Serological Determination of West Nile Virus in Domestic Birds from Rapa Nui, Chile.

Background: Flaviviruses are agents with high zoonotic potential of importance to human health. They are transmitted by mosquitoes of the Culicidae family, and birds act as host-amplifiers. Birds, mammals, and humans are susceptible hosts to infection. Methods: In this study, West Nile virus (WNV), flavivirus, infection was studied in 37 serum samples from 22 hens on Easter Island, Chile. Results: WNV was detected by ELISA (ID Screen® West Nile Competition Multi-Species). We report absence of antibodies to WNV, and to related viruses of the Japanese Encephalitis Virus serocomplex, and, therefore, absence of infection across the sample. Conclusion: This is the first evaluation of its type carried out in Chile, and represents a positive result for public health at Easter Island.

A g-type lysozyme from the scallop Argopecten purpuratus participates in the immune response and in the stability of the hemolymph microbiota.

Lysozymes are antimicrobial acid hydrolases widely distributed in nature. They are located inside the cells in lysosomes, or they are secreted to the extracellular space, where they can lyse the cell wall of certain species of bacteria via hydrolysis of the peptidoglycan. Thus, lysozymes are bacteriolytic enzymes and play a major biological role in biodefense, as these enzymes can act as antibacterial and immune-modulating agents. In this study, we characterized a g-type lysozyme from the scallop Argopecten purpuratus named ApGlys. The cDNA sequence comprises an open reading frame (ORF) of 600 nucleotides, codifying for a putative protein of 200 amino acids with a signal peptide of 18 amino acids. The deduced mature protein sequence displays a molecular weight of 20.07 kDa and an isoelectric point (pI) of 6.49. ApGlys deduced protein sequence exhibits conserved residues associated with catalytic activity and substrate fixation in other g-type lysozymes. The phylogenetic analysis revealed a high degree of identity of ApGlys with other mollusk g-type lysozymes, which form a restricted and separated clade from the vertebrate lysozymes. ApGlys transcripts were constitutively and highly expressed in the digestive gland, and it was induced in hemocytes and gills of scallops after an immune challenge. Furthermore, the ApGlys protein was located inside hemocytes of immunostimulated scallops, determined by immunofluorescence analysis. Finally, the transcript silencing of ApGlys by RNA interference led to an increase of total culturable bacteria from the scallop hemolymph. Furthermore, we detected a higher diversity of the bacterial community in ApGlys-silenced scallops and an imbalance of certain bacterial groups present in the hemolymph by 16S rDNA deep amplicon sequencing. Overall, our results showed that ApGlys is a new member of scallop lysozymes that is implicated in the immune response and in the microbial homeostasis of A. purpuratus hemolymph.

Evaluation of extracellular vesicles and gDNA from culture medium as a possible indicator of developmental competence in human embryos.

Human embryos generated in vitro have a high incidence of chromosomal abnormalities that negatively affect pregnancy rate. Embryos generated in vitro secrete extracellular vesicles (EVs) into the culture medium that could be used potentially as indicators of embryo competence. This research aimed to evaluate the concentration and size of EVs and their gDNA content as an indicator of developmental competence in human embryos. Human embryos generated by intracytoplasmic sperm injection (ICSI) were classified morphologically as of either TOP, FAIR or POOR quality. Culture medium and developmentally arrested embryos (which were not able to be used for embryo transfer) were collected. Microvesicles, exosomes (MV/Exo) and apoptotic bodies (ABs) were isolated from culture medium. Nanoparticle tracking analysis (NTA) and array comparative genomic hybridization (aCGH) analysis were performed to evaluate EVs and their gDNA content. From NTA, the diameter (mean) of MVs/Exo from TOP quality embryos was higher (112.17 nm) compared with that of FAIR (108.02) and POOR quality embryos (102.78 nm) (P < 0.05). aCGH analysis indicated that MVs/Exo and ABs carried gDNA with the presence of 23 chromosome pairs. However, when arrested embryos were compared with their respective MVs/Exo and ABs, the latter had an increased rate of chromosomal abnormalities (24.9%) compared with embryos (8.7%) (P < 0.05). In conclusion, the size of EVs from culture medium might be an alternative for evaluating competence of human embryos, however more studies are needed to validate the use of gDNA from EVs as an indicator of embryo competence.

The Influence of pCO2-Driven Ocean Acidification on Open Ocean Bacterial Communities during A Short-Term Microcosm Experiment in the Eastern Tropical South Pacific (ETSP) off Northern Chile.

Due to the increasing anthropogenic CO2 emissions, Ocean Acidification (OA) is progressing rapidly around the world. Despite the major role that microorganisms play on the marine biogeochemical cycling and ecosystem functioning, the response of bacterial communities upon OA scenarios is still not well understood. Here, we have conducted a detailed characterization of the composition and relative abundance of bacterial communities in the water column of an open-ocean station in the Eastern Tropical South Pacific (ETSP) off northern Chile and their interactions with environmental factors. In addition, through a short-term microcosm experiment, we have assessed the effect of low pH/high pCO2 conditions over the abundance and genetic diversity of bacterial communities. Our results evidence a clear partitioning of community composition that could be attributed mostly to dissolved oxygen. However, our experimental approach demonstrated that low pH/high pCO2 conditions might modify the structure of the bacterial community, evidencing that small changes in pH may impact significantly the abundance and diversity of key microorganisms. This study constitutes a first step aiming to provide insight about the influence of changing carbonate chemistry conditions on natural bacterial communities and to shed light on the potential impact of OA in biogeochemical cycles on the ETSP region.

Tennis elbow: associated psychological factors.

BACKGROUND: The etiology of tennis elbow is multifactorial. Overuse of the wrist extensors along with anatomic factors, such as flexibility problems, aging, and poor blood circulation, may play a role. This study investigated whether patients with tennis elbow have a different psychological profile compared with healthy controls. METHODS: Patients with clinical signs of tennis elbow, consulting at the Ghent University Hospital between September 2015 and January 2017, were offered a paper-and-pencil questionnaire about Big Five personality traits, perfectionism, anxiety, depression, work satisfaction, and working conditions. Healthy controls in the same risk group were offered the same questionnaires. RESULTS: We recruited 69 patients (35 men, 34 women) and 100 controls (44 men, 56 women). Tennis elbow patients scored significantly lower on the personality traits extraversion and agreeableness. Men, in particular, scored significantly higher on perfectionism and were more likely to develop an anxiety disorder or a depression. Concerning work, patients indicated a significantly higher workload (especially men) and a significantly lower autonomy (especially women). Female patients also indicated less contact with colleagues. However, work satisfaction was relatively high in both groups. CONCLUSION: The results suggest that there is a relationship between complaints related to tennis elbow and psychological characteristics.

Adaptación y validación de un cuestionario grupal sobre participación comunitaria en salud en la comuna de El Bosque, Santiago de Chile / Adaptation and validation of a group questionnaire on community health participation in el bosque municipality, Santiago, Chile

La participación comunitaria ha sido uno de los aspectos fundamentales en las políticas sanitarias en Chile desde 1990. Sin embargo, aun reconociéndose la importancia de la participación comunitaria en el trabajo de los equipos de salud, esta estrategia ha presentado dificultades en su implementación, derivadas tanto de aspectos prácticos como de problemas teóricos. La ausencia de definiciones conceptuales claras y compartidas acerca de la participación resulta en tensiones ideológicas y políticas que se expresan a su vez en una interpretación, aplicación y evaluación distinta de las actividades de participación. Así, se hace necesario desarrollar herramientas de evaluación de la participación que, utilizándose transversalmente, fortalezcan el trabajo de los equipos de salud, al mismo tiempo que promuevan el desarrollo teórico del área. Un instrumento de evaluación de la participación comunitaria, diseñado especialmente para ser utilizado en atención primaria de salud, lo constituye el Spidergram, o mapa en tela de araña, que incorpora la evaluación de cinco dimensiones del proceso participativo. El presente estudio tiene por objetivo adaptar y validar este instrumento en un contexto urbano en Chile. Se evaluó la validez del cuestionario grupal traducido al español basado en la revisión de la literatura, la aplicación del instrumento piloto y las discusiones sostenidas por parte del equipo de investigación. Este instrumento alcanzó un adecuado nivel de consistencia interna y de concordancia interobservador en la mayoría de las variables al ser aplicado por moderadores externos, permitiéndose su recomendación como herramienta de trabajo para los equipos de atención primaria de salud.

Community participation has been one of the fundamental aspects of Chilean health care policies since 1990. Though the importance of health care teams' community participation should be recognized, this strategy has nevertheless faced a number of obstacles during its implementation, due to practical and theoretical problems. The absence of a clear and common conceptual definition of community participation results in ideological and political tensions that are, in turn, expressed in different interpretations, applications, and assessments of participatory activities. Thus, it is necessary to develop methods for the assessment of community participation that can be used across the board, and which may allow health care workers to strengthen their work with local communities, while promoting theoretical advances in the area. One instrument for measuring community participation, especially designed to be used in primary health care settings, is the Spidergram, which includes the evaluation of 5 variables related to the participatory process. This study aims to adapt and validate this assessment method in an urban context in Chile. The validity of the instrument was assessed through a literature review, a pilot application of a version translated into Spanish, and multiple discussions within the multidisciplinary research team. This instrument achieved adequate internal consistency and inter-observer concordance in most variables when applied by external observers, supporting its recommendation as a valuable tool for primary health care workers.

Prevalencia de pacientes con tratamiento sustitutivo renal en El Salvador en 2014 / Prevalence of patients receiving renal replacement therapy in El Salvador in 2014

El Salvador presenta la mortalidad por fallo renal más alta de América. Cinco proveedores de servicios de salud ofrecen tratamiento sustitutivo renal (TSR) en el país. La prevalencia nacional de TSR nunca ha sido reportada. Objetivos: Conocer la prevalencia de TSR en El Salvador y algunas características básicas. Métodos: La asociación de nefrología coordinó una encuesta nacional transversal durante el tercer trimestre de 2014; 31 centros renales participaron, que cubrían el 99,5% de los pacientes. Resultados: Prevalencia nacional de TSR: 595 por millón de población (pmp), N=3.807, edad promedio 50,4 años, 67,5% hombres. Por modalidad: diálisis peritoneal (DP) 289 pmp; hemodiálisis (HD) 233 pmp; con trasplante renal funcionante 74 pmp (donante vivo solamente). La seguridad social cubre el 25% de la población, pero atiende al 49,7% de los pacientes en TSR. Generalmente observamos prevalencias más altas en municipios con centros renales, costeros o de zonas bajas. Un 95% de los pacientes en HD reciben menos de 3 sesiones semanales. Un 59% de los pacientes en DP no están en continua ambulatoria o programas automatizados y un 25% todavía utilizan catéter rígido. Etiología de la enfermedad renal crónica: no reportada/desconocida 50%, hipertensión 21,1%, diabetes 18,9%, glomerulonefritis 6,7%, obstructivas 1,2%, tubulointersticiales 0,9%, poliquísticas 0,4%, otras 0,7%. Discusión: A pesar del incremento de servicios de TSR, la prevalencia es inferior al promedio latinoamericano (660 pmp). Un 75% de los pacientes en HD y DP están infradializados. Todavía se utilizan técnicas anticuadas. La presencia de la nefropatía mesoamericana influye las características demográficas (muchos jóvenes, 2/3 hombres, alta prevalencia en zonas bajas y costeras) (AU)

El Salvador has the highest renal failure mortality rate in the Americas. Five healthcare providers offer renal replacement therapy (RRT) in the country. The national RRT prevalence has never been reported. Aims: To determine the RRT prevalence in El Salvador and some basic characteristics. Methods: The association of nephrology coordinated a nationwide cross-sectional survey during the third quarter of 2014. 31 renal centres participated in the survey, covering 99.5% of patients. Results: National RRT prevalence: 595 per million population (pmp), N=3807, average age 50.4 years, 67.5% male. By modality: peritoneal dialysis (PD) 289 pmp, haemodialysis (HD) 233 pmp, with functioning kidney transplantation 74 pmp (living donor only). Social security covers 25% of the population but treats 49.7% of RRT patients. Generally, higher prevalence was observed in municipalities with renal centres or located on the coast or lowlands. Ninety-five percent of HD patients receive fewer than 3 weekly sessions. Of PD patients, 59% do not belong to a continuous outpatient or automated programme, and 25% still use rigid catheter. Aetiology of chronic kidney disease: unavailable/undetermined 50%, hypertension 21.1%, diabetes 18.9%, glomerulonephritis 6.7%, obstructive causes 1.2%, tubulointerstitial 0.9%, polycystic 0.4% and other 0.7%. Discussion: Despite the increase in RRT services, the prevalence is lower than the Latin American average (660 pmp). Three quarters of HD and PD patients are under-dialysed. Obsolete RRT techniques are still used. The presence of Mesoamerican nephropathy influences the demographic characteristics (many young patients, two-thirds male, high prevalence in lowlands and coastlands) (AU)

Prevalence of patients receiving renal replacement therapy in El Salvador in 2014. / Prevalencia de pacientes con tratamiento sustitutivo renal en El Salvador en 2014.

El Salvador has the highest renal failure mortality rate in the Americas. Five healthcare providers offer renal replacement therapy (RRT) in the country. The national RRT prevalence has never been reported. AIMS: To determine the RRT prevalence in El Salvador and some basic characteristics. METHODS: The association of nephrology coordinated a nationwide cross-sectional survey during the third quarter of 2014. 31 renal centres participated in the survey, covering 99.5% of patients. RESULTS: National RRT prevalence: 595 per million population (pmp), N=3807, average age 50.4 years, 67.5% male. By modality: peritoneal dialysis (PD) 289 pmp, haemodialysis (HD) 233 pmp, with functioning kidney transplantation 74 pmp (living donor only). Social security covers 25% of the population but treats 49.7% of RRT patients. Generally, higher prevalence was observed in municipalities with renal centres or located on the coast or lowlands. Ninety-five percent of HD patients receive fewer than 3 weekly sessions. Of PD patients, 59% do not belong to a continuous outpatient or automated programme, and 25% still use rigid catheter. Aetiology of chronic kidney disease: unavailable/undetermined 50%, hypertension 21.1%, diabetes 18.9%, glomerulonephritis 6.7%, obstructive causes 1.2%, tubulointerstitial 0.9%, polycystic 0.4% and other 0.7%. DISCUSSION: Despite the increase in RRT services, the prevalence is lower than the Latin American average (660 pmp). Three quarters of HD and PD patients are under-dialysed. Obsolete RRT techniques are still used. The presence of Mesoamerican nephropathy influences the demographic characteristics (many young patients, two-thirds male, high prevalence in lowlands and coastlands).

C.E.R.A. maintains stable hemoglobin in Latin American patients on dialysis.

BACKGROUND: C.E.R.A. is a continuous erythropoietin receptor activator with characteristics that permit a once-monthly schedule of administration for the maintenance treatment for chronic kidney disease (CKD) patients. The main objective of this study was to assess the maintenance of Hb concentration with once-monthly intravenous and/or subcutaneous C.E.R.A. therapy in Latin American dialysis patients with chronic renal anemia previously treated with epoetin alfa s.c or i.v 1-3 times per week. METHODS: This was a single-arm, open-label, multicenter, 32-week study of anemic patients with CKD previously treated with epoetin alfa sc or iv 1-3 times per week. After a 4-week screening period, during which mean Hb levels were maintained between 10.5 and 12.5 g/dL on their previous erythropoiesis stimulating agent, eligible patients entered a 16-week C.E.R.A. dose titration period followed by a 4-week efficacy evaluation period (EEP) and a 28-week safety follow-up. The starting dose of C.E.R.A. was based on the previous dose of epoetin alfa. Doses of C.E.R.A. were then adjusted to maintain Hb levels within ±1.0 g/dL of the reference concentration and between 10.5 and 12.5 g/dL. The Hb reference concentration was defined as the mean of all Hb levels during screening. The primary end point was the proportion of patients maintaining a mean Hb concentration (g/dL) within ±1 g/dL of their reference Hb and between 10.5 and 12.5 g/dL during the EEP. RESULTS: A total of 163 patients from 27 centers in Argentina, Brazil, Chile, Colombia, Ecuador, Mexico, Peru, Uruguay, and Venezuela entered the treatment period and 102 completed the prescribed course of C.E.R.A. Forty-five patients (43.7 %) maintained a mean Hb concentration within ±1 g/dL of their reference Hb value and between 10.5 and 12.5 g/dL during the EEP. The median monthly dose remained constant at 120 µg during the titration period and during the EEP. On the average, there were only 2.3 dose changes per patient in 28 weeks of treatment, covering 7 C.E.R.A. scheduled administrations. 53 % of all dose changes were dose decreases, 47 % increases. A total of 10 AEs and 4 SAEs were considered to be related to the study treatment. CONCLUSIONS: Once-monthly C.E.R.A. treatment effectively maintains stable Hb concentrations in patients with chronic renal anemia undergoing dialysis with a good safety and tolerability profile.

Increased resting intracellular calcium modulates NF-κB-dependent inducible nitric-oxide synthase gene expression in dystrophic mdx skeletal myotubes.

Duchenne muscular dystrophy (DMD) is a genetic disorder caused by dystrophin mutations, characterized by chronic inflammation and severe muscle wasting. Dystrophic muscles exhibit activated immune cell infiltrates, up-regulated inflammatory gene expression, and increased NF-κB activity, but the contribution of the skeletal muscle cell to this process has been unclear. The aim of this work was to study the pathways that contribute to the increased resting calcium ([Ca(2+)](rest)) observed in mdx myotubes and its possible link with up-regulation of NF-κB and pro-inflammatory gene expression in dystrophic muscle cells. [Ca(2+)](rest) was higher in mdx than in WT myotubes (308 ± 6 versus 113 ± 2 nm, p < 0.001). In mdx myotubes, both the inhibition of Ca(2+) entry (low Ca(2+) solution, Ca(2+)-free solution, and Gd(3+)) and blockade of either ryanodine receptors or inositol 1,4,5-trisphosphate receptors reduced [Ca(2+)](rest). Basal activity of NF-κB was significantly up-regulated in mdx versus WT myotubes. There was an increased transcriptional activity and p65 nuclear localization, which could be reversed when [Ca(2+)](rest) was reduced. Levels of mRNA for TNFα, IL-1ß, and IL-6 were similar in WT and mdx myotubes, whereas inducible nitric-oxide synthase (iNOS) expression was increased 5-fold. Reducing [Ca(2+)](rest) using different strategies reduced iNOS gene expression presumably as a result of decreased activation of NF-κB. We propose that NF-κB, modulated by increased [Ca(2+)](rest), is constitutively active in mdx myotubes, and this mechanism can account for iNOS overexpression and the increase in reactive nitrogen species that promote damage in dystrophic skeletal muscle cells.

El contexto en la epistemología personal: consideraciones teóricas y exploraciones empíricas / Context in personal epistemology: teoretical considerations and empirical explorations

En la investigación sobre epistemología personal, se ha tenido escasamente en cuenta el contexto. En este artículo, exponemos razones teóricas que muestran la importancia de considerar el contexto al que la epistemología personal hace referencia, así como el contexto en el que las creencias son declaradas. Proponemos que la comprensión epistemológica referida a y declarada en contextos especializados, podría ser diferente que la referida a y declarada en contextos no especializados. Presentamos dos estudios exploratorios cuyo objetivo fue determinar si la comprensión epistemológica es distinta en contextos referencial y declarativo distintos. Los resultados mostraron que la comprensión epistemológica referida a y declarada en el contexto pedagógico -especializado- es mayor que cuando es referida a y declarada en un contexto social informal - no especializado. Sugerimos que es importante incorporar los contextos declarativo y referencial, así como la distinción entre contexto especializado y no especializado, en la investigación en epistemología personal.(AU)

In personal epistemology research, context has been scarcely kept in mind. In this paper, theoretical reasons are presented that show the importance of considering the context to which the personal epistemology makes reference, as well as the context in which the beliefs are declared. We propose that epistemological understanding referred to and declared in specialized contexts, could be different that the one referred to and declared in non specialized contexts. Two exploratory studies were carried out in order to answer the research question: Are epistemological understanding different in different reference and declarative contexts? In both studies, epistemological understanding referred to and declared in the specialized pedagogic context was bigger than when it is referred to and declared in a non-specialized, an informal social context. It is suggested that it is important to incorporate declarative and referential contexts, as well as the distinction among specialized and non-specialized context, in personal epistemology research.(AU)

Parallel confocal detection of single molecules in real time.

The confocal detection principle is extended to a highly parallel optical system that continuously analyzes thousands of concurrent sample locations. This is achieved through the use of a holographic laser illumination multiplexer combined with a confocal pinhole array before a prism dispersive element used to provide spectroscopic information from each confocal volume. The system is demonstrated to detect and identify single fluorescent molecules from each of several thousand independent confocal volumes in real time.

Ulnar-basilic fistula: indications, surgical aspects, puncture technique, and results.

Ulnar-basilic fistula (UBF) is an autogenic vascular access option for hemodialysis (HD) first reported in 1967. However, only a few reports dealing with its complications and survival rates have been published in the literature. In the present work the results of 61 UBFs done in 60 adult patients on chronic HD are reported. Forty UBFs were created as primary access and the remaining 21 UBF as secondary access after: (a) thrombosis of an ipsilateral radiocephalic fistula (RCF) in 6 cases, (b) thrombosis of a contralateral RCF in 7 cases, and (c) thrombosis of a brachiocephalic fistula in 2 cases. No episodes of surgical complications, arterial steal, or ulnar nerve damage were observed. Five UBFs thrombosed within the first week after surgery. The 1-, 3-, and 5-year unassisted survival rates were 70.9%, 67.7%, and 57.3%, respectively, which were significantly higher than those reported previously. The inclusion of UBF in routine access plans is recommended.

Micosis superficial en poblaciones selváticas: prevalencia en cuatro comunidades rurales: Santa Rosa de Tiocayu, Pamashto, Bello Horizonte y Sachvacayoc / Superficial Micosis in wild populations: prevalencia in four rural communities: Santa Rosa of Tiocayu, Pamashto, Beautiful Horizon and Sachvacayoc

La micosis superficial constituye uno de los problemas dermatológicos más importantes, significando cuantiosas pérdidas económicas, desconociéndose en nuestro medio, una prevalencia real y factores asociados a dicha patología. Se estudio la prevalencia de micosis superficial en cuatro comunidades rurales del departamento de San Martín. Santa Rosa de Tioyacu, Pamashto y Bello Horizonte y en el departamento de Madre de Dios: Sachavacayoc, durante el mes de enero de 1999, a través de una encuesta y examen directo y cultivo. Los pacientes con lesiones clínicamente sospechosas de micosis superficial, ingresaron al estudio. La mayor prevalencia de Onicomicosis de mano, pitiriasis versicolor y tiña pedis corespondió a Bello Horizonte con 4.5 por ciento, 4.5 por ciento y 1.5 por ciento respectivamente. Para onicomicosis de pie, la mayor prevalencia correspondió a Santa Rosa de Tioyacu con 6.2 por ciento y para tiña corporis correspondió a Sachavacayoc con 9.5 por ciento. El dermatofito más frecuentemente aislado fue del género trichophyton y los cuadros clínicos más frecuentes fueron onicomicosis, tiña corporis y tiña pedis. En onicomicosis de mano, el género candida sp fue el más prevalente (83 por ciento) y la única especie de hongo aislado de tiña capitis en menores de 12 años fue trichophyton tonsurans. El examen directo ofreció una mayor proporción de hallazgos positivos 60.46 por ciento que el cultivo 6.97 por ciento y 37.20 por ciento de los casos dieron positividad de ambos métodos. Entre los factores asociados más frecuentes para tiña corporis fueron el contacto con algún miembro familiar infectado y contacto permanente con animales domésticos. En onicomicosis de pie, el uso de calzado hermético por más de 10 horas al día en promedio y actividades que implicaban contacto con agua por un tiempo prolongado como la agricultura y pesca fueron los más frecuentes. Se concluye que micosis superficial por dermatofitos es una patología dermatológica prevalente en comunidades rurales selváticas.

Evaluación de donantes de riñón 1 a 10 años después de nefrectomía / Kidney donors evaluation 1 to 10 years after nephrectomy

El presente reporte analiza los resultados obtenidos en la evaluación practicada a 56 donantes de riñón, controlados por períodos de 1 a 10 años postnefrectomía. La depeuración de creatinina (promedio ñ ES) fue de 87.0 ñ 4.05 ml/min, lo cual representa el 78% de los valores prenefrectomía y el 80.5% del de los controles. La creatinina sérica fue de 0.96 ñ 0.03 mg/dl. La incidencia de hipertensión arterial fue de 21.4% (p<0.001 vs controles), siendo más frecuente en los hombres. Proteinuria inferior a 500 mg/día se encontró en 3 de 56 donantes. En esta población de donantes la nefrectomía unilateral no significó un riesgo para la función renal del riñón remanente en los 10 años siguientes a la donación