944 resultados para human whole saliva
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A Diabetes Mellitus (DM) compreende um conjunto de desordens metabólicas comuns caracterizadas por hiperglicemia, que afeta diferentes órgãos do organismo. Ao longo do tempo, ocorrem danos microvasculares no glomérulo renal, retina e nervos periféricos, bem como doença macrovascular nas artérias. A composição da saliva também é afetada pela DM, com consequências na homeostasia oral. No entanto, o proteoma e o peptidoma salivar têm sido pouco explorados na DM tipo 1 e nas suas complicações crónicas. Tendo em conta o crescente interesse na saliva como fluido diagnóstico, o objetivo principal deste trabalho foi avaliar os eventos proteolíticos subjacentes à DM tipo 1 e às suas complicações microvasculares, bem como, caracterizar as alterações induzidas pela DM tipo 1 no proteoma e peptidoma salivar. A DM tipo 1 e particularmente as complicações microvasculares associadas modulam o perfil proteolítico dos fluidos biológicos, com diferenças significativas de atividade observadas na urina e saliva, atribuídas principalmente ao complexo Metaloproteinase da Matriz (MMP)-9/lipocalina associada à gelatinase de neutrófilos, aminopeptidase N, azurocidina e calicreína 1. O aumento da atividade proteolítica observado na saliva total dos diabéticos resultou no aumento da percentagem de péptidos, principalmente de um número acrescido de fragmentos de colagénio do tipo I, refletindo possivelmente um estado inflamatório crónico dos tecidos orais e periodontais. O peptidoma também corrobora uma maior suscetibilidade das proteínas salivares, especificamente, das proteínas ricas em prolina básicas (bPRP) 1, bPRP2 e proteínas ricas em prolina ácidas (aPRP) à proteólise, evidenciando a geração de fragmentos de proteínas associadas à ligação a bactérias. A análise do proteoma salivar baseada em iTRAQ mostrou uma sobre-expressão de L-plastina, fator do adenocarcinoma do pâncreas e das proteínas S100-A8 e S100-A9, enfatizando a importância do sistema imune inato na patogénese da DM tipo 1 e das complicações microvasculares associadas. A análise integrada de todas as proteínas expressas diferencialmente entre os pacientes diabéticos com ou sem complicações microvasculares e indivíduos saudáveis foi realizada com o STRING, onde se observam três conjuntos funcionalmente ligados, um compreende a interação entre o colagénio tipo I, colagénio tipo II e MMP-9, um segundo conjunto envolve a MMP-2 e o colagénio de tipo I e um terceiro conjunto composto por proteínas salivares e inflamatórias. Estes conjuntos estão associados com as vias Kegg de interação recetor-matriz extracelular, de adesão focal e migração transendotelial dos leucócitos. Por outro lado, a análise do proteoma e peptidoma salivar destacou potenciais biomarcadores para o diagnóstico e prognóstico da DM tipo 1 e das suas complicações.
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L'élastographie ultrasonore est une technique d'imagerie émergente destinée à cartographier les paramètres mécaniques des tissus biologiques, permettant ainsi d’obtenir des informations diagnostiques additionnelles pertinentes. La méthode peut ainsi être perçue comme une extension quantitative et objective de l'examen palpatoire. Diverses techniques élastographiques ont ainsi été proposées pour l'étude d'organes tels que le foie, le sein et la prostate et. L'ensemble des méthodes proposées ont en commun une succession de trois étapes bien définies: l'excitation mécanique (statique ou dynamique) de l'organe, la mesure des déplacements induits (réponse au stimulus), puis enfin, l'étape dite d'inversion, qui permet la quantification des paramètres mécaniques, via un modèle théorique préétabli. Parallèlement à la diversification des champs d'applications accessibles à l'élastographie, de nombreux efforts sont faits afin d'améliorer la précision ainsi que la robustesse des méthodes dites d'inversion. Cette thèse regroupe un ensemble de travaux théoriques et expérimentaux destinés à la validation de nouvelles méthodes d'inversion dédiées à l'étude de milieux mécaniquement inhomogènes. Ainsi, dans le contexte du diagnostic du cancer du sein, une tumeur peut être perçue comme une hétérogénéité mécanique confinée, ou inclusion, affectant la propagation d'ondes de cisaillement (stimulus dynamique). Le premier objectif de cette thèse consiste à formuler un modèle théorique capable de prédire l'interaction des ondes de cisaillement induites avec une tumeur, dont la géométrie est modélisée par une ellipse. Après validation du modèle proposé, un problème inverse est formulé permettant la quantification des paramètres viscoélastiques de l'inclusion elliptique. Dans la continuité de cet objectif, l'approche a été étendue au cas d'une hétérogénéité mécanique tridimensionnelle et sphérique avec, comme objectifs additionnels, l'applicabilité aux mesures ultrasonores par force de radiation, mais aussi à l'estimation du comportement rhéologique de l'inclusion (i.e., la variation des paramètres mécaniques avec la fréquence d'excitation). Enfin, dans le cadre de l'étude des propriétés mécaniques du sang lors de la coagulation, une approche spécifique découlant de précédents travaux réalisés au sein de notre laboratoire est proposée. Celle-ci consiste à estimer la viscoélasticité du caillot sanguin via le phénomène de résonance mécanique, ici induit par force de radiation ultrasonore. La méthode, dénommée ARFIRE (''Acoustic Radiation Force Induced Resonance Elastography'') est appliquée à l'étude de la coagulation de sang humain complet chez des sujets sains et sa reproductibilité est évaluée.
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Background: Diabetes and periodontitis produce a protein discharge that can be reflected in saliva. This study evaluates the salivary concentrations of interleukin (IL)-6, matrix metalloproteinase (MMP)-8, and osteoprotegerin (OPG) in patients with periodontitis with type 2 diabetes. Methods: Whole saliva samples were obtained from 90 subjects who were divided into four groups: healthy (control; n = 22), untreated periodontitis (UPD; n = 24), diabetes mellitus (DM; n = 20), and UPD + DM (n = 24) groups. Clinical and metabolic data were recorded. Salivary IL-6, MMP-8, and OPG concentrations were determined by a standard enzyme-linked immunosorbent assay. Results: The UPD and UPD + DM groups exhibited higher salivary IL-6 than the control and DM groups (P <0.01). The salivary MMP-8 concentrations in all diseased groups (UPD, DM, and UPD + DM) were higher than in the control group (P <0.01). The salivary OPG concentrations in the DM group were higher than in the UPD and control groups (P<0.05). In the UPD + DM group, salivary IL-6 was correlated with glycated hemoglobin (HbA1c) levels (r = 0.60; P<0.05). The regression analysis indicated that the number of remaining teeth, clinical attachment level, and IL-6 might have influenced the HbA1c levels in patients with diabetes. Conclusions: Salivary 1L-6 concentrations were elevated in patients with periodontitis with or without diabetes. Salivary MMP-8 and OPG concentrations were elevated regardless of periodontal inflammation in patients with diabetes. Therefore, periodontitis and diabetes are conditions that may interfere with protein expression and should be considered when using saliva for diagnoses. J Periodontol 2010;81:384-391.
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Background: Previous studies reported alterations in salivary flow rate and biochemical parameters of saliva in cerebral palsy (CP) individuals; however, none of these considered the type of neuromotor abnormality among CP individuals, thus it remains unclear whether the different anatomical and extended regions of the brain lesions responsible for the neurological damage in CP might include disruption of the regulatory mechanism of saliva secretion as part of the encephalopathy. The aim of this study was to evaluate salivary flow rate, pH and buffer capacity in saliva of individuals with CP, aged 3-16 years, with spastic neuromotor abnormality type and clinical patterns of involvement. Methods: Sixty-seven individuals with CP spasticity movement disorder, were divided in two groups according to age (3-8- and 9-16-years-old) and compared with 35 sibling volunteers with no neurological damage, divided in two groups according to age (3-8- and 9-16-years-old). Whole saliva was collected under slight suction and pH and buffer capacity were determined using a digital pHmeter. Buffer capacity was measured by titration using 0.01N HCL, and flow rate was calculated in ml/min. Results: In both age groups studied, whole saliva flow rate, pH and buffer capacity were significantly lower in the spastic CP group (P < 0.05). The clinical patterns of involvement did not influence the studied parameters. Conclusion: These findings show that individuals with spastic cerebral palsy present lower salivary flow rate, pH and buffer capacity that can increase the risk of oral disease in this population.
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Objectives: This study compared three methods of Streptococcus mutans and Lactobacillus spp. detection in the oral cavity: saliva swab (SS)-sample of stimulated saliva collected with swab; whole saliva (WS)-sample of 2 ml of stimulated saliva; and the dental plaque method (DP)-plaque sample of all dental surfaces.Methods: Thirty children were included in this study. In the first 15 children, the SS and WS methods were carried out before the dental plaque collection, and in the following 15, the sequence was inverted to evaluate possible interference of the methods sequence. The samples were diluted and inoculated in SB20 and Rogosa agar, respectively for S. mutans and Lactobacillus spp., at 37 degrees C for 48 h.Results: the results (cfu/mL) of S. mutans were analysed by the statistical Friedman's test. The levels of Lactobacillus spp. were analysed by descriptive statistics due to the high proportion of zero counts in the culture. In the first sequence of methods, the number of S. mutans counted for the SS method was inferior to DP and WS (P < 0.05), and the results for the WS and DP methods were similar. The detection of Lactobacillus spp. was observed just by the WS (100 %) and SS (14.3 %) methods. However, in the second experimental set the number of S. mutans detected by the DP method was similar to those of the SS and WS, however, the WS method showed higher values than SS (P < 0.05). A greater number of Lactobacillus spp. was detected by the WS method (100 %), followed by SS (55.5 %) and DP (33.3 %).Conclusions: the dental plaque collection and the sample of stimulated whole saliva presented similar results in the S. mutans count. The most suitable method to detect the Lactobacillus spp. level in the oral cavity is the stimulated whole saliva method. (c) 2004 Elsevier Ltd. All rights reserved.
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Background. Mucocele is a lesion that involves the salivary glands and respective current ducts caused mainly by traumas in the affected area. Two different histological forms can be found: extravasation phenomenon and mucus-retention cyst where the former is the most frequently observed involving minor salivary glands such as the glands present in the anterior portion of the ventral surface of the tongue (glands of Blandin-Nuhn). Case Report. This report describes a large lesion involving the ventral surface of the tongue that was definitively diagnosed by histological examination as extravasation mucocele. Conclusion. Important concepts are reviewed to help clinicians correctly diagnose and treat this pathology. © 2006 The Authors.
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Purpose: This study aims to review anatomical, clinical, and pathological concepts as well as to discuss the most adequate therapeutic approach to the mucoceles of the glands of Blandin-Nuhn. Discussion: The glands of Blandin-Nuhn are localized in the ventral part of the tongue, next to the apex in the lingual median plane. Development of a mucocele in this site is rarely seen; besides, as the glands of Blandin-Nuhn are not encapsulated and are directly overlapped to the muscle tissues, their manipulation tends to be different from the other oral mucoceles. Conclusion: As Blandin-Nuhn mucoceles are uncommon and their clinical appearance could be similar to other lesions, it is important that health professionals know their clinical and histopathological features to avoid having them misdiagnosed. © 2010 Springer-Verlag.
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This study aimed to determine whether dental calculus formation is really higher among patients with chronic kidney disease undergoing hemodialysis than among controls. Furthermore, the study evaluated correlations between dental calculus formation and dental plaque, variables that are related to renal disease and/or saliva composition. The Renal Group was composed of 30 patients undergoing hemodialysis, whereas the Healthy Group had 30 clinically healthy patients. Stimulated whole saliva and parotid saliva were collected. Salivary flow rate and calcium and phosphate concentrations were determined. In the Renal Group the saliva collection was carried out before and after a hemodialysis session. Patients from both groups received intraoral exams, oral hygiene instructions, and dental scaling. Three months later, the dental calculus was measured by the Volpe-Manhold method to determine the rate of dental calculus formation. The Renal Group presented a higher rate of dental calculus formation (p < 0.01). Correlation was observed between rate of dental calculus formation and whole saliva flow rate in the Renal Group after a hemodialysis session (r = 0.44, p < 0.05). The presence of dental calculus was associated with phosphate concentration in whole saliva from the Renal Group (p < 0.05). In conclusion, patients undergoing hemodialysis presented accelerated dental calculus formation, probably due to salivary variables.
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Aim: The primary aim of this longitudinal study was to evaluate additional effects of 4-week chlorhexidine digluconate (CHX) gel treatments to control Aggregatibacter actinomycetemcomitans counts in children after professional dental prophylaxis. Porphyromonas gingivalis and Streptococcus mutans counts were also determined to evaluate the secondary effects of anti-plaque treatments on microbial shifts. Methods: Twenty-six children with A. actinomycetemcomitans counts >4 log10/ mL of saliva and/or Quigley-Hein plaque index >3.0 were enrolled in this study. Patients were randomly assigned to groups GI (placebo gel), GII (0.5% CHX gel), GIII (1% CHX gel), and GIV (2% CHX gel). Four sessions of treatment were performed during 4 weeks after a session of professional dental prophylaxis. Real-Time polymerase chain reaction (PCR) was used to determine viable microorganism counts in non-stimulated whole saliva samples collected at baseline, one week, one month and three months after interruption of treatments. Results: A reduction of all bacterial counts was detected after the 3-month follow-up in all groups. Lower counts of P. gingivalis were achieved from 1 week on after treatments. The 2% CHX concentration seemed to contribute to lower A. actinomycetemcomitans levels and increase S. mutans levels. Conclusions: Professional dental prophylaxis was effective to control salivary levels of A. actinomycetemcomitans, P. gingivalis and S. mutans. Additional antimicrobial effects, however, were not observed by the combination of professional dental prophylaxis and 4-week chlorhexidine gel treatments.
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Nanopartikuläre Wirkstofftransportsysteme besitzen ein großes Potential für therapeutische Anwendungen. In der vorliegenden Arbeit wurden verschiedene grundlegende Aspekte, die für das erweiterte biologische Verständnis und die Entwicklung weiterer zielgerichteter Strategien zur Pharmakotherapie mit Nanopartikeln und –kapseln notwendig sind, näher untersucht. Experimente zur zellulären Aufnahmefähigkeit (in vitro und ex vivo) wurden mit verschiedenen Nanopartikeln und –kapseln aus diversen Monomeren und biokompatiblen Makromolekülen in immortalisierten Zellkulturlinien, humanen mesenchymalen Stammzellen und Leukozyten durchgeführt und durchflusszytometrisch sowie mittels konfokaler Laser-Raster-Mikroskopie analysiert. Die Einflüsse der Oberflächenfunktionalisierungen der nanopartikulären Systeme, deren toxikologische Effekte sowie der Einfluss von adsorbiertem bovinem Serumalbumin auf funktionalisierten Polystyrol-Nanopartikeln wurden in Bezug auf die zelluläre Aufnahme untersucht.Um die multiplen Wechselwirkungen der Nanopartikel mit Bestandteilen des humanen peripheren Vollblutes zu untersuchen, wurde erfolgreich ein durchflusszytometrisches Analyseverfahren in antikoaguliertem peripherem Vollblut (ex vivo) entwickelt. Es konnte nachgewiesen werden, dass der Einfluss von Calcium-komplexierenden Antikoagulanzien zu einer Verringerung und nicht Li-Heparin zu einer Verstärkung der zellulären Aufnahme von funktionalisierten Polystyrol-Nanopartikeln in diversen Leukozyten führt.Für Folsäure-gekoppelte Hydroxyethylstärke-Nanokapseln (Synthese Frau Dr. Grit Baier) konnte ein größenabhängiger selektiver, Folatrezeptor α vermittelter, zellulärer Aufnahmeweg in HeLa-Zellen nachgewiesen werden.Hydrolysierbare, nicht zytotoxische Polyester-Nanopartikel aus Poly(5,6-Benzo-2-methylen-1,3-dioxepan) (Synthese Herr Dr. Jörg Max Siebert) mit eingebettetem Paclitaxel zeigten in HeLa-Zellen eine vergleichbare pharmakologische Wirkung wie kommerziell erhältliche Paclitaxel-Formulierungen.Die in dieser Arbeit eingesetzten Nanopartikel und Nanokapseln besitzen ein vielfältiges Potential als Wirkstofftransportsysteme. Es zeigte sich, dass Unterschiede bei der Größe, der Größenverteilung, des Polymers sowie der Oberflächenfunktionalisierung der Nanopartikel bedeutende Unterschiede der Zellaufnahme in diversen Zellkulturlinien (in vitro) und Leukozyten in peripherem Vollblut (ex vivo) zur Folge haben.
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This dissertation describes the synthesis of surface attached hydrogel biomaterials, characterization of their properties, evaluation of structuring concepts and the investigation of these materials in the isolation of DNA from human whole blood. Photosensitive hydrogel precursor materials on the basis of hydroxyethylmethacrylate (HEMA) were synthesized by free radical polymerization. In order to obtain surface bound hydrogel films, the precursors were deposited on a suitable substrate and subsequently irradatiated with UV - light to accomplish the formation of crosslinks in the film and create surface attachment. The composition of the polymerization precursor materials was determined by comprehensive NMR and GPC studies, revealing the copolymerizationrnbehaviour of the used monomers - HEMA derivatives and the photocrosslinkerrnMABP - and their respective distribution in the hydrogel precursors. The degree of crosslinking of the hydrogels was characterized with UV/vis spectroscopy. Stress-strain measurements were conducted in order to investigate the mechanical properties of the biomaterials. Moreover, the swelling process and biomolecule adsorption properties of the hydrogels were investigated with SPR/OW spectroscopy. For this, the deposition and binding of the hydrogels on gold or SiO2 surfaces was facilitated with photocrosslinkable adhesion promotors. The produced hydrogels were mechanically rigid and stablernunder the conditions of PCR and blood lysis. Furthermore, strategies towards the increase of hydrogel surface structure and porosity with porosigens, 2D laser interference lithography and photocleavable blockcopolymers were investigated. At last, a combinatorial strategy was used for the determination of the usefulness of hydrogels for the isolation from DNA from blood. A series of functionalized hydrogel precursors were synthesized, transferred to the surface inside a PCR tube and subsequently screened in regard to DNA adsorption properties with Taqman quantitative PCR. This approach yielded a promising candidate for a functional PCR tube coating that would allow the entire DNA isolation procedure being carried out in a single reaction container.rnThereforce, the practical application of such macromolecular architectures can be envisioned to improve industrial DNA diagnostic processes.
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The round window placement of a floating mass transducer (FMT) is a new approach for coupling an implantable hearing system to the cochlea. We evaluated the vibration transfer to the cochlear fluids of an FMT placed at the round window (rwFMT) with special attention to the role of bone conduction. A posterior tympanotomy was performed on eleven ears of seven human whole head specimens. Several rwFMT setups were examined using laser Doppler vibrometry measurements at the stapes and the promontory. In three ears, the vibrations of a bone anchored hearing aid (BAHA) and an FMT fixed to the promontory (pFMT) were compared to explore the role of bone conduction. Vibration transmission to the measuring point at the stapes was best when the rwFMT was perpendicularly placed in the round window and underlayed with connective tissue. Fixation of the rwFMT to the round window exhibited significantly lower vibration transmission. Although measurable, bone conduction from the pFMT was much lower than that of the BAHA. Our results suggest that the rwFMT does not act as a small bone anchored hearing aid, but instead, acts as a direct vibratory stimulator of the round window membrane.
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The rhizome of ginger (Zingiber officinale) is employed in Asian traditional medicine to treat mild forms of rheumatoid arthritis and fever. We have profiled ginger constituents for robust effects on proinflammatory signaling and cytokine expression in a validated assay using human whole blood. Independent of the stimulus used (LPS, PMA, anti-CD28 Ab, anti-CD3 Ab, and thapsigargin), ginger constituents potently and specifically inhibited IL-1β expression in monocytes/macrophages. Both the calcium-independent phospholipase A(2) (iPLA(2))-triggered maturation and the cytosolic phospholipase A(2) (cPLA(2))-dependent secretion of IL-1β from isolated human monocytes were inhibited. In a fluorescence-coupled PLA(2) assay, most major ginger phenylpropanoids directly inhibited i/cPLA(2) from U937 macrophages, but not hog pancreas secretory phospholipase A(2). The effects of the ginger constituents were additive and the potency comparable to the mechanism-based inhibitor bromoenol lactone for iPLA(2) and methyl arachidonyl fluorophosphonate for cPLA(2), with 10-gingerol/-shogaol being most effective. Furthermore, a ginger extract (2 μg/ml) and 10-shogaol (2 μM) potently inhibited the release of PGE(2) and thromboxane B2 (>50%) and partially also leukotriene B(4) in LPS-stimulated macrophages. Intriguingly, the total cellular arachidonic acid was increased 2- to 3-fold in U937 cells under all experimental conditions. Our data show that the concurrent inhibition of iPLA(2) and prostanoid production causes an accumulation of free intracellular arachidonic acid by disrupting the phospholipid deacylation-reacylation cycle. The inhibition of i/cPLA(2), the resulting attenuation of IL-1β secretion, and the simultaneous inhibition of prostanoid production by common ginger phenylpropanoids uncover a new anti-inflammatory molecular mechanism of dietary ginger that may be exploited therapeutically.
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To test the hypothesis whether microbiota in oral biofilm is linked with obesity in adolescents we designed this cross-sectional study. Obese adolescents (n = 29) with a mean age of 14.7 years and normal weight subjects (n = 58) matched by age and gender were examined with respect to visible plaque index (VPI%) and gingival inflammation (bleeding on probing (BOP%)). Stimulated saliva was collected. They answered a questionnaire concerning medical history, medication, oral hygiene habits, smoking habits, and sociodemographic background. Microbiological samples taken from the gingival crevice was analyzed by checkerboard DNA-DNA hybridization technique. The sum of bacterial cells in subgingival biofilm was significantly associated with obesity (P < 0.001). The link between sum of bacterial cells and obesity was not confounded by any of the studied variables (chronic disease, medication, VPI%, BOP%, flow rate of whole saliva, or meal frequency). Totally 23 bacterial species were present in approximately threefold higher amounts, on average, in obese subjects compared with normal weight controls. Of the Proteobacteria phylum, Campylobacter rectus and Neisseria mucosa were present in sixfold higher amounts among obese subjects. The association between obesity and sum of bacterial cells in oral subgingival biofilm indicates a possible link between oral microbiota and obesity in adolescents.