total (PT), distribución de macroporos (MP), mesoporos (MSP), microporos (MCP), estabilidad de agregados (IEA), pH, conductividad eléctrica (CE), sulfatos (SO4), carbono orgánico total (COT), nitrógeno total (NT), fósforo extractable (Pe), calcio (Ca), magnesio (Mg), potasio (K), sodio (Na), capacidad de intercambio catiónico (CIC), respiración (AB), carbono en la biomasa microbiana (CBM), coeficientes metabólicos (qCO2), fluoresceína difosfato (FDA) y fosfatasa ácida (Pasa). También fueron registrados los rendimientos de las hortalizas luego de los antecesores. El análisis estadístico de todas las variables se realizó mediante dos procedimientos, modelos lineales (ANOVA, STEPWISE) y componentes principales (ACP). Existieron interacciones significativas entre sistema de implantación SD y TR y antecesores H1, H2, H3, H4, pero no fueron tan acentuadas, como la separación de tratamientos de SD y TR mediante ACP, lo que explicó el 83,4% de la variación ocurrida. Las variables biológicas y la IB explicaron en mayor medida los resultados obtenidos al favor del TR en los rendimientos de lechuga y acelga. Por el contrario las variables químicas y el IEA explicaron en mayor medida los resultados obtenidos a favor de la SD en el rendimiento de remolacha.EEA San PedroFil: Ullé, Jorge Angel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria San Pedro; ArgentinaFil: Faggioli, Valeria Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juarez; ArgentinaFil: Serri, Dannae Lilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; ArgentinaFil: Ortega y Villasana, Pilar. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación Investigación y Desarrollo Tecnológico para la Agricultura Familiar Región NEA; ArgentinaFil: Darder, María Liliana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Dalpiaz, María Jimena. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: García, Leticia Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Farroni, Abel Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Rimatori, Fernando Miguel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Colombini, F. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Villalba, D. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; Argentin

Indicadores de calidad edáfica en cultivos antecesores consociados y monocultivo de batata

Un experimento con batata cv Arapey, fue implantado en 2011 en Argiudoles evaluando diferentes tipos de manejo de suelos. Fueron comparados los manejos de mayor adopción como monocultivo y antecesor barbecho desnudo, con otras prácticas de manejo conservacionistas como antecesores de variedades de maíz (cv. Blanco Duro, cv. Caiano, cv Azteca), sorgo forrajero (cv. Talero), leguminosas subtropicales (Canavalia ensiformis, Mucuna cinza) y cultivo de batata (cv Arapey) de forma consociada en hileras con estos (cv Blanco Duro, Cv Caiano, Canavalia ensiformis). Fueron implementados 12 tratamientos y tres repeticiones, en diseño en bloques aleatorizados (DBCA) en un ensayo con objetivo de larga duración en la EEA San Pedro. Las batatas fueron trasplantadas, en 2011, 2012, 2013, siendo aquí comunicados los resultados de las primeras dos cosechas. En el primer año se cosecharon y registraron los rendimientos (ha) de los tratamientos de batata monocultivo, batata consociada con variedades maíz (cv Blanco Duro, cv. Caiano) y batata con leguminosas (Canavalia ensiformis). En el segundo año se cosecharon los tratamientos luego de los antecesores, barbecho desnudo, maíz (cv. Blanco Duro, cv. Caiano, cv Azteca), sorgo (cv. Talero) y leguminosas (Canavalia ensiformis). En cada situación fueron levantadas para la determinación las variables edáficas, físicas, químicas, biológicas: densidad aparente (DAP), infiltración básica (IB), porosidad total (PT), distribución de macroporos (MP), mesoporos (MSP), microporos (MCP), estabilidad de agregados (EA), pH, conductividad eléctrica (CE), carbono orgánico total (COT), nitrógeno total (NT), fósforo extractable (Pe), calcio (Ca), magnesio (Mg), potasio (K), sodio (Na), capacidad de intercambio catiónico (CIC), respiración (AB), carbono en la biomasa microbiana (CBM), coeficientes metabólicos (qCO2), fluoresceína difosfato (FDA) , fosfatasa ácida (Pasa), glomalinas (PROT) e hifas (H) arbúsculos (A) y vesículas (V) de micorrizas. El objetivo del trabajo fue conocer en qué medida el conjunto de variables-indicadores, permitían diferenciar las situaciones de manejo. En el análisis ANOVA, en el primer año 2012 los rendimientos de batata no difirieron significativamente, mientras que en el segundo 2013, la batata monocultivo, el antecesor barbecho desnudo, sorgo (cv. Talero) y leguminosa (Canavalia ensiformis), superaron significativamente a las batatas consociadas en hileras con variedades de maíz (cv Blanco Duro, Cv Caiano) y Canavalia ensiformis. El análisis de regresión múltiple (stepwise) en el primer año 2012, demostró un alta relación del rendimiento de batata con las variables biológicas, (CBM), (PROT), (qCO2) (R²= 0.65) y las físicas (MP), (MCP) (R²= 0.66). En el segundo año 2013, los rendimientos de batata presentaron una baja relación en sentido negativa con las variables biológicas (FDA) (Pasa), (PROT) (R²= 0.40), y las físicas (EA) (MSP) (R²= 0.26), pero una alta relación positiva con el Nt (R²= 0.85). El análisis de componentes principales separó mediante el CP1 (38,7% de la variación ocurrida), los tratamientos consociados del monocultivo, demostrando los consociados agruparse en un cuadrante junto a los rendimientos de batata 2012, la (FDA), (Pasa), (PROT), (MCP), (EA). Por el contrario, los de monocultivo se agruparon junto a los rendimientos de batata 2013, las variables químicas en su totalidad, las físicas (MP), (MSP) y micorrizas (H), (A), (V) en otro extremo.EEA San PedroFil: Ullé, Jorge Angel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria San Pedro; ArgentinaFil: Marti, Héctor Rubén.Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria San Pedro; ArgentinaFil: Faggioli, Valeria Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juarez; ArgentinaFil: Darder, María Liliana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Dalpiaz, María Jimena. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Farroni, Abel Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Rimatori Fernando Miguel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Colombini, F. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Villalba, D. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Martinez, Fortunato. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación Investigación y Desarrollo Tecnológico para la Agricultura Familiar Región NEA; ArgentinaFil: Ortega y Villasana, Pilar. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación Investigación y Desarrollo Tecnológico para la Agricultura Familiar Región NEA; Argentin

Improving the reporting quality of intervention trials addressing the inter-individual variability in response to the consumption of plant bioactives: quality index and recommendations

PURPOSE: The quality of the study design and data reporting in human trials dealing with the inter-individual variability in response to the consumption of plant bioactives is, in general, low. There is a lack of recommendations supporting the scientific community on this topic. This study aimed at developing a quality index to assist the assessment of the reporting quality of intervention trials addressing the inter-individual variability in response to plant bioactive consumption. Recommendations for better designing and reporting studies were discussed. METHODS: The selection of the parameters used for the development of the quality index was carried out in agreement with the scientific community through a survey. Parameters were defined, grouped into categories, and scored for different quality levels. The applicability of the scoring system was tested in terms of consistency and effort, and its validity was assessed by comparison with a simultaneous evaluation by experts' criteria. RESULTS: The "POSITIVe quality index" included 11 reporting criteria grouped into four categories (Statistics, Reporting, Data presentation, and Individual data availability). It was supported by detailed definitions and guidance for their scoring. The quality index score was tested, and the index demonstrated to be valid, reliable, and responsive. CONCLUSIONS: The evaluation of the reporting quality of studies addressing inter-individual variability in response to plant bioactives highlighted the aspects requiring major improvements. Specific tools and recommendations favoring a complete and transparent reporting on inter-individual variability have been provided to support the scientific community on this field

Efecto de las propiedades físicas, químicas, biológicas del suelo y su relación con el rendimiento de variedades de maíz y batata consociadas o en monocultivo

Durante 2011 y 2012 un experimento de variedades de maíz y otro de batata fueron llevados a cabo en monocultivo o en forma consociada en la EEA INTA San Pedro. Las variedades de maíz utilizadas fueron Caiano, Blanco Duro, Azteca, Chala Roja, Mato Grosso y la batata cv Arapey. En el experimento de maíz los tratamientos fueron en total 12, es decir: 5 variedades de maíz en monocultivo, 2 variedades de maíz consociadas con leguminosas, 2 variedades de leguminosas monocultivo (Canavalia ensiformis) (Mucuna cinereum), 1 cv Sorgo talero, 1 barbecho desnudo y 1 campo natural. En el experimento de batata los tratamientos fueron 12: 3 variedades de maíz monocultivo, 2 variedades de maíz consociadas con batata, 1 de batata consociada con leguminosas; 2 variedades de leguminosas monocultivo (Canavalia ensiformis) (Mucuna cinereum), 1 cv Sorgo talero; 1 de batata seguida de avena, 1 de batata en monocultivo y 1 barbecho desnudo. Fueron analizadas las variables, físicas, químicas, biológicas de suelo: densidad aparente (DAP), infiltración básica (IB), porosidad total (PT), distribución de macroporos (MP), mesoporos (MSP), microporos (MCP), estabilidad de agregados (IEA), pH, conductividad eléctrica (CE), sulfatos (SO4), carbono orgánico total (COT), nitrógeno total (NT), fósforo extractable (Pe), calcio (Ca), magnesio (Mg), potasio (K), sodio (Na), capacidad de intercambio catiónico (CIC), respiración (AB), carbono en la biomasa microbiana (CBM), coeficientes metabólicos (qCO2), fluoresceína difosfato (FDA) y fosfatasa ácida (Pasa), Glomalinas (PROT) y la macrofauna del suelos (Edaf). Resultados promisorios indican una alta perfomance de rendimientos en los tratamientos de M Caiano consociado a Canavalia ensiformis y M Caiano consociado a batata, igualando al monocultivo de ambos. En el experimento de batata fue detectada un R² Aj = 0.66 seleccionando las variables MP, MCP, qCO2, PROT. En el experimento maíz fue observado R² Aj = 0.38 seleccionando CBM; Pasa, PROT y Edaf.EEA San PedroFil: Ullé, Jorge Angel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria San Pedro; ArgentinaFil: Faggioli, Valeria Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juarez; ArgentinaFil: Martí, Héctor Rubén. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria San Pedro; ArgentinaFil: Serri, Dannae Lilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; ArgentinaFil: Ortega y Villasana, Pilar. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación Investigación y Desarrollo Tecnológico para la Agricultura Familiar Región NEA; ArgentinaFil: Dalpiaz, María Jimena. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: García, Leticia Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Darder, María Liliana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Farroni, Abel Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Rimatori, Fernando Miguel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Colombini, Diego Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Villalba, Fabio Leonardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; Argentin