Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-02-22T23:19:51Z No. of bitstreams: 1 DaiseMariaCunhaDeSousaCarvalho_DISSERT.pdf: 1677103 bytes, checksum: 02762c1d44bfc8363c0e7f941c0a3284 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-02-26T00:20:08Z (GMT) No. of bitstreams: 1 DaiseMariaCunhaDeSousaCarvalho_DISSERT.pdf: 1677103 bytes, checksum: 02762c1d44bfc8363c0e7f941c0a3284 (MD5) / Made available in DSpace on 2016-02-26T00:20:08Z (GMT). No. of bitstreams: 1 DaiseMariaCunhaDeSousaCarvalho_DISSERT.pdf: 1677103 bytes, checksum: 02762c1d44bfc8363c0e7f941c0a3284 (MD5) Previous issue date: 2015-03-31 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / A dengue ? uma doen?a infecciosa aguda, geralmente transmitida por mosquitos Aedes aegypti. O agente etiol?gico pertence ? fam?lia Flaviviridae, g?nero Flavivirus e compreende quatro sorotipos antigenicamente relacionados, por?m distintos: DENV-1, 2, 3 e 4. No Brasil, a doen?a representa um problema de sa?de p?blica nacional. O objetivo do presente estudo foi descrever os aspectos epidemiol?gicos da dengue no Estado do Rio Grande do Norte, no per?odo de 2013 a 2014. Um total de 483 amostras de sangue ou soro, no per?odo de Janeiro de 2013 a Dezembro de 2014, foi estudado pela metodologia de RT-PCR para a detec??o e tipagem viral. A infec??o foi confirmada em 36,44% (176/483) dos casos estudados. Este estudo detectou a circula??o de tr?s sorotipos do v?rus da dengue no Rio Grande do Norte, DENV-1, 2 e 4. O sorotipo predominante em 2013-2014 foi o DENV-4, representando 83,51% (81/97) e 68,35% (54/79) dos casos positivos, respectivamente. Em rela??o ? distribui??o espacial, a maioria dos casos ocorreu em Natal e Caic?, com 9,28% (9/97) e 18,99% (15/79), respectivamente. Os meses com maior circula??o viral no RN foram Mar?o, em 2013 e Maio, em 2014. O sexo feminino foi o mais acometido, representando 69,07% (67/97) em 2013 e 54,43% (43/79) em 2014. A faixa et?ria mais acometida foi a de 21-30 anos (2013) e a de 11-20 anos (2014), com 25,77% (25/97) e 20,25% (16/79) dos casos positivos, respectivamente. A an?lise filogen?tica indicou que o gen?tipo V (DENV-1) e o gen?tipo II (DENV-4) circularam no Estado. Nossos resultados fornecem informa??es sobre a din?mica dos DENV no Rio Grande do Norte, importantes para o desenvolvimento de estrat?gias de controle da doen?a. / Dengue is an acute infectious disease, usually transmitted by Aedes aegypti mosquitoes. The etiologic agents belong to the family Flaviviridae, genus Flavivirus, and occur as four antigenically related but distinct serotypes designated DENV-1, 2, 3, and 4. In Brazil, the disease represents a national public health problem. The purpose of the present study was to describe epidemiological aspects of dengue in the State of Rio Grande do Norte, from 2013 to 2014. A total of 483 blood or serum samples, collected from January 2013 to December 2014, were studied by RT-PCR for viral detection and typing. The infection was confirmed in 36.44% (176/483) of the cases studied. This study detected the circulation of three serotypes of dengue virus in Rio Grande do Norte, DENV-1, 2, and 4. The predominant serotype in 2013-2014 was the DENV-4, representing 83.51% (81/97) and 68.35% (54/79) of the positive cases, respectively. Regarding the spatial distribution, most of the cases occurred in Natal and Caic?, with 9.28% (9/97) and 18.99% (15/79), respectively. The months with the biggest viral circulation in RN were March 2013 and May 2014. The female gender was the most affected, representing 69.07% (67/97) in 2013 and 54.43% (43/79) in 2014. The most affected age groups were 21-30 years (2013) and 11-20 years (2014) with 25.77% (25/97) and 20.25% (16/79) positive cases, respectively. Phylogenetic analysis indicated that genotype V (DENV-1) and genotype II (DENV-4) circulated in the State. Our results provide information about the dynamics of DENV in the Rio Grande do Norte, important for the development of disease control strategies.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2015-12-15T17:40:30Z No. of bitstreams: 1 DiegoMachadoPereiraDaCosta_DISSERT.pdf: 1332551 bytes, checksum: 52187e5e6b08a22ed53718a69a0a533f (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2015-12-28T21:11:24Z (GMT) No. of bitstreams: 1 DiegoMachadoPereiraDaCosta_DISSERT.pdf: 1332551 bytes, checksum: 52187e5e6b08a22ed53718a69a0a533f (MD5) / Made available in DSpace on 2015-12-28T21:11:24Z (GMT). No. of bitstreams: 1 DiegoMachadoPereiraDaCosta_DISSERT.pdf: 1332551 bytes, checksum: 52187e5e6b08a22ed53718a69a0a533f (MD5) Previous issue date: 2014-03-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / A dengue ? uma doen?a viral transmitida ao homem pela picada de mosquitos do g?nero Aedes, sendo o principal vetor urbano o Aedes aegypti. Atualmente tem causado, em escala global, consider?vel mortalidade e morbidade, o que a torna um grave problema de sa?de p?blica em todo o mundo. S?o conhecidos quatro sorotipos antigenicamente distintos: v?rus dengue (DENV) tipo 1, DENV-2, DENV-3 e DENV-4. O objetivo desse estudo foi Descrever o perfil epidemiol?gico da dengue nos estados do Rio Grande do Norte (RN) e Para?ba (PB) durante o ano de 2013. Para isso, foram estudados casos suspeitos de dengue, recebidos pelo Laborat?rio de Biologia Molecular de Doen?as Infecciosas e do C?ncer (LADIC-UFRN), provenientes de diferentes unidades de sa?de dos estados do RN e PB, no per?odo compreendido entre janeiro e dezembro de 2013. O RNA viral das amostras de soro foi extra?do atrav?s do QIAmp Viral Mini Kit, sendo seguido o protocolo descrito pelo fabricante. Para detec??o e tipagem dos DENV, foi utilizada a metodologia descrita por Lanciotti (J. Clin. Microbiol,v.30, n.3, p.545-551, 1992). Foram estudados 478 casos suspeitos de dengue, sendo 252 (52,7%) do Rio Grande do Norte e 226 (47,3) da Para?ba. A infec??o por DENV foi identificada em 29,7% (142/478) dos casos. Foi observada a co-circula??o de tr?s sorotipos virais: DENV-1 (9,8% [14/142]), DENV-2 (3,5% [5/142]) e DENV-4 (86,7% [123/142]). A faixa et?ria de 21-30 foi a mais acometida pela doen?a em todo o per?odo de estudo, representando 23,9% dos casos. O g?nero mais acometido foi o feminino, representando 63,3% dos casos. O munic?pio de maior circula??o do Estado do Rio Grande do Norte foi Pau dos Ferros, com 8,2% (8/97) dos casos identificados. J? na Para?ba, o munic?pio mais afetado foi Jo?o Pessoa, com 80% (36/45) dos casos. Os meses de maior circula??o viral no RN e PB foram Mar?o e Agosto, respectivamente. Estes resultados s?o de grande import?ncia para a compreens?o da atividade dos v?rus dengue nos estados do RN e PB, fornecendo dados para nortear as a??es de controle da doen?a, em apoio aos programas locais de combate a dengue. / 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 Para?ba (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 Para?ba, 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 Para?ba, the city most affected was Jo?o 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

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-01-27T13:32:41Z No. of bitstreams: 1 FabianaLimaBezerra_TESE.pdf: 4985146 bytes, checksum: ce1c1005e6775f052912c875b1d39910 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-01-31T13:32:42Z (GMT) No. of bitstreams: 1 FabianaLimaBezerra_TESE.pdf: 4985146 bytes, checksum: ce1c1005e6775f052912c875b1d39910 (MD5) / Made available in DSpace on 2017-01-31T13:32:42Z (GMT). No. of bitstreams: 1 FabianaLimaBezerra_TESE.pdf: 4985146 bytes, checksum: ce1c1005e6775f052912c875b1d39910 (MD5) Previous issue date: 2016-06-14 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / A dengue ? a mais importante arbovirose que afeta o homem e constitui-se em um grave problema de sa?de p?blica. A doen?a ? end?mica em regi?es tropicais e subtropicais, abrangendo mais de 100 pa?ses, correspondendo ? metade da popula??o mundial. As epidemias s?o recorrentes e estima-se que os v?rus da dengue causem 390 milh?es de infec??es a cada ano, sendo uma importante causa de morbidade. Apesar desse cen?rio, atualmente, ainda n?o se disp?e de um antiviral contra a dengue. Neste contexto, diversos estudos relatam a atividade antiviral dos polissacar?deos sulfatados obtidos das algas marinhas contra v?rus envelopados, cuja a??o est? associada ? interfer?ncia nas etapas iniciais do processo de infec??o viral. Neste estudo, avaliou-se a potencial atividade antiviral de extratos brutos ricos em polissacar?deos sulfatados obtidos das algas marrons Dictyota menstrualis (EBDMens) e Dictyota mertensii (EBDM) e das algas verdes Codium isthmocladum (EBCI) e Caulerpa sertularioides (EBCS) contra o v?rus dengue 2 (DENV-2), em linhagem de c?lulas Vero, usando diferentes estrat?gias metodol?gicas (tratamento simult?neo, tratamento p?s-infec??o, pr?-tratamento da c?lula, pr?tratamento do v?rus, adsor??o, p?s-adsor??o e penetra??o). A citotoxicidade dos extratos foi analisada pelo ensaio de redu??o do MTT. O estudo da atividade antiviral foi determinado pela quantifica??o do n?mero de c?pias do RNA viral por RT-qPCR ap?s 120 h de infec??o. Nenhum extrato exibiu toxicidade para as c?lulas Vero no ensaio de redu??o do MTT na concentra??o de 100 ?g/mL. Todos os extratos exibiram atividade antiviral quando adicionados durante os primeiros 90 minutos de infec??o, demonstrando redu??o significativa no n?mero de c?pias do RNA viral ap?s 120 horas. Os extratos EBDMens e EBDM foram mais eficazes nos ensaios de pr?-tratamento da c?lula e do v?rus, e o primeiro exerceu atividade antiviral superior ao EBDM durante a adsor??o viral. Os extratos EBCI e EBCS apresentaram atividade antiviral semelhante no ensaio de pr?-tratamento da c?lula. O EBCI foi mais eficaz na redu??o da adsor??o do DENV-2 em c?lulas Vero. Por?m, o EBCS se mostrou mais eficiente nos ensaios de pr?-tratamento do v?rus e penetra??o. Dentre os extratos avaliados, o EBCS parece ser o mais efetivo no combate ao DENV-2. Estes resultados revelam o potencial desses extratos ricos em polissacar?deos sulfatados como compostos com a??o antiviral, sugerindo que eles atuam nos est?gios iniciais da infec??o por DENV-2. / Dengue is considered the most important human arboviruses, and is a serious public health problem. This disease is endemic in tropical and subtropical regions, reaching more than 100 countries, which means the half of world population, and those epidemies have been appellant. It is estimated that Dengue virus could cause 390 millions of infections each year, and is an important cause of morbidity. Although this scenario, actually doesn?t exist a dengue antivirals still. Sorts of studies have showed antivirus activity of seaweed sulfated polysaccharides against enveloped viruses, which action seems to be associated to initials steps of infection process. In this study, was evaluated the potential antiviral activity of crude extracts rich in sulfated polysaccharides obtained from the brown algae Dyctiota menstrualis (EBDMens) and Dyctiota mertensii (EBDM), and green algae Codium isthmocladium (EBCl) and Caulerpa sertularioides (EBCS), tested against Dengue virus 2 (DENV-2) in Vero cell line, by using different methodological strategies (simultaneous treatment, post-infection treatment, cell pre-treatment, virus pre-treatment, adsorption, post-adsorption and penetration). The extracts cytotoxicity was evaluated by MTT reduction assay. The study of antiviral activity was determinated by quantifying viral RNA load using RT-qPCR, after 120 hours of infection. None extract has showed toxicity against Vero Cells in MTT reduction assay with 100 ?g/mL concentration. All extracts have shown antiviral activity when added during the first 90 minutes of infection, and a significant reduction of viral RNA number of copies after 120 hours. The EBDMens and EBDM extracts were more efficient to cell and virus pretreatment assays, and the first has showed higher antiviral activity than EBDM during viral adsorption. The EBCI and EBCS extracts have presented antiviral activity very similar in cell pre-treatment assay. EBCI was more efficient in the reduction of DENV-2 adsorption in Vero cell. However, EBCS has showed more efficient in cell pre-treatment and penetration assays. Between the extracts evaluated, EBCS seems to be more effective against DENV-2. These results have showed potential action of extracts rich in sulfated polysaccharides with antivirus activity, suggesting that they act in the initials steps of infection process of DENV-2.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-09-05T21:41:06Z No. of bitstreams: 1 PolyannaSilvaMoreira_DISSERT.pdf: 1108101 bytes, checksum: 99f0f4dcef670d145f1ddf9eb582a862 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-09-18T21:33:56Z (GMT) No. of bitstreams: 1 PolyannaSilvaMoreira_DISSERT.pdf: 1108101 bytes, checksum: 99f0f4dcef670d145f1ddf9eb582a862 (MD5) / Made available in DSpace on 2017-09-18T21:33:56Z (GMT). No. of bitstreams: 1 PolyannaSilvaMoreira_DISSERT.pdf: 1108101 bytes, checksum: 99f0f4dcef670d145f1ddf9eb582a862 (MD5) Previous issue date: 2017-02-23 / A dengue ? uma arbovirose que afeta o homem, gerando uma problem?tica na sa?de p?blica do mundo, especialmente em pa?ses tropicais os quais apresentam condi??es que favorecem a dissemina??o do mosquito Aedes aegypti. Atualmente, dentre as v?rias estrat?gias para controle da doen?a, ainda n?o se tem uma vacina eficaz ou um antiviral capaz de combater essa infec??o. Assim, o objetivo do presente estudo foi avaliar a atividade antiviral de extratos obtidos da folha e frutos da planta Morinda citrifolia L. em cultura de c?lulas Vero infectadas com v?rus dengue-2 (DENV-2). Inicialmente foram obtidos os extratos brutos (hidroetan?lico) e as respectivas fra??es: hexano, clorof?rmio e acetato de etila, analisados por cromatografia. O teste de citotoxicidade do extrato bruto, res?duo aquoso e fra??es foram realizados em cultura de c?lulas Vero pelo m?todo MTT, nas concentra??es de 1000; 500; 250; 125; 62,5; 31,2 ?g/mL. O ensaio antiviral foi conduzido atrav?s das seguintes estrat?gias: c?lulas infectadas com DENV-2 (controle positivo); c?lulas mantidas com meio de cultura (controle negativo); c?lulas infectadas com DENV-2 e tratadas com o extrato ou fra??es. Ap?s cinco dias de infec??o a viabilidade celular foi avaliada pelo m?todo de MTT e o sobrenadante da cultura foi utilizado para quantifica??o viral por unidade formadora de placa (PFU). Os resultados demonstraram que a an?lise cromatogr?fica dos extratos e fra??es revelou bandas distintas e sugestivas de saponinas, terpenos e flavonoides. Tais extratos e fra??es n?o foram t?xicos para as culturas de c?lulas, com exce??o do tratamento das c?lulas com a fra??o clorof?rmio obtido da folha e as fra??es hexano e acetato de etila do fruto verde, levando a uma viabilidade pr?xima de 65%. No ensaio antiviral o controle positivo apresentou viabilidade celular em torno de 60% ap?s cinco dias de infec??o. No tratamento com os compostos obtidos da folha observou-se que ao adicionar a fra??o de acetato de etila ?s c?lulas infectadas, estas mantiveram uma viabilidade celular pr?ximo a 100% na concentra??o de 1000?g/mL e a 85% nas concentra??es de 500 e 250?g/mL. O tratamento com a fra??o hexano apresentou uma viabilidade superior ao controle positivo em todas as concentra??es. No entanto, na fra??o clorof?rmio, a viabilidade manteve-se elevada apenas nas concentra??es de 500 e 250?g/mL. O extrato bruto e a fra??o residual aquosa n?o demonstraram atividade antiviral. As c?lulas tratadas com o extrato e as diferentes fra??es obtidas dos frutos maduro e verde, apresentaram de um modo geral uma viabilidade celular pr?xima de 100% nas concentra??es de 500 e 1000 ?g/mL no fruto maduro e apenas 1000 ?g/mL no fruto verde, com exce??o das c?lulas que foram tratadas com a fra??o clorof?rmio, na qual n?o foi poss?vel observar nenhuma diferen?a significativa quando comparado ao controle positivo. Na quantifica??o viral observou-se que as c?lulas tratadas com as fra??es hexano e clorof?rmio obtidos da folha e tamb?m os extratos brutos obtidos dos frutos maduro e verde tiveram a??o antiviral, resultando na diminui??o total da carga viral. Finalmente, a partir desse estudo podemos identificar uma poss?vel atividade antiviral dos compostos obtidos de Morinda citrifolia contra o v?rus dengue. / Dengue is an arbovirosis which affects mankind, causing problems in the public health worldwide, especially in tropical countries which present conditions that favor the spread of the mosquito Aedes aegypti. Currently, among the various strategies to control the disease, there is no effective vaccine or antiviral capable of combating this infection. Thus, the aim of the present study was to evaluate the antiviral activity of leaf and fruit extracts of the plant Morinda citrifolia L. in Vero cells culture infected with dengue-2 virus (DENV-2). Initially, the crude extracts (hydroethanolic) and their fractions were obtained: hexane, chloroform and ethyl acetate, followed by chromatographic analysis. The cytotoxicity test of the crude extract, the aqueous residue and its fractions were performed in culture of Vero cells by the MTT method, at concentrations of 1000; 500; 250; 125; 62.5; 31.2 ?g/mL. The antiviral assay results were conducted through the following strategies: cells infected with DENV-2 (positive control); cells maintained with culture medium (negative control); cells infected with DENV-2 and treated with the extract or fractions. After five days of infection, cell viability was evaluated by the MTT method and culture supernatant was used for viral quantification by plaque forming unit (PFU) assay. The results showed that the chromatographic analysis of extracts and fractions present distinct bands, which could be suggestive of saponins, terpenes and flavonoids. Such extracts and fractions were not toxic to cell cultures, except for the cells treated with the chloroform fraction obtained from leaf, hexane and ethyl acetate fractions of the green fruit, leading to a near 65% viability. In the antiviral assay the positive control had 60% of cell viability after five days of infection. Among the leaf extract treatments, it was observed that infected cells treated with ethyl acetate fraction, maintained their cell viability around 100% in the concentration of 1000?g/mL and up to 85% in the concentrations of 500 and 250?g/mL. The hexane fraction treatment showed higher viability in comparison to the positive control at all concentrations. However, in the chloroform fraction, viability remained high only at concentrations of 500 and 250 ?g/mL. Crude extract and residual aqueous fraction did not show any antiviral activity. Cells treated with the extract and different fractions obtained from the mature and green fruits, presented an overall cell viability close to 100% in 500 and 1000 ?g/mL in the mature fruit and only 1000 ?g/mL in the green fruit. However, for the cells treated with the chloroform fraction, it was not possible to observe any significant difference when compared to the positive control. In the viral quantification it was observed that cells treated with hexane and chloroform fractions obtained from leaf, as well as crude extracts obtained from mature and green fruits had antiviral effect, resulting in a total viral load decrease. Finally, identified from this study, a possible antiviral activity of the compounds obtained from Morinda citrifolia against dengue virus.