Efecto de la humedad del grano y el agregado de urea sobre la conservación alcalina del sorgo

La conservación alcalina de granos húmedos mediante el agregado de urea no requiere de anaerobiosis. El objetivo del presente trabajo fue evaluar los efectos del agregado de distintas proporciones de urea sobre la conservación del grano de sorgo con distintos contenidos de humedad. El ensayo tuvo lugar en la EEA Anguil "Ing. Agr. Guillermo Covas" del INTA. Se definieron los factores: Humedad: 16, 18, 22,26 Y30% de humedad en el grano; Urea: 1, 2 y 3 % de urea (base seca). Se establecieron 4 repeticiones por combinación de factores (15 tratamientos). Cada repetición consistió en 20 de grano de sorgo (en base seca) con humedad reconstituida hasta los niveles deseados, a las que se incorporaron los niveles de urea. Se utilizaron bolsas de nylon permitiendo un acceso limitado al aire. Para mantener la aerobiosis, el contenido de las bolsas se mezcló a mano semanalmente durante la duración del ensayo. A partir del día O (momento del agregado de la urea) y en los días 1,4, 7, 18,30 Y90 de iniciado el ensayo se determinó desarrollo de biomasa fúngica, temperatura y pH. En los días O y 90 se muestreo para determinar el peso de mil granos, y los contenidos de proteína bruta (pB; N-Kjeldahl * 6,25) y fibra detergente ácido (FDA). Entre ambos factores, la humedad del grano tuvo un efecto significativo (p < 0,001) Y no así el nivel de urea agregada (P = 0,549). Los tratamientos con 22% de humedad o más y 1% de urea registraron un moderado desarrollo de hongos a partir de los 18 días de agregada la urea. En los tratamientos que incluyeron 2 o 3% de urea no se registró desarrollo visible de hongos. Los tratamientos con 1% de urea y 26 o 30% de humedad mostraron un mayor contenido de FDA (P < 0,05), comparados con su situación inicial y con los otros tratamientos. Se evidenció un efecto lineal (P < 0.05) entre la pérdida de peso y el incremento de la humedad del grano almacenado a partir del 18% de humedad combinado con 1% de urea. Esta experiencia indica que un agregado de 2% de urea (sobre base seca) sería necesario para lograr una conservación de granos con 18% o más de humedad.Director: Ing. Agr. Anibal Pordomingo. Cátedra Zootécnia I

Oral Delivery of the P2Y12 Receptor Antagonist Ticagrelor Prevents Loss of Photoreceptors in an ABCA4−/− Mouse Model of Retinal Degeneration

PURPOSE. Accumulation of lysosomal waste is linked to neurodegeneration in multiple diseases, and pharmacologic enhancement of lysosomal activity is hypothesized to reduce pathology. An excessive accumulation of lysosomal-associated lipofuscin waste and an elevated lysosomal pH occur in retinal pigment epithelial cells of the ABCA4 mouse model of Stargardt\u27s retinal degeneration. As treatment with the P2Y12 receptor antagonist ticagrelor was previously shown to lower lysosomal pH and lipofuscin-like autofluorescence in these cells, we asked whether oral delivery of ticagrelor also prevented photoreceptor loss. METHODS. Moderate light exposure was used to accelerate photoreceptor loss in albino ABCA4 mice as compared to BALB/c controls. Ticagrelor (0.1%–0.15%) was added to mouse chow for between 1 and 10 months. Photoreceptor function was determined with electroretinograms, while cell survival was determined using optical coherence tomography and histology. RESULTS. Protection by ticagrelor was demonstrated functionally by using the electroretinogram, as ticagrelor-treated ABCA4 mice had increased a-and b-waves compared to untreated mice. Mice receiving ticagrelor treatment had a thicker outer nuclear layer, as measured with both optical coherence tomography and histologic sections. Ticagrelor decreased expression of LAMP1, implicating enhanced lysosomal function. No signs of retinal bleeding were observed after prolonged treatment with ticagrelor. CONCLUSIONS. Oral treatment with ticagrelor protected photoreceptors in the ABCA4 mouse, which is consistent with enhanced lysosomal function. As mouse ticagrelor exposure levels were clinically relevant, the drug may be of benefit in preventing the loss of photoreceptors in Stargardt’s disease and other neurodegenerations associated with lysosomal dysfunction. © 2019 The Authors. All rights reserved

Polarized Cytokine Release Triggered by P2X7 Receptor from Retinal Pigmented Epithelial Cells Dependent on Calcium Influx

Cytokine release from non-inflammatory cells is a key step in innate immunity, and agonists triggering cytokine release are central in coordinating responses. P2X7 receptor (P2X7R) stimulation by extracellular ATP is best known to active the NLRP3 inflammasome and release IL-1β, but stimulation also leads to release of other cytokines. As cytokine signaling by retinal pigmented epithelial (RPE) cells is implicated in retinal neurodegeneration, the role of P2X7R in release of cytokine IL-6 from RPE cells was investigated. P2X7R stimulation triggered IL-6 release from primary mouse RPE, human iPS-RPE and human ARPE-19 cells. IL-6 release was polarized, with predominant rise across apical membranes. IL-6 release was inhibited by P2X7R antagonists A438079, A839977, and AZ10606120, but not the NRTI lamivudine (3TC), P2X1R antagonist NF279, or P2Y1R antagonist MRS2179. P2X7R-mediated IL-6 release required extracellular Ca2+ and was blocked by Ca2+ chelator BAPTA. IL-6 release and Ca2+ elevation occurred rapidly, consistent with vesicular IL-6 staining in unstimulated cells. P2X7R stimulation did not trigger IL-1β release in these unprimed cells. P2X7R-mediated IL-6 release was enhanced in RPE cells from the ABCA4-/- mouse model of retinal degeneration. In summary, P2X7R stimulation triggers rapid Ca2+-dependent IL-6 release across the apical membrane of RPE cells

Relación entre el valor del ratio elastográfico y la clasificación citológica de Bethesda en la patología tiroidea

ResumenObjetivoPresentar nuestra experiencia en la categorización de la patología tiroidea, a través de la utilización de parámetros ecográficos de malignidad y elastografía con medición del ratio de la deformación tisular, y la correlación de los hallazgos obtenidos con la clasificación citológica de Bethesda.Materiales y métodosSe llevó a cabo un estudio prospectivo y observacional, entre septiembre de 2012 y abril de 2013, que incluyó 137 nódulos tiroideos. Se excluyeron 10 casos Bethesda III-IV. Se realizó ecografía, power Doppler, visualización de micropartículas (Micropure) y elastografía con medición del ratio elastográfico, así como también punción aspirativa con aguja fina guiada por ecografía (con el citólogo presente), utilizando la clasificación Bethesda. Los estudios fueron hechos por el mismo operador con un ecógrafo Toshiba Aplio 400 y los datos estadísticos se evaluaron con el programa IBM SPSS Statistics 20.ResultadosSe estudiaron 127 nódulos en pacientes con una edad promedio de 59±16 años. El 82% de los casos ocurrió en mujeres. Ciento veinte nódulos (94%) fueron clasificados como Bethesda II. La media elastográfica para Bethesda I-II fue de 1,94±2,12 vs. 7,07±5,46 para V-VI (p: 0,048). El punto de corte elastográfico ≤ 2 (87 de 127) presentó una sensibilidad del 85,7% y una especificidad del 81,7% para predecir Bethesda asociada a patología benigna, con un valor predictivo negativo (VPN) del 99% y un valor predictivo positivo del 15%.ConclusionesEl ratio elastográfico permitió descartar la patología tiroidea maligna con valores ≤ 2 y un VPN del 99%, mejorando la selección de los pacientes a punzar. El incremento del ratio elastográfico se asoció a una mayor probabilidad de patología maligna, aunque no se pudo establecer un valor de corte debido al bajo número de casos con Bethesda V-VI.AbstractObjectivesWe present our experience in the categorization of thyroid pathology using the sonographic parameters of malignancy and elastography with measurement elastography strain ratio, to evaluate the relationship between the results found and the Bethesda classification.Materials and methodsProspective observational study, included 137 thyroid nodules studied between September 2012- April 2013. We excluded 10 cases with Bethesda categories III-IV. Ultrasonography, Doppler, Micropure, elastogrphy strain ratio between the lesion and the normal tissue, fine needle aspiration cytology (FNAC),were the diagnosis methods used. The pathologist was always present and the cytological classi fication of Bethesda was used. All study was made by the same physician used Toshiba Aplio 400 ultrasound unit. Results were analyzed with IBM SPSS Statistics 20.ResultsWe studied 127 nodules in patients 59±16 years old, 82% were female; 120 were Bethesda II (94%). The average strain ratio for nodules Bethesda I-II was 1.94±2.12 vs. 7.07±5.46 for those nodules Bethesda V-VI (p:0,048). This means that an elastography strain ratio ≤ 2 (87 of 127 nodules) has a sensibility of 85.7% and a specificity of 81.7% of predicting Bethesda associated with benign pathology with a negative predictive value (NPV) of 99% and a positive predictive value of 15%.ConclusionThe elastography strain ratio allowed to discard malignant nodules with strain ratio ≤ 2 with a NPV of 99% improves the selection of patients for FNAC. The increment in the elastography strain ratio was associated to a higher possibility of malignant thyroid pathology, being unable to determine a limit value due to the low amount of cases with nodules Bethesda V-VI

Stimulation of TLR3 Triggers Release of Lysosomal ATP in Astrocytes and Epithelial Cells that Requires TRPML1 Channels

Cross-reactions between innate immunity, lysosomal function, and purinergic pathways may link signaling systems in cellular pathologies. We found activation of toll-like receptor 3 (TLR3) triggers lysosomal ATP release from both astrocytes and retinal pigmented epithelial (RPE) cells. ATP efflux was accompanied by lysosomal acid phosphatase and beta hexosaminidase release. Poly(I:C) alkalinized lysosomes, and lysosomal alkalization with bafilomycin or chloroquine triggered ATP release. Lysosomal rupture with glycyl-L-phenylalanine-2-naphthylamide (GPN) eliminated both ATP and acid phosphatase release. Secretory lysosome marker LAMP3 colocalized with VNUT, while MANT-ATP colocalized with LysoTracker. Unmodified membrane-impermeant 21-nt and non-targeting scrambled 21-nt siRNA triggered ATP and acid phosphatase release, while smaller 16-nt RNA was ineffective. Poly(I:C)-dependent ATP release was reduced by TBK-1 block and in TRPML1-/- cells, while TRPML activation with ML-SA1 was sufficient to release both ATP and acid phosphatase. The ability of poly(I:C) to raise cytoplasmic Ca2+ was abolished by removing extracellular ATP with apyrase, suggesting ATP release by poly(I:C) increased cellular signaling. Starvation but not rapamycin prevented lysosomal ATP release. In summary, stimulation of TLR3 triggers lysosomal alkalization and release of lysosomal ATP through activation of TRPML1; this links innate immunity to purinergic signaling via lysosomal physiology, and suggests even scrambled siRNA can influence these pathways. © 2018 The Author(s)

Stimulation of TLR3 Triggers Release of Lysosomal ATP in Astrocytes and Epithelial cells that Requires TRPML1 Channels

Cross-reactions between innate immunity, lysosomal function, and purinergic pathways may link signaling systems in cellular pathologies. We found activation of toll-like receptor 3 (TLR3) triggers lysosomal ATP release from both astrocytes and retinal pigmented epithelial (RPE) cells. ATP efflux was accompanied by lysosomal acid phosphatase and beta hexosaminidase release. Poly(I:C) alkalinized lysosomes, and lysosomal alkalization with bafilomycin or chloroquine triggered ATP release. Lysosomal rupture with glycyl-L-phenylalanine-2-naphthylamide (GPN) eliminated both ATP and acid phosphatase release. Secretory lysosome marker LAMP3 colocalized with VNUT, while MANT-ATP colocalized with LysoTracker. Unmodified membrane-impermeant 21-nt and “non-targeting” scrambled 21-nt siRNA triggered ATP and acid phosphatase release, while smaller 16-nt RNA was ineffective. Poly(I:C)-dependent ATP release was reduced by TBK-1 block and in TRPML1−/− cells, while TRPML activation with ML-SA1 was sufficient to release both ATP and acid phosphatase. The ability of poly(I:C) to raise cytoplasmic Ca2+ was abolished by removing extracellular ATP with apyrase, suggesting ATP release by poly(I:C) increased cellular signaling. Starvation but not rapamycin prevented lysosomal ATP release. In summary, stimulation of TLR3 triggers lysosomal alkalization and release of lysosomal ATP through activation of TRPML1; this links innate immunity to purinergic signaling via lysosomal physiology, and suggests even scrambled siRNA can influence these pathways

The P2Y12 Receptor Antagonist Ticagrelor Reduces Lysosomal pH and Autofluorescence in Retinal Pigmented Epithelial Cells From the ABCA4-/- Mouse Model of Retinal Degeneration

The accumulation of partially degraded lipid waste in lysosomal-related organelles may contribute to pathology in many aging diseases. The presence of these lipofuscin granules is particularly evident in the autofluorescent lysosome-associated organelles of the retinal pigmented epithelial (RPE) cells, and may be related to early stages of age-related macular degeneration. While lysosomal enzymes degrade material optimally at acidic pH levels, lysosomal pH is elevated in RPE cells from the ABCA4-/- mouse model of Stargardt\u27s disease, an early onset retinal degeneration. Lowering lysosomal pH through cAMP-dependent pathways decreases accumulation of autofluorescent material in RPE cells in vitro, but identification of an appropriate receptor is crucial for manipulating this pathway in vivo. As the P2Y12 receptor for ADP is coupled to the inhibitory Gi protein, we asked whether blocking the P2Y12 receptor with ticagrelor could restore lysosomal acidity and reduce autofluorescence in compromised RPE cells from ABCA4-/- mice. Oral delivery of ticagrelor giving rise to clinically relevant exposure lowered lysosomal pH in these RPE cells. Ticagrelor also partially reduced autofluorescence in the RPE cells of ABCA4-/- mice. In vitro studies in ARPE-19 cells using more specific antagonists AR-C69931 and AR-C66096 confirmed the importance of the P2Y12 receptor for lowering lysosomal pH and reducing autofluorescence. These observations identify P2Y12 receptor blockade as a potential target to lower lysosomal pH and clear lysosomal waste in RPE cells. © 2018 Lu, Gómez, Lim, Guha, O\u27Brien-Jenkins, Coffey, Campagno, McCaughey, Laties, Carlsson and Mitchell

Polarized Cytokine Release Triggered by P2X7 Receptor from Retinal Pigmented Epithelial Cells Dependent on Calcium Influx

Cytokine release from non-inflammatory cells is a key step in innate immunity, and agonists triggering cytokine release are central in coordinating responses. P2X7 receptor (P2X7R) stimulation by extracellular ATP is best known to active the NLRP3 inflammasome and release IL-1β, but stimulation also leads to release of other cytokines. As cytokine signaling by retinal pigmented epithelial (RPE) cells is implicated in retinal neurodegeneration, the role of P2X7R in release of cytokine IL-6 from RPE cells was investigated. P2X7R stimulation triggered IL-6 release from primary mouse RPE, human iPS-RPE and human ARPE-19 cells. IL-6 release was polarized, with predominant rise across apical membranes. IL-6 release was inhibited by P2X7R antagonists A438079, A839977, and AZ10606120, but not the NRTI lamivudine (3TC), P2X1R antagonist NF279, or P2Y1R antagonist MRS2179. P2X7R-mediated IL-6 release required extracellular Ca2+ and was blocked by Ca2+ chelator BAPTA. IL-6 release and Ca2+ elevation occurred rapidly, consistent with vesicular IL-6 staining in unstimulated cells. P2X7R stimulation did not trigger IL-1β release in these unprimed cells. P2X7R-mediated IL-6 release was enhanced in RPE cells from the ABCA4−/− mouse model of retinal degeneration. In summary, P2X7R stimulation triggers rapid Ca2+-dependent IL-6 release across the apical membrane of RPE cell

Tel Erani, Israel: Report of the 2018 Season and Its Background

En el mes de julio de 2018, un equipo de investigadores del Instituto de Historia Antigua Oriental “Dr. Abraham Rosenvasser” en el marco del Proyecto PICT-Raíces 2015-2943, participó en las excavaciones de la campaña arqueológica realizada en Tel Erani, Israel. Este sitio, excavado desde la década del ‘50 del siglo pasado, es clave para comprender las relaciones entre el sur de Palestina (Levante meridional) y Egipto durante la Edad del Bronce Antiguo IB (segunda mitad del IV° milenio a.C.), pues se ha hallado una presencia significativa de cultura material egipcia, incluyendo un tiesto con un serekh del rey Narmer, así como también cerámica de un estilo local característico, llamada Erani C, que ha podido ser identificada en el delta del Nilo (Tell el-Farkha) y en tumbas protodinásticas, como la denominada U-j de Abidos. Parte de estos hallazgos se relaciona con la presencia de por lo menos dos murallas superpuestas que posiblemente rodeaban la totalidad del tel, de aproximadamente 25 ha, por lo que se trataría de uno de los asentamientos fortificados más tempranos de Palestina. En la campaña del año 2018 se excavaron dos áreas: el Área D3, donde abunda el material egipcio junto con elementos locales, y el Área P-Q, correspondiente a una de las zonas donde se encuentran las fortificaciones. Los resultados de esta última campaña indican que estas murallas serían anteriores a la fase egipcia, es decir el Bronce Antiguo IB1, pero luego, durante el Bronce Antiguo IB2, las relaciones entre ambas regiones se habrían intensificado, con la posibilidad de que los egipcios hayan pasado a tener un rol más activo en Tel Erani.In July 2018, a team of researchers from the Institute of Ancient Near Eastern History “Dr. Abraham Rosenvasser”, in the framework of the Project PICT-Raíces 2015-2943, participated in the excavations at Tel Erani, Israel. Tel Erani, excavated since the 1950’s, is a key site to understand the relations between southern Palestine (Southern Levant) and Egypt during the Early Bronze IB (second half of the 4th millennium BC), since a significant presence of Egyptian findings has been found, including a sherd with a serekh of King Narmer. Furthermore, pottery of a characteristic local style, called Erani C has been found, identified also at the Nile Delta (Tell el-Farkha) and in proto-dynastic tombs, such as U-j in Abydos. Parts of these finds are related to at least two overlapping fortification walls that possibly surrounded the entire tel which occupied approximately 25 ha. This would be one of the earliest fortified settlements in southern Palestine. In the campaign of 2018, two areas were excavated: Area D3, where Egyptian material is abundant along with local elements, and Area P-Q, corresponding to one of the areas where the fortifications are located. The results of this last campaign indicate that these defensive walls would be prior to the Egyptian phase, i.e. during the Early Bronze IB1. During the late phase, Early Bronze IB2, the relations between both regions would have intensified, with the possibility that the Egyptians have played a more active role at Tel Erani