Genomic insights into a Proteus mirabilis strain inducing avian cellulitis.

Proteus mirabilis, a microorganism distributed in soil, water, and animals, is clinically known for causing urinary tract infections in humans. However, recent studies have linked it to skin infections in broiler chickens, termed avian cellulitis, which poses a threat to animal welfare. While Avian Pathogenic Escherichia coli (APEC) is the primary cause of avian cellulitis, few cases of P. mirabilis involvement are reported, raising questions about the factors facilitating such occurrences. This study employed a pan-genomic approach to investigate whether unique genes exist in P. mirabilis strains causing avian cellulitis. The genome of LBUEL-A33, a P. mirabilis strain known to cause this infection, was assembled, and compared with other P. mirabilis strains isolated from poultry and other sources. Additionally, in silico serogroup analysis was conducted. Results revealed numerous genes unique to the LBUEL-A33 strain. No function in cellulitis was identified for these genes, and in silico investigation of the virulence potential of LBUEL-A33's exclusive proteins proved inconclusive. These findings support that multiple factors are necessary for P. mirabilis to cause avian cellulitis. Furthermore, this species likely employs its own unique arsenal of virulence factors, as many identified mechanisms are analogous to those of E. coli. While antigenic gene clusters responsible for serogroups were identified, no clear trend was observed, and the gene cluster of LBUEL-A33 did not show homology with any sequenced Proteus serogroups. These results reinforce the understanding that this disease is multifactorial, necessitating further research to unravel the mechanisms and underpin the development of control and prevention strategies.

Social vulnerability among Brazilian children in early childhood: a scoping review.

OBJECTIVE: To identify, map, and describe studies involving Brazilian children in early childhood in situations of social vulnerability. SOURCE OF DATA: A scoping review including full articles published in Portuguese and English up to March 2023, with no temporal restrictions. Searches were conducted in the MEDLINE/PubMed, Scielo, EMBASE, Cochrane, Scopus, CINAHL, Web of Science, PEDro, and LILACS databases. Journal metrics, sample characteristics, study area, characterization of the situation of social vulnerability, and study outcomes were extracted. SUMMARY OF THE FINDINGS: Seventy-six articles involving a total of 107.740 children in early childhood were included in this study. These studies presented relevant findings, including the temporal publication trend, the variability of social vulnerability indicators, the scarcity of intervention studies, and the fact that 100% of eligible studies were from the health area. Negative outcomes were associated with the condition of social vulnerability in almost the entire sample, reinforcing the need for government policies capable of protecting early childhood from the effects of social vulnerability. CONCLUSION: This scoping review mapped important findings involving Brazilian children in social vulnerability. It also identified literature gaps such as the need for intervention and multisectoral studies among health, education, and social protection.

Exploring the Relationship Between Genetic Instability and Health Outcomes in Acute and Chronic Post-COVID Syndrome.

The COVID-19 pandemic has led to the emergence of acute and chronic post-COVID syndromes, which present diverse clinical manifestations. The underlying pathophysiology of these conditions is not yet fully understood, but genetic instability has been proposed as a potential contributing factor. This study aimed to explore the differential impact of physical and psychological health factors on genetic instability in individuals with acute and chronic post-COVID syndromes. In this study, three groups of subjects were analyzed: a control group, an acute post-COVID group, and a chronic post-COVID group, with a total of 231 participants. The participants were assessed using a questionnaire for long-COVID-19COVID, and female participants reported more symptoms than male participants in areas related to fatigue, memory, mental health, and well-being during the chronic phase. Genetic instability was assessed using the comet assay, and participants' physical and psychological profiles were evaluated. The overall results showed no significant differences in DNA damage, as measured by the comet assay, among the three groups, suggesting that genetic instability, as assessed by this method, may not be a primary driver of the distinct clinical presentations observed in post-COVID syndromes. However, when gender was considered, male participants in the acute long COVID group exhibited higher levels of genetic instability compared to females. Multiple linear regression analysis revealed that gender, age, and waist circumference were significant predictors of DNA damage. Among females in the acute group, sexual health, and eye-related symptoms significantly influenced the increase in DNA damage. These findings indicate the need for further investigation on the gender-specific differences in genetic instability and their potential implications for the pathophysiology of post-COVID syndromes. Exploring alternative markers of genetic instability and the interplay between genetic, inflammatory, and cellular processes could provide valuable insights for the management of these debilitating post-viral sequelae.

Bixin, a New Atheroprotective Carotenoid Candidate, Prevents oxLDL-Induced Cytotoxicity and Mitochondrial Dysfunction in Macrophages: Involvement of the Nrf2 and NF-κB Pathways.

The accumulation of oxidized low-density lipoprotein (oxLDL) and its toxicity in the arterial wall have been implicated in atherosclerosis. This study aimed to investigate the mechanisms underlying the atheroprotective effect of bixin, a carotenoid obtained from the seeds of the tropical plant Bixa orellana, on Cu2+-induced LDL oxidation and oxLDL-mediated effects in J774A.1 macrophage cells. Bixin's effects were compared to those of lycopene, a carotenoid widely studied for its cardiovascular protective effects. LDL was isolated from human plasma, incubated with bixin or lycopene (positive control), and subjected to oxidation with CuSO4. Afterward, bixin or lycopene was incubated with J774A.1 macrophage cells and exposed to oxLDL. The levels of ROS, RNS, GSH, nitrite, mitochondrial function, and foam cell formation, as well as the expression of proteins related to the antioxidant and inflammatory status, were evaluated. The effect of bixin in inhibiting in vitro human-isolated LDL oxidation was more potent (5-6-fold) than that of lycopene. Bixin pretreatment reduced the atherogenic signaling triggered by oxLDL in the macrophages, namely the generation of reactive species, disturbance of nitric oxide homeostasis, mitochondrial dysfunction, and foam cell formation. The cytoprotective effects of bixin were accompanied by the upregulation of Nrf2 and the downregulation of the NF-kB pathways. Lycopene showed the same protective effect as bixin, except that it did not prevent mitochondrial dysfunction. The efficient performance of bixin makes it an ideal candidate for further trials as a new nutraceutical compound for the prevention of atherosclerosis.

Prediction of adverse drug reactions in geriatric patients admitted to intensive care units. / [Artículo traducido] Predicción de reacciones adversas a medicamentos en pacientes geriátricos ingresados en unidades de cuidados intensivos.

INTRODUCTION: Intensive Care Units (ICUs) pose challenges in managing critically-ill patients with polypharmacy, potentially leading to Adverse Drug Reactions (ADRs), particularly in the elderly. OBJECTIVE: To evaluate whether the severity and clinical prognosis scores used in ICUs correlate with the prediction of ADRs in aged patients admitted to an ICU. METHODS: A cohort study was conducted in a Brazilian University Hospital ICU. APACHE II and SAPS 3 assessed clinical prognosis, while GerontoNet ADR Risk Score and BADRI evaluated ADR risk at ICU admission. Severity of the patients' clinical conditions was evaluated daily based on the SOFA score. Adverse Drug Reaction (ADR) screening was performed daily through the identification of ADR triggers. RESULTS: 1295 triggers were identified (median 30 per patient, IQR = 28), with 15 suspected ADRs. No correlation was observed between patient severity and ADRs at admission (p=0.26), during hospitalization (p=0.91), or at follow-up (p=0.77). There was also no association between death and ADRs (p=0.28) or worse prognosis and ADRs (p>0.05). Higher BADRI scores correlated with more ADRs (p=0.001). CONCLUSIONS: The data suggest that employing the severity and clinical prognosis scores used in Intensive Care Units is not sufficient to direct active pharmacovigilance efforts, which are therefore indicated for critically ill patients.

Effect of VAChT reduction on lung alterations induced by exposure to iron particles in an asthma model.

INTRODUCTION: Pollution harms the health of people with asthma. The effect of the anti-inflammatory cholinergic pathway in chronic allergic inflammation associated to pollution is poorly understood. METHODS: One hundred eight animals were divided into 18 groups (6 animals). Groups included: wild type mice (WT), genetically modified with reduced VAChT (VAChTKD), and those sensitized with ovalbumin (VAChTKDA), exposed to metal powder due to iron pelletizing in mining company (Local1) or 3.21 miles away from a mining company (Local2) in their locations for 2 weeks during summer and winter seasons. It was analyzed for hyperresponsivity, inflammation, remodeling, oxidative stress responses and the cholinergic system. RESULTS: During summer, animals without changes in the cholinergic system revealed that Local1 exposure increased the hyperresponsiveness (%Rrs, %Raw), and inflammation (IL-17) relative to vivarium animals, while animals exposed to Local2 also exhibited elevated IL-17. During winter, animals without changes in the cholinergic system revealed that Local2 exposure increased the hyperresponsiveness (%Rrs) relative to vivarium animals. Comparing the exposure local of these animals during summer, animals exposed to Local1 showed elevated %Rrs, Raw, and IL-5 compared to Local 2, while in winter, Local2 exposure led to more IL-17 than Local1. Animals with VAChT attenuation displayed increased %Rrs, NFkappaB, IL-5, and IL-13 but reduced alpha-7 compared to animals without changes in the cholinergic system WT. Animals with VAChT attenuation and asthma showed increased the hyperresponsiveness, all inflammatory markers, remodeling and oxidative stress compared to animals without chronic lung inflammation. Exposure to Local1 exacerbated the hyperresponsiveness, oxidative stressand inflammation in animals with VAChT attenuation associated asthma, while Local2 exposure led to increased inflammation, remodeling and oxidative stress. CONCLUSIONS: Reduced cholinergic signaling amplifies lung inflammation in a model of chronic allergic lung inflammation. Furthermore, when associated with pollution, it can aggravate specific responses related to inflammation, oxidative stress, and remodeling.

Application of the participatory design in the testing of a baropodometric insole prototype for weight-bearing asymmetry after a stroke: A qualitative study.

Introduction: Currently studies indicate the need to incorporate the user`s perspective in the testing of new assistive technologies. The objective of this paper is to test a baropodometric insole prototype for monitoring and treatment weight-bearing asymmetry, according to the Participatory Design. Methods: We used a qualitative case study approach during the testing phase of the baropodometric insole prototype. The focus group approach addressed topics related to the experience and accessibility of the potential user in conjunction with professionals, researchers, and physiotherapy students. Facilitators, barriers, and requirements for the device were collected through audio recordings of the discussions during and after prototype testing. Results: Key steps in the prototype testing process were divided into (1) Test of the prototype according to the Participatory Design, divided into Who, When, How, and Why the potential user was involved in the study; and (2) Facilitators, barriers and requirements to improve the prototype. Conclusions: The baropodometric insole prototype can be seen as a promising device for monitoring and treating weight-bearing asymmetry.

Influence of Pressing Temperatures on Physical-Mechanical Properties of Wood Particleboards Made with Urea-Formaldehyde Adhesive Containing Al2O3 and CuO Nanoparticles.

Particleboards have gained attention in the global market. Understanding their physical-mechanical behavior in the current technological context is essential due to adhesive polymerization, which depends on variables such as pressing time and temperature. Today, the use of nanoparticles has become a plausible option for improving the properties of polymers used in wood-based composites. This study evaluates the influences of the addition of non-commercial 0.5% aluminum oxide (Al2O3) and aluminum oxide copper (CuO) nanoparticles using a greener route with a lower environmental impact obtaining a urea-formaldehyde (UF)-based polymeric adhesive to manufacture particle composites of Eucalyptus urophylla var. grandis wood. Regarding characterizations, the resin properties analyzed were viscosity, gel time, and pH, as well as panel properties, including density, moisture content, thickness swelling, modulus of elasticity, modulus of rupture, and thermal conductivity. The results were compared with scientific publications and standards. The addition of nanoparticles interfered with viscosity, and all treatments indicated a basic pH. It was not possible to determine the gel time after 10 min. Nanoparticles added to the polymers in the internal layer did not cause an improvement in the swelling properties in terms of thickness, with no significant statistical difference for density and moisture content. The increase from 150 °C to 180 °C may have caused an improvement in all physical-mechanical properties, indicating that the higher temperature positively influenced the polymerization of the formaldehyde-based adhesive. Therefore, the additions of both nanoparticles (0.5% in each condition) led to a limitation in the material influence with respect to physical-mechanical performance.

Effects of plant protease inhibitors (Pep-3-EcTI, Pep-BbKI, and Pep-BrTI) versus corticosteroids on inflammation, remodeling, and oxidative stress in an asthma-COPD (ACO) model.

The peptide derived from E. contortisiliquum trypsin inhibitor (Pep-3-EcTI), peptide derived from kallikrein inhibitor isolated from B. bauhinioides (Pep-BbKI), and B. rufa peptide modified from B. bauhinioides (Pep-BrTI) peptides exhibit anti-inflammatory and antioxidant activities, suggesting their potential for treating asthma-chronic obstructive pulmonary disease (COPD) overlap (ACO). We compared the effects of these peptides with dexamethasone (DX) treatment in an ACO model. In this study, 11 groups of male BALB/c mice were pre-treated under different conditions, including sensitization with intraperitoneal injection and inhalation of ovalbumin (OVA), intratracheal instillation of porcine pancreatic elastase (ELA), sensitization with intraperitoneal injection, and various combinations of peptide treatments with Pep-3-EcTI, Pep-BbKI, Pep-BrTI, dexamethasone, and non-treated controls (SAL-saline). Respiratory system resistance, airway resistance, lung tissue resistance, exhaled nitric oxide, linear mean intercept, immune cell counts in the bronchoalveolar lavage fluid, cytokine expression, extracellular matrix remodeling, and oxidative stress in the airways and alveolar septa were evaluated on day 28. Results showed increased respiratory parameters, inflammatory markers, and tissue remodeling in the ACO group compared to controls. Treatment with the peptides or DX attenuated or reversed these responses, with the peptides showing effectiveness in controlling hyperresponsiveness, inflammation, remodeling, and oxidative stress markers. These peptides demonstrated an efficacy comparable to that of corticosteroids in the ACO model. However, this study highlights the need for further research to assess their safety, mechanisms of action, and potential translation to clinical studies before considering these peptides for human use.

Functional Physical Analysis and Quality of Life in the Preoperative and Early Postoperative Periods of Cardiac Surgery and 30 Days After Hospital Discharge.

INTRODUCTION: The analysis of patients submitted to heart surgery at three assessment times has been insufficiently described in the literature. OBJECTIVE: To analyze chest expansion, maximum inspiratory pressure (MIP), maximum expiratory pressure (MEP), distance traveled on the six-minute walk test (6MWT), and quality of life in the preoperative period, fourth postoperative day (4th PO), and 30th day after hospital discharge (30th-day HD) in individuals submitted to elective heart surgery. METHODS: A descriptive, analytical, cross-sectional study was conducted with 15 individuals submitted to elective heart surgery between 2016 and 2020 who did not undergo any type of physiotherapeutic intervention in Phase II of cardiac rehabilitation. The outcome variables were difference in chest expansion (axillary, nipple, and xiphoid), MIP, MEP, distance on 6MWT, and quality of life. The assessment times were preoperative period, 4th PO, and 30th-day HD. RESULTS: Chest expansion diminished between the preoperative period and 4th PO, followed by an increase at 30th-day HD. MIP, MEP, and distance traveled on the 6MWT diminished between the preoperative period and 4th PO, with a return to preoperative values at 30th-day HD. General quality of life improved between the preoperative period and 4th PO and 30th-day HD. An improvement was found in the social domain between the preoperative period and the 30th-day HD. CONCLUSION: Heart surgery causes immediate physical deficit, but physical functioning can be recovered 30 days after hospital discharge, resulting in an improvement in quality of life one month after surgery.

Prediction of adverse drug reactions in geriatric patients admitted to intensive care units.

INTRODUCTION: Intensive care units (ICUs) pose challenges in managing critically ill patients with polypharmacy, potentially leading to adverse drug reactions (ADRs), particularly in the elderly. OBJECTIVE: To evaluate whether the severity and clinical prognosis scores used in ICUs correlate with the prediction of ADRs in aged patients admitted to an ICU. METHODS: A cohort study was conducted in a Brazilian University Hospital ICU. APACHE II and SAPS 3 assessed clinical prognosis, while GerontoNet ADR Risk Score and BADRI evaluated ADR risk at ICU admission. Severity of the patients' clinical conditions was evaluated daily based on the SOFA score. ADR screening was performed daily through the identification of ADR triggers. RESULTS: 1295 triggers were identified (median 30 per patient, IQR=28), with 15 suspected ADRs. No correlation was observed between patient severity and ADRs at admission (p=0.26), during hospitalization (p=0.91), or at follow-up (p=0.77). There was also no association between death and ADRs (p=0.28) or worse prognosis and ADRs (p>0.05). Higher BADRI scores correlated with more ADRs (p=0.001). CONCLUSIONS: These data suggest that employing the severity and clinical prognosis scores used in ICUs is not sufficient to direct active pharmacovigilance efforts, which are therefore indicated for critically ill patients.

Effect of pH on the solubility and volumetric change of ready-to-use Bio-C Repair bioceramic material.

Acidic pH can modify the properties of repair cements. In this study, volumetric change and solubility of the ready-to-use bioceramic repair cement Bio-C Repair (BCR, Angelus, Londrina, PR, Brazil) were evaluated after immersion in phosphate-buffered saline (PBS) (pH 7.0) or butyric acid (pH 4.5). Solubility was determined by the difference in initial and final mass using polyethylene tubes measuring 4 mm high and 6.70 mm in internal diameter that were filled with BCR and immersed in 7.5 mL of PBS or butyric acid for 7 days. The volumetric change was established by using bovine dentin tubes measuring 4 mm long with an internal diameter of 1.5 mm. The dentin tubes were filled with BCR at 37°C for 24 hours. Scanning was performed with micro-computed tomography (micro-CT; SkyScan 1176, Bruker, Kontich, Belgium) with a voxel size of 8.74 µm. Then, the specimens were immersed in 1.5 mL of PBS or butyric acid at and 37 °C for 7 days. After this period, a new micro-CT scan was performed. Bio-C Repair showed greater mass loss after immersion in butyric acid when compared with immersion in PBS (p<0.05). Bio-C Repair showed volumetric loss after immersion in butyric acid and increase in volume after immersion in PBS (p<0.05). The acidic pH influenced the solubility and dimensional stability of the Bio-C Repair bioceramic cement, promoting a higher percentage of solubility and decrease in volumetric values.

Factors influencing pregnancy per artificial insemination (AI) and embryonic mortality in Nelore females subjected to timed-AI in Brazil.

A greater understanding of factors influencing fertility is essential to improve pregnancy rates and reduce the occurrence of embryonic mortality in beef herds. The objective of the current study was to evaluate retrospective data of pregnancy per artificial insemination (P/AI) and pregnancy loss in Nelore females subjected to timed-AI (TAI) in Brazil. Data from 40,104 TAI collected from six breeding seasons (2016-2022) were analyzed, and the effects of animal category (e.g., classification based on age and parity), farm, month of parturition, sire, sire breed (Nelore vs Angus), estrus expression at TAI, animal temperament, and body condition scores (BCS) were evaluated. P/AI and pregnancy loss were affected (P < 0.001) by animal category. There was also an effect of farm (P = 0.0013) on P/AI and pregnancy loss (P = 0.001), as P/AI ranged from 49.28% and 55.58% and pregnancy loss from 3.37% to 6.89% across the herds evaluated. Month of parturition also affected (P < 0.001) P/AI and was higher for cows that became pregnant at the beginning of the previous breeding season. Calmer animals, presenting lower velocity scores while exiting the chute following TAI, achieved higher P/AI (P < 0.001). Lower BCS at TAI was associated (P < 0.001) with increased pregnancy loss, and BCS gain following AI was associated (P < 0.001) with reduced rates of embryonic mortality. There was a major effect (P < 0.001) of sire on P/AI and pregnancy loss, as P/AI ranged from 11% to 79%, and embryonic mortality from 0% to 40% for the bulls used in the study, highlighting the importance of the sire fertility on overall pregnancy success. Results from the current study reinforce the idea that animal age and parity at the beginning of the breeding season, BCS at the onset of estrous synchronization, BCS gain following AI, estrus expression at TAI, sire, and month of parturition are important factors influencing P/AI and rates of embryonic mortality in beef herds.

Trabalho interdisciplinar e condições de trabalho de enfermeiros que atuam na atenção primária à saúde / Interdisciplinary work and working conditions of nurses working in primary health care / Trabajo interdisciplinario y condiciones de trabajo de los enfermeros que actúan en la atención primaria de salud

Objetivo: Discutir acerca do trabalho interdisciplinar na prática do enfermeiro na Atenção Primária à Saúde e as suas condições de trabalho. Métodos: Pesquisa descritiva, qualitativa, realizada com 45 profissionais Enfermeiros, que atuam na Atenção Primária à Saúde do estado da Paraíba, de dezembro de 2020 a abril de 2021. A coleta de dados foi guiada por um roteiro semiestruturado. Os dados foram interpretados com base na Análise de Conteúdo e discutidos a partir da literatura existente. Resultados: Evidenciou-se que, apesar de os enfermeiros conseguirem firmar parceria com os outros trabalhadores da unidade, usuários e gestão, e sentirem autonomia para realizar procedimentos, existem condições que dificultam suas práticas, como o pouco reconhecimento profissional, os problemas com a gestão, a falta de insumos e a sobrecarga de função. Conclusão: Os enfermeiros na Atenção Primária vêm buscando seu espaço e melhores condições de trabalho que facilitem suas práticas e favoreçam o cuidado ao usuário. Dessa forma, tais profissionais necessitam de visibilidade e reconhecimento social de suas competências técnicas. (AU)

Objective: Discuss about the interdisciplinary work in the practice of nurses in Primary Health Care and their working conditions. Methods: Descriptive, qualitative research, carried out with 45 professional nurses, who work in Primary Health Care in the state of Paraíba, from December 2020 to April 2021. Data collection was guided by a semi-structured script. Data were interpreted based on Content Analysis and discussed from the existing literature. Results: It was evidenced that, although the nurses manage to establish a partnership with the other workers of the unit, users and management, and feel autonomy to carry out procedures, there are conditions that hinder their practices, such as little professional recognition, problems with management, the lack of inputs and the function overload. Conclusion: Nurses in Primary Care have been seeking their space and better working conditions that facilitate their practices and favor user care. Thus, such professionals need visibility and social recognition of their technical skills. (AU)

Objetivo: Discutir sobre el trabajo interdisciplinario en la práctica de los enfermeros en la Atención Primaria de Salud y sus condiciones de trabajo. Métodos: Investigación descriptiva, cualitativa, realizada con 45 enfermeros profesionales, que actúan en la Atención Primaria de Salud en el estado de Paraíba, de diciembre de 2020 a abril de 2021. La recolección de datos fue guiada por un guión semiestructurado. Los datos fueron interpretados con base en el Análisis de Contenido y discutidos a partir de la literatura existente. Resultados: Se evidenció que, aunque los enfermeros logran establecer una sociedad con los demás trabajadores de la unidad, usuarios y gerencia, y sienten autonomía para realizar los procedimientos, existen condiciones que dificultan sus prácticas, como poco reconocimiento profesional, problemas con la gestión, la falta de insumos y la sobrecarga de funciones. Conclusión: Los enfermeros en Atención Básica vienen buscando su espacio y mejores condiciones de trabajo que faciliten sus prácticas y favorezcan la atención al usuario. Por lo tanto, estos profesionales necesitan visibilidad y reconocimiento social de sus habilidades técnicas. (AU)

Dioclea violacea lectin inhibits tumorigenesis and tumor angiogenesis in vivo.

Dioclea violacea seed mannose-binding lectin (DvL) has attracted considerable attention because of its interesting biological activities, including antitumor, antioxidant, and anti-inflammatory activities. This study evaluated the cytotoxic effect of DvL on tumor and normal cells using the mitochondrial activity reduction (MTT) assay, the carcinogenic and anti-carcinogenic activity by the epithelial tumor test (ETT) in Drosophila melanogaster, and the anti-angiogenic effect by the chick embryo chorioallantoic membrane (CAM) assay. Data demonstrated that DvL promoted strong selective cytotoxicity against tumor cell lines, especially A549 and S180 cells, whereas normal cell lines were weakly affected. Furthermore, DvL did not promote carcinogenesis in D. melanogaster at any concentration tested, but modulated DXR-induced carcinogenesis at the highest concentrations tested. In the CAM and immunohistochemical assays, DvL inhibited sarcoma 180-induced angiogenesis and promoted the reduction of VEGF and TGF-ß levels at all concentrations tested. Therefore, our results demonstrated that DvL is a potent anticancer, anti-angiogenic, and selective cytotoxic agent for tumor cells, suggesting its potential application as a prototype molecule for the development of new drugs with chemoprotective and/or antitumor effects.

Research methods assessing sodium hypochlorite cytotoxicity: A scoping review.

Sodium hypochlorite (NaOCl) cytotoxicity has been assessed using different methodologies, which has led to arbitrary interpretations. This scoping review aimed to discuss the different methodological protocols for assessing NaOCl cytotoxicity. A literature review performed in the PubMed and Embase databases up to July 2023 identified manuscripts reporting NaOCl cytotoxicity. The dataset identified 546 publications, further screened by 2 reviewers. Ninety studies were identified and mined methodologically to collect information on cell type, cytotoxicity assay, NaOCl dilution solutions, presence of fetal bovine serum (FBS), and NaOCl exposure time. The culture medium used in cytotoxicity assays contains buffering substances that neutralize the pH of NaOCl, thus reducing its cytotoxicity, an approach that may lead to bias when solutions with different pH are compared. For short exposure periods, as in simulations to evaluate the contact between irrigant and periapical tissue cells during chemo-mechanical preparation, NaOCl dilution should be performed with saline, which does not buffer the irrigant. For long exposure periods, as in simulations of irrigant extrusions, NaOCl should be diluted in the culture medium, to reproduce the expected buffering effect occurring in extrusions. The presence of FBS in culture medium can decrease NaOCl toxicity. There is no standardization of NaOCl cytotoxicity methodologies. This poses the risk of arriving at incorrect results and, therefore, pertinent tests must be refined.

Cycling using functional electrical stimulation therapy to improve motor function and activity in post-stroke individuals in early subacute phase: a systematic review with meta-analysis.

BACKGROUND: Stroke necessitates interventions to rehabilitate individuals with disabilities, and the application of functional electrical stimulation therapy (FEST) has demonstrated potential in this regard. This study aimed to analyze the efficacy and effectiveness of cycling using FEST to improve motor function and lower limb activity in post-stroke individuals. METHODS: We performed a systematic review according to the recommendations of the PRISMA checklist, searching MEDLINE, Cochrane, EMBASE, LILACS, and PEDro databases by July 2022, without any date or language limitations. Studies were selected using the following terms: stroke, electrical stimulation therapy, cycling, and clinical trials. Randomized or quasi-randomized clinical trials that investigated the effectiveness of cycling using FEST combined with exercise programs and cycling using FEST alone for motor function and activity in subacute post-stroke individuals were included. The quality of included trials was assessed using the PEDro scores. Outcome data were extracted from eligible studies and combined in random-effects meta-analyses. The quality of evidence was determined according to the Grading of Recommendations Assessment, Development, and Evaluation system. RESULTS: Five randomized clinical trials involving 187 participants were included. Moderate-quality evidence indicates that cycling using FEST combined with exercise programs promotes relevant benefits in trunk control (MD 9 points, 95% CI 0.36-17.64) and walking distance (MD 94.84 m, 95% CI 39.63-150.05, I = 0%), the other outcomes had similar benefits. Cycling using FEST alone compared to exercise programs promotes similar benefits in strength, balance, walking speed, walking distance, and activities of daily living. CONCLUSION: This systematic review provides low- to moderate-quality evidence that cycling using FEST may be an effective strategy to consider in improving motor function and activity outcomes for post-stroke individuals in the early subacute phase. REVIEW REGISTRATION: PROSPERO (CRD42022345282).

Density-based lipoprotein depletion improves extracellular vesicle isolation and functional analysis.

BACKGROUND: Blood plasma is the main source of extracellular vesicles (EVs) in clinical studies aiming to identify biomarkers and to investigate pathophysiological processes, especially regarding EV roles in inflammation and thrombosis. However, EV isolation from plasma has faced the fundamental issue of lipoprotein contamination, representing an important bias since lipoproteins are highly abundant and modulate cell signaling, metabolism, and thromboinflammation. OBJECTIVES: Here, we aimed to isolate plasma EVs after depleting lipoproteins, thereby improving sample purity and EV thromboinflammatory analysis. METHODS: Density-based gradient ultracentrifugation (G-UC) was used for lipoprotein depletion before EV isolation from plasma through size-exclusion chromatography (SEC) or serial centrifugation (SC). Recovered EVs were analyzed by size, concentration, cellular source, ultrastructure, and bottom-up proteomics. RESULTS: G-UC efficiently separated lipoproteins from the plasma, allowing subsequent EV isolation through SEC or SC. Combined analysis from EV proteomics, cholesterol quantification, and apoB-100 detection confirmed the significant reduction in lipoproteins from isolated EVs. Proteomic analysis identified similar gene ontology and cellular components in EVs, regardless of lipoprotein depletion, which was consistent with similar EV cellular sources, size, and ultrastructure by flow cytometry and transmission electron microscopy. Importantly, lipoprotein depletion increased the detection of less abundant proteins in EV proteome and enhanced thromboinflammatory responses of platelets and monocytes stimulated in vitro with EV isolates. CONCLUSION: Combination of G-UC+SEC significantly reduced EV lipoprotein contamination without interfering in EV cellular source, gene ontology, and ultrastructure, allowing the recovery of highly pure EVs with potential implications for functional assays and proteomic and lipidomic analyses.

Inhibition of Mitochondrial Translation Ameliorates Imiquimod-Induced Psoriasis-Like Skin Inflammation by Targeting Vγ4+ γδ T Cells.

Psoriasis is an inflammatory skin disorder that is characterized by keratinocyte hyperproliferation in response to immune cell infiltration and cytokine secretion in the dermis. γδ T cells expressing the Vγ4 TCR chain are among the highest contributors of IL-17A, which is a major cytokine that drives a psoriasis flare, making Vγ4+ γδ T cells a suitable target to restrict psoriasis progression. In this study, we demonstrate that mitochondrial translation inhibition within Vγ4+ γδ T cells effectively reduced erythema, scaling, and skin thickening in a murine model of psoriatic disease. The antibiotic linezolid, which blocks mitochondrial translation, inhibited the production of mitochondrial-encoded protein cytochrome c oxidase in Vγ4+ γδ T cells and systemically reduced the frequencies of IL-17A+ Vγ4+ γδ T cells, effectively resolving IL-17A-dependent inflammation. Inhibiting mitochondrial translation could be a novel metabolic approach to interrupt IL-17A signaling in Vγ4+ T cells and reduce psoriasis-like skin pathophysiology.