Development of a new approach based on NADH dehydrogenase subunit 1(ND1) marker and high resolution melting (HRM) analysis towards the authentication of the geographical origin of honey

In the present study, an approach for verifying the geographical origin of honey based on its entomological origin is proposed. The method was developed based on the polymerase chain reaction (PCR) amplification of a new markercontaining different single nucleotide polymorphisms characteristic of honeybees of different mitochondrial (mtDNA) lineages, therefore generating different fluorescent curves on HRM analysis. The method was successfully applied to honeys from Portugal and Italy, expected to be from lineages A and C, respectively, demonstrating their origin compliance.The authors are grateful to Programa Nacional Apícola 2020-2022 for funding the project “AUTENT+ Desenvolvimento de abordagens inovadoras com vista à valorização e exploração do potencial de mercado do mel Português”, to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO UIDB/00690/2020, to Fenapícola and Capemel for supplying the Portuguese honeys and to Dr. Antonio Nanetti (CREA-AA) for the Italian honeys. D. Henriques is supported by the project Bee Happy (POCI-01-0145-FEDER-029871) funded by FEDER, COMPETE 2020-POCIand FCT and A.R. Lopes by the PhD scholarship funded by the FCT SFRH/BD/143627/2019.info:eu-repo/semantics/publishedVersio

Suppression by allogeneic-specific regulatory T cells is dependent on the degree of HLA compatibility

Copyright © 2021 The Authors. This article is distributed under the terms of the CC BY 4.0 Unported license.Regulatory T cell (Treg) infusion for graft-versus-host disease treatment has been increasingly investigated. However, polyclonal Treg may suppress the desired graft-versus-leukemia effect. Although allogeneic-specific (allo-specific) Treg may provide a more-targeted graft-versus-host disease treatment, there is the need to develop easily translatable expansion protocols and to better characterize their specificity and mechanisms of suppression. In this article, we provide a robust protocol for human allo-specific Treg expansion and characterize their phenotype, potency, and specificity of suppression by testing different expansion conditions and suppression assay milieus. We found that higher concentrations of IL-2 during expansion with allogeneic APC yielded allo-specific Treg that were more-potent suppressors and displayed a more activated phenotype. Although responses to the same APC present during expansion were the most suppressed, responses to third-party APC partially matched to the expansion APC were still significantly more suppressed than responses to fully mismatched APC. Furthermore, suppression of responses to the expansion APC was strictly contact dependent, whereas suppression of responses to mismatched APC was partially independent of contact. Finally, distinct subsets in fresh and expanded Treg could be described using multidimensional visualization techniques. We propose that allo-specific Treg are HLA specific and that the mechanisms of suppression elicited depend on their compatibility with the stimulators.This work was supported by Fundação para a Ciência e Tecnologia, Portugal, under the Harvard Medical School–Portugal Program Project Induction of Immune Tolerance in Human Allogeneic Hematopoietic Stem Cell Transplantation (HMSP-ICT/0001/2011) and Gilead Sciences, under Programa Gilead GÉNESE (FPJ001262). J.B. received a Ph.D. grant from Fundação para a Ciência e Tecnologia (PD/BD/105769/2014).info:eu-repo/semantics/publishedVersio

Patient-centeredness: contribution to the adaptation of the Patient-Practitioner Orientation Scale (PPOS)

Objetivo: Este estudo teve como objetivo traduzir e contribuir para a adaptação para a população portuguesa (Português Europeu) da Patient-Practitioner Orientation Scale (PPOS). Método: Após o processo de tradução e de pré-teste, a escala foi aplicada a 593 estudantes do 1º ao 6º ano do curso de Medicina em várias Universidades de Portugal Continental. A validade do construto e a fiabilidade do instrumento foram aferidas através da análise fatorial exploratória (ACP) e confirmatória (AFC), e do cálculo do coeficiente alpha de Cronbach. Resultados: A versão final explica 31.54% da variância total e confirma a estrutura em dois fatores: Caring, (19.56% da variância) e Sharing (11.98% da variância). Os itens 2 e 4 apresentaram inconsistências com os fatores definidos à priori (versão original do instrumento), os itens 9 e 17 obtiveram cargas fatoriais inferiores a .3, e o item 3 registou uma diferença inferior a .1 entre as cargas fatoriais para os dois domínios. Os coeficientes de alpha de Cronbach foram .65, .50 e .56 para a escala total, e subscalas Caring e Sharing, respetivamente. A AFC revelou um bom ajustamento global do modelo de medida (χ2(132, N = 593) = 344.28, p < .001; χ2/gl = 2.61; GFI = .93; AGFI = .92; CFI = .87; NNFI = .81; SRMR = .084; RMSEA = .05, 95% CI [0.045, 0.059], p = .293). As análises exploratórias posteriores sugerem a possibilidade de melhoria dos índices de validade e de fiabilidade da escala total e da sub-escala Caring, com a retirada de itens específicos. Conclusão: Não obstante as fragilidades encontradas no que concerne à fiabilidade e validade da PPOS-P para uma amostra de estudantes portugueses de Medicina, este estudo representa um contributo científico para a adaptação da escala, que pode ser considerada para efeitos de avaliação de atitudes de centração no paciente nos contextos da educação médica e da investigação.ABSTRACT - Objective: The purpose of this study was to contribute to the European Portuguese adaptation of the Patient-Practitioner Orientation Scale (PPOS). Method: A sample of 593 medical students participated in the study. After permission from the original author, the translation procedures required to ensure translation of the PPOS to European Portuguese were performed. Construct validity (exploratory and confirmatory factor analysis) and reliability (internal consistency) were assessed. Results: The final version confirmed the original structures of two factors, explaining 31.54% of total variance; Caring (19.56%) and Sharing (11.98%). Items 2 and 4 showed inconsistencies with the factors defined earlier in the original version of the instrument, items 9 and 17 obtained a factorial load less than .3, and the item 3 achieved a difference of less than .1 between factorial charges for the two domains. The internal consistency of PPOS-P scales was adequate (Cronbach’s alpha of .65, .50 and .56 for total scale, and subscales Caring and Sharing, respectively). Confirmatory factor analysis provided an acceptable adjustment for the observed variables (χ2(132, N = 593) = 344.28, p < .001; χ2/gl = 2.61; GFI = .93; AGFI = .92; CFI = .87; NNFI = .81; SRMR = .084; RMSEA = .05, 95% CI [0.045, 0.059], p = .293). Subsequent exploratory analyzes suggest the potential for improving the levels of validity and reliability of the total scale and Caring subscale, with the removal of specific items. Conclusion: Although the fragilities identified in the validity and reliability of the PPOS-P in a sample of Portuguese medical students, this work can represent an important and useful contribution to further investigations that might consider this instrument as a measure of student’s changes of patient-centeredness attitudes.info:eu-repo/semantics/publishedVersio

A mixed-methods approach

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Organizational and job resources on employees' job insecurity during the first wave of COVID-19: the mediating effect of work engagement

The world of work has been severely affected by the COVID-19 pandemic due to the high instability observed in the labor market, bringing several new challenges for leaders and employees. The present study aims to analyze the role of organizational and job resources in predicting employees’ job insecurity during the first wave of the COVID-19 outbreak, through the mediating role of work engagement. A sample of 207 Portuguese employees participated (Mean age = 45 years old, SD = 9.92), of which 64.7% were women. Data was collected using an online survey, including self-report measures of organizational resources (perceived organizational support), job resources (performance feedback and job autonomy), job insecurity, and work engagement. Data showed that job and organizational resources negatively influenced job insecurity. Moreover, work engagement was a significant mediator of the relation between performance feedback (facet of job resources) and job insecurity. Findings suggest that investing in job and organizational resources can act as protective factors to minimize feelings of job insecurity. Likewise, leaders should foster work engagement among employees to help them balance the relation between these resources and job insecurity, especially in crisis situations. Overall, this study takes a new, underexplored perspective, theoretically bridging organizational and job resources with job insecurity and work engagement during a time of great uncertainty, such as the COVID-19 pandemic

Clarifying changes in student empathy throughout medical school: a scoping review

Despite the increasing awareness of the relevance of empathy in patient care, some findings suggest that medical schools may be contributing to the deterioration of students' empathy. Therefore, it is important to clarify the magnitude and direction of changes in empathy during medical school. We employed a scoping review to elucidate trends in students' empathy changes/differences throughout medical school and examine potential bias associated with research design. The literature published in English, Spanish, Portuguese and French from 2009 to 2016 was searched. Two-hundred and nine potentially relevant citations were identified. Twenty articles met the inclusion criteria. Effect sizes of empathy scores variations were calculated to assess the practical significance of results. Our results demonstrate that scoped studies differed considerably in their design, measures used, sample sizes and results. Most studies (12 out of 20 studies) reported either positive or non-statistically significant changes/differences in empathy regardless of the measure used. The predominant trend in cross-sectional studies (ten out of 13 studies) was of significantly higher empathy scores in later years or of similar empathy scores across years, while most longitudinal studies presented either mixed-results or empathy declines. There was not a generalized international trend in changes in students' empathy throughout medical school. Although statistically significant changes/differences were detected in 13 out of 20 studies, the calculated effect sizes were small in all but two studies, suggesting little practical significance. At the present moment, the literature does not offer clear conclusions relative to changes in student empathy throughout medical school.info:eu-repo/semantics/publishedVersio

Against all odds: a tale of marine range expansion with maintenance of extremely high genetic diversity

The displacement of species from equatorial latitudes to temperate locations following the increase in sea surface temperatures is among the significant reported consequences of climate change. Shifts in the distributional ranges of species result in fish communities tropicalisation, i.e., high latitude colonisations by typically low latitude distribution species. These movements create new interactions between species and new trophic assemblages. The Senegal seabream, Diplodus bellottii, may be used as a model to understand the population genetics of these invasions. In the last decades, this species has undergone an outstanding range expansion from its African area of origin to the Atlantic coast of the Iberian Peninsula, where now occurs abundantly. Mitochondrial and nuclear markers revealed a striking high haplotypic nucleotide and genetic diversity values, along with significant population differentiation throughout the present-day geographical range of the Senegal seabream. These results are not consistent with the central-marginal hypothesis, nor with the expectations of a leptokurtic distribution of individuals, as D. bellottii seems to be able to retain exceptional levels of diversity in marginal and recently colonised areas. We discuss possible causes for hyperdiversity and lack of geographical structure and subsequent implications for fisheries.Portuguese Foundation for Science and Technology: UID/MAR/04292/2019/ UID/Multi/04326/2019info:eu-repo/semantics/publishedVersio

Pleistocene expansion, anthropogenic pressure and ocean currents: Disentangling the past and ongoing evolutionary history of Patella aspera Röding, 1798 in the archipelago of Madeira

Aims: Rising sea-level following the Last Glacial Maximum lead to fragmentation of coastal limpet populations between islands of the Archipelago of Madeira. This fragmentation is reinforced by recent heavy exploitation reducing effective population size on Madeira Island. We use the limpet P. aspera to understand how the role of processes at different time scales (i.e. changes in the sea level and overexploitation) can influence the genetic composition of an extant species, relating these processes to reproductive phenology and seasonal shifts in ocean currents. Location: Madeira Island, Porto Santo and Desertas (Archipelago of Madeira, NE Atlantic Ocean). Taxon: The limpet Patella aspera. Methods: Twelve microsatellite genetic markers were used. A power analysis was used to evaluate the power of the microsatellite markers to detect a signal of population differentiation. Long-term past migrations were assessed using a Bayesian Markov Montecarlo approach in the software MIGRATE-n to estimate mutation-scaled migration rates (M = m/μ; m, probability of a lineage immigrating per generation; μ, mutation rate). Two sce narios were evaluated using an Approximate Bayesian Computation (ABC) in the software DIYABC 2.1 (i) Sce nario 1: considered a population scenario from a reduced Ne at time t3 to a higher Ne at time t2; and (ii) Scenario 2 considering a reduction of Ne from a time t3 to a time t2. Results: Colonization of the archipelago by Portuguese settlers six centuries ago probably led to an important decrease in the genetic diversity of the species (Ne). Contemporary gene flow strongly support a pattern of high asymmetric connectivity explained by the reproductive phenology of the species and spatio-temporal seasonal changes in the ocean currents. Spatio-temporal reconstructions using Bayesian methods, including coalescent and Approximate Bayesian Computation (ABC) approaches, suggest changes in the migration patterns from highly symmetric to highly asymmetric connectivity with subtle population differentiation as consequence of post-glacial maximum sea level rise during the Holocene.info:eu-repo/semantics/publishedVersio

Aplasia Cutis Congenita: An Often Isolated Clinical Finding

Aplasia cutis congenita (ACC) is a rare congenital condition characterised by the absence of skin, with or without the absence of underlying structures. Can appear as an isolated lesion or associated with other congenital malformations. The approach can be conservative or surgical. We report a case of a full-term male newborn with two adjacent ulcerated lesions with well-demarcated limits in the upper occipital region (20 and 4-5 mm diameter). Other alterations that could be associated with aplasia cutis were identified at examination, so the patient was referred to several consultations, and other system involvement was excluded. Re-evaluation at the first month of life revealed complete healing and conservative treatment was kept. After identification of ACC should be excluded other associated congenital malformations. Particularly if large skin defects or bone involvement is detected (higher risk of complications and mortality)