O Essencial da Transferência: Uma nova (re)visão dos preditores da Transferência da Formação

Tese de Doutoramento em Políticas de Desenvolvimento de Recursos HumanosAtualmente, a transferência da formação encontra-se numa encruzilhada. As reconhecidas vantagens da formação surgem a par da dificuldade em quantificar o grau de transferência para o contexto de trabalho. A literatura tem convidado a um novo debate sobre o que já se sabe sobre o sistema de transferência da formação e os fatores que a influenciam. Pretende revisitar-se um conjunto de antecedentes da transferência, considerando alguns cuja relação já está validada na literatura, como os fatores do “Learning Transfer System Inventory” (LTSI) ou as estratégias de transferência, outros cuja investigação em torno desta relação é ainda incipiente, como os comprometimentos profissional e organizacional, e outros ainda que não foram estudados em associação com a transferência, como os estilos de tomada de decisão. A presente investigação propõe considerar estes constructos como antecedentes da transferência, perspetivando a hipótese dos comprometimentos, os estilos de tomada de decisão e as estratégias de transferência surgirem como mediadores na relação entre os preditores do LTSI e a transferência da formação. Esta investigação é composta por quatro estudos assentes numa abordagem hipotético-dedutiva e construtivista, de natureza quantitativa, com recurso à metodologia de inquérito por questionário. Os modelos concetuais foram testados através da modelação por equações estruturais. Consoante os estudos, foram também analisados os efeitos de moderação e mediação presentes, bem como a identificação de perfis associados à transferência da formação. Os resultados finais evidenciaram a emergência da motivação para transferir, coaching de desempenho e estratégias de transferência cognitivo-afetivas como antecedentes, estas últimas identificadas como mediador da relação entre aqueles e a transferência da formação. A Gestão e a teoria beneficiam da identificação dos antecedentes mais impactantes no âmbito do sistema de transferência, essencial para políticas formativas mais eficazes e para um adequado redirecionamento do investimento em formação. Além desta conclusão geral, são apontadas outras conclusões e sugestões para futuras investigações.Nowadays, training transfer is at a conceptual and practical crossroad. The recognized advantages of training appear alongside the difficulty in quantifying the degree of training transfer that happens in the workplace. Literature has opened a new debate about what is already known about this transfer system and the factors that influence it. We intend to revisit a set of training transfer antecedents, considering some whose relationship is already widely validated in the literature, such as the factors in the “Learning Transfer System Inventory” (LTSI) or the transfer strategies. Also, we intend to include others whose relationship with transfer is still incipient, as the professional and organizational commitments, and others that have not been studied in association with transfer, as decisionmaking styles. This investigation presents these constructs as antecedents of training transfer, suggesting that both commitments, decision-making styles, and transfer strategies act as mediating variables in the relationship between the predictors of LTSI and training transfer. This investigation consists of four studies based on a hypothetical-deductive and constructivist approach, of a quantitative nature, using the questionnaire methodology. Conceptual models are tested through Structural Equation Modeling (SEM). Depending on the studies, the moderation and mediation effects are also analyzed, as well as the identification of profiles associated with training transfer. The results highlighted the emergence of motivation to transfer, performance coaching and cognitive-affective transfer strategies as the sole antecedents of training transfer, with these strategies showing a mediating role between those constructs and training transfer. Human Resources Management and literature benefit from the identification of the most impactful antecedents within the transfer system, essential for the design of more effective policies and for an adequate redirection of investment in training. In addition to this general conclusion, several other conclusions and suggestions for future investigations are pointed out in each study.N/

New Insights on Innate Immune Response by Blood Macrophages and Liver Kupffer Cells to Leishmania infantum Parasites

Funding Information: Funding: This study was supported by FCT-Foundation for Science and Technology, I.P., through research grant PTDC/CVT-CVT/28908/2017 and PTDC/CVT-CVT/0228/2020, and by national funds within the scope of Centro de Investigação Interdisciplinar em Sanidade Animal (CIISA, UIDB/00276/2020) and Global Health and Tropical Medicine (GHTM, UID/04413/2020). Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.L. infantum is the aetiological agent of zoonotic visceral leishmaniasis (ZVL), a disease that affects humans and dogs. Leishmania parasites are well adapted to aggressive conditions inside the phagolysosome and can control the immune activation of macrophages (MØs). Although MØs are highly active phagocytic cells with the capacity to destroy pathogens, they additionally comprise the host cells for Leishmania infection, replication, and stable establishment in the mammal host. The present study compares, for the first time, the innate immune response to L. infantum infection of two different macrophage lineages: the blood macrophages and the liver macrophages (Kupffer cells, KC). Our findings showed that L. infantum takes advantage of the natural predisposition of blood-MØs to phagocyte pathogens. However, parasites rapidly subvert the mechanisms of MØs immune activation. On the other hand, KCs, which are primed for immune tolerance, are not extensively activated and can overcome the dormancy induced by the parasite, exhibiting a selection of immune mechanisms, such as extracellular trap formation. Altogether, KCs reveal a different pattern of response in contrast with blood-MØs when confronting L. infantum parasites. In addition, KCs response appears to be more efficient in managing parasite infection, thus contributing to the ability of the liver to naturally restrain Leishmania dissemination.publishersversionpublishe

Immunophenotyping of peripheral blood, lymph node, and bone marrow T lymphocytes during canine leishmaniosis and the impact of antileishmanial chemotherapy

Dogs are a major reservoir of Leishmania infantum, etiological agent of canine leishmaniosis (CanL) a zoonotic visceral disease of worldwide concern. Therapeutic protocols based on antileishmanial drugs are commonly used to treat sick dogs and improve their clinical condition. To better understand the impact of Leishmania infection and antileishmanial drugs on the dog's immune response, this study investigates the profile of CD4+ and CD8+ T cell subsets in peripheral blood, lymph node, and bone marrow of sick dogs and after two different CanL treatments. Two CanL groups of six dogs each were treated with either miltefosine or meglumine antimoniate combined with allopurinol. Another group of 10 clinically healthy dogs was used as control. Upon diagnosis and during the following 3 months of treatment, peripheral blood, popliteal lymph node, and bone marrow mononuclear cells were collected, labeled for surface markers CD45, CD3, CD4, CD8, CD25, and intracellular nuclear factor FoxP3, and T lymphocyte subpopulations were immunophenotyped by flow cytometry. CanL dogs presented an overall increased frequency of CD8+ and CD4+CD8+ double-positive T cells in all tissues and a decreased frequency of CD4+ T cells in the blood. Furthermore, there was a higher frequency of CD8+ T cells expressing CD25+FoxP3+ in the blood and bone marrow. During treatment, these subsets recovered to levels similar to those of healthy dogs. Nevertheless, antileishmanial therapy caused an increase of CD4+CD25+FoxP3+ T cells in all tissues, associated with the decrease of CD8+CD25−FoxP3− T cell percentages. These findings may support previous studies that indicate that L. infantum manipulates the dog's immune system to avoid the development of a protective response, ensuring the parasite's survival and the conditions that allow the completion of Leishmania life cycle. Both treatments used appear to have an effect on the dog's immune response, proving to be effective in promoting the normalization of T cell subsets.publishersversionpublishe

immune activation of canine hepatic spheroids exposed to leishmania infantum

The application of innovative three-dimensional (3D) spheroids cell culture strategy to Parasitology offers the opportunity to closely explore host–parasite interactions. Here we present a first report on the application of 3D hepatic spheroids to unravel the immune response of canine hepatocytes exposed to Leishmania infantum. The liver, usually considered a major metabolic organ, also performs several important immunological functions and constitutes a target organ for L. infantum infection, the etiological agent of canine leishmaniasis (CanL), and a parasitic disease of major veterinary and public health concern. 3D hepatic spheroids were able to sense and immunologically react to L. infantum parasites, generating an innate immune response by increasing nitric oxide (NO) production and enhancing toll-like receptor (TLR) 2 and interleukin-10 gene expression. The immune response orchestrated by canine hepatocytes also lead to the impairment of several cytochrome P450 (CYP450) with possible implications for liver natural xenobiotic metabolization capacity. The application of meglumine antimoniate (MgA) increased the inflammatory response of 3D hepatic spheroids by inducing the expression of Nucleotide oligomerization domain (NOD)-like receptors 1 and NOD2 and TLR2, TLR4, and TLR9 and enhancing gene expression of tumour necrosis factor α. It is therefore suggested that hepatocytes are key effector cells and can activate and orchestrate the immune response to L. infantum parasites.publishersversionpublishe

Phenotypic Features of Circulating Leukocytes from Non-human Primates Naturally Infected with Trypanosoma cruzi Resemble the Major Immunological Findings Observed in Human Chagas Disease

Background: Cynomolgus macaques (Macaca fascicularis) represent a feasible model for research on Chagas disease since natural T. cruzi infection in these primates leads to clinical outcomes similar to those observed in humans. However, it is still unknown whether these clinical similarities are accompanied by equivalent immunological characteristics in the two species. We have performed a detailed immunophenotypic analysis of circulating leukocytes together with systems biology approaches from 15 cynomolgus macaques naturally infected with T. cruzi (CH) presenting the chronic phase of Chagas disease to identify biomarkers that might be useful for clinical investigations. Methods and findings: Our data established that CH displayed increased expression of CD32+ and CD56+ in monocytes and enhanced frequency of NK Granzyme A+ cells as compared to non-infected controls (NI). Moreover, higher expression of CD54 and HLA-DR by T-cells, especially within the CD8+ subset, was the hallmark of CH. A high level of expression of Granzyme A and Perforin underscored the enhanced cytotoxicity-linked pattern of CD8+ T-lymphocytes from CH. Increased frequency of B-cells with up-regulated expression of Fc-γRII was also observed in CH. Complex and imbricate biomarker networks demonstrated that CH showed a shift towards cross-talk among cells of the adaptive immune system. Systems biology analysis further established monocytes and NK-cell phenotypes and the T-cell activation status, along with the Granzyme A expression by CD8+ T-cells, as the most reliable biomarkers of potential use for clinical applications. Conclusions: Altogether, these findings demonstrated that the similarities in phenotypic features of circulating leukocytes observed in cynomolgus macaques and humans infected with T. cruzi further supports the use of these monkeys in preclinical toxicology and pharmacology studies applied to development and testing of new drugs for Chagas disease

Meglumine antimoniate and miltefosine combined with allopurinol sustain pro-inflammatory immune environments during canine leishmaniosis treatment

Canine leishmaniosis (CanL) caused by Leishmania infantum is a zoonotic disease of global concern. Antileishmanial drug therapies commonly used to treat sick dogs improve their clinical condition, although when discontinued relapses can occur. Thus, the current study aims to evaluate the effect of CanL treatments in peripheral blood, lymph node, and bone marrow cytokine profile associated with clinical recovery. Two groups of six dogs diagnosed with CanL were treated with miltefosine combined with allopurinol and meglumine antimoniate combined with allopurinol (MT+A and MG+A), respectively. At diagnosis and after treatment, during a 3-month follow-up, clinical signs, hematological and biochemical parameters, urinalysis results and antileishmanial antibody titers were registered. Furthermore, peripheral blood, popliteal lymph node, and bone marrow samples were collected to assess the gene expression of IL-2, IL-4, IL-5, IL-10, IL-12, TNF-α, TGF-β, and IFN-γ by qPCR. In parallel, were also evaluated samples obtained from five healthy dogs. Both treatment protocols promoted the remission of clinical signs as well as normalization of hematological and biochemical parameters and urinalysis values. Antileishmanial antibodies returned to non-significant titers in all dogs. Sick dogs showed a generalized upregulation of IFN-γ and downregulation of IL-2, IL-4, and TGF-β, while gene expression of IL-12, TNF-α, IL-5, and IL-10 varied between groups and according to evaluated tissue. A trend to the normalization of cytokine gene expression was induced by both miltefosine and meglumine antimoniate combined therapies. However, IFN-γ gene expression was still up-regulated in the three evaluated tissues. Furthermore, the effect of treatment in the gene expression of cytokines that were not significantly changed by infection, indicates that miltefosine and meglumine antimoniate combined therapy directly affects cytokine generation. Both combined therapies are effective in CanL treatment, leading to sustained pro-inflammatory immune environments that can compromise parasite survival and favor dogs' clinical cure. In the current study, anti-inflammatory and regulatory cytokines do not seem to play a prominent role in CanL or during clinical recovery.publishersversionpublishe

Meglumine antimoniate and miltefosine combined with allopurinol sustain pro-inflammatory immune environments during canine leishmaniosis treatment

Research Areas: Veterinary SciencesCanine leishmaniosis (CanL) caused by Leishmania infantum is a zoonotic disease of global concern. Antileishmanial drug therapies commonly used to treat sick dogs improve their clinical condition, although when discontinued relapses can occur. Thus, the current study aims to evaluate the effect of CanL treatments in peripheral blood, lymph node, and bone marrow cytokine profile associated with clinical recovery. Two groups of six dogs diagnosed with CanL were treated with miltefosine combined with allopurinol and meglumine antimoniate combined with allopurinol (MT+A and MG+A), respectively. At diagnosis and after treatment, during a 3-month follow-up, clinical signs, hematological and biochemical parameters, urinalysis results and antileishmanial antibody titers were registered. Furthermore, peripheral blood, popliteal lymph node, and bone marrow samples were collected to assess the gene expression of IL-2, IL-4, IL-5, IL-10, IL-12, TNF-alpha, TGF-beta, and IFN-gamma by qPCR. In parallel, were also evaluated samples obtained from five healthy dogs. Both treatment protocols promoted the remission of clinical signs as well as normalization of hematological and biochemical parameters and urinalysis values. Antileishmanial antibodies returned to non-significant titers in all dogs. Sick dogs showed a generalized upregulation of IFN-gamma and downregulation of IL-2, IL-4, and TGF-beta, while gene expression of IL-12, TNF-alpha, IL-5, and IL-10 varied between groups and according to evaluated tissue. A trend to the normalization of cytokine gene expression was induced by both miltefosine and meglumine antimoniate combined therapies. However, IFN-gamma gene expression was still up-regulated in the three evaluated tissues. Furthermore, the effect of treatment in the gene expression of cytokines that were not significantly changed by infection, indicates that miltefosine and meglumine antimoniate combined therapy directly affects cytokine generation. Both combined therapies are effective in CanL treatment, leading to sustained pro-inflammatory immune environments that can compromise parasite survival and favor dogs' clinical cure. In the current study, anti-inflammatory and regulatory cytokines do not seem to play a prominent role in CanL or during clinical recovery.info:eu-repo/semantics/publishedVersio

Programa educativo em medidas de precaução universais: uma metodologia de abordagem

An updating program on measures of universal precautions (M.U.P.) was developed at the Center of Whole Care of Woman's Health (Centro de Atenção Integral à Saúde da Mulher - CAISM). These measures and the procedures in the case of work accident were published in a booklet. First, servants should be aware of the matter of stress and its influence on the quality of life. Then, updating was carried through encouraging the reflection on the consequences of the non-adoption of M.U.P. The answers to 286 pre-tests and 242 post-tests were analyzed and the results showed a significantly higher index of correct answers (p< 0,01), mainly regarding the appropriate use of glove.El Centro de Atención Integral a la Salud de la Mujer (CAISM) desarrolló un programa de reciclaje sobre Medidas de Precaución Universales (MPU). Se divulgó por medio de un material educativo (cartilla) las MPU y procedimientos en caso de accidentes de trabajo. En primer lugar se cuestionó al funcionário sobre el stress y su influencia en la calidad de vida, llevando se a cabo el reciclaje en el cual se promovió la reflexión sobre consecuencias cuando las MPU no son adoptadas. Fueron respondidas y analizadas 286 pre-test y 242 pos-test con un índice de acierto significativamente mayor en este último (p < 0,01), principalmente con relación al uso adecuado de los guantes.No Centro de Atenção Integral à Saúde da Mulher (CAISM) desenvolveu-se um programa de reciclagem sobre as Medidas de Precaução Universais (MPU) e divulgou-se, através de uma cartilha, essas medidas e os procedimentos em caso de acidente de trabalho. Primeiramente, o funcionário era sensibilizado para a questão do estresse e sua influência sobre a qualidade de vida e, após, era realizada a reciclagem promovendo-se a reflexão sobre as conseqüências da não adoção das MPU. Foram respondidos e analisados 286 pré-testes e 242 pós-testes com um índice de acertos significativamente maior nestes últimos (

Construção de um plano de formação numa instituição particular de solidariedade social

Este estudo decorre do Plano Estratégico da Instituição na qual foi reconhecida a necessidade de conceber um Plano de Formação para os colaboradores, estando este trabalho direcionado ao grupo de educadoras das respostas sociais da Creche e Jardim de Infância. A formação é um contributo fundamental para a qualidade da Instituição. As mudanças na sociedade são constantes e os profissionais devem estar preparados e motivados para dar resposta às suas exigências. Os recursos humanos são a essência da Instituição e a sua implicação e vontade são fundamentais para a sua qualidade. Neste projeto, recorremos a uma metodologia de natureza qualitativa, elegendo como técnicas de recolha de dados: a entrevista, a análise documental e o focus group. Com base nos resultados obtidos, e considerando a especificidade da Instituição, concebemos um Plano de Formação para o triénio 2018-2020: procedimentos, intervenientes, instrumentos de avaliação e plano de melhoria. Palavras-chave: Formação profissional; desenvolvimento profissional; capital humano; qualidade da organização; desempenho organizacional; formação continua; necessidades de formação.This study stems from the Institution’s Strategic Plan in which it was recognized the need to generate an Instruction Plan for its collaborators. This project is specifically aimed at the group of educators from both Day Care and Kindergarten’s social responses. Instruction plays a major role as far as the Institution’s quality is concerned. Society is constantly changing and every professional must be prepared and motivated to meet its demands and requirements. Human resources are the Institution’s essence and with that being said, its implication and will are fundamental to its quality. In this project, we decided to use a qualitative methodology, choosing interview, documental analisis and focus group as our data collection technique. Based on our results, and taking into account the specificity of the institution, we have conceived an Instruction Plan aiming the 2018-2020 triennium: procedures, intervenients, evaluation tools and improvement plan. Key Words: Professional Instruction, Professional Development, Human capital, Organization’s quality, Organizational performance; continuous instruction; training needs

Canine leishmaniasis: the role of hepatocytes and Kupffer cells during Leishmania infantum infection

A leishmaniose canina (LCan) é uma doença zoonótica causada por Leishmania infantum, um parasita protozoário transmitido por flébotomos. O fígado constitui um órgão vital e de grande importância metabólica, sendo também um alvo preferencial para L. infantum. Apesar do complexo metabolismo, o fígado apresenta mecanismos efetores que podem contribuir para a direta eliminação do parasita de forma ainda não completamente compreendida. Assim, este estudo tem por objetivo analisar o papel dos hepatócitos e das células de Kupffer (CK) na infeção por L. infantum. A resposta imune orquestrada pelo cão contra L. infantum é constituída por um largo espetro de respostas de imunidade inata e adquirida. Estudos em cães infetados demonstraram que a imunidade ao parasita é órgão-específica. Para analisar a interação entre L. infantum e os hepatócitos, no presente estudo, hepatócitos e CK foram isolados de cães saudáveis e cultivadas em dois sistemas de cultura diferentes. O sistema 2D consiste na clássica cultura em placa e o sistema 3D tira partido da tendência dos hepatócitos para agregar e permite às células recriar a organização natural existente no órgão. L. infantum apresentou um claro tropismo para os hepatócitos, aderindo fortemente à membrana celular. No entanto, não é claro se o parasita entrou nos hepatócitos. Estas interações levaram a uma ativação da imunidade inata, aumentando a expressão génica dos receptores NOD1 e NOD2, bem como TLR2, em sinergia com a diferenciação de um perfil misto de citocinas anti- e pro-inflamatórias, que conduziu à diminuição de atividade das CYPs450. Três novos compostos com reconhecida atividade anti-leishmanial (ácido ursólico, chalcona-8 e quercetina) foram testados e comparados com o fármaco classicamente utilizado no tratamento da leishmaniose. Os compostos experimentais apresentaram boa atividade leishmanicida e não afetaram a viabilidade dos hepatócitos, diminuíram a inflamação celular e restauraram a atividade das CTPs450, havendo ainda a hipótese de serem metabolizados em parte pelas referidas enzimas. As CK mostraram-se permissivas a amastigotas e promastigotas e elevaram a expressão de NOD1 e TLR2 na presença do parasita. No entanto, L. infantum é capaz de regular a resposta imunitária nas CK, evitando a ação de citocinas anti e pró-inflamatórias e do stress oxidativo, aumentando, por outro, lado a síntese de ureia. A adição de antimoniato de meglumina permitiu às CK recuperar a ativação e expressão de recetores de imunidade inata e produção de citocinas. A co-cultura de CK infetadas com hepatócitos revelou que as células em conjunto foram capazes de ativar a imunidade inata. Os hepatócitos evidenciaram, assim, um papel interessante na orquestração de uma resposta imune sinergética com as CK contra L. infantum. Foram ainda observadas diferenças entre a resposta imune exibida por macrófagos do sangue (MØ) e CK infetados por L. infantum. A análise da memória linfocitária hepática, realizada em modelo murino de leishmaniose visceral, demonstrou que o tratamento com antimoniato de meglumina elevou os níveis de memória e o número de células efetoras, melhorando a resposta imunitária do fígado, salientando o seu papel como órgão imunitário. As proteínas recombinantes de L. infantum, LirCyp1 e LirSOD foram reconhecidas pelas células de memória de animais infetados e tratados, indiciando que estas proteínas podem vir a ser incorporadas numa vacina terapêutica ou profilática.Canine leishmaniasis (CanL) is a zoonotic disease caused by Leishmania infantum, a protozoan parasite transmitted by sand flies. The liver is a vital and major metabolic organ which also constitutes a L. infantum preferential target. Despite the complex metabolism, liver possesses effector mechanisms that can contribute to direct elimination of Leishmania in a not yet defined way. Thus, this study aimed to assess the role of hepatocytes and Kupffer cells (KC) in the canine liver infected by L. infantum. A wide spectrum of innate and acquired immune responses is orchestrated by the canine host when facing L. infantum infection. CanL infection studies have indicated that parasite immunity is organ-specific. In the present study, canine hepatocytes and KCs were isolated from healthy stray dogs and two different culture systems were used to analyze the interaction of L. infantum with hepatocytes. The 2D-culture system consists in the classical plate culture and the 3D-culture system takes advantage of the natural aggregation properties of hepatocytes and allows cells to recreate some of the natural tissue architecture. L. infantum exhibited evident tropism to hepatocytes and strongly attached to the cells in both cultures system. However, it was not clear if parasites entered the hepatocytes. These interactions lead to activation of innate immunity by increasing the gene expression of NOD1 and NOD2 receptors in synergy with the increase in TLR2 expression. TNF-α cytokine expression was increased creating a proinflammatory cellular microenvironment and, as consequence, decreasing the CYP450 activity. This downregulation of CYPs may increase toxicity of clinical drugs in some cases. Three new potential leishmanicidal drugs were tested, ursolic acid, chalcone-8 and quercetin in comparison to the standard meglumine antimoniate. Experimental drugs revealed good leishmanicidal activity and did not reduce hepatocyte viability. The addition of leishmanicidal drugs decreased the inflammation and restored the CYP450 activity, which were also probable involved in the metabolization of the tested compounds. Regarding KCs, these cells were permissive to L. infantum amastigotes and promastigotes. KCs increased expression of NOD1 and TLR2 innate immune receptors. Even so, the parasite regulates KC immune response, avoiding the generation of proand anti-inflammatory cytokines and the oxidative burst, increasing the synthesis of urea. The addition of meglumine antimoniate allowed KC to activate, increasing the expression of innate immune receptors and of pro-inflammatory cytokines, breaking the silencing imposed by the parasite. The addition of L. infantum infected KCs to canine hepatocytes in a co-culture, was able to activate cells innate immunity. Hepatocyte seemed to have an interesting role in orchestrating a synergistic immune response against L. infantum parasites. Major differences were also found between the immune response exhibited by blood derivate macrophages (MØ) and liver KC, when facing infection by L. infantum. The assessment of the liver lymphocyte memory performed in the murine model of visceral leishmaniasis demonstrated that the treatment with antimoniate meglumine increases the levels of memory and effector T cells, enhancing liver immune response. The liver has higher levels of immune memory T cells, evidencing its role as an immunological organ. The recombinant proteins LirCyp1 and LirSOD, are strongly recognized by memory cells from infected and treated mice, indicating these proteins might be used in a prophylactic or therapeutic vaccine formulation