Untargeted Metabolomic Analysis of Leaves and Roots of Jatropha curcas Genotypes with Contrasting Levels of Phorbol Esters.

AIMS: Phorbol esters (PE) are toxic diterpenoids accumulated in physic nut (Jatropha curcas L.) seed tissues. Their biosynthetic pathway remains unknown, and the participation of roots in this process may be possible. Thus, we set out to study the deposition pattern of PE and other terpenoids in roots and leaves of genotypes with detected (DPE) and not detected (NPE) phorbol esters based on previous studies. OUTLINE OF DATA RESOURCES: We analyzed physic nut leaf and root organic extracts using LC-HRMS. By an untargeted metabolomics approach, it was possible to annotate 496 and 146 metabolites in the positive and negative electrospray ionization modes, respectively. KEY RESULTS: PE were detected only in samples of the DPE genotype. Remarkably, PE were found in both leaves and roots, making this study the first report of PE in J. curcas roots. Furthermore, untargeted metabolomic analysis revealed that diterpenoids and apocarotenoids are preferentially accumulated in the DPE genotype in comparison with NPE, which may be linked to the divergence between the genotypes concerning PE biosynthesis, since sesquiterpenoids showed greater abundance in the NPE. UTILITY OF THE RESOURCE: The LC-HRMS files, publicly available in the MassIVE database (identifier MSV000092920), are valuable as they expand our understanding of PE biosynthesis, which can assist in the development of molecular strategies to reduce PE levels in toxic genotypes, making possible the food use of the seedcake, as well as its potential to contain high-quality spectral information about several other metabolites that may possess biological activity.

Emotional Exhaustion Scale (ECE): Psychometric Properties in a Sample of Portuguese University Students.

Academic emotional exhaustion is the first stage of academic burnout syndrome, and it is necessary to assess it and intervene early, as the consequences can lead to harmful effects on psychological well-being. The main objectives of this study were to explore the psychometric properties of the Emotional Exhaustion Scale (ECE); to assess the association with suffering and positive well-being, using the MHI-5 Inventory; and to provide information on its validity and to compare it with these constructs. A total sample of 526 university students (81% female and 19% male) aged between 17 and 62 (M = 21.42 years and SD = 5.78) took part in this study. Exploratory and confirmatory factor analyses were carried out. In the internal consistency assessment, the ECE was 0.89 and the MHI-5 was 0.81 (Cronbach's alpha). Exploratory factor analysis was carried out with Varimax rotation and confirmatory analysis, obtaining the factor that explains 50.5% of the variance. The results indicated that the ECE has adequate psychometric properties for use with higher education students in Portugal. Its use by mental health professionals in higher education institutions could be very relevant for screening for emotional exhaustion and thus preventing possible serious pathologies.

Modulation of Abundance and Location of High-Mobility Group Box 1 in Human Microglia and Macrophages under Oxygen-Glucose Deprivation.

While stroke represents one of the main causes of death worldwide, available effective drug treatment options remain limited to classic thrombolysis with recombinant tissue plasminogen activator (rtPA) for arterial-clot occlusion. Following stroke, multiple pathways become engaged in producing a vicious proinflammatory cycle through the release of damage-associated molecular patterns (DAMPs) such as high-mobility group box 1 (HMGB1) and heat shock protein 70 kDa (HSP72). HMGB1, in particular, can activate proinflammatory cytokine production when acetylated (AcHMGB1), a form that prefers cytosolic localization and extracellular release. This study aimed at determining how HMGB1 and HSP72 are modulated and affected following treatment with the anti-inflammatory compound resveratrol and novel platelet membrane-derived nanocarriers loaded with rtPA (CSM@rtPA) recently developed by our group for ischemic artery recanalization. Under ischemic conditions of oxygen-glucose deprivation (OGD), nuclear abundance of HMGB1 and AcHMGB1 in microglia and macrophages decreased, whereas treatment with CSM@rtPA did not alter nuclear or cytosolic abundance. Resveratrol treatment markedly increased the cytosolic abundance of HSP72 in microglia. Using proximity ligation assays, we determined that HSP72 interacted with HMGB1 and with acetylated HMGB1. The interaction was differentially affected under the OGD conditions. Resveratrol treatment under the OGD further decreased HSP72-HMGB1 interactions, whereas, in contrast, treatment increased HSP72-AcHMGB1 interactions in microglia. This study points out a salient molecular interaction suited for a two-pronged nanotherapeutic intervention in stroke: enhancement of rtPA's thrombolytic activity and modulation of cytosolic interactions between HMGB1 and HSP72 by resveratrol.

Pharmacological preclinical comparison of tenecteplase and alteplase for the treatment of acute stroke.

Alteplase (rtPA) remains the standard thrombolytic drug for acute ischemic stroke. However, new rtPA-derived molecules, such as tenecteplase (TNK), with prolonged half-lives following a single bolus administration, have been developed. Although TNK is currently under clinical evaluation, the limited preclinical data highlight the need for additional studies to elucidate its benefits. The toxicities of rtPA and TNK were evaluated in endothelial cells, astrocytes, and neuronal cells. In addition, their in vivo efficacy was independently assessed at two research centers using an ischemic thromboembolic mouse model. Both therapies were tested via early (20 and 30 min) and late administration (4 and 4.5 h) after stroke. rtPA, but not TNK, caused cell death only in neuronal cultures. Mice were less sensitive to thrombolytic therapies than humans, requiring doses 10-fold higher than the established clinical dose. A single bolus dose of 2.5 mg/kg TNK led to an infarct reduction similar to perfusion with 10 mg/kg of rtPA. Early administration of TNK decreased the hemorrhagic transformations compared to that by the early administration of rtPA; however, this result was not obtained following late administration. These two independent preclinical studies support the use of TNK as a promising reperfusion alternative to rtPA.

Systemic biomarker associated with poor outcome after futile reperfusion.

BACKGROUND: Successful recanalization does not lead to complete tissue reperfusion in a considerable percentage of ischemic stroke patients. This study aimed to identify biomarkers associated with futile recanalization. Leukoaraiosis predicts poor outcomes of this phenomenon. Soluble tumour necrosis factor-like weak inducer of apoptosis (sTWEAK), which is associated with leukoaraiosis degrees, could be a potential biomarker. METHODS: This study includes two cohorts of ischemic stroke patients in a multicentre retrospective observational study. Effective reperfusion, defined as a reduction of ≥8 points in the National Institutes of Health Stroke Scale (NIHSS) within the first 24 h, was used as a clinical marker of effective reperfusion. RESULTS: In the first cohort study, female sex, age, and high NIHSS at admission (44.7% vs. 81.1%, 71.3 ± 13.7 vs. 81.1 ± 6.7; 16 [13, 21] vs. 23 [17, 28] respectively; p < .0001) were confirmed as predictors of futile recanalization. ROC curve analysis showed that leukocyte levels (sensitivity of 99%, specificity of 55%) and sTWEAK level (sensitivity of 92%, specificity of 88%) can discriminate between poor and good outcomes. Both biomarkers simultaneously are higher associated with outcome after effective reperfusion (OR: 2.17; CI 95% 1.63-4.19; p < .0001) than individually (leukocytes OR: 1.38; CI 95% 1.00-1.64, p = .042; sTWEAK OR: 1.00; C I95% 1.00-1.01, p = .019). These results were validated using a second cohort, where leukocytes and sTWEAK showed a sensitivity of 100% and specificity of 66.7% and 75% respectively. CONCLUSIONS: Leukocyte and sTWEAK could be biomarkers of reperfusion failure and subsequent poor outcomes. Further studies will be necessary to explore its role in reperfusion processes.

Alternative conformations of a group 4 Late Embryogenesis Abundant protein associated to its in vitro protective activity.

Late Embryogenesis Abundant (LEA) proteins are a group of intrinsically disordered proteins implicated in plant responses to water deficit. In vitro studies revealed that LEA proteins protect reporter enzymes from inactivation during low water availability. Group 4 LEA proteins constitute a conserved protein family, displaying in vitro protective capabilities. Under water deficiency or macromolecular crowding, the N-terminal of these proteins adopts an alpha-helix conformation. This region has been identified as responsible for the protein in vitro protective activity. This study investigates whether the attainment of alpha-helix conformation and/or particular amino acid residues are required for the in vitro protective activity. The LEA4-5 protein from Arabidopsis thaliana was used to generate mutant proteins. The mutations altered conserved residues, deleted specific conserved regions, or introduced prolines to hinder alpha-helix formation. The results indicate that conserved residues are not essential for LEA4-5 protective function. Interestingly, the C-terminal region was found to contribute to this function. Moreover, alpha-helix conformation is necessary for the protective activity only when the C-terminal region is deleted. Overall, LEA4-5 shows the ability to adopt alternative functional conformations under the tested conditions. These findings shed light on the in vitro mechanisms by which LEA proteins protect against water deficit stress.

New Perspectives in Neuroprotection for Ischemic Stroke.

The constant failure of new neuroprotective therapies for ischemic stroke has partially halted the search for new therapies in recent years, mainly because of the high investment risk required to develop a new treatment for a complex pathology, such as stroke, with a narrow intervention window and associated comorbidities. However, owing to recent progress in understanding the stroke pathophysiology, improvement in patient care in stroke units, development of new imaging techniques, search for new biomarkers for early diagnosis, and increasingly widespread use of mechanical recanalization therapies, new opportunities have opened for the study of neuroprotection. This review summarizes the main protective agents currently in use, some of which are already in the clinical evaluation phase. It also includes an analysis of how recanalization therapies, new imaging techniques, and biomarkers have improved their efficacy.

Thrombolytic therapy based on lyophilized platelet-derived nanocarriers for ischemic stroke.

BACKGROUND: Intravenous administration of fibrinolytic drugs, such as recombinant tissue plasminogen activator (rtPA) is the standard treatment of acute thrombotic diseases. However, current fibrinolytics exhibit limited clinical efficacy because of their short plasma half-lives and risk of hemorrhagic transformations. Platelet membrane-based nanocarriers have received increasing attention for ischemic stroke therapies, as they have natural thrombus-targeting activity, can prolong half-life of the fibrinolytic therapy, and reduce side effects. In this study we have gone further in developing platelet-derived nanocarriers (defined as cellsomes) to encapsulate and protect rtPA from degradation. Following lyophilization and characterization, their formulation properties, biocompatibility, therapeutic effect, and risk of hemorrhages were later investigated in a thromboembolic model of stroke in mice. RESULTS: Cellsomes of 200 nm size and loaded with rtPA were generated from membrane fragments of human platelets. The lyophilization process did not influence the nanocarrier size distribution, morphology, and colloidal stability conferring particle preservation and long-term storage. Encapsulated rtPA in cellsomes and administered as a single bolus showed to be as effective as a continuous clinical perfusion of free rtPA at equal concentration, without increasing the risk of hemorrhagic transformations or provoking an inflammatory response. CONCLUSIONS: This study provides evidence for the safe and effective use of lyophilized biomimetic platelet-derived nanomedicine for precise thrombolytic treatment of acute ischemic stroke. In addition, this new nanoformulation could simplify the clinical use of rtPA as a single bolus, being easier and less time-consuming in an emergency setting than a treatment perfusion, particularly in stroke patients. We have successfully addressed one of the main barriers to drug application and commercialization, the long-term storage of nanomedicines, overcoming the potential chemical and physical instabilities of nanomedicines when stored in an aqueous buffer.

Comparison of microvascular decompression, percutaneous radiofrequency rhizotomy, and stereotactic radiosurgery in the treatment of trigeminal neuralgia: A long term quasi-experimental study.

BACKGROUND: Microvascular decompression (MVD), radiofrequency rhizotomy (RFR), and stereotactic radiosurgery (SRS) are surgical techniques frequently used in the treatment of idiopathic trigeminal neuralgia (TN), although the results reported for each of these are diverse. OBJECTIVE: This study aimed to compare long-term pain control obtained by MVD, SRS, and RFR in patients with idiopathic TN. METHODS: To compare the results obtained by MVD, SRS, and RFR we chose a quasi-experimental, ambispective design with control groups but no pretest. A total of 52 participants (MVD n = 33, RFR n = 10, SRS n = 9) were included. Using standardized outcome measures, pain intensity, pain relief, quality of life, and satisfaction with treatment were assessed by an independent investigator. The TREND statement for reporting non-randomized evaluations was applied. Clinical outcomes were evaluated at the initial postoperative period and at 6 months, 1, 2, 3, 4, and 5 years postoperatively. RESULTS: MVD has shown better results in pain scales compared to ablative procedures. Significant differences between groups were found regarding pain intensity and pain relief at the initial postoperative period (p < 0.001) and 6 months (p = 0.022), 1 year (p < 0.001), 2 years (p = 0.002), and 3 years (p = 0.004) after the intervention. Those differences exceeded the thresholds of the minimal clinically important difference. A higher percentage of patients free of pain was observed in the group of patients treated by MVD, with significant differences at the initial postoperative period (p < 0.001) and 6 months (p = 0.02), 1 year (p = 0.001), and 2 years (p = 0.04) after the procedure. Also, a higher risk of pain recurrence was observed in the RFR and SRS groups (HR 3.15, 95% CI 1.33-7.46; p = 0.009; and HR 4.26, 95% CI 1.77-10.2; p = 0.001, respectively) compared to the MVD group. No significant differences were found in terms of quality of life and satisfaction with treatment. A higher incidence of complications was observed in the MVD group. CONCLUSION: Concerning pain control and risk of pain recurrence, MVD is superior to RFR and SRS, but not in terms of quality of life, satisfaction with treatment, and safety profile.

Streptococcus pneumoniae serotype 19A in hospitalized children with invasive pneumococcal disease after the introduction of conjugated vaccines in Lima, Peru.

BACKGROUND: The Pneumococcal conjugate vaccine (PCV) has decreased cases of invasive pneumococcal disease (IPD) worldwide. However, the impact of PCVs introduction may be affected by the serotype distribution in a specific context. METHODS: Cross-sectional multicenter passive surveillance study of IPD cases in pediatric patients hospitalized in Lima, Peru between 2016 and 2019 (after PCV13 introduction) to determine the serotype distribution and antimicrobial resistance of Streptococcus pneumoniae. Serotyping was performed by a sequential multiplex PCR and confirmed by whole genome sequencing. RESULTS: Eighty-five S. pneumoniae isolates were recovered (4.07/100,000 among children <60 months of age). Serotype 19A was the most common (49.4%). Children infected with serotype 19A in comparison with children infected with other serotypes were younger, had a lower rate of meningitis and higher rates of pneumonia, complicated pneumonia and antimicrobial resistance; 28.6% of patients with serotype 19A have received at least one dose of PCV13 vs. 62.8% of patients with other serotypes. Using MIC-breakpoints, 81.2% (56/69) of non-meningitis strains and 31.2% (5/16) of meningitis strains were susceptible to penicillin; 18.8% (3/16) of meningitis strains had intermediate resistance to ceftriaxone. Resistance to azithromycin was 78.8% (67/85). Serotype 19A frequency increased over time in the same study population, from 4.2% (4/96) in 2006-2008, to 8.6% (5/58) in 2009-2011, to 49.4% (42/85) in the current study (2016-2019) (p < 0.001). CONCLUSIONS: After PCV13 introduction in Peru, serotype 19A remains the most prevalent; however, the vaccination coverage is still not optimal. Therefore, additonal surveillance studies are needed to determine the remaining IPD burden.

Impact of Exercise Interventions on Sustained Brain Health Outcomes in Frail Older Individuals: A Comprehensive Review of Systematic Reviews.

Several systematic review studies highlight exercise's positive impact on brain health outcomes for frail individuals. This study adopts a Comprehensive Review of reviews (CRs) approach to amalgamate data from existing reviews, focusing on exercise's influence on brain health outcomes in older frail and pre-frail adults. The methodology involves a thorough search of Portuguese, Spanish, and English-indexed databases (i.e., Ebsco Health, Scielo, ERIC, LILACS, Medline, Web of Science, SportDiscus) from 1990 to 2022, with the AMSTAR-2 tool assessing evidence robustness. The search terms "physical exercise", "elderly frail", and "systematic review" were employed. Results: Out of 12 systematically reviewed studies, four presented high-quality (with metanalyses), while eight exhibit critically low quality. Positive trends emerge in specific cognitive and neuromotor aspects, yet challenges persist in psychosocial domains, complex cognitive tasks, and ADL outcomes. This study yields reasonable and promising evidence regarding exercise's influence on quality of life and depression in frail older individuals. However, the impact on biochemical markers remains inconclusive, emphasizing the need for standardized methodologies. Conclusions: The findings highlight the importance of acknowledging methodological nuances for clinicians and policymakers when translating these results into impactful interventions for aging populations. This emphasizes the necessity for a comprehensive and customized approach to exercise interventions aimed at fostering the sustainability of overall well-being in older individuals, aligning with United Nations Sustainable Development Goal 3.

Active Neurodynamics at Home in Patients with Knee Osteoarthritis: A Feasibility Study.

The aim was to evaluate the feasibility of a home-based neurodynamic programme for patients with knee osteoarthritis (KO). Thirty participants (70% women) ≥ 50 years old with KO (Kellgren-Lawrence grades I-II) were included. Active mobilisation of the femoral nerve was performed at home over a period of 6-8 weeks. The feasibility of the programme was assessed using a survey that included questions related to understanding of the activity; adherence to the intervention; the burden caused by the intervention; self-perceived effects on the participant; follow-up; the barriers; and facilitators. Pain intensity, using the numerical rating scale (NRS); pressure pain thresholds (PPT); temporal assessment; pain modulation; Knee Injury and Osteoarthritis Outcome Score (KOOS), 12-item Short Form Survey questionnaire (SF-12), and the Central Sensitization Inventory questionnaire (CSI) were also collected, before and after the intervention. All patients performed the intervention, completed at least 42 days of activity, and considered the exercise adequate, with 28 participants (93.3%) reporting that the intervention was good for them. Statistically significant values (p < 0.05) were found for NRS, elbow PPT, external knee PPT, internal knee PPT, elbow CPM, CSI, and KOOS. Home-based active neurodynamic treatment has been shown to be a feasible and safe intervention for KO patients. In addition, this intervention has shown positive effects on pain and function.

Minimally Invasive Retrosigmoidal Parasterional Burr-Hole Approach: Technique and Neuropathic Pain Amelioration after Microvascular Decompression of the Trigeminal Nerve.

BACKGROUND: Trigeminal neuralgia, a common condition in clinical practice, often occurs due to vascular compression caused by aberrant or ectopic arterial or venous vessels. Microvascular decompression through a minimally invasive retrosigmoidal approach has shown high rates of pain control, low complication rates, and excellent therapeutic results. OBJECTIVE: To describe the surgical technique and clinical outcomes in terms of pain relief after microvascular decompression of the trigeminal nerve through a minimally invasive retrosigmoidal parasterional burr-hole technique. METHODS: A group of patients with trigeminal neuralgia refractory to medical management who underwent microvascular decompression were examined. The records of the patients were considered retrospectively (2016-2018), and the outcomes were considered based on the Visual Analogue Scale (VAS) and the Barrow Neurological Institute Pain Scale (BNIPS) added to a technical note of the surgical technique for a minimally invasive retrosigmoidal parasterional burr-hole. RESULTS: Twenty-two patients were evaluated, and clinical assessment after surgical intervention showed a decrease in pain according to the VAS, resulting from an average preoperative state of 9.5 ± 0.37 to a postoperative condition of 1.32 ± 1.28, exhibiting statistically significant changes (p < 0.0001, d = 9.356). On the other hand, in relation to the BNIPS scale, a decrease from an average preoperative status of 4.55 ± 0.25 to a postoperative status at 12 months of 1.73 ± 0.54 was also demonstrated, showing significant changes (p < 0.0001, d = 3.960). CONCLUSION: Microvascular decompression of the trigeminal nerve through a minimally invasive retrosigmoidal parasterional burr-hole is feasible and can be a safe and effective technique for the management of pain. However, further research employing larger sample sizes and longer follow-up periods is necessary.

Transformations in the world of healthcare work: workers and future challenges. / Transformações no mundo do trabalho em saúde: os(as) trabalhadores(as) e desafios futuros.

This article addresses the world of healthcare work, especially in the Brazilian Unified Healthcare System (SUS) in the context of the COVID-19 pandemic in Brazil. This study used data from the following surveys: "Working conditions of health professionals in the context of COVID-19 in Brazil" and "The Invisible health workers: working conditions and mental health in the context of COVID-19 in Brazil". Data analysis proves that the pandemic highlighted existing structural problems within SUS, involving the issue of healthcare workforce (HWF) management, which can be interpreted as another reflection of the socioeconomic inequalities that already exist in the country. This article highlights: the reduced provision of permanent education, the regulation of hybrid care, precariousness, a lack of protection in the work environment, as well as fragile biosecurity leading to tragic rates of illness and death of health workers. Our study concludes by showing the importance of formulating public policies in the scope of education and work management in SUS that ensure the discussion on hybrid care as a new way of acting without losing quality, together with the need to review issues related to permanent education, protection, valuation, and reduction of inequalities pointed out among the professional contingents analyzed in this article.

O artigo versa sobre o mundo do trabalho da saúde, especialmente no SUS no contexto da pandemia no Brasil. O artigo utilizou dados das pesquisas "Condições de trabalho dos profissionais de saúde no contexto da COVID-19 no Brasil" e "Os trabalhadores invisíveis da saúde: condições de trabalho e saúde mental no contexto da COVID-19 no Brasil". A análise dos dados comprova que a pandemia evidenciou problemas estruturais existentes no âmbito do Sistema Único de Saúde, envolvendo a gestão da FTS o que pode ser interpretado como mais um dos reflexos das desigualdades socioeconômicas já existentes no país. Destacam-se: a reduzida oferta de educação permanente, a regulação do cuidado híbrido, precarização, desproteção no ambiente de trabalho, frágil biossegurança levando a trágicas taxas de adoecimento e mortes de trabalhadores da saúde. Conclui mostrando a importância de formulação de políticas públicas no âmbito da gestão da educação e do trabalho no SUS que assegurem a discussão sobre cuidado híbrido como nova forma de atuar sem perder qualidade, a necessidade de se rever questões referentes a: educação permanente, proteção, valorização e redução das desigualdades apontadas entre os contingentes profissionais analisados nesse artigo.

Transformações no mundo do trabalho em saúde: os(as) trabalhadores(as) e desafios futuros / Transformations in the world of healthcare work: workers and future challenges

Resumo O artigo versa sobre o mundo do trabalho da saúde, especialmente no SUS no contexto da pandemia no Brasil. O artigo utilizou dados das pesquisas "Condições de trabalho dos profissionais de saúde no contexto da COVID-19 no Brasil" e "Os trabalhadores invisíveis da saúde: condições de trabalho e saúde mental no contexto da COVID-19 no Brasil". A análise dos dados comprova que a pandemia evidenciou problemas estruturais existentes no âmbito do Sistema Único de Saúde, envolvendo a gestão da FTS o que pode ser interpretado como mais um dos reflexos das desigualdades socioeconômicas já existentes no país. Destacam-se: a reduzida oferta de educação permanente, a regulação do cuidado híbrido, precarização, desproteção no ambiente de trabalho, frágil biossegurança levando a trágicas taxas de adoecimento e mortes de trabalhadores da saúde. Conclui mostrando a importância de formulação de políticas públicas no âmbito da gestão da educação e do trabalho no SUS que assegurem a discussão sobre cuidado híbrido como nova forma de atuar sem perder qualidade, a necessidade de se rever questões referentes a: educação permanente, proteção, valorização e redução das desigualdades apontadas entre os contingentes profissionais analisados nesse artigo.

Abstract This article addresses the world of healthcare work, especially in the Brazilian Unified Healthcare System (SUS) in the context of the COVID-19 pandemic in Brazil. This study used data from the following surveys: "Working conditions of health professionals in the context of COVID-19 in Brazil" and "The Invisible health workers: working conditions and mental health in the context of COVID-19 in Brazil". Data analysis proves that the pandemic highlighted existing structural problems within SUS, involving the issue of healthcare workforce (HWF) management, which can be interpreted as another reflection of the socioeconomic inequalities that already exist in the country. This article highlights: the reduced provision of permanent education, the regulation of hybrid care, precariousness, a lack of protection in the work environment, as well as fragile biosecurity leading to tragic rates of illness and death of health workers. Our study concludes by showing the importance of formulating public policies in the scope of education and work management in SUS that ensure the discussion on hybrid care as a new way of acting without losing quality, together with the need to review issues related to permanent education, protection, valuation, and reduction of inequalities pointed out among the professional contingents analyzed in this article.

Magnetic separation and concentration of Aß 1-42 molecules dispersed at the threshold concentration for Alzheimer's disease diagnosis in clinically-relevant volumes of sample.

BACKGROUND: Alzheimer's disease (AD) is the leading cause of dementia and loss of autonomy in the elderly, implying a progressive cognitive decline and limitation of social activities. The progressive aging of the population is expected to exacerbate this problem in the next decades. Therefore, there is an urgent need to develop quantitative diagnostic methodologies to assess the onset the disease and its progression especially in the initial phases. RESULTS: Here we describe a novel technology to extract one of the most important molecular biomarkers of AD (Aß1-42) from a clinically-relevant volume - 100 µl - therein dispersed in a range of concentrations critical for AD early diagnosis. We demonstrate that it is possible to immunocapture Aß1-42 on 20 nm wide magnetic nanoparticles functionalized with hyperbranced KVLFF aptamers. Then, it is possible to transport them through microfluidic environments to a detection system where virtually all (~ 90%) the Aß1-42 molecules are concentrated in a dense plug of ca.50 nl. The technology is based on magnetic actuation by permanent magnets, specifically designed to generate high gradient magnetic fields. These fields, applied through submillimeter-wide channels, can concentrate, and confine magnetic nanoparticles (MNPs) into a droplet with an optimized shape that maximizes the probability of capturing highly diluted molecular biomarkers. These advancements are expected to provide efficient protocols for the concentration and manipulation of molecular biomarkers from clinical samples, enhancing the accuracy and the sensitivity of diagnostic technologies. CONCLUSIONS: This easy to automate technology allows an efficient separation of AD molecular biomarkers from volumes of biological solutions complying with the current clinical protocols and, ultimately, leads to accurate measurements of biomarkers. The technology paves a new way for a quantitative AD diagnosis at the earliest stage and it is also adaptable for the biomarker analysis of other pathologies.

Efficacy of an oral collagen therapy compared with intra-articular therapies (hyaluronic acid and platelet-rich plasma) to treat knee osteoarthritis / Eficacia de una terapia oral de colágeno comparada con terapias intraarticulares (ácido hialurónico y plasma rico en plaquetas) en el manejo de la osteoartritis de rodilla

Introducción: la osteoartritis es una enfermedad crónica y progresiva que afecta a más del 30 % de las personas mayores de 60 años. Actualmente, se reconoce la osteoartritis como una enfermedad multifactorial. Se emplean varios tratamientos conservadores en el manejo de la osteoartritis de rodilla (AINE, analgésicos y terapias intraarticulares). Se realizó un ensayo clínico aleatorizado para determinar si una terapia a base de 10 g de colágeno hidrolizado y 100 mg de fucoidano (Hydroidan pro, Acten, Suiza) era más efectiva que las terapias intraarticulares. Métodos: se dividió a los pacientes en 3 grupos. El primero recibió 23 g de ACTEN® cada día durante 3 meses. Los otros grupos recibieron una única inyección intraarticular de ácido hialurónico (5 ml) o plasma rico en plaquetas (3 ml). Se emplearon las escalas WOMAC, SF-12 y VAS para valorar el dolor al inicio, 4, 12 y 24 semanas después. Resultados: se incluyó a 108 pacientes con osteoartritis de rodilla de grado II-III que participaron en un estudio de seguimiento de 24 semanas de duración. La edad media fue de 57 años (53-65). Los 3 grupos rebajaron la puntuación en el grupo WOMAC (p < 0,001). El grupo que recibió colágeno y fucoidano obtuvo puntuaciones más bajas en las escalas WOMAC y VAS que los grupos que recibieron ácido hialurónico y plasma rico en plaquetas a las 24 semanas (p < 0,001). Conclusiones: el colágeno y el fucoidano tomado por vía oral, a diario, y durante 12 semanas parecen cosechar mejores resultados en las escalas WOMAC y VAS que las terapias intraarticulares a base de ácido hialurónico o plasma rico en plaquetas. Se debería de intentarse combinar terapias orales e intraarticulares para determinar su perfil de eficacia.(AU)

Introduction: osteoarthritis is a chronic and progressive disease. It affects over 30 % of people older than 60. Osteoarthri-tis is currently recognized as a multifactorial disease. Various conservative treatments are used in the management of kneeosteoarthritis (NSAIDs, analgesics, and intra-articular therapy). We conducted a randomized clinical trial to determine if a10 g therapy of hydrolyzed collagen along with 100 mg fucoidan (Hydroidan pro, Acten, Switzerland) is more effective thanintra-articular therapies.Methods: we divided patients into 3 groups. The first group received 23 g of ACTEN®, daily, for 3 months. The other groupsreceived a single intra-articular injection of hyaluronic acid (5 ml) or platelet-rich plasma (3 ml). We used the WOMAC scale,the SF-12 scale, and the VAS for pain at baseline, and 4, 12, and 24 weeks later.Results: we enrolled 108 patients with grade II-III knee osteoarthritis who underwent a 24-week follow-up study. The meanage was 57 years (53-65). The three groups showed low scores in the WOMAC group (p < 0.001). The collagen with fucoid-an group had lower WOMAC and VAS scores compared with the hyaluronic acid and platelet-rich plasma groups at 24weeks (p < 0.001).Conclusions: collagen along with fucoidan taken orally, daily, for 12 weeks seem to have better results in the WOMACand VAS scales compared with intra-articular therapies such as hyaluronic acid or platelet-rich plasma. Combined oral andintra-articular therapies should be tried to determine their efficacy profile.(AU)

The Gaze Patterns of Group Fitness Instructors Based on Different Levels of Training and Professional Experience.

The way in which group fitness instructors observe participants has a great influence on their pedagogical intervention. Based on the above, the main objective of this research is to characterize and compare their gaze patterns according to their training and professional experience. Twenty group fitness instructors of choreographed classes participated, aged between 18 and 42 years old, and for the comparison, four groups were created. Eye movements were captured with TOBII Pro Glasses 3, and data were coded and analyzed using the TOBII Pro Lab software. For the characterization of the gaze patterns, descriptive statistics were used in terms of count (f/m) and duration (s/m), while the comparison was performed using a one-way ANOVA test. More trained and experienced instructors tend to look at participants less, in count (80.59 ± 0.74) and duration (17.74 ± 0.71), with significant differences between the groups in some areas of interest (head, lower body, and other). There are also significant differences in the total number of eye fixations (F = 34.614; p = 0.001; η2 = 0.866; effect size very high). In conclusion, and projecting future works, it is important to understand how these gaze patterns are related to pedagogical behaviors in general or based on some specific factors (e.g., pedagogical feedback).

Analysis of antimicrobial efficacy of sodium hypochlorite and ozonated water against biofilm in oval canals.

This in vitro study compared the antimicrobial efficacy of 2.5% sodium hypochlorite (NaOCl) and 8 µg/mL ozonated water agitated by passive ultrasonic irrigation (PUI) or PUI combined with EndoActivator (EA) against mature multispecies biofilm. One hundred and five oval-shaped mandibular premolars were instrumented, sterilized, and inoculated with Enterococcus faecalis, Candida albicans, and Staphylococcus aureus, divided into: control group - saline; O3 group - ozonated water; O3 PUI group - ozonated water with PUI agitation; O3 PUI+EA group - ozonated water with PUI+EA agitation; NaOCl group - NaOCl; NaOCl PUI group - NaOCl with PUI agitation; and NaOCl PUI+EA group - NaOCl with PUI+EA agitation. Microbiological samples were collected before (S1) and after (S2) the disinfection procedures and the data were statistically analyzed using the Kruskal-Wallis test. In the culture method, there was significant disinfection in the O3 PUI+EA, NaOCl, NaOCl PUI, and NaOCl PUI+EA groups (p˂0.05). The combination of NaOCl with PUI+EA reduced microbial counts to zero (p˂0.05). In the qPCR method, there was a significant reduction in the total count of viable microorganisms in the O3 PUI, O3 PUI+EA, NaOCl, NaOCl PUI, and NaOCl PUI+EA groups (p˂0.05). It can be concluded that 2.5% NaOCl with and without agitation, as well as 8 µg/mL ozonated water with its action enhanced by the agitation techniques, were effective in root canal disinfection, and their antimicrobial efficacy is related to the microorganisms present in the biofilm.