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The conventional approach to developing light-sensitive glycosidase activity regulators, involving the combination of a glycomimetic moiety and a photoactive azobenzene module, results in conjugates with differences in glycosidase inhibitory activity between the interchangeable E and Z-isomers at the azo group that are generally below one-order of magnitude. In this study, we have exploited the chemical mimic character of sp 2 -iminosugars to access photoswitchable p- and o-azobenzene α-O-glycosides based on the gluco-configured representative ONJ. Notably, we achieved remarkably high switching factors for glycosidase inhibition, favoring either the E- or Z-isomer depending on the aglycone structure. Our data also indicate a correlation between the isomeric state of the azobenzene module and the selectivity towards α- and β-glucosidase isoenzymes. The most effective derivative reached over a 10 3 -fold higher inhibitory potency towards human β-glucocerebrosidase in the Z as compared with the E isomeric form. This sharp contrast is compatible with ex-vivo activation and programmed self-deactivation at physiological temperatures, positioning it as a prime candidate for pharmacological chaperone therapy in Gaucher disease. Additionally, our results illustrate that chemical tailoring enables the engineering of photocommutators with the ability to toggle inhibition between α- and β-glucosidase enzymes in a reversible manner, thus expanding the versatility and potential therapeutic applications of this approach., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

One of the current challenges in medicine is to achieve a rapid and unequivocal detection and quantification of extremely low levels of disease biomarkers in complex biological samples. Here, we present the development and analytical evaluation of a low-cost smartphone-based system designed for ultrasensitive detection of the prostate-specific antigen (PSA) using two detection alternatives: electrochemical or optical, by coupling the smartphone with a portable potentiostat or magnifying lenses. An antibody tagged with gold nanoparticles (AuNPs), and indium tin oxide coated polyethylene terephthalate platform (ITO-PET) have been used to develop a sandwich-type immunoassay. Then, a controlled silver electrodeposition on the AuNPs surface is carried out, enhancing their size greatly. Due to such strong nanoparticle-size amplification (from nm to μm), the final detection can be dual, by measuring current intensity or the number of silver-enlarged microstructures generated. The proposed strategies exhibited limit detections (LOD) of 102 and 37 fg/mL for electrochemical and optical detection respectively. The developed immunosensor reaches excellent selectivity and performance characteristics to quantify biomarkers at clinically relevant values without any pretreatment. These proposed procedures could be useful to check and verify possible recurrence after clinical treatment of tumors or even report levels of disease serum biomarkers in early stages., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Guillermo Redondo-Fernández reports financial support was provided by Spanish Ministry of Science and Innovation. Guillermo Redondo-Fernández reports financial support was provided by Principado de Asturias GRUPIN, co-financed by the Feder Programme of the European Union. Guillermo Redondo-Fernández reports financial support was provided by University of Oviedo., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

A major challenge in the 21st century is the development of point-of-care diagnostic tools capable to detect and quantify disease biomarkers in a straightforward, affordable, sensitive, and specific manner. The remarkable plasmonic properties of gold nanoparticles (AuNPs) have promoted their use for development of simple methodologies for nucleic acid detection in combination with a variety of oligonucleotides amplification techniques. Here, assemblies of AuNPs with Multicomponent Nucleic Acid enzymes (MNAzymes) has been successfully used in the design of a highly sensitive and simple bioassay for rapid spectroscopic detection and quantification of miRNA-4739 in blood samples. The miRNA selected is a doxorubicin chemoresistant biomarker in breast cancer which overexpression promotes the proliferation, progression, and survival of cancer cells. In this work, two alternatives experimental designs, based on use of MNAzymes and AuNPs, have been optimized and applied for sensitive miRNA-4739 quantification: one based on a traditional direct measurement of wavelength shift and a second non-conventional simple approach based on isolation and measurement of free nanoparticles absorbance. Improvement in sensitivity and, higher measurement accuracy and precision were achieved with the second approach. The developed bioassay provides a detection limit as low as 7 pmolL -1 for miRNA-4739 quantification and performed satisfactory selectivity and well practical applicability by analysis of the miRNA-4739 in blood, demonstrating that the proposed strategy is a promising and suitable spectroscopic method for breast cancer diagnosis thought liquid biopsy of circulating tumoral cells., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

In-depth characterization of functionalized nanomaterials is still a remaining challenge in nanobioanalytical chemistry. In this work, we propose the online coupling of Asymmetric Flow Field-Flow Fractionation (AF4) with UV/Vis, Multiangle Light Scattering (MALS) and Inductively Coupled Plasma-Tandem Mass Spectrometry (ICP-MS/MS) detectors to carry out, in less than 10 min and directly in the functionalization reaction mixture, the complete characterization of gold nanoparticles (AuNPs) functionalized with oligonucleotides and surface-modified with polyethylene glycol (PEG). AF4 separation provided full separation of the bioconjugates from the original AuNPs while P/Au and S/Au ICP-MS/MS ratios in the bioconjugate fractographic peaks could be used to compute the corresponding stoichiometries, oligonucleotide/AuNP and PEG/AuNPs. MALS detection clearly showed the coexistence of two distinct nanoparticulated populations in the bioconjugation mixture, which were demonstrated to be different not only in size but in functionality as well. The major bioconjugate population showed lower hydrodynamic ratios (18 nm) with higher and steadier oligonucleotides/AuNPs (92) and PEG/AuNPs (2350) stoichiometries, in comparison to the minor abundant population (54 nm, 51 and 1877, respectively). Moreover, the ratio between the absorbance signals measured at 520 nm and 650 nm reflects a lower AuNP aggregation in the major (10.5) than in the minor (4.5) population. Results obtained prove the benefits of a detailed characterization to find out if subsequent purification of functionalized AuNP-oligonucleotides is required to design more efficiently their final bioanalytical application., Competing Interests: Declaration of competing interest The authors declare no financial interests/personal relationships which may be considered as potential competing interests., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The marine cyanobacterium Prochlorococcus is one of the main primary producers on Earth, which can take up glucose by using the high affinity, multiphasic transporter GlcH. We report here the overexpression of glcH from Prochlorococcus marinus strain SS120 in Escherichia coli. Modeling studies of GlcH using the homologous MelB melibiose transporter from Salmonella enterica serovar Typhimurium showed high conservation at the overall fold. We observed that an important structural interaction, mediated by a strong hydrogen bond between D8 and R141, is conserved in Prochlorococcus, although the corresponding amino acids in MelB from Salmonella are different. Biased docking studies suggested that when glucose reaches the pocket of the transporter and interacts with D8 and R141, the hydrogen bond network in which these residues are involved could be disrupted, favoring a conformational change with the subsequent translocation of the glucose molecule towards the cytoplasmic region of the pmGlcH structure. Based on these theoretical predictions and on the conservation of N117 and W348 in other MelB structures, D8, N117, R141 and W348 were mutated to glycine residues. Their key role in glucose transport was evaluated by glucose uptake assays. N117G and W348G mutations led to 17 % decrease in glucose uptake, while D8G and R141G decreased the glucose transport by 66 % and 92 % respectively. Overall, our studies provide insights into the Prochlorococcus 3D-structure of GlcH, paving the way for further analysis to understand the features which are involved in the high affinity and multiphasic kinetics of this transporter., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

Objectives: To evaluate whether simplification of antiretroviral treatment to dual therapy (DT) negatively impacts immune recovery (IR), immune activation and inflammation (IA/I), and HIV reservoir., Methods: An open-label, single-centre, randomized controlled trial conducted in adult virologically suppressed HIV-infected patients on triple therapy (TT) with elvitegravir-cobicistat, emtricitabine and tenofovir alafenamide or dolutegravir (DTG), abacavir, and lamivudine (3TC). Participants were randomized to continue TT or switch to DTG, or darunavir/cobicistat (DRVc) plus 3TC. IR was assessed by CD4 + /CD8 + ratio at 48 and 96 weeks. Changes in immune activation, proliferation, exhaustion, senescence, and apoptosis in CD4 + and CD8 + T cells, plasma sCD14, hsCRP, D-dimers, β2-microglobulin, IL-6, TNF-α and IP-10 levels, cell-associated HIV-DNA (CA-DNA), and unspliced HIV-RNA (usRNA) were also analysed., Results: One hundred and fifty-one participants were enrolled. Fourteen patients did not complete the follow up. In the ITT and PP analysis, the IR was similar between the treatment arms. In the ITT analysis, the median increase in CD4 + /CD8 + ratio was 0.10, 0.04, and 0.07 at week 48, and 0.09, 0.05, and 0.08 at week 96 for TT, DTG/3TC, and DRVc/3TC, respectively. After adjusting for confounding factors, the slopes of changes in CD4 + /CD8 + ratio over time were independent of treatment (F = 1.699; p = 0.436) and related only to baseline values (F = 756.871; p = 0.000). There were no differences in IA/I, CA-DNA, or usRNA between treatment arms., Discussion: Both IR and IA/I, CA-DNA, and usRNA were similar in the three treatment groups, regardless of maintaining TT or simplifying to DTG/3TC or DRVc/3TC in virologically suppressed HIV-infected patients., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Purpose: The aim of this study was to assess racial/ethnic and socioeconomic disparities in the difference between atherosclerotic vascular disease prevalence measured by a multitask convolutional neural network (CNN) deep learning model using frontal chest radiographs (CXRs) and the prevalence reflected by administrative hierarchical condition category codes in two cohorts of patients with coronavirus disease 2019 (COVID-19)., Methods: A CNN model, previously published, was trained to predict atherosclerotic disease from ambulatory frontal CXRs. The model was then validated on two cohorts of patients with COVID-19: 814 ambulatory patients from a suburban location (presenting from March 14, 2020, to October 24, 2020, the internal ambulatory cohort) and 485 hospitalized patients from an inner-city location (hospitalized from March 14, 2020, to August 12, 2020, the external hospitalized cohort). The CNN model predictions were validated against electronic health record administrative codes in both cohorts and assessed using the area under the receiver operating characteristic curve (AUC). The CXRs from the ambulatory cohort were also reviewed by two board-certified radiologists and compared with the CNN-predicted values for the same cohort to produce a receiver operating characteristic curve and the AUC. The atherosclerosis diagnosis discrepancy, Δ vasc , referring to the difference between the predicted value and presence or absence of the vascular disease HCC categorical code, was calculated. Linear regression was performed to determine the association of Δ vasc with the covariates of age, sex, race/ethnicity, language preference, and social deprivation index. Logistic regression was used to look for an association between the presence of any hierarchical condition category codes with Δ vasc and other covariates., Results: The CNN prediction for vascular disease from frontal CXRs in the ambulatory cohort had an AUC of 0.85 (95% confidence interval, 0.82-0.89) and in the hospitalized cohort had an AUC of 0.69 (95% confidence interval, 0.64-0.75) against the electronic health record data. In the ambulatory cohort, the consensus radiologists' reading had an AUC of 0.89 (95% confidence interval, 0.86-0.92) relative to the CNN. Multivariate linear regression of Δ vasc in the ambulatory cohort demonstrated significant negative associations with non-English-language preference (β = -0.083, P < .05) and Black or Hispanic race/ethnicity (β = -0.048, P < .05) and positive associations with age (β = 0.005, P < .001) and sex (β = 0.044, P < .05). For the hospitalized cohort, age was also significant (β = 0.003, P < .01), as was social deprivation index (β = 0.002, P < .05). The Δ vasc variable (odds ratio [OR], 0.34), Black or Hispanic race/ethnicity (OR, 1.58), non-English-language preference (OR, 1.74), and site (OR, 0.22) were independent predictors of having one or more hierarchical condition category codes (P < .01 for all) in the combined patient cohort., Conclusions: A CNN model was predictive of aortic atherosclerosis in two cohorts (one ambulatory and one hospitalized) with COVID-19. The discrepancy between the CNN model and the administrative code, Δ vasc , was associated with language preference in the ambulatory cohort; in the hospitalized cohort, this discrepancy was associated with social deprivation index. The absence of administrative code(s) was associated with Δ vasc in the combined cohorts, suggesting that Δ vasc is an independent predictor of health disparities. This may suggest that biomarkers extracted from routine imaging studies and compared with electronic health record data could play a role in enhancing value-based health care for traditionally underserved or disadvantaged patients for whom barriers to care exist., (Copyright © 2021 American College of Radiology. Published by Elsevier Inc. All rights reserved.)

Mercury (Hg) is one of the most toxic environmental pollutants, and is biocondensed via the food chain. Selenium (Se) is an essential element that possesses an antagonistic property towards Hg in vivo. The antagonistic property is explained by the assumption that Hg and Se directly interact to form HgSe nanoparticles (HgSe NPs) in organs. It is presumed that the toxic effects of HgSe NPs are lower than that of ionic Hg; however, no precise evaluation has been conducted so far. In the present study, we evaluated the distribution of HgSe NPs ingested in Se-deficient rats. The recovery of serum selenoproteins from a deficient level was not observed in rats orally administered HgSe NPs. In addition, the excretion of Hg and Se via urine was not observed. Interestingly, the biosynthesis of selenoproteins and urinary selenometabolites would have required the production of selenide through the degradation of HgSe NPs. Therefore, it seems that selenide and Hg are not released from HgSe NPs in vivo. The administration of HgSe NPs did not increase Hg and Se concentrations in organs, and almost all HgSe NPs were recovered in feces, indicating no or low bioaccessibility of HgSe NPs even in Se-deficient rats. These results suggest that HgSe NPs are biologically inert and do not become a secondary environmental pollutant of Hg., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Arachidonic acid (AA) is a fatty acid involved in the modulation of several ion channels. Previously, we reported that AA activates the high conductance Ca 2+ - and voltage-dependent K + channel (BK) in vascular smooth muscle depending on the expression of the auxiliary β1 subunit. Here, using the patch-clamp technique on BK channel co-expressed with β1 subunit in a heterologous cell expression system, we analyzed whether AA modifies the three functional modules involved in the channel gating: the voltage sensor domain (VSD), the pore domain (PD), and the intracellular calcium sensor domain (CSD). We present evidence that AA activates BK channel in a direct way, inducing VSD stabilization on its active configuration observed as a significant left shift in the Q-V curve obtained from gating currents recordings. Moreover, AA facilitates the channel opening transitions when VSD are at rest, and the CSD are unoccupied. Furthermore, the activation was independent of the intracellular Ca 2+ concentration and reduced when the BK channel was co-expressed with the Y74A mutant of the β1 subunit. These results allow us to present new insigths in the mechanism by which AA modulates BK channels co-expressed with its auxiliary β1 subunit., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Hepatobiliary resections are challenging due to the complex liver anatomy. Three-dimensional printing (3DP) has gained popularity due to its ability to produce anatomical models based on the characteristics of each patient., Methods: A multicenter study was conducted on complex hepatobiliary tumours. The endpoint was to validate 3DP model accuracy from original image sources for application in the teaching, patient-communication, and planning of hepatobiliary surgery., Results: Thirty-five patients from eight centers were included. Process testing between 3DP and CT/MRI presented a considerable degree of similarity in vascular calibers (0.22 ± 1.8 mm), and distances between the tumour and vessel (0.31 ± 0.24 mm). The Dice Similarity Coefficient was 0.92, with a variation of 2%. Bland-Altman plots also demonstrated an agreement between 3DP and the surgical specimen with the distance of the resection margin (1.15 ± 1.52 mm). Professionals considered 3DP at a positive rate of 0.89 (95%CI; 0.73-0.95). According to student's distribution a higher success rate was reached with 3DP (median:0.9, IQR: 0.8-1) compared with CT/MRI or 3D digital imaging (P = 0.01)., Conclusion: 3DP hepatic models present a good correlation compared with CT/MRI and surgical pathology and they are useful for education, understanding, and surgical planning, but does not necessarily affect the surgical outcome., (Copyright © 2020 International Hepato-Pancreato-Biliary Association Inc. Published by Elsevier Ltd. All rights reserved.)

Introduction: Patients with moderate-severe cerebral palsy require the support of their caregivers to carry out the activities of daily living (ADLs)., Objectives: To describe the comorbidities, need for care in children with cerebral palsy and to analyse the influence of the degree of motor involvement, nutritional status and other neurological disorders., Methods: Cross-sectional and observational study. Patients with cerebral palsy degrees III-IV-V according to the Gross Motor Function Classification System (GMFCS) have been studied. A record of comorbidities has been made and body composition has been studied using anthropometry and bioimpedance. In addition, a caregiver burden survey on ADLs has been carried out (10 items on the different actions: hygiene, clothing, transfers, sleeping and feeding). Which variables have the greatest influence on the perception of difficulty in performing ADLs have been studied., Results: A total of 69 patients (50.7% women, mean age 10.46 ± 0.4 years) were analysed, with GMFCS grades: grade III 36.2% (N=25), grade IV 29.0% (N=20), grade V 34.8% (N=24). A relationship was found between the caregiver burden score and GMFCS grade (P=0.003) and intellectual disability (P<0.001). However, regardless of the degree of GMFCS and intellectual disability, there is greater difficulty in performing ADLs in relation to lower values in weight (Z-score) (P=0.028), fat mass (kg) (P=0.035), fat mass (%) (P=0.094), body mass index (Z-score) (P=0.086)., Conclusions: In addition to the degree of clinical impairment, nutritional status is a factor that influences the caregiver's difficulty in performing the ADLs in cerebral palsy patients on which we can act to improve this problem., (Copyright © 2021 Asociación Española de Pediatría. Publicado por Elsevier España, S.L.U. All rights reserved.)

Background: High alcohol consumption and alcohol dependence are only partly genetically correlated and they differ considerably in their correlations with other traits. The existence of genetic correlation among alcohol dependence and psychiatric disorders may be attributed to the presence of a general psychopathology factor, the p factor. This study investigates the relationship of polygenic risk to general psychopathology and to high alcohol consumption on alcohol dependence., Methods: Participants were 524 alcohol-dependent patients and 729 controls. Polygenic risk scores (PRS) were computed for alcohol consumption (drinks per week) and nine psychiatric disorders. Principal component analysis (PCA) applied to the psychiatric PRS was used to calculate the first principal component as a proxy of the polygenic p factor., Results: Both the polygenic p factor and the drinks per week PRS were associated with alcohol dependence in our sample. Both variables are only weakly correlated, contributing additively to the risk for alcohol dependence. Sensitivity analyses showed that the polygenic p factor was also associated with alcohol dependence in the subset of patients without any psychiatric or substance use comorbidity., Conclusions: Polygenic risk for alcohol dependence can be split at least into two components, involved in general psychopathology and high alcohol consumption. The first component of PCA based on PRS for different psychiatric disorders allows estimation of the contribution of the polygenic p factor to alcohol dependence. The pleiotropic effects of genetic variants across psychiatric disorders are mainly manifested as alcohol dependence in some patients., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Introduction: Outcomes in patients diagnosed of acute lymphoblastic leukemia with Philadelphia chromosome (Ph-ALL) remains unfavourable compared to other subtypes of acute lymphoblastic leukemia despite improvements in drug treatments as well as advances in hematopoietic stem cell transplantation (HSCT)., Patients and Methods: The role of allogeneic HSCT in Ph-ALL patients has been analysed through a multicentric study where data belonging to 70 patients diagnosed of this entity in different center that received HSCT between years 1998 and 2014, were reported by the Grupo Español de Trasplante Hematopoyético (GETH)., Results: The performance of HSCT from year 2004, in first complete remission (CR) status with thymoglobulin (ATG) based conditioning had a favorable impact on overall survival (OS). HSTC performance from year 2004, in first CR with ATG-based conditioning in addition to acute graft versus host disease (aGvHD) development, increased event free survival (EFS). Treatment with imatinib as well as undetectable minimal residual disease (MRD) prior to HSCT, combined with aGvHD, reduced risk of relapse (RR). Patient age less than 10 years when HSCT, first CR and ATG-based conditioning were associated to a lower transplant related mortality (TRM)., Conclusions: Patients that could achieve first CR that also received ATG-based conditioning had a better OS and EFS, so HSCT should be considered for this group of patients., (Copyright © 2021. Publicado por Elsevier España, S.L.U.)

Objective: The International Commission on Radiological Protection (ICRP) has highlighted the large number of medical specialties using fluoroscopy outside imaging departments without programmes of radiation protection (RP) for patients and staff. Vascular surgery is one of these specialties and endovascular aneurysm repair (EVAR) is one of the most challenging procedures requiring RP guidance and optimisation actions. The recent European Directive on Basic Safety Standards requires the use and regular update of diagnostic reference levels (DRL) for interventional procedures. The objective of the study was to know the doses of patients undergoing EVAR with mobile Xray systems and with hybrid rooms (fixed Xray systems), to obtain national DRLs and suggest optimisation actions., Methods: The Spanish Chapter of Endovascular Surgery launched a national survey that involved hospitals for 10 autonomous communities representing the 77% of the Spanish population (46.7 million inhabitants). Patient dose values from mobile Xray systems were available from nine hospitals (sample of 165 EVAR procedures) and data from hybrid rooms, from seven hospitals, with dosimetric data from 123 procedures. The initial national DRLs have been obtained, as the third quartile of the median values from the different centres involved in the survey., Results: The proposed national DRLs are 278 Gy cm 2 for hybrid rooms and 87 Gy cm 2 for mobile Xray systems, and for cumulative air kerma (cumulative AK) at the patient entrance reference point, 1403 mGy for hybrid rooms, and 292 mGy for mobile systems., Conclusion: An audit of patient doses for EVAR procedures to identify optimised imaging protocol strategies is needed. It is also appropriate to evaluate the diagnostic information required for EVAR procedures. The increase by a factor of 3.2 (for kerma area product) and 4.8 (for cumulative AK) in the DRLs needs to be justified when the procedures are performed in the hybrid rooms rather than with mobile Xray systems., (Copyright © 2020 European Society for Vascular Surgery. Published by Elsevier B.V. All rights reserved.)

SARS-CoV-2 pandemic dimension has affected the Hospital Pediatrics Medicine assistance in our country. New challenges generated by COVID-19 require a series of proactive measures, based on existing scientific knowledge and standards of good practice, that allow the Pediatric Hospital services readiness and operability. Hospital Internal Pediatrics, as responsible of integral care of the hospitalized child, plays a leading role in the new hospital model emerging from this crisis. This review analyzes the impact of the current SARS-CoV-2 epidemic on pediatric care, and perspective of new COVID-19 outbreaks in coexistence with other viral infections. Changes secondary to pandemic involved in Hospital Pediatric units, how to prepare for future epidemics, also the involvement of pediatric units in adult care and the possible opportunities for improvement need to be revised. Assistance of patients with chronic complex conditions in epidemic circumstances, safety aspects, opportunities for teaching and ethical considerations are reviewed. The Spanish Society of Hospital Pediatrics Medicine offers with this article a series of resources for Internal Pediatric Medicine practitioners responsible to face next challenges in pediatric hospitalization units., (Copyright © 2020 Asociación Española de Pediatría. Publicado por Elsevier España, S.L.U. All rights reserved.)

Introduction: Community-Acquired Pneumonia (CAP) is one of the most frequent causes of hospital admission in children. Our objective is to measure the impact of the introduction of pneumococcal conjugate vaccines on the hospitalization of previously healthy children due to CAP., Method: From 2011 to 2016, a partially retrospective, prospective, and descriptive study was carried out on healthy pediatric patients (3 months-14 years old) with CAP, who required hospital admission. Clinical, epidemiological, and demographic characteristics were collected, and vaccination status was obtained from medical records., Results: A total of 292 cases were included, with a mean age of 33.4 months, 54% males. There was a progressive and significant 42% decrease in the number of admissions each year, without significant changes in the annual percentage of parapneumonic pleural effusion (PPE). Fifty-six percent of patients were immunized with a pneumococcal conjugate vaccine (PCV). The percentage of children who were not vaccinated decreased by 14%, and the coverage with PCV-13 increased by 46%. This revealed a significant increase of PPE in vaccinated patients with PCV-7 (63%) compared with unvaccinated (45%) and with PCV-13 (57%), without association with the presence of severe PPE. Moreover, no significant differences in severity or hospital stay were observed in unvaccinated patients, compared to those who were vaccinated. In >2-year-olds, we observed a significant increase in PPE (59%) compared to 45% in younger children., Conclusions: The increase in vaccination coverage with PCV-13 resulted in a decrease in hospitalizations due to CAP and PPE. Vaccination with PCV-7 is associated in our sample with an increase in PPE but not with severe PPE nor an increase in the hospital stay. There was an epidemiological shift of severe forms of pneumonia and empyema at later ages (>2 years)., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

A 32 base pair deletion in the C-C chemokine receptor type gene (CCR5-Δ32), the main Human Immunodeficiency Virus (HIV) co-receptor results in a non-functional protein. Individuals homozygous for the CCR5-Δ32 mutation are resistant to HIV infection. Here we report the generation, from pro-erythroblast enriched Peripheral Blood Mononuclear Cells (PBMCs) from a naturally occurring CCR5-Δ32/Δ32 individual, of the fully characterized iPSC line IMEDEAi008-A. The new line has normal karyotype, carry the Δ32 mutation in homozygosity, is free of plasmid integrations, express high levels of pluripotency markers and can differentiate into all three germ layers., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier B.V.)

We present here one of the first cases of virological failure during treatment with bictegravir/emtricitabine/tenofovir alafenamide (BIC/FTC/TAF). On March 2019, an antiretroviral-experienced HIV-infected patient was admitted to hospital because of cerebral toxoplasmosis. After undergoing treatment with sulfadiazine-pyrimethamine for two weeks, the patient initiated a BIC/FTC/TAF treatment, with 6.01 HIV RNA Log copies/mL, and 37 CD4 cells/μL. After two months under antiretroviral therapy (ART), acute neurologic deterioration with epilepsy, right hemiparesis and dysphagia occurred, leading to nasogastric nutrition and treatment. After several weeks, virological failure was confirmed with 4.01 HIV RNA Log copies/mL and R263K and M184V resistance mutations were detected., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020. Published by Elsevier B.V.)

Preparation for oxidative stress (POS), i.e., the buildup of endogenous antioxidants during metabolic depression or low oxygen stress conditions, has been observed in at least 8 animal phyla under controlled conditions in laboratory. Despite the expected implications on the endurance to extreme environments and ecosystem occupation, the extent to which POS occurs in animals under natural conditions remains unexplored. Therefore, we took advantage of the natural history of the Brazilian Caatinga's frog Proceratophrys cristiceps to investigate the modulation of endogenous antioxidants and redox balance in their skeletal muscle and to verify if POS occurs under natural conditions. Expectedly, estivating frogs had low levels of the oxidative metabolism enzymes. Citrate synthase and isocitrate dehydrogenase activities were 36% and 25% lower than those in active frogs respectively. We found an overall upregulation of antioxidants in estivating P. cristiceps. Reduced glutathione concentration was 61% higher in estivating frogs than that in active animals. During estivation the activities of the hydroperoxide detoxification enzymes catalase, glutathione peroxidase, and glutathione H 2 O 2 -peroxidase were 48%, 57%, and 78% greater than those during the rainy season. Moreover, estivating frogs had a 47% lower ratio of disulfide to total glutathione levels than active frogs. Our findings confirm the occurrence of 'preparation for oxidative stress' in naturally estivating frogs and paves the way for further research on the redox biology of animals under natural settings. Such approach might reveal biochemical strategies under ecologically relevant scenarios., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Introduction: Cerebral palsy (CP) is the most frequent cause of motor disability in the paediatric age. The aim of this article is the study of the nutritional status of patients with CP followed-up in a reference hospital, as well as the relationship between neurological and nutritional state., Material and Methods: A cross-sectional, observational, descriptive and analytical study was conducted on a sample consisting of 4-15years old patients with CP with Gross Motor Function Classification System (GMFCS) gradesIII-IV-V, from a specialised paediatric hospital reference area. An interview (collection of general data, medications and nutritional habits), anthropometric study and bioimpedance (BIA) measurements were carried out., Results: The study included 69 patients (recruitment 84.15%), with a mean age of 10.46±0.43years, and 50.7% females. The distribution according to GMFCS scale was: gradeIII (36.2%), gradeIV (29%), and gradeV (34.8%). According to weight for height: moderate malnutrition 21.8% (gradeV: 33.3%), severe malnutrition 5.8% (gradeV: 12.6%), overweight/obesity 23.2% (gradeIII: 24%, gradeIV: 35%). Adequate level of lean mass for height: gradeIII (36%), gradeIV (55%), and gradeV (16.7%). Fat excess: gradeIII (36%), gradeIV (40%), and gradeV (29.2%). Fat mass comparison: BIA 6.89±0.64kg versus anthropometry 5.56±4.43kg., Conclusions: In CP grade GMFCSV, the weight deficit associated with a decrease in lean body mass is common. Patients with CP grades GMFCSIII-IV have a significant prevalence of overweight/obesity. Anthropometry is a useful tool for nutritional assessment in children with CP, although fat levels could be underestimated., (Copyright © 2019. Publicado por Elsevier España, S.L.U.)

The asymmetric flow field-flow fractionation (AF4) coupled on-line with elemental (inductively coupled plasma-mass spectrometry, ICP-MS) and molecular (fluorescence and UV) detection has been investigated as a powerful tool for the characterization of bioinorganic nano-conjugates. In this study, we described methods for the characterization of biotin-antibody complexes bioconjugated with streptavidin quantum dots (QDs-SA-b-Ab). Operating parameters of AF4 separation technique were optimized and two procedures are proposed using a channel thickness of 350 μm and 500 μm. The use of a 500 μm spacer allowed to achieve an efficient AF4 separation of the QDs-SA-b-Ab complexes from the excess of individual species used in the bioconjugation that was required for a proper characterization of the bioconjugates. Optimization of the AF4 allowed a separation resolution good enough to isolate the QDs-SA-b-Ab bioconjugates from the free excess of b-Ab and QD-SA. The efficiency of the bioconjugation process could be then calculated, obtaining a value of 86% for a 1 QDs-SA: 5 b-Ab bioconjugation ratio. In addition, sample recovery around 90% was achieved., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: The curvature of the aortic arch is associated with the risk of endoleak formation after thoracic endovascular aortic repair (TEVAR). However, the adequate assessment of the angles of the aorta continues to represent a major difficulty. We developed a new program based on three-dimensional (3D) reconstructions of computed tomography (CT) scans to objectively identify the location of the aortic points of maximum curvature, and to automatically calculate the main aortic arch angles, comparing final values with visual assessment methods., Methods: This is a cross-sectional validation study of a convenience sample of subjects with multislice CT angiography scans of the thoracic aorta from an institutional imaging database. The center lumen line (CLL) of the aorta was determined semi-automatically using Endosize software. The points of maximum curvature on the CLL were determined by two methods: visually by two physicians and through a custom program., Results: The study enrolled 9 subjects: 4 with thoracic aneurysms and 5 with normal aortas. The inter-observer and inter-method correlation, agreement and reliability for each of the 3D spatial coordinates of the points of maximum curvature were appropriate. However, the aortic angles determined by visual assessment showed a very low to moderate correlation and reliability with those determined by our custom program., Conclusion: An automated custom program can reflect clinician's intuitive assessment of the location of points of maximum curvature and translate it into aortic angles with an apparently higher precision, reducing potential error and user time., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The sensitive monitoring of mercury (II) selenide nanoparticles (HgSe NPs) is of great potential relevance in environmental studies, since such NPs are believed to be the ultimate metabolic product of the lifesaving mechanism pathway of Hg detoxification in biological systems. In this context, we take advantage of using gold-nanostructured screen-printed carbon electrodes (SPCE-Au) for the rapid, simple and sensitive electrochemical quantification of engineered water-stable HgSe NPs, as an advantageous alternative to conventional elemental analysis techniques. HgSe NPs are first treated in an optimized oxidative/acidic medium for Hg 2+ release, followed by sensitive electrochemical detection by anodic stripping voltammetry (ASV). To the best of our knowledge, this is the first time that water-stable HgSe NPs are quantified using electrochemical techniques. The low limit of detection achieved (3.86 × 10 7 HgSe NPs/mL) together with the excellent repeatability (RSD: 3%), reproducibility (RSD: 5%) and trueness (relative error: 10%), the good performance in real sea water samples (recoveries of the analytical signal higher than 90%) and the simplicity/low cost of analysis make our method an ideal candidate for HgSe NPs monitoring in future environmental studies., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The Tröllaskagi Peninsula in northern Iceland hosts more than a hundred small glaciers that have left a rich terrestrial record of Holocene climatic fluctuations in their forelands. Traditionally, it has been assumed that most of the Tröllaskagi glaciers reached their Late Holocene maximum extent during the Little Ice Age (LIA). However, there is evidence of slightly more advanced pre-LIA positions. LIA moraines from Iceland have been primary dated mostly through lichenometric dating, but the limitations of this technique do not allow dating of glacial advances prior to the 18th or 19th centuries. The application of 36 Cl Cosmic-Ray Exposure (CRE) dating to Tungnahryggsjökull moraine sequences in Vesturdalur and Austurdalur (central Tröllaskagi) has revealed a number of pre-LIA glacial advances at ~400 and ~700 CE, and a number of LIA advances in the 15 th and 17 th centuries, the earliest LIA advances dated so far in Tröllaskagi. This technique hence shows that the LIA chronology in Tröllaskagi agrees with that of other European areas such as the Alps or the Mediterranean mountains. The combined use of lichenometric dating, aerial photographs, satellite images and fieldwork shows that the regional colonization lag of the commonly used lichen species Rhizocarpon geographicum is longer than previously assumed. For exploratory purposes, an alternative lichen species (Porpidia soredizodes) has been tested for lichenometric dating, estimating a tentative growth rate of 0.737 mm yr -1 ., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The mutation E280A in PSEN1 (presenilin-1) is the most common cause of early-onset familial Alzheimer's Disease (fAD). It presents autosomal dominant inheritance and frequently leads to the manifestation of the disease in relatively young individuals. Here we report the generation of one PSEN1 E280A iPSC line derived from an early-onset patient. OriP/EBNA1-based episomal plasmids containing OCT3/4, SOX2, KLF4, L-MYC, LIN28, BCL-xL and shp53 were used to reprogram oral mucosa fibroblasts. The iPSC line generated has normal karyotype, carry the E280A mutation, is free of plasmid integration, express high levels of pluripotency markers and can differentiate into all three germ layers., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Chicken anaemia virus (CAV) causes chicken infectious anaemia, a severe disease characterized by anaemia and immunosuppression and leading to serious economic losses in the poultry industry worldwide. Although CAV infection has been investigated under experimental conditions, information regarding natural infection is scarce. This report describes an outbreak of CAV infection in 18-day-old broiler chickens and investigates virus tropism in affected birds. Thymic atrophy, pale bone marrow, swelling of the legs and foot ulcers (gangrenous dermatitis) were the most common gross lesions. Severe lymphoid cell depletion in the thymic cortex and presence of intranuclear acidophilic inclusion bodies, depletion of haemopoietic cells in bone marrow and presence of lymphoid infiltrates in several organs were also observed. Immunohistochemical labelling demonstrated the CAV antigens VP1 and VP3 in several organs. The expression of both proteins was similar in the thymic cortex and in the bone marrow, the main target organs of CAV; however, VP3 expression was more abundant in the other organs. Labelling of serial sections showed that CD3 + T lymphocytes might be responsible for the dissemination of the virus from the thymus and bone marrow to other organs and that virus-induced apoptosis, mediated through caspase-3, occurred mainly in the thymus and bone marrow., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Near infrared (NIR) emitting Ag 2 S quantum dots have been synthesized, characterized and evaluated for chemical sensing applications. After their optical characterization, it was observed that the Ag 2 S quantum dots present both, excitation and emission in the NIR region, and an excellent quantum yield of 33.2%. These features are of great value for many biological applications, since autofluorescence of biological tissues or cells is minimized, and also for environmental applications, where other fluorescent concomitant species with excitation and emission in the ultraviolet-visible region might be present. Different purification procedures were evaluated in order to obtain a stable and homogeneous population of nanoparticles, which is necessary to perform quantitative analysis (e.g.: mass spectrometry-based applications), as well as to obtain a narrow NIR emission spectrum for optical applications. Comprehensive characterization using X-ray diffraction, transmission electron microscopy, and asymmetric flow field flow fractionation coupled to inductively coupled plasma-mass spectrometry has been performed to obtain parameters not easily achieved and of great interest in different research areas, such as the nanoparticle concentration NIR-emitting nanoparticles, and the surface ligand density, which directly affects to the interactions of the nanoparticles with their close environment, including unspecific adsorptions, cellular uptake, macrophage interaction, etc. Finally, the capability for sensing analytes of environmental interest based on direct-interactions of a reactive compound with the surface of the nanoparticle has been also evaluated. Quenching of the NIR emission upon interaction of the Ag 2 S quantum dots with cyanide ions was observed. Hence, a rapid, selective and highly sensitive methodology was developed for the detection of cyanide in natural waters., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Inorganic nanoparticles are a fascinating class of materials which promise great potential in numerous fields, including optical (bio)sensing. Many different kinds of such nanoparticles have been widely used for fluorescent sensing and imaging due to the different merits of fluorescent nanoparticles compared to molecular fluorophores. Progress made in the rational design of nanomaterials also allowed the synthesis of hybrid phosphorescent nanoparticles, that finds growing applications in sensing due to the combination of the interesting size- and shape-dependent properties of nanomaterials with a phosphorescence-type emission. In this review, we intend to highlight some of progress made in this active research area and update the database of various phosphorescent nanoparticles-based sensors on the basis of different sensing targets of interest in environmental, industrial and biomedical areas. Following an introduction and a discussion of merits of the synergy between nanomaterials and phosphorescence detection as compared to molecular luminophores the article assesses the kinds and specific features of nanomaterials often used in phosphorescence sensing. Specific examples on the use of phosphorescence nanoparticles in chemical sensing and bioimaging are given next. A final section intends to provide an overview of the prospects of such type of nanomaterials in the design of future devices for analytical chemistry., (Copyright © 2018. Published by Elsevier B.V.)

Mucopolysaccharydosis IIIB is the second most frequent form of Sanfilippo syndrome, a degenerative, pediatric lysosomal storage disease (LSD) characterized by severe neurological disorders and death. We have generated two iPSCs lines derived from dermal fibroblast from a MPSIIIB homozygous (P358L) donor. Cells were reprogrammed with OriP/EBNA1-based episomal plasmids containing: OCT3/4, SOX2, KLF4, L-MYC, LIN28, BCL-xL and shp53. Both cell lines are homozygous for the P358L mutation of the α-N-acetylglucosaminidase (NAGLU) gene, have normal karyotype, are free of plasmid integration, express high levels of pluripotency-associated markers and can differentiate into the three germ layers. RESOURCE TABLE: RESOURCE UTILITY: Although the generation of iPSCs has been reported for some lysosomal storage diseases (LSD) in general, and from other mutations of the NAGLU gene in particular (Lemonnier et al., 2011), this is the first time that NAGLU Pro358Leu MPSIIIB-iPSCs lines have been generated and fully characterized demonstrating their quality as iPS cells. RESOURCE DETAILS: Mucopolysaccharidosis IIIB (MPSIII, Sanfilippo syndrome type B) is a pediatric neurodegenerative disorder caused by a deficiency in NAGLU, an enzyme required for lysosomal degradation of heparin sulphate (HS). When the enzyme is absent or malfunctioning, HS accumulates in the cells of several tissues, with devastating effects in the brain and central nervous system. MPSIIIB is inherited in an autosomal recessive manner and presents an incidence between 0.03 and 0.78 cases per 1 × 10 5 live births (Fedele, 2015) depending on the country. Currently there is no therapy available. The NAGLU gene was identified in 1996, is located on chromosome 17q21.1 and contains 6 exons. More than 150 NAGLU mutations have been reported, being most of them missense (Valstar et al., 2010). All of them lead to MPSIIIB but, unlike MPSIIIA, none is predominant. The two iPSCs lines described in this report, IMEDEAi005-A and IMEDEAi005-B, (See Table 1) were generated from skin fibroblast obtained from a clinically affected homozygous donor. The mutant allele consists on a C > T transversion at nucleotide 1073 (1073 > T) resulting in a substitution of leucine for proline at codon 358 (Pro358Leu). Skin fibroblasts were reprogrammed to iPSCs by nucleofection with four OriP/EBNA1 (Epstein-Barr nuclear antigen-1) based episomal plasmids encoding 5 reprogramming genes (OCT3/4, SOX2, KLF4, L-Myc, LIN28 and BCL-xL), in addition to a short hairpin RNA against p53. The iPSCs lines showed morphology (Fig. 1A) and growth behaviour typical of human Embryonic Stem Cells (hESC), as well as normal female karyotype (46, XX) (Fig. 1B). After 12 passages, PCR analysis confirmed that both iPSCs lines had completely lost the episomal vectors (Fig. 1C). The identity of iPS cells and their parental fibroblasts was confirmed by STR analysis (Table 2, data not shown) in addition to the identification of the disease-associated mutation in the NAGLU gene by DNA sequencing (Fig. 1D). Regarding the iPSC phenotype, both lines expressed the pluripotency-associated markers: OCT3/4, NANOG, SOX2 and TRA-1-60 (Fig. 1E), and TRA-1-81 quantified by flow cytometry (Fig. 1G), resulting in 88.17% and 83.4% of TRA-1-81 positive cells in IMEDEAi005-A and IMEDEAi005-B respectively. Finally, the differentiation capacity of iPSCs lines was analyzed by embryoid body (EBs) formation. Expression of markers specific of the three germ layers was observed after at least 10 days of spontaneous differentiation (Fig. 1F). Mycoplasma analysis was negative for both iPSCs lines (Supplementary Fig. S1). Skin fibroblasts were reprogrammed to iPSCs by nucleofection with four OriP/EBNA1 (Epstein-Barr nuclear antigen-1) based episomal plasmids encoding 5 reprogramming genes (OCT3/4, SOX2, KLF4, L-Myc, LIN28 and BCL-xL), in addition to a short hairpin RNA against p53. The iPSCs lines showed morphology (Fig. 1A) and growth behaviour typical of human Embryonic Stem Cells (hESC), as well as normal female karyotype (46, XX) (Fig. 1B). After 12 passages, PCR analysis confirmed that both iPSCs lines had completely lost the episomal vectors (Fig. 1C). The identity of iPS cells and their parental fibroblasts was confirmed by STR analysis (Table 2, data not shown) in addition to the identification of the disease-associated mutation in the NAGLU gene by DNA sequencing (Fig. 1D). Regarding the iPSC phenotype, both lines expressed the pluripotency-associated markers: OCT3/4, NANOG, SOX2 and TRA-1-60 (Fig. 1E), and TRA-1-81 quantified by flow cytometry (Fig. 1G), resulting in 88.17% and 83.4% of TRA-1-81 positive cells in IMEDEAi005-A and IMEDEAi005-B respectively. Finally, the differentiation capacity of iPSCs lines was analyzed by embryoid body (EBs) formation. Expression of markers specific of the three germ layers was observed after at least 10 days of spontaneous differentiation (Fig. 1F). Mycoplasma analysis was negative for both iPSCs lines (Supplementary Fig. S1). In conclusion, we have successfully generated and characterized, for the first time to our knowledge, two human iPSCs lines from a MPSIIIB donor homozygous for the P358L NAGLU mutation. The new lines will complement the existing murine MPS IIIB model, with their potential to be used in a development of a purely human in vitro model of the disease., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

Introduction: At present, there are few validated scoring tests for assessing acute bronchiolitis (AB) severity, and limited information on their test power. The aim of the present study is to evaluate the validity of an acute bronchiolitis severity score (ABSS) to help in deciding PICU admission., Patients and Method: Prospective, descriptive, observational study of previously healthy infants under 1 year of age with AB, where the ABSS was used to compare severity as regards the need for PICU admission. The sample size was estimated as at least 175 patients. The research team was trained in the use of ABSS. All patients in the study were evaluated with ABSS daily, as well as in the case of clinical deterioration. The initial and maximum ABSS scores were contrasted to the need for PICU admission. A receiver operative curve was constructed, and the area under the curve was calculated, and the optimum point of sensitivity / specificity was estimated., Results: The study included a total of 190 patients (male / female: 58% / 42%). PICU was required in 11 (6%). The mean± SD ABSS-maximal score for patients who required and did not require PICU was 10.55± 1.12 and 6.35± 2.3, respectively (P<.001). The AUC for ABSS-maximal was 0.94 (P<.001, 95% CI: 0.90-0.98). The optimal cut-off point was set at ≥10 points for a sensitivity of 82% and a specificity of 91%., Conclusions: ABSS estimates the severity of AB regarding the need for PICU admission, with a sensitivity and specificity of clinical usefulness., (Copyright © 2017 Asociación Española de Pediatría. Publicado por Elsevier España, S.L.U. All rights reserved.)

Introduction: Acute bronchiolitis (AB) of the infant has a serious outcome in 6-16% of the hospital admitted cases. Its pathogenesis and evolution is related to the response of the T lymphocytes. The objective of the present study is to determine if the lower systemic lymphocytic response is related to a worse outcome of AB in hospitalised infants., Patients and Method: Retrospective observational-analytical study of cases-controls nested in a cohort of patients admitted due to RSV-AB between the period from October 2010 to March 2015. Those with a full blood count in the first 48hours of respiratory distress were included. Infants with underlying disease, bacterial superinfection, and premature infants <32 weeks of gestation were excluded. The main dichotomous variable was PICU admission. Other variables were: gender, age, post-menstrual age, gestational and post-natal tobacco exposure, admission month, type of lactation, and days of onset of respiratory distress. Lymphocyte counts were categorised by quartiles. Bivariate analysis was performed with the main variable and then by logistic regression to analyse confounding factors., Results: The study included 252 infants, of whom 6.6% (17) required PICU admission. The difference in mean±SD of lymphocytes for patients admitted to and not admitted to PICU was 4,044±1755 and 5,035±1786, respectively (Student-t test, P<.05). An association was found between PICU admission and lymphocyte count <3700/ml (Chi-squared, P=.019; OR: 3.2) and it was found to be maintained in the logistic regression, regardless of age and all other studied factors (Wald 4.191 P=.041, OR: 3.8)., Conclusions: A relationship was found between lymphocytosis <3700/ml in the first days of respiratory distress and a worse outcome in previously healthy infants <12 months and gestational age greater than 32 weeks with RSV-AB., (Copyright © 2017 Asociación Española de Pediatría. Publicado por Elsevier España, S.L.U. All rights reserved.)

Gene delivery is critical for the development of nucleic acid-based therapies against a range of severe diseases. The conception of non-viral (semi)synthetic vectors with low cytotoxicity and virus-like efficiency is gathering a lot of efforts, but it represents a fantastic challenge still far from accomplishment. Carbohydrate-based scaffolds offer interesting features towards this end, such as easy availability, relatively cheap cost, tuning properties and a good biocompatibility. The lack of analytical methods providing quantitative and qualitative data on their binding properties with oligonucleotides (DNA/RNA), with a minimal time and sample consumption, represents a limitation for these channels. Here, we attempted to fill the gap by hyphenation of capillary electrophoresis with mass spectrometry (CE-MS). This coupling strategy allows discriminating free and complexed DNA oligomers with cationic cyclodextrins (CDs), determining the stoichiometry where the highest observed is always DNA n : n/3(CD), and unambiguously assigning the partners through m/z detection. Very reliable data were obtained with migration time within 5.5 (standard deviation < 0.5%) and 25 min (standard deviation < 1.1%) for UV and MS detection, respectively. Furthermore, varying the nitrogen/phosphorus ratio (N/P), key parameters relating to the thermodynamics e.g. the micro and macroscopic dissociation constants K d and K D , respectively (both in low μM range) and the Gibbs free energy ΔG (-16.3 to -26.9 kJ mol -1 ), and also the cooperativity as Hill number (nH between 0.98 and 15.75) of the supramolecular process can be delineated, providing a unique tool for the high throughput screening and selection of efficient gene delivery carriers., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Introduction: The presence of apnoea in acute bronchiolitis (AB) varies between 1.2% and 28.8%, depending on the series, and is one of its most fearsome complications. The aim of this study is to determine the incidence of apnoea in hospitalised patients diagnosed with AB, and to define their associated risk factors in order to construct a prediction model., Patients and Method: A retrospective observational study of patients admitted to a tertiary hospital in the last 5 years with a diagnosis of AB, according to the classic criteria. Data was collected on the frequency of apnoea and related clinical variables to find risk factors in a binary logistic regression model for the prediction of apnoea. A ROC curve was developed with the model., Results: Apnoea was recorded during the admission of 53 (4.4%) patients out of a total 1,197 cases found. The risk factors included in the equation were: Female (OR 0.6, 95% CI: 0.27-1.37), Caesarean delivery (OR: 3.44, 95% CI: 1.5-7.7), Postmenstrual age ≤43 weeks (OR: 6.62, 95% CI: 2.38-18.7), Fever (OR: 0.33, 95% CI: 0.09-1.97), Low birth weight (OR: 5.93, 95% CI: 2.23-7.67), Apnoea observed by caregivers before admission (OR: 5.93, 95% CI: 2.64-13.3), and severe bacterial infection (OR: 3.98, 95% CI: 1.68-9.46). The optimal sensitivity and specificity of the model in the ROC curve was 0.842 and 0.846, respectively (P<.001)., Conclusions: The incidence of apnoea during admission was 4.4 per 100 admissions of AB and year. The estimated prediction model equation may be of help to the clinician in order to classify patients with increased risk of apnoea during admission due to AB., (Copyright © 2017 Asociación Española de Pediatría. Publicado por Elsevier España, S.L.U. All rights reserved.)

IL-9 is present in psoriatic lesions and is produced by lymphocytes. However, it is not known whether this cytokine is induced by relevant pathogenic triggers of psoriasis, such as Streptococcus pyogenes. Here we addressed the production of IL-9 in response to various pathogens in a psoriatic ex vivo model. Extracts of S. pyogenes and Candida albicans triggered the production of IL-9 and also IL-17A and IFN-γ. This induction was dependent on the interaction between CLA + T cells and epidermal cells. Neutralization of IL-9 reduced S. pyogenes-induced IL-17A production by CLA + T cells but had no effect on IFN-γ production. Also, IL-9 increased the survival of circulating psoriatic CLA + T cells. Co-cultures from patients with guttate or plaque psoriasis with S. pyogenes produced similar amounts of IL-9. High cytokine responses in streptococcal-driven guttate patients paralleled peaks in Psoriasis Area Severity Index and anti-streptolysin O levels. Our results confirm that IL-9 promotes inflammation in psoriasis by up-regulating IL-17A production and support the clinical association of the immune response by streptococcal-sensitized CLA + T cells with this cytokine, especially in guttate psoriasis., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)

Coupling of asymmetric flow field-flow fractionation (AF4) to an on-line elemental detection (inductively coupled plasma-mass spectrometry, ICP-MS) has been recently proposed as a powerful diagnostic tool for characterization of the bioconjugation of CdSe/ZnS core-shell Quantum Dots (QDs) to antibodies. Such approach has been used herein to demonstrate that cap exchange of the native hydrophobic shell of core/shell QDs with the bidentate dihydrolipoic acid ligands directly removes completely the eventual side nanoparticulated populations generated during simple one-pot synthesis, which can ruin the subsequent final bioapplication. The critical assessment of the chemical and physical purity of the surface-modified QDs achieved allows to explain the transmission electron microscopy findings obtained for the different nanoparticle surface modification assayed., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Herein, high-quality Mn-doped ZnS quantum dots (QDs) have been synthesized using a facile approach directly in aqueous media. The surface of the obtained QDs was further modified by cap-exchange of the native cysteine shell with dihydrolipoic acid (DHLA) ligands resulting in nanocrystals with high water-stability having an intense phosphorescent signal. Covalent bioconjugation of the DHLA-coated nanoparticles with an anti-IgG antibody was then carried out. Interestingly the QD immunoprobe (QD-labelled antibodies) maintained an intense phosphorescence emission, without any significant spectral-shift (as compared to the free QDs). Coupling of an asymmetric flow field flow fractionation technique to an elemental mass spectrometry detection enabled the accurate determination of the efficiency of the bioconjugation reaction. The obtained nanoparticle-antibody bioconjugate was then applied to develop a quantitative sandwich-type phosphorescent immunoassay for Prostate Specific Antigen (PSA), and a limit of detection (LOD) of 17 pg mL -1 of PSA was achieved and allow to quantify such biomarker in samples within clinically relevant levels. Finally, the assay was validated for the quantification of PSA in the cellular media of prostate cancer cells. Obtained results proved the robustness of the proposed immunoassay based on long-lived phosphorescence measurements against eventual photoluminescent interferences significantly affecting the conventional short-lived fluorescence detection., (Copyright © 2017. Published by Elsevier B.V.)

The self-assembling processes underlining the capabilities of facially differentiated ("Janus") polycationic amphiphilic cyclodextrins (paCDs) as non-viral gene nanocarriers have been investigated by a pluridisciplinary approach. Three representative Janus paCDs bearing a common tetradecahexanoyl multitail domain at the secondary face and differing in the topology of the cluster of amino groups at the primary side were selected for this study. All of them compact pEGFP-C3 plasmid DNA and promote transfection in HeLa and MCF-7 cells, both in absence and in presence of human serum. The electrochemical and structural characteristics of the paCD-pDNA complexes (CDplexes) have been studied by using zeta potential, DLS, SAXS, and cryo-TEM. paCDs and pDNA, when assembled in CDplexes, render effective charges that are lower than the nominal ones. The CDplexes show a self-assembling pattern corresponding to multilamellar lyotropic liquid crystal phases, characterized by a lamellar stacking of bilayers of the CD-based vectors with anionic pDNA sandwiched among them. When exposed to human serum, either in the absence or in the presence of pDNA, the surface of the cationic CD-based vector becomes coated by a protein corona (PC) whose composition has been analyzed by nanoLC-MS/MS. Some of the CDplexes herein studied showed moderate-to-high transfection levels in HeLa and MCF-7 cancer cells combined with moderate-to-high cell viabilities, as determined by FACS and MTT reduction assays. The ensemble of data provides a detail picture of the paCD-pDNA-PC association processes and a rational base to exploit the protein corona for targeted gene delivery on future in vivo applications., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The synthesis of mono and divalent β-galactosylamides linked to a hydroxylated chain having a C2 symmetry axis derived from l-tartaric anhydride is reported. Reference compounds devoid of hydroxyl groups in the linker were also prepared from β-galactosylamine and succinic anhydride. After functionalization with an alkynyl residue, the resulting building blocks were grafted onto different azide-equipped scaffolds through the copper catalyzed azide-alkyne cycloaddition. Thus, a family of structurally related mono and divalent β-N-galactopyranosylamides was obtained and fully characterized. The binding affinities of the ligands towards the model lectin PNA were measured by the enzyme-linked lectin assay (ELLA). The IC 50 values were significantly higher than that of galactose but the presence of hydroxyl groups in the aglycone chain improved lectin recognition. Docking and molecular dynamics experiments were in accordance with the hypothesis that a hydroxyl group properly disposed in the linker could mimic the Glc O3 in the recognition process. On the other hand, divalent presentation of the ligands led to lectin affinity enhancements., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

This article shows the results of an experimental study carried out in order to determine the maximum amount of electric arc furnace dust (EAFD) that can be incorporated into fluid cement-based mortars to produce mechanically stable monolithic blocks. The leaching performance of all mixes was studied in order to classify them according to the EU Council Decision 2003/33/EC. Two mortars were used as reference and three levels of EAFD incorporation were tested in each of the reference mortars. As the incorporation ratio of EAFD/cement increases, the mechanical strength decreases. This is due to the greater EAFD/cement and water/cement ratios, besides the presence of a double-hydrated hydroxide of Ca and Zn (CaZn 2 (OH) 6 ·2H 2 O) instead of the portlandite phase (Ca(OH) 2 ) in the mixes made with EAFD, as well as non-hydrated tricalcium silicate. A mass ratio of 2:1 (EAFD: cement-based mortar) can be added maintaining a stable mechanical strength. The mechanical stabilization process also reduced the leaching of metals, although it was not able to reduce the Pb concentration below the limit for hazardous waste. The high amount of EAFD mechanically stabilized in this experimental study can be useful to reduce the storage volume required in hazardous waste landfills., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Polycationic carbohydrates represent an attractive class of biomolecules for several applications and particularly as non viral gene delivery vectors. In this case, the establishment of structure-biological activity relationship requires sensitive and accurate characterization tools to both control and achieve fine structural deciphering. Electrospray-tandem mass spectrometry (ESI-MS/MS) appears as a suitable approach to address these questions. In the study herein, we have investigated the usefulness of electron transfer dissociation (ETD) to get structural data about five polycationic carbohydrates demonstrated as promising gene delivery agents. A particular attention was paid to determine the influence of charge states as well as both fluoranthene reaction time and supplementary activation (SA) on production of charge reduced species, fragmentation yield, varying from 2 to 62%, as well as to obtain the most higher both diversity and intensity of fragments, according to charge states and targeted compounds. ETD fragmentation appeared to be mainly directed toward pending group rather than carbohydrate cyclic scaffold leading to a partial sequencing for building blocks when amino groups are close to carbohydrate core, but allowing to complete structural deciphering of some of them, such as those including dithioureidocysteaminyl group which was not possible with CID only. Such findings clearly highlight the potential to help the rational choice of the suitable analytical conditions, according to the nature of the gene delivery molecules exhibiting polycationic features. Moreover, our ETD-MS/MS approach open the way to a fine sequencing/identification of grafted groups carried on various sets of oligo-/polysaccharides in various fields such as glycobiology or nanomaterials, even with unknown or questionable extraction, synthesis or modification steps., (Copyright © 2016 Elsevier B.V. All rights reserved.)