Phenotypic and functional aspects of CD8 + T cells in atopy: from populations to cells

The prevalence of allergic diseases has been increasing during the last decades. It is believed that the development of allergic diseases in susceptible individuals is dependent on T cells, especially type 2 CD4+ T cells. CD8+ T cells may also be involved in the pathophysiology of allergic diseases, albeit studies in animal models of asthma have found confounding results. With our study we attempted to clarify the phenotypic and functional properties of CD8+ T cells in allergic disease settings. The specific aims were to assess allergen-specific proliferation and cytokine synthesis of CD8+ T cells and evaluate their suppressor function on antigen-specific responses. In addition, it was also our goal to assess the presence of CD8+ T cells at the target organ and their activation status, correlating our findings with disease severity and control. In order to contextualize our cellular findings, we initially characterised our study population from an epidemiological point of view. We enrolled asthma and rhinitis patients attending the Allergy Clinic of the Cova da Beira Hospital. Asthma and rhinitis were diagnosed by a positive clinical history and specific diagnostic tests, and severity was assessed by current guidelines. We also identified the main co-morbidities in both disease settings and evaluated asthma symptoms in rhinitis patients and rhinitis symptoms in asthma patients. In the allergic groups we investigated the sensitisation profiles to aeroallergens and concluded that major allergens included grass and cereal pollen, mites and olive tree pollen. Sensitisation profiles and severity were also compared between rural and urban-based allergic patients. In this regard, we found contrasting results between asthmatic patients and patients with rhinitis. In the second part of our study we clarified the role of CD8+ T cells in allergic inflammation through basic cellular systems. We assessed proliferation capacity and cytokine synthesis in response to Dermatophagoides pteronyssinus extract, and devised a co-culture system in order to evaluate the suppressor function of CD8+ T cells. CD8+CD28+ and CD28− T cells isolated from the peripheral blood had distinct phenotypes and proliferated at different levels to common stimuli, although sharing similar cytokine production patterns. A potential suppressor activity was not found in the co-culture systems, either for CD8+CD28+ or CD28− T cells. We then studied CD8+ T cells at the target-organ, assessing activation phenotype in cells from the induced sputum of asthmatic patients. We showed that CD8+ T cells are activated at the target-organ, although we were unable to demonstrate a relationship between activation of this T cell subset and severity or control of asthma.A prevalência das doenças alérgicas tem vindo a aumentar nas últimas décadas, especialmente nos países industrializados. Estas doenças surgem em indivíduos geneticamente predispostos, sob influência de factores ambientais. A “hipótese higiénica” procurou explicar este aumento de prevalência, relacionando-o com uma diminuição do contacto com microrganismos e do número de infecções, características do modo de vida ocidental. Inicialmente, pensou-se que a redução do contacto com microrganismos impedia o desvio imune de respostas do tipo T auxiliar 2 (“T helper 2” ou Th2) para respostas do tipo Th1, que ocorre geralmente com a idade (de acordo com o paradigma Th1/Th2). A inflamação, que ocorre particularmente ao nível das vias aéreas na asma e na rinite seria resultado de uma perda de equilíbrio entre respostas do tipo Th1, importantes nos mecanismos de defesa intracelular e respostas do tipo Th2, que caracterizam a inflamação alérgica. No entanto, estudos epidemiológicos posteriores puseram esta interpretação em causa. Foi então sugerido que, por diminuição da actividade reguladora/supressora dos linfócitos T, ocorre não um desvio imune, mas sim um desenvolvimento de respostas inadequadas face a estímulos antigénicos, resultando quer em doenças alérgicas quer em doenças auto-imunes. A atopia consiste numa predisposição genética para produzir anticorpos IgE contra antigénios ambientais comuns (alergénios) e correlaciona-se com o desenvolvimento de uma ou mais doenças alérgicas. Estas caracterizam-se pela infiltração ao nível do órgão-alvo de células inflamatórias activadas, das quais se destacam os eosinófilos, os mastócitos e os linfócitos T

Dose Administration Aid Service in Community Pharmacies: Characterization and Impact Assessment

The study was conducted according to the guidelines of the Declaration of Helsinki and approved by the Ethics Commission of the University of Beira Interior (process no. CE-UBI-Pj-2021-004:ID530 and 9 February 2021).Adherence to therapies is a primary determinant of treatment success. Lack of medication adherence is often associated with medical and psychosocial issues due to complications from underlying conditions and is an enormous waste of medical resources. Dose Administration Aid Service (DAAS) can be seen as part of the solution, allowing individual medicine doses to be organized according to the dosing schedule determined by the patient’s prescriber. The most recent systematic reviews admit the possibility of a positive impact of this service. In line with this background, the study reported in this paper aimed to characterize DAAS implementation in Portugal and understand the perceptions of pharmacists and owners of community pharmacies regarding the impact of DAAS, preferred methodology types, and State contribution. The study was guided by qualitative description methodology and reported using the consolidated criteria for reporting qualitative research (COREQ) checklist. Data were collected through semi-structured interviews with 18 pharmacists and/or owners of community pharmacies. Using qualitative content analysis, we identified categories that revealed that automated weekly methodology is the preferred methodology, because of its easiness of use and lower cost of preparation. However, the investment cost was felt to be too high by the participants considering the number of potential users for implementation in practice. Participants were also unanimous in recognizing that DAAS has a very positive impact in terms of safety and medication adherence, and the majority agreed that it also helped reduce medication waste. Implications of these findings for medication adherence are discussed.info:eu-repo/semantics/publishedVersio

Haemodialysis in Diabetic Patients Modulates Inflammatory Cytokine Profile and T Cell Activation Status

Diabetic nephropathy (DN) is a common complication in patients with diabetes, and most of them need renal replacement therapy such as haemodialysis (HD). These patients have a high tendency to develop infections and exhibit anomalies in the immune system. The objective of this study was to assess the expression of activation-related markers on T cells, as well as to quantify inflammatory cytokines, before and after a single HD session in DN patients. The study involved DN patients under HD treatment who signed an informed consent form. Blood samples before and after one HD session were collected, to analyse the expression of CD25, CD69 and CD71 in T cells. We also quantified IL-12p70, IL-8, IL-10, IL-1β, TNF-α and IL-6 in serum samples using the cytometric bead array technique. After the HD session, there was an increase in the CD4/CD8 ratio due to significant alterations in both subsets. The relative percentage of CD25+ cells and CD8+ CD25+ increased significantly after the HD session, while the relative percentage of CD69 T cells decreased. There was a significant decrease in the CD25 mean fluorescence intensity values for CD4+ T, as well as in the case of CD71 in T cells after the HD session. Regarding cytokine synthesis, we found a significant increase in IL-10 and IL-6 and a decrease in IL-8 after HD session. This study showed that a HD session in DN patients affects the T cell activation status in the two major subpopulations and differentially modulates the production of inflammatory cytokines.info:eu-repo/semantics/publishedVersio

A Systematic Review

Background: Exercise leads to a robust inflammatory response mainly characterized by the mobilization of leukocytes and an increase in circulating inflammatory mediators produced by immune cells and directly from the active muscle tissue. Both positive and negative effects on immune function and susceptibility to minor illness have been observed following different training protocols. While engaging in moderate activity may enhance immune function above sedentary levels, excessive amounts of prolonged, high-intensity exercise may impair immune function. Thus, the aim of the present review was to clarify the inflammatory effects in response to different exercise intensities. Methods: Search was performed on PubMed and was completed on July 31st, 2017. The studies were eligible if they met the predefined inclusion criteria: a) observational or interventional studies, b) conducted in healthy adults (18-65 years), c) written in Portuguese, English or Spanish, d) including moderate and/or intense exercise. Eighteen articles were included. The specific components that were examined included circulating blood levels of cytokines, leukocytes, creatine kinase (CK) and C-reactive protein (CRP). The methodological quality of the included studies was assessed. Results: Most of the intervention studies showed changes in the assessed biomarkers, although these changes were not consistent. White blood cells (WBC) had an increase immediately after intensive exercise (> 64% VO2max), without alteration after moderate exercise (46-64% VO2max). The results suggested an elevation of the pro-inflammatory cytokines, namely IL-6, followed by an elevation of IL-10 that were more evident after intense exercise bouts. CRP increased both after intense and moderate exercise, with peak increases up to 28 h. CK increased only after intensive and long exercising. Conclusion: In summary, intense long exercise can lead, in general, to higher levels of inflammatory mediators, and thus might increase the risk of injury and chronic inflammation. In contrast, moderate exercise or vigorous exercise with appropriate resting periods can achieve maximum benefit.info:eu-repo/semantics/publishedVersio

T cells in sputum of asthmatic patients are activated independently of disease severity or control

Background T cells play an important role in bronchial asthma. Although airway CD4+ T cells have been extensively studied previously, there are hardly any studies relating CD8+ T cell activation and disease symptoms. Objectives The aim of this study was to analyse the association between T cell activation in induced sputum T cells and asthma severity and control; and to evaluate T cell subpopulations in the same subgroups. Methods Fifty allergic asthmatic patients were recruited and lung function testing was performed. Airway cells were obtained by sputum induction via inhalation of hypertonic saline solution. CD3, CD4, CD8, CD28, CD25 and CD69 were studied by flow cytometry in whole induced sputum and peripheral blood cells. Results Total induced sputum T cells and CD8+ T cells had a higher relative percentage of the activation markers CD25 and CD69 in comparison with peripheral blood. In sputum, the relative percentage of CD25 was higher in CD4+ T cells when compared to CD8+ T cells and the reverse was true regarding CD69. However, neither disease severity nor control were associated with the relative percentage of CD25 or CD69 expression on T cells in sputum. Conclusions Both CD4+ and CD8+ T cells are activated in the lungs and peripheral blood of asthmatic patients. However, with the possible exception of CD69+ CD8+ T lymphocytes in the sputum, there is no association between T cell activation phenotype in the target organ and disease severity or control.info:eu-repo/semantics/publishedVersio

Evaluation of allergic rhinitis and asthma control in a Portuguese community pharmacy setting

BACKGROUND: Allergic rhinitis and asthma (ARA) are frequent respiratory diseases that often coexist, causing a high social and economic impact. It is important to maintain ARA disease control to reduce the disease burden. OBJECTIVE: To assess control in patients with 1 or both pathologies through the application of validated questionnaires at community pharmacies. METHODS: A cross-sectional study was conducted in the pharmacies of the Portuguese county of Covilhã (located in the central region of Portugal with about 53,000 inhabitants). Subjects aged between 18 and 70 years who presented a prescription for an asthma and/or allergic rhinitis medication were invited to participate in the study by responding to a questionnaire that included the Control of Allergic Rhinitis and Asthma Test (CARAT). The CARAT is a validated tool to simultaneously assess the control of ARA with scores that range from 0 to 30 points, 0 meaning the worst and 30 meaning the best possible control of disease. RESULTS: Of the 224 participants, 58% were female and the median age was 48.5 years. The median CARAT score was 19 (mean = 17.8 ± 5136.4), and 87% of participants had a score < 25, indicating noncontrolled disease. Female participants, the elderly, and those with less school education responded with scores that demonstrated significantly less disease control. CONCLUSIONS: Using a simple self-assessment questionnaire, such as the CARAT, pharmacists can help identify patients with uncontrolled ARA disease, which is an important first step to change patients’ knowledge about their disease, with an ultimate goal of improving ARA outcomes.info:eu-repo/semantics/publishedVersio

Intervenciones en el control de la xerostomia en persona en situación paliativa - revisión de alcance

Introduction:Xerostomia is the subjective sensation of dry mouth; itoccurs when less saliva is secreted than the amount of water lost through evaporation and absorption from the oral mucosa. It has multiple consequences for the general and oral health ofpeople in a palliative situation, and quality of life.Objective:Toexamine and map the interventions implemented to relieve xerostomia of the person in palliative situation.Methods:A scoping review was conducted following the Joanna Briggs Institute methodology. Data search was performed in the following databases: CINAHL COMPLETE®, PUBMED®, MEDLINE COMPLETE®, SCOPUS® and SciELO®.Results: A total of 707 articles were identified, and 19 were extracted for final analysis. This scoping review considered the studies focused on people with advanced and irreversible chronic disease, in a palliative situation and at the end of life, aged 18 years or more. Interventions for the control of xerostomia are: non-pharmacological, assessment of the oral cavity, acupuncture, saliva substitutes, saliva stimulants and oral hygiene education programmes; pharmacological parasympathomimetic: pilocarpine and bethanechol chloride.Conclusion: Xerostomia is a health condition that can cause social embarrassment and chronic discomfort, with a great impact on people's quality of life. Grouping the available evidence, within this theme, can help health professionals to incorporate it into care practice, contributing to the increase in the person’s quality of life and the relief of sufferingIntrodução:Xerostomia é a sensação subjetiva de boca seca, ocorre quando menos saliva é segregada do que a quantidade de água perdida por evaporação e por absorçãoda mucosa oral. Tem múltiplas consequências para a saúde geral e oral das pessoas em situação paliativa, bem como para a qualidade de vida. Objetivo:Objetivos: Examinar e mapear as intervenções implementadas para aliviar a xerostomia da pessoa em situação paliativa.Métodos:Scoping review de acordo com as etapas previstas pelo Joanna Briggs Institute. As bases de dados utilizadas para pesquisa foram: CINAHL COMPLETE®, PUBMED®, MEDLINE COMPLETE®, SCOPUS® e SciELO®. Resultados:Identificados 707 artigos e incluídos 19 na análise final. Esta revisão, considerou os estudos focados em pessoas com doença crónica avançada e irreversível, em situação paliativa e fim de vida, com 18 anos ou mais. As intervenções para o controlo da xerostomia são do tipo: não farmacológicas, avaliação da cavidade oral, acupuntura, substitutos da saliva, estimulantes da saliva e programas educação de higiene oral; e farmacológicas, os parassimpaticomiméticos: pilocarpina e cloreto de betanecol.Conclusão:A xerostomia é uma condição de saúde que pode causar constrangimento social e desconforto crónico, com grande impacto na qualidade de vida das pessoas. Agrupar as evidências disponíveis, dentro desta temática, pode contribuir para que os profissionais de saúde as incorporem na prática assistencial, contribuindo para um aumento da qualidade de vida da pessoa e alívio do sofrimentoIntroducción:La xerostomía es la sensación subjetiva de boca seca, ocurre cuando se secreta menos saliva que la cantidad de agua perdida por evaporación y absorción de la mucosa ora. Tiene múltiples consecuencias para la salud general y bucal de las personas en situaciones paliativas, y calidad de vida.Objetivo:Examinar y mapear las intervenciones implementadas para aliviar la xerostomia de la persona en condiciones paliativas.Métodos:Revisión del alcance de acuerdo con los pasos proporcionados por el Instituto Joanna Briggs. Las bases de datos para la investigación fueron: CINAHL COMPLETE®, PUBMED®, MEDLINE COMPLETE®, SCOPUS® y SciELO®Resultados:Se identificaron 707 artículos, 19 de ellos para el análisis final. Esta revisión consideró los estudios centrados en personas con enfermedad crónica avanzada e irreversible, en situación paliativa y al final de la vida, de 18 años más. Las intervenciones para el control de la xerostomía son: no farmacológicas, evaluación de la cavidad oral, acupuntura, sustitutos de la saliva, estimulantes de la saliva y programas de educación en higiene bucal; parasimpaticomiméticos farmacológicos: pilocarpina y cloruro de betanecol.Conclusión:La xerostomía es una condición de salud que puede causar vergüenza social y malestar crónico, con un gran impacto en la calidad de vida de las personas. Agrupar la evidencia disponible, dentro de esta temática, puede ayudar a los profesionales de la salud a incorporarla a la práctica asistencial, contribuyendo al aumento de la calidad de vida de la persona y al alivio del sufrimiento.info:eu-repo/semantics/publishedVersio

A Systematic Review of the Literature

Pharmacists have a valuable role in the management of allergic rhinitis (AR) at the community pharmacy level. This role has been reported extensively in numerous papers. However, a systematic review of the available literature and a comprehensive analysis of the outcomes has not been published. This systematic review aimed to evaluate the impact of interventions developed by pharmacists on clinical AR outcomes. A thorough search was performed in three electronic databases, including studies published between January 2000 and June 2019. After the selection process, only three articles met the inclusion criteria and were further analysed. Despite the scarcity of the available studies, in all of them was clear that the pharmacist plays a pivotal role in the management of AR, significantly improving the patients' quality of life and symptom control. This systematic review also stresses the utmost importance to investigate and report practices and interventions developed by pharmacists using measurable outcomes.info:eu-repo/semantics/publishedVersio

β2-adrenergic agonists do not improve physical performance in healthy individuals

All beta-2 agonists are prohibited by the World Anti-Doping Agency (WADA, www.wada-ama.org). However, its use may be permitted in athletes who are granted a therapeutic use exemption (TUE). [...]info:eu-repo/semantics/publishedVersio

The use of DRAQ5 to monitor intracellular DNA in Escherichia coli by flow cytometry

Flow cytometry provides a rapid and high-content multiparameter analysis of individual microorganisms within a population. In the past years, several fluorescent stains were developed in order to monitor DNA content distribution and cell-cycle phases, mainly in eukaryotic cells. Recently, due to its low detection limits, several of these fluorescent stains were also applied to prokaryotic cells. In this study, the ability of a novel far-red fluorescent stain DRAQ5 in assessing intracellular DNA content distribution in Escherichia coli DH5alpha was evaluated. The results showed that a DRAQ5-labelled live E. coli suspension can be obtained by incubation of 1 x 10(6) cells/mL with 5 microM DRAQ5 in PBS buffer supplemented with EDTA (pH = 7.4) during 30 min at 37 degrees C. Flow cytometric analysis of fixed E. coli cells revealed that ethanol should be used in detriment of glutaraldehyde for DRAQ5 labelling. After the analysis of RNase and DNase digested samples, DRAQ5 was proven to be a specific DNA labelling stain. The present study demonstrates that the use of DRAQ5 as a DNA-labelling stain provides an easy assessment of intracellular DNA content and cell-cycle phases in gram-negative bacteria such as E. coli.info:eu-repo/semantics/publishedVersio