Search

Resumen La restricción hídrica es un aspecto limitante del cultivo del viñedo en muchas regiones, por lo que el riego constituye uno de los principales factores determinantes de su desarrollo en muchas zonas, que ha sido estudiado en diferentes ámbitos, pero la frecuencia de aplicación del riego, que afecta a su distribución en el suelo y puede influir en el comportamiento del viñedo, no ha sido suficientemente evaluada, por lo que su estudio resulta de gran interés en diversas condiciones de cultivo. El objetivo de este trabajo fue evaluar los efectos de cuatro frecuencias de riego deficitario del 30% ET0: 7 riegos por semana (T01), 2 riegos por semana (T03), 1 riego por semana (T07) y 1 riego cada dos semanas (T15), en la producción de uva y la composición de la baya. El experimento se desarrolló durante los años 2021 y 2022, en viñedos de Garnacha Tinta (en Badajoz: T03, T07, T15), Tempranillo (en Valladolid: T03, T07, T15), Syrah (en Albacete: T03 y T07) y Mencía (en Lugo: T01, T03, T07). Tanto la producción como la composición de la uva fueron determinadas en vendimia. Las diferentes frecuencias de riego han repercutido en el rendimiento y la calidad de la uva de forma variable según la variedad, la localización y el año. La producción de uva y la madera de poda se vieron ligeramente favorecidos por la frecuencia T07 con respecto a T03 en Garnacha y, sólo en la producción, en Tempranillo, mientras que ocurrió lo contrario en Syrah y Mencía. La concentración de azúcares se vio ligeramente favorecida por la frecuencia T07 en Syrah y, parcialmente, en Tempranillo. El pH del mosto no ofreció una respuesta apreciable derivada de la frecuencia de riego. La acidez total mostró valores ligeramente superiores de T07 en Garnacha y Tempranillo, pero algo más altos de T03 en Syrah y Mencía. El ácido tartárico apenas presentó una ligera tendencia favorable a T07 en Garnacha, mientras que el ácido málico mostró una tendencia ligeramente favorable a T07 en Garnacha y Mencía, pero beneficiosa para T03 en Syrah. El contenido de potasio y de polifenoles totales ofreció una clara variabilidad entre frecuencias de riego en general, aunque el primero mostró un ligero aumento del T03 con respecto al T07 en Mencía y el segundo un ligero aumento del T07 frente al T03 en Syrah. En definitiva, los efectos productivos y cualitativos derivados de la frecuencia de riego aplicada han tenido un alcance reducido, que, sin embargo, puede variar moderadamente en función de la variedad y del año.

Resumen El efecto sumidero de la uva frente a la superficie foliar, como fuente de fotosintatos, es un aspecto muy debatido en cuanto a la repercusión que puede tener en el desarrollo del viñedo y en la calidad de la uva. En ciertas zonas, la limitación del nivel productivo es argumentado como factor principal para alcanzar niveles de alta calidad en el vino, pero se conoce poco sobre los efectos que las distintas operaciones reguladoras pueden tener en variedades blancas de vid. Así, el trabajo persigue contrastar la respuesta de la cepa a la aplicación de dos alternativas de regulación de cosecha: aclareo de racimos y despampanado, aplicadas en la fase inmediatamente anterior al envero, en el cv. Verdejo, en España. Durante el periodo 2017-2019, la respuesta productiva y cualitativa del cv. Verdejo, sobre 110-R, fue estudiada tras aplicar, en fase de pre-envero, los tratamientos experimentales: AR, aclareo de racimos (respetando 1 racimo en cada pámpano); DP, despampanado (eliminando un pámpano en cada pulgar y respetando íntegramente el otro pámpano). El diseño experimental es en 4 bloques al azar, con parcela elemental de 12 cepas de control, en un viñedo conducido en espaldera, plantado en 2012, a 2,8 x 1,4 m, y podado en cordón Royat bilateral con 8 pulgares de 2 yemas por cepa. El desarrollo vegetativo se vio significativamente afectado por el despampanado con respecto al aclareo de racimos, reduciendo el peso de madera de poda casi un 40%, a pesar del aumento medio del 16% del peso del sarmiento. La producción de uva se vio reducida por el despampanado una media del 11%, como consecuencia, sobre todo, de la reducción del número de racimos por cepa, así como de la ligera disminución del peso del racimo. La concentración de azúcares, el pH, el ácido málico y el contenido de potasio se vieron ligeramente reducidos por el despampanado con respecto al aclareo, mientras que la acidez titulable y el ácido tartárico resultaron algo mayores. En definitiva, la eliminación de la mitad de los pámpanos, que conlleva la reducción de la relación fuente-sumidero con respecto al aclareo de racimos, ha limitado ligeramente el desarrollo de diversos componentes del rendimiento, así como la concentración de azúcares, de ácido málico y de potasio, a la vez que el pH, favoreciendo la acidez total en la uva. La elección de la técnica de regulación productiva dependerá de los aspectos cualitativos que se quieran potenciar y de la disponibilidad hídrica para el viñedo, pues el despampanado descarga más la expresión vegetativa, aliviando el posible estrés hídrico, que el aclareo directo de racimos.

La aplicación de riego al viñedo en zonas con balance hídrico muy deficiente se convierte en una necesidad a medida que avanza el verano y la escasa reserva hídrica del suelo se va agotando. El nivel de rehidratación del viñedo en dicha época puede afectar a su comportamiento fisiológico, productivo y cualitativo. A lo largo del periodo 2015-2018 se estudió la respuesta agronómica de 4 tratamientos derivados de la aplicación de riego de rehidratación, de 12 horas, a partir del envero, cuando se alcanza un nivel de potencial hídrico de tallo: secano (R0), cuando Ψx ≈ 1,7-1,8 MPa (R1), cuando Ψx ≈ 1,4-1,5 MPa (R2), cuando Ψx ≈ 1,0-1,2 MPa (R3). El ensayo se llevó a cabo con cv. Cabernet Sauvignon, plantado en 2004 y conducido en espaldera en cordón Royat bilateral, en Valladolid (España). El desarrollo vegetativo aumentó con el número de riegos de rehidratación, tal que el incremento medio del peso de madera de poda de R1, R2 y R3, respecto al secano (R0), fue de 13%, 28% y 50%, en estrecha correspondencia con el aumento del peso de sarmiento. La producción de uva también aumentó progresivamente con los riegos de rehidratación, con diferencias que discriminaron siempre el R3 del R0, e incrementos de R1, R2 y R3 de 18%, 33% y 44% con respecto a R0, habiéndose debido las diferencias mayormente al peso de racimo. La concentración de azúcares varió poco con el aumento de riegos de rehidratación, aunque el valor medio fue algo mayor en R1 y R0 que en R3 y R2, mientras que la acidez se vio favorecida, sobre todo en el tratamiento más regado, R3, ya que el pH re vio reducido a la vez que la acidez titulable, el ácido tartárico y el ácido málico aumentaron; siempre en detrimento más evidente del secano (R0). El potasio tendió al aumento en favor de R3, aunque no de forma constante a lo largo de los años.

La aplicación de riego en el viñedo se hace necesaria a lo largo del verano en muchas zonas de España, en la medida en que la reserva hídrica en el suelo se va agotando. El nivel de estrés hídrico soportado por la vid es un factor determinante de su respuesta fisiológica y, por tanto, productiva y cualitativa. A lo largo del periodo 2015-2017, en Valladolid, se estudió la aplicación de 4 regímenes hídricos, en cv. Cabernet Sauvignon, desde el envero a la madurez, basados en la rehidratación, de 12 horas de riego, según el potencial hídrico de tallo: no riego o secano (R0), cuando Ψx ≈ 1,7-1,8 MPa (R1), cuando Ψx ≈ 1,4-1,5 MPa (R2), cuando Ψx ≈ 1,0-1,2 MPa (R3). La hidratación inicial, efectuada en el envero excepto en R0, supuso una mejora inmediata del estado hídrico, que generó diferencias significativas de potencial frente al secano durante 3 ó 4 semanas. Los riegos de rehidratación posteriores, de R3 y R2, provocaron diferencias significativas entre ellos durante los intervalos entre riegos alternos, mientras que R1, que no se rehidrató, se aproximó gradualmente a R0. Las tasas de fotosíntesis, conductancia estomática y transpiración, medidas al cabo de un mes de la hidratación inicial, mostraron en general diferencias estadísticamente significativas entre los tratamientos de forma progresiva según el número de riegos recibidos, en orden decreciente: R3, R2, R1, R0. No obstante, las diferencias entre tratamientos a veces no resultaron estadísticamente significativas entre R3 y R2, ni entre R1 y R0, dependiendo del año de estudio.

La influencia del riego en zonas de clara escasez pluviométrica es conocida en general, pero la cantidad de agua aplicada en una zona de cultivo determinada puede afectar de modo particular a la producción y a la calidad de la uva. A lo largo del periodo 2012-2014 se estudió la respuesta agronómica a la aplicación de 3 tratamientos de régimen hídrico: secano (P0), riego del 25% ETo (P1) y riego del 50% ETo (P2), aplicado desde tamaño guisante hasta vendimia. El ensayo se llevó a cabo con cv. Cabernet Sauvignon sobre portainjerto 110R, plantado en 2004 y conducido en espaldera en cordón Royat bilateral, con marco de plantación de 2,2 m x 1,2 m, en Valladolid. El desarrollo vegetativo aumentó progresivamente con la dosis de riego, incrementándose el peso de madera de poda un 31% al pasar del tratamiento P0 al P1, y un 27% al pasar del P1 al P2, debido al peso del sarmiento. El rendimiento en uva también aumentó con las dosis de agua, un 25% al pasar del tratamiento P0 al P1 y un 13% al pasar del P1 al P2, debido mayormente al peso del racimo. La concentración de azúcares se vió ligeramente favorecida por el riego. El pH del mosto mostró cierta tendencia a la reducción con el riego, sobre todo con el de mayor dosis, a la par que la acidez titulable, el ácido tartárico y el ácido málico aumentaron con el riego, sobre todo con el de mayor dosis. El K y el IPT no mostraron diferencias notables ni constantes entre tratamientos, aunque en general el K aumentó algo con el riego y el IPT se redujo ligeramente con el riego de mayor dosis.

The knowledge of the behaviour of different grapevine rootstocks is basic to achieve a good adaptation of vine to its growing area. With the objective of knowing the agronomic and qualitative response of cv. Sauvignon Blanc to the use of several rootstocks, a trial was established in 2006 in the D.O. Rueda. The vines are vertical trellised, with bilateral Royat cordon pruning, and the vine spacing is 2.60 m × 1.25 m. The row orientation is NNW (N-25°). The experimental design consisted of randomized blocks with 4 replications and elemental plot of 60 vines. Over the period 2010–2014 it has been developed the study of the following 10 rootstocks (treatments): 110R, 101-14M, 420A, 3309C, 41B, 161-49C, 196-17C, Fercal, Gravesac and RGM. The rootstocks 420A and 41B showed a production higher than 50% with respect to 196-17C and 161-49C, and higher to 100% than RGM, due to the number of clusters per vine and, in greater amount, to the cluster weight. Fercal and Gravesac showed an increase of pruning wood weight of 24% with respect to 196-17C and 161-49C, and 90% with respect to RGM, mainly due to the shoot weight. The sugar concentration increased with 101-14M, 196-17C and Fercal, and became reduced with 161-49C, 41B and RGM. The pH of must was reduced with Fercal whereas the titratable acidity increased, which also showed increase with Gravesac and 161-49C. The tartaric acid hardly increased slightly with Fercal and 161-49C, whereas the malic acid increased with Gravesac and Fercal, and was reduced with 41B, 3309C, RGM and 101-14M. The potassium concentration increased with 196-17C, Gravesac and Fercal, and was reduced with 41B, 161-49C, 420A and 3309C. The effects observed show alternatives for rootstock election according to the growing conditions and objectives of the vineyard.

The control of grape yield can be a critical aspect for grape quality of white varieties, moreover in the conditions of high water limitation that occurs in most Spanish growing regions. In the case of Verdejo variety, it is necessary to know its response to the reduction of the number of clusters per vine in order to regulate the production and improve the grape quality. During the period 2012–2014, the application of cluster thinning has been studied, by means of a treatment (A) of removing 27% of clusters, in comparison with a reference treatment (T) without thinning. The trial has been developed with vines of cv. Verdejo on 110R rootstock, planted in 2006 and trellised on bilateral Royat cordon. Vine distances were 2.60 m × 1.25 m (3077 plants/ ha). Vineyard growing was developed under rainfed conditions in Medina del Campo (Valladolid, Spain), within the D.O. Rueda. The water and physiological response of vines subjected to thinning has been similar to that of the reference vines. The vegetative growth has not shown significant differences between treatments. However, as expected, the cluster thinning resulted in a substantial reduction of grape yield as a direct consequence of the removal of clusters, although the cluster weight was slightly favored by cluster thinning. The quality of grapes has been modified by cluster thinning, in such a way that the sugar concentration has benefited, while the titratable acidity and tartaric acid have been reduced. However, the malic acid and the pH have hardly been affected by cluster thinning, whereas the potassium concentration has clearly increased as a result of cluster thinning.

This paper studies the vegetative, productive and qualitative behavior of the Tempranillo variety, vertically trellised trained, according to a placement of vines with three different distances (1.04, 1.40 and 1.80 meters) in the row and a common distance between rows (2.40 meters), in order to determine the most suitable vine distance for the growing conditions. The experimental test has been developed for the period 2007–2011 in the Appellation of Origin Cigales, in Valladolid (Spain). The reduction of vine distance has favored the vegetative growth, through the shoot weight, while the grape yield has been slightly higher in the intermediate vine distance, 1.40 m, through the cluster weight. Ravaz index showed some increase with the increasing of vine distance, derived from the decrease of pruning wood. In qualitative terms, the variation of the vine distance did not significantly modify the basic composition of the grape. Therefore, the choice of the vine distance involves the convenience to assess both potential effects in terms of production and economic objectives of the new plantations, in accordance with the characteristics of each growing situation.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a chronic liver condition that often progresses to more advanced stages, such as metabolic dysfunction-associated steatohepatitis (MASH). MASH is characterized by inflammation and hepatocellular ballooning, in addition to hepatic steatosis. Despite the relatively high incidence of MASH in the population and its potential detrimental effects on human health, this liver disease is still not fully understood from a pathophysiological perspective. Deregulation of polyamine levels has been detected in various pathological conditions, including neurodegenerative diseases, inflammation, and cancer. However, the role of the polyamine pathway in chronic liver disorders such as MASLD has not been explored. In this study, we measured the expression of liver ornithine decarboxylase (ODC1), the rate-limiting enzyme responsible for the production of putrescine, and the hepatic levels of putrescine, in a preclinical model of MASH as well as in liver biopsies of patients with obesity undergoing bariatric surgery. Our findings reveal that expression of ODC1 and the levels of putrescine, but not spermidine nor spermine, are elevated in hepatic tissue of both diet-induced MASH mice and patients with biopsy-proven MASH compared with control mice and patients without MASH, respectively. Furthermore, we found that the levels of putrescine were positively associated with higher aspartate aminotransferase concentrations in serum and an increased SAF score (steatosis, activity, fibrosis). Additionally, in in vitro assays using human HepG2 cells, we demonstrate that elevated levels of putrescine exacerbate the cellular response to palmitic acid, leading to decreased cell viability and increased release of CK-18. Our results support an association between the expression of ODC1 and the progression of MASLD, which could have translational relevance in understanding the onset of this disease. © 2024 The Pathological Society of Great Britain and Ireland., (© 2024 The Pathological Society of Great Britain and Ireland.)

Urolithins are gut microbiota metabolites of ellagic acid. Here, we have identified and chemically characterized a novel urolithin produced from urolithin D (3,4,8,9-tetrahydroxy urolithin) by in vitro incubation with different human gut Enterocloster species under anaerobic conditions. Urolithin G (3,4,8-trihydroxy urolithin) was identified by 1 H NMR, 13 C NMR, UV, HRMS, and 2D NMR. For the identification, NMR spectra of other known urolithins were also recorded and compared. Urolithin G was present in the feces of 12% of volunteers in an overweight-obese group after consuming an ellagitannin-rich pomegranate extract. The production of urolithin G required a bacterial 9-dehydroxylase activity and was not specific to the known human urolithin metabotypes A and B. The ability to produce urolithin G could be considered an additional metabolic feature for volunteer stratification and bioactivity studies. This is the first urolithin with a catechol group in ring A while having only one hydroxyl in ring B, a unique feature not found in human and animal samples so far.

Nitric oxide (NO) plays a pivotal role in integrating dopamine transmission in the basal ganglia and has been implicated in the pathogenesis of Parkinson disease (PD). The objective of this study was to ascertain whether the NO synthase inhibitor, 7-nitroindazole (7-NI), is able to reduce L-DOPA-induced dyskinesias (LIDs) in a non-human primate model of PD chronically intoxicated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Six Parkinsonian macaques were treated daily with L-DOPA for 3-4 months until they developed LIDs. Three animals were then co-treated with a single dose of 7-NI administered 45 min before each L-DOPA treatment. Dyskinetic MPTP-treated monkeys showed a significant decrease in LIDs compared with their scores without 7-NI treatment ( p < 0.05). The anti-Parkinsonian effect of L-DOPA was similar in all three monkeys with and without 7-NI co-treatment. This improvement was significant with respect to the intensity and duration of LIDs while the beneficial effect of L-DOPA treatment was maintained and could represent a promising therapy to improve the quality of life of PD patients.

The impact of age-associated disorders is increasing as the life expectancy of the population increments. Cardiovascular diseases and neurodegenerative disorders, such as Parkinson's disease, have the highest social and economic burden and increasing evidence show interrelations between them. Particularly, dysfunction of the cardiovascular nervous system is part of the dysautonomic symptoms of Parkinson's disease, although more studies are needed to elucidate the role of cardiac function on it. We analyzed the dopaminergic system in the nigrostriatal pathway of Parkinsonian and dyskinetic monkeys and the expression of some key proteins in the metabolism and synthesis of catecholamines in the heart: total and phosphorylated (phospho) tyrosine hydroxylase (TH), and membrane (MB) and soluble (S) isoforms of catechol-O-methyl transferase (COMT). The dopaminergic system was significantly depleted in all MPTP-intoxicated monkeys. MPTP- and MPTP + L-DOPA-treated animals also showed a decrease in total TH expression in both right (RV) and left ventricle (LV). We found a significant increase of phospho-TH in both groups (MPTP and MPTP + L-DOPA) in the LV, while this increase was only observed in MPTP-treated monkeys in the RV. MB-COMT analysis showed a very significant increase of this isoform in the LV of MPTP- and MPTP + L-DOPA-treated animals, with no significant differences in S-COMT levels. These data suggest that MB-COMT is the main isoform implicated in the cardiac noradrenergic changes observed after MPTP treatment, suggesting an increase in noradrenaline (NA) metabolism. Moreover, the increase of TH activity indicates that cardiac noradrenergic neurons still respond despite MPTP treatment., (© 2021. The Author(s).)

Community-acquired pneumonia (CAP) continues to be an important cause of morbidity and mortality in adults. The aim of this study is to update the practical prevention guide for CAP through vaccination in Spain developed in 2016 and updated in 2018, based on available vaccines and evidence through bibliographic review and expert opinion. The arrival of COVID-19 as a new cause of CAP and the recent availability of safe and effective vaccines constitutes the most significant change. Vaccines against pneumococcus, influenza, pertussis and COVID-19 can help to reduce the burden of disease from CAP and its associated complications. The available evidence supports the priority indications established in this guide, and it would be advisable to try to achieve a widespread dissemination and implementation of these recommendations in routine clinical practice., (Copyright © 2021 Sociedad Española de Médicos de Atención Primaria (SEMERGEN). Publicado por Elsevier España, S.L.U. All rights reserved.)

Temperate fruit trees belonging to Prunus species have the ability to suspend (induce dormancy) and resume growth periodically in response to environmental and seasonal conditions. Endodormancy release requires the long-term accumulation of chill. Upon accumulation of cultivar-specific chill requirements, plants enter the state of ecodormancy, which means the ability to grow has been restored, depending on the fulfilment of heat requirements. As many different metabolic pathways are implicated in endodormancy release, we have performed a metabolomic analysis, using the ultra-high-performance liquid chromatography-quadrupole time-of-flying (UPLC-QToF) technique. We assayed flower buds in different stages of endodormancy in four almond cultivars with different flowering times: the extra-early Desmayo Largueta, the late Antoñeta, the extra-late Penta, and the ultra-late Tardona. An orthogonal projection to latent-structure discriminant-analysis model was created to observe differences between endodormant and ecodormant flower buds. The metabolites showing the most significant variation were searched against the Metlin, HMDB, and KEGG libraries, which allowed us to identify 87 metabolites. These metabolites were subsequently assigned to specific pathways, such as abscisic acid biosynthesis, phenylpropanoid biosynthesis, and D-sorbitol metabolism, among others. The two metabolites that exhibited the most significant variations in all the cultivars studied with fold changes of up to 6.49 were ascorbic acid and prunasin. For the first time, these two metabolites have been proposed as potential biomarkers for endodormancy release in almond. Given the high synteny present between the Rosaceae species, these results could be extrapolated to other important crops like peach, plum, cherry, or apricot, among others.

Background: Autonomic dysfunction is a well-known dominant symptom in the advanced stages of Parkinson's disease. However, the role of cardiac sympathetic nerves still needs to be elucidated., Objectives: To evaluate cardiac sympathetic response in Parkinsonian and dyskinetic monkeys., Methods: Adult male monkeys were divided into 1 of the following 3 groups: controls, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated monkeys, and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine+levodopa-treated animals. Noradrenaline, its metabolite normetanephrine, and phospho-Heat shock proten 27 (p-Hsp27) at serine 82 levels were analyzed in the left and right ventricles of the heart. Tyrosine hydroxylase immunohistochemistry was performed in the ventral mesencephalon., Results: The results were the following: (1) 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine intoxication significantly increased normetanephrine levels and decreased noradrenaline turnover in the right ventricle without changes in the left ventricle; however, (2) levodopa treatment decreased noradrenaline levels and enhanced the normetanephrine/noradrenaline ratio in parallel with a very significant increase of Hsp27 activity in both ventricles., Conclusions: Levodopa treatment could induce protective cardiac effects through the increased Hsp27 activity. © 2019 International Parkinson and Movement Disorder Society., (© 2019 International Parkinson and Movement Disorder Society.)

Streptococcus pneumoniae shows more than 90 capsular serotypes that can be distinguished by their reactivity against antisera. The main objective of this work was the development of a molecular method for serotyping without the use of antisera. A computer program containing an algorithm was used to search in a database for potentially useful enzymes for Restriction Fragment Length Polymorphism-RFLP typing, in order to maximize the discrimination between different serotypes. DNA sequences of 90 serotypes for the region between dex B and ali A genes were compiled, and a computer screening of restriction enzymes was performed. The wzg - wzh - wzd - wze region and Sse 9I restriction predicted unique PCR-RFLP patterns for 39 serotypes and eight serogroups. A second restriction enzyme resolved fragment specific patterns for 25 serotypes. The method was tested with 98 serotype-unknown clinical isolates. PCR-RFLP analysis deduced correct serotypes that were confirmed by Quellung reaction for 78.5% of the isolates., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Paroxysmal Nocturnal Hemoglobinuria (PNH) is a clonal disease of blood cells caused by the lack of glycosyl phosphatidyl inositol anchored proteins bound to the cell membrane. In consequence, erythrocytes lead to intravascular hemolysis upon complement activation, which promotes high risk of thrombosis, intravascular hemolytic anemia, and bone marrow failure in patients. The mechanisms of thrombosis in PNH are still poorly understood. Treatment with eculizumab reduces intravascular hemolysis and thrombotic risk, but not in all cases. Exosomes are extracellular vesicles released by cells and whose secretion is closely related to the inflammatory status. They participate in cell communication by activating signaling pathways and transferring genetic material and proteins to host cells. In consequence, exosomes may serve as surrogate biomarkers for the prognosis and/or diagnosis of a disease. Isolation of exosomes was carried out from healthy controls and from three groups of PNH patients, i.e. i) with no eculizumab treatment; ii) under treatment with eculizumab that have not suffered thrombosis; and iii) under treatment with eculizumab but that have suffered thrombosis. The miRNAome and proteome was analyzed using plasma focus miRNAs PCR panel and LC-MS analysis respectively. We found differential expression of miRNAs miR-148b-3p, miR-423-3p, miR29b-3p, miR15b-5p, let-7e-5p, miR126-3p, miR-125b-5p and miR-376c-3p as well as hemoglobin, haptoglobin, protein S and C4-binding protein in healthy controls vs PNH patients. Our results warrant further research and provide new information on the content of exosomes that could play a role in the hypercoagulable state in this disease.

Aim: To test the synergistic effect of Cpl-711 endolysin and antibiotics for antipneumococcal activity., Materials & Methods: A combination of Cpl-711 and different antibiotics (amoxicillin, cefotaxime, levofloxacin and vancomycin) was tested in a checkerboard assay against several multidrug-resistant Streptococcus pneumoniae strains. Mouse and zebrafish models of pneumococcal sepsis were used to confirm the in vitro data., Results: The activity of Cpl-711 combined with amoxicillin or cefotaxime was synergistic in the bactericidal effect against a serotype 23F multiresistant clinical isolate of S. pneumoniae. Synergy between Cpl-711 and cefotaxime was validated using both mouse and zebrafish models., Conclusion: Combination of Cpl-711 and cefotaxime may help in the treatment of diseases caused by multiresistant pneumococcal strains.

Parkinson's disease (PD) is an age-related neurodegenerative disease associated with the misfolding and aggregation of alpha-synuclein (aSyn). The molecular underpinnings of PD are still obscure, but nutrition may play an important role in the prevention, onset, and disease progression. Dietary (poly)phenols revert and prevent age-related cognitive decline and neurodegeneration in model systems. However, only limited attempts were made to evaluate the impact of digestion on the bioactivities of (poly)phenols and determine their mechanisms of action. This constitutes a challenge for the development of (poly)phenol-based nutritional therapies. Here, we subjected (poly)phenols from Arbutus unedo to in vitro digestion and tested the products in cell models of PD based on the cytotoxicity of aSyn. The (poly)phenol-digested metabolites from A. unedo leaves (LPDMs) effectively counteracted aSyn and H 2 O 2 toxicity in yeast and human cells, improving viability by reducing aSyn aggregation and inducing its clearance. In addition, LPDMs modulated pathways associated with aSyn toxicity, such as oxidative stress, endoplasmic reticulum (ER) stress, mitochondrial impairment, and SIR2 expression. Overall, LPDMs reduced aSyn toxicity, enhanced the efficiency of ER-associated protein degradation by the proteasome and autophagy, and reduced oxidative stress. In total, our study opens novel avenues for the exploitation of (poly)phenols in nutrition and health.

Airway infection by nontypeable Haemophilus influenzae (NTHi) associates to chronic obstructive pulmonary disease (COPD) exacerbation and asthma neutrophilic airway inflammation. Lipids are key inflammatory mediators in these disease conditions and consequently, NTHi may encounter free fatty acids during airway persistence. However, molecular information on the interplay NTHi-free fatty acids is limited, and we lack evidence on the importance of such interaction to infection. Maintenance of the outer membrane lipid asymmetry may play an essential role in NTHi barrier function and interaction with hydrophobic molecules. VacJ/MlaA-MlaBCDEF prevents phospholipid accumulation at the bacterial surface, being the only system involved in maintaining membrane asymmetry identified in NTHi. We assessed the relationship among the NTHi VacJ/MlaA outer membrane lipoprotein, bacterial and exogenous fatty acids, and respiratory infection. The vacJ/mlaA gene inactivation increased NTHi fatty acid and phospholipid global content and fatty acyl specific species, which in turn increased bacterial susceptibility to hydrophobic antimicrobials, decreased NTHi epithelial infection, and increased clearance during pulmonary infection in mice with both normal lung function and emphysema, maybe related to their shared lung fatty acid profiles. Altogether, we provide evidence for VacJ/MlaA as a key bacterial factor modulating NTHi survival at the human airway upon exposure to hydrophobic molecules.

Antibacterial treatment with cotrimoxazol (TxS), a combination of trimethoprim and sulfamethoxazole, generates resistance by, among others, acquisition of thymidine auxotrophy associated with mutations in the thymidylate synthase gene thyA , which can modify the biology of infection. The opportunistic pathogen non-typeable Haemophilus influenzae (NTHi) is frequently encountered in the lower airways of chronic obstructive pulmonary disease (COPD) patients, and associated with acute exacerbation of COPD symptoms. Increasing resistance of NTHi to TxS limits its suitability as initial antibacterial against COPD exacerbation, although its relationship with thymidine auxotrophy is unknown. In this study, the analysis of 2,542 NTHi isolates recovered at Bellvitge University Hospital (Spain) in the period 2010-2014 revealed 119 strains forming slow-growing colonies on the thymidine low concentration medium Mueller Hinton Fastidious, including one strain isolated from a COPD patient undergoing TxS therapy that was a reversible thymidine auxotroph. To assess the impact of thymidine auxotrophy in the NTHi-host interplay during respiratory infection, thyA mutants were generated in both the clinical isolate NTHi375 and the reference strain RdKW20. Inactivation of the thyA gene increased TxS resistance, but also promoted morphological changes consistent with elongation and impaired bacterial division, which altered H. influenzae self-aggregation, phosphorylcholine level, C3b deposition, and airway epithelial infection patterns. Availability of external thymidine contributed to overcome such auxotrophy and TxS effect, potentially facilitated by the nucleoside transporter nupC . Although, thyA inactivation resulted in bacterial attenuation in a lung infection mouse model, it also rendered a lower clearance upon a TxS challenge in vivo . Thus, our results show that thymidine auxotrophy modulates both the NTHi host airway interplay and antibiotic resistance, which should be considered at the clinical setting for the consequences of TxS administration.

Nitric oxide (NO) is a pleiotropic janus-faced molecule synthesized by nitric oxide synthases (NOS) which plays a critical role in a number of physiological and pathological processes in humans. The physiological roles of NO depend on its local concentrations, as well as its availability and the nature of downstream target molecules. Its double-edged sword action has been linked to neurodegenerative disorders. Excessive NO production, as the evoked by inflammatory signals, has been identified as one of the major causative reasons for the pathogenesis of several neurodegenerative diseases. Moreover, excessive NO synthesis under neuroinflammation leads to the formation of reactive nitrogen species and neuronal cell death. There is an intimate relation between microglial activation, NO and neuroinflammation in the human brain. The role of NO in neuroinflammation has been defined in animal models where this neurotransmitter can modulate the inflammatory process acting on key regulatory pathways, such as those associated with excitotoxicity processes induced by glutamate accumulation and microglial activation. Activated glia express inducible NOS and produce NO that triggers calcium mobilization from the endoplasmic reticulum, activating the release of vesicular glutamate from astroglial cells resulting in neuronal death. This change in microglia potentially contributes to the increased age-associated susceptibility and neurodegeneration. In the current review, information is provided about the role of NO, glial activation and age-related processes in the central nervous system (CNS) that may be helpful in the isolation of new therapeutic targets for aging and neurodegenerative diseases.

Deep brain stimulation (DBS) has become the standard surgical procedure for advanced Parkinson's disease (PD). Recently, the pedunculopontine tegmental nucleus (PPN) has emerged as a potential target for DBS in patients whose quality of life is compromised by freezing of gait and falls. To date, only a few groups have published their long-term clinical experience with PPN stimulation. Bearing in mind that the Barrington (Bar) nucleus and some adjacent nuclei (also known as the micturition centre) are close to the PPN and may be affected by DBS, the aim of the present study was to review the anatomical location of this structure in human and other species. To this end, the Bar nucleus area was analysed in mouse, monkey and human tissues, paying particular attention to the anatomical position in humans, where it has been largely overlooked. Results confirm that anatomical location renders the Bar nucleus susceptible to influence by the PPN DBS lead or to diffusion of electrical current. This may have an undesirable impact on the quality of life of patients., (Copyright © 2013 IBRO. All rights reserved.)

Patients with Parkinson's disease show persistent microglial activation in the areas of the brain where the degeneration of dopaminergic neurons takes place. The reason for maintaining this activated state is still unknown, but it is thought that this persistent microglial activation may contribute to the degeneration of dopaminergic neurons. In this study, we report the microanatomical details of microglia and the relationship between microglia and neurons in the substantia nigra pars compacta of Parkinsonian monkeys years after insult with MPTP. We observed that microglial cells appear polarized toward dopaminergic neurons in MPTP-treated macaques compared to untreated animals and present clear phagocytic characteristics, such as engulfing gliaptic contacts, an increase in Golgi apparatus protein machinery and ball-and-chain phagocytic buds. These results demonstrate that activated microglia maintain phagocytic characteristics years after neurotoxin insult, and phagocytosis may be a key contributor to the neurodegenerative process., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Neuronal nitric oxide synthase (nNOS) is involved in the regulation of diverse intracellular messenger systems in the brain. Nitric Oxide (NO) contributes to inducing signaling cascades that involve a complex pattern of phosphorylation of DARPP-32 (in Thr-34), which controls the phosphoproteins involved in neuronal activation. However, the role of NO in the pathophysiology of Parkinson's disease (PD) and its effect in striatal neurons have been scarcely explored. In the present work, we investigate the effects of a nitric oxide synthase (NOS) inhibitor, 7-nitroindazole (7-NI) in the nigrostriatal pathway of striatal 6-hydroxydopamine (6-OHDA) lesioned rats. Our quantitative histological findings show that treatment with 7-NI significantly reduced 6-OHDA-induced dopaminergic damage in the dorsolateral striatum and Substantia Nigra pars compacta (SNpc). Moreover, 6-OHDA lesioned rats show a significant increase of nNOS(+) and Phospho-Thr34-DARPP-32(+) cells, accompanied by a consequent decrease of total DARPP-32(+) cells, which suggests an imbalance of NO activity in the DA-depleted striatum, which is also reflected in behavioral studies. Importantly, these effects are reverted in the group treated with 7-NI. These results show a clear link between the state of phosphorylation of DARPP-32 and parkinsonism, which is regulated by nNOS. This new evidence suggests a prominent role for nitric oxide in the neurotransmitter balance within the basal ganglia in the pathophysiology of experimental parkinsonism., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

In the present work we analyze the cerebellum of chronic parkinsonian monkeys in order to clarify whether chronic mesencephalic depletion is associated with long term activation of the cerebellar neurons in chronic Parkinsonism. In our study, we observed a persistent activation of Purkinje cells in the cerebellum of chronic parkinsonian macaques, characterized by the expression of c-Fos, which correlated with dopaminergic degeneration. These results are compatible with the results observed in fMRI in Parkinson's disease patients, and may contribute to the understanding of additional alterations in the brain circuitry in Parkinsonism.

The role of microglial motility in the context of adult neurodegeneration is poorly understood. In the present work, we investigated the microanatomical details of microglia-neuron interactions in an experimental mouse model of Parkinson's disease following the intraperitoneal injection of MPTP. The specific intoxication of dopaminergic neurons induces the cellular polarization of microglia, leading to the formation of body-to-body neuron-glia contacts, called gliapses, which precede neuron elimination. Inhibiting ROCK/Cdc42-mediated microglial motility in vivo blocks the activating features of microglia, such as increased cell size and number of filopodia and diminishes their phagocyting/secreting domains, as the reduction of the Golgi apparatus and the number of microglia-neuron contacts has shown. High-resolution confocal images and three-dimensional rendering demonstrate that microglia engulf entire neurons at one-to-one ratio, and the microglial cell body participates in the formation of the phagocytic cup, engulfing and eliminating neurons in areas of dopaminergic degeneration in adult mammals.