Relatório de estágio II de Paula Marina da Costa Silva Matos Ferreira

O presente relatório foi elaborado no âmbito da unidade curricular Estágio II do curso de Mestrado Integrado em Ciências Farmacêuticas do Instituto Universitário de Ciências da Saúde. Este relatório diz respeito à vertente de farmácia comunitária, destacando o trabalho, as experiências e as funções do farmacêutico comunitário em geral, e as Boas Práticas de Farmácia (BPF). Neste contexto, são também descritas algumas das particularidades da atividade da Farmácia União (FU), situada em Resende, na freguesia se São Martinho de Mouros (SMM)(local de estágio).

Estudo da resistência de Pneumocystis jirovecii ao cotrimoxazol em doentes com infecção VIH SIDA

Pneumocystis jirovecii é um eucariota responsável por um quadro clínico de pneumonia intersticial grave em indivíduos imunocomprometidos. Apesar do decréscimo da incidência da pneumonia por P. jirovecii (PPc) como resultado da introdução da profilaxia específica e da terapêutica antirretroviral potente (HAART, do inglês “Highly active antiretroviral therapy”), esta doença continua a ser uma das infecções oportunistas mais frequentes em doentes com infecção vírus da imunodeficiência humana/síndroma de imunodeficiência adquirida (VIH/SIDA) e com outras imunodeficiências. Os fármacos da família das sulfas, em especial o cotrimoxazol, uma combinação de sulfametoxazol e de trimetoprim, são considerados os agentes de primeira linha na profilaxia e no tratamento desta infecção. Ao longo dos últimos anos, tem sido estudada a variabilidade genética da dihidropteroato sintetase (DHPS) e da dihidrofolato reductase (DHFR), duas enzimas alvo do cotrimoxazol, de P. jirovecii com o intuito de avaliar o possível desenvolvimento de resistência a este fármaco. Por outro lado, o estudo de outros marcadores genéticos, como as regiões dos espaçadores internos transcritos (ITS, do inglês "Internal Transcribed Spacers"), em conjunto com o DHPS e o DHFR, tem sido utilizado no sentido de melhor compreender os mecanismos de desenvolvimento de resistência ao cotrimoxazol. Com o presente estudo, pretendeu-se efectuar a caracterização genética da DHPS, da DHFR e das regiões ITS de P. jirovecii, em isolados obtidos entre 1995 e 2004, em doentes imunocomprometidos. Também foi objectivo deste trabalho, estudar a relação dosgenótipos identificados nas três regiões genómicas em análise, entre si e com diversas variáveis clínicas e epidemiológicas. Entre os 403 isolados de P. jirovecii, obtidos entre 2001 e 2004, a percentagem de infecção por P. jirovecii observada foi de 64,5%. No total, 290 isolados de P. jirovecii identificados no período de 2001–2004 (260 isolados) e entre 1995–2000 (30 isolados), foram submetidos ao estudo de caracterização genética, nos três loci em estudo. Inicialmente, com o intuito de melhorar a técnica de amplificação do gene da DHPS de P. jirovecii, procedeu-se á filtração das amostras de secreções pulmonares, para remoção de DNA do hospedeiro contaminante, e foi elaborado um novo protocolo de amplificação. Apesar, de se ter verificado que o processo de microfiltração não promovia uma melhor amplificação de DNA específico de Pneumocystis, o protocolo de PCR “nested” desenhado contribuiu para a maior amplificação do gene da DHPS de P. jirovecii (P <0,001). A caracterização genética da DHPS e da DHFR, permitiu identificar o genótipo selvagem em 84,5% e 68,9%, respectivamente, dos isolados estudados. Em 15,5% dos isolados, observou-se a presença de genótipos mutantes nos codões 55 e 57 do gene da DHPS. Em relação ao gene da DHFR, 31,1% dos isolados apresentaram polimorfismos. No total, foram identificados nove locais de substituição: quatro substituições sinónimas, nas posições nucleotídicas 201, 272, 312 e 381; cinco substituições nucleotídicas não sinónimas nas posições 38, 68, 92, 154 e 200 que resultaram na alteração nos codões 13, 23, 31, 52 e 67, respectivamente. No estudo efectuado, verificou-se que as mutações do gene da DHPS, foram mais frequentes entre os isolados obtidos em 1995-2000 do que entre os isolados recolhidos entre 2001 e 2004 (P=0,056). Relativamente ao gene da DHFR e, em contraste com a maioria dos estudos publicados, observou-se uma elevada diversidade genética entre os isolados estudados.Também, nas regiões ITS de P. jirovecii, foi observada uma elevada variabilidade entre os isolados estudados. No presente trabalho, 30 genótipos diferentes foram identificados, o que evidencia o elevado grau de diversidade genética destas regiões e a sua utilidade no estudo da transmissão e da epidemiologia da PPc. No estudo efectuado, os genótipos mais frequentes foram o Eg, o Cg e o Gg. Verificou-se que o genótipo Ne foi significativamente mais frequente entre 1995–2000 (P= 0,026) enquanto que o genótipo Eg foi mais frequente entre 2001–2004 (P=0,011). Considerando a combinação dos genótipos identificados nestas três regiões genómicas, foram observados 50 haplótipos distintos em 100 isolados de P. jirovecii. A elevada diversidade intra-específica observada neste estudo multilocus demonstra o potencial que esta abordagem pode ter na diferenciação de tipos P. jirovecii distintos e a sua utilidade no estudo da transmissão e da epidemiologia da PPc. No estudo efectuado, polimorfismos nos genes da DHPS e da DHFR, presumidamente associados à exposição ao cotrimoxazol, foram detectados em doentes não expostos a este fármaco. Esta observação pode sugerir que estas sequências polimórficas possam ser adquiridos acidentalmente, por transmissão pessoa-a-pessoa ou através de uma fonte ambiental, e não somente por pressão selectiva do cotrimoxazol. Também, a identificação de genótipos idênticos, nas três regiões genómicas em estudo, em diferentes populações de doentes sugere a ocorrência de transmissão pessoa-a-pessoa. No geral, o presente trabalho contribuiu para a clarificação do papel das mutações nos genes da DHPS e da DHFR no possível desenvolvimento de resistência ao cotrimoxazol em P. jirovecii assim como para a melhor compreensão da transmissão e da epidemiologia da PPc.Pneumocystis jirovecii is an eukaryote that causes severe interstitial penumonia in immunocompromised patients. Although the widespread use of P. jirovecii pneumonia (PcP) chemoprophylaxis and highly active antiretroviral therapy (HAART), has reduced the incidence of this infection it remains an important opportunistic infection in acquired immunodeficiency syndrome (AIDS) and other immunocompromised patients. Sulpha drugs, in particular cotrimoxazole, the fixed combination of sulphamethoxazole and trimethoprim, are key agents for treatment and prophylaxis of PcP. Over the last years, the genetic variability of P. jirovecii dihydropteroate synthase (DHPS) and dihydrofolate reductase (DHFR), the enzimatic targets of cotrimoxazole, has been evaluated in order to assess the potential development of resistance to this drug. Also, the study of other genetic markers, like the internal transcribed spacers (ITS) of the nuclear rRNA, together with the DHPS and DHFR genes, has been used in order to improve the knowledge on the mechanisms involved in the development of cotrimoxazole resistance in P. jirovecii. In the present study, the main objective was to perform the genetic characterization of DHPS, DHFR and ITS regions of P. jirovecii isolates obtained between 1995 and 2004, from immunocompromised patients. Also, we intended to study the association between the genotypes identified at the three genomic regions and with several epidemiological and clinical characteristics. Overall, among the 430 P. jirovecii isolates obtained between 2001 and 2004, the frequence of P. jirovecii infection was 64,5%. Overall, 290 P. jirovecii isolates, obtained inthe time periods 1995–2000 (30) and 2001–2004 (260), were submitted to the multilocus molecular characterization study. Initially, in order to improve the amplification method of the DHPS gene, a microfiltration procedure, for the removal of human cell contamination, and a new PCR method, for amplification in specimens with low parasite load, were evaluated. Although, the microfiltration procedure did not promote the amplification of P. jirovecii specific DNA, a higher amplification rate of DHPS gene was obtained with the “nested” PCR method designed (P < 0,001). In the present work, the genetic characterization of P. jirovecii DHPS and DHFR identified the wiltype genotype in 84,5% and 68,8%, respectively, of the isolates studied. In 15,5% of the isolates, the mutant genotypes at codons 55 and 57 of the DHPS gene was observed. In the DHFR gene, 31,1% of the isolates presented polymorphic sequences with a total of nine substitution sites identified. Four synonymous substitutions, at nucleotide positions 201, 272, 312 and 381 were observed. Also, five nonsynonymous substitutions at nucleotide positions 38, 68, 92, 154 and 200, that lead to amino acid alterations at codons 13, 23, 31, 52 and 67, respectively, were detected. In this study, DHPS mutant sequences were more frequent among the isolates collected between 1995 and 2000, compared with the isolates collected in the time period 2001–2004 (P=0,056). In the DHFR gene, a high genetic diversity was observed in the isolates studied, in contrast with other reports. Also, in the P. jirovecii ITS regions, a high genetic heterogeneity was detected in the studied isolates. In the present work, 30 different genotypes were identified, which demonstrates the high genetic diversity at this locus and confirms their great utility in the study of PcP epidemiology and transmission. In this study, the most frequent genotypeswere Eg, Cg and Gg. Genotype Ne was significantly more frequent between 1995 and 2000 (P=0,026) while type Eg was more frequent in the time period 2001–2004 (P=0,011). Combining the sequences identified at these three genomic regions, 50 different haplotypes were observed among 100 P. jirovecii isolates. The great intra-specific genetic diversity observed in the multilocus study demonstrates the potential of this analysis to distinguish different types of P. jirovecii and its utility in the study of PcP epidemiology and transmission. In the present study, DHPS and DHFR polymorphisms, presumably selected by cotrimoxazole pressure, were detected in imunocompromised patients not exposed to this drug. This observation may suggest that these polymorphic sequences may be incidentally acquired, by person-to person transmission or from an environmental source, and not only by drug selective pressure. Also, the observation of common genotypes, identified at the three genomic regions studied, present in different patients’ populations, suggests the occurrence of person-to-person transmission. Overall, the present work contributed to the clarification of the role of polymorphic sequences of the DHPS and DHFR in the potential development of P. jirovecii cotrimoxazole resistance and in the improvement of PcP epidemiology and transmission

ESPANHOL GRATUITO PARA A COMUNIDADE DO PANTANAL: APROXIMANDO UNIVERSIDADE E SOCIEDADE

Trabalho apresentado no 31º SEURS - Seminário de Extensão Universitária da Região Sul, realizado em Florianópolis, SC, no período de 04 a 07 de agosto de 2013 - Universidade Federal de Santa CatarinaEste trabalho visa relatar o trabalho desenvolvido no projeto de extensão “Espanhol gratuito para a comunidade do Pantanal”. Tal projeto consiste, como próprio título anuncia, em aulas de língua espanhola para moradores da comunidade do bairro Pantanal, localizado nos arredores da UFSC, na cidade de Florianópolis. As aulas têm duração de duas horas, são realizadas nas dependências do Centro Comunitário do Pantanal e são ministradas por duas acadêmicas do curso de Letras Espanhol da UFSC. O projeto foi implementado há três meses e tem obtido resultados bem positivos, tanto em relação ao ensino/aprendizagem da língua estrangeira em questão, como em relação às expectativas dos alunos participantes do projeto. Através de uma breve pesquisa de opinião, observamos que dentre as principais motivações para buscar o curso, se destacam: a proximidade de casa; a gratuidade do curso e a curiosidade em aprender um idioma. Em relação à relevância do curso para os alunos, foram apontadas: a possibilidade de se comunicar com hispano-falantes; motivos de viagens e crescimento pessoal e/ou profissional. Sobre a satisfação dos alunos em relação às aulas, as respostas apresentam as seguintes considerações: gostando muito, muito satisfeita, adoro, muito bom, são ótimas. Dessa forma, nos parece interessante divulgar os resultados obtidos nesta ação social promovida através da universidade, como forma de levar a conhecimento público o trabalho desenvolvido em um projeto de extensão e seus efeitos para a comunidade assistida

Parents' experiences about their child's type 1 diabetes

Introduction: Type 1 Diabetes Mellitus is a chronic disease that is expanding and increasingly affects children and adolescents, resulting in changes in their lifestyle as well as in their family’s. Objective: To identify the experiences of parents regarding their child's Diabetes Mellitus. Methods: Qualitative, phenomenological study, having as participants 14 parents of children and adolescents who have diabetes and are followed up in consultations. We used the semi-structured interview and carried out content analysis. Results: Diabetes in children leads to a family restructuring and in particular to changes in diet and relationships with other children; represents a load of negative feelings and fears that lead to the demand for new parental skills and to the appreciation of health professionals and supports received. Conclusions: The results can help health professionals in the development of parental role, namely in accepting and adapting to the disease, in overcoming the difficulties and, above all, in improving the family's quality of life.ESSV, CI&DETSinfo:eu-repo/semantics/publishedVersio

Effect of paraquat and glyphosate for the control of poaceae and convolvulaceae specimens on hydric deficit

O uso de herbicidas é uma prática muito utilizada para o controle de plantas daninhas. Entretanto, sua eficácia pode ser comprometida pela menor disponibilidade de água no ambiente, influenciando na absorção, translocação e metabolismo destes pelas plantas. O presente trabalho objetivou estudar o efeito dos herbicidas paraquat (Gramoxone) e glyphosate (Roundup Original) para o controle de capim-braquiária [Urochloa decumbens (Stapf) R.D.Webster (Poaceae)] e corda-de-viola [Merremia cissoides (Lam.) Hallier f. (Convolvulaceae)], em condições de déficit hídrico. O experimento foi conduzido em casa de vegetação, em delineamento experimental inteiramente casualizado e esquema fatorial 5×3, com três repetições. Foram consideradas cinco épocas de indução de déficit hídrico (0, 1, 3, 5 e 7 dias) e três caldas de pulverização [paraquat, glyphosate e testemunha (com a aplicação de água, apenas)]. As avaliações de porcentagem de controle foram realizadas aos 2, 5, 7, 10, 14 e 21 dias após a aplicação dos tratamentos. Aos 21 dias foi determinado o teor de massa seca das plantas daninhas. Pelos resultados obtidos, observou-se que a partir de 5 dias com déficit hídrico, a eficácia dos herbicidas paraquat e glyphosate para o controle de capim-braquiária e corda-de-viola foi reduzida.The use of herbicides is very used by growers for the control of weeds. However, their effectiveness can be compromised by the reduction of water in the atmosphere, influencing its uptake, translocation and metabolism in plants. The objective of this study was to evaluate the efficacy of herbicides paraquat (Gramoxone) and glyphosate (Roundup Original) for the control of Urochloa decumbens (Stapf) R. D. Webster (Poaceae) and Merremia cissoides (Lam.) Hallier f. (Convolvulaceae) when submitted to hydric deficit. The experiment was conducted in greenhouse conditions with completely randomized design in factorial 5×3, considering three replicates. It was considered five induction conditions of hydric deficit (0, 1, 3, 5 and 7 days) and three treatments (paraquat, glyphosate and untreated check, only water). The evaluations were performed at 2, 5, 7, 10, 14 and 21 days after treatment application. At 21 days, it was also determined the dry weight of weeds. From the results obtained, it was observed that from 5 days without irrigation the herbicides reduced their effectiveness for the control of weeds

Factors that interfere with decision making in the face of resource shortages / Fatores que interferem na tomada de decisão frente à escassez de recursos

This is an exploratory study with a qualitative approach using the narrative technique strategy with 25 health professionals, aiming to describe the factors that directly interfere in decision making in the face of resource scarcity. The study identified that the management processes, the organizational culture, the professionals' lack of understanding about the scope of the Emergency Care Unit, feelings of helplessness, lack of empowerment and discomfort in professional practice interfere with decision making. It is concluded that it is necessary to intervene through professional training, improvements in the management process and health monitoring for this professional. 

Association between mir-146a and tumor necrosis factor alpha (Tnf-α) in stable coronary artery disease

Background and Objectives: Tumor necrosis factor alpha (TNF-α) is proatherogenic and associated with the risk of acute ischemic events, although the mechanisms that regulate TNF-α expression in stable coronary artery disease (SCAD) are not fully understood. We investigated whether metabolic, inflammatory, and epigenetic (microRNA (miRNA)) markers are associated with TNF-α expression in SCAD. Materials and Methods: Patients with SCAD were prospectively recruited and their metabolic and inflammatory profiles were assessed. TNF-α levels were assessed using an enzyme-linked immunosorbent assay. The relative expression of six circulating miRNAs associated with the regulation of inflammation and/or atherosclerosis was determined. Results: Of the 24 included patients with the mean age of 65 (9) years, 88% were male, and 54% were diabetic. The TNF-α levels were (median (interquartile range)) 1.0 (0.7–1.1) pg/mL. The percentage of glycosylated hemoglobin (r = 0.418, p = 0.042), serum triglyceride levels (r = 0.429, p = 0.037), and C-reactive protein levels (r = 0.407, p = 0.048) were positively correlated with TNF-α levels. Of the candidate miRNAs, miR-146a expression levels were negatively correlated with TNF-α levels (as indicated by r = 0.500, p = 0.035 for correlation between delta cycle threshold (∆Ct) miR-146a and TNF-α levels). In multivariate analysis, serum triglyceride levels and miR-146a expression levels were independently associated with TNF-α levels. miR-146 expression levels were not associated with metabolic or other inflammatory parameters and were negatively correlated with the number of coronary vessels with obstructive disease (as indicated by r = 0.556, p = 0.017 for correlation between ∆Ct miR-146a and number of diseased vessels). Conclusions: miR-146a expression levels were negatively correlated with TNF-α levels in patients with SCAD, irrespective of other metabolic or inflammatory markers, and with the severity of coronary artery disease. The results add to the knowledge on the role of miR-146a in TNF-α-based inflammation in SCAD and support future research on the potential therapeutic use of miR-146a in such a clinical scenario.publishersversionpublishe

Circulating miRNAs are associated with the systemic extent of atherosclerosis : novel observations for miR-27b and miR-146

Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).The mechanisms that regulate the systemic extent of atherosclerosis are not fully understood. We investigated whether the expression of circulating miRNAs is associated with the extent of stable atherosclerosis to a single territory or multiple territories (polyvascular) and with the severity of atherosclerosis in each territory. Ninety-four participants were prospectively recruited and divided into five age- and sex-matched groups: presenting no atherosclerosis, isolated coronary atherosclerosis, coronary and lower extremity atherosclerosis, coronary and carotid atherosclerosis, and atherosclerosis of the coronary, lower extremity, and carotid territories. The expression of six circulating miRNAs with distinct biological roles was assessed. The expression of miR-27b and miR-146 differed across groups (p < 0.05), showing a decrease in the presence of atherosclerosis, particularly in the three territories. miR-27b and miR-146 expression decreased in association with a higher severity of coronary, lower extremity, and carotid atherosclerosis. Polyvascular atherosclerosis involving the three territories was independently associated with a decreased miR-27b and miR-146 expression. Both miRNAs presented an area under the curve of ≥0.75 for predicting polyvascular atherosclerosis involving the three territories. To conclude, miR-27b and miR-146 were associated with the presence of severe polyvascular atherosclerosis and with the atherosclerosis severity in each territory. Both are potential biomarkers of severe systemic atherosclerosis.info:eu-repo/semantics/publishedVersio

Cigarette smoking, miR-27b downregulation, and peripheral artery disease : insights into the mechanisms of smoking toxicity

Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).Cigarette smoking is a risk factor for the development of peripheral artery disease (PAD), although the proatherosclerotic mediators of cigarette smoking are not entirely known. We explored whether circulating microRNAs (miRNAs) are dysregulated in cigarette smokers and associated with the presence of PAD. Ninety-four participants were recruited, including 58 individuals without and 36 with PAD, 51 never smokers, 28 prior smokers, and 15 active smokers. The relative expression of six circulating miRNAs with distinct biological roles (miR-21, miR-27b, miR-29a, miR-126, miR-146, and miR-218) was assessed. Cigarette smoking was associated with the presence of PAD in multivariate analysis. Active smokers, but not prior smokers, presented miR-27b downregulation and higher leukocyte, neutrophil, and lymphocyte counts; miR-27b expression levels were independently associated with active smoking. Considering the metabolic and/or inflammatory abnormalities induced by cigarette smoking, miR-27b was independently associated with the presence of PAD and downregulated in patients with more extensive PAD. In conclusion, the atheroprotective miR-27b was downregulated in active smokers, but not in prior smokers, and miR-27b expression was independently associated with the presence of PAD. These unreported data suggest that the proatherogenic properties of cigarette smoking are mediated by a downregulation of miR-27b, which may be attenuated by smoking cessation.info:eu-repo/semantics/publishedVersio

Analysis of electrically evoked compound action potential of the auditory nerve in children with bilateral cochlear implants

AbstractIntroductionThe cochlear implant device has the capacity to measure the electrically evoked compound action potential of the auditory nerve. The neural response telemetry is used in order to measure the electrically evoked compound action potential of the auditory nerve.ObjectiveTo analyze the electrically evoked compound action potential, through the neural response telemetry, in children with bilateral cochlear implants.MethodsThis is an analytical, prospective, longitudinal, historical cohort study. Six children, aged 1–4 years, with bilateral cochlear implant were assessed at five different intervals during their first year of cochlear implant use.ResultsThere were significant differences in follow-up time (p=0.0082) and electrode position (p=0.0019) in the T-NRT measure. There was a significant difference in the interaction between time of follow-up and electrode position (p=0.0143) when measuring the N1-P1 wave amplitude between the three electrodes at each time of follow-up.ConclusionThe electrically evoked compound action potential measurement using neural response telemetry in children with bilateral cochlear implants during the first year of follow-up was effective in demonstrating the synchronized bilateral development of the peripheral auditory pathways in the studied population