Urticária crónica, associação a tiroidite autoimune : revisão a propósito de um caso clínico

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Os efeitos do plasma não térmico à pressão atmosférica no desenvolvimento larval e anomalias morfológicas de Drosophila Melanogaster

O plasma não térmico nos cuidados de saúde é um campo emergente que tem as suas raízes na ciência de plasmas. Este tipo de investigação tem crescido rapidamente e é agora objeto de um amplo esforço de pesquisa interdisciplinar envolvendo a medicina, a biologia, a física, a química e a engenharia. Têm sido feitos vários trabalhos de modo a elucidar quais as interações das espécies produzidas pelo plasma com os sistemas vivos. É evidente que o mecanismo da interação do plasma com os sistemas vivos é complexo, em parte devido à complexidade do plasma mas principalmente devido à enorme complexidade da biologia. O principal objetivo desta dissertação foi observar os efeitos do plasma não térmico à pressão atmosférica (PNTPAs) no desenvolvimento larval e anomalias morfológicas de Drosophila melanogaster. Para o efeito, foram expostas e analisadas fenotipicamente 2.566 larvas após exposição, dos diferentes estádios (1.º, 2.º e 3.º) de desenvolvimento. Os testes foram realizados com aplicações de plasma com e sem ultra violeta, em duas linhas diferentes de Drosophila; uma linha selvagem preparada por nós e uma linha laboratorial. A análise fenotípica revelou que após exposição as larvas apresentavam alterações no fenótipo e no comportamento que não foram observadas no controlo, nomeadamente anomalias nas mudas, traqueias partidas, formação de massas melanóticas que podiam persistir até à fase adulta, excesso de gotículas lipídicas, atraso no desenvolvimento, comportamento de não alimentação e formação de pupa imatura que levava à formação de pupa precoce e morte pupal. Na fase pupal, as anomalias mais comuns estavam relacionadas com a forma do pupário (causadas pela pipação prematura), apresentando um desenvolvimento aberrante. Entre os vários fenótipos observados, o mais significativo foi o criptocefálico (alterações na eversão dos discos imaginais) levando à morte pupal. Nos adultos, as principais anomalias morfológicas foram registadas na formação e segmentação das patas, na forma e padrão das nervuras das asas e na formação do tórax. A similaridade destes resultados com trabalhos publicados relacionados com a hormona esteróide ecdisona indicam que provavelmente o PNTPA poderá ter influenciado a biossíntese e/ou a regulação da ecdisona, a principal hormona que regula o desenvolvimento e a metamorfose em Drosophila.

Caracterização dos distúrbios alimentares e os seus efeitos na cavidade oral

Dissertação para obtenção do grau de Mestre no Instituto Superior de Ciências da Saúde Egas Moniz

Ueber Cystendegeneration der Cowper'schen drüse der Frau /

Tesis doctoral Univ. Göttingen.

Avaliação funcional e estrutural de um novo peptídeo antimicrobiano do escorpião Tityus stigmurus

In Brazil, there is a high incidence of venomous animals. Among them, scorpions are highlighted by their medical importance, and for being their venom a source of several molecules with biological and pharmacological activity not yet fully understood, including several bioactive peptides. Antimicrobial peptides (AMPs) are components of the immune system in prokaryotes and eukaryotes, used in the first line of defense against microorganisms. In the present study, we characterized the first PAM previously identified through transcriptome of the venom gland of the scorpion Tityus stigmurus, named Stigmurin. The characteristics of Stigmurin were investigated by computational modeling and construction of dendrogram. In vitro tests investigated the antibacterial, antifungal, haemolytic and cytotoxic effects of crude venom and Stigmurin. In addition, the structural characteristics of Stigmurin were investigated by circular dochroism in water, 2, 2 , 2- trifluoethanol (TFE) and sodium dodecyl sulfate (SDS) and the models were refined by molecular dynamics simulations. The results showed that the selected sequence encodes a mature protein of 17 amino acid residues and the dendrogram reveals a case of convergent evolution. The crude venom showed no antimicrobial activity, however, Stigmurin exhibited a broad spectrum of antibacterial activity, with minimal inhibitory concentrations (MIC) ranging from 31.25 and 250 µg/mL for different strains, while the hemolytic activity at these concentrations was low. In cytotoxicity studies, the crude venom was unable to reduce cell viability in VERO E6 cells; in contrast, its activity in SiHa cells was significantly higher, corresponding to IC50 of 3.6 µg/mL. For Stigmurin the concentration sable to decrease cell viability of Vero E6 and SiHa cells in 50% were 275.67 µg/mL and 212.54 µg/mL, respectively. The dichroism spectra revealed the conformational flexibility, with predominating extended and β–sheet structures, as well as a remark able renaturation ability. The results suggest that Stigmurin could be considered as a potential antiinfective drug

Avaliação da concentração de vitamina A materna e de neonatos prematuros e a termo

Vitamin A is an essential nutrient for many physiological processes such as growth and development, so that their adequate nutritional state is essential during pregnancy and lactation. Lactating women and children in breastfeeding are considered risk groups for vitamin A deficiency and some factors may increase the risk of vitamin A deficiency, such as prematurity. The aim of this work was to evaluate the vitamin A concentration in preterm and term lactating women and newborns by determination of retinol in maternal serum, umbilical cord serum and breast milk collected until 72 hours postpartum. 182 mothers were recruited and divided into preterm group (GPT; n = 118) and term group (GT, n = 64). In preterm group were also analyzed transition milk (7th-15th day; n = 68) and mature milk (30th-55th day; n = 46) samples. Retinol was analyzed by high-performance liquid chromatography (HPLC). Maternal retinol concentration in serum was 48.6 ± 12.3 µg/dL in GPT and 42.8 ± 16.3 µg/dL in the GT (p <0.01). Cord serum retinol was 20.4 ± 7.4 µg/dL in GPT and 23.2 ± 7.6 µg/dL in GT (p> 0.05). Among newborns, 43% of premature and 36% of term had low levels of serum retinol in umbilical cord (<20 µg/dL). In colostrum, the retinol in preterm and term groups had an average of 100.8 ± 49.0 µg/dL and 127.5 ± 65.1 µg/dL, respectively (p <0.05). The retinol average in preterm milk increased to 112.5 ± 49.7 µg/dL in transition phase and decreased to 57.2 ± 23.4 µg/dL in mature milk, differing significantly in all stages (p <0.05). When comparing with the recommendation of vitamin A intake (400 µg/day) GT colostrum reached the recommendation for infants, but in GPT the recommendation was not achieved at any stage. Mothers of premature infants had higher serum retinol than mothers at term; however, this was not reflected in serum retinol of umbilical cord, since premature had lower concentration of retinol. Such condition can be explained due to lower maternal physiological hemodilution and placental transfer of retinol to the fetus during preterm gestation. Comparison of retinol in colostrum showed lower concentrations in GPT; however the transition phase there was a significant increase of retinol content released by the mammary gland of preterm mothers. This situation highlights a specific physiological adaptation of prematurity, likely to more contribute to formation of hepatic reserves of retinol in premature infants.

Avaliação da imagem corporal de mulheres submetidas à cirurgia oncológica mamária

Breast cancer is the second type of cancer that affects more women of reproductive age in Brazil. Surgical treatments include: conserving surgery or mastectomy. Aimed to evaluate body image of women undergoing breast cancer surgery, based on the scale Body Image After Breast Cancer Questionnaire. It is a descriptive, exploratory, transversal, with a quantitative approach. Data were collected in Norte-riograndense League Against Cancer, between the months from March to May 2015, after consideration of the Research Ethics Committee of that institution CAEE 35155714.1.0000.5293. The study population consisted of women undergoing breast onco-surgery. To calculate the sample considered the finite population, totaling 120 subjects, collected four guys the most. Data were analyzed by the software Statistical Package for Social Sciences version 20.0. The domain scores of the scale were evaluated using descriptive and inferential statistics. The surgical group mastectomy without reconstruction showed greater impairment of body image in the field "vulnerability", "Care for the body" and "transparency" in relation to other surgical types, and suggests susceptibility to cancer, body appearance and worry that disturb other. The Kruskal-Wallis test showed greater dissatisfaction with body image in the fields "body Stigma" and "transparency" to the radical neoplastic surgery over other surgical types. Dissatisfaction with body image and physical appearance was detected in this study in all six image fields present in scale, with emphasis on the "body Stigma" and "Transparency". This means that the body image disorder is formulated based on the perception of others about themselves and not by perception "self", which justifies the concern with appearance, with body and hide the consequences stemmed cancer. It is expected that the data obtained from the evaluation of body image presented in this study contribute to enable the assistance to oncocirurgiada woman breast integral, essential for the practice of Nursing.

Avaliação da imagem corporal de mulheres submetidas à cirurgia oncológica mamária

Breast cancer is the second type of cancer that affects more women of reproductive age in Brazil. Surgical treatments include: conserving surgery or mastectomy. Aimed to evaluate body image of women undergoing breast cancer surgery, based on the scale Body Image After Breast Cancer Questionnaire. It is a descriptive, exploratory, transversal, with a quantitative approach. Data were collected in Norte-riograndense League Against Cancer, between the months from March to May 2015, after consideration of the Research Ethics Committee of that institution CAEE 35155714.1.0000.5293. The study population consisted of women undergoing breast onco-surgery. To calculate the sample considered the finite population, totaling 120 subjects, collected four guys the most. Data were analyzed by the software Statistical Package for Social Sciences version 20.0. The domain scores of the scale were evaluated using descriptive and inferential statistics. The surgical group mastectomy without reconstruction showed greater impairment of body image in the field "vulnerability", "Care for the body" and "transparency" in relation to other surgical types, and suggests susceptibility to cancer, body appearance and worry that disturb other. The Kruskal-Wallis test showed greater dissatisfaction with body image in the fields "body Stigma" and "transparency" to the radical neoplastic surgery over other surgical types. Dissatisfaction with body image and physical appearance was detected in this study in all six image fields present in scale, with emphasis on the "body Stigma" and "Transparency". This means that the body image disorder is formulated based on the perception of others about themselves and not by perception "self", which justifies the concern with appearance, with body and hide the consequences stemmed cancer. It is expected that the data obtained from the evaluation of body image presented in this study contribute to enable the assistance to oncocirurgiada woman breast integral, essential for the practice of Nursing.

Qualidade do leite de búfala (Bubalus bubalis) suplementada com selênio

This study aimed to evaluate the effect of diet with selenium on the physico-chemical composition and somatic cell count of buffalo milk, and check the selenium waste in milk and minas kind fresh cheese. For the analysis of physical-chemical composition and SCC were used 2264 buffalo milk data from the Murrah, belonging to Tapuio Ltda, located in the agreste region of the state of Rio Grande do Norte, in the period 2010 to 2014. To check the amount of selenium residue in buffalo milk and fresh cheese mines were used 100 Murrah buffaloes divided into 5 lots. The batches were formed according to the production levels of animals, wherein the amount of concentrate fed varied within each lot, which received 0.08 ppm / Se / kg of concentrate. Was collected from 300 ml of tank milk from and each lot, as well as 300 gr minas cheese, from August to November 2014, with the collection of lots held only in the month of November. The samples were sent to the Pernambuco Tecnology Institute for selenium residue analysis. The effect of the seasons was contrasted in two ways, namely: Spring, Summer, Autumn and Winter season or drought and rainy season. The analyzes statistics were performed using the procedures available in the software SAS® (SAS, 2002). Selenium level provided to the animals in this study was not significant to the point of detection of their residue in milk and the cheeses. However, it influences the reduction of the CCS. The milk characteristics were not affected by selenium use only the seasons.

Qualidade do leite de búfala (Bubalus bubalis) suplementada com selênio

This study aimed to evaluate the effect of diet with selenium on the physico-chemical composition and somatic cell count of buffalo milk, and check the selenium waste in milk and minas kind fresh cheese. For the analysis of physical-chemical composition and SCC were used 2264 buffalo milk data from the Murrah, belonging to Tapuio Ltda, located in the agreste region of the state of Rio Grande do Norte, in the period 2010 to 2014. To check the amount of selenium residue in buffalo milk and fresh cheese mines were used 100 Murrah buffaloes divided into 5 lots. The batches were formed according to the production levels of animals, wherein the amount of concentrate fed varied within each lot, which received 0.08 ppm / Se / kg of concentrate. Was collected from 300 ml of tank milk from and each lot, as well as 300 gr minas cheese, from August to November 2014, with the collection of lots held only in the month of November. The samples were sent to the Pernambuco Tecnology Institute for selenium residue analysis. The effect of the seasons was contrasted in two ways, namely: Spring, Summer, Autumn and Winter season or drought and rainy season. The analyzes statistics were performed using the procedures available in the software SAS® (SAS, 2002). Selenium level provided to the animals in this study was not significant to the point of detection of their residue in milk and the cheeses. However, it influences the reduction of the CCS. The milk characteristics were not affected by selenium use only the seasons.

Imunoexpressão da proteína endonuclease apurínica/apurimidínica (APE-1) em adenomas pleomórficos e carcinomas ex-adenomas pleomórficos

Introduction: Apurinic/Apyrimidinic Endonuclease 1 (APE-1) is an essential protein for DNA base excision repair (BER) pathway and regulation of redox activities. The ability of malignant cells to recognize and repair DNA damage is an important mechanism for tumor survival, and recent studies suggest that APE-1 overexpression is related to poor prognosis in some tumors. Purpose: To analyze the immunoreactivity of APE-1 in Pleomorphic Adenomas (PA) and Carcinomas Ex Pleomorphic Adenomas (CaExPA) of salivary glands. Materials and Methods: A total of 49 tumors fixed in formalin and embedded in paraffin (33 PA and 16 CaExPA) underwent immunohistochemical study by the immunoperoxidase technique. APE-1 immunoreactivity was evaluated quantitatively by the percentage of immunopositive cells. For statistical analysis a significance level of 5% (p≤ 0.05) was adopted. Results: All cases of PA and CaExPA (n=49) were positive for APE-1, however, there was a higher expression in CaExPA, with statistically significant difference (p<0.001). There was no association between APE-1 expression and tumors of major or minor salivary gland, however, not encapsulated PA (median expression = 54.2%) showed higher expression when compared to encapsulated tumors (p=0.02). APE-1 overexpression was found mainly in cases of CaExAP with lymph node metastasis (median expression = 90.3% - p=0.002) and invasive pattern (median expression = 89.9% - p=0.003), when compared to cases without metastasis and intracapsular pattern. Conclusion: This study suggests that APE-1 is deregulated in the studied tumors. The increased expression of APE-1 is associated with the absence of complete capsule in PA and it is associated with more aggressive behavior in CaExPA.

Imunoexpressão da proteína endonuclease apurínica/apurimidínica (APE-1) em adenomas pleomórficos e carcinomas ex-adenomas pleomórficos

Introduction: Apurinic/Apyrimidinic Endonuclease 1 (APE-1) is an essential protein for DNA base excision repair (BER) pathway and regulation of redox activities. The ability of malignant cells to recognize and repair DNA damage is an important mechanism for tumor survival, and recent studies suggest that APE-1 overexpression is related to poor prognosis in some tumors. Purpose: To analyze the immunoreactivity of APE-1 in Pleomorphic Adenomas (PA) and Carcinomas Ex Pleomorphic Adenomas (CaExPA) of salivary glands. Materials and Methods: A total of 49 tumors fixed in formalin and embedded in paraffin (33 PA and 16 CaExPA) underwent immunohistochemical study by the immunoperoxidase technique. APE-1 immunoreactivity was evaluated quantitatively by the percentage of immunopositive cells. For statistical analysis a significance level of 5% (p≤ 0.05) was adopted. Results: All cases of PA and CaExPA (n=49) were positive for APE-1, however, there was a higher expression in CaExPA, with statistically significant difference (p<0.001). There was no association between APE-1 expression and tumors of major or minor salivary gland, however, not encapsulated PA (median expression = 54.2%) showed higher expression when compared to encapsulated tumors (p=0.02). APE-1 overexpression was found mainly in cases of CaExAP with lymph node metastasis (median expression = 90.3% - p=0.002) and invasive pattern (median expression = 89.9% - p=0.003), when compared to cases without metastasis and intracapsular pattern. Conclusion: This study suggests that APE-1 is deregulated in the studied tumors. The increased expression of APE-1 is associated with the absence of complete capsule in PA and it is associated with more aggressive behavior in CaExPA.

Análise da imunoexpressão de OCT4 e CD44 em neoplasias de glândulas salivares menores e maiores

Salivary gland neoplasms exhibit a wide variety of biological behavior and a high morphological diversity raises the interest in researching these lesions. The stem cells are the main source for the generation and maintenance of cell diversity, disorders in the regulation of these cells can lead to the production of altered stem cells, termed cancer stem cells capable of generate the tumor. Researches on cancer stem cells and associated proteins have been developed in some oral cancers; however, their role in salivary gland neoplasms is not well established. Thus, the aim of this study was to identify the tumor parenchyma cells exhibiting stem cell characteristics, by evaluating the immunoreactivity of OCT4 and CD44, in a number of cases of salivary gland neoplasms. The sample consisted of 20 pleomorphic adenomas, 20 mucoepidermoid carcinomas and 20 adenoid cystic carcinoma located in minor and major salivary glands. The expression of OCT4 and CD44 was evaluated by the percentage of positive cells (PP) and the intensity of expression (IE), it is realized the sum of the scores, resulting in the total score immunostaining (PIT) ranging 0-7. All studied cases showed positive expression of OCT4 and CD44 and higher values than the control groups. It was observed that for OCT4 luminal cells and non-luminal were immunostained in the case of pleomorphic adenomas and adenoid cystic carcinoma. Already the immunoreactivity of CD44 was particularly evident in the non-luminal cells of these lesions. In mucoepidermoid carcinomas for both markers, there was immunoreactivity in squamous and intermediate cells and absence of staining mucous cells. For both markers, a statistically significant higher immunostaining was verified in neoplasms located in the major salivary glands compared with lesions in the minor salivary (p<0.001). At the total sample and in the group of minor salivary glands, malignant neoplasms exhibited higher immunoreactivity for OCT4 than pleomorphic adenoma. However, there was no statistically significant difference between the lesions and between their classifications histomorphologic. Analyzing the correlation between OCT4 and CD44 immunoexpressions, a statistically significant moderate positive correlation (r = 0.444) was observed. The high expression of OCT4 and CD44 may indicate that these proteins play an important role in identifying cancer stem cells, allowing a prediction of biological behavior of salivary gland neoplasms.

Etiología y epidemiología de las mastitis humanas

La mastitis infecciosa es una patología común durante la lactancia y constituye una de las primeras causas de destete precoz. Por tanto, debería ser considerada un problema de Salud Pública relevante, ya que priva a la pareja madre-hijo de los incuestionables beneficios que la lactancia proporciona. No obstante, la mastitis humana ha sido hasta la fecha una enfermedad subestimada e infradiagnosticada, ya que habitualmente sólo se consideran mastitis los casos agudos que cursan con una sintomatología evidente y su diagnóstico microbiológico no se realiza de forma rutinaria. La etiopatogenia de la mastitis se ha relacionado con un proceso de disbiosis en la glándula mamaria, que da lugar al sobrecrecimiento de ciertas especies presentes en la leche humana. Sin embargo, la ausencia de un diagnóstico microbiológico rutinario de la leche humana hace que los estudios microbiológicos sobre esta patología sean muy escasos y que se desconozca el papel que juegan muchos agentes etiológicos. En este trabajo, el análisis microbiológico de 1.849 muestras de leche materna procedentes de mujeres con mastitis ha revelado que el género Staphylococcus constituye el primer grupo microbiano implicado en esta patología, siendo Staphylococcus epidermidis la especie aislada con mayor frecuencia (91,56% de las muestras). La especie Staphylococcus aureus se detectó en el 29,74% de los casos. Los géneros Streptocococcus y Corynebacterium constituyeron, respectivamente, el segundo (70,20%) y tercer (16,60%) grupo microbiano con mayor prevalencia en este estudio. Estos resultados ponen de manifiesto que los estafilococos coagulasa-negativos, los estreptococos del grupo viridans y las corinebacterias, habitualmente considerados microorganismos comensales y subestimados como causa de mastitis humanas, tienen un papel relevante como agentes etiológicos de esta patología. Este hecho avala que el análisis microbiológico de la leche, identificando los agentes causales a nivel de especie, es el único medio posible para obtener un diagnóstico etiológico preciso y establecer un tratamiento eficaz para la mastitis...

Avaliação da sensibilidade ao H2O2 de diferentes linhas celulares humanas: estabelecimento de modelos celulares para avaliação da capacidade antioxidante de moléculas ou extratos

Os organismos marinhos são considerados uma fonte de novos compostos bioativos com enorme potencial biotecnológico. As bactérias associadas a macroalgas têm vindo a ser estudadas devido à produção de metabolitos secundários com atividades biológicas. Neste trabalho foram isoladas e identificadas 90 bactérias associadas às macroalgas Asparagopsis armata, Bifurcaria bifurcata e Sphaerococcus coronopifolius com diferentes características fenotípicas, sendo identificadas relativamente ao seu género através da sequenciação do gene 16S RNA. A extração de compostos bioativos foi realizada com os solventes metanol e diclorometano (1:1). A capacidade antioxidante dos extratos das bactérias associadas foi avaliada através do método fluorimétrico ORAC (oxygen radical absorbent capacity), da quantificação total de polifenóis (QTP) e da capacidade de redução do radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). O efeito citotóxico do H2O2 foi testado nos modelos celulares SH-SY5Y, MCF-7 e HepG-2, representativos de células humanas neuronais, epiteliais da glândula mamária e hepáticas, respetivamente. Os extratos com maior capacidade antioxidante foram testados nos modelos celulares em condições de stress oxidativo induzido pelo H2O2. Os resultados foram revelados pelo método de 3-[4,5- dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT). O género de bactérias mais representativo identificado em associação com Asparagopsis armata, Bifurcaria Bifurcata e Sphaerococcus coronopifolius foi Vibrio sp. com 40%, 48,72% e 28,57%, respetivamente. Os géneros de bactérias menos representativos identificados em associação com Asparagopsis armata foram Bacillus sp., Cobetia sp. e Erwinia sp., com uma ocorrência de 3,33%. Por sua vez, Citricoccus sp., Cellulophaga sp., Ruegeria sp. e Staphylococcus sp. foram os géneros de bactérias menos representativos associados a Bifurcaria Bifurcata (2,56%). Os géneros menos representativos identificados em associação com Sphaerococcus coronopifolius foram Bacillus sp. e Holomonas sp. com uma ocorrência de 9,52%. O extrato da bactéria associada que apresentou maior potencial antioxidante avaliado pelos métodos de ORAC (3603,66 ± 80,14 μmol eq. Trolox/g extrato), QTP (53,854 ± 3,02 mg eq. ácido gál./g extrato) e DPPH (20,21 (14,41-28,34) μg.mL-1) foi a BB16 (Shewanella sp.), associada à alga Bifurcaria bifurcata. O efeito induzido pelo H2O2 foi bastante distinto na redução da viabilidade celular, com IC50 distintos, nas células SH-SY5Y (206,0 μM (150,4 – 282,2)), MCF-7 (450,2 μM (388,0 – 522,5)) e HepG-2 (1058,0 μM (847,3 – 1321,0)).A elevada atividade antioxidante do extrato da bactéria associada à alga Bifurcaria bifurcata (0,1mg.mL-1; BB16 – Shewanella sp.) permitiu a prevenção do efeito induzido pelo H2O2 na linha celular SH-SY5Y (IC50 - 431,7 μM (360,1 – 517,6). Em conclusão, as bactérias associadas das macroalgas Asparagopsis armata, Bifurcaria bifurcata e Sphaerococcus coronopifolius podem ser uma excelente e interessante fonte de compostos marinhos naturais com um elevado potencial antioxidante.