Pharmacophore Identification and Structure-Activity Relationship Analysis of a Series of Substituted Azaindoles as Inhibitors of Trypanosoma brucei.

Human African trypanosomiasis is among the World Health Organization's designated neglected tropical diseases. Repurposing strategies are often employed in academic drug discovery programs due to financial limitations, and in this instance, we used human kinase inhibitor chemotypes to identify substituted 4-aminoazaindoles, exemplified by 1. Structure-activity and structure-property relationship analysis, informed by cheminformatics, identified 4s as a potent inhibitor of Trypanosoma brucei growth. While 4s appeared to be fast acting and cidal in the in vitro assays, it failed to cure a murine model of infection. Preliminary efforts to identify the potential mechanism of action of the series pointed to arginine kinase, though, as we demonstrate, this does not appear to be the sole target of our compounds. This comprehensive approach to drug discovery, encompassing cheminformatics, structure-potency and structure-property analysis, and pharmacophore identification, highlights our multipronged efforts to identify novel lead compounds for this deadly disease.

The Next Frontier in Health Disparities-A Closer Look at Exploring Sex Differences in Glioma Data and Omics Analysis, from Bench to Bedside and Back.

Sex differences are increasingly being explored and reported in oncology, and glioma is no exception. As potentially meaningful sex differences are uncovered, existing gender-derived disparities mirror data generated in retrospective and prospective trials, real-world large-scale data sets, and bench work involving animals and cell lines. The resulting disparities at the data level are wide-ranging, potentially resulting in both adverse outcomes and failure to identify and exploit therapeutic benefits. We set out to analyze the literature on women's data disparities in glioma by exploring the origins of data in this area to understand the representation of women in study samples and omics analyses. Given the current emphasis on inclusive study design and research, we wanted to explore if sex bias continues to exist in present-day data sets and how sex differences in data may impact conclusions derived from large-scale data sets, omics, biospecimen analysis, novel interventions, and standard of care management.

Gestión de riesgos y condicionalidad política de la cooperación internacional en tiempos de Coronavirus en Venezuela / Risk management and political conditionality of international cooperation in Venezuela during COVID-19

Este artículo examina la implementación en Venezuela del modelo de gestión de riesgo conocido como la Estrategia Internacional para la Reducción de Desastres de las Naciones Unidas (UNISDR,2015), o Marco de Sendai, ante la pandemia del COVID-19, así como la condicionalidad política por parte de donantes de Ayuda Oficial para el Desarrollo (AOD) en este contexto. Explora las decisiones políticas y medidas tomadas por el gobierno de la República Bolivariana de Venezuela, cuando apenas se confirmaban los primeros casos de contagio en la ciudad China de Wuhan, ratificados posteriormente el 3 de enero del 2020 por comunicado Oficial de la Organización Mundial de la Salud (OMS), a partir de lo cual se evalúa el ciclo de vida de la gestión del riesgo ante desastres, las políticas, decisiones y practicas adoptadas en Venezuela. Por otro lado, se aborda la intencionalidad política de la Cooperación Internacional, a través de la Ayuda Oficial para el Desarrollo (AOD), revelando patrones que inspiran el accionar de los donantes ante las necesidades y alineamientos ideológicos que asumen las naciones receptoras. Se concluye que la continua subordinación de los derechos humanos y la democracia a otras preocupaciones de política interior y exterior, en particular el interés económico, demuestra que ante la grave situación endémica que hoy atraviesa el planeta y con ello, la nación venezolana, las decisiones adoptadas en materia de AOD no solo socavan la credibilidad y la legitimidad de algunos gobernantes, sino que también limitan el impacto y la efectividad de la gestión de riesgos, ante lo que ya se puede catalogar como de desastre(AU)

This This article examines the implementation in Ven-ezuela of the risk management model known as the United Nations International Strategy for Disaster Reduction (UNIS-DR,2015), or Sendai Framework to the COVID-19 pandemic, as well as political conditionality by Official Development Aid (OAD) donors in this context. It explores the political decisions and measures taken by the government of the Bolivarian Republic of Venezuela, when the first cases of contagion were barely confirmed in the Chinese city of Wuhan, subsequently ratified on January 3rd, 2020, by Official Com-muniqué of the World Health Organization (WHO), from which life cycle of disaster risk management, policies, deci-sions and practices adopted in Venezuela, are assessed. On the other hand, the political intentionality of International Cooperation is addressed through the Official Development Aid (OAD), revealing patterns that inspire donor action in the face of the ideological needs and alignments that recipient nations assume. It is concluded that the continued subordination of human rights and democracy to other domestic and foreign policy concerns, in particular economic interest, make it clear that in the face of the serious endemic situation that is currently going through the planet as well as in the Venezuelan nation, decisions taken on OAD not only undermine the credibility and legitimacy of some leaders, but also limit the impact and effectiveness of risk management, in the face of what can already be classified as a disaster(AU)

Text segmentation ability predicts future reading efficiency in Spanish-speaking children.

Can the ability to parse unspaced texts (measured by a Text Segmentation Task, TST) index and predict reading efficiency in Spanish-speaking children? A sample of 1112 children (1st to 6th grade) was assessed. Additionally, two subsamples (51 children of 4th-5th grades and 71 children of 1st grade) were followed up. Our results indicate that the TST: a) reflects the acquisition of reading over primary school grades; b) reflects the teacher's judgment about the child's reading development; c) accurately predicts oral reading efficiency one and four years later year, in the former case even after removing the contributions of the IQ and oral reading speed. These results indicate that TST can be used to both index present -and predict future- reading achievements.

Hit-to-Lead Optimization of Benzoxazepinoindazoles As Human African Trypanosomiasis Therapeutics.

Human African trypanosomiasis (HAT) is a neglected tropical disease caused by infection with either of two subspecies of the parasite Trypanosoma brucei. Due to a lack of economic incentive to develop new drugs, current treatments have severe limitations in terms of safety, efficacy, and ease of administration. In an effort to develop new HAT therapeutics, we report the structure-activity relationships around T. brucei for a series of benzoxazepinoindazoles previously identified through a high-throughput screen of human kinase inhibitors, and the subsequent in vivo experiments for HAT. We identified compound 18, which showed an improved kinase selectivity profile and acceptable pharmacokinetic parameters, as a promising lead. Although treatment with 18 cured 60% of mice in a systemic model of HAT, the compound was unable to clear parasitemia in a CNS model of the disease. We also report the results of cross-screening these compounds against T. cruzi, L. donovani, and S. mansoni.

Conocimientos sobre salud oral en diabéticos atendidos en Atención Primaria / Knowledge about oral health in diabetic primary care patient

Objetivo: Determinar los conocimientos en salud oral en pacientes adultos con diabetes mellitus tipo 2, así como su relación con el nivel glucémico, factores sociodemográficos, hábitos en higiene oral y cuidados dentales profesionales. Diseño: Estudio observacional transversal. Emplazamiento: Consultas de medicina de familia de cuatro Centros de Salud. Participantes: Se seleccionaron mediante muestreo aleatorio accidental 247 pacientes diagnosticados de diabetes mellitus tipo 2. Mediciones Principales: Los pacientes contestaron un cuestionario sobre la relación entre salud oral y diabetes, clasificándolos en dos grupos de conocimientos en función de las respuestas correctas al cuestionario. También se registraron antecedentes sociales, autocuidados orales, uso de servicios dentales y los valores más recientes de hemoglobina glucosilada (HbA1c). Resultados: El grupo de pacientes sin estudios presentó un porcentaje de conocimientos bajos (CB) significativamente mayor (56,04 %) que el grupo de pacientes con estudios secundarios (32 %) y universitarios (16,67 %) (p<0,05). El porcentaje de pacientes con una mayor conciencia de la relación diabetes-salud oral fue mayor en la zona urbana (67,12 %) (p<0,05). La distribución de la frecuencia diaria de cepillado y el uso de pasta dental fluorada entre los dos grupos reflejó diferencias estadísticamente significativas (p<0,05). La variable HbA1c no reveló diferencias estadísticamente significativas en función del nivel de conocimientos. Conclusiones: Los conocimientos sobre salud oral de los pacientes con diabetes tipo 2 se relacionan con la frecuencia de cepillado, uso de complementos para higiene oral, lugar de residencia y nivel de estudios. El estudio mostró hábitos de higiene deficitarios y bajo uso de servicios sanitarios en relación con la salud oral

Objective: To determine the knowledge about oral health among adult patients with type 2 diabetes mellitus, as well as its relationship with glycemic level, sociodemographic factors, oral hygiene habits and professional dental care. Design: Observational cross-sectional study. Location: Primary Care consultations in four health centers. Participants: Accidental random sampling was used to include a total of 247 patients with type 2 diabetes mellitus. Main measures: Patients completed a questionnaire on the relationship between oral health and diabetes, classifying them into two knowledge groups according to the number of correct answers obtained. Data related to social background, oral self-care, use of dental services, and the most recent glycosylated hemoglobin (HbA1c) values were also collected. Results: Patients with little or no education showed a significantly higher percentage of limited knowledge (56.04%) than patients with secondary (32.14%) and university education (16.67%) (p<0.05). The percentage of patients with greater awareness of the link between diabetes and oral health was higher in urban areas (67.12%) (p<0.05). The distribution of daily toothbrushing frequency and the use of fluoride toothpaste showed statistically significant differences between the two groups (p<0.05). The variable HbA1c did not show statistically significant differences according to the level of knowledge. Conclusions: The knowledge about oral health of patients with type 2 diabetes is related to frequency of brushing, use of complements in oral hygiene, place of residence and educational level. The study also reflected deficiency in hygiene habits and in the use of healthcare services regarding oral health

Transgenerational Effects of pCO2-Driven Ocean Acidification on Adult Mussels Mytilus chilensis Modulate Physiological Response to Multiple Stressors in Larvae.

The effect of CO2-driven ocean acidification (OA) on marine biota has been extensively studied mostly on a single stage of the life cycle. However, the cumulative and population-level response to this global stressor may be biased due to transgenerational effects and their impacts on physiological plasticity. In this study, we exposed adult mussels Mytilus chilensis undergoing gametogenesis to two pCO2 levels (550 and 1200 µatm) for 16 weeks, aiming to understand if prolonged exposure of reproductive individuals to OA can affect the performance of their offspring, which, in turn, were reared under multiple stressors (pCO2, temperature, and dissolved cadmium). Our results indicate dependence between the level of pCO2 of the broodstock (i.e., parental effect) and the performance of larval stages in terms of growth and physiological rates, as a single effect of temperature. While main effects of pCO2 and cadmium were observed for larval growth and ingestion rates, respectively, the combined exposure to stressors had antagonistic effects. Moreover, we found a suppression of feeding activity in post-spawning broodstock upon high pCO2 conditions. Nevertheless, this observation was not reflected in the final weight of the broodstock and oocyte diameter. Due to the ecological and socioeconomic importance of mussels' species around the globe, the potential implications of maternal effects for the physiology, survival, and recruitment of larvae under combined global-change stressors warrant further investigation.

Técnica ahorradora de tejidos con injertos de piel parcial: sistema Meek versus sistema convencional mallado. Reporte de caso / Tissue sparing technique using partial skin transplantation: Meek technique versus conventional mesh technique. A case report

La técnica Meek es útil en pacientes con sitios donantes limitados, ya que es relativamente eficiente en cuanto a la expansión de la piel; llegando a expandirse hasta 9 veces en comparación al tamaño original, se realizó la evaluación de la cicatrización entre dos técnicas de expansión de tejidos de 1:3 veces entre la técnica Meek y la técnica mallado convencional. Cabe destacar que, con los avances de la bioingeniería, la técnica Meek se integra como un método completo para el tratamiento de autoinjertos. Presentamos un caso de un paciente varón de 34 años con una enfermedad infecciosa poli bacteriana "fascitis necrotizante"; con pérdida de tejidos que comprometen el tórax, abdomen y región inguino perineal, con un 14% de superficie corporal total comprometida. En la valoración cualitativa con la escala de Vancouver y POSAS se observaron mejores resultados de cicatrización con la técnica Meek.

The Meek technique is useful in patients with limited donor sites. It is a relatively efficient technique due to its ability to expand the skin up to nine times its original size. Scarring was assessed using two tissue expansion techniques, i.e. the Meek technique and the conventional mesh technique, which achieved an expansion ratio of 1:3. It should be noted that, with advances in bioengineering, the Meek technique is integrated as a complete method for autografting.1,2 We present the case of a 34-year-old male patient with a polymicrobial infectious disease called "necrotizing fasciitis". He had lost tissues of the thorax, abdomen and inguinal-perineal region, affecting 14% of his total body surface area. A qualitative assessment using the Vancouver scale and the Patient Objective Scar Assessment Scale (POSAS) showed better scarring results with the Meek technique

Accumulation and Biotransformation of Dinophysis Toxins by the Surf Clam Mesodesma donacium.

Surf clams, Mesodesma donacium, were shown to accumulate toxins from Dinophysis acuminata blooms. Only pectenotoxin 2 (PTX2) and some of its derivatives were found, and no toxins from the okadaic acid group were detected. PTX2 seems to be transformed to PTX2 seco-acid (PTX2sa), which was found in concentrations more than ten-fold those of PTX2. The seco-acid was transformed to acyl-derivatives by esterification with different fatty acids. The estimated amount of these derivatives in the mollusks was much higher than that of PTX2. Most esters were originated by even carbon chain fatty acids, but some originated by odd carbon number were also found in noticeable concentrations. Some peaks of toxin in the bivalves did not coincide with those of Dinophysis abundance, suggesting that there were large differences in toxin content per cell among the populations that developed throughout the year. The observed depuration (from the digestive gland) was fast (more than 0.2 day-1), and was faster for PTX2 than for PTX2sa, which in turn was faster than that of esters of PTX2sa. PTX2 and PTX2sa were distributed nearly equally between the digestive gland and the remaining tissues, but less than 5% of the palmytoyl-esters were found outside the digestive gland.

Batería de Rendimiento Intelectual para niños escolares cubanos / Battery intellectual performance for Cuban children

En Cuba, no existen instrumentos propios para la evaluación de la inteligencia. Para solucionar el problema, el Centro de Neurociencias de Cuba, y en especial, el Departamento de Neurociencias Educacionales, diseñó una Batería de Rendimiento Intelectual (BRI) para niños escolares cubanos (conformado por Escala capacidad Intelectual, ECI y Analogías verbales, AV). Objetivos: (1) Completar y validar la BRI para la evaluación de Gf y Gc en niños cubanos de edad escolar, (2) Método. Determinar la contribución única de Gf y Gc, evaluada con la BRI, en el aprovechamiento académico en niños cubanos. Se aplicaron dos estudios. a) Validación de la ECI con una muestra de 370 niños entre 7 y 11 años (N1 ­ 270 para calcular punto de corte y N2 para validar los puntos de corte). b) Diseño de la AV con una muestra de 36 niños entre 10 y 11 años. Resultados. La ECI resulta una prueba apropiada para detectar al niño escolar con riesgo intelectual. El resultado se sustenta en la alta coincidencia (89%) con el WISC-R para clasificar niños con trastornos intelectuales. El diseño de la AV presenta apropiadas propiedades psicométricas, pues presenta significativas correlaciones con la sub-prueba de Vocabulario del WISC-R. Conclusiones. La BRI presenta apropiadas propiedades psicométricas para detectar el riesgo intelectual en niños cubanos.

In Cuba not have own instruments for the assessment of intelligence. To resolve the problem, the Neuroscience Center of Cuba, and the Department of Educational Neuroscience, design a Battery of Intellectual Performance (BIP) for school children (Intellectual Capacity Scale, ICE and Verbal Analogy, VA). Objectives: (1) to complete and validate the BIP for the evaluation of Gf and Gc in cuban children of school age, (2) Method. Determine the unique contribution of Gf and Gc, evaluated with the BIP, on academic achievement in Cuban children. Two studies were applied. a) Validation of the ICE with a sample of 370 children between 7 and 11 years (N1 - 270 to calculate the cut-off point and N2 to validate the cut-off points). b) Design of the VA with a sample of 36 children between the ages of 10 and 11 years. Results. The ICE is a proper test to detect the child with intellectual risk. The result is based on the high coincidence (89%) with the WISC-R to classify children with intellectual disorders. The design of the VA presents appropriate psychometric properties, because it presents significant correlations with the sub-vocabulary test of the WISC-R. Conclusions. The BIP presents appropriate psychometric properties to detect risk Cuban intellectual in school children.

Contribución única de la inteligencia fluida y cristalizada en el rendimiento académico / Unique contribution of intelligence fluid and crystallized from academic performance

La teoría de la inversión de Cattell establece que inteligencia fluida(Gf) constituye la base de la inteligencia cristalizada (Gc), en tanto hace posible la adquisición de habilidades y conocimientos. Sin embargo, las evidencias empíricas no son concluyentes. La presente investigación pretende determinar la contribución única de la inteligencia, Gf y Gc, sobre el rendimiento académico del niño escolar. Se utilizó una muestra de 100 niños según criterios de selección en dos tipos de enseñanza: niños con calificaciones de Mal y Regular en las materias escolares (enseñanza regular) y niños con retraso mental ligero (enseñanza especial). Las pruebas intelectuales fueron: Escala de Capacidad Intelectual y Analogía verbales. Las tareas académicas fueron: fluidez lectora y fluidez matemática. Los resultaron indicaron fuertes correlaciones entre las variables académicas y las intelectuales. La regresión jerárqica indicó contribución significativa de Gc (Analogía verbales) para pronosticar el desarrollo de la lectura cuando se controla el efecto de Gf (Escala de Capacidad Intelectual). Por el contrario, Gf mostró una contribución significativa para explicar el rendimiento de la matemática cuando se controla a Gc.

The investment theory of Cattell states that fluid intelligence (Gf) is the basis of crystallized intelligence (Gc), as does the acquisition of skills and knowledge possible. However, the empirical evidence is inconclusive. This research trying to determine the unique contribution of intelligence, Gf and Gc, on the academic performance of school children. A sample of 100 children was used as selection criteria in two types of education: children with scores bad and regular school subjects (regular education) and children with mild mental retardation (special education). Intellectual tests were: Intellectual Ability Scale and Verbal Analogy test. Academic tasks were: reading fluency and math fluency. The resulted showed strong correlations between academic and intellectual variables. The hierarchical regression indicated significant contribution of Gc (Verbal Analogy) to predict the development of reading when the effect of Gf (Intellectual Ability Scale) is controlled. By contrast, Gf showed a significant contribution to explain the performance of mathematics when Gc controls.

Identification of "Preferred" Human Kinase Inhibitors for Sleeping Sickness Lead Discovery. Are Some Kinases Better than Others for Inhibitor Repurposing?

A kinase-targeting cell-based high-throughput screen (HTS) against Trypanosoma brucei was recently reported, and this screening set included the Published Kinase Inhibitor Set (PKIS). From the PKIS was identified 53 compounds with pEC50 ≥ 6. Utilizing the published data available for the PKIS, a statistical analysis of these active antiparasitic compounds was performed, allowing identification of a set of human kinases having inhibitors that show a high likelihood for blocking T. brucei cellular proliferation in vitro. This observation was confirmed by testing other established inhibitors of these human kinases and by mining past screening campaigns at GlaxoSmithKline. Overall, although the parasite targets of action are not known, inhibitors of this set of human kinases displayed an enhanced hit rate relative to a random kinase-targeting HTS campaign, suggesting that repurposing efforts should focus primarily on inhibitors of these specific human kinases. We therefore term this statistical analysis-driven approach "preferred lead repurposing".

A Replicative In Vitro Assay for Drug Discovery against Leishmania donovani.

The protozoan parasite Leishmania donovani is the causative agent of visceral leishmaniasis, a disease potentially fatal if not treated. Current available treatments have major limitations, and new and safer drugs are urgently needed. In recent years, advances in high-throughput screening technologies have enabled the screening of millions of compounds to identify new antileishmanial agents. However, most of the compounds identified in vitro did not translate their activities when tested in in vivo models, highlighting the need to develop more predictive in vitro assays. In the present work, we describe the development of a robust replicative, high-content, in vitro intracellular L. donovani assay. Horse serum was included in the assay media to replace standard fetal bovine serum, to completely eliminate the extracellular parasites derived from the infection process. A novel phenotypic in vitro infection model has been developed, complemented with the identification of the proliferation of intracellular amastigotes measured by EdU incorporation. In vitro and in vivo results for miltefosine, amphotericin B, and the selected compound 1 have been included to validate the assay.

Identification and characterization of hundreds of potent and selective inhibitors of Trypanosoma brucei growth from a kinase-targeted library screening campaign.

In the interest of identification of new kinase-targeting chemotypes for target and pathway analysis and drug discovery in Trypanosomal brucei, a high-throughput screen of 42,444 focused inhibitors from the GlaxoSmithKline screening collection was performed against parasite cell cultures and counter-screened against human hepatocarcinoma (HepG2) cells. In this way, we have identified 797 sub-micromolar inhibitors of T. brucei growth that are at least 100-fold selective over HepG2 cells. Importantly, 242 of these hit compounds acted rapidly in inhibiting cellular growth, 137 showed rapid cidality. A variety of in silico and in vitro physicochemical and drug metabolism properties were assessed, and human kinase selectivity data were obtained, and, based on these data, we prioritized three compounds for pharmacokinetic assessment and demonstrated parasitological cure of a murine bloodstream infection of T. brucei rhodesiense with one of these compounds (NEU-1053). This work represents a successful implementation of a unique industrial-academic collaboration model aimed at identification of high quality inhibitors that will provide the parasitology community with chemical matter that can be utilized to develop kinase-targeting tool compounds. Furthermore these results are expected to provide rich starting points for discovery of kinase-targeting tool compounds for T. brucei, and new HAT therapeutics discovery programs.

Establishment of a structure-activity relationship of 1H-imidazo[4,5-c]quinoline-based kinase inhibitor NVP-BEZ235 as a lead for African sleeping sickness.

Compound NVP-BEZ235 (1) is a potent inhibitor of human phospoinositide-3-kinases and mammalian target of rapamycin (mTOR) that also showed high inhibitory potency against Trypanosoma brucei cultures. With an eye toward using 1 as a starting point for anti-trypanosomal drug discovery, we report efforts to reduce host cell toxicity, to improve the physicochemical properties, and to improve the selectivity profile over human kinases. In this work, we have developed structure-activity relationships for analogues of 1 and have prepared analogues of 1 with improved solubility properties and good predicted central nervous system exposure. In this way, we have identified 4e, 9, 16e, and 16g as the most promising leads to date. We also report cell phenotype and phospholipidomic studies that suggest that these compounds exert their anti-trypanosomal effects, at least in part, by inhibition of lipid kinases.

Third target of rapamycin complex negatively regulates development of quiescence in Trypanosoma brucei.

African trypanosomes are protozoan parasites transmitted by a tsetse fly vector to a mammalian host. The life cycle includes highly proliferative forms and quiescent forms, the latter being adapted to host transmission. The signaling pathways controlling the developmental switch between the two forms remain unknown. Trypanosoma brucei contains two target of rapamycin (TOR) kinases, TbTOR1 and TbTOR2, and two TOR complexes, TbTORC1 and TbTORC2. Surprisingly, two additional TOR kinases are encoded in the T. brucei genome. We report that TbTOR4 associates with an Armadillo domain-containing protein (TbArmtor), a major vault protein, and LST8 to form a unique TOR complex, TbTORC4. Depletion of TbTOR4 caused irreversible differentiation of the parasite into the quiescent form. AMP and hydrolysable analogs of cAMP inhibited TbTOR4 expression and induced the stumpy quiescent form. Our results reveal unexpected complexity in TOR signaling and show that TbTORC4 negatively regulates differentiation of the proliferative form into the quiescent form.

Declaración de voluntades anticipadas: estudio cualitativo en personas mayores y médicos de Atención Primaria / Living will declarations: Qualitative study of the elderly and primary care general practioners

ObjetivosEn primer lugar, evaluar los conocimientos y actitudes de los mayores de 65 años respecto al documento de voluntades anticipadas, y conocer el papel que, en su opinión, debe jugar el médico de familia en la planificación anticipada de la muerte. En segundo lugar, evaluar los conocimientos y actitudes de los médicos de familia respecto a dicho documento y conocer el papel que juegan estos profesionales.DiseñoCualitativo, utilizando grupos de discusión. Se realizaron 4 grupos, 2 de médicos y 2 de mayores entre marzo y mayo de 2009.EmplazamientoAtención Primaria de Castilla-La Mancha, Áreas de Salud de Albacete y Guadalajara.ParticipantesQuince personas en los grupos de mayores y 13 en los de médicos.MétodoTanto para la selección de sujetos, la realización de los grupos y el análisis de estos, se siguieron las recomendaciones de Krueger (1991).ResultadosLos médicos y las personas mayores presentan una actitud positiva hacia el documento de voluntades anticipadas, pero carecen de suficiente información para que su uso pueda ser más extendido. El desconocimiento hace imposible que los pacientes reclamen información y la falta de tiempo dificulta que los médicos planteen el tema en la consulta.ConclusionesTanto los médicos como los mayores no disponen de información suficiente sobre el documento de voluntades anticipadas, lo cual permite recomendar la realización de campañas informativas y/o actividades de formación específicas. Los médicos creen que podrían buscarse alternativas a las consultas de Atención Primaria para que los pacientes pudieran informarse(AU)

ObjectivesFirst, to assess knowledge and attitudes of people aged 65 and over to the Advance Directives Document (ADD) and know the role that, in their opinion, a family doctor should play in the early planning of death. Second, to assess the level of knowledge and attitude of primary care physicians to that document, as well as to know the role played by these professionals.DesignQualitative, using discussion groups. Between March and May 2009 4 groups, 2 of physicians and 2 of older people, were formed.SettingCastile-La Mancha Primary Care Setting, Albacete and Guadalajara health areas.Subjects15 people in the groups formed by older people and 13 in the groups of doctors.MethodThe Krueger recommendations (1991) were followed both for the selection of subjects, conducting of the discussion groups and analysis of data.ResultsDoctors and patients have a positive attitude towards the ADD, but lack sufficient information to make more widespread use of it. Ignorance makes it impossible for patients to get information about the document and the lack of time makes it difficult for physicians to raise the issue in the consultation.ConclusionsNeither physicians nor the elderly have enough information regarding ADD, allowing us to recommend carrying out information campaigns and/or specific educational activities. According to physicians, alternatives to primary care consultations should be sought for patients to be informed(AU)

[Living will declarations: Qualitative study of the elderly and primary care general practitioners]. / Declaración de voluntades anticipadas: estudio cualitativo en personas mayores y médicos de Atención Primaria.

OBJECTIVES: First, to assess knowledge and attitudes of people aged 65 and over to the Advance Directives Document (ADD) and know the role that, in their opinion, a family doctor should play in the early planning of death. Second, to assess the level of knowledge and attitude of primary care physicians to that document, as well as to know the role played by these professionals. DESIGN: Qualitative, using discussion groups. Between March and May 2009 4 groups, 2 of physicians and 2 of older people, were formed. SETTING: Castile-La Mancha Primary Care Setting, Albacete and Guadalajara health areas. SUBJECTS: 15 people in the groups formed by older people and 13 in the groups of doctors. METHOD: The Krueger recommendations (1991) were followed both for the selection of subjects, conducting of the discussion groups and analysis of data. RESULTS: Doctors and patients have a positive attitude towards the ADD, but lack sufficient information to make more widespread use of it. Ignorance makes it impossible for patients to get information about the document and the lack of time makes it difficult for physicians to raise the issue in the consultation. CONCLUSIONS: Neither physicians nor the elderly have enough information regarding ADD, allowing us to recommend carrying out information campaigns and/or specific educational activities. According to physicians, alternatives to primary care consultations should be sought for patients to be informed.

Biochemical and molecular characterization of Coriolopsis rigida laccases involved in transformation of the solid waste from olive oil production.

Two laccase isoenzymes were purified and characterized from the basidiomycete Coriolopsis rigida during transformation of the water-soluble fraction of "alpeorujo" (WSFA), a solid residue derived from the olive oil production containing high levels of toxic compounds. Zymogram assays of laccases secreted by the fungus growing on WSFA and WSFA supplemented with glucose showed two bands with isoelectric points of 3.3 and 3.4. The kinetic studies of the two purified isoenzymes showed similar affinity on 2,6-dimethoxyphenol and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid), used as phenolic and non-phenolic model substrate, respectively. The molecular mass of both proteins was 66 kDa with 9% N-linked carbohydrate. Physico-chemical properties of the purified laccases from media containing WSFA were similar to those obtained from medium with glucose as the main carbon source. In-vitro studies performed with the purified laccases revealed a 42% phenol reduction of WSFA, as well as changes in the molecular mass distribution. These findings indicate that these laccases are involved in the process of transformation, via polymerization by the oxidation of phenolic compounds present in WSFA. A single laccase gene, containing an open reading frame of 1,488 bp, was obtained in PCR amplifications performed with cDNA extracted from mycelia grown on WSFA. The product of the gene shares 90% identity (95% similarity) with a laccase from Trametes trogii and 89% identity (95% similarity) with a laccase from Coriolopsis gallica. This is the first report on purification and molecular characterization of laccases directly involved in the transformation of olive oil residues.