Introduktion och syfte: Kariesutveckling influeras av faktorer hos både värd och bakterier. Men när olika individer exponeras för samma nivåer av externa riskfaktorer, är en del individer mer mottagliga för karies jämfört med andra. En förklaring skulle kunna vara olika glykosylering av glykoprotein i saliven. I denna pilotstudie undersökte vi skillnaden i nivåer av monosackariderna sialinsyra, fukos och galaktos hos personer som aldrig haft karies och personer som har/har haft karies. Syftet var även att undersöka om plackenzym kan vara en modifierare av nämnda glykoprotein.Material och metod: Två grupper, med 10 individer i varje, inkluderades i studien. Ena gruppen hade DMFT = 0 och den andra DMFT ≥ 1. Saliv och plack samlades och innehållet av bundna monosackarider (sialinsyra, fukos och galaktos) samt glykosidaser (sialidas, β-fukosidas, β-galaktosidas, α-glukosidas och N-acetylglukosaminidas) analyserades med en fluorometer. Även salivflödet kalkylerades.Resultat: Innehållet av både sialinsyra och galaktos var signifikant högre i gruppen med DMFT = 0, medan innehållet av fukos inte skilde sig åt signifikant mellan grupperna. Ingen signifikant skillnad kunde ses mellan de två grupperna avseende enzymaktivitet och salivflöde.Konklusion: Högre nivåer av bunden sialinsyra och galaktos fanns hos gruppen med DMFT = 0. Resultaten indikerar att dessa monosackarider kan vara en möjlig markör för oral hälsa. Större longitudinella studier behövs för att verifiera sambandet. / Introduction and aim: Caries development is affected by factors within bacteria and host. But when different individuals are exposed to same levels of external risk factors, some individuals are still more susceptible to caries. One explanation could be different glycosylation of salivary glycoproteins. In this pilot study, we investigated the difference in levels of the monosaccharides sialic acid, fucose and galactose between individuals with or without previous caries experience. We also aimed to investigate if plaque glycosidases could be a modifier of these glycoproteins.Material and method: Two groups, with 10 subject in each, were included in this study. One group had DMFT = 0 and the other DMFT ≥ 1. Saliva and plaque were collected and content of bound monosaccharides (sialic acid, fucose and galactose) and glycosidases (sialidase, α-fucosidase, β-galactosidase, α-glucosidase and N-acetylglucosaminidase) were detected using absorbance and fluoroscens respectively. Salivary flow rate was also measured.Results: Content of both sialic acid and galactose were significantly higher in the group with DMFT = 0, while the content of fucose did not differ significantly between the groups. No significant differences could be seen between the two groups (DMFT = 0 and DMFT ≥ 1) regarding any of the investigated glycosidases and salivary flow rate. Conclusion: Higher levels of bound sialic acid and galactose were found in the group with DMFT = 0 and the results indicate that these monosaccharides could be a possible marker for oral health. Larger longitudinal studies are needed to verify this correlation.

Tesis por compendio / [ES] Se ha estimado que obtenemos más de las tres cuartas partes del nitrato que ingerimos de la fruta y la verdura. Los vegetales ricos en nitratos incluyen verduras de hoja verde y ciertos tubérculos (p. ej., remolachas y rábanos). Las glándulas salivales concentran activamente el nitrato plasmático, lo que da lugar a concentraciones elevadas de nitrato en la saliva (5 a 8 mM) después de una comida rica en nitratos. El nitrato es un factor ecológico que puede inducir cambios rápidos en la estructura y función de las comunidades polimicrobianas. Sin embargo, los efectos sobre la microbiota oral no se han estudiado en detalle, mientras que un número limitado de estudios previos a esta tesis indican que es probable que el nitrato sea beneficioso para la salud bucal. El objetivo de esta tesis es, por tanto, estudiar los cambios microbiológicos inducidos por nitratos e identificar posibles mecanismos de homeostasis generados por este compuesto, con el fin de determinar si el nitrato puede considerarse un prebiótico para la salud bucal. Un segundo objetivo fue aislar cepas reductoras de nitrato y probar su potencial probiótico in vitro. En el capítulo 1, se realizó un estudio in vitro para testar el efecto del nitrato 6,5 mM en comunidades orales cultivadas a partir de la saliva de 12 individuos sanos. En el capítulo 2, se obtuvieron 53 aislados de bacterias reductoras de nitrato y se probó el efecto de seis candidatos a probióticos en comunidades orales sanas cultivadas a partir de saliva de diferentes donantes con o sin nitrato 6,5 mM. En el capítulo 3, se estudió el efecto de un extracto de remolacha rico en nitrato sobre la acidificación oral después de un enjuague con azúcar en 24 individuos sin caries activas. Se tomaron sobrenadantes (capítulos 1 y 2) o muestras de saliva (capítulo 3) para mediciones de nitrato, nitrito, amonio, lactato y pH. Además, la composición bacteriana de la biopelícula in vitro y del pellet salivar se determinó usando secuenciación Illumina del rRNA 16S y/o qPCR del género nitratorreductor Rothia. Los datos demuestran que el nitrato estimula el crecimiento de los géneros beneficiosos Rothia y Neisseria en nuestro modelo in vitro, mientras que potencialmente disminuye las bacterias asociadas a la caries, la halitosis y la enfermedad periodontal. Además, los datos in vitro e in vivo presentados en esta tesis indican que el nitrato puede limitar o prevenir caídas de pH cuando los azúcares son fermentados por la microbiota oral, un mecanismo de resiliencia que podría ser estimulado por el consumo de extractos vegetales ricos en nitratos. Los principales mecanismos de amortiguación del pH por parte del nitrato son el uso de acido láctico durante la desnitrificación (observado tanto in vivo como in vitro) y durante la reducción de nitrito a amonio, así como la producción potencial de amoníaco (observado in vitro). En esta tesis, los efectos del nitrato se observaron después de períodos cortos, es decir, después de 5-9 h de incubación in vitro y 1-4 horas después de la ingesta del suplemento de nitrato in vivo. Los estudios futuros deberían centrarse en los efectos longitudinales de la ingesta diaria de nitratos. En el capítulo 2, se aislaron bacterias reductoras de nitrato pertenecientes a los géneros Rothia y Actinomyces. Una selección de aislados de Rothia aumentó el uso de lactato y la capacidad de reducción de nitratos de las comunidades bucales, lo que potencialmente beneficiaría la salud dental y la salud sistémica, respectivamente. Los datos in vitro e in vivo presentados en esta tesis sugieren que el nitrato puede modular la microbiota oral en aspectos que son beneficiosas para el huésped y, por lo tanto, podría considerarse una sustancia prebiótica para la microbiota oral. Además, los aislados reductores de nitratos pueden estimular los efectos beneficiosos del metabolismo del nitrato, sobre todo en personas con bajos niveles de estas bacterias. / [CA] S'ha estimat que obtenim més de les tres quartes parts del nitrat que ingerim de la fruita i la verdura. Els vegetals rics en nitrats inclouen verdures de fulla verda i uns certs tubercles (p. ex., remolatxes i raves). Les glàndules salivals concentren activament el nitrat plasmàtic, la qual cosa dona lloc a concentracions elevades de nitrat a la saliva (5 a 8 mm) després d'un menjar ric en nitrats. El nitrat és un factor ecològic que pot induir canvis ràpids en l'estructura i funció de les comunitats polimicrobianes. No obstant això, els efectes sobre la microbiota oral no s'han estudiat detalladament, mentre que un nombre limitat d'estudis previs a aquesta tesi indiquen que és probable que el nitrat siga beneficiós per a la salut bucal. L'objectiu d'aquesta tesi és, per tant, estudiar els canvis microbiològics induïts per nitrats i identificar possibles mecanismes d'homeòstasi generats per aquest compost, amb la finalitat de determinar si el nitrat pot considerar-se un prebiòtic per a la salut bucal. Un segon objectiu va ser aïllar soques reductores de nitrat i provar el seu potencial probiòtic in vitro. En el capítol 1, es va realitzar un estudi in vitro per a testar l'efecte del nitrat 6,5 mm en comunitats orals cultivades a partir de la saliva de 12 individus sans. En el capítol 2, es van obtindre 53 aïllats de bacteris reductors de nitrat i es va provar l'efecte de sis candidats a probiòtics en comunitats orals sanes cultivades a partir de saliva de diferents donants amb o sense nitrat 6,5 mm. En el capítol 3, es va estudiar l'efecte d'un extracte de remolatxa ric en nitrat sobre l'acidificació oral després d'un glopeig amb sucre en 24 individus sense càries actives. Es van prendre sobrenadants (capítols 1 i 2) o mostres de saliva (capítol 3) per a mesuraments de nitrat, nitrit, amoni, lactat i pH. A més, la composició bacteriana de la biopel·lícula in vitro i del pèl·let salivar es va determinar usant seqüenciació Illumina del RNAr 16S i/o qPCR del gènere nitratorreductor Rothia. Les dades demostren que el nitrat estimula el creixement dels gèneres beneficiosos Rothia i Neisseria en el nostre model in vitro, mentre que potencialment disminueix els bacteris associats a la càries, l'halitosi i la malaltia periodontal. A més a més, les dades in vitro i in vivo presentades en aquesta tesi indiquen que el nitrat pot limitar o previndre caigudes de pH quan els sucres són fermentats per la microbiota oral, un mecanisme de resiliència que podria ser estimulat pel consum d'extractes vegetals rics en nitrats. Els principals mecanismes d'amortiment del pH per part del nitrat són l'ús de àcid làctic durant la desnitrificació (observat tant in vivo com in vitro) i durant la reducció de nitrit a amoni, així com la producció potencial d'amoníac (observat in vitro). En aquesta tesi, els efectes del nitrat es van observar després de períodes curts, és a dir, després de 5-9 h d'incubació in vitro i 1-4 hores després de la ingesta del suplement de nitrat in vivo. Els estudis futurs haurien de centrar-se en els efectes longitudinals de la ingesta diària de nitrats. En aquesta tesi es van aïllar bacteris reductors de nitrat pertanyents als gèneres Rothia i Actinomyces. Una selecció d'aïllats de Rothia va augmentar l'ús de lactat i la capacitat de reducció de nitrats de les comunitats bucals, la qual cosa potencialment beneficiaria la salut dental i la salut sistèmica, respectivament. Les dades in vitro i in vivo presentats en aquesta tesi suggereixen que el nitrat pot modular la microbiota oral en aspectes que són beneficiosos per a l'hoste i, per tant, podria considerar-se una substància prebiòtica per a la microbiota oral. A més, els aïllats reductors de nitrats poden estimular els efectes beneficiosos del metabolisme del nitrat, sobretot en persones amb baixos nivells d'aquests bacteris. El nitrat i els bacteris reductors de nitrat són, per tant, components prometedors per a futurs productes de salut oral. / [EN] It has been estimated that we obtain over three quarters of dietary nitrate from vegetables and fruits. Nitrate-rich vegetable types include leafy greens and certain root vegetables (e.g., beetroots and radishes). The salivary glands actively concentrate plasma nitrate, leading to high salivary nitrate concentrations (5-8 mM) after a nitrate-rich meal. Nitrate is an ecological factor that can induce rapid changes in structure and function of polymicrobial communities. However, the effects on the oral microbiota have not been clarified, whilst a limited number of previous studies did indicate that nitrate is likely to be beneficial for oral health. The aim of this thesis was therefore to study nitrate-induced microbiome changes and identify potential mechanisms for nitrate-induced homeostasis, in order to determine if nitrate can be considered a prebiotic compound for oral health. A second aim was to isolate nitrate-reducing isolates and test their probiotic potential in vitro. In chapter 1, an in vitro study was set up testing the effect of 6.5 mM nitrate on oral communities grown from saliva of 12 healthy individuals. In chapter 2, fifty-three nitrate-reducing isolates were obtained and the effect of six probiotic candidates was tested on healthy oral communities grown from saliva of different donors with or without 6.5 mM nitrate. In chapter 3, the effects of nitrate-rich beetroot extracts on oral acidification after sugar rinsing was tested in 24 individuals without active caries. Supernatants (chapters 1 and 2) or saliva samples (chapter 3) were taken for nitrate, nitrite, ammonium, lactate and pH measurements. Additionally, the bacterial composition of in vitro biofilms and salivary pellets were determined using 16S rRNA gene Illumina sequencing and/or qPCR of the nitrate-reducing genus Rothia. We showed that nitrate stimulates the growth of the beneficial genera Rothia and Neisseria in our in vitro model, while potentially decreasing caries-, halitosis- and periodontal disease-associated bacteria. Additionally, the in vitro and in vivo data presented in this thesis indicate that nitrate can limit or prevent pH drops when sugars are fermented by the oral microbiota - a mechanism of resilience that could be stimulated by the consumption of nitrate-rich vegetable extracts. The main pH buffering mechanisms of nitrate were lactic acid usage during denitrification (observed both in vivo and in vitro) and during the reduction of nitrite to ammonium, as well as the potential production of ammonia (observed in vitro). In this thesis, the effects of nitrate were observed after short periods, i.e., after 5-9 h incubation in vitro and/or after 1-4 hours after nitrate supplement intake in vivo. Future studies should focus on the longitudinal effects of daily nitrate intake. In chapter 2, nitrate-reducing species belonging to the genera Rothia and Actinomyces were isolated. A selection of Rothia isolates increased lactate usage and nitrate reduction capacities of oral communities, potentially benefitting dental health and systemic health, respectively. The in vitro and in vivo data presented in the current thesis suggest that nitrate can modulate the oral microbiota in ways that are beneficial for the host and could thus be considered a prebiotic substance for the oral microbiota. Additionally, nitrate-reducing isolates can stimulate certain beneficial effects of nitrate metabolism. Nitrate and nitrate-reducing bacteria are thus promising components for future oral care products to prevent or treat oral diseases and this should be further investigated. / Rosier, BT. (2022). Nitrate as a Prebiotic and Nitrate-Reducing Bacteria as Probiotics for Oral Health [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/181578 / Compendio

Tesis por compendio / This thesis studies how to increase the expected satiating ability in dairy products analyzing the effect of the incorporation of certain ingredients. The study of food reformulation with high expected satiating ability contributes to moderate size meal choice and to regulate food intake. In this thesis, two factors influencing the satiating ability perception of semi-solid foods have been mainly studied: a) the increase in oral processing time that produces prolonged orosensory signals, and b) the increase in protein content. The increase in oral processing time is produced by, for example, higher consistencies, mouth-coating or creaminess sensations. These features can be modified by the addition of hydrocolloids. Native starch, modified starch, ¿-carrageenan and guar gum were added to increase the viscosity and consistency of milkshakes. Rheological characterization of the products revealed very different behavior patterns depending on the type of hydrocolloid. The addition of saliva in the studies showed modified sensory perceptions, so its inclusion is desirable and more realistic. The addition of hydroxypropyl methylcellulose (HPMC) and the effect of milk solids and cream in milk-based desserts was also studied. In addition to HPMC dominant effect on the temporality of perceptions, the presence of fat globules or milk solids modulated such perceptions. Yogurt was selected to increase protein content, adding twice the original amount of protein from different milk fractions, and starch. Yogurt with milk powder addition was the most accepted sample and it was closer to the 'ideal' characteristics of a satiating yogurt according to consumers, with a series of sensory attributes that contribute to the feeling of expected satiety. On the other hand, in vitro oral digestion showed that physically modified starch granules remain unchanged after the attack of ¿-amylase, and would be responsible for its consistency and creaminess. In addition, it was found that the sensation of astringency was undesirable; tribological studies allowed to interpret yogurt sensory characteristics. The lubricity of some samples did not reflect the difference in astringency between samples indicating that this was not a purely tactile perception caused by an increase in friction. The addition of physically modified starch significantly reduced friction coefficients values due to soluble starch polymers and starch granules preservation. All the strategies allowed to gain understanding on how to reformulate food increasing its satiating ability perception. Such strategies could be extrapolated to other food categories, what broadens the scope of the results obtained in this thesis. / La presente tesis estudia cómo aumentar las expectativas de la capacidad saciante en productos lácteos analizando el efecto de la incorporación de determinados ingredientes. El estudio de la reformulación de alimentos con alta expectativa sobre su capacidad saciante contribuye a moderar la elección de las raciones de alimentos y a regular la ingesta. En la presente tesis se han estudiado principalmente dos de los factores que más influyen en la percepción de capacidad saciante de alimentos semisólidos: a) el aumento en el tiempo de procesado oral que produce saciedad debido a una prolongación de las señales orosensoriales, y b) el alto contenido en proteínas. El aumento del tiempo de procesado oral lo producen, por ejemplo, mayores consistencias, recubrimiento oral o sensaciones de cremosidad, que son características que se pueden modificar por adición de hidrocoloides. Se emplearon almidón nativo y modificado, goma guar y ¿-carragenato para aumentar la viscosidad y consistencia de batidos lácteos. La caracterización reológica de los productos reveló patrones de comportamiento muy diferentes dependiendo del tipo de hidrocoloide. La adición de la saliva en los estudios demostró modificar las percepciones sensoriales, por lo que su inclusión resulta recomendable y más realista. Se estudió también la adición de hidroxipropilmetilcelulosa (HPMC) y el efecto de dos componentes normales en productos lácteos (sólidos lácteos y nata). Además del efecto dominante del HPMC en la temporalidad de las percepciones, la presencia de glóbulos de grasa o sólidos de la leche modularon dichas percepciones. Para aumentar el contenido en proteína se seleccionó como alimento el yogur, al que se añadió el doble de la cantidad de proteína de distintas fracciones lácteas y almidón. El yogur con el doble de leche en polvo fue globalmente el más aceptado y más cercano a las características "ideales" según los consumidores, con una serie de atributos sensoriales que contribuyen a la sensación de saciedad esperada. Por otro lado, la digestión oral in vitro mostró que los gránulos de almidón procedentes de almidón físicamente modificado permanecen inalterados después del ataque de la ¿-amilasa de la saliva, y serían los responsables de la mayor consistencia y cremosidad. Adicionalmente, se encontró que la sensación de astringencia resultaba negativa; los estudios tribológicos permitieron interpretar las características sensoriales. Las propiedades lubricantes de algunas muestras no reflejaron la diferencia de astringencia sensorial indicando que ésta no era una percepción puramente táctil causada por un aumento en la fricción. La adición de almidón modificado físicamente redujo significativamente los valores de los coeficientes de fricción relacionados con los polímeros de almidón solubles y la conservación de gránulos en el almidón. Todas las estrategias abordadas permitieron comprender y ahondar en los conocimientos que conducen a cómo reformular un alimento para aumentar la percepción de saciedad. Dichas estrategias son extrapolables a otras categorías de alimentos lo que amplía el alcance de los resultados obtenidos en la presente tesis. / La present tesi estudia com augmentar les expectatives de la capacitat saciant en productes lactis analitzant l'efecte de la incorporació de determinats ingredients. L'estudi de la reformulació d'aliments amb alta expectativa sobre la seua capacitat saciant contribueix a moderar l'elecció de les racions d'aliments i a regular la ingesta. En la present tesi s'han estudiat principalment dos dels factors que més influeixen en la percepció de capacitat saciant d'aliments semisòlids: a) l'augment en el temps de processat oral que produeix sacietat a causa d'una prolongació dels senyals orosensorials, i b) l'alt contingut en proteïnes. L'augment del temps de processat oral el produeixen, per exemple, majors consistències, recobriment oral o sensacions de cremositat, que són característiques que es poden modificar per addició d'hidrocol·loïdes. S'emprà midó natiu i modificat, goma guar i ¿-carragenat per augmentar la viscositat i consistència de batuts lactis. La caracterització reològica dels productes revelà patrons de comportament molt diferents depenent del tipus d'hidrocol·loïde. L'addició de la saliva en els estudis demostrà modificar les percepcions sensorials, per la qual cosa la seua inclusió resulta recomanable i més realista. S'estudià també l'addició d'hidroxipropilmetilcel·lulosa (HPMC) i l'efecte de dos components normals en productes lactis (sòlids lactis i nata). A més de l'efecte dominant del HPMC en la temporalitat de les percepcions, la presència de glòbuls de greix o sòlids de la llet van modular les citades percepcions. Per augmentar el contingut en proteïna es va seleccionar com a aliment el iogurt, a què s'afegí el doble de la quantitat de proteïna de diverses fraccions làcties i midó. El iogurt amb el doble de llet en pols fou globalment el més acceptat i més pròxim a les característiques "ideals" segons els consumidors, amb una sèrie d'atributs sensorials que contribueixen a la sensació de sacietat esperada. D'altra banda, la digestió oral in vitro mostrà que els grànuls de midó procedents de midó físicament modificat romanen inalterats després de l'atac de l'¿-amilasa de la saliva, i serien els responsables de la major consistència i cremositat. Addicionalment, es trobà que la sensació d'astringència resultava negativa; els estudis tribològics permeteren interpretar les característiques sensorials. Les propietats lubricants d'algunes mostres no reflectiren la diferència d'astringència sensorial indicant que esta no era una percepció purament tàctil causada per un augment en la fricció. L'addició de midó modificat físicament reduí significativament els valors dels coeficients de fricció relacionats amb els polímers de midó solubles i la conservació de grànuls en el midó. Totes les estratègies abordades permeteren comprendre i aprofundir en els coneixements que condueixen a com reformular un aliment per augmentar la percepció de sacietat. Les citades estratègies són extrapolables a altres categories d'aliments, cosa que amplia l'abast dels resultats obtinguts en la present tesi. / Morell Esteve, P. (2017). Diseño de alimentos lácteos con capacidad saciante. Relación entre estructura, procesamiento oral y percepción [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/89093 / Compendio

CONTEXTE : Le cancer colorectal (CCR) reste l’un des cancers les plus diagnostiqués au Canada et dans le monde. Malgré les efforts en matière de dépistage, la moitié des cancers colorectaux sont encore diagnostiqués à des stades avancés de la maladie, réduisant ainsi les chances de survie à ce cancer. La prévention du CCR reste encore primordiale et les efforts doivent être maintenus pour l’identification de ses facteurs de risque modifiables. Durant cette dernière décennie, on a assisté à un cumul d’évidence sur l’association entre l’enrichissement colorectal en une bactérie spécifique, Fusobacterium nucleatum (F. nucleatum), et le CCR. Ce qui a ressuscité le débat sur l’association même entre la maladie parodontale (MP) et le CCR, puisque F. nucleatum est non seulement une bactérie commensale de la bouche, mais surtout l’une des principaux agents pathogènes de la MP. Des études épidémiologiques antérieures avaient en effet déjà suggéré que le la MP serait associée au risque de CCR, mais leurs résultats sont restés non concluants à cause de leurs défaillances méthodologiques. La diffusion des bactéries parodontopathogènes, comme F. nucleatum, et de leurs produits de virulence de la bouche vers le côlon et la libération des médiateurs d’inflammation chronique dans la circulation générale sont les mécanismes incriminés dans l’association entre la MP et le CCR. OBJECTIFS: 1) Vérifier l’association entre la MP et le CCR; 2) Synthétiser la littérature sur l’association entre la détection de la bactérie F. nucleatum dans le côlon et le CCR et; 3) Vérifier la faisabilité d’investiguer l’expression concomitante de F. nucleatum, dans les deux milieux buccal et colorectal, en présence de lésions néoplasiques colorectales. Ces deux derniers objectifs sont un préambule à l’investigation du rôle des bactéries causales de la MP, précisément F. nucleatum, comme mécanisme d’association entre la MP et le CCR. MÉTHODES: Nous avons entrepris trois projets de recherche distincts qui répondent aux trois objectifs respectifs de recherche cités ci-dessus: 1) Une étude cas-témoins à base populationnelle (Projet COLDENT) a inclut 348 cas incidents de CCR et 310 témoins qui lui sont appariés dans des catégories d’âge et de sexe. Des données ont été collectées sur l’exposition à la MP et aux principaux facteurs de risque du CCR; 2) Une revue systématique avec méta-analyse d’études observationnelles ayant investigué la bactérie, F. nucleatum, dans des échantillons colorectaux, chez des cas de CCR et des témoins exempts de lésions cancéreuses ou précancéreuses colorectales; 6 et 3) Une étude pilote cas-témoins à base hospitalière, auprès de 22 cas de néoplasies colorectales et 21 témoins, examinés par coloscopie. Des échantillons de salive et des biopsies de muqueuse colorectale, ainsi que des données sur l’exposition à la MP et aux principaux facteurs de risque du CCR ont été collectées. RÉSULTATS: Les résultats de notre étude cas-témoins populationnelle suggèrent que le taux d’incidence de CCR était augmenté chez les personnes avec un historique positif de MP comparé aux personnes sans historique de MP, en ajustant pour la majorité des facteurs de confusion potentiels (Rapport des taux ajusté = 1,45 ; Intervalle de confiance (IC) à 95 % : 1,04-2,01). Les résultats de notre revue systématique incluant 24 études observationnelles ont montré que dans la majorité des études où la charge de F. nucleatum dans les échantillons colorectaux était quantifiée, celle-ci était significativement plus élevée chez les cas de CCR que chez les témoins. Les résultats de la méta-analyse de 12 études observationnelles qui ont rapporté le pourcentage des échantillons colorectaux où F. nucleatum était détecté montrent une association significativement positive entre la détection de la bactérie F. nucleatum dans le côlon et le CCR (Rapport de cotes combiné = 8,3; IC à 95 % = 5,2-13; Indice d’hétérogénéité I2 = 26,3 %; p pour hétérogénéité = 0,18). Enfin, les résultats préliminaires de l’étude de faisabilité appuient la faisabilité et l’utilité de mener une étude subséquente pour investiguer F. nucleatum dans la salive et le côlon en présence de néoplasies colorectales afin de vérifier son rôle dans l’association entre la MP et le CCR, mais des ajustements doivent être apportés à la méthodologie initiale. CONCLUSION: Nos résultats suggèrent que l’exposition à la MP peut augmenter le risque de CCR, bien qu’on ne puisse encore confirmer de lien causal. On espère que ce travail sensibilise les cliniciens, décideurs en santé et public à l’importance de prévenir et de contrôler les maladies buccodentaires, notamment la MP, pour la prévention même de maladies systémiques encore plus fatales, tels que le CCR. / BACKGROUND: Colorectal cancer (CRC) remains one of the most diagnosed cancers in Canada and around the world. Despite screening efforts, half of colorectal cancers are still diagnosed at advanced stages of the disease, thus reducing the chances of survival from this cancer. Prevention of CRC still remains essential, and efforts must be maintained to identify its modifiable risk factors. Over the past decade, there has been an accumulation of evidence on the association between colorectal enrichment in a specific bacterium, Fusobacterium nucleatum (F. nucleatum), and CRC. This has sparked debate on the association between periodontal disease (PD) and CRC, since F. nucleatum is not only a commensal bacterium of the mouth, but above all one of the main pathogens of PD. Previous epidemiological studies had indeed already suggested that PD would be associated with the risk of CRC, but their results remained inconclusive because of their methodological flaws. Dissemination of periodontal pathogen bacteria such as F. nucleatum, and of their virulence products, from the mouth to the colon and the release of chronic inflammatory mediators into the bloodstream are suggested as the mechanisms involved in the association between PD and CRC. OBJECTIVES: 1) To verify the association between PD and CRC; 2) To synthesize the literature on the association between the detection of the bacterium F. nucleatum in the colon and CRC; and 3) To verify the feasibility of investigating the concomitant expression of F. nucleatum, in both oral and colorectal environments, in the presence of colorectal neoplasms. These last two objectives are a preamble to the investigation of the role of the causative bacteria of PD, specifically F. nucleatum, as a mechanism of association between PD and CRC. METHODS: We undertook three separate research projects that address the three respective research objectives listed above: 1) A population-based case-control study (COLDENT Project) included 348 incident cases of CRC and 310 age and sex frequency-matched controls. Data were collected on PD and on major risk factors for CRC; 2) A systematic review with meta-analysis of observational studies that have investigated the bacterium F. nucleatum in colorectal samples, in CRC cases and colorectal cancerous or precancerous-free controls; and 3) A pilot hospital-based case-control study, with 22 cases of colorectal neoplasms and 21 controls, examined by colonoscopy. Saliva samples and colorectal mucosal biopsies as well as data on exposure to PD and major risk factors for CRC were collected. 8 RESULTS: The results of our population-based case-control study suggest that the rate of CRC was increased in people with a positive history of PD compared to people without a history of PD, adjusting for the majority of potential confounders (adjusted rate ratio = 1.45; 95% Confidence interval (CI): 1.04-2.01). The results of our systematic review including 24 observational studies showed that in most studies where the load of F. nucleatum in colorectal samples was quantified, it was significantly higher in CRC cases than in controls. The results of the meta-analysis of 12 observational studies that reported the percentage of colorectal samples where F. nucleatum was detected show a significantly positive association between the detection of the bacterium F. nucleatum in the colon and CRC (pooled odds ratio = 8.3; 95% CI = 5.2-13; heterogeneity index I 2 = 26.3%; p for heterogeneity = 0.18). Finally, the preliminary results of the feasibility study support the feasibility and utility of conducting a subsequent study to investigate F. nucleatum in saliva and colon in the presence of colorectal neoplasms to verify its role in the association between PD and CCR, but adjustments need to be made to the original methodology. CONCLUSION: Our results suggest that exposure to PD may increase the risk of CRC, although a causal relationship cannot yet be confirmed. It is hoped that this work will sensitize clinicians, health policy makers and the public to the importance of preventing and controlling oral diseases, including PD, for the prevention of even more fatal systemic diseases, such as CRC.

This thesis explores the potential of Differential Dynamic Microscopy (DDM) as a rapid diagnostic platform with a simplified sample preparation protocol, suitable for both laboratory use and future point-of-use (POU) applications. Focused initially on COVID-19 diagnostics, our study addresses the need for novel techniques capable of quantifying both antibody and viral loads, crucial for comprehensive patient profiling. Traditional assays like PCR and ELISA, while sensitive, are hindered by complexity and expensive equipment requirements. Two distinct DDM-based diagnostic approaches are detailed: protein biomarker quantification and Virus-Like Particle (VLP) quantification. The protein biomarker assay utilizes fluorescence imaging of probe beads, demonstrating quantification capabilities for antibodies in saliva and serum with minimal manipulation and short incubation times. This assay is adaptable to different antibodies and diseases, utilizing widely available laboratory equipment. Conversely, the VLP assay employs dark-field imaging of gold nanoparticle sensors to detect VLPs, achieving a low limit of detection in buffer and salivary samples without amplification or labeling steps. These methodologies highlight DDM's potential for broad laboratory adoption and future POC applications through equipment miniaturization. We present a versatile diagnostic platform poised to revolutionize medical diagnostics, simplifying procedures and accelerating disease detection to improve patient outcomes across diverse healthcare settings. High-throughput implementation in laboratories, including remote and resource-limited regions, is anticipated. Future directions involve clinical validation studies for assay sensitivity and specificity, exploring multiplexing capabilities, and assessing vaccine efficacy using DDM-based assays. The ongoing pursuit of miniaturized equipment aims to further enable POU applications, enhancing accessibility and efficacy in disease diagnostics and monitoring. / Cette thèse explore le potentiel de la Microscopie Dynamique Différentielle (MDD) en tant que plateforme de diagnostic rapide avec un protocole simplifié de préparation des échantillons, adaptée à une utilisation en laboratoire ainsi qu'à des applications futures sur site. Axée initialement sur le diagnostic de la COVID-19, notre étude répond au besoin de techniques novatrices capables de quantifier à la fois les anticorps et les charges virales, essentielles pour un profilage complet des patients. Les méthodes traditionnelles telles que la PCR et l'ELISA, bien que sensibles, sont entravées par leur complexité et les exigences onéreuses en équipement. Deux approches diagnostiques distinctes basées sur la MDD sont détaillées : la quantification des biomarqueurs protéiques et la quantification des particules pseudo-virales (PPV). L'essai pour la détection des biomarqueurs protéiques utilise l'imagerie par fluorescence des billes sondes, démontrant des capacités de quantification des anticorps dans la salive et le sérum avec une manipulation minimale et des temps d'incubation courts. Cette méthode est adaptable à différents anticorps et maladies, utilisant des équipements de laboratoire largement disponibles. En revanche, l'essai de détection des PPV utilise l'imagerie en champ sombre des capteurs à base de nanoparticules d'or pour détecter les PPV, atteignant une faible limite de détection dans les échantillons tampon et salivaires sans étapes d'amplification ni d'étiquetage. Ces méthodologies mettent en lumière le potentiel de la MDD pour une adoption étendue en laboratoire et des futures applications sur site grâce à la miniaturisation de l'équipement. Nous présentons une plateforme de diagnostic polyvalente prête à révolutionner le diagnostic médical, simplifiant les procédures et accélérant la détection des maladies pour améliorer les résultats des patients dans divers environnements de soins de santé. Une mise en œuvre à haut débit dans les laboratoires, y compris dans les régions éloignées et à ressources limitées, est anticipée. Les orientations futures incluent des études cliniques de validation pour la sensibilité et la spécificité des essais de détection, l'exploration des capacités de multiplexage, et l'évaluation de l'efficacité des vaccins à l'aide de la technique MDD. La recherche continue d'équipements miniaturisés vise à faciliter davantage les applications sur site, améliorant ainsi l'accessibilité et l'efficacité dans le diagnostic et le suivi des maladies.

Background. The current understanding on caries and enamel developmental defects prevalence and frequency, caries risk, health-related quality life (HRQoL) and stress response in young children with cleft lip and/or palate (CL/P) are sparse. In this thesis these aspects were investigated in 5- and 10- year-old children with CL/P in comparison to non-cleft children in the same ages. Design. The studies in this thesis have a cross-sectional case-control design. Participants. The study group consisted of 139 children with CL/P (80 children aged 5 years and 59 aged 10 years) and 313 non-cleft controls (144 children aged 5 years and 169 aged 10 years). Method. Caries was scored according to International Caries Detection and Assessment System (ICDAS II) and developmental enamel defects were measured as presence and frequency of hypoplasia and hypomineralization. Oral hygiene was assessed using Quigley-Hein plaque index. Stimulated saliva samples were analyzed for mutans streptococci, lactobacilli, buffering capacity and secretion rates. Information regarding children’s oral hygiene routines, dietary habits and fluoride exposure were collected with questionnaires. Caries risk was evaluated with algorithm-based software, Cariogram while HRQoL was perceived with KIDSCREEN-52. Stress response was analyzed with cortisol concentration in saliva at three different time points using a commercial competitive radioimmunoassay. Results. Caries prevalence (36% versus 18%) and caries frequency (1.2 dmfs versus 0.9 dmfs) was significantly higher in 5-year-old children with CL/P in comparison to non-cleft controls. In 10-yearolds no significant difference was found between children with CL/P and non-cleft controls in caries prevalence (47% versus 38%) or in caries frequency (0.7 DMFS versus 0.5 DMFS). Children with CL/P had significantly higher prevalence of enamel defects, higher counts of salivary lactobacilli and less good oral hygiene. The odds of being categorized with high caries risk were elevated in children with CL/P. Children with CL/P had similar HRQoL and salivary cortisol concentrations as non-cleft controls. However, 10-year-old boys with CL/P had significantly higher cortisol concentrations in the evening than non-cleft boys. Conclusions. Preschool children with CL/P seem to have more caries in the primary dentition than non-cleft controls. Children with CL/P had increased odds of being categorized as high caries risk individuals compared to controls. Some of the contributing factors seem to be higher prevalence of enamel defects, impaired oral hygiene and elevated salivary lactobacilli. Furthermore, as measured with the help of cortisol concentrations in saliva, children with CL/P were not more stressed than noncleft controls and their HRQoL was comparable to a European norm population. It appears that regular comprehensive preventive oral care in children with CL/P is effective in preventing caries development in permanent teeth. However, children with CL/P are at risk of caries development and preventive oral care should be implemented and started earlier than today.

<p>Uvod: Pojam &quot;sekretor&rdquo; ili &bdquo;nesekretor&quot; se odnosi na sposobnost pojedinca da luči antigene krvnih grupa ABO sistema u telesnim tečnostima. Određivanje ABO krvne grupe i sekretornog statusa, testom inhibicije hemaglutinacije pljuvačke i Lewis fenotipa na eritrocitima su važni u kliničkoj i forenzičkoj medicini, u odnosu na etiopatogenezu mnogih bolesti. Nesekretorstvo ABO krvnogrupne supstance je udruženo sa če&scaron;ćom pojavom autoimunih inflamatornih oboljenja među kojima su i seronegativne spondiloartropatije. Veća učestalost seronegativnih spondiloartropatija među osobama koje imaju HLA-B27 antigen predstavlja polaznu osnovu stanovi&scaron;ta da genetski faktori u kombinaciji sa faktorima sredine utiču na pojavu seronegativnih spondiloartropatija u genetski predisponiranih individua. Teza istražuje značaj testa inhibicije hemaglutinacije plijuvačke i određivanja ABO i Lewis fenotipa na eritrocitim u dijagnostici seronegativnih spondiloartropatija. Ciljevi i hipoteze: Cilj istraživanja je bio da se utvrdi učestalost nesekretora i sekretora ABO krvnogrupne supstance i Lewis fenotipa u grupi obolelih od seronegativnih spondiloartropatija i izvr&scaron;iti poređenje rezultata u odnosu na grupu zdravih ispitanka. Pretpostavljeno je da postoji značajno veći broj nesekretora ABO krvnogrupne supstance u obolelih od seronegativnih spondiloartropatija u odnosu na zdrave ispitanike. Pretpostavljeno je i da postoji značajno veća učestalost nesekretora ABO krvnogrupne supstance u obolelih od seronegativnih spondiloartropatija sa negativnim HLA-B27 antigenom. Pretpostavljeno je da kod osoba obolelih od seronegativnih spondiloartropatija dolazi do promene Lewis fenotipa na eritrocitima u odnosu na sekretorni status u pljuvačci. Metode: Sprovedena je longitudinalna prospektibvna studija. Ispitanici stariji od &scaron;est godina oba pola podeljeni su u dve randomizovane grupe. Eksperimentalnu grupu sačinjavalo je 110 ispitanika sa dijagnozom oboljenja iz grupe seronegativnih spondiloartropatija. Kontrolnu grupu sačinjavalo je 103 dobrovoljna davaoca krvi, bez dijaghnoze oboljenja iz grupe seronegativnih spondiloartropatija. Ispitanicima kontrolne i eksperimentalne grupe određena je pripadnosti ABO krvnogrupnom sistemu, sekretorni status i Lewis fenotip, dok je osobama eksperimentalne grupe određen i fenotip HLA-B27. Uključujući kriterijumi osim navedenog bili su da ispitivane osobe ženskog pola nisu trudinice i da ispitanici obe grupe nisu primali transfuziju krvi tri meseca pre uključivanja u istraživanje. Rezultati: Rezultati istraživanja ukazuju da nesekretori ABO krvnogrupne supstance imaju 1,63 puta veći rizik (veću učestalost), oboljevanja od seronegativnih spondiloartropatija u odnosu na zdravu populaciju ispitanika, kao i da smanjena ekspresija Lewis (b) antigena predstavlja doprinoseći faktor razvoja seronegativnih spondiloartropatija. Ustanovljeno je da pod uticajem seronegativnih spondiloartropatija dolazi do izmene Lewis fenotipa na eritrocitima obolelih. Verovatnoća dokazivanja oboljenja iz grupe seronegativnih spondilartropatija među obolelima veća je za 11% kod nesekretora fenotipa HLA-B27- u odnosu na obolele nesekretore fenotipa HLA-B27+. Zaključak: Sekretorni status i Lewis fenotip predstavljaju zasebne dijagnostičke biohemijske markere nezavisne od HLA-B27 antigena koji doprinose ranijem otkrivanju osoba koje imaju predispoziciju razvoja oboljenja iz grupe seronegativnih spondiloartropatija.</p> / <p>Introduction:The term secretory state referes to ability of individual to secrete ABO blood group antigens in body fluids. Determination of the ABO blood group antigens and secretory status by hemagglutionation inhibition test using saliva as well as Lewis phenotype on erythrocytes are important in clinical and forensic medicine, in relation to the etiopathogenesis of many diseases. Non secretory status of ABO blood groupantegens is related with higher incidence of autoimmune inflammatory disease which include seronegative spondyloarthropathyes. Increased frequency of seronegative spondyloarthropathies among people who have the HLA-B27 antigen is starting point of the view that genetic factors in combination with environmental factors influence the occurence of seronegative spondyloarthropathies in genetically predisposed individuals. The tesis explores the significance of the hemagglutionation inhibition test of saliva and determination of ABO and Lewis antigens in diagnostics of seronegativespondyloarthropathyes. Goals and hypothesis: The aim of this study was to determine the frequency of non secretors and secretors of ABO blood group antigens and Lewis phenotype in a group of patient with seronegative spondyloarthropathies and make comparsion to the healthy examined group. It was assumed that there is a significantly higher number of non secretors of ABO blood group antigens among the patients with seronegative spondyloarthropathies compared to healthy examined persons. It was assumed that there is a significantly higher number of non secretors of ABO blood group antigens among the patients with seronegative spondyloarthropathies who do not have HLA-B27 antigen. It was assumed that in patients with seronegative spondyloarthropathies Lewis phenotype on erythrocits can be changed in relation to the secretory status in the saliva. Methods: We performed a prospective longitudinal study. Respondants older than six years of both gender were divided into two randomized groups. Experimental group is consisted of 110 patients diagnosed with seronegative spondyloarthropathy. The control group consisted of 103 blood donors who did not have diagnosed disease from the group of seronegative spondyloarthropathies. To the subjects of the control and experimental groups was determined ABO blood group antigens, secretory status and Lewis phenotype, while to the subjects of experimental group also was designated HLA-B27 phenotype. Including criteria other than the above were that among female respondants were not pregnant, and that both groups of respondants did not receive a blood transfusion three months before joining the study. Results: The resuts of the study showed that non secretors of ABO blood group antigens have a 1,63 times higher rise (higher incidence) of developing disease from the group of seronegative spondyloarthropathies compared to a healthy population of subjects, as well as that decreased expression of Lewis antigens (b) represents a contributing factor for development of seronegativespondyloarthropathies. It was found that under the influence of seronegative spondyloarthropathies there are changes in Lewis phenotype on erythrocytes of patients. It was found that under the influence of seronegative spondyloarthropathies there are changes in Lewis phenotype on erythrocytes of patients. Probability for confirmation seronegative spondyloarthropathies is higher for 11% among non secretors who have HLA-B27 negative phenotype in comparison to non secretors who have HLA-B27 positive phenotype. Conclusion: Secretory status and Lewis phenotype are separate diagnostic biochemical markers independent of HLA-B27 antigen that contribute to the early detection of people who have a predisposition of the disease from the group of seronegative spondyloarthropathies.</p>

El texto completo de este trabajo no está disponible en el Repositorio Académico UPC por restricciones de la casa editorial donde ha sido publicado. / PURPOSE: To assess the long-term effect of a 2% aqueous chlorhexidine (CHX) solution on bond strength to artificially eroded dentin compared to sound dentin. MATERIALS AND METHODS: Flat mid-coronal dentin surfaces of extracted third molars (n = 28) were subjected only to grinding with a 600-grit SiC paper for 1 min (sound dentin S, n = 14) or additionally to erosive pH cycling with a cola-based soft-drink (eroded dentin E, n = 14). After acid etching, rinsing, and air drying, S and E were rehydrated with 1.5 μl of 2% CHX (S2%, n = 7; E2%, n = 7) or of distilled water (control SC, n = 7; EC, n = 7). Composite buildups were incrementally constructed with Filtek Z350 following Adper Single Bond 2 application. Specimens were sectioned into beams, which were subjected to microtensile testing immediately or after 6 or 12 months of aging. Fractured surfaces were observed under a digital microscope (50X magnification). Microtensile bond strength (μTBS) (MPa) was analyzed by three-way ANOVA and Tukey's tests (α = 0.05) and failure mode by the Kruskal-Wallis test (α = 0.05). RESULTS: Compared to sound dentin, eroded dentin was consistently related to lower μTBS. Immediately and after 12-month aging, the effect of CHX was insignificant, but it was significant after 6-month aging, when it conserved the bond strength to both eroded and sound dentin. The percentage of adhesive and mixed failures were equivalent, and significantly more frequent than cohesive failures, whether in dentin or in composite. CONCLUSION: The 2% CHX effect on bond strength conservation to both eroded and sound dentin was not found to be persistent. / Revisión por pares

<p>Увод: Хронична бубрежна инсуфицијенција (ХБИ) се дефинише као структурно или функционално оштећење бубрега у трајању од најмање три месеца и/или смањење јачине гломеруларне филтрације (ЈГФ) испод 60 мл/мин/1.73м2. У доступној литератури постоје различити подаци о присуству оралних манифестација код пацијената са ХБИ у квантитативном и квалитативном погледу. Стање бубрежне дисфункције праћено је променама у протоку и саставу пљувачке што је у последњој деценији допринело испитивању клиничких и лабораторијских показатеља бубрежне болести. Циљ: Циљ студије је био да се испита објективно стање оралне слузокоже, вредности рН, сијалометрије, концентрације урее, креатинина и секреторног имуноглобулина А пљувачке као и орални микробиолошки статус код пацијената са ХБИ. Материјал и методе: Узорак је био сачињен од 50 предијализних (31 мушкарца и 19 жена просечне старости 59,06&plusmn;14,30) и 25 хемодијализних пацијената (18 мушкараца и 7 жена просечне старости 54,92&plusmn;13,60) са постављеном дијагнозом ХБИ, заједно са 25 системски здравих испитаника компарибилних по полу и старости. Поред клиничког прегледа усне дупље спроведен је тест витроадхезије, одређивање интензитета саливације, рН вредности пљувачке и индекса крварења из интерденталне папиле (PBI). На узорцима сакупљене пљувачке, уз помоћ аутоматизованог система Beckman Coulter АУ480 спроведено је лабораторијско одређивање урее и креатинина методом спектрофотометрије и секреторног имуноглобулина А методом имунотурбидиметрије. За микробилошко испитивање коришћен је брис језика и техника оралног испирка. Резултати: Нису утврђене статистички значајне разлике између група према демографско-социјалним подацима. Предијализни испитаници су имали значајно веће присуство промена оралне слузокоже и оралних симптома. Просечне вредности клиренса креатинина су биле значајно мање код оболелих испитаника са бледилом оралне слузокоже, уремичним задахом, ксеростомијом и измењеним осећајем укуса у поређењу са испитаницима без наведених промена. Код предијализних су утврђене значајно смањене вредности сијалометрије према контролним групама и повећане pH вредности према групи здравих испитаника. Просечне концентрације урее и креатинина су се статистички значајно разликовале између испитиваних група. Умерена позитивна корелација је утврђена између серумских и пљувачних концентрација урее и креатинина код предијализних и креатинина код хемодијализних. Према просечним вредностима секреторног имуноглобулина А није било разлика између група. Код пацијената са ХБИ утврђено је значајно веће присуство гљива из рода Candida са предоминацијом non-albicans Candida врста. Закључак: Резултати истраживања указују на важност утврђивања клиничких карактеристика усне дупље код предијализних пацијената. Интензитет саливације, pH вредност и пљувачне концентрације уремијских токсина могу бити поуздани маркери бубрежног оштећења. Једноставан и неинвазиван приступ приликом узорковања пљувачке и поузданост лабораторијске анализе треба да допринесу широј примени пљувачке као компетитивним дијагностичким флуидом серуму. Техника оралног испирка је прецизна квантитативна метода за одређивање степена гљивичне колонизације.</p> / <p>Uvod: Hronična bubrežna insuficijencija (HBI) se definiše kao strukturno ili funkcionalno oštećenje bubrega u trajanju od najmanje tri meseca i/ili smanjenje jačine glomerularne filtracije (JGF) ispod 60 ml/min/1.73m2. U dostupnoj literaturi postoje različiti podaci o prisustvu oralnih manifestacija kod pacijenata sa HBI u kvantitativnom i kvalitativnom pogledu. Stanje bubrežne disfunkcije praćeno je promenama u protoku i sastavu pljuvačke što je u poslednjoj deceniji doprinelo ispitivanju kliničkih i laboratorijskih pokazatelja bubrežne bolesti. Cilj: Cilj studije je bio da se ispita objektivno stanje oralne sluzokože, vrednosti rN, sijalometrije, koncentracije uree, kreatinina i sekretornog imunoglobulina A pljuvačke kao i oralni mikrobiološki status kod pacijenata sa HBI. Materijal i metode: Uzorak je bio sačinjen od 50 predijaliznih (31 muškarca i 19 žena prosečne starosti 59,06&plusmn;14,30) i 25 hemodijaliznih pacijenata (18 muškaraca i 7 žena prosečne starosti 54,92&plusmn;13,60) sa postavljenom dijagnozom HBI, zajedno sa 25 sistemski zdravih ispitanika komparibilnih po polu i starosti. Pored kliničkog pregleda usne duplje sproveden je test vitroadhezije, određivanje intenziteta salivacije, rN vrednosti pljuvačke i indeksa krvarenja iz interdentalne papile (PBI). Na uzorcima sakupljene pljuvačke, uz pomoć automatizovanog sistema Beckman Coulter AU480 sprovedeno je laboratorijsko određivanje uree i kreatinina metodom spektrofotometrije i sekretornog imunoglobulina A metodom imunoturbidimetrije. Za mikrobiloško ispitivanje korišćen je bris jezika i tehnika oralnog ispirka. Rezultati: Nisu utvrđene statistički značajne razlike između grupa prema demografsko-socijalnim podacima. Predijalizni ispitanici su imali značajno veće prisustvo promena oralne sluzokože i oralnih simptoma. Prosečne vrednosti klirensa kreatinina su bile značajno manje kod obolelih ispitanika sa bledilom oralne sluzokože, uremičnim zadahom, kserostomijom i izmenjenim osećajem ukusa u poređenju sa ispitanicima bez navedenih promena. Kod predijaliznih su utvrđene značajno smanjene vrednosti sijalometrije prema kontrolnim grupama i povećane pH vrednosti prema grupi zdravih ispitanika. Prosečne koncentracije uree i kreatinina su se statistički značajno razlikovale između ispitivanih grupa. Umerena pozitivna korelacija je utvrđena između serumskih i pljuvačnih koncentracija uree i kreatinina kod predijaliznih i kreatinina kod hemodijaliznih. Prema prosečnim vrednostima sekretornog imunoglobulina A nije bilo razlika između grupa. Kod pacijenata sa HBI utvrđeno je značajno veće prisustvo gljiva iz roda Candida sa predominacijom non-albicans Candida vrsta. Zaključak: Rezultati istraživanja ukazuju na važnost utvrđivanja kliničkih karakteristika usne duplje kod predijaliznih pacijenata. Intenzitet salivacije, pH vrednost i pljuvačne koncentracije uremijskih toksina mogu biti pouzdani markeri bubrežnog oštećenja. Jednostavan i neinvazivan pristup prilikom uzorkovanja pljuvačke i pouzdanost laboratorijske analize treba da doprinesu široj primeni pljuvačke kao kompetitivnim dijagnostičkim fluidom serumu. Tehnika oralnog ispirka je precizna kvantitativna metoda za određivanje stepena gljivične kolonizacije.</p> / <p>Introduction: Chronic kidney disease (CKD) is defined as structural and functional kidney damage for a period of at least three months and/or reduction of glomerular filtration rate (GFR) under 60 ml/min/1.73m2. There are different data in the available literature in term of quantitative and qualitative presence of the oral manifestation in patients with CKD. Kidney dysfunction is accompanied by changes in the salivary flow and composition, which is in the last decade contributed by examination of clinical and laboratory markers of renal disease. Aim: The aim of the study was to examine condition of oral mucosa, pH value, salivary flow rate, concentration of salivary urea, creatinine, secretory immunoglobulin A and oral microbiological status in patients with CKD. Materials and Methods: The sample was consisted of 50 predialysis (31 males and 19 females, mean age 59,06&plusmn;14,30) and 25 hemodialysis patients (18 males and 7 females, mean age 54,92&plusmn;13,60) with a diagnosis of CKD, along with 25 age and gender matched healthy controls. In addition of clinical examination, tongue blade adhesion test, sialometry, salivary pH test and determination of papilla bleeding index (PBI) were conducted. Saliva samples were collected for laboratory analysis performed by automated system Beckman Coulter AU480. Levels of uremic toxins (urea and creatinine) and secretory immunoglobulin A were determinated by spectrophotometric and immunoturbidimetric method, respectively. Oral swab and oral rinse method were used for microbiological examination. Results: The sociodemographic characteristics of the patients with CKD and healthy controls showed no significant differences. Predialysis subjects had significantly higher presence of oral mucosa changes and oral symptoms. Mean values of creatinine clearence were significantly lower in patients with oral mucosa pallor, uremic fetor, xerostomia and disguesia, compared to patients without listed symptoms. Predialysis patients showed significantly decreased salivary flow rate compared to both control groups and significantly increased pH values compared to healthy controls. Mean concentrations of salivary urea and creatinine were statistically different between the groups. Moderate positive correlation was determined between serum and salivary levels of urea and creatinine in predialysis patients and creatinine in hemodialysis patients. Statistical analysis showed no differences between groups in mean concentration of secretory immunoglobulin A. The rate of oral fungal colonisation was significantly higher in CKD patients with predominance of non-albicans Candida species. Conclusion: The results of the present study indicate the importance of determining the clinical characteristics of oral cavity in predialysis patients. Saliva flow rate, pH value and salivary concentration of uremic toxins could be reliable markers of kidney disease. Simple and non-invasive approach due to saliva sampling and reliability of laboratory test should contribute to a wider application of saliva as a competitive diagnostic fluid. Oral rinse technique is an accurate quantitative method for determining the rate of fungal colonization.</p>

The use of hair and oral fluid for drugs of abuse testing has increased over the last years. For this reason, the assurance that results provided using these matrices are reliable and error-free is needed. The objective of this thesis is to develop tools to assess the quality of results provided by laboratories analysing drugs of abuse in hair and in oral fluid and to evaluate the effect on the quality of results of different actions carried out. For this reason, intercomparison exercises have been organized and some studies have been performed to develop appropriate quality control materials. Regarding the analysis of drugs of abuse in hair, nine different intercomparison exercises have been organized. The evaluation of qualitative and quantitative results reported by laboratories together with the study of the methodology used, has led to know the quality of the results, to identify the sources of error and to know the corrective actions that should be developed. Concerning the quality control material, these exercises have enabled to know the influence on the results of the type of hair used to perform the analysis. Concerning the analysis of drugs of abuse in oral fluid, a method has been developed to identify and quantify 6-monoacetyl morphine, morphine, codeine, cocaine and benzoylecgonine in oral fluid samples. On the other hand, stability studies of the main drugs of abuse in oral fluid have been done to establish the optimal preparation, transport and storage conditions, and finally, two intercomparison exercises have been conducted to know the performance of analytical laboratories when analysing drugs of abuse in oral fluid and to know the stability of some drugs of abuse in two commercial collection devices. / La creixent utilització del cabell i del fluid oral per a l’anàlisi de drogues d’abús, ha portat a la necessitat d’assegurar que els resultats obtinguts utilitzant aquestes matrius són fiables i lliures d’error. L’objectiu d’aquesta tesi és desenvolupar eines que permetin avaluar la qualitat dels resultats dels laboratoris que analitzen drogues d’abús en cabell i en fluid oral i avaluar l’efecte de diferents accions sobre la qualitat final dels resultats. Per això s’han organitzat exercicis interlaboratori i s’han realitzat estudis per desenvolupar material d’assaig adequat. Pel que fa a l’anàlisi de drogues d’abús en cabell, s’han realitzat nou exercicis interlaboratori. A través de l’avaluació dels resultats qualitatius i quantitatius informats pels laboratoris i mitjançant l’estudi de la metodologia emprada, ha estat possible conèixer la qualitat dels resultats obtinguts, detectar les fonts d’error i conèixer quines mesures correctives caldria desenvolupar. Pel que fa al material d’assaig, aquests exercicis han permès conèixer la influència que té, en els resultats, el tipus de cabell del qual es parteix per realitzar l’anàlisi. Pel que fa a l’anàlisi de drogues d’abús en fluid oral, s’ha desenvolupat i validat un mètode analític que ha permès identificar i quantificar 6-monoacetil morfina, morfina, codeïna, cocaïna i benzoïlecgonina en mostres de fluid oral. Per altra banda, s’han realitzat estudis d’estabilitat de les principals drogues en fluid oral que han permès establir les condicions òptimes de preparació, transport i conservació del material d’assaig i, per últim, s’han realitzat dos exercicis interlaboratori que han permès conèixer la qualitat dels resultats analítics obtinguts pels laboratoris i també l’estabilitat de les drogues en dos dispositius de recollida comercials.