Search

Abstract Background There is a lack of self-report measures specifically designed for Adjustment disorder. One of the most widely used instruments is the Adjustment Disorder New Module (ADNM), which has already been validated in different countries. The aim of this study was to validate the Spanish version of the ADNM-20 with a non-clinical sample, including a comparison with a high-risk sample. Method 203 participants of the general population (mean age = 33.84) completed an online survey which included the Spanish version of the ADNM, the anxiety and depression measures GAD-7 and PHQ-9, and an assesment of coping strategies using the Brief-COPE. Results The analysis of the scale’s internal structure revealed that a hierarchical model could best fit the data, as well as a bifactorial structure. The scale showed high internal consistency and an adequate differential validity, as differences were observed when comparing the high-risk and non-clinical groups. Positive correlations were also identified between ADNM and depression and anxiety, as well as different correlations with coping strategies. Conclusion This study presents the first validation of the ADNM-20 in Spanish showing good reliability and validity results. In addition, a comparison with a high-risk sample and norms are offered to help future evaluators.

The aim of the research is to examine the biophysical, spatial, functional, and structural components of the pre-Hispanic terracing systems located in the ancient city of Pisaq, considering the impacts of tourism, geological instability, and cultural loss on the ecological and economic value of the terracing system. The methodology includes site analysis, climatology, and an examination of local flora and fauna, supported by digital tools such as QGIS 3.34, Google Earth Pro 2024, and Sun-Path. The results were primarily supported by the use of software tools such as QGIS, AutoCAD 2023, SketchUp 2022, 3D Sun-Path, D5 Render 2024, and Photoshop 2021. The findings include a biophysical analysis related to ecological and economic zoning (EEZ), which determines variables for preservation and reforestation; a spatial analysis measuring the cultivation terraces, with areas ranging from 4.89 ha to 110.20 ha; a functional analysis examining geophysical aspects such as seismic resistance and microclimate effects due to the greenhouse effect; and a constructive analysis that characterizes terrace typologies from an architectural perspective. In conclusion, this analysis evaluates the terracing system of the archaeological park to ensure its preservation and effective management. It also highlights that Inca culture has left a legacy of sustainable architecture, which aligns with the current Sustainable Development Goals (SDGs) (6, 11, 13, 15).

Background: Nursing, as an essential component of contemporary healthcare systems, has undergone significant changes in recent decades, resulting in an increase in research across various fields, such as mental health, well-being, and the development of the discipline itself. Currently, gender stereotypes persist, impacting the social image of the profession and influencing the professional identity of nurses and consequently, their work performance and the quality of care provided. Both public and internal perceptions of the profession are critical factors in these dynamics. Objective: to adapt and validate the Nursing Attitude Questionnaire in a Spanish sample, as it has only been validated in two languages to date. Methods: This study presents descriptive and validity analyses of several measures utilized, such as Nursing Image (NIS), Medical Empathy (JSE-HP), Professional Values (NPVS) and Communication Skills (HP-CSS). Results: Confirmatory factor model analyses indicated that a 21-item hierarchical model provided a superior fit (χ2(df = 1.65, NFI = 0.954, SRMR = 0.065), suggesting the presence of a general factor accounting for five first-order factors. Internal consistency was deemed adequate for the total measures (α = 0.789), though variable for the subscales. Correlations among measures provided evidence of convergent and discriminant validity, highlighting consistent correlation patterns between Attitude toward Nursing and other variables. In addition, significant differences were observed in the Professionalism subscale according to sex, albeit with a medium-low effect size. Scales are presented to facilitate future interpretation of the results in similar samples. Conclusions: The scale validated in this study exhibited overall good psychometric performance, although certain limitations were identified within the subscales. The established scales represent a novel contribution, providing a valuable tool for the comparison of similar samples.

This review explores the state‐of‐the‐art of thermoelectric materials, covering different crystalline structures and material families (e.g., chalcogenides, Zintl phases, skutterudites, clathrates, oxides, half‐Heusler, organic–inorganic composites, metal–organic frameworks, and silicides). It examines their corresponding thermoelectric properties while considering the synthesis methods employed, paying significant attention to those that particularly follow sustainable routes. Additionally, the work addresses current challenges in the field, such as enhancing stability at high temperatures and reducing manufacturing costs. The understanding gained in this field opens avenues for designing more efficient and sustainable devices to convert waste heat into electrical energy, thereby advancing cleaner technologies.

Background: Amyotrophic lateral sclerosis (ALS) is characterized by progressive motor neuron degeneration, with oxidative stress playing a key role. Paraoxonase 1 (PON1) is an antioxidant enzyme that may influence ALS progression. This study aimed to establish a predictive model for the influence of PON1 activity on functionality in ALS patients and explore its relationship with nutrition. Methods: In this observational cross-sectional study, 70 ALS patients underwent assessments of PON1 activity, lipid profile, functional capacity, respiratory function, and heart rate variability. A structural equation model was developed to determine the relationships between variables. Nutritional intake was analyzed in 65 patients. Results: The predictive model showed that PON1 activity and LDL levels positively influenced functionality, both directly and indirectly through respiratory capacity. Heart rate variability moderately predicted functionality independently. HDL levels were not significantly associated with functionality. Weak to moderate correlations were found between PON1 activity and intake of certain nutrients, with positive associations for monounsaturated fats and vitamin D, and negative associations for carbohydrates, proteins, and some micronutrients. Conclusions: PON1 activity appears to play an important role in ALS patient functionality, both directly and through effects on respiratory capacity. However, its relationship with nutritional intake was not strongly evident in this sample population.

El conocimiento que el alumnado actual posee sobre las menciones de los grados de Maestro/a es un campo de interés para saber en qué punto se encuentran con respecto a sus elecciones en relación con sus salidas profesionales. Esta investigación pretende explorar las diferencias de la percepción sobre las menciones en función del sexo, el grado estudiado (Doble grado o Infantil/Primaria) y el curso (2º, 3º, 4º o 5º) y si el hecho de poseer estudios previos musicales influye en la percepción de las menciones. También analizar el papel que tiene en esta percepción el hecho de haber trabajado aspectos musicales en el instituto. Se aplicó un cuestionario a 337 alumnos de los cursos 2º, 3º, 4º y 5º del Grado de Maestro en Educación Infantil, Maestro en Educación Primaria y Doble grado en Maestro en Educación Infantil y Maestro en Educación Primaria de la Universidad Autónoma de Madrid, Universidad de Castilla-La Mancha, Universidad de Salamanca y Universidad de Murcia. Se analizaron los datos en primer lugar realizando los estadísticos previos de las principales medidas del cuestionario. En segundo lugar, se llevó a cabo un Análisis Factorial Exploratorio y por último se estudiaron las diferencias mediante pruebas t de Student. Finalmente, se estudiaron asociaciones entre las medidas de percepción de las menciones y el resto de las medidas categóricas usando pruebas χ2 de Pearson, además, del cálculo de la V de Kramer como índice de asociación. Se concluye insistiendo en la necesidad de más información con respecto a la definición de las menciones y a sus salidas profesionalizantes en el alumnado actual.

Background: The clinical burden of influenza is increasing worldwide. Aging, immunosuppression, and underlying respiratory illness are determinants of poor clinical outcomes, including greater mortality. Bacterial infections seem to be the main reason. Updated information on the role of bacterial infection as the cause of complications would be of value in improving the prognosis of patients with influenza. Methods: A systematic review and meta-analysis were performed by using the PubMed repository using keywords like: Influenza, H1N1, Streptococcus pneumoniae, bacterial coinfection, secondary coinfection, bacterial complications in pneumonia, and seasonal influenza. Only articles written in English were included in publications from 2010 to 2020. The analyses were conducted following the preferred reporting items for systematic review and meta-analyses guidelines. The results were independently validated using a TrinetX database cohort of roughly 4 million patients. Results: We included 135 studies that contained data from 48,259 patients hospitalized with influenza of any age. Bacterial infections were diagnosed in 5391 (11.2%). Streptococcus pneumoniae (30.7%) and Staphylococcus aureus (30.4%) were the most frequent microorganisms, followed by Haemophilus influenzae (7.1%) and Pseudomonas aeruginosa (5.9%). The random-effects model of the meta-analysis indicated that bacterial infections posed a 3.4-fold increased risk of death compared with influenza infection alone. Unexpectedly, asthma was protective (odds ratio 0.8). Conclusion: Bacterial infections diagnosed in 11.2% of patients with influenza increase 3.4-fold the mortality risk. S. pneumoniae, S. aureus, H. influenzae, and P. aeruginosa account for nearly 75% of the cases. Earlier diagnosis and use of antibiotics should improve outcomes in this population.

ABSTRACT Influenza A virus (IAV) infection triggers quantitative and qualitative modifications in lung and intestinal microbiota composition, which contain an important reservoir of antibiotic resistance genes. Analysis of genetic changes is a common practice in studies that analyze microbiota modifications. However, there is little evidence of functional changes linked to such microbiota modifications. This study evaluates some cecal microbiota’s functional changes, comparing sublethal IAV with mock-infected mice. Community-wide phenotypic metabolic profile (Biolog EcoPlatesTM) and relative antibiotic resistance changes to clinically relevant antibiotics (cenoantibiogram) have been performed in this context. Results reveal a temporal association between IAV infection and alterations in nutrient substrate profile usage as well as changes in antibiotic resistance of cecal microbiota. Alterations are transient and predominantly occur at early time points post-IAV infection. There is a functional rebalance in nutrient substrate usage and antibiotic resistance under the established culture conditions, accompanied by a decrease in microbial density of the cecal community on days 5 and 7 after the IAV infection. Our data underline that active IAV infections altering microbial populations are associated with changes in nutrient usage preferences and affect community behaviors toward specific antibiotics. These findings could have implications including activation of nutrient-related metabolic stress at the microbiota community level and additional antibiotic resistance selection mechanisms of clinically relevant infections. IMPORTANCE Influenza virus infection affects both lung and intestinal bacterial community composition. Most of the published analyses focus on the characterization of the microbiota composition changes. Here we assess functional alterations of gut microbiota such as nutrient and antibiotic resistance changes during an acute respiratory tract infection. Upon influenza A virus (IAV) infection, cecal microbiota drops accompanied by a decrease in the ability to metabolize some common nutrients under aerobic conditions. At the same time, the cecal community presents an increase in resistance against clinically relevant antibiotics, particularly cephalosporins. Functional characterization of complex communities presents an additional and necessary element of analysis that nowadays is mainly limited to taxonomic description. The consequences of these functional alterations could affect treatment strategies, especially in multimicrobial infections.

Introduction: Amyotrophic lateral sclerosis (ALS) produces alterations in the autonomic nervous system (ANS), which explains the cardiac manifestations observed in patients. The assessment of heart rate variability (HRV) is what best reflects the activity of the ANS on heart rate. The Polar H7 Bluetooth® device proves to be a non-invasive and much faster technology than existing alternatives for this purpose. Objective: The goal of this study is to determine HRV using Polar H7 Bluetooth technology in ALS patients, comparing the obtained measurements with values from healthy individuals. Method: The sample consisted of 124 participants: 68 diagnosed with ALS and 56 healthy individuals. Using Polar H7 Bluetooth technology and the ELITE HRV application, various HRV measurements were determined for all participants, specifically the HRV index, RMSSD, RMSSD LN, SDNN index, PNN50, LF, HF, LF/HF ratio, HR average, and HF peak frequency. Results: Statistically significant differences were observed between ALS patients and healthy individuals in the HRV index, RMSSD, RMSSD LN, SDNN index, PNN50, HF, and LF, where healthy individuals exhibited higher scores. For the HR average, the ALS group showed a higher value. Values were similar when comparing men and women with ALS, with only a higher HF peak frequency observed in women. Conclusion: The Polar H7 Bluetooth® device is effective in determining heart rate variability alterations in ALS, being a promising prognostic tool for the disease.

Determining the eruption frequency-Magnitude (f-M) relationship for data-limited volcanoes is challenging since it requires a comprehensive eruption record of the past eruptive activity. This is the case for Melimoyu, a long-dormant and data-limited volcano in the Southern Volcanic Zone (SVZ) in Chile with only two confirmed Holocene eruptions (VEI 5). To supplement the eruption records, we identified analogue volcanoes for Melimoyu (i.e., volcanoes that behave similarly and are identified through shared characteristics) using a quantitative and objective approach. Firstly, we compiled a global database containing 181 variables describing the eruptive history, tectonic setting, rock composition, and morphology of 1,428 volcanoes. This database was filtered primarily based on data availability into an input dataset comprising 37 numerical variables for 438 subduction zone volcanoes. Then, we applied Agglomerative Nesting, a bottom-up hierarchical clustering algorithm on three datasets derived from the input dataset: 1) raw data, 2) output from a Principal Component Analysis, and 3) weighted data tuned to minimise the dispersion in the absolute probability per VEI. Lastly, we identified the best set of analogues by analysing the dispersion in the absolute probability per VEI and applying a set of criteria deemed important by the local geological service, SERNAGEOMIN, and VB. Our analysis shows that the raw data generate a low dispersion and the highest number of analogues (n = 20). More than half of these analogues are in the SVZ, suggesting that the tectonic setting plays a key role in the clustering analysis. The eruption f-M relationship modelled from the analogue’s eruption data shows that if Melimoyu has an eruption, there is a 49% probability (50th percentile) of it being VEI≥4. Meanwhile, the annual absolute probability of a VEI≤1, VEI 2, VEI 3, VEI 4, and VEI≥5 eruption at Melimoyu is 4.82 × 10−4, 1.2 × 10−3, 1.45 × 10−4, 9.77 × 10−4, and 8.3 × 10−4 (50th percentile), respectively. Our work shows the importance of using numerical variables to capture the variability across volcanoes and combining quantitative approaches with expert knowledge to assess the suitability of potential analogues. Additionally, this approach allows identifying groups of analogues and can be easily applied to other cases using numerical variables from the global database. Future work will use the analogues to populate an event tree and define eruption source parameters for modelling volcanic hazards at Melimoyu.

Multiple Sclerosis (MS) is a neurodegenerative disease characterized by motor and non-motor symptoms, including emotional distress, anxiety, and depression. These emotional symptoms currently have a pharmacological treatment with limited effectiveness; therefore, it is necessary to delve into their relationship with other psychological, functional, or prefrontal alterations. Additionally, exploring non-pharmacological therapeutic alternatives that have shown benefits in addressing emotional distress in MS patients is essential. Aim: To establish a predictive model for the presence of anxiety and depression in MS patients, based on variables such as psychological well-being, functional activity, and prefrontal symptoms. Additionally, this study aimed to propose non-pharmacological therapeutic alternatives based on this model. Materials and Methods: A descriptive, observational, and cross-sectional study was conducted with a sample of 64 diagnosed MS patients who underwent functional and cognitive assessments using the following questionnaires and scales: Functional Activities Questionnaire (FAQ), Acceptance and Action Questionnaire (AAQ-II), Experiences Questionnaire (EQ), Self-Compassion Scale Short Form (SCS-SF), Beck Depression Inventory II (BDI-II), State-Trait Anxiety Inventory (STAI), and Prefrontal Symptoms Inventory (PSI). Results: The model showed an excellent fit to the data and indicated that psychological well-being was the most significant predictor of the criteria (β = −0.83), followed by functional activity (β = −0.18) and prefrontal symptoms (β = 0.15). The latter two are negatively related to psychological well-being (β = −0.16 and β = −0.75, respectively). Conclusions: Low psychological well-being is the variable that most significantly predicts the presence of anxiety and depression in MS patients, followed by functional activity and prefrontal alterations. Interventions based on mindfulness and acceptance are recommended, along with nutritional interventions such as antioxidant-enriched ketogenic diets and moderate group physical exercise.

The association between obstructive sleep apnea (OSA) and metabolic disorders is well-established; however, the underlying mechanisms that elucidate this relationship remain incompletely understood. Since the liver is a major organ in the maintenance of metabolic homeostasis, we hypothesize that liver dysfunction plays a crucial role in the pathogenesis of metabolic dysfunction associated with obstructive sleep apnea (OSA). Herein, we explored the underlying mechanisms of this association within the liver. Experiments were performed in male Wistar rats fed with a control or high fat (HF) diet (60% lipid-rich) for 12 weeks. Half of the groups were exposed to chronic intermittent hypoxia (CIH) (30 hypoxic (5% O2) cycles, 8 h/day) that mimics OSA, in the last 15 days. Insulin sensitivity and glucose tolerance were assessed. Liver samples were collected for evaluation of lipid deposition, insulin signaling, glucose homeostasis, hypoxia, oxidative stress, antioxidant defenses, mitochondrial biogenesis and inflammation. Both the CIH and HF diet induced dysmetabolism, a state not aggravated in animals submitted to HF plus CIH. CIH aggravates hepatic lipid deposition in obese animals. Hypoxia-inducible factors levels were altered by these stimuli. CIH decreased the levels of oxidative phosphorylation complexes in both groups and the levels of SOD-1. The HF diet reduced mitochondrial density and hepatic antioxidant capacity. The CIH and HF diet produced alterations in cysteine-related thiols and pro-inflammatory markers. The results obtained suggest that hepatic mitochondrial dysfunction and oxidative stress, leading to inflammation, may be significant factors contributing to the development of dysmetabolism associated with OSA.

At present, there is a high demand for carbon (C) sequestration alternatives; thus, understanding tree growth and the efficacy of remote sensing techniques to capture forest plantation ecophysiology is crucial. This study evaluated the effect of contrasting stockings of Gmelina arborea on its initial growth and aboveground Carbon stock, and the efficacy of aerial images obtained using drones to capture the crown cover at different stockings. The results indicated that denser stockings showed greater tree heights and stem diameter increments, contrary to traditional measurements. The C storage capacity of Gmelina arborea was promising, with an aboveground estimated C stock of about 13 Mg ha−1 in 9 months, making it a valuable and promising species for CO2 sequestration under the context of climate change. The use of simple Red-Green-Blue (RGB) cameras and drones to detect and estimate crown areas in young plantations was mainly viable within the commercial range of stockings (500–2000 trees ha−1), and can be used as a powerful tool to better understand tree initial growth. The results showed effective discrimination without weeds independently of the stocking level; however, when weeds were present, the effectiveness decreased. This research provides valuable insights into forest management and improves the understanding of the silviculture behavior of a potential native species for reforestation in the tropics.

El inglés se ha consolidado como el idioma universal dominante, y, por tanto, para una adecuada inserción en el mundo de hoy y al conocimiento que este posee, es sustancial para una persona, adquirirlo. En el Perú, el nivel de dominio del idioma inglés de los egresados de la Educación Básica Regular se encuentra en un nivel básico a pesar de los cientos de horas que el estado imparte como curso, en su malla curricular. Por ello, la presente investigación busca encontrar las causas del bajo dominio del idioma inglés en los estudiantes peruanos de la Educación Básica Regular. Para ello, analiza la metodología de la enseñanza del inglés propuesta por el Ministerio de Educación a los docentes de los colegios, así como un breve análisis a la aplicación del enfoque comunicativo para establecer las competencias con las cuales se mide los logros de los estudiantes. Seguidamente, analiza y compara las políticas estatales para la promoción del idioma inglés con estándares internacionales y con el sector privado. El estudio concluye que el estado no ha logrado darle la debida importancia al curso de inglés, ni ha encontrado los modos de promover la especialización.

Purpose To examine the prevalence of depressive and anxiety symptoms and the corresponding risk factors among pregnant women during the confinement due to the COVID-19 outbreak in Spain. Materials and methods Between 15 April and 14 May 2020, a multicentre cross-sectional survey was performed to study depression, anxiety and resilience in a sample of Spanish pregnant women during the lockdown set up by the Government in response to COVID-19 pandemic outbreak. We designed an anonymous online self-administered questionnaire (https://bit.ly/34RRpq1) that included the Spanish validated versions of the Edinburgh Postpartum Depression Scale (EPDS), the State-Trait Anxiety Inventory (STAI) and the Connor-Davidson Resilience 10-items Scale (CD-RISC-10). Results A total of 514 pregnant women completed the survey. 72.8% had been confined < 40 days and 27.2% between 41 and 60 days. 182 (35.4%) participants scored over 10, with 21.3% scoring over 13 (75th Percentile) in depressive symptoms rates. We found high trait and anxiety scores, with 223 (43.4%) and 227 (44.2%) pregnant women scoring over the trait and state mean scores. Neither depression, anxiety or resilience levels showed any significant correlation with the length of confinement. We found low CD-RISC-10 scores. Conclusions We found a high prevalence of depression and anxiety symptoms during the quarantine, although we did not find an increased prevalence of psychological distress according to length of home confinement. Resilience correlated negatively with depression and anxiety.

Abstract Type 2 diabetes (T2D) is associated with cardiovascular and pulmonary disease. How T2D affects pulmonary endothelial function is not well characterized. We investigated the effects of T2D progression on contractility machinery and endothelial function in the pulmonary and systemic circulation and the mechanisms promoting the dysfunction, using pulmonary artery (PA) and aorta. A high-fat (HF, 3 weeks 60% lipid-rich diet) and a high-fat/high-sucrose (HFHSu, combined 60% lipid-rich diet and 35% sucrose during 25 weeks) groups were used as prediabetes and T2D rat models. We found that T2D progression differently affects endothelial function and vascular contractility in the aorta and PA, with the contractile machinery being altered in the PA and aorta in prediabetes and T2D animals; and endothelial function being affected in both models in the aorta but only affected in the PA of T2D animals, meaning that PA is more resistant than aorta to endothelial dysfunction. Additionally, PA and systemic endothelial dysfunction in diabetic rats were associated with alterations in the nitrergic system and inflammatory pathways. PA dysfunction in T2D involves endothelial wall mineralization. The understanding of the mechanisms behind PA dysfunction in T2D can lead to significant advances in both preventative and therapeutic treatments of pulmonary disease-associated diabetes.

Brand placement in videogames consists in integrating various brands that are not intrusive to give a feeling of reality to the context, as such increasing awareness and recognition of the brand in the mind of the consumer. The goal of this study is to ascertain if brand position, familiarity and congruency influence memorization of brands presented in a racing videogame. An experimental design 2 (position: prominent or subtle) × 2 (congruency: yes or no) × 2 (familiar: yes or no) was used in a sample of 117 participants (M = 20.91 years, SD = 1.75 years). Consumers have better recall and recognition of brand placement when it is familiar. Likewise, there is more memorization of placement that is congruent with the videogame’s subject matter. Nevertheless, it appears that position has no influence on memory. Regarding the three-way interaction, it turns out that the best way to remember brands is when they are familiar to the consumers, congruent with the videogame’s genre, and they are positioned in a prominent position. This article is the first to investigate the interaction between the three main variables that affect the memorization of brand placement.

Inorganic materials can provide a set of tools to decontaminate solid, liquid or air containing viral particles. The use of disinfectants can be limited or not practical in scenarios where continuous cleaning is not feasible. Physicochemical differences between viruses raise the need for effective formulations for all kind of viruses. In the present work we describe two types of antimicrobial inorganic materials: i) a novel soda-lime glass (G3), and ii) kaolin containing metals nanoparticles (Ag or CuO), as materials to disable virus infectivity. Strong antiviral properties can be observed in G3 glass, and kaolin-containing nanoparticle materials showing a reduction of viral infectivity close to 99% in the first 10 ​min of contact of vesicular stomatitis virus (VSV). A potent virucidal activity is also present in G3 and kaolin containing Ag or CuO nanoparticles against all kinds of viruses tested, reducing more than 99% the amount of HSV-1, Adenovirus, VSV, Influenza virus and SARS-CoV-2 exposed to them. Virucidal properties could be explained by a direct interaction of materials with viruses as well as inactivation by the presence of virucidal elements in the material lixiviates. Kaolin-based materials guarantee a controlled release of active nanoparticles with antiviral activity. Current coronavirus crisis highlights the need for new strategies to remove viruses from contaminated areas. We propose these low-cost inorganic materials as useful disinfecting antivirals in the actual or future pandemic threats.

Background & Aims: Patients with primary biliary cholangitis (PBC) exhibit reduced AE2/SLC4A2 gene expression in the liver and peripheral blood mononuclear cells (PBMCs). AE2 encodes a Cl–/HCO3– exchanger involved in biliary bicarbonate secretion and intracellular pH regulation. Reduced AE2 expression in PBC may be pathogenic, as Ae2-knockout mice reproduce characteristic PBC features. Herein, we aimed to identify CpG-methylation abnormalities in AE2 promoter regions that might contribute to the reduced gene transcription in PBC livers and PBMCs. Methods: CpG-cytosine methylation rates were interrogated at 1-base pair resolution in upstream and alternate AE2 promoter regions through pyrosequencing of bisulphite-modified genomic DNA from liver specimens and PBMCs. AE2a and alternative AE2b1 and AE2b2 mRNA levels were measured by real-time PCR. Human lymphoblastoid-T2 cells were treated with 5-aza-2´-deoxycytidine for demethylation assays. Results: AE2 promoters were found to be hypermethylated in PBC livers compared to normal and diseased liver specimens. Receiver operating characteristic (ROC) curve analysis showed that minimal CpG-hypermethylation clusters of 3 AE2a-CpG sites and 4 alternate-AE2b2-CpG sites specifically differentiated PBC from normal and diseased controls, with mean methylation rates inversely correlating with respective transcript levels. Additionally, in PBMCs a minimal cluster of 3 hypermethylated AE2a-CpG sites distinguished PBC from controls, and mean methylation rates correlated negatively with AE2a mRNA levels in these immune cells. Alternate AE2b2/AE2b1 promoters in PBMCs were constitutively hypermethylated, in line with absent alternative mRNA expression in diseased and healthy PBMCs. Demethylation assays treating lymphoblastoid-T2 cells with 5-aza-2´-deoxycytidine triggered AE2b2/AE2b1 expression and upregulated AE2a-promoter expression. Conclusions: Disease-specific hypermethylation of AE2 promoter regions and subsequent downregulation of AE2-gene expression in the liver and PBMCs of patients with PBC might be critically involved in the pathogenesis of this complex disease. Lay summary: Primary biliary cholangitis (PBC) is a chronic immune-associated cholestatic liver disease with unclear complex/multifactorial etiopathogenesis affecting mostly middle-aged women. Patients with PBC exhibit reduced expression of the AE2/SLC4A2 gene. Herein, we found that AE2 promoter regions are hypermethylated in the liver and peripheral blood mononuclear cells of patients with PBC. This increased methylation is associated with downregulated AE2-gene expression, which might contribute to the pathogenesis of PBC. Therefore, novel epigenetic targets may improve treatment in patients with PBC who respond poorly to current pharmacological therapies. Keywords: AE2 chloride/bicarbonate exchanger, cholestasis, DNA methylation, epigenetic repression, primary biliary cholangitis/cirrhosis

The recession suffered in the western world since 2007 has left thousands of people unemployed. One of the countries most affected by unemployment is Spain and specially its young population (34.7%). Considering this context, we try to find out the role of psychological resources, well-being, distress, and eustress, among young employed and unemployed graduates. We worked with a sample of 542 young graduates, of whom 48.3% were unemployed, and those employed held job positions related to their academic background. Our results suggest that: 1) it is somewhat likely for young recent graduates to find a job if they score high in optimism; 2) resilience, optimism, autonomy, self-efficacy, environmental mastery, and overall life satisfaction are deteriorated in unemployment youth, while negative affect is increased in this group; and 3) environmental mastery plays a fundamental role among young unemployed graduates, for its loss is related to distress while its gain is related to eustress. The results of this research are relevant with regards to the implementation of training programs that contribute to the improvement of the well-being and life quality of these unemployed individuals, therefore allowing them to be in a better position to find a job.

Resumen: Introducción: La evaluación del Proceso Asistencial Integrado de Diabetes Mellitus 2 (PAI-DM2) mediante el instrumento para la evaluación de modelos de atención ante la cronicidad (IEMAC-Diabetes) permite el diseño de intervenciones para la mejora de la atención. Objetivo: Analizar la calidad de la atención sanitaria prestada a los pacientes con DM2. Diseño: Estudio cuasiexperimental de tipo antes-después con grupo control no aleatorizado. Emplazamiento: Distritos sanitarios de atención primaria de Sevilla. Participantes: Un total de 12 cupos médicos, 5 centros de atención primaria, seleccionados de manera discrecional. Intervención: Los profesionales de medicina y enfermería de los 12 cupos experimentales participaron en un programa formativo, incluida una estancia externa en el Hospital de Día de Diabetes. Mediciones principales: Número de pacientes incluidos, hemoglobina glucosilada (HbA1c), exploración de pies (EP) y fondo de ojo (FO). Resultados: Se analizaron 1.475 pacientes con DM2. La proporción de pacientes incluidos por cupo fue del 8,3%, siendo mujeres el 45,4%. Al inicio del estudio, la proporción de pacientes con HbA1c

The goal of the current work was to identify an air-side-optimized contact material for La0.3Ca0.7Fe0.7Cr0.3O3−δ (LCFCr) electrodes and a Crofer22APU interconnect for use in reversible solid oxide fuel cells (RSOFCs). LaNi0.6Co0.4−xFexO3 (x = 0–0.3) perovskite-type oxides were investigated in this work. The partial substitution of Co by Fe decreased the thermal expansion coefficient values (TEC) closer to the values of the LCFCr and Crofer 22 APU interconnects. The oxides were synthesized using the glycine–nitrate method and were characterized using X-ray thermodiffraction and 4-probe DC electrical conductivity measurements. Based on the materials characterization results from the Fe-doped oxides investigated here, the LaNi0.6Co0.2Fe0.2O3−δ composition was selected as a good candidate for the contact material, as it exhibited an acceptable electrical conductivity value of 395 S·cm−1 at 800 °C in air and a TEC value of 14.98 × 10−6 K−1 (RT-900 °C).

Chronic sustained hypoxia (CSH), as found in individuals living at a high altitude or in patients suffering respiratory disorders, initiates physiological adaptations such as carotid body stimulation to maintain oxygen levels, but has deleterious effects such as pulmonary hypertension (PH). Obstructive sleep apnea (OSA), a respiratory disorder of increasing prevalence, is characterized by a situation of chronic intermittent hypoxia (CIH). OSA is associated with the development of systemic hypertension and cardiovascular pathologies, due to carotid body and sympathetic overactivation. There is growing evidence that CIH can also compromise the pulmonary circulation, causing pulmonary hypertension in OSA patients and animal models. The aim of this work was to compare hemodynamics, vascular contractility, and L-arginine-NO metabolism in two models of PH in rats, associated with CSH and CIH exposure. We demonstrate that whereas CSH and CIH cause several common effects such as an increased hematocrit, weight loss, and an increase in pulmonary artery pressure (PAP), compared to CIH, CSH seems to have more of an effect on the pulmonary circulation, whereas the effects of CIH are apparently more targeted on the systemic circulation. The results suggest that the endothelial dysfunction evident in pulmonary arteries with both hypoxia protocols are not due to an increase in methylated arginines in these arteries, although an increase in plasma SDMA could contribute to the apparent loss of basal NO-dependent vasodilation and, therefore, the increase in PAP that results from CIH.

Several studies demonstrated a link between obstructive sleep apnea (OSA) and the development of insulin resistance. However, the main event triggering insulin resistance in OSA remains to be clarified. Herein, we investigated the effect of mild and severe chronic intermittent hypoxia (CIH) on whole-body metabolic deregulation and visceral adipose tissue dysfunction. Moreover, we studied the contribution of obesity to CIH-induced dysmetabolic states. Experiments were performed in male Wistar rats submitted to a control and high-fat (HF) diet. Two CIH protocols were tested: A mild CIH paradigm (5/6 hypoxic (5% O2) cycles/h, 10.5 h/day) during 35 days and a severe CIH paradigm (30 hypoxic (5% O2) cycles, 8 h/day) during 15 days. Fasting glycemia, insulinemia, insulin sensitivity, weight, and fat mass were assessed. Adipose tissue hypoxia, inflammation, angiogenesis, oxidative stress, and metabolism were investigated. Mild and severe CIH increased insulin levels and induced whole-body insulin resistance in control animals, effects not associated with weight gain. In control animals, CIH did not modify adipocytes perimeter as well as adipose tissue hypoxia, angiogenesis, inflammation or oxidative stress. In HF animals, severe CIH attenuated the increase in adipocytes perimeter, adipose tissue hypoxia, angiogenesis, and dysmetabolism. In conclusion, adipose tissue dysfunction is not the main trigger for initial dysmetabolism in CIH. CIH in an early stage might have a protective role against the deleterious effects of HF diet on adipose tissue metabolism.

The objectives of the present study were to assess the antibacterial effectiveness of two sodium hypochlorite (NaOCl) concentrations (2.5% and 5.25%) activated by means of two techniques, passive ultrasonic irrigation (PUI) and XP-endo® Finisher (FKG Dentaire SA, La Chaux-de-Fonds, Switzerland) (XPF) against bacteria growth in intracanal mature biofilm. Our aim was to determine if the effect of heating up NaOCl at body temperature (BT) contributed to an improvement of the efficacy of XPF. Sixty-two single-canal human roots previously instrumented were infected with E. faecalis inoculum at 0.5 McFarland and incubated at 37 °C for two weeks. Twelve specimens were randomly selected as positive control, and the remaining fifty were divided into five experimental groups (n = 10). The first two were irrigated with 2.5 vs. 5.25% NaOCl at room temperature (RT), activated with PUI, and the other three were irrigated with XPF. Of these three, two were irrigated using 2.5 vs. 5.25% NaOCl at RT and one was irrigated with 5.25% NaOCl at BT. Our results showed that NaOCl was effective in biofilm removal for all experimental groups (p > 0.05), especially in the groups irrigated with 5.25% NaOCl at room temperature (RT) activated with PUI and the group treated with 5.25% NaOCl at BT with XPF. These groups were the most successful ones (p < 0.001). NaOCl, activated with XPF, was as effective as PUI in biofilm removal from the apical third of the canal when it was used at higher concentration and heated up. This study indicates that XPF only reached the efficacy of PUI when NaOCl was heated up.

Background: Biofilm removal from the root canal during endodontic treatment is necessary to prevent further complications. Irrigation is essential to success. Several irrigants have been proposed without a proper comparison. The aim of the study is to compare the antibacterial capacity of different activated irrigants using passive ultrasonic activation (PUI) or XP-Endo finisher (XPF). Methods: A total of 100 instrumented teeth were incubated in an Eppendorf tube containing 0.5 McFarland of Enterococcus faecalis and incubated for 2 weeks at 37 °C. Roots were divided into 5 groups (n = 20) according to the irrigant type: ethylenediaminetetraacetic acid (EDTA) (17%), ethydronic acid (HEDP) (9%) mixed with 5.25% sodium hypochlorite (NaOCl), EDTA (17%) mixed with 5.25% NaOCl, PBS, and a control group. Each group was divided into two subgroups (n = 10): PUI and XPF. Results: As compared to the untreated control group, the irrigators included in the study had a significant effect in bacteria reduction. The obtained results show HEDP to be the most effective irrigant, since no bacteria were recovered after treatment of this group, followed by EDTA mixed with NaOCl and, finally, the EDTA-irrigated group. Conclusions: HEDP is the best irrigating agent in combination with XPF or PUI file activation to eliminate bacteria in our experimental model.

Background: Relationship of total serum IgE and allergy has been studied to determine if it could be a useful complement allergy diagnosis. Objective: To determine whether serum total IgE is a useful tool in diagnosis of allergy in pediatric population attending outpatient pediatric allergy Regional General Hospital No. 1, Ciudad Obregón, Sonora, Mexico. Material and method: An epidemiological, observational, retrospective, transversal and analytical study of diagnostic test was done including files of children 3-15 years 11 months old, treated in pediatric allergy. For statistical analysis SPSS V-22 was used and through tetrachoric table, sensitivity, specificity of IgE for allergy diagnosis were determined, taking skin tests as a gold standard for diagnosing allergies; p value between the study variables was determined using Student t test. Results: 248 records were studied; male gender was more frequent (59%). IgE sensitivity was 85% and specificity was 20%, cutoff point found was 148 IU/mL, elevated levels of IgE in skin tests to airborne allergens and presence of respiratory symptoms. Conclusions: IgE determination is a useful initial screening test in patients with suspected allergy.

Glioblastoma (GBM) is the most aggressive and frequent primary brain tumor in adults with a median overall survival of 15 months. Tumor recurrence and poor prognosis are related to cancer stem cells (CSCs), which drive resistance to therapies. A common characteristic in GBM is CDKN2A gene loss, located close to the cluster of type I IFN genes at Ch9p21. Newcastle disease virus (NDV) is an avian paramyxovirus with oncolytic and immunostimulatory properties that has been proposed for the treatment of GBM. We have analyzed the CDKN2A-IFN I gene cluster in 1018 glioma tumors and evaluated the NDV oncolytic effect in six GBM CSCs ex vivo and in a mouse model. Our results indicate that more than 50% of GBM patients have some IFN deletion. Moreover, GBM susceptibility to NDV is dependent on the loss of the type I IFN. Infection of GBM with an NDV-expressing influenza virus NS1 protein can overcome the resistance to oncolysis by NDV of type I-competent cells. These results highlight the potential of using NDV vectors in antitumor therapies.

Clinical and experimental evidence indicates a positive correlation between chronic intermittent hypoxia (CIH), increased carotid body (CB) chemosensitivity, enhanced sympatho-respiratory coupling and arterial hypertension and cardiovascular disease. Several groups have reported that both the afferent and efferent arms of the CB chemo-reflex are enhanced in CIH animal models through the oscillatory CB activation by recurrent hypoxia/reoxygenation episodes. Accordingly, CB ablation or denervation results in the reduction of these effects. To date, no studies have determined the effects of CIH treatment in chemo-reflex sensitization in guinea pig, a rodent with a hypofunctional CB and lacking ventilatory responses to hypoxia. We hypothesized that the lack of CB hypoxia response in guinea pig would suppress chemo-reflex sensitization and thereby would attenuate or eliminate respiratory, sympathetic and cardiovascular effects of CIH treatment. The main purpose of this study was to assess if guinea pig CB undergoes overactivation by CIH and to correlate CIH effects on CB chemoreceptors with cardiovascular and respiratory responses to hypoxia. We measured CB secretory activity, ventilatory parameters, systemic arterial pressure and sympathetic activity, basal and in response to acute hypoxia in two groups of animals: control and 30 days CIH exposed male guinea pigs. Our results indicated that CIH guinea pig CB lacks activity elicited by acute hypoxia measured as catecholamine (CA) secretory response or intracellular calcium transients. Plethysmography data showed that only severe hypoxia (7% O2) and hypercapnia (5% CO2) induced a significant increased ventilatory response in CIH animals, together with higher oxygen consumption. Therefore, CIH exposure blunted hyperventilation to hypoxia and hypercapnia normalized to oxygen consumption. Increase in plasma CA and superior cervical ganglion CA content was found, implying a CIH induced sympathetic hyperactivity. CIH promoted cardiovascular adjustments by increasing heart rate and mean arterial blood pressure without cardiac ventricle hypertrophy. In conclusion, CIH does not sensitize CB chemoreceptor response to hypoxia but promotes cardiovascular adjustments probably not mediated by the CB. Guinea pigs could represent an interesting model to elucidate the mechanisms that underlie the long-term effects of CIH exposure to provide evidence for the role of the CB mediating pathological effects in sleep apnea diseases.

Regulatory T (Treg) cells can weaken antitumor immune responses, and inhibition of their function appears to be a promising therapeutic approach in cancer patients. Mice with targeted deletion of the gene encoding the Cl−/HCO3− anion exchanger AE2 (also termed SLC4A2), a membrane-bound carrier involved in intracellular pH regulation, showed a progressive decrease in the number of Treg cells. We therefore challenged AE2 as a potential target for tumor therapy, and generated linear peptides designed to bind the third extracellular loop of AE2, which is crucial for its exchange activity. Peptide p17AE2 exhibited optimal interaction ability and indeed promoted apoptosis in mouse and human Treg cells, while activating effector T-cell function. Interestingly, this linear peptide also induced apoptosis in different types of human leukemia, lymphoma and multiple myeloma cell lines and primary malignant samples, while it showed only moderate effects on normal B lymphocytes. Finally, a macrocyclic AE2 targeting peptide exhibiting increased stability in vivo was effective in mice xenografted with B-cell lymphoma. These data suggest that targeting the anion exchanger AE2 with specific peptides may represent an effective therapeutic approach in B-cell malignancies.

The synthesis of complex functional inorganic materials, such as oxides, can be successfully performed by using microwave irradiation as the source of heat. To achieve this, different routes and set-ups can be used: microwave-assisted synthesis may proceed in the solid state or in solution, aqueous or not, and the set ups may be as simple and accessible as domestic oven or quite sophisticated laboratory equipment. An obvious advantage of this innovative methodology is the considerable reduction in time—minutes rather than hours or days—and, as a consequence, energy saving. No less important is the fact that the particle growth is inhibited and the broad variety of different microwave or microwave-assisted synthesis techniques opens up opportunities for the preparation of inorganic nanoparticles and nanostructures. In this work, various microwave synthesis techniques have been employed: solid-state microwaves, single-mode microwaves using a TE10p cavity and microwave-assisted hydrothermal synthesis. Relevant examples are presented and discussed.

En esta investigación nos preguntamos si existen diferencias en bienestar y en recursos psicológicos en función del nivel educativo. Empleamos una muestra formada por 2902 individuos y comparamos cinco niveles educativos en las variables indicadas. El análisis se hizo mediante ANOVA y mostró diferencias en todas las variables. Los resultados mostraron que tanto los recursos como el bienestar van en aumento a medida que se incrementa el nivel educativo, pero, que este incremento declina en los sujetos que realizaron estudios universitarios, pero, solo, de grado medio. Futuras investigaciones deberán aclarar esta cuestión.

Este estudo explora diferentes recursos pessoais associados ao bem-estar e funcionamento psicológico positivo de estudantes universitários portugueses, tomando na sua análise as variáveis área de curso e género. Foi aplicada a Escala de Funcionamento Psicológico Positivo (EFPP) a 1070 estudantes de diferentes cursos. Observa-se um efeito da interação do género e área científica em 5 dimensões da EFPP. O género diferencia Autonomia, Autoestima e Otimismo (pontuações mais favoráveis nos homens), enquanto a área científica diferencia Resiliência, Autoestima, Propósito na Vida, Desfrute e Otimismo (pontuações superiores nos estudantes de Saúde). Discutem-se implicações para a intervenção no contexto do Ensino Superior.

Cardiotrophin-1 (CT-1) is a cytokine with antiobesity properties and with a role in lipid metabolism regulation and adipose tissue function. The aim of this study was to analyze the molecular mechanisms involved in the lipolytic actions of CT-1 in adipocytes. Recombinant CT-1 (rCT-1) effects on the main proteins and signaling pathways involved in the regulation of lipolysis were evaluated in 3T3-L1 adipocytes and in mice. rCT-1 treatment stimulated basal glycerol release in a concentration- and time-dependent manner in 3T3-L1 adipocytes. rCT-1 (20 ng/ml for 24 h) raised cAMP levels, and in parallel increased protein kinase (PK)A-mediated phosphorylation of perilipin and hormone sensitive lipase (HSL) at Ser660. siRNA knock-down of HSL or PKA, as well as pretreatment with the PKA inhibitor H89, blunted the CT-1-induced lipolysis, suggesting that the lipolytic action of CT-1 in adipocytes is mainly mediated by activation of HSL through the PKA pathway. In ob/ob mice, acute rCT-1 treatment also promoted PKA-mediated phosphorylation of perilipin and HSL at Ser660 and Ser563, and increased adipose triglyceride lipase (desnutrin) content in adipose tissue. These results showed that the ability of CT-1 to regulate the activity of the main lipases underlies the lipolytic action of this cytokine in vitro and in vivo, and could contribute to CT-1 antiobesity effects.

Carotid body (CB) chemoreceptor cells sense arterial blood PO2, generating a neurosecretory response proportional to the intensity of hypoxia. Hydrogen sulfide (H2S) is a physiological gaseous messenger that is proposed to act as an oxygen sensor in CBs, although this concept remains controversial. In the present study we have used the H2S scavenger and vitamin B12 analog hydroxycobalamin (Cbl) as a new tool to investigate the involvement of endogenous H2S in CB oxygen sensing. We observed that the slow-release sulfide donor GYY4137 elicited catecholamine release from isolated whole carotid bodies, and that Cbl prevented this response. Cbl also abolished the rise in [Ca2+]i evoked by 50 µM NaHS in enzymatically dispersed CB glomus cells. Moreover, Cbl markedly inhibited the catecholamine release and [Ca2+]i rise caused by hypoxia in isolated CBs and dispersed glomus cells, respectively, whereas it did not alter these responses when they were evoked by high [K+]e. The L-type Ca2+ channel blocker nifedipine slightly inhibited the rise in CB chemoreceptor cells [Ca2+]i elicited by sulfide, whilst causing a somewhat larger attenuation of the hypoxia-induced Ca2+ signal. We conclude that Cbl is a useful and specific tool for studying the function of H2S in cells. Based on its effects on the CB chemoreceptor cells we propose that endogenous H2S is an amplifier of the hypoxic transduction cascade which acts mainly by stimulating non-L-type Ca2+ channels.

The bibliographical sources are basic elements of care study. The aim of work is the historical reconstruction of caring for women in the Roman era in the early C. I d focusing on the role of design and delivery and the treatment employed. Using qualitative methodology, historical method, focusing on hermeneutics, we study and analyze the characteristics of the design and the therapy used to alleviate the problems of women in the life cycle of reproduction in Roman times, from a constructivist paradigm. The source used is the Key Pliny’s Natural History Second, structured in 37 books. A book dedicated medical XXVI. There are plenty of dedicated therapeutic pharmacopoeia from animals and plants, providing the remedy to be used to facilitate the design, and each of the problems with menstruation, pregnancy and childbirth in women from Roman times Collecting Roman pharmacopoeia of the early century to C. Use plants and animals and natural elements of therapeutic formula that soothe, heal or cure various health problems of women in particular: conception, pregnancy, childbirth, childbirth and menstruation.

With this work we intend to carry out a study about the most frequent terminology used by nurses during the XVI-XVII century. According to classification and meaning of each health-related word, many of them allow us to come to conclusions about the various crafts of healing and caring and subsequently to obtain a good perspective of nursing and medicine during that period. All these terms and words about techniques, procedures, administration of the different medicines, diseases -some of then unknown in our days-, clarify the role of nurses in those centuries, their limitations and recognition. Among the conclusions, the relevance of the different theories, religion and popular beliefs is remarked upon.

Abuse in TV watching means children dedicating too many hours which should be invested in some other activities as games, and sports. It also means pigging out on junk, including in their diet those industrial products highly caloric and poor in nutrients which lead to nutritional upset, obesity and hyper-cholestherolemia. This is a second part of the Report done on TV Habits in a female population between 10 and 14 years of age in Palencia. An Inquiry was shared by girls and parents. The questions and data requested were basically on preferences, intake of food, products, junk, announced on TV at meals time while their preferred programmes were watched. Subjects participated in an anthropometric measurement, level of cholesterol in blood, glucaemia and blood pressure study. Outcomes show a large number of girls watching TV every day at meal times, demand those food announced on TV and they pig out on junk. Those girls with overweight problems do watch TV longer periods of time. Parents, teachers, paediatricians and nurses must be aware of all those negative effects in order to be able to improve well-fare and health behaviour patterns in children population.

Children in Western industrialised countries dedicate more time to watching television (TV) than to any other extra-school activity. Numerous international and national publications have shown that TV favours a sedentary lifestyle and reduces physical activity and school work. This descriptive, transversal study inquires into the TV-watching habits of the school population between 10 and 14 years of age in the city of Plasencia (Extremadura). The study method was based on filling in questionnaires addressed to 494 selected schoolchildren, their parents and their teachers, which referred to TV-watching habits, the time dedicated to physical exercise, sleep and reading, and academic performance. This article shows the results obtained with the 244 girls that participated in the survey. It was proved that girls dedicate 15.9 hours a week to watching TV, an amount of time well above the current recommendations, to the detriment of physical activity, sleep and study, and also that many parents allow their children to watch TV on their own, with no time limits or content selection. People responsible for children’s education must prepare specific programmes to re-educate children’s TV-watching habits.

An experimental and theoretical study on 1,2-dielaidoyl-sn-glycero-3-phosphoethanolamine (DEPE) membranes containing fatty acids (FAs) was performed by means of X-ray diffraction analysis and molecular dynamics (MD) simulations. The study was aimed at understanding the interactions of several structurally related FAs with biomembranes, which is necessary for further rational lipid drug design in membrane-lipid therapy. The main effect of FAs was to promote the formation of a HII phase, despite a stabilization of the coexisting Lα + HII phases. Derivatives of OA exhibited a specific density profile in the direction perpendicular to the bilayer that reflects differences in the relative localization of the carboxylate group within the polar region of the membrane as well as in the degree of membrane penetration of the FA acyl chain. Hydroxyl and methyl substituents at carbon-2 in the FA acyl chain were identified as effective modulators of the position of carboxylate group in the lipid bilayer. Our data highlight the specific potential of each FA in modulating the membrane structure properties.

IGF-I modulates liver tissue homeostasis. It is produced by hepatocytes and signals within the liver through IGF-I receptor expressed on hepatic stellate cells (HSCs). Liver cirrhosis is characterized by marked IGF-I deficiency. Here we compared the effect of two different gene therapy vectors encoding IGF-I as a potential treatment for cirrhotic patients. Rats with carbon tetrachloride-induced liver cirrhosis were treated with controls or with adeno-associated virus 1 (AAV) or simian virus 40 (SV40) vectors expressing IGF-I (AAVIGF-I or SVIGF-I) and molecular and histological studies were performed at 4 days, 8 weeks and 16 weeks. Increased levels of IGF-I were observed in the liver as soon as 4 days after vector administration. Control cirrhotic rats showed increased hepatic expression of pro-inflammatory and pro-fibrogenic factors including transforming growth factor beta (TGFβ), tumor necrosis factor-alpha (TNFα), connective tissue growth factor (CTGF), and vascular endothelial growth factor (VEGF) together with upregulation of α-smooth muscle actin (αSMA), a marker of HSC activation. In IGF-I-treated rats the levels of all these molecules were similar to those of healthy controls by week 8 post-therapy. Of note, the decline of TGFβ, CTGF, VEGF and αSMA expression was more rapid in AAVIGF-I treated animals reaching statistical significance by day 4 post-therapy. IGF-I-treated rats showed similar improvement of liver function tests in parallel with upregulation of hepatocyte nuclear factor 4α (HNF4α), a factor that promotes hepatocellular differentiation. A significant decrease of liver fibrosis, accompanied by upregulation of the hepatoprotective and anti-fibrogenic hepatocyte growth factor (HGF), occurred in all IGF-I-treated rats but complete reversal of liver cirrhosis took place only in AAVIGF-I group. Therefore, AAVIGF-I reverts liver cirrhosis in rats, a capability which deserves clinical testing.

Development of reporter systems for in vivo examination of IFN-β induction or signaling of type I interferon (IFN-I) pathways is of great interest in order to characterize biological responses to different inducers such as viral infections. Several reporter mice have been developed to monitor the induction of both pathways in response to different agonists. However, alternative strategies that do not require transgenic mice breeding have to date not been reported. In addition, detection of these pathways in vivo in animal species other than mice has not yet been addressed. Herein we describe a simple method based on the use of an adeno-associated viral vector (AAV8-3xIRF-ISRE-Luc) containing an IFN-β induction and signaling-sensitive promoter sequence controlling the expression of the reporter gene luciferase. This vector is valid for monitoring IFN-I responses in vivo elicited by diverse stimuli in different organs. Intravenous administration of the vector in C57BL/6 mice and Syrian hamsters was able to detect activation of the IFN pathway in the liver upon systemic treatment with different pro-inflammatory agents and infection with Newcastle disease virus (NDV). In addition, intranasal instillation of AAV8-3xIRF-ISRE-Luc showed a rapid and transient IFN-I response in the respiratory tract of mice infected with the influenza A/PR8/34 virus lacking the NS1 protein. In comparison, this response was delayed and exacerbated in mice infected with influenza A/PR/8 wild type virus. In conclusion, the AAV8-3xIRF-ISRE-Luc vector offers the possibility of detecting IFN-I activation in response to different stimuli and in different animal models with no need for reporter transgenic animals.

Gene therapy of liver cancer covers a variety of gene transfer strategies aimed to the treatment of patients with primary and secondary liver tumors, including gene directed enzyme/pro-drug therapy, inhibition of oncogenes and restoration of tumor-suppressor genes, immunotherapy, anti-angiogenesis and virotherapy. Some of these strategies have reached early clinical development with diverse little success.

Interferon-α is a potent antiviral agent and a vigorous adjuvant in the induction of T-cell responses but its use is limited by hematologic toxicity. Interferon-α alters hematopoietic stem cell dormancy and impairs myelocytic and erythrocytic/megakaryocytic differentiation from hematopoietic progenitors. However, the effect of chronic interferon-α exposure on hematopoietic precursors has still not been well characterized. Here, we transduced the liver of mice with an adenoassociated vector encoding interferon-α to achieve sustained high serum levels of the cytokine. The bone marrow of these animals showed diminished long-term and short-term hematopoietic stem cells, reduction of multipotent progenitor cells, and marked decrease of B cells, but significant increase in the proportion of CD8+ and CD4+CD8+ T cells. Upon adoptive transfer to RAG−/− mice, bone marrow cells from interferon-α-treated animals generated CD4+ and CD8+ T cells while CD19+, CD11b+ and NK1.1+ lineages failed to develop. These effects are associated with the transcriptional downregulation of transcription factors involved in B-cell differentiation and modulation of key factors for T-cell development. Thus, sustained interferon-α exposure causes hematopoietic stem cells exhaustion and drives common lymphoid progenitors towards T-cell generation.

Lipid intake in diet regulates the membrane lipid composition, which in turn controls activities of membrane proteins. There is evidence that fatty acids (FAs) and triacylglycerols (TGs) can alter the phospholipid (PL) mesomorphism. However, the molecular mechanisms involved are not fully understood. This study focuses on the effect of the unsaturation degree of the C-18 FAs, oleic acid (OA), linoleic acid and linolenic acid, and their TGs, triolein (TO), trilinolein, and trilinolenin, on the structural properties of phosphoethanolamine PLs. By means of X-ray diffraction and 31P-NMR spectroscopy, it is shown that both types of molecules stabilize the HII phase in 1,2-dielaidoyl-sn-glycero-3-phosphoethanolamine (DEPE) liposomes. Several structural factors are considered to explain the correlation between the FA unsaturation degree and the onset temperature of the HII phase.It is proposed that TGs could act as lateral spacers between polar DEPE groups, providing an increase in the effective surface area per lipid molecule that would account for the structural parameters of the HII phase. Fluorescence polarization data indicated a fluidification effect of OA on the lamellar phase. TO increased the viscosity of the hydrophobic core with a high effect on the HII phase.

Transforming growth factor β (TGF-β) is a powerful promoter of cancer progression and a key target for antitumor therapy. As cancer cells exhibit active cholesterol metabolism, high density lipoproteins (HDLs) appear as an attractive delivery system for anticancer TGFβ-inhibitory molecules. We constructed a plasmid encoding a potent TGF-β-blocking peptide (P144) linked to apolipoprotein A-I (ApoA-I) through a flexible linker (pApoLinkerP144). The ApoLinkerP144 sequence was then incorporated into a hepatotropic adeno-associated vector (AAVApoLinkerP144). The aim was to induce hepatocytes to produce HDLs containing a modified ApoA-I capable of blocking TGF-β. We observed that transduction of the murine liver with pApoLinkerP144 led to the appearance of a fraction of circulating HDL containing the fusion protein. These HDLs were able to attenuate TGF-β signaling in the liver and to enhance IL-12 -mediated IFN-γ production. Treatment of liver metastasis of MC38 colorectal cancer with AAVApoLinkerP144 resulted in a significant reduction of tumor growth and enhanced expression of IFN-γ and GM-CSF in cancerous tissue. ApoLinkerP144 also delayed MC38 liver metastasis in Rag2-/-IL2rγ-/- immunodeficient mice. This effect was associated with downregulation of TGF-β target genes essential for metastatic niche conditioning. Finally, in a subset of ret transgenic mice, a model of aggressive spontaneous metastatic melanoma, AAVApoLinkerP144 delayed tumor growth in association with increased CD8+ T cell numbers in regional lymph nodes. In conclusion, modification of HDLs to transport TGF-β-blocking molecules is a novel and promising approach to inhibit the growth of liver metastases by immunological and non-immunological mechanisms.

The aim of this study was to analyse the distribution of KIR haplotypes and the KIR2DL2/3 alleles in chronic HCV-infected patients in order to establish the influence on the response to pegylated interferon plus ribavirin classical treatment. The alleles study of previously associated KIR2DL2/3 showed that KIR2DL2*001 was more frequent in non-SVR (NSVR) (42.2% vs. 27.5%, p

It has been shown that the liver of immunodeficient mice can be efficiently repopulated with human hepatocytes when subjected to chronic hepatocellular damage. Mice with such chimeric livers represent useful reagents for medical and clinical studies. However all previously reported models of humanized livers are difficult to implement as they involve cross-breeding of immunodeficient mice with mice exhibiting genetic alterations causing sustained hepatic injury. In this paper we attempted to create chimeric livers by inducing persistent hepatocellular damage in immunodeficient Rag2(-/-) γc(-/-) mice using an adenovirus encoding herpes virus thymidine kinase (AdTk) and two consecutive doses of ganciclovir (GCV). We found that this treatment resulted in hepatocellular damage persisting for at least 10 weeks and enabled efficient engraftment and proliferation within the liver of either human or allogenic hepatocytes. Interestingly, while the nodules generated from the transplanted mouse hepatocytes were well vascularized, the human hepatocytes experienced progressive depolarization and exhibited reduced numbers of murine endothelial cells inside the nodules. In conclusion, AdTk/GCV-induced liver damage licenses the liver of immunodeficient mice for allogenic and xenogenic hepatocyte repopulation. This approach represents a simple alternative strategy for chimeric liver generation using immunodeficient mice without additional genetic manipulation of the germ line.

Recombinant adenoassociated viral vectors (rAAV) have proven to be excellent candidates for gene therapy clinical applications. Recent results showed that cellular immunity to AAV represents a major challenge facing the clinical use of systemic administration of these vectors. Interestingly, no preclinical animal model has previously fully reproduced the clinical findings. The aim of the present work was to enhance the T cell immune response against AAV capsid in mice by the administration of a rAAV expressing the immunostimulatory cytokine IL-12. Our results indicate that although IL-12 expression enhanced the AAV capsid-specific immune response it failed to eliminate transduced hepatocytes and long-term expression was achieved. We found that AAV-mediated transgene expression is altered by IL-12-induced liver inflammation. However, IL-12 expression has no effect over preexisting AAV-mediated transgene expression. IL-12 down-regulates AAV mediated transgene expression via induction of IFN-γ production by NK and T cells, but without altering the transduction efficiency measured by viral genomes. Our results indicate that liver inflammation affects the formation of transcriptionally active AAV vector genomes through an unknown mechanism that can be avoided by the use of DNA-demethylating or anti-inflammatory agents.

OBJECTIVE: Only a limited proportion of patients needing pharmacological control of portal hypertension are hemodynamic responders to propranolol. Here we analyzed the effects of zolmitriptan on portal pressure and its potential interaction with propranolol. METHODS: ZOLMITRIPTAN, PROPRANOLOL OR BOTH WERE TESTED IN TWO RAT MODELS OF PORTAL HYPERTENSION: common bile duct ligation (CBDL) and CCl4-induced cirrhosis. In these animals we measured different hemodynamic parameters including portal venous pressure, arterial renal flow, portal blood flow and cardiac output. We also studied the changes in superior mesenteric artery perfusion pressure and in arterial wall cAMP levels induced by zolmitriptan, propranolol or both. Moreover, we determined the effect of splanchnic sympathectomy on the response of PVP to zolmitriptan. RESULTS: In both models of portal hypertension zolmitriptan induced a dose-dependent transient descent of portal pressure accompanied by reduction of portal flow with only slight decrease in renal flow. In cirrhotic rats, splanchnic sympathectomy intensified and prolonged zolmitriptan-induced portal pressure descent. Also, propranolol caused more intense and durable portal pressure fall when combined with zolmitriptan. Mesenteric artery perfusion pressure peaked for about 1 min upon zolmitriptan administration but showed no change with propranolol. However propranolol enhanced and prolonged the elevation in mesenteric artery perfusion pressure induced by zolmitriptan. In vitro studies showed that propranolol prevented the inhibitory effects of β2-agonists on zolmitriptan-induced vasoconstriction and the combination of propranolol and zolmitriptan significantly reduced the elevation of cAMP caused by β2-agonists. CONCLUSION: Zolmitriptan reduces portal hypertension and non-selective beta-blockers can improve this effect. Combination therapy deserves consideration for patients with portal hypertension failing to respond to non-selective beta-blockers.