Characterizing regional radon-in-air levels in rocks of the Canary Islands (Spain): new data and results.

In this work, a regional-scale strategy to characterize the radon activity levels in the Canary Islands (Spain) is described. The main objectives of this strategy consisted of (1) studying the likely relationship between radon concentration and lithology of the rock matrix through the lithological data of 247 samples from volcanic rocks of the Canary Islands and (2) implementing a series of monitoring sites in the form of boreholes and wells to study the evolution of radon-in-air activity on a daily to yearly timescale.

KLHL3 regulates paracellular chloride transport in the kidney by ubiquitination of claudin-8.

A rare Mendelian syndrome--pseudohypoaldosteronism type II (PHA-II)--features hypertension, hyperkalemia, and metabolic acidosis. Genetic linkage studies and exome sequencing have identified four genes--with no lysine kinase 1 (wnk1), wnk4, Kelch-like 3 (KLHL3), and Cullin 3 (Cul3)--mutations of which all caused PHA-II phenotypes. The previous hypothesis was that the KLHL3-Cul3 ubiquitin complex acted on the wnk4-wnk1 kinase complex to regulate Na(+)/Cl(-) cotransporter (NCC) mediated salt reabsorption in the distal tubules of the kidney. Here, we report the identification of claudin-8 as a previously unidentified physiologic target for KLHL3 and provide an alternative explanation for the collecting duct's role in PHA-II. Using a tissue-specific KO approach, we have found that deletion of claudin-8 in the collecting duct of mouse kidney caused hypotension, hypokalemia, and metabolic alkalosis, an exact mirror image of PHA-II. Mechanistically, the phenotypes in claudin-8 KO animals were caused by disruption of the claudin-8 interaction with claudin-4, the paracellular chloride channel, and delocalization of claudin-4 from the tight junction. In mouse collecting duct cells, knockdown of KLHL3 profoundly increased the paracellular chloride permeability. Mechanistically, KLHL3 was directly bound to claudin-8, and this binding led to the ubiquitination and degradation of claudin-8. The dominant PHA-II mutation in KLHL3 impaired claudin-8 binding, ubiquitination, and degradation. These findings have attested to the concept that the paracellular pathway is physiologically regulated through the ubiquitination pathway, and its deregulation may lead to diseases of electrolyte and blood pressure imbalances.

The Cap1-claudin-4 regulatory pathway is important for renal chloride reabsorption and blood pressure regulation.

The paracellular pathway through the tight junction provides an important route for transepithelial chloride reabsorption in the kidney, which regulates extracellular salt content and blood pressure. Defects in paracellular chloride reabsorption may in theory cause deregulation of blood pressure. However, there is no evidence to prove this theory or to demonstrate the in vivo role of the paracellular pathway in renal chloride handling. Here, using a tissue-specific KO approach, we have revealed a chloride transport pathway in the kidney that requires the tight junction molecule claudin-4. The collecting duct-specific claudin-4 KO animals developed hypotension, hypochloremia, and metabolic alkalosis due to profound renal wasting of chloride. The claudin-4-mediated chloride conductance can be regulated endogenously by a protease-channel-activating protease 1 (cap1). Mechanistically, cap1 regulates claudin-4 intercellular interaction and membrane stability. A putative cap1 cleavage site has been identified in the second extracellular loop of claudin-4, mutation of which abolished its regulation by cap1. The cap1 effects on paracellular chloride permeation can be extended to other proteases such as trypsin, suggesting a general mechanism may also exist for proteases to regulate the tight junction permeabilities. Together, we have discovered a theory that paracellular chloride permeability is physiologically regulated and essential to renal salt homeostasis and blood pressure control.

Neuronal-Targeted TFEB Accelerates Lysosomal Degradation of APP, Reducing Aß Generation and Amyloid Plaque Pathogenesis.

In AD, an imbalance between Aß production and removal drives elevated brain Aß levels and eventual amyloid plaque deposition. APP undergoes nonamyloidogenic processing via α-cleavage at the plasma membrane, amyloidogenic ß- and γ-cleavage within endosomes to generate Aß, or lysosomal degradation in neurons. Considering multiple reports implicating impaired lysosome function as a driver of increased amyloidogenic processing of APP, we explored the efficacy of targeting transcription factor EB (TFEB), a master regulator of lysosomal pathways, to reduce Aß levels. CMV promoter-driven TFEB, transduced via stereotactic hippocampal injections of adeno-associated virus particles in APP/PS1 mice, localized primarily to neuronal nuclei and upregulated lysosome biogenesis. This resulted in reduction of APP protein, the α and ß C-terminal APP fragments (CTFs), and in the steady-state Aß levels in the brain interstitial fluid. In aged mice, total Aß levels and amyloid plaque load were selectively reduced in the TFEB-transduced hippocampi. TFEB transfection in N2a cells stably expressing APP695, stimulated lysosome biogenesis, reduced steady-state levels of APP and α- and ß-CTFs, and attenuated Aß generation by accelerating flux through the endosome-lysosome pathway. Cycloheximide chase assays revealed a shortening of APP half-life with exogenous TFEB expression, which was prevented by concomitant inhibition of lysosomal acidification. These data indicate that TFEB enhances flux through lysosomal degradative pathways to induce APP degradation and reduce Aß generation. Activation of TFEB in neurons is an effective strategy to attenuate Aß generation and attenuate amyloid plaque deposition in AD. SIGNIFICANCE STATEMENT: A key driver for AD pathogenesis is the net balance between production and clearance of Aß, the major component of amyloid plaques. Here we demonstrate that lysosomal degradation of holo-APP influences Aß production by limiting the availability of APP for amyloidogenic processing. Using viral gene transfer of transcription factor EB (TFEB), a master regulator of lysosome biogenesis in neurons of APP/PS1 mice, steady-state levels of APP were reduced, resulting in decreased interstitial fluid Aß levels and attenuated amyloid deposits. These effects were caused by accelerated lysosomal degradation of endocytosed APP, reflected by reduced APP half-life and steady-state levels in TFEB-expressing cells, with resultant decrease in Aß production and release. Additional studies are needed to explore the therapeutic potential of this approach.

Enhancing astrocytic lysosome biogenesis facilitates Aß clearance and attenuates amyloid plaque pathogenesis.

In sporadic Alzheimer's disease (AD), impaired Aß removal contributes to elevated extracellular Aß levels that drive amyloid plaque pathogenesis. Extracellular proteolysis, export across the blood-brain barrier, and cellular uptake facilitate physiologic Aß clearance. Astrocytes can take up and degrade Aß, but it remains unclear whether this function is insufficient in AD or can be enhanced to accelerate Aß removal. Additionally, age-related dysfunction of lysosomes, the major degradative organelles wherein Aß localizes after uptake, has been implicated in amyloid plaque pathogenesis. We tested the hypothesis that enhancing lysosomal function in astrocytes with transcription factor EB (TFEB), a master regulator of lysosome biogenesis, would promote Aß uptake and catabolism and attenuate plaque pathogenesis. Exogenous TFEB localized to the nucleus with transcriptional induction of lysosomal biogenesis and function in vitro. This resulted in significantly accelerated uptake of exogenously applied Aß42, with increased localization to and degradation within lysosomes in C17.2 cells and primary astrocytes, indicating that TFEB is sufficient to coordinately enhance uptake, trafficking, and degradation of Aß. Stereotactic injection of adeno-associated viral particles carrying TFEB driven by a glial fibrillary acidic protein promoter was used to achieve astrocyte-specific expression in the hippocampus of APP/PS1 transgenic mice. Exogenous TFEB localized to astrocyte nuclei and enhanced lysosome function, resulting in reduced Aß levels and shortened half-life in the brain interstitial fluid and reduced amyloid plaque load in the hippocampus compared with control virus-injected mice. Therefore, activation of TFEB in astrocytes is an effective strategy to restore adequate Aß removal and counter amyloid plaque pathogenesis in AD.

Role of phosphatidylinositol clathrin assembly lymphoid-myeloid leukemia (PICALM) in intracellular amyloid precursor protein (APP) processing and amyloid plaque pathogenesis.

One of the pathological hallmarks of Alzheimer disease is the accumulation of amyloid plaques in the extracellular space in the brain. Amyloid plaques are primarily composed of aggregated amyloid ß peptide (Aß), a proteolytic fragment of the transmembrane amyloid precursor protein (APP). For APP to be proteolytically cleaved into Aß, it must be internalized into the cell and trafficked to endosomes where specific protease complexes can cleave APP. Several recent genome-wide association studies have reported that several single nucleotide polymorphisms (SNPs) in the phosphatidylinositol clathrin assembly lymphoid-myeloid leukemia (PICALM) gene were significantly associated with Alzheimer disease, suggesting a role in APP endocytosis and Aß generation. Here, we show that PICALM co-localizes with APP in intracellular vesicles of N2a-APP cells after endocytosis is initiated. PICALM knockdown resulted in reduced APP internalization and Aß generation. Conversely, PICALM overexpression increased APP internalization and Aß production. In vivo, PICALM was found to be expressed in neurons and co-localized with APP throughout the cortex and hippocampus in APP/PS1 mice. PICALM expression was altered using AAV8 gene transfer of PICALM shRNA or PICALM cDNA into the hippocampus of 6-month-old APP/PS1 mice. PICALM knockdown decreased soluble and insoluble Aß levels and amyloid plaque load in the hippocampus. Conversely, PICALM overexpression increased Aß levels and amyloid plaque load. These data indicate that PICALM, an adaptor protein involved in clathrin-mediated endocytosis, regulates APP internalization and subsequent Aß generation. PICALM contributes to amyloid plaque load in brain likely via its effect on Aß metabolism.

Analysis for Candida albicans in samples of intravenous lipids administered to premature infants with the goal of optimizing the use of the 1.2 µm filter.

Introduction: Introduction: parenteral nutrition is a mixture of macro and micronutrients necessary for the premature infant who cannot be fed enterally. The binary mixture contains carbohydrates, amino acids and micronutrients in one bag and intravenous lipids in another. The latter are more susceptible to microbial contamination, especially by Candida albicans. For this reason, many professional associations typically recommend the use of a single filter in line "Y"; however, this has not yet become standard hospital practice. Aim: to determine the presence of Candida albicans in devices that contain intravenous lipids used in neonates and relate it to the correct use of the 1.2 µm filter. Method: three groups of samples consisting of the remains of a lipid solution (ML) administered to the premature patient for 24 h seeded on Sabouraud agar organized as follows: (ML1), lipid solution obtained directly from the ethinyl vinyl acetate bag were evaluated. (ML2): filtered lipid solution with a 1.2 µm device connected directly to the catheter. (ML3): solution of lipids intentionally contaminated with Candida and subsequently filtered. Results: Candida albicans was not detected in any of the filtered simples (ML2 and ML3) and also not detected in any of the unfiltered simples (ML1). Conclusions: there was no presence of Candida albicans in the lipid solutions used directly with a 1.2 µm filter, however, the use of a single 1.2 µm filter in line "Y" is recommended according to international standards to save the health system.

Introducción: Introducción: la nutrición parenteral es una mezcla de macro y micronutrientes necesarios para el prematuro que no puede alimentarse por vía enteral. La mezcla binaria contiene en una bolsa carbohidratos, proteínas y micronutrientes y en otra los lípidos intravenosos, que son más susceptibles de contaminación microbiana especialmente por Candida albicans, por lo que asociaciones profesionales recomiendan el uso en línea "y" de un solo filtro, lo que no es práctica habitual hospitalaria. Objetivo: determinar la presencia de Candida albicans en dispositivos que contienen lípidos intravenosos usados en neonatos y relacionarla con el uso correcto del filtro de 1.2 µm. Método: se evaluaron tres grupos de muestras constituidas por los restos de una solución de lípidos (ML) administrados al paciente prematuro durante 24 h. sembradas en agar Sabouraud organizadas en: (ML1): solución de lípidos obtenida directamente de la bolsa de etinil vinil acetato (EVA). (ML2): solución de lípidos filtrados con dispositivo de 1.2 µm. conectado directamente al catéter (práctica observada). (ML3): solución de lípidos contaminados con Candida que fueron sembrados en agar Sabouraud y posteriormente filtrados. Resultados: el 100 % de las muestras filtradas (ML2) y (ML3) como se esperaba, no presentaron Candida albicans, el 100 % de las muestras (ML1) que no fueron filtradas, tampoco presentaron Candida albicans. Conclusiones: no hubo presencia de Candida albicans en las soluciones lipídicas utilizadas directamente con filtro 1.2 µm, sin embargo, se recomienda el uso de un solo filtro 1.2 µm en línea "Y" según las normas internacionales para el ahorro del Sistema sanitario.

Chilled raw milk quality: a case study in Zona da Mata region, Minas Gerais State, Brazil / Qualidade do leite cru refrigerado: um estudo de caso na região da Zona da Mata de Minas Gerais

ABSTRACT: This paper aimed to analyze the evolution of the quality of raw milk produced by producers in the Zona da Mata region, in the state of Minas Gerais, between 2012 and 2018. For this purpose, we used the linear mixed-effects model to analyze the monthly evolution of the results of milk composition indicators (fat, protein, and defatted dry extract - DDE), somatic cell count - SCC, and total bacterial count - TBC, from the official monthly registry of 94 milk producers, suppliers of a region's dairy. Results indicate a continuous reduction in the milk composition indicators between 2014 and 2018. For the SCC and TBC indicators, we identified only one-off reductions. The supply of a larger volume of milk was associated with increased TBC. Seasonality influenced all quality indicators analyzed. According to these results, we concluded that the analyzed quality of chilled raw milk offered by producers was proven worse from 2012 to 2018, despite the actions implemented by the National Milk Quality Improvement Program - PNMQL.

RESUMO: Este artigo objetivou analisar a evolução da qualidade do leite cru refrigerado produzido por produtores da região da Zona da Mata, do Estado de Minas Gerais, entre os anos de 2012 e 2018. Para tal, foi utilizado o modelo linear de efeitos mistos, a fim de analisar a evolução mensal dos resultados de indicadores de composição do leite (gordura, proteína e extrato seco desengordurado - ESD), contagem de células somáticas - CCS, e contagem bacteriana total - CBT, provenientes de análises oficiais mensais de 94 produtores de leite, fornecedores de um laticínio da região. Os resultados indicaram uma redução contínua dos indicadores de composição do leite entre os anos de 2014 e 2018. Para os indicadores de CCS e CBT foram observadas apenas reduções pontuais. O fornecimento de um maior volume de leite foi associado ao aumento da CBT. A sazonalidade influenciou todos os indicadores de qualidade analisados. De posse desses resultados, pode-se afirmar que, apesar das ações implementadas pelo Programa Nacional de Melhoria de Qualidade do Leite - PNMQL, a qualidade do leite cru refrigerado, proveniente dos produtores avaliados, piorou ao longo do período em análise.

Hyperglycemia modulates extracellular amyloid-ß concentrations and neuronal activity in vivo.

Epidemiological studies show that patients with type 2 diabetes (T2DM) and individuals with a diabetes-independent elevation in blood glucose have an increased risk for developing dementia, specifically dementia due to Alzheimer's disease (AD). These observations suggest that abnormal glucose metabolism likely plays a role in some aspects of AD pathogenesis, leading us to investigate the link between aberrant glucose metabolism, T2DM, and AD in murine models. Here, we combined two techniques ­ glucose clamps and in vivo microdialysis ­ as a means to dynamically modulate blood glucose levels in awake, freely moving mice while measuring real-time changes in amyloid-ß (Aß), glucose, and lactate within the hippocampal interstitial fluid (ISF). In a murine model of AD, induction of acute hyperglycemia in young animals increased ISF Aß production and ISF lactate, which serves as a marker of neuronal activity. These effects were exacerbated in aged AD mice with marked Aß plaque pathology. Inward rectifying, ATP-sensitive potassium (K(ATP)) channels mediated the response to elevated glucose levels, as pharmacological manipulation of K(ATP) channels in the hippocampus altered both ISF Aß levels and neuronal activity. Taken together, these results suggest that K(ATP) channel activation mediates the response of hippocampal neurons to hyperglycemia by coupling metabolism with neuronal activity and ISF Aß levels.

Caracterização e desempenho logístico das indústrias laticinistas da Zona da Mata e Campo das Vertentes em Minas Gerais, Brasil / Characterization and logistics performance of dairy industries located at Zona da Mata and Campo das Vertentes in Minas Gerais, Brazil

Objetivando avaliar a atividade logística nas empresas do setor laticinista atuantes nas regiões Zona da Mata e Campo das Vertentes no estado de Minas Gerais, foram promovidas pesquisas com 40 indústrias que representam o setor avaliado. Os questionários respondidos pela gerência das empresas continha variáveis que possibilitaram caracterizar as empresas quanto à sua produção, logística de captação e distribuição, gestão interna, além de mercado e comercialização. O uso da análise estatística multivariada fatorial foi de grande relevância ao possibilitar a construção de um índice que representou o desempenho logístico de cada uma das empresas, sendo também utilizada para realizar o agrupamento dessas indústrias em 4 grupos que continham características similares. O índice logístico (IL) mostrou que as empresas estudadas da região precisam melhorar as atividades que constituíram sua medida, a fim de tornar os aspectos logísticos um meio de vantagem competitiva. Além disso, os grupos formados também possibilitaram avaliar que não é suficiente atuar na melhoria de apenas uma parte das atividades logísticas, mas sim no conjunto de variáveis que pode influenciar positivamente no resultado final no quesito competitividade e liderança logística.

In order to evaluate the logistics activity in the dairy companies operating in Zona da Mata and Campo das Vertentes in Minas Gerais, Brazil a research has been promoted on 40 industries that represent the sector assessed. The questionnaire answered by company employees, contained variables that allowed obtaining information about company identification, gathering and distribution logistics, internal management, marketing and commercialization. Factorial statistical analysis was very important to enable the construction of an index representing the logistics performance of each company and was also used to perform the grouping of these industries into 4 groups containing similar characteristics. The index logistics (IL) showed that companies in the study area still need to improve their activities which were measured to make the logistics a competitive advantage. Moreover, the groups formed also allowed to evaluate that is not enough the improvement at only a part of logistics activities, but is necessary to improve the set of variables that can positively influence the final result in the logistics competitiveness and leadership.

O processo de reconhecimento das indicações geográficas de alimentos e bebidas brasileiras: regulamento de uso, delimitação da área e diferenciação do produto / The process of geographical indications recognition for food and beverages in Brazil: regulation of use, delimitation aerea and product differentiation

As denominações de origem e as indicações de procedência compõem as espécies de indicação geográfica (IG) previstas pela legislação brasileira e têm sido vistas como formas de mobilização e valorização dos territórios e de agregação de valor aos produtos típicos e de qualidade diferenciada. O presente trabalho teve como objetivo caracterizar a condução das etapas de elaboração do regulamento de uso, delimitação da área e comprovação da notoriedade e da relação entre produto e meio geográfico das IGs brasileiras. A metodologia adotada foi pesquisa survey, realizada por meio de questionários semiestruturados. Os resultados revelaram que, apesar das lacunas da legislação brasileira sobre IGs, vários órgãos têm viabilizado o processo de construção das IGs brasileiras atuando de forma decisiva nas etapas de elaboração do regulamento de uso (universidades, ONGs, INPI), delimitação da área (universidades, Embrapa, Emater, IMA), comprovação da notoriedade (SEBRAE) e da relação entre meio geográfico e qualidade (universidades). A comprovação da influência do território nas características qualitativas do produto constitui, atualmente, o principal entrave ao desenvolvimento das denominações de origem no País.

The appellations of origin and indication of source are species of geographical indication (GI) provided by the Brazilian industrial property law that could be seen as one of the ways of mobilization and recovering of the rural territories adding value to local products with differentiated quality. This study aimed to characterize the stages of the construction of the regulations of use of GI, the geographical area delimitation, the evidence of reputation and the relationship between product and geographical environment of GIs in Brazil. The survey research was the methodology adopted conducted through semi-structured questionnaires. The results showed that despite the shortcomings of the Brazilian legislation on GIs, several organs has enabled the construction process of the Brazilian GIs acting decisively on the elaboration steps of the regulation of use (universities, NGOs, INPI), on the area delimitation (universities , Embrapa, Emater, IMA), on the evidence of reputation (SEBRAE) and on the relationship between geographical environment and quality (universities). The proof of the influence of the geographical environment on the product qualitative characteristics is currently the main obstacle to the development of appellations of origin in the country.

Indicação geográfica de alimentos e bebidas no Brasil e na União Europeia / Geographical indication of food and beverages in Brazil and European Union

Entre os direitos relativos à propriedade intelectual, a indicação geográfica (IG) surge como um meio de fomentar o desenvolvimento socioeconômico de uma sociedade. Alguns países, especialmente os europeus, há tempos vêm utilizando a proteção jurídica proporcionada pelo registro das IGs como forma de tornar seus produtos mais competitivos e desenvolver regiões menos favorecidas. No Brasil, o tema é, ainda, recente e necessita ser melhor compreendido, razão pela qual é abordado nesta revisão.

Among rights related to intellectual property, the geographical indication (GI) comes up as mean to stimulate the socioeconomic development of a society. Some countries, especially in Europe, for long have been using legal protection provided by GI certification as a way to make their products more competitive, contributing to the development of less favored regions. In Brazil, the subject is, still, recent and needs to be studied in order to be better understood. This justifies a comprehensive review.