999 resultados para Fluorescência in vivo


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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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As leishmanioses estão entre as mais importantes endemias brasileiras e encontram-se entre as doenças mais negligenciadas no mundo. O arsenal terapêutico disponível é restrito, tóxico, caro e em algumas situações ineficazes, devido ao surgimento de cepas resistentes do parasito. No Brasil são registrados anualmente mais de 20 mil casos de leishmaniose tegumentar e a Leishmania braziliensis é a principal espécie causadora das formas clínicas cutânea e mucosa. Portanto tornam-se importantes estudos que conduzam ao desenvolvimento de novas alternativas terapêuticas. O objetivo do presente estudo foi avaliar a atividade da pterocarpanoquinona denominada LQB118 sobre Leishmania braziliensis in vitro e in vivo usando hamsters como modelo experimental. O efeito antiparasitário foi avaliado sobre o crescimento in vitro das formas promastigotas e sobre amastigotas intracelulares em macrófagos peritoneais de camundongos. Para avaliar o modo ação in vitro foi investigada a indução de apoptose usando marcação por TUNEL e Anexina V-FITC. O efeito sobre a modulação da ativação de macrófagos murinos foi analisada pela dosagem de óxido nítrico (reagente de Griess) e de citocinas IL-12, TNF-alfa e IL-10 (por ELISA) nos sobrenadantes de macrófagos. In vivo a atividade terapêutica da LQB 118 foi estudada em grupos de hamsters infectados com L.braziliensis na pata, tratados com a LQB118 pelas vias intralesional (100M/3x/semana) ou oral (0,5mg/5x/semana) após 7 dias de infecção durante oito semanas. A ação terapêutica foi analisada através do tamanho da lesão. A resposta imune foi avaliada durante o tratamento, pela resposta de hipersensibilidade tardia (DTH) ao antígeno total de L. braziliensis. A ação da LQB118 in vitro foi dose-dependente tanto na forma promastigota inibindo 45%, 64,7% e 99,95%, quanto nas amastigotas intracelulares 22%, 72% e 81% nas concentrações de 5M, 10M e 20M, respectivamente para ambas as formas evolutivas. A LQB118 foi capaz de induzir a externalização de fosfatidilserina em promastigotas (18,57% das células incubadas por 24 h e em 25,79% de células tratadas por 48h) e também promoveu aumento da fluorescência nas duas formas evolutivas da Leishmania quando comparadas aos controles, demonstrando a indução de fragmentação do DNA do parasito. Esta substância também foi capaz de modular a resposta dos macrófagos infectados por 24 horas aumentando de forma dose-dependente a IL-12 e NO, mantendo constante TNF-α. In vivo, na sétima semana de tratamento, observamos uma redução significativa do tamanho das lesões nos animais tratados com LQB 118 intralesional (p<0, 001) e no grupo tratado pela via oral (p<0,05) quando comparado com o controle. Estes resultados demonstram que a atividade anti-Leishmania da LQB118 é direta sobre o parasito pela indução de morte por apoptose, apresentando também uma ação moduladora da resposta dos macrófagos contribuindo para ação leishmanicida, sem alterar a morfologia da célula hospedeira e que a LQB 118 apresenta uma atividade terapêutica no modelo hamster e pode ser uma importante molécula para o desenvolvimento de um novo fármaco.

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The objective of this clinical study was to evaluate the effectiveness of the toothbrushing with and without fluoride and the daily fluoride rinse (NaF 0.05%) on produced white spot, in vivo. This was a clinical study, controlled, randomized and triple blind. Thirty patients were selected for orthodontics reasons from Orthodontics Specialization Course at the Brazilian Dental Association - Section of Rio Grande do Norte. In this study it was used 4 bicuspid upper and lower. They had orthodontic reason for extractions, in 35 days, at least. The sample had one hundred and twenty teeth that received orthodontic bands. The bands were fixed with polycarboxylate cement, and there was a space standardized between bands and one surface of teeth. The four bicuspid of each patients were randomized and nominated as A, B, C and D. These nominations determinated the sequence of the extractions and what was done in each tooth. All the patients had been submitted to the toothbrushing with or without fluoride for 35 days. After this period, the A tooth of each patient was extracted to serve as control. The others teeth (B, C and D) were extracted one by each week. The entire sample was analyzed through the clinical examination and by laser fluorescence (DIAGNOdent®) in three different times: before orthodontic bands, 28 days after fixed and then removed the bands and, the last one, 07 days after one of the three treatments (toothbrushing with or without fluoride, tooth paste with fluoride and mouth rinse with fluoride). At the beginning all groups (A, B, C and D) had the same conditions, no significant difference was found. The same situation was found in a clinical examination. The results of the DIAGNOdent® for the groups that used tooth paste without fluoride, with fluoride and mouth rinse with fluoride, after 28 days, there was no significant difference. Clinically, the white spot was formed in all teeth after 28 days. When it was compared the three treated groups, the group without fluoride in tooth paste had worst result than the others groups. But there was no significant association between the number of active and inactive white spots and the type of treatment that the teeth had received. The demineralization of the enamel surface, under the orthodontic bands, it happened in a few weeks. The exposition of the white spots in oral environmental resulted in an improvement, but it was not enough to return to the values from the base line, either for the toothbrushing and/or the use of fluorite mouth rinse. Mouth rinse and toothpaste with fluoride have showed to reduce the incidence of demineralization in the enamel, but none seems to be superior to another one in an in vivo study

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Dissertação de Mestrado, Biologia Marinha, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2016

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Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Animal, 2016.

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The use of polycaprolactone (PCL) as a biomaterial, especially in the fields of drug delivery and tissue engineering, has enjoyed significant growth. Understanding how such a device or scaffold eventually degrades in vivo is paramount as the defect site regenerates and remodels. Degradation studies of three-dimensional PCL and PCL-based composite scaffolds were conducted in vitro (in phosphate buffered saline) and in vivo (rabbit model). Results up to 6 months are reported. All samples recorded virtually no molecular weight changes after 6 months, with a maximum mass loss of only about 7% from the PCL-composite scaffolds degraded in vivo, and a minimum of 1% from PCL scaffolds. Overall, crystallinity increased slightly because of the effects of polymer recrystallization. This was also a contributory factor for the observed stiffness increment in some of the samples, while only the PCL-composite scaffold registered a decrease. Histological examination of the in vivo samples revealed good biocompatibility, with no adverse host tissue reactions up to 6 months. Preliminary results of medical-grade PCL scaffolds, which were implanted for 2 years in a critical-sized rabbit calvarial defect site, are also reported here and support our scaffold design goal for gradual and late molecular weight decreases combined with excellent long-term biocompatibility and bone regeneration. (C) 2008 Wiley Periodicals, Inc. J Biomed Mater Res 90A: 906-919, 2009

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Purpose: To determine the subbasal nerve density and tortuosity at 5 corneal locations and to investigate whether these microstructural observations correlate with corneal sensitivity. Method: Sixty eyes of 60 normal human subjects were recruited into 1 of 3 age groups, group 1: aged ,35 years, group 2: aged 35–50 years, and group 3: aged .50 years. All eyes were examined using slit-lamp biomicroscopy, noncontact corneal esthesiometry, and slit scanning in vivo confocal microscopy. Results: The mean subbasal nerve density and the mean corneal sensitivity were greatest centrally (14,731 6 6056 mm/mm2 and 0.38 6 0.21 millibars, respectively) and lowest in the nasal mid periphery (7850 6 4947 mm/mm2 and 0.49 6 0.25 millibars, respectively). The mean subbasal nerve tortuosity coefficient was greatest in the temporal mid periphery (27.3 6 6.4) and lowest in the superior mid periphery (19.3 6 14.1). There was no significant difference in mean total subbasal nerve density between age groups. However, corneal sensation (P = 0.001) and subbasal nerve tortuosity (P = 0.004) demonstrated significant differences between age groups. Subbasal nerve density only showed significant correlations with corneal sensitivity threshold in the temporal cornea and with subbasal nerve tortuosity in the inferior and nasal cornea. However, these correlations were weak. Conclusions: This study quantitatively analyzes living human corneal nerve structure and an aspect of nerve function. There is no strong correlation between subbasal nerve density and corneal sensation. This study provides useful baseline data for the normal living human cornea at central and mid-peripheral locations

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Purpose The aim of this work is to develop a more complete understanding of the in vivo histology of the human palpebral conjunctiva and tarsal plate. Methods. The upper eyelids of 11 healthy human volunteer subjects were everted, and laser scanning confocal microscopy was used to examine the various tissue layers of the palpebral conjunctiva and tarsal plate. Results The superficial and basal epithelial layers are composed of cells with gray cytoplasm and thick, light gray borders.Nuclei can not be seen. The stroma has a varied appearance; fibrous tissue is sometimes observed, interspersed with dark,amorphous lacunae, and crevases. Numerous single white or gray cells populate this tissue, and fine blood vessels are seen traversing the field. Occasional conjunctival microcysts and Langerhans cells are observed. The tarsal plate is dark and amorphous, and meibomian gland acini with convoluted borders are clearly observed. Acini are composed of an outer lining of large cuboidal cells, and differentiated secretory cells can be seen within the acini lumen. Conclusions Laser scanning confocal microscopy is capable of studying the human palpebral conjunctiva, tarsal plate, and acini of meibomian glands in vivo. The observations presented here may provide useful supplementary anatomical information relating to the morphology of this tissue.

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Describes case studies of 2 males (aged 28 and 30 yrs) in which in vivo practice was applied to needle phobia and associated vasovagal fainting. In vivo practice combines skill acquisition with an opportunity to disconfirm negative expectations. Substantial improvements in self-efficacy, anxiety, and performance were obtained. Treatment gains generalized to other settings and were maintained at 3–6 mo follow-up. In vivo practice is recommended for further research into needle phobias.

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Background: The aim of this work is to develop a more complete qualitative and quantitative understanding of the in vivo histology of the human bulbar conjunctiva. Methods: Laser scanning confocal microscopy (LSCM) was used to observe and measure morphological characteristics of the bulbar conjunctiva of 11 healthy human volunteer subjects. Results: The superficial epithelial layer of the bulbar conjunctiva is seen as a mass of small cell nuclei. Cell borders are sometimes visible. The light grey borders of basal epithelial cells are clearly visible, but nuclei can not be seen. The conjunctival stroma is comprised of a dense meshwork of white fibres, through which traverse blood vessels containing cellular elements. Orifices at the epithelial surface may represent goblet cells that have opened and expelled their contents. Goblet cells are also observed in the deeper epithelial layers, as well as conjunctival microcysts and mature forms of Langerhans cells. The bulbar conjunctiva has a mean thickness of 32.9 1.1 mm, and a superficial and basal epithelial cell density of 2212 782 and 2368 741 cells/ mm2, respectively. Overall goblet and mature Langerhans cell densities are 111 58 and 23 25 cells/mm2, respectively. Conclusions: LSCM is a powerful technique for studying the human bulbar conjunctiva in vivo and quantifying key aspects of cell morphology. The observations presented here may serve as a useful marker against which changes in conjunctival morphology due to disease, surgery, drug therapy or contact lens wear can be assessed.