Modular Fluorescent Cholesterol Naphthalimide Probes And Their Application For Cholesterol Trafficking Studies In Cells.

Development of fluorescent cholesterol analogs to better understand subcellular cholesterol trafficking is of great interest for cell biology and medicine. Our approach utilizes a bifunctional 1,8-naphthalimide scaffold with a push-pull character, modified on one side with a head group and a linker on the other side connecting it to cholesterol via an ester bond. Through structure-function studies, we've explored how different substituents-linkers and head groups-affect the ability of these fluorescent cholesterol naphthalimide analogs (CNDs) to mimic natural cholesterol behavior at both molecular and cellular levels. We categorized the resulting analogs into three groups: neutral, charged, and those featuring a hydroxyl group. Each compound was assessed for its solvatochromic behavior in organic solvents and model membranes. Extensive all-atom molecular dynamics simulations helped us examine how these analogs perform in model membranes compared to cholesterol. Additionally, we investigated the partitioning of these fluorescent probes in phase-separated giant unilamellar vesicles. We evaluated the uptake and distribution of these probes within mouse fibroblast cells and astrocytes, for their subcellular distributions in lysosomes and compared that to BODIPY-cholesterol, a well-regarded fluorescent cholesterol analog. The internalization efficiency of the fluorescent probes varies in different cell types and is affected mainly by the head groups. Our results demonstrate that the modular design significantly simplifies the creation of fluorescent cholesterol probes bearing distinct spectral, biophysical, and cellular targeting features, which makes it a valuable toolkit for the investigation of subcellular distribution and trafficking of cholesterol and its derivatives.

Use of pure recombinant human enzymes to assess the disease-causing potential of missense mutations in urea cycle disorders, applied to N-acetylglutamate synthase deficiency.

N-acetylglutamate synthase (NAGS) makes acetylglutamate, the essential activator of the first, regulatory enzyme of the urea cycle, carbamoyl phosphate synthetase 1 (CPS1). NAGS deficiency (NAGSD) and CPS1 deficiency (CPS1D) present identical phenotypes. However, they must be distinguished, because NAGSD is cured by substitutive therapy with the N-acetyl-L-glutamate analogue N-carbamyl-L-glutamate, while curative therapy of CPS1D requires liver transplantation. Since their differentiation is done genetically, it is important to ascertain the disease-causing potential of CPS1 and NAGS genetic variants. With this goal, we previously carried out site-directed mutagenesis studies with pure recombinant human CPS1. We could not do the same with human NAGS (HuNAGS) because of enzyme instability, leading to our prior utilization of a bacterial NAGS as an imperfect surrogate of HuNAGS. We now use genuine HuNAGS, stabilized as a chimera of its conserved domain (cHuNAGS) with the maltose binding protein (MBP), and produced in Escherichia coli. MBP-cHuNAGS linker cleavage allowed assessment of the enzymatic properties and thermal stability of cHuNAGS, either wild-type or hosting each one of 23 nonsynonymous single-base changes found in NAGSD patients. For all but one change, disease causation was accounted by the enzymatic alterations identified, including, depending on the variant, loss of arginine activation, increased Km Glutamate, active site inactivation, decreased thermal stability, and protein misfolding. Our present approach outperforms experimental in vitro use of bacterial NAGS or in silico utilization of prediction servers (including AlphaMissense), illustrating with HuNAGS the value for UCDs of using recombinant enzymes for assessing disease-causation and molecular pathogenesis, and for therapeutic guidance.

The E3 ubiquitin ligase COP1 regulates salt tolerance via GIGANTEA degradation in roots.

Excess soil salinity significantly impairs plant growth and development. Our previous reports demonstrated that the core circadian clock oscillator GIGANTEA (GI) negatively regulates salt stress tolerance by sequestering the SALT OVERLY SENSITIVE (SOS) 2 kinase, an essential component of the SOS pathway. Salt stress induces calcium-dependent cytoplasmic GI degradation, resulting in activation of the SOS pathway; however, the precise molecular mechanism governing GI degradation during salt stress remains enigmatic. Here, we demonstrate that salt-induced calcium signals promote the cytoplasmic partitioning of CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1), leading to the 26S proteasome-dependent degradation of GI exclusively in the roots. Salt stress-induced calcium signals accelerate the cytoplasmic localization of COP1 in the root cells, which targets GI for 26S proteasomal degradation. Align with this, the interaction between COP1 and GI is only observed in the roots, not the shoots, under salt-stress conditions. Notably, the gi-201 cop1-4 double mutant shows an enhanced tolerance to salt stress similar to gi-201, indicating that GI is epistatic to COP1 under salt-stress conditions. Taken together, our study provides critical insights into the molecular mechanisms governing the COP1-mediated proteasomal degradation of GI for salt stress tolerance, raising new possibilities for developing salt-tolerant crops.

Molecular and Serological Studies on Potential SARS-CoV-2 Infection among 43 Lemurs under Human Care-Evidence for Past Infection in at Least One Individual.

In the setting of the recent COVID-19 pandemic, transmission of SARS-CoV-2 to animals has been reported in both domestic and wild animals and is a matter of concern. Given the genetic and functional similarities to humans, non-human primates merit particular attention. In the case of lemurs, generally considered endangered, they are believed to be susceptible to SARS-CoV-2 infection. We have conducted a study for evidence of SARS-CoV-2 infection among the 43 lemurs of Mundomar, a zoological park in Benidorm, Spain. They belong to two endangered lemur species, 23 black-and-white ruffed lemurs (Varecia variegata) and 20 ring-tailed lemurs (Lemur catta). Health assessments conducted in 2022 and 2023 included molecular analyses for SARS-CoV-2 RNA of oral and rectal swabs using two different RT-qPCR assays, always with negative results for SARS-CoV-2 in all animals. The assessment also included serological testing for antibodies against the receptor-binding domain (RBD) of the spike protein (S) of SARS-CoV-2, which again yielded negative results in all animals except one black-and-white ruffed lemur, supporting prior infection of that animal with SARS-CoV-2. Our data, while not indicating a high susceptibility of lemurs to SARS-CoV-2 infection, show that they can be infected, adding to the existing information body on potential ways for SARS-CoV-2 virus spreading in zoos, highlighting the need for animal surveillance for the virus.

Use of micafungin as antifungal prophylaxis in patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) in Spain (GETH-MIC) / Profilaxis antifúngica con micafungina en pacientes que reciben un trasplante alogénico de progenitores hematopoyéticos (alo-TPH) en España (GETH-MIC)

INTRODUCTION: The fungal infections remain an important problem in the allogeneic stem cell trasnsplantation (allo-SCT) setting and thus, anti-fungal prophylaxis is commonly used. The antifungal drug should offer activity, at least against Candida and Aspergillus spp., a good safety profile and low probability interactions. Micafungin could theoretically fulfill these requisites. The aim of the study was to describe the experience with micafungin as primary prophylaxis in patients undergoing allo-SCT in a cohort of Spanish centres, and to evaluate its efficacy and tolerability in this population. MATERIAL AND METHODS: Retrospective multicentre observational study including all consecutive adult patients admitted for allo-SCT in participating centres of the Grupo Español de Trasplante Hematopoyético (GETH), from January 2010 to December 2013, who received micafungin as primary prophylaxis during the neutropenic period. RESULTS: A total of 240 patients from 13 centres were identified and 159 patients were included for the analysis. Most patients (95.6%) received 50 mg/day of micafungin. During the follow-up, 7 (4.4%) patients developed breakthrough invasive fungal disease, 1 proven and 6 probable; one patient discontinued the drug because of serious drug interactions. Prophylaxis with micafungin was considered effective in 151 (94.9%) patiens. CONCLUSIONS: According to our experience, micafungin is an appropriate alternative for antifungal prophylaxis in patients undergoing an allo-HSCT, because its efficacy, its low profile of drug interactions and side-effects

INTRODUCCIÓN: Las infecciones fúngicas siguen representando un problema en el trasplante alogénico de progenitores hematopoyéticos (alo-TPH) por lo que es habitual el uso de profilaxis antifúngica en estos PACIENTES: El tratamiento antifúngico debe presentar al menos actividad frente a Candida y Aspergillus spp, un buen perfil de seguridad y baja probabilidad de infecciones, siendo micafungina una de las opciones que podría cumplir todos estos requisitos. El objetivo del estudio fue describir la experiencia con micafungina como profilaxis primaria en pacientes sometidos a alo-TPH en una cohorte de hospitales españoles, y evaluar su eficacia y seguridad en esta población. MATERIAL Y MÉTODOS: Estudio retrospectivo multicéntrico observacional consecutivo de todos los pacientes adultos ingresados para alo-TPH en los centros del Grupo Español de Trasplante Hematopoyético (GETH) desde enero de 2010 a diciembre de 2013 y que recibieron micafungina como profilaxis primaria durante el periodo de neutropenia. RESULTADOS: Se identificaron 240 pacientes de 13 hospitales y 159 fueron incluidos para el análisis. La mayoría (95.6%) de ellos recibieron dosis de 50mg/día de micafungina. Durante el seguimiento, 7 (4.4%) pacientes desarrollaron infecciones de brecha, 1 probada y 6 probables; en un paciente se suspendió el tratamiento por interacciones medicamentosas graves. La profilaxis con micafungina se consideró efectiva en el 94,9% de los pacientes (151 de 159). CONCLUSIONES: En base a nuestros resultados, consideramos que micafungina es una buena alternativa como profilaxis antifúngica en pacientes sometidos a alo-TPH, por su eficacia, el bajo riesgo de interacciones y de efectos adversos

Mortalidad innecesariamente prematura y sanitariamente evitable en Costa Rica / Unnecessary premature and avoidable mortality in Costa Rica

Fundamentos: Para desarrollar políticas y estrategias orientadas a mitigarlas es fundamental identificar las desigualdades. El objetivo del trabajo es analizar las variaciones de la Mortalidad Innecesariamente Prematura y Sanitariamente Evitable (MIPSE) para cada uno de los 81 cantones de Costa Rica durante el período 2000-2005. Métodos: Se aplicó la clasificación MIPSE propuesta por miembros del Servicio de Información y Estudios de la Dirección General de Recursos Sanitarios de Catalunya. Mediante el empleo de el Indicador de Desarrollo Socioeconómico (IDSE) establecido por economistas de la Universidad de Costa Rica, previa estandarización de la población, se ordenaron los cantones en grupos por quintiles (I el más rico, V el más pobre). Resultados: Se encontraron como causas principales de mortalidad MIPSE la enfermedad isquémica del corazón (19,55% causas MIPSE), accidentes de tránsito con vehículos a motor (11,60%), enfermedades cerebrovasculares (6,95%), perinatal (6,92%) y suicidios (5,14%). Conclusión: La mortalidad por HIV y el Sida, el cáncer de mamá en mujeres, cáncer de cuerpo de útero, cáncer de piel y por hepatitis secundaria al consumo de alcohol, afectan más a los cantones con mayores ingresos. La mortalidad por hiperplasia benigna de próstata, la materna asociada al embarazo, parto o puerperio y la hernia abdominal afectan más a los de menor nivel económico. Se identificaron dos grupos de MIPSE con desigualdad equidistribuida: leucemia y enfermedades cardiovasculares congénitas(AU)

Backgrounds: With the intention of establishing economic inequities, the article analyzes the variations of the Unnecessarily Premature and Sanitarily Avoidable Mortality (MIPSE) of each of the 81 cantons of Costa Rica during 2000-2005. It is important to identify those inequities, to establish policies and strategies trying to mitigate them. Method: It applies the MIPSE classification proposed by members of the Information and Studies Service, of the Catalunya’s Sanitary Resources Headquarter, Spain. By an Indicator of Socioeconomic Development (IDSE) of a University of Costa Rica economist’s team, it organised each canton in groups of quintiles (I for the richest, V for the poorest), previous people standardization. Results: We found as a major causes of mortality MIPSE in the country: Heart Isquemic Disease (19,55% MIPSE causes), Traffic Accidents with Motor Vehicles (11,60%), Brain Vascular Disease (6,95%), Perinatal (6,92%) and Suicide (5,14%). Conclusion: The VIH infection - AIDS mortality, the Best Cancer in Women, Uterus Cancer, Skin Cancer and Hepatic Disease Secondary to Alcohol Consumption, affects more the cantons with better financial conditions and the Prostate Benign Hyperplasia mortality, Mothers mortality related with Pregnancy, Childbirth or Puerperal Stage and the Abdominal Hernia mortality, affects more to those with worst economic level. Two MIPSE groups were identified with similar inequality: Leukaemia and Congenital Cardiovascular Disease(AU)

Los incentivos para la mejora de la calidad en los servicios de salud / Incentives for quality improvement in health services

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Impacto de la economía en la política y gestión sanitaria / The impact of economics on health policy and management in Spain

Fundamento: Las disciplinas científicas, como la Economía,tienen valor per se, conviene, sin embargo, aproximar el impacto desus aplicaciones en el bienestar social, o como mínimo –caso de laEconomía de la Salud (ES)– en la política y gestión sanitarias.Métodos: Se atiende a las tres vertientes relevantes del conocimiento(producción de nuevo conocimiento, difusión y aplicación)utilizando más la perspectiva 'emic' –la empleada en antropologíabasándose en la experiencia de los integrantes de una cultura–, que laperspectiva 'etic' asentada en descripciones materiales y dudosasestadísticas.Resultados: La solidez de los principios y resultados de la ESdepende de la base disciplinaria en la que se apoya, mientras que surelevancia –que no su traslación a la práctica– está vinculada a la delas cuestiones a las que atiende. Se registran las aportaciones relevantesde la Economía al ámbito sanitario. El grado de desarrollo dela ES en España resulta notable: séptima posición mundial pese a losrelativamente menores contenidos de ES en las revistas clínicas y deinvestigación sobre servicios sanitarios de España. La ES tiene enEspaña más presencia que influencia, no habiendo logrado impregnarsuficientemente la práctica diaria.Conclusiones: Los conocimientos de Economía que necesita unpolítico o un gestor sanitario o un clínico son limitados; el impactode la ES pasará, sobre todo, por educarles el olfato

Background: Despite the intrinsic value of scientific disciplines,such as Economics, it is appropriate to gauge the impact of itsapplications on social welfare, or at least –Health Economics' (HE)case– its influence on health policy and management.Methods: The three relevant features of knowledge (production,diffusion and application) are analyzed, more from an 'emic' perspective–the one used in Anthropology relying on the experience ofthe members of a culture– than from an 'etic' approach seated onmaterial descriptions and dubious statistics.Results: The soundness of the principles and results of HEdepends on its disciplinary foundations, whereas its relevance –thandoes not imply translation into practice– is more linked with the problemsstudied. Important contributions from Economics to the healthsphere are recorded. HE in Spain ranks seventh in the world despitethe relatively minor HE contents of its clinical and health servicesresearch journals. HE has in Spain more presence than influence,having failed to impregnate sufficiently the daily events.Conclusions: HE knowledge required by a politician, a healthmanager or a clinician is rather limited; the main impact of HE couldbe to develop their intuition and awareness

Comercio y salud / Trade and health

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Urgencias hospitalarias o colapso crónico: los pacientes crónicos no deberían colapsar urgencias / Chronic saturation of emergency departments: they should not be flooded by patients with chronic diseases

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Mejor (servicio) siempre es más (salud) / Better (Care) is always more (Health)

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Happy microbes in hostile niches. A symposium on extremophiles

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Noticias de El Cairo: 18ª Conferencia Internacional sobre Arginina y Pirimidinas / News from Cairo: 18th International Conference on Arginine and Pyrimidines

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Sistemas de ajuste por riesgo / Risk adjustment systems

El modelo de reembolso capitativo ajustado según el nivel de riesgo de los beneficios de un servicio de salud busca, por un lado, promover conductas eficientes al aproximar el pago a los gastos realmente justificados, y por otro mejorar el acceso de los usuarios de mayor riesgo a los servicios que les sean necesarios evitando la selección de pacientes que puede producirse ante sistemas captativos simples (sin ajuste). El objetivo de esta revisión es, además de describir el contexto en el que surge esta alternativa de reembolso -separación de las funciones de financiación, compra y provisión en los sistemas de salud-, contribuir en el análisis del alcance, ventajas y limitaciones, que a punto de partida de la evidencia disponible puede destacarse de la aplicación de los sistemas de ajuste por riesgo. La mayor parte de los mecanismos actuales de ajuste explícito de riesgo consideran las variables demográficas, las que en general obtienen una baja predicción del gasto. Se han incorporado otras dimensiones como la percepción del estado de salud por parte de los individuos, utilización previa de los servicios, y la tendencia más reciente, la información diagnóstica o la casuística de los pacientes. Cada una de estas alternativas tiene ventajas e inconvenientes, pero globalmente puede afirmarse que la varianza total explicada por los distintos modelos a un nivel individual aparece como muy pequeña. De esa pequeña proporción explicable, los modelos que obtienen mejores resultados son aquellos que incorporan información diagnóstica. Luego de revisar algunos de los requisitos que se recomiendan para la implementación de sistemas de este tipo, dado que no existe ninguna herramienta disponible que se considere un predictor de riesgo ideal, se valora como alternativa un sistema mixto o mix prospectivo-retrospectivo, a través del cual podría evitarse la persistencia de mecanismos de selección de pacientes debida a las imperfecciones en el nivel de ajuste alcanzado