EQA total haemoglobine quantification: long-term analytical performance for thirty Portuguese Laboratories

Total haemoglobin is one of the most frequently ordered laboratory tests, both in the hospital and ambulatory. Haemoglobin concentration provides information about the status of anaemia in the population and guides many clinical diagnoses and therapeutic interventions. Portuguese External Quality Evaluation Program (PNAEQ) provides 4 survey per year with 2 samples each since it is important for medical laboratories to know about the stability and performance of their methods over the time. The main objective of this study was to apply a linear regression model for long-term evaluation of the performance of laboratories in haemoglobin quantification by comparing the laboratory individual results with the consensus mean of each round, after outliers exclusion. To evaluate the performance of the laboratory, the long-term analytical CV (LCVa) and the total analytical bias were established.N/

International Normalized Ratio (INR): Performance of External Quality Assessment 2016 results - PNAEQ and ECAT Foundation

The International Normalized Ration (INR) is derived from the measurement of the Prothrombin Time (PT) and the International Sensitivity Index (ISI), and is a quantitative measure of the responsiveness of individual thromboplastin reagents to the different clotting factors involved in the PT measurement. Under the INR system, all results are standardized. For example, a person taking an oral anticoagulant (ex. Coumadin) would need regularly to have a blood test to measure the INR. The INR permits patients on anticoagulants to travel and obtain comparable test results wherever they are. So it is very important to verify the laboratories performance over this test. In 2014 the Portuguese National External Quality Assessment Program (PNAEQ) has established a consortium with ECAT Foundation for INR measurement where Portuguese laboratories are included in the statistical analysis. The main objective of this study was to evaluate the performance for INR of PNAEQ participants that used two different thromboplastin reagents during 2016 and compared those with the results of ECAT participants.N/

DECISÕES DO GOVERNO BRASILEIRO NO ENFRENTAMENTO DA COVID-19: REPERCUSSÕES PARA O CUIDADO CLÍNICO DE ENFERMAGEM

Objective: to analyze the repercussions of the decisions of the Brazilian federal government proposed to face COVID-19 in clinical nursing care. Method: documentary study based on publications from January 2020 to June 2021 published on the official website of the federal government. A total of 144 documents were obtained, which were analyzed in full, presented in a flowchart and Descending Hierarchical Classification (CHD), the latter generated by the IRaMuTeQ Software. Results: CHD generated six classes: social scope, public management, economics in the pandemic and health care targeting. Decisions focused on recognizing the existence of the pandemic scenario and establishing the creation of committees for management and confrontation, management of hospital supplies, redefinition of goals related to health indicators and combating violence. Final considerations: the determinations to face the pandemic presented actions limited, primarily, to the curative aspects that impacted the offer of clinical nursing care by reinforcing the biomedical model of health care.Objetivo: analizar las repercusiones de las decisiones del gobierno federal brasileño propuestas para enfrentar la COVID-19 en la atención clínica de enfermería. Método: estudio documental basado en publicaciones de enero de 2020 a junio de 2021 publicadas en el sitio web oficial del gobierno federal. Se obtuvo un total de 144 documentos, los cuales fueron analizados en su totalidad, presentados en un diagrama de flujo y Clasificación Jerárquica Descendente (CHD), esta última generada por el Software IRaMuTeQ. Resultados: la CHD generó seis clases: alcance social, gestión pública, economía en la pandemia y focalización de la atención en salud. Las decisiones se centraron en reconocer la existencia del escenario de pandemia y establecer la creación de comités de gestión y enfrentamiento, gestión de insumos hospitalarios, redefinición de metas relacionadas con indicadores de salud y combate a la violencia. Consideraciones finales: las determinaciones para enfrentar la pandemia presentaron acciones limitadas, principalmente, a los aspectos curativos que impactaron la oferta de cuidados clínicos de enfermería reforzando el modelo biomédico de atención a la salud.Objetivo: analisar as repercussões das decisões do Governo Federal brasileiro no enfrentamento da COVID-19 no cuidado clínico de enfermagem. Método: estudo documental baseado nas publicações de janeiro de 2020 a junho de 2021 divulgadas no site oficial do governo federal. Obtiveram-se 144 documentos que foram analisados na íntegra, apresentados em fluxograma e em Classificação Hierárquica Descendente, esta gerada pelo software IRaMuTeQ. Resultados: a Classificação Hierárquica Descendente gerou seis classes condensadas em: COVID-19 no Brasil, âmbito social, gestão pública, economia na pandemia e direcionamento da assistência em saúde. As decisões centraram-se em reconhecer a existência do cenário pandêmico e estabelecer criação de comitês para gestão e enfrentamento, gestão de insumos hospitalares, redefinição de metas relacionadas aos indicadores de saúde e combate à violência. Considerações finais: as determinações para enfrentamento da pandemia apresentaram ações circunscritas, prioritariamente aos aspectos curativistas, que impactaram na oferta dos cuidados clínicos de enfermagem ao reforçarem o modelo biomédico de atenção à saúde. Descritores: Governo. Pandemias. COVID-19. Políticas. Cuidado de Enfermagem

Perfis de DPOC e características tratáveis utilizando recursos mínimos: Identificação, árvore de decisão e estabilidade longitudinal

Background: Chronic obstructive pulmonary disease (COPD) is highly heterogeneous and complex. Hence, personalising assessments and treatments to this population across different settings and available resources imposes challenges and debate. Research efforts have been made to identify clinical phenotypes or profiles for prognostic and therapeutic purposes. Nevertheless, such profiles often do not describe treatable traits, focus on complex physiological/pulmonary measures which are frequently not available across settings, lack validation and/or their stability over time is unknown. Objective: To identify profiles and their treatable traits based on simple and meaningful measures; to develop and validate a profile decision tree; and to explore profiles’ stability over time in people with COPD. Methods: An observational, prospective study was conducted with people with COPD. Clinical characteristics, lung function, symptoms, impact of the disease (COPD assessment test–CAT), health-related quality of life, physical activity, lower-limb muscle strength and functional status were collected cross-sectionally and a subsample was followed-up monthly over six months. A principal component analysis and a clustering procedure with k-medoids were applied to identify profiles. Pulmonary and extrapulmonary (i.e., physical, symptoms and health status, and behavioural/life-style risk factors) treatable traits were identified in each profile based on the established cut-offs for each measure available in the literature. The decision tree was developed with 70% and validated with 30% of the sample, cross-sectionally. Agreement between the profile predicted by the decision tree and the profile defined by the clustering procedure was determined using Cohen’s Kappa. Stability was explored over time with a stability score defined as the percentage ratio between the number of timepoints that a participant was classified in the same profile (most frequent profile for that participant) and the total number of timepoints (i.e., 6). Results: 352 people with COPD (67.4±9.9 years; 78.1% male; FEV1=56.2±20.6% predicted) participated and 90 (67.6±8.9 years; 85.6% male; FEV1=52.1±19.9% predicted) were followed-up. Four profiles were identified with distinct treatable traits. The decision tree was composed by the CAT, age and FEV1% predicted and had an agreement of 71.7% (Cohen’s Kappa=0.62, p<0.001) with the actual profiles. 48.9% of participants remained in the same profile whilst 51.1% moved between two (47.8%) and three (3.3%) profiles over time. The overall stability of profiles was 86.8±15%. Conclusion: Profiles and treatable traits can be identified in people with COPD with simple and meaningful measures possibly available even in minimal-resource settings. Regular assessments are recommended as people with COPD may change profile over time and hence their needs of personalised treatment.N/

Clinical characteristics of COPD Phenotypes

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COPD profiles and treatable traits using minimal resources: identification, decision tree and stability over time

Background and objective: Profiles of people with chronic obstructive pulmonary disease (COPD) often do not describe treatable traits, lack validation and/or their stability over time is unknown. We aimed to identify COPD profiles and their treatable traits based on simple and meaningful measures; to develop and validate a decision tree and to explore profile stability over time. Methods: An observational, prospective study was conducted. Clinical characteristics, lung function, symptoms, impact of the disease (COPD Assessment Test—CAT), health-related quality of life, physical activity, lower-limb muscle strength and functional status were collected cross-sectionally and a subsample was followed-up monthly over six months. A principal component analysis and a clustering procedure with k-medoids were applied to identify profiles. A decision tree was developed and validated cross-sectionally. Stability was explored over time with the ratio between the number of timepoints that a participant was classified in the same profile and the total number of timepoints (i.e., 6). Results: 352 people with COPD (67.4±9.9 years; 78.1% male; FEV1=56.2±20.6% predicted) participated and 90 (67.6±8.9 years; 85.6% male; FEV1=52.1±19.9% predicted) were followed-up. Four profiles were identified with distinct treatable traits. The decision tree included CAT (<18 or≥18 points); age (<65 or≥65 years) and FEV1 (<48 or≥48% predicted) and had an agreement of 71.7% (Cohen’s Kappa=0.62, p<0.001) with the actual profiles. 48.9% of participants remained in the same profile whilst 51.1% moved between two (47.8%) or three (3.3%) profiles over time. Overall stability was 86.8±15%. Conclusion: Four profiles and treatable traits were identified with simple and meaningful measures possibly available in low-resource settings. A decision tree with three commonly used variables in the routine assessment of people with COPD is now available for quick allocation to the identified profiles in clinical practice. Profiles and treatable traits may change over time in people with COPD hence, regular assessments to deliver goal-targeted personalised treatments are needed.publishe

Volatilomics reveals potential biomarkers for identification of renal cell carcinoma: an in vitro approach

The identification of noninvasive biomarkers able to detect renal cell carcinoma (RCC) at an early stage remains an unmet clinical need. The recognition that altered metabolism is a core hallmark of cancer boosted metabolomic studies focused in the search for cancer biomarkers. The present work aims to evaluate the performance of the volatile metabolites present in the extracellular medium to discriminate RCC cell lines with distinct histological subtypes (clear cell and papillary) and metastatic potential from non-tumorigenic renal cells. Hence, volatile organic compounds (VOCs) and volatile carbonyl compounds (VCCs) were extracted by headspace solid-phase microextraction (HS-SPME) and analyzed by gas chromatography-mass spectrometry (GC-MS). Multivariate and univariate analysis unveiled a panel of metabolites responsible for the separation between groups, mostly belonging to ketones, alcohols, alkanes and aldehydes classes. Some metabolites were found similarly altered for all RCC cell lines compared to non-tumorigenic cells, namely 2-ethylhexanol, tetradecane, formaldehyde, acetone (increased) and cyclohexanone and acetaldehyde (decreased). Furthermore, significantly altered levels of cyclohexanol, decanal, decane, dodecane and 4-methylbenzaldehyde were observed in all metastatic RCC cell lines when compared with the non-metastatic ones. Moreover, some alterations in the volatile composition were also observed between RCC histological subtypes. Overall, our results demonstrate the potential of volatile profiling for identification of noninvasive candidate biomarkers for early RCC diagnosis.info:eu-repo/semantics/publishedVersio

Sirtuins’ deregulation in bladder cancer: SIRT7 is implicated in tumor progression through epithelial to mesenchymal transition promotion

Sirtuins are emerging players in cancer biology and other age-related disorders, and their putative role in bladder cancer (BlCa) remains elusive. Further understanding of disease biology may allow for generation of more effective pathway-based biomarkers and targeted therapies. Herein, we aimed to illuminate the role of sirtuins’ family in BlCa and evaluate their potential as disease biomarkers and therapeutic targets. SIRT1-7 transcripts and protein levels were evaluated in a series of primary BlCa and normal bladder mucosa tissues. SIRT7 knockdown was performed through lentiviral transduction in MGHU3, 5637 and J82 cells and its functional role was assessed. SIRT1, 2, 4 and 5 expression levels were significantly lower in BlCa, whereas SIRT6 and 7 were overexpressed, and these results were corroborated by TCGA cohort analysis. SIRT7 transcript levels were significantly decreased in muscle-invasive vs. papillary BlCa. In vitro studies showed that SIRT7 downregulation promoted cells migration and invasion. Accordingly, increased EMT markers expression and decreased E-Cadherin (CDH1) was observed in those BlCa cells. Moreover, increased EZH2 expression and H3K27me3 deposition in E-Cadherin promoter was found in sh-SIRT7 cells. We demonstrated that sirtuins are globally deregulated in BlCa, and specifically SIRT7 downregulation is implicated in EMT, fostering BlCa invasiveness through EZH2-CDH1 axis.This research was supported by the Research Center of the Portuguese Oncology Institute of Porto (CI-IPOP–FBGEBC-27 and PI 74-CI-IPOP-19-2016), by Fundação para a Ciência e Tecnologia (FCT) (PhD fellowships SFRH/BD/112673/2015 to S.M.-R and SFRH/BD/92786/2013 to C.S.G.; IF/00601/2012 to B.M.C.), and by Fundo Europeu de Desenvolvimento Regional (FEDER) (post-doctoral fellowships IPO/ESTIMANORTE01-0145-FEDER-000027 to V.M.-G. and COMPETE/FEDER/FCT_CI-IPOP-BPD/UID/DTP/00776/2013 to I.G.)

Identification of Marker Compounds and In Vitro Toxicity Evaluation of Two Portuguese Asphodelus Leaf Extracts

This article belongs to the Special Issue Discovery of Bioactive Ingredients from Natural Products III.The leaves of Asphodelus bento-rainhae subsp. bento-rainhae, an endemic Portuguese species, and Asphodelus macrocarpus subsp. macrocarpus have been used as food, and traditionally as medicine, for treating ulcers, urinary tract, and inflammatory disorders. The present study aims to establish the phytochemical profile of the main secondary metabolites, together with the antimicrobial, antioxidant and toxicity assessments of both Asphodelus leaf 70% ethanol extracts. Phytochemical screenings were conducted by the TLC and LC-UV/DAD-ESI/MS chromatographic technique, and quantification of the leading chemical classes was performed by spectrophotometric methods. Liquid-liquid partitions of crude extracts were obtained using ethyl ether, ethyl acetate, and water. For in vitro evaluations of antimicrobial activity, the broth microdilution method, and for the antioxidant activity, the FRAP and DPPH methods were used. Genotoxicity and cytotoxicity were assessed by Ames and MTT tests, respectively. Twelve known compounds including neochlorogenic acid, chlorogenic acid, caffeic acid, isoorientin, p-coumaric acid, isovitexin, ferulic acid, luteolin, aloe-emodin, diosmetin, chrysophanol, and β-sitosterol were identified as the main marker compounds, and terpenoids and condensed tannins were found to be the major class of secondary metabolites of both medicinal plants. The ethyl ether fractions demonstrated the highest antibacterial activity against all the Gram-positive microorganisms, (MIC value of 62 to 1000 µg/mL), with aloe-emodin as one of the main marker compounds highly active against Staphylococcus epidermidis (MIC value of 0.8 to 1.6 µg/mL). Ethyl acetate fractions exhibited the highest antioxidant activity (IC50 of 800 to 1200 µg/mL, respectively). No cytotoxicity (up to 1000 µg/mL) or genotoxicity/mutagenicity (up to 5 mg/plate, with/without metabolic activation) were detected. The obtained results contribute to the knowledge of the value and safety of the studied species as herbal medicines.This research was funded by the Foundation for Science and Technology (FCT, Portugal) through national funds FCT/MCTES to iMed.ULisboa (UIDP/04138/2020, UIDB/04138/2020) and MEtRICs (UIDP/04077/2020, UIDB/04077/2020) research projects, as well as doctoral scholarship (SFRH/BD/125310/2016) granted to the first author.info:eu-repo/semantics/publishedVersio