Otimização da caracterização de fibras musculares estriadas em cão (Canis familiaris) por histoquímica e imunohistoquímica com potencial aplicação ao diagnóstico

Dissertação de Mestrado Integrado em Medicina VeterináriaAs técnicas histoquímicas permitem melhorar o conhecimento sobre o tecido muscular e suas alterações patológicas e, juntamente com as técnicas imunohistoquímicas, têm papel importante no diagnóstico de doenças musculares. Assim, o presente trabalho teve como objetivos, em primeiro lugar, caracterizar o tecido muscular esquelético da espécie canina, tanto a nível histoquímico como imunohistoquímico, incidindo, sobretudo, na tipificação e subtipificação das fibras musculares e avaliação das suas percentagens relativas. Em segundo lugar, pretendeu-se verificar a potencial aplicação de técnicas imunohistoquímicas no diagnóstico, aplicando-as em músculos alterados. As técnicas histoquímicas realizadas foram: adenosina trifosfatase ácido resistente (ATPase 4,6), adenosina trifosfatase alcalina (ATPase 9,4) e desidrogenase tetrazolium redutase do ácido nicotínico (NADH diaforase). Estas foram efetuadas em cortes de congelação dos músculos temporal, masséter, diafragma e bicípede femoral de dois canídeos eutanasiados sem história de doença muscular. Os mesmos músculos destes canídeos foram também submetidos a imunohistoquímica, recorrendo aos anticorpos MHC-slow e MHC-fast. Posteriormente, estes anticorpos foram aplicados em três biópsias musculares de canídeos previamente diagnosticados com doença muscular. Dos resultados obtidos pela histoquímica foi possível diferenciar fibras de tipo I e II, subtipificar as fibras tipo II e obter a percentagem de fibras tipo I e II nos diferentes músculos. Com a imunohistoquímica, registou-se a percentagem de células marcadas pelos anticorpos atrás referidos e a intensidade de cada marcação, consoante o músculo ou o caso clínico em questão. Não foi possível observar a existência de correlação entre as fibras de tipo I e II marcadas pela histoquímica e a marcação obtida por imunohistoquímica, não existindo igualmente correlação entre as percentagens de fibras marcadas pelos dois tipos de técnicas. Finalmente, foram também identificadas as condições necessárias e suficientes para a realização das técnicas de diagnóstico de patologia muscular no Laboratório de Anatomia Patológica da Faculdade de Medicina Veterinária da Universidade de Lisboa (FMV-ULisboa), reconhecendo-se as limitações e a forma de as ultrapassar.ABSTRACT - Optimization of the Characterization of Striated Muscle Fibers in Dog (Canis familiaris) by Histochemistry and Immunohistochemistry with Possible Application in the Diagnosis. - Histochemical techniques are able to increase the comprehension regarding normal and diseased muscular tissue and, together with immunohistochemical techniques, they have an important role in the diagnosis of muscular diseases. Therefore, the present work aimed, primarily, to characterize dog’s musculoskeletal tissue, at the histochemical and immunohistochemical level, focusing, essentially, on the typification and subtypification of muscle fibers. Secondly, it was intended to apply immunohistochemical techniques to altered muscle tissue, in order to determine if these techniques can be used in the diagnosis of dog muscular diseases. The following histochemical techniques were performed: adenosine triphosphatase at pH 4.6 (ATPase 4.6), adenosine triphosphatase at pH 9.4 (ATPase 9.4) and reduced nicotinamide adenine dinucleotide-tetrazolium redutase (NADH diaphorase). These techniques were performed in frozen sections of the temporalis, masseter, diaphragm and biceps femoris muscles that belonged to two dogs euthanized with no history of muscular disease. The same muscles were also submitted to immunohistochemical techniques, using the antibodies MHC-slow and MHC-fast. These antibodies were also applied to muscle biopsies of three dogs diagnosed with muscular disease. Histochemistry made possible to differentiate type I and II fibers, to identify the subtypes of type II fibers and to determine the percentage of type I and II fibers in the different muscles. Immunohistochemistry, made possible to determine the percentage of marked cells by both antibodies. The intensity of stain was registered according to the muscle or the clinical case analyzed. However, it was not possible to obtain any correlation between the percentage of type I and II fibers marked by histochemistry and by immunohistochemistry or no correlation was found between the percentages of muscle fibers marked by both techniques. Finally, the present study also allowed the identification of the conditions necessary and sufficient to perform these techniques in the diagnosis of muscle pathology to be put up in the Laboratory of Anatomical Pathology of the Faculty of Veterinary Medicine (University of Lisbon), recognizing its limitations and how to overcome them

The mediating role of self-criticism, experiential avoidance and negative urgency on the relationship between ED-related symptoms and difficulties in emotion regulation

Objective: Difficulties in emotion regulation are thought to play a transdiagnostic role across eating disorders (ED). In the current study, we explored with a path analysis the mediating role of self-criticism, experiential avoidance and negative urgency on the relationship between ED-related symptoms and dimensions of difficulties in emotion regulation. Method: Participants were 103 female outpatients recruited at a Portuguese ED hospital unit, diagnosed with an ED, aged 14–60 years old (M = 28.0, SD = 10.5), body mass index (BMI) ranging from 11.72 to 39.44 (M = 20.1, SD = 5.4). Results: The path analysis resulted in a model with an adequate fit to the data (SRMR = 0.05; RMSEA = 0.07 [0.00, 0.12], PCLOSE = 0.269; TLI = 0.97; IFI = 0.99; GFI = 0.95). A final model in which the relationship between ED-related symptoms and dimensions of difficulties in emotion regulation was mediated by self-criticism, experiential avoidance and negative urgency, accounted for a variance of 71% for strategies, 57% for non-acceptance, 62% for impulses, 56% for goals and 20% for clarity. Conclusion: Results suggest that self-criticism, experiential avoidance and negative urgency, combined, are relevant in the relationship between ED-related symptoms and difficulties in emotion regulation. ED treatment and emotion regulation skills may be enhanced through the inclusion of specific components that target self-criticism, experiential avoidance and negative urgency, as they become prominent during the therapeutic process.FCT - Fundação para a Ciência e a Tecnologia(POCI‐01‐0145‐FEDER‐028145

The Lockdown Impact on the Relations between Portuguese Parents and Their 1- to 3-Year-Old Children during the COVID-19 Pandemic

Funding Information: In Portugal, during the first lockdown, with few exceptions, families with children under 12 years old were given a choice. One parent could suspend his/her professional activity temporarily and receive financial support from the National Social Security System []. Most families, especially those with little children, chose this option. Publisher Copyright: © 2022 by the authors.Many countries have applied mandatory confinement measures in response to the COVID-19 pandemic, such as school and kindergarten closures, which confined families to their homes. The study concerns the impacts of the first COVID-19 lockdown on the relationships between Portuguese parents and their children, in a non-clinical population composed of fathers and mothers of children between the ages of 12 months and 3 years and 364 days. An online questionnaire (set by the research team) and the Parenting Daily Hassles Scale (PDHS) concerning the confinement period were applied between 17 June and 29 July 2020. To assess the impacts of the lockdown, outcomes regarding the impacts perceived by the parents, the potential regression in the development of children, and the willingness to promote changes in family routines in the future, were considered. Of the total sample (n = 1885), 95.4% of the parents (n = 1798) said that, after confinement, the relationship with their children had improved or remained similar to the pre-confinement period; 97.3% (n = 1835) noticed positive changes in the development of their children, and 63.7% (n = 1200) noted that the relationships with their children during the confinement period would lead to some changes in family routines in the future. Multivariate regression analyses showed that most of the sociodemographic variables chosen were not associated with the outcomes. However, significant levels of pressure over parenting and parental overload (reported by high scores in the PDHS intensity and frequency scales), challenging behaviors of the children, and the impacts they had on parental tasks had negative influences on the studied outcomes. On the contrary, the number of adults living with their children, the perceptions regarding the development of their children, and sharing new experiences with them were significant factors for positively-perceived impacts on the relationships between them or in the desire to bring about changes in family routines in the future. The impacts of the lockdown on the relationships between parents and children aged between 1 and 3 years old were more dependent on relational aspects and on the parents’ sense of competence in exercising parental functions. We conclude that, despite the increased demands imposed by the lockdown, nearly all of the parents evaluated the quality of the relationship with their children as positive after this period.publishersversionpublishe

Impact of COVID-19 lockdown measures on a cohort of eating disorders patients

BackgroundLockdown implemented to prevent the COVID-19 spread resulted in marked changes in the lifestyle. The objective of the current study was to assess the impact of lockdown measures on a cohort of eating disorder (ED) patients being followed as part of an ongoing naturalistic treatment study.MethodsNinety-nine patients aged 18 or older, currently or previously, in treatment at a Portuguese specialized hospital unit were contacted by phone and invited to participate in the current survey. Fifty-nine agreed to be interviewed by phone, and 43 agreed to respond to a set of self-report measures of ED symptoms, emotion regulation difficulties, clinical impairment, negative urgency, and COVID-19 impact, during the week after the end of the lockdown period.ResultsData showed that of the 26 patients currently in treatment: 8 remained unchanged (31%), 7 deteriorated (27%), and 11 reliably improved (42%). Of the 17 participants not currently in treatment: 3 deteriorated (18%), 9 remained unchanged (53%), and 5 (29%) improved after the lockdown period. The Coronavirus Impact Scale showed that most patients considered their routines moderately or extremely impacted, experienced stress related to coronavirus, and showed difficulty in maintaining physical exercise and feeding routines. Results suggest that higher impact of COVID-19 lockdown was significantly correlated with eating disorder symptoms and associated psychopathology, impulsivity, difficulties in emotion regulation and clinical impairment measured at post-lockdown. In addition, the impact of COVID-19 and lockdown measures on clinical impairment was mediated by difficulties in emotion regulation.ConclusionsFindings suggest that some ED patients may experience worsening of their condition, especially if associated with difficulties in emotion regulation, and these difficulties might be exacerbated in the context of a stressful crisis and lockdown measures, highlighting the need for intervention strategies to mitigate its negative impact.This work was conducted at the Psychology Research Centre (PSI/01662), School of Psychology, University of Minho, and partially supported by the Portuguese Foundation for Science and Technology and the Portuguese Ministry of Science, Technology and Higher Education (UID/PSI/01662/2019), through the national funds (PIDDAC); and a grant by the Portuguese Foundation for Science and Technology and the Portuguese Ministry of Science, Technology and Higher Education through national funds and cofinanced by FEDER through COMPETE2020 under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-028145) to P. Machado PI)

Antibacterial action mechanisms of honey: physiological effects of avocado, chestnut, and polyfloral honey upon Staphylococcus aureus and Escherichia coli

Numerous studies have explored the antibacterial properties of di erent types of honey from all around the world. However, the data available describing how honey acts against bacteria are few. The aim of this study was to apply a flow cytometry (FC) protocol to examine and characterize the primary e ects of three varieties of honey (avocado, chestnut and polyfloral) upon physiological status of Staphylococcus aureus and Escherichia coli cells to reveal their antibacterial action mechanisms. The e ects of honey samples on membrane potential, membrane integrity, and metabolic activity were assessed using di erent fluorochromes, in a 180 min time course assay. Time-kill experiments were also carried out under similar conditions. Exposure of S. aureus and E. coli to the distinct honey samples resulted in physiological changes related to membrane polarization and membrane integrity. Moreover, honey induced a remarkable metabolic disruption as primary physiological e ect upon S. aureus. The di erent honey samples induced quite similar e ects on both bacteria. However, the depth of bacteria response throughout the treatment varied depending on the concentration tested and among honey varieties, probably due to compositional di erences in the honey.This work was supported by the Consejería de Sanidad of Junta de Castilla y León; under Grant GRS 551/A/10. Patricia Combarros-Fuertes was funded by Consejería de Educación of Junta de Castilla y León and European Social Fund.info:eu-repo/semantics/publishedVersio

Evaluation of physiological effects induced by manuka honey upon staphylococcus aureus and escherichia coli

Several studies have explored the antimicrobial properties of manuka honey (MkH). However, the data available regarding antibacterial action mechanisms are scarcer. The aim of this study was to scrutinize and characterize primary e ects of manuka honey (MkH) upon the physiological status of Staphylococcus aureus and Escherichia coli (as Gram-positive and Gram-negative bacteria models, respectively), using flow cytometry (FC) to reveal its antibacterial action mechanisms. E ects of MkH on membrane potential, membrane integrity and metabolic activity were assessed using di erent fluorochromes in a 180 min time course assay. Time-kill experiments were carried out under the same conditions. Additionally, MkH e ect on e ux pumps was also studied in an E. coli strain with an over-expression of several e ux pumps. Exposure of bacteria to MkH resulted in physiological changes related to membrane potential and membrane integrity; these e ects displayed slight di erences among bacteria. MkH induced a remarkable metabolic disruption as primary physiological effect upon S. aureus and was able to block effux pump activity in a dose-dependent fashion in the E. coli strain.This work was supported by the Consejería de Sanidad of Junta de Castilla y León; under grant GRS 551/A/10. Patricia Combarros-Fuertes was funded by Consejería de Educación of Junta de Castilla y León and European Social Fund.info:eu-repo/semantics/publishedVersio

Educational practices: interactions, challenges and possibilities in the daily life of a Farm Family School

Este artigo apresenta análise das práticas pedagógicas estruturadas pelos Monitores/as que trabalham com o Ensino Médio/Técnico de uma Escola Família Agrícola (EFA), tendo como objetivo evidenciar, através das percepções destes sujeitos sobre suas práticas, desafios e confrontos vivenciados no cotidiano dos espaços de aula, cujo projeto educativo alicerça-se na Formação em Alternância, com princípios pedagógicos/metodológicos diferenciados. De cunho qualitativo, a contribuição desta análise oportunizará aos Monitores/as retomada de reflexão sobre suas práticas, sem pretensão de impor, mas de problematizar, qualificando cada vez mais o processo de elaboração e reelaboração de conhecimentos à luz do projeto educativo da alternância. Sob a orientação de autores atentos ao debate sobre Educação do Campo e Formação em Alternância, percebemos que muitos são os fatores que interferem no fazer pedagógico, onde os Monitores/as reconhecem no contexto educativo a inserção de aspectos políticos, sociais, culturais, religiosos, econômicos e a influência destes na organização das práticas pedagógicas. Provocam desafios e suscitam novas possibilidades a serem trilhadas para a consolidação das práticas, cujas exigências reveladas têm como indicativo estabelecer uma aprendizagem que, de fato, se torne significativa e emancipatória para os sujeitos que dela se apropriam. Palavras-chave: Práticas Pedagógicas, Formação em Alternância, Confrontos e Desafios.   Educational practices: interactions, challenges and possibilities in the daily life of a Farm Family School ABSTRACT. This article presents analysis of pedagogical practices structured by monitors / those working with high school / technical a Family Farm School (EFA), aiming to show, through the perceptions of these subjects about their practices, challenges and experienced clashes in routine of classroom spaces, whose educational project is founded on Training in Alternation with teaching / different methodological principles. Qualitative nature, the contribution of this analysis will provide to the Monitor / the resumption of reflection on their practices, without pretension to impose, but to problematize, describing increasingly the process of elaboration and re-elaboration of knowledge in the light of the educational project of alternation. Under the guidance of authors aware of the debate on Education Course and Training in Alternation, we realize that there are many factors that interfere with pedagogical where monitors/recognize them in an educational context the inclusion of political, social, cultural, religious, economic and their influence in the organization of pedagogical practices. Cause challenges and raise new possibilities to be threshed to the consolidation of practices, whose requirements have revealed as indicative establish a fact that learning becomes meaningful and emancipatory for the subjects that it was appropriated. Keywords: Teaching Practices, Training in Alternation, Clashes and Challenges.   Las prácticas educativas: las interacciones, los retos y posibilidades en la vida cotidiana de una escuela granja de la familia RESUMEN. En este artículo se presenta el análisis de las prácticas pedagógicas estructuradas por monitores / los que trabajan con la escuela secundaria / técnica una Escuela Familiar Agraria (EFA), con el objetivo de mostrar, a través de las percepciones de estos sujetos sobre sus prácticas, desafíos y enfrentamientos con experiencia en rutinario de los espacios del aula, cuyo proyecto educativo se basa en la formación en alternancia con la enseñanza/diferentes principios metodológicos. Naturaleza cualitativa, la contribución de este análisis oportunizará la pantalla / la reanudación de la reflexión sobre sus prácticas, sin pretensión de imponer, sino para problematizar, describiendo cada vez más el proceso de elaboración y reelaboración del conocimiento a la luz del proyecto educativo de la alternancia. Bajo la guía de autores consciente del debate en curso para la formación en alternancia, nos damos cuenta de que hay muchos factores que interfieren con la pedagógica donde los monitores / reconocerlos en un contexto educativo la inclusión de políticas, sociales, culturales, religiosos, económico y su influencia en la organización de las prácticas pedagógicas. Causar problemas y plantear nuevas posibilidades para ser trilladas a la consolidación de prácticas, cuyas necesidades han revelado como indicativo establecer un hecho de que el aprendizaje se vuelve significativa y emancipadora para los sujetos que las había asignado. Palabras-clave: Prácticas de Enseñanza, La Formación en Alternancia; Los Enfrentamientos y Desafíos.  ABSTRACT. This article presents analysis of pedagogical practices structured by monitors / those working with high school / technical a Family Farm School (EFA), aiming to show, through the perceptions of these subjects about their practices, challenges and experienced clashes in routine of classroom spaces, whose educational project is founded on Training in Alternation with teaching / different methodological principles. Qualitative nature, the contribution of this analysis will provide to the Monitor / the resumption of reflection on their practices, without pretension to impose, but to problematize, describing increasingly the process of elaboration and re-elaboration of knowledge in the light of the educational project of alternation. Under the guidance of authors aware of the debate on Education Course and Training in Alternation, we realize that there are many factors that interfere with pedagogical where monitors/recognize them in an educational context the inclusion of political, social, cultural, religious, economic and their influence in the organization of pedagogical practices. Cause challenges and raise new possibilities to be threshed to the consolidation of practices, whose requirements have revealed as indicative establish a fact that learning becomes meaningful and emancipatory for the subjects that it was appropriated.RESUMEN. En este artículo se presenta el análisis de las prácticas pedagógicas estructuradas por monitores / los que trabajan con la escuela secundaria / técnica una Escuela Familiar Agraria (EFA), con el objetivo de mostrar, a través de las percepciones de estos sujetos sobre sus prácticas, desafíos y enfrentamientos con experiencia en rutinario de los espacios del aula, cuyo proyecto educativo se basa en la formación en alternancia con la enseñanza/diferentes principios metodológicos. Naturaleza cualitativa, la contribución de este análisis oportunizará la pantalla / la reanudación de la reflexión sobre sus prácticas, sin pretensión de imponer, sino para problematizar, describiendo cada vez más el proceso de elaboración y reelaboración del conocimiento a la luz del proyecto educativo de la alternancia. Bajo la guía de autores consciente del debate en curso para la formación en alternancia, nos damos cuenta de que hay muchos factores que interfieren con la pedagógica donde los monitores / reconocerlos en un contexto educativo la inclusión de políticas, sociales, culturales, religiosos, económico y su influencia en la organización de las prácticas pedagógicas. Causar problemas y plantear nuevas posibilidades para ser trilladas a la consolidación de prácticas, cuyas necesidades han revelado como indicativo establecer un hecho de que el aprendizaje se vuelve significativa y emancipadora para los sujetos que las había asignado.ABSTRACT. This article presents analysis of pedagogical practices structured by monitors / those working with high school / technical a Family Farm School (EFA), aiming to show, through the perceptions of these subjects about their practices, challenges and experienced clashes in routine of classroom spaces, whose educational project is founded on Training in Alternation with teaching / different methodological principles. Qualitative nature, the contribution of this analysis will provide to the Monitor / the resumption of reflection on their practices, without pretension to impose, but to problematize, describing increasingly the process of elaboration and re-elaboration of knowledge in the light of the educational project of alternation. Under the guidance of authors aware of the debate on Education Course and Training in Alternation, we realize that there are many factors that interfere with pedagogical where monitors/recognize them in an educational context the inclusion of political, social, cultural, religious, economic and their influence in the organization of pedagogical practices. Cause challenges and raise new possibilities to be threshed to the consolidation of practices, whose requirements have revealed as indicative establish a fact that learning becomes meaningful and emancipatory for the subjects that it was appropriated.ABSTRACT. This article presents analysis of pedagogical practices structured by monitors / those working with high school / technical a Family Farm School (EFA), aiming to show, through the perceptions of these subjects about their practices, challenges and experienced clashes in routine of classroom spaces, whose educational project is founded on Training in Alternation with teaching / different methodological principles. Qualitative nature, the contribution of this analysis will provide to the Monitor / the resumption of reflection on their practices, without pretension to impose, but to problematize, describing increasingly the process of elaboration and re-elaboration of knowledge in the light of the educational project of alternation. Under the guidance of authors aware of the debate on Education Course and Training in Alternation, we realize that there are many factors that interfere with pedagogical where monitors/recognize them in an educational context the inclusion of political, social, cultural, religious, economic and their influence in the organization of pedagogical practices. Cause challenges and raise new possibilities to be threshed to the consolidation of practices, whose requirements have revealed as indicative establish a fact that learning becomes meaningful and emancipatory for the subjects that it was appropriated

Antibacterial Action Mechanisms of Honey: Physiological Effects of Avocado, Chestnut, and Polyfloral Honey upon Staphylococcus aureus and Escherichia coli

[EN] Numerous studies have explored the antibacterial properties of different types of honey from all around the world. However, the data available describing how honey acts against bacteria are few. The aim of this study was to apply a flow cytometry (FC) protocol to examine and characterize the primary effects of three varieties of honey (avocado, chestnut and polyfloral) upon physiological status of Staphylococcus aureus and Escherichia coli cells to reveal their antibacterial action mechanisms. The effects of honey samples on membrane potential, membrane integrity, and metabolic activity were assessed using different fluorochromes, in a 180 min time course assay. Time-kill experiments were also carried out under similar conditions. Exposure of S. aureus and E. coli to the distinct honey samples resulted in physiological changes related to membrane polarization and membrane integrity. Moreover, honey induced a remarkable metabolic disruption as primary physiological effect upon S. aureus. The different honey samples induced quite similar effects on both bacteria. However, the depth of bacteria response throughout the treatment varied depending on the concentration tested and among honey varieties, probably due to compositional differences in the honeySIThis work was supported by the Consejería de Sanidad of Junta de Castilla y León; under Grant GRS 551/A/10. Patricia Combarros-Fuertes was funded by Consejería de Educación of Junta de Castilla y León and European Social Fun

#088. Prevalência de periodontite apical numa população portuguesa associada aos hábitos tabágicos

Objetivos: Determinar a prevalência de periodontite apical (PA) numa população portuguesa e perceber a influência do tabaco no status periapical. Materiais e métodos: Foram avaliados 402 processos clínicos e respetivas radiografias panorâmicas, de pacientes da Clínica Dentária do Instituto de Ciências da Saúde de Viseu da Universidade Católica Portuguesa, com pelo menos 8 dentes em boca e com mais de 18 anos, que foram tratados ou estão em tratamento desde janeiro de 2015 a março de 2016. Procedeu‐se ao registo da presença de PA associada ao hábito comportamental do tabaco, por indivíduo. A classificação quanto à severidade das lesões periapicais foi realizada segundo o índice periapical de Ørstavik (PAI). Os hábitos tabágicos foram registados em não fumadores e fumadores. Resultados: A média de idades da população estudada foi de 46,81 ± 17,5 anos, 57,2% eram do género feminino e 42,8% do género masculino, e a prevalência de indivíduos fumadores foi de 23,9% (do género masculino: 29,7% fumadores; do género feminino: 19,6% fumadores). A prevalência de PA por indivíduo foi de 53,5% sendo que, do total de indivíduos fumadores, 66,7% apresentou pelo menos uma lesão de PA (PAI > = 3). Os valores da associação entre hábitos tabágicos e prevalência de PA apresentaram‐se significativamente relacionados (p < 0,05). Conclusões: A associação entre a prevalência de PA e os hábitos tabágicos, verificada neste estudo, salienta a importância que tem sido atribuída recentemente à influência do status oral na saúde geral do indivíduo.info:eu-repo/semantics/publishedVersio