Surface Bragg gratings of proteins patterned on integrated waveguides for (bio)chemical analysis.

The incorporation of biomacromolecules onto silicon waveguiding microstructures constitutes a growing trend that pushes towards compact and miniaturized biosensing systems. This paper presents the integration of one-dimensional periodic nanostructures of proteins on the surface of micrometric silicon waveguides for transducing binding events between biomacromolecules. The study demonstrates this new bioanalytical principle by experimental results and theoretical calculations, and proves that rib waveguides (1--1.6-µm width) together with protein gratings (495--515-nm period) display suitable spectral responses for this optical biosensing system. Protein assemblies of bovine serum albumin are fabricated on the surface of silicon nitride waveguides, characterized by electron microscopy, and their response is measured by optical frequency domain reflectometry along the fabrication process and the subsequent stages of the biorecognition assays. Detection and quantification limits of 0.3 and 3.7 µg·mL-1, respectively, of specific antibodies are inferred from experimental dose-response curves. Among other interesting features, the results of this study point towards new miniaturized and integrated sensors for label-free bioanalysis.

Vitamin E protects from lipid peroxidation during winter stress in the seagrass Cymodocea nodosa.

MAIN CONCLUSION: Adjustments in the antenna size and α-tocopherol contents provide protection from sustained damage in leaves of a seagrass, while low vitamin E contents appear to be enough to protect rhizomes (which appear to be more cold tolerant than leaves). Despite low temperatures can adversely affect the proper growth and development of marine angiosperms, by, among other processes, increasing reactive oxygen species production and causing oxidative damage to lipid membranes, the role of vitamin E in seagrasses, such as Cymodocea nodosa has not been explored thus far. Here, we aimed to better understand the possible role of this chain-breaking (peroxyl radical-trapping) antioxidant in response to low temperatures, and most particularly in relation to the occurrence of photo-inhibition and lipid peroxidation. Low temperatures caused an important desiccation of leaves, but not of rhizomes, which were much more tolerant to cold stress than leaves. Cold stress during winter was associated with chlorophyll loss and transient photo-inhibition, as indicated by reversible reductions in the Fv/Fm ratio. Adjustments in pigment antenna size and vitamin E contents per unit of chlorophyll during winter may help protect the photosynthetic apparatus from sustained photo-inhibitory damage and lipid peroxidation events in leaves. Rhizomes also accumulated significant amounts of vitamin E, although to a much lesser extent than leaves, and kept protected from lipid peroxidation during winter, as indicated by malondialdehyde contents, a product from secondary lipid peroxidation. It is concluded that vitamin E can help protect both leaves and rhizomes from lipid peroxidation, although cold stress during winter can cause transient photo-inhibition of the photosynthetic apparatus, in C. nodosa.

Cytokine production in thyroid eye disease: in vitro effects of dexamethasone and IL-6 blockade with tocilizumab.

PURPOSE: To compare IL-6, sIL-6R, and IL-17 secretion in peripheral blood mononuclear cells (PBMCs) cultured with tocilizumab (a humanized monoclonal antibody against the interleukin-6 receptor), dexamethasone, and placebo, obtained from patients with thyroid eye disease (TED) and healthy controls. METHODS: The study was a prospective proof of concept test. We cultured peripheral blood mononuclear cells from TED patients and healthy controls with tocilizumab, dexamethasone, and placebo. IL-6, sIL-6R, and IL-17 levels in supernatants obtained from PBMCs cultures were analyzed by ELISA. RESULTS: We included seventeen patients with thyroid eye disease (12 females and five males). The mean age was 49 years. Both dexamethasone and tocilizumab influenced IL-6 and IL-6Rs levels in patients' group. Supernatants obtained from PBMCs treated with dexamethasone showed 77.2% and 82.8% lower IL-6 levels compared with those cultured with placebo and tocilizumab, respectively. Furthermore, overnight culture of PBMCs with dexamethasone showed significantly lower sIL-6R secretion compared with untreated (33.71%, p = 0.04) and tocilizumab treated (58.21%, p = 0.01) PBMCs. Neither dexamethasone nor tocilizumab affected IL-17 concentrations in PBMCs cultures. CONCLUSIONS: Both dexamethasone and tocilizumab affect the IL-6/sIL-6R system. Specifically, dexamethasone reduces and tocilizumab increases the levels of these cytokines in PBMCs cultures. These results strengthen the molecular rationale for interrogating the efficacy of tocilizumab in steroid-resistant TED, as IL-6 seems to be a common target for both anti-IL-6R antibody and steroids.

JAK2 mediates lung fibrosis, pulmonary vascular remodelling and hypertension in idiopathic pulmonary fibrosis: an experimental study.

BACKGROUND: Pulmonary hypertension (PH) is a common disorder in patients with idiopathic pulmonary fibrosis (IPF) and portends a poor prognosis. Recent studies using vasodilators approved for PH have failed in improving IPF mainly due to ventilation (V)/perfusion (Q) mismatching and oxygen desaturation. Janus kinase type 2 (JAK2) is a non-receptor tyrosine kinase activated by a broad spectrum of profibrotic and vasoactive mediators, but its role in PH associated to PH is unknown. OBJECTIVE: The study of JAK2 as potential target to treat PH in IPF. METHODS AND RESULTS: JAK2 expression was increased in pulmonary arteries (PAs) from IPF (n=10; 1.93-fold; P=0.0011) and IPF+PH (n=9; 2.65-fold; P<0.0001) compared with PA from control subjects (n=10). PA remodelling was evaluated in human pulmonary artery endothelial cells (HPAECs) and human pulmonary artery smooth muscle cells (HPASMCs) from patients with IPF in vitro treated with the JAK2 inhibitor JSI-124 or siRNA-JAK2 and stimulated with transforming growth factor beta. Both JSI-124 and siRNA-JAK2 inhibited the HPAEC to mesenchymal transition and the HPASMCs to myofibroblast transition and proliferation. JAK2 inhibition induced small PA relaxation in precision-cut lung slice experiments. PA relaxation was dependent of the large conductance calcium-activated potassium channel (BKCa). JAK2 inhibition activated BKCa channels and reduced intracellular Ca2+. JSI-124 1 mg/kg/day, reduced bleomycin-induced lung fibrosis, PA remodelling, right ventricular hypertrophy, PA hypertension and V/Q mismatching in rats. The animal studies followed the ARRIVE guidelines. CONCLUSIONS: JAK2 participates in PA remodelling and tension and may be an attractive target to treat IPF associated to PH.

A Composite of Functional Status and Pneumonia Severity Index Improves the Prediction of Pneumonia Mortality in Older Patients.

BACKGROUND: The baseline health status may be a determinant of interest in the evolution of pneumonia. OBJECTIVE: Our objective was to assess the predictive ability of community-acquired pneumonia (CAP) mortality by combining the Barthel Index (BI) and Pneumonia Severity Index (PSI) in patients aged ≥ 65 years. DESIGN, PATIENTS AND MAIN MEASURES: In this prospective, observational, multicenter analysis of comorbidities, the clinical data, additional examinations and severity of CAP were measured by the PSI and functional status by the BI. Two multivariable models were generated: Model 1 including the PSI and BI and model 2 with PSI plus BI stratified categorically. KEY RESULTS: The total population was 1919 patients, of whom 61% had severe pneumonia (PSI IV-V) and 40.4% had some degree of dependence (BI ≤ 90 points). Mortality in the PSI V-IV group was 12.5%. Some degree of dependence was associated with increased mortality in both the mild (7.2% vs. 3.2%; p = 0.016) and severe (14% vs. 3.3%; p < 0.001) pneumonia groups. The combination of PSI IV-V and BI ≤ 90 was the greatest risk factor for mortality (aOR 4.17; 95% CI 2.48 to 7.02) in our series. CONCLUSIONS: The use of a bimodal model to assess CAP mortality (PSI + BI) provides more accurate prognostic information than the use of each index separately.

Patterned Biolayers of Protein Antigens for Label-Free Biosensing in Cow Milk Allergy.

This paper focuses on creating one-dimensional diffractive grooved structures of antigen proteins on glass substrates for the label-free detection of antibodies to dairy allergens. In particular, the fabrication of protein structures is carried out by combining microcontact printing with physisorption, imines coupling, and thiol-ene click chemistry. The work first sets up these patterning methods and discusses and compares the main aspects involved in them (structure, biolayer thickness, functionality, stability). Homogeneous periodic submicron structures of proteins are created and characterized by diffractive measurements, AFM, FESEM, and fluorescence scanning. Then, this patterning method is applied to proteins involved in cow milk allergy, and the resulting structures are implemented as optical transducers to sense specific immunoglobulins G. In particular, gratings of bovine serum albumin, casein, and ß-lactoglobulin are created and assessed, reaching limits of detection in the range of 30-45 ng·mL-1 of unlabeled antibodies by diffractive biosensing.

Surface Micro-Patterned Biofunctionalized Hydrogel for Direct Nucleic Acid Hybridization Detection.

The present research is focused on the development of a biofunctionalized hydrogel with a surface diffractive micropattern as a label-free biosensing platform. The biosensors described in this paper were fabricated with a holographic recording of polyethylene terephthalate (PET) surface micro-structures, which were then transferred into a hydrogel material. Acrylamide-based hydrogels were obtained with free radical polymerization, and propargyl acrylate was added as a comonomer, which allowed for covalent immobilization of thiolated oligonucleotide probes into the hydrogel network, via thiol-yne photoclick chemistry. The comonomer was shown to significantly contribute to the immobilization of the probes based on fluorescence imaging. Two different immobilization approaches were demonstrated: during or after hydrogel synthesis. The second approach showed better loading capacity of the bioreceptor groups. Diffraction efficiency measurements of hydrogel gratings at 532 nm showed a selective response reaching a limit of detection in the complementary DNA strand of 2.47 µM. The label-free biosensor as designed could significantly contribute to direct and accurate analysis in medical diagnosis as it is cheap, easy to fabricate, and works without the need for further reagents.

Parental Involvement in Homework During Covid-19 Confinement.

BACKGROUND: The purpose of this study was to compare parents' involvement in students' homework during COVID-19 confinement and their involvement in normal circumstances. METHOD: 628 parents of elementary and high school students completed a questionnaire about their family and family involvement in their children's homework. To address the objective, data were examined using mean differences and analysis of variance. RESULTS: As predicted, the COVID-19 situation led to more time spent doing homework for students, as well as higher levels of conflict and stress within the family. In addition, parents reported having increased their homework management during confinement in terms of emotional and motivational support for their children compared to normal circumstances. Finally, the data showed more conflicts and family involvement in the families of children with special needs compared to families of children without special needs. CONCLUSIONS: The time spent in confinement posed many challenges to families' routines and the data indicated a lack of resources available for families to manage their children's tasks, as well as an increase in family stress and conflicts.

Validation of the Shortened Version of the Metacognitive Awareness Inventory in Spanish Uiversity Students.

BACKGROUND: The Metacognitive Awareness Inventory (MAI) is used all over the world to identify metacognitive components that are relevant to learning. However, there is not enough evidence confirming its factorial structure or the suitability of its Shortened Version, proposed by various authors. Also, to date, the MAI has not been validated in the Spanish context. METHOD: After adapting the MAI to Spanish, it was administered to 1076 university students from different regions of Spain. Different structures of the MAI Shortened Version, with 19 items, were compared with the structures of the original 52-item MAI. Indexes of validity and reliability were analyzed, considering the factorial structure and relationships to other validated questionnaires. RESULTS: Only the MAI Shortened Version that differentiates between knowledge and regulation of cognition achieved a good fit. These two scales showed good convergent and divergent validity, high criterion validity in relation to academic achievement, high test- retest reliability, and high internal reliability. CONCLUSIONS: results support the traditional differentiation between knowledge of cognition and regulation of cognition, but only for the Shortened Version of the MAI. This instrument allows quick evaluations and identification of these components in Spanish contexts with adequate metric properties.

Denaturing for Nanoarchitectonics: Local and Periodic UV-Laser Photodeactivation of Protein Biolayers to Create Functional Patterns for Biosensing.

The nanostructuration of biolayers has become a paradigm for exploiting nanoscopic light-matter phenomena for biosensing, among other biomedical purposes. In this work, we present a photopatterning method to create periodic structures of biomacromolecules based on a local and periodic mild denaturation of protein biolayers mediated by UV-laser irradiation. These nanostructures are constituted by a periodic modulation of the protein activity, so they are free of topographic and compositional changes along the pattern. Herein, we introduce the approach, explore the patterning parameters, characterize the resulting structures, and assess their overall homogeneity. This UV-based patterning principle has proven to be an easy, cost-effective, and fast way to fabricate large areas of homogeneous one-dimensional protein patterns (2 min, 15 × 1.2 mm, relative standard deviation ≃ 16%). This work also investigates the implementation of these protein patterns as transducers for diffractive biosensing. Using a model immunoassay, these patterns have demonstrated negligible signal contributions from non-specific bindings and comparable experimental limits of detection in buffer media and in human serum (53 and 36 ng·mL-1 of unlabeled IgG, respectively).

Hypoxemia adds to the CURB-65 pneumonia severity score in hospitalized patients with mild pneumonia.

BACKGROUND: Hypoxemia may influence the prognosis of patients with mild pneumonia, regardless of the initial CURB-65 score (confusion, blood urea nitrogen > 20 mg/dL, respiratory rate > 30 breaths/min, blood pressure < 90/60 mm Hg, and age ≥ 65 y). OBJECTIVE: To determine the risk factors associated with hypoxemia and the influence of hypoxemia on clinical outcomes in hospitalized patients with mild pneumonia. METHODS: We performed a multicenter prospective cohort study of 585 consecutive hospitalized patients with mild pneumonia (CURB-65 groups 0 and 1). We stratified the patients according to the presence of hypoxemia, defined as a P(aO(2))/F(IO(2)) < 300 mm Hg on admission. We assessed the risk factors associated with hypoxemia, hypoxemia's influence on the course of pneumonia, and clinical outcomes (mortality, hospital stay, and need for intensive care unit admission), with multivariable regression. RESULTS: Fifty percent of the patients (294 cases) had hypoxemia on admission. The risk factors independently associated with hypoxemia were: bilateral radiological involvement (odds ratio 2.8, 95% CI 1.1-7.5), history of COPD (odds ratio 2.5, 95% CI 1.4-4.3), and hypoalbuminemia (odds ratio 2.0, 95% CI 1.1-3.5). The hypoxemic patients had longer hospital stay, higher intensive care unit admission rate, higher rate of severe sepsis, and higher mortality than the non-hypoxemic patients. CONCLUSIONS: Hypoxemia in patients with mild pneumonia is independently associated with several adverse clinical and radiological variables, and the hypoxemic patients had worse clinical outcomes than the non-hypoxemic patients. Therefore, additional attention should be paid to the presence of hypoxemia, regardless of a low CURB-65 score.

Two cases of drug-induced orbital inflammatory disease.

Orbital inflammatory disease (OID) includes all inflammatory processes affecting the orbit. Although several aetiologies are recognised, a cause may not be elucidated. We describe 2 cases in which drugs (hyaluronidase and zoledronic acid) were the cause of OID. In patients with a clinical picture of OID simulating an orbital cellulitis, the recent drug history should be considered as a possible aetiology, and treatment with steroids with or without a biopsy should be considered after an infection has been excluded.

What Predicts Life Satisfaction in Children and Adolescents with Attention-Deficit/Hyperactivity Disorder (ADHD)? A Study from Parent and Child/Adolescent Perspectives.

The aim of this study was to analyze life satisfaction in a sample of 70 children and adolescents (M = 12.21, SD = 2.85) with Attention-Deficit/Hyperactivity Disorder (ADHD), according to parents' and children's/adolescents' reports. In addition, we examined the influence of a series of child/adolescent variables (ADHD presentation, and Conduct Disorder (CD) symptoms, age, gender, and pharmacological support status) on their levels of life satisfaction. Results indicated moderate correlations between children's/adolescents' and parents' perceptions of life satisfaction (r = .40; p < .01), with school being the area with the lowest levels of satisfaction. Also, 44.3% of the sample of parents reported that ADHD drastically interferes negatively in this context. Examining the effects of child/adolescent variables, only the variables age and CD symptoms generated statistically significant differences, showing that as children/adolescents grow up and/or present associate symptoms of CD, perceptions of life satisfaction tend to be more negative. These variables explained 34.5% of the variance of a composite score of life satisfaction, demonstrating a negative effect over the dependent variable. These results might have important implications for diagnosis and intervention in ADHD, as they highlight the relevance of considering life satisfaction as an important aspect to consider in both processes. Further studies must look more deeply into the mechanisms that explain these findings.

BIO bragg gratings on microfibers for label-free biosensing.

Discovering nanoscale phenomena to sense biorecognition events introduces new perspectives to exploit nanoscience and nanotechnology for bioanalytical purposes. Here we present Bio Bragg Gratings (BBGs), a novel biosensing approach that consists of diffractive structures of protein bioreceptors patterned on the surface of optical waveguides, and tailored to transduce the magnitude of biorecognition assays into the intensity of single peaks in the reflection spectrum. This work addresses the design, fabrication, and optimization of this system by both theoretical and experimental studies to explore the fundamental physicochemical parameters involved. Functional biomolecular gratings are fabricated by microcontact printing on the surface of tapered optical microfibers, and their structural features were characterized. The transduction principle is experimentally demonstrated, and its quantitative bioanalytical prospects are assessed in a representative immunoassay, based on patterned protein probes and selective IgG targets, in label-free conditions. This biosensing system involves appealing perspectives to avoid unwanted signal contributions from non-specific binding, herein investigated in human serum samples. The work also proves how the optical response of the system can be easily tuned, and it provides insights into the relevance of this feature to conceive multiplexed BBG systems capable to perform multiple label-free biorecognition assays in a single device.

Label-free detection of C-Reactive protein using bioresponsive hydrogel-based surface relief diffraction gratings.

Responsive hydrogel-based surface relief gratings have demonstrated great performances as transducers in optical sensing. However, novel and smart designs of hydrogels are needed for the appropriate detection of analytes and biomolecules since the existing materials are very limited to specific molecules. In this work, a biosensing system based on surface relief gratings made of bioresponsive hydrogels has been developed. In particular, the hydrogel contains phosphocholine moieties to specifically recognize C-Reactive protein (CRP). The CRP-Sensing hydrogel capacity to selectively detect CRP was fully demonstrated. Using Direct Laser Interference Patterning, micro-gratings were created on CRP-Sensing hydrogel substrates and applied for the label-free sensing of CRP using a simple laser-based homemade optical setup. Limits of detection (LOD) and quantification (LOQ) in human serum dilutions of 1.07 and 8.92 mg L-1, respectively, were reached. These results demonstrate that the biosensing system allows the selective label-free detection of CRP within concentration ranges around those related to risks of cardiovascular diseases and sepsis. Besides, amplification strategies have been carried out improving the sensitivity, widening the linear range, and reaching better LOD and LOQ (0.30 mg L-1 and 4.36 mg L-1). Finally, all the approaches were tested for the quantification of CRP in certified human serum with recoveries of around 100%.

[Empirically Supported Psychological Treatments for Children and Adolescents: State of the Art].

Empirically Supported Psychological Treatments for Children and Adolescents: State of the Art. BACKGROUND: The empirical evidence accumulated on the efficacy, effectiveness, and efficiency of psychotherapeutic treatments in children and adolescents calls for an update. The main goal of this paper objective was to carry out a selective review of empirically supported psychological treatments for a variety of common psychological disorders and problems in childhood and adolescence. METHOD: A review was carried out of the psychological treatments for different psychological disorders and problems in social-emotional or behavioral adjustment in the child-adolescent population according to the Spanish National Health System (Clinical Practice Guidelines) levels of evidence and degrees of recommendation. RESULTS: The findings suggest that psychological treatments have empirical support for addressing a wide range of psychological problems in these developmental stages. The degree of empirical support ranges from low to high depending on the phenomenon analyzed. The review suggests unequal progress in the different fields of intervention. CONCLUSIONS: From this update, psychologists will be able to make informed decisions when implementing those empirically supported treatments to address the problems that occur in childhood and adolescence.

Personal and Instructional Variables Related to the Learning Process in Postgraduate Courses.

BACKGROUND: The aim of the present study was to analyze the relationship between different student characteristics (gender, reason for choosing the master's degree and specialty), the instructional context (perceived quality of the instructional process), and the characteristics of the learning process (practical learning strategies and study approach) in postgraduate study. METHOD: 621 students completed the Motivated Strategies for Learning Questionnaire, the Inventory of Study Processes, the Inventory of Self-Regulatory Learning Processes and a questionnaire developed ad hoc about instructional processes. RESULTS: The data indicates notable differences between men's and women's involvement in their postgraduate study. Furthermore, the results also suggest that it is only when postgraduate courses are done for vocational reasons and students receive quality instruction that the two factors ensure students prioritize deep learning processes (e.g., significant learning, reflections on meaning, comprehensive learning) versus a surface learning approach. (e.g., memorization, lack of reflection). CONCLUSIONS: The results may be of interest for the design of postgraduate university education policy in relation to the selection processes for both students and teachers.

Multimodal Protocol for Assessing Metacognition and Self-Regulation in Adults with Learning Difficulties.

Learning disabilities (LDs) encompass disorders of those who have difficulty learning and using academic skills, exhibiting performance below expectations for their chronological age in the areas of reading, writing, and/or mathematics. Each of the disorders making up the LDs involve different deficits; however, some commonalities can be found within that heterogeneity, such in terms of learning self-regulation and metacognition. Unlike in early ages and later educational levels, there are hardly any evidence-based evaluation protocols for adults with LDs. LDs influence academic performance but also have serious consequences in professional, social, and family contexts. In response to this, the current work proposes a multimodal evaluation protocol focused on metacognitive, self-regulation of learning, and emotional processes, which make up the basis of the difficulties in adults with LDs. The assessment is carried out through analysis of the on-line learning process using a variety methods, techniques, and sensors (e.g., eye tracking, facial expressions of emotion, physiological responses, concurrent verbalizations, log files, screen recordings of human-machine interactions) and off-line methods (e.g., questionnaires, interviews, and self-report measures). This theoretically-driven and empirically-based guideline aims to provide an accurate assessment of LDs in adulthood in order to design effective prevention and intervention proposals.

Supplementation with high-content docosahexaenoic acid triglyceride in attention-deficit hyperactivity disorder: a randomized double-blind placebo-controlled trial.

Background: Attention-deficit hyperactivity disorder (ADHD) is a complex disorder in terms of etiology, clinical presentation, and treatment outcome. Pharmacological and psychological interventions are recommended as primary treatments in ADHD; however, other nonpharmacological intervention such as a dietary supplementation with omega-3 polyunsaturated fatty acids (ω-3 PUFAs) has emerged as an attractive option. Purpose: The objective of the present study was to assess whether dietary supplementation with highly concentrated ω-3 docosahexaenoic acid (DHA) triglyceride may improve symptoms in ADHD. Method: A 6-month prospective double-blind placebo-controlled randomized clinical trial was designed in 66 patients with ADHD, aged between 6 and 18 years. Participants in the experimental group received a combination of ω-3 fatty acids (DHA 1,000 mg, eicosapentaenoic acid 90 mg, and docosapentaenoic acid 150 mg). Instruments included d2-test, AULA Nesplora, EDAH scales, and abbreviated Conner's Rating Scale. Results: In the cognitive test, between-group differences were not found, but within-group differences were of a greater magnitude in the DHA group. Between-group differences in favor of the DHA arm were observed in behavioral measures, which were already detected after 3 months of treatment. Results were not changed when adjusted by ADHD medication. Conclusions: This study provides further evidence of the beneficial effect of supplementation with ω-3 DHA in the management of ADHD.

Disk-based one-dimensional photonic crystal slabs for label-free immunosensing.

One-dimensional photonic crystal slabs are periodic optical nanostructures that produce guided-mode resonance. They couple part of the incident light into the waveguide generating bandgaps in the transmittance spectrum, whose position is sensitive to refractive index variations on their surface. In this study, we present one-dimensional photonic crystal slab biosensors based on the internal nanogrooved structure of Blu-ray disks for label-free immunosensing. We demonstrated that this polycarbonate structure coated with a critical thickness of TiO2 generates guided-mode resonance. Its optical behavior was established comparing it with other compact disk structures. The results were theoretically calculated and experimentally demonstrated, all them being in agreement. The bioanalytical performance of these photonic crystals was experimentally demonstrated in a model assay to quantify IgGs as well as in two immunoassays to determine the biomarkers C-reactive protein and lactate dehydrogenase (detection limits of 0.1, 87, and 13 nM, respectively). The results are promising towards the development of new low-cost, portable, and label-free optical biosensors that join these photonic crystals with dedicated bioanalytical scanners based on compact disk drives.