Análise de ligação do gene de resistência Zym-2 com marcadores microssatélites e reação de acessos de meloeiro ao Zucchini yellow mosaic virus (ZYMV)

As viroses causam perdas significativas na cultura do melão. Dentre essas, o vírus do mosaico amarelo da abobrinha-de-moita (Zucchini yellow mosaic virus- ZYMV) possui grande importância para a cultura e é encontrado em todos os locais de plantio de cucurbitáceas. O controle desse vírus através da resistência genética é a forma mais eficiente de manejo. O acesso PI414723 é a única fonte de resistência de meloeiro ao ZYMV. Essa resistência é oligogênica e supostamente condicionada por três genes dominantes: Zym-1, Zym-2 e Zym-3. A localização cromossômica do gene Zym-1 já foi confirmada no grupo de ligação 2, próximo ao marcador CMAG36. Entretanto, a localização de Zym-2 ainda carece de confirmação experimental, muito embora existam evidências de sua localização no grupo de ligação 10 (LGX). Sendo assim, um dos objetivos do presente trabalho foi confirmar a localização do gene Zym-2 através de análises de ligação com marcadores microssatélites (SSRs). Para tanto, foi utilizada uma população F2 derivada do cruzamento PI414723 x \'Védrantais\'. As plantas foram inoculadas mecanicamente com o isolado RN6-F, patótipo 0, duas vezes em um intervalo de 24 h. A confirmação da infecção e a quantificação dos títulos virais nas plantas F2 foram realizadas através do teste PTA-ELISA. O DNA genômico das plantas foi extraído da primeira folha verdadeira e utilizado nas reações de PCR com primers específicos para SSRs selecionados pertencentes ao LGX. Observou-se uma distribuição assimétrica de classes de absorbância e maior frequência de indivíduos F2 na classe com menor valor (0,1 a 0,2), sugerindo a existência de um gene de efeito maior. O teste chi-quadrado mostrou que todos os marcadores segregaram na frequência esperada (1:2:1), exceto o marcador CMCT134b. A ligação do Zym-2 aos marcadores foi confirmada por meio de regressão linear simples. Dos marcadores analisados, a regressão linear foi significativa para MU6549 e CMBR55, com p-valores de 0,011 e 0,0054, respectivamente. As análises de ligação mostraram que as ordens e as distâncias entre os marcadores condizem com os mapas presentes na literatura. Um segundo objetivo do estudo foi o de avaliar a reação ao ZYMV de 42 acessos de meloeiro oriundos da região Nordeste do Brasil, com o intuito de explorar novas fontes de resistência. Foram realizados dois experimentos utilizando a mesma metodologia citada anteriormente. O título viral médio entre os acessos variou de 0,123 a 0,621 no experimento 1 e de 0,019 a 0,368 no experimento 2. Alguns acessos apresentaram consistentemente baixos títulos virais, próximos aos do acesso resistente PI414723 e dos controles negativos (plantas não inoculadas da cultivar \'Védrantais\'). Portanto, estes acessos mostram-se como potenciais fontes de resistência ao vírus para o emprego em programas de melhoramento.

Characterization of adverse drug reactions with neuropsychiatric clinical manifestations reported spontaneously to the portuguese pharmacovigilance system in the greater Lisbon area

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

Infecção nosocomial por Clostridium difficile : caracterização da estirpe epidémica 027

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

Terapia génica: conceitos e aplicações na medicina dentária

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

Vacina contra a cárie dentária: perspetivas e preocupações: uma opção possível?

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

Infecções respiratórias de origem viral mais frequentes na comunidade

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

Veículos e vetores em terapia génica

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

O vírus do papiloma humano e a vacinação contra o cancro do colo do útero

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

Micropartículas de quitosana estruturadas com aerosil®: estabilidade, adsorção, encapsulação e liberação de substâncias ativas

Biodegradable microspheres used as controlled release systems are important in pharmaceutics. Chitosan biopolymer represents an attractive biomaterial alternative because of its physicochemical and biological characteristics. Chitosan microspheres are expected to become promising carrier systems for drug and vaccine delivery, especially for non-invasive ways oral, mucosal and transdermal routes. Controlling the swelling rate and swelling capacity of the hydrogel and improving the fragile nature of microspheres under acidic conditions are the key challenges that need to be overcomed in order to enable the exploration of the full pharmaceutical potential use of these microparticles. Many studies have focused on the modification of chitosan microsphere structures with cross-linkers, various polymers blends and new organic-inorganic hybrid systems in order to obtain improved properties. In this work, microspheres made of chitosan and nanosized hydrophobic silica (Aerosil R972) were produced by a method consisting of two steps. First, a preparation of a macroscopically homogeneous chitosan-hydrophobic silica dispersion was prepared followed by spray drying. FTIR spectroscopy, X-ray powder diffraction, differential scanning calorimetry, thermal gravimetric analysis, scanning electron microscopy (SEM) and high-resolution transmission electron microscopy (TEM) were used to characterize the microspheres. Also, the were conducted acid stability, moisture sorption capacity, release properties and biological assays. The chitosan-hydrophobic silica composite microspheres showed improved thermal degradation, lower water affinity, better acid stability and ability to retard rifampicin and propranolol hydrochloride (drug models) release under simulated physiological conditions. In vitro biocompatibility studies indicated low cytotoxicity and low capacity to activate cell production of the pro-inflammatory mediator nitric oxide. The results show here encourage further studies on the use of the new chitosan-hydrophobic silica composite microspheres as drug carrier systems via oral or nasal routes.

Co-circulação dos vírus dengue tipo 1, 2 e 4 nos Estados do Rio Grande do Norte e Paraíba, 2013

Dengue is a viral disease transmitted by female mosquitoes from genus Aedes, the principal urban vector is Aedes aegypti. Actually dengue has caused, in global scale, substantial morbidity and mortality. Four serotypes (antigenically distinct) are known: DENV-1, DENV-2, DENV-3 and DENV-4. The objective of this study was described the epidemiological profile dengue in the states of Rio Grande do Norte (RN) and Paraíba (PB), 2013. For that, suspected cases of dengue were studied, received for Laboratory of Molecular Biology of infectious disease and cancer (LADIC-UFRN) from different Health Units from RN and PB between January and December of 2013. The viral RNA was obtained from serum samples of patient from health units from RN and PB. It were studied 478 suspected cases of dengue , 252 (52,7%) from Rio Grande do Norte and 226 (47,3%) from Paraíba, showeds a global rate of infection global prevalence of 29,7% (142/478). The co-circulation of three serotypes was observed: DENV-1 (9,8% [14/142]), DENV-2 (3,5% [5/142]) and DENV-4 (86,7% [123/142]). People between 21-30 years old were the most affected by the disease during all the period of the study, representing 63,7% of the cases in both states. The genus most affected was female, representing 63,3% of cases in both states. Pau dos Ferros, Rio Grande do Norte, had the highest circulation of disease, with 8,2% (8/97) of cases. In Paraíba, the city most affected was João Pessoa, with (80% (36/45) of cases. The months with the biggest viral circulation in RN and PB were March and August, respectively. These results are very important to understanding the dengue viral activity in RN and PB, providing data that can guide control actions of this disease in support to local control programs

Estudo in silico das interações da protease NS3-NS2B de DENV-2 com o inibidor peptídico Bz-nKRR-H com finalidades terapêuticas

Dengue virus is an important patogen that causes Dengue desease in all world, and belongs to Flavivirus gender. The virus consists of enveloped RNA with a single strand positive sense, 11Kb genome. The RNA is translated into a polyprotein precursor, wich is cleaved into 3 structural proteins (C, prM e E) and 7 non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B e NS5). The NS3 is a multifunctional protein, that besides to promote the polyprotein precursor cleavage, also have NTPase, helicase and RTPase activity. The NS3 needs a hydrophilic segment of 40 residues from the transmembrane NS2B protein (who acts like cofator) to realize this functions. Actually, there's no vacines available on the market, and the treatment are just symptomatic. The tetrapeptide inhibitor Bz-Nle-Lys-Arg-Arg-H (Ki de 5,8-7,0 M) was showed as a potent inhibitor μ for NS3prot in Dengue virus. That is a inteligent alternative to treat the dengue desease. The present work aimed analyse the interactions of the ligand bounded to the activity site to provid a clear and depth vision of that interaction. For this purpouse, it was conducted an in silico study, by using quantum mechanical calculations based on Density Functional Theory (DFT), with Generalized Gradient approximation (GGA) to describe the effects of exchange and correlation. The interaction energy of each amino acid belonging to the binding site to the ligand was calculated the using the method of molecular fragmentation with conjugated caps (MFCC). Besides energy, we calculated the distances, types of molecular interactions and atomic groups involved. The theoretical models used were satisfactory and show a more accurate description when the dielectric constant = 20 ε and 80 was used. The results demonstrate that the interaction energy of the system reached convergence at 13.5 A. Within a radius of 13,5A the most important residues were identified. Met49, Met84 and Asp81 perform interactions of hydrogen with the ligant. The Asp79 and Asp75 residues present high energy of attraction. Arg54, Arg85 and Lys 131 perform hydrogen interactions with the ligand, however, appear in BIRD graph having high repulsion energy with the inhibitor. The data also emphasizes the importance of residue Tyr161 and the involvement of the catalytic triad composed by Asp75, His51 and Ser135

Validação do conteúdo do diagnóstico de enfermagem proteção ineficaz para pacientes submetidos à hemodiálise

Methodological study, in order to validate the content of the nursing diagnosis protection ineffective in patients undergoing hemodialysis. The research took place in two stages, namely: concept analysis and content analysis of the instrument by experts. T he first step was operationalized through an integrative review of the databaes LILACS, CINAHL, PubMed, Scopus and Cochrane, with the key words protection and hemodialysis, in October and November 2013. The sample consisted of 32 articles, which were analyz ed by a c areful reading to identify the sections that correspond ed to the defining attributes , antecedents and consequences of protection in patients undergoing hemodialysis. T he interpretation for the diagnosis of effective protection was made by transpos ing the components of the diagnosis (definition, defining characteristics and related factors) to the denial form . In the second stage, we elaborated an instrument with the components of the nursing diagnostic s studied and proceeded to the analysis conduct ed in April 2014 by 22 specialists in nephrology and in the terminology of the NANDA International, selected by means of th e Lattes Platform . We used the binomial test to assess the proportion of experts who rated each item as appropriate, considering a si gnificance level of 5%. The project was approved by the Ethics Committee of the institution responsible for the research, an opinion on num b er 387 837 and CAAE 18486413.0.0000.5537. The results show that the proposal for the nursing diagnosis of ineffectiv e protection in patients undergoing hemodialysis is: definition - the same as that presented in the NANDA International Taxonomy II, location - domain safety / protection and class injury. Related factors are: Absence of routine vaccines; Non - adherence to care related to vascular access; Non - adherence to infection control measures; Non - adherence to prescribed diet; Non - adherence to drug therapy; Presence of comorbidities; Drug abuse; Immune disorders; Extremes of age; Abnormal blood profiles; Drugs that red uce immunity; and side effects and adverse treatment - related. The defining characteristics are: Presence of invading the bloodstream; Nutritional disorders; Increase in the number of hospitalizations; Uncontrolled dry weight; Infected vascular access; Vasc ular inadequate access; Increased blood pressure; fever; Bleeding disorder; Disability immunity; fatigue; weakness; itching; and maladaptive response to stress. It follows that the identification of the defining attributes, antecedents and consequences inc reased the wealth of vocabulary, allowing the construction of theoretical and empirical definitions for a broader understanding of the concept protection. Furthermore, the study contributed to the enrichment of nursing specific body of knowledge, as well a s in the direction of nursing care for patients undergoing hemodialysis.

Spondias tuberosa Arr. (UMBU): estudo fitoquímico e avaliação do potencial anti-inflamatório

Spondias tuberosa Arruda (Anacardiaceae) is a fruitful tree popularly known as umbuzeiro, tapereba or umbu. It is a native and endemic species from Brazil, widespread in Brazilian Northeast. The species is important in folk medicine of the semi-arid Northeast, where it is mainly used to treat various inflammatory conditions, digestive problems as well as viral and bacterial infections. However, despite the common use in folk medicine, there are scarce pharmacological and phytochemicals studies that afford scientific evidence to its popular use. Therefore, this study aimed to characterize the chemical markers in S. tuberosa leaves extract, obtained by maceration ethanol:water (70:30, [v/v]), and evaluate its anti-inflammatory potential in vivo. The phytochemical profile in TLC analysis suggested the occurence of the flavonoids rutin and isoquercitrin. HPLC analysis enabled us to confirm the presence of flavonoids and also, were detected the phenolic acids, chlorogenic acid and caffeic acid. In addition was developed and validated a HPLC method to evaluate the content of the identified compounds in S. tuberosa leaves extract according to RDC 899/2003 of ANVISA and ICH Guidelines 2005. In order to evaluate the anti-inflammatory potential of S. tuberosa leaves extract, the peritonitis and paw edema models induced by carrageenan were used, administration i.p. in mice. The results highlighted the anti-inflammatory property in vivo at 125, 250 and 500 mg/kg since a decrease in leukocyte influx to the site of inflammation, diameter of the edema and the level of myeloperoxidase were observed when compared to the drug control dexamethasone (2 mg/kg, i.p. route). Taken together, the results pointed out S. tuberosa as a potential species for developing phytotherapic derivatives in according to its popular use. With regard to the characterization markers, chlorogenic acid, caffeic acid, rutin and isoquercitrin were identified and quantified in Spondias tuberosa leaves extract so they could be used in quality control analyses of the raw material and extracts of this species.

A atuação do enfermeiro no combate à dengue e a febre chikungunya: estudo comparativo nos municípios de Parnamirim e Santa Cruz/RN

The research aimed to analyze the role of nurses in the Family Health Strategy (FHS) in the control of Dengue and Chikungunya fever in the cities of Parnamirim and Santa Cruz. It is exploratory and descriptive nature of research, with a qualitative approach, which was developed with nurses of the Family Health Strategy in the municipalities of Parnamirim and Santa Cruz. All research process followed the ethical standards laid out research in Resolution No. 466/2012 of the National Health Council about research involving human beings. Data collection was performed using a semi-structured questionnaire with open and closed questions, organized into three parts: the profile of respondents, knowledge of the disease (transmission, prevention, control) and description of practices. The actions performed by nurses are punctual performed when there is growing disease of the number of cases in order to try to reduce the number of cases. The principal methodology used for them is to talk and guidelines resulting from campaigns and home visits, using leaflets and social networks. Regarding knowledge of nurses in the city of Santa Cruz on Dengue Fever and Chikungunya, is clearly some gaps. Nursing professionals know recognize a suspected case of dengue, but get confused when trying to explain to the Chikungunya fever, exhibiting the same symptoms of dengue. Although everybody had participated in a training on Chikungunya Fever and Dengue, a very limited knowledge of nurses regarding the clinical management. It is suggested that further studies are developed on the ground a few nurses have accepted to participate in the study, so that we can identify strategies, interventions, activities and nursing actions that are consistent with the reality of working in favor of coping Aedes epidemics aegypti that have positive impact on reducing the infestation rates and may be suitable and applied in other regions.

A atuação do enfermeiro no combate à dengue e a febre chikungunya: estudo comparativo nos municípios de Parnamirim e Santa Cruz/RN

The research aimed to analyze the role of nurses in the Family Health Strategy (FHS) in the control of Dengue and Chikungunya fever in the cities of Parnamirim and Santa Cruz. It is exploratory and descriptive nature of research, with a qualitative approach, which was developed with nurses of the Family Health Strategy in the municipalities of Parnamirim and Santa Cruz. All research process followed the ethical standards laid out research in Resolution No. 466/2012 of the National Health Council about research involving human beings. Data collection was performed using a semi-structured questionnaire with open and closed questions, organized into three parts: the profile of respondents, knowledge of the disease (transmission, prevention, control) and description of practices. The actions performed by nurses are punctual performed when there is growing disease of the number of cases in order to try to reduce the number of cases. The principal methodology used for them is to talk and guidelines resulting from campaigns and home visits, using leaflets and social networks. Regarding knowledge of nurses in the city of Santa Cruz on Dengue Fever and Chikungunya, is clearly some gaps. Nursing professionals know recognize a suspected case of dengue, but get confused when trying to explain to the Chikungunya fever, exhibiting the same symptoms of dengue. Although everybody had participated in a training on Chikungunya Fever and Dengue, a very limited knowledge of nurses regarding the clinical management. It is suggested that further studies are developed on the ground a few nurses have accepted to participate in the study, so that we can identify strategies, interventions, activities and nursing actions that are consistent with the reality of working in favor of coping Aedes epidemics aegypti that have positive impact on reducing the infestation rates and may be suitable and applied in other regions.