The aim of this work is to enhance the detection and classification pipelines of an artificial intelligence (AI)-based decision support system (DSS) for diagnosing acanthamoeba keratitis (AK), a vision-threatening disease. The images used are taken with the in vivo confocal microscopy (IVCM) technique, a complementary tool for clinical assessment of the cornea that requires manual human analysis to support diagnosis. The DSS facilitates automated image analysis and currently aids in diagnosing AK. However, the accuracy of AK detection needs improvements in order to use it in clinical practice. To address this challenge, we utilize image brightness processing through multiscale retinex (MSR), and develop a custom-built image processing pipeline with deep learning model and rule-based strategies. The proposed pipeline replaces two deep learning models in original DSS, resulting in an overall accuracy improvement of 10.23% on average. Additionally, our improved pipeline not only enhances the original system’s ability to aid AK diagnosis, but also provides a versatile set of functions that can be used to create pipelines for detecting similar keratitis diseases.

This thesis contributes to the development of dinoflagellate cysts as indicators of past environmental change in the Northeastern Pacific coastal ocean, and investigates past variations in sea-surface temperature, salinity and primary productivity encoded in dinoflagellate cyst sedimentary records from the Santa Barbara Basin (SBB, southern California) and Effingham Inlet (Vancouver Island, British Columbia) over the last millennium. The dinoflagellate cyst records extracted from the SBB and Effingham Inlet predominantly laminated sediments and analysed at sub-decadal resolutions, constitute some of the most detailed records of cyst-producing dinoflagellate populations in the world. A two year-long sediment trap study from the SBB documents the seasonality in dinoflagellate cyst production for the first time on the Pacific coast of the United States. The study shows that dinoflagellate cyst data can be used as indicators of changes in sea-surface temperature and primary productivity associated with seasonal upwelling in the SBB. In particular, several dinoflagellate cyst taxa such as Brigantedinium spp. and Lingulodinium machaerophorum are identified as indicators of “active upwelling” (typically occurring in spring and early summer) and “relaxed upwelling” conditions (fall and early winter) at the site, respectively. Analysis of a dinoflagellate cyst record from the SBB spanning the last ~260 years at biannual resolution documents the response of cyst-producing dinoflagellates to instrumentally-measured warming during the 20th century, and reveals decadal scale variations in primary productivity at the site that are coherent with phases of the Pacific Decadal Oscillation (PDO). The cyst assemblages are dominated by cysts produced by heterotrophic dinoflagellates (in particular Brigantedinium spp.), but the turn of the 20th century is marked by an abrupt increase in concentrations of L. machaerophorum and Spiniferites ramosus, two cyst taxa of autotrophic affinity. Their increasing abundances during the 20th century are interpreted to reflect warmer conditions and possibly stronger stratification during summer and fall. The dinoflagellate cyst data suggest a warming pulse in the early 1900s and provide further evidence that persistently warmer and/or more stratified conditions were established by the late 1920s. The dinoflagellate cyst record from Effingham Inlet, spanning the last millennium, is characterized by the proportionally equal contribution of cysts produced by autotrophic and heterotrophic dinoflagellates in most samples. The cyst data indicate variations in sea-surface temperature, salinity and primary productivity that are associated with local expressions of the Medieval Climate Anomaly (from the base of the record to ~1230), the Little Ice Age (from ~1230 to ~1900) and warming during the second half of the 20th century. Both dinoflagellate cyst records reveal that since the beginning (in the SBB) and mid-20th century (in Effingham Inlet), autotrophic dinoflagellates contribute to a greater portion of the primary production in the region, whereas heterotrophic dinoflagellates, as indicators of diatom populations, decline. Variability in the dinoflagellate cyst data is coherent at both sites and suggest a reduced expression of decadal scale variability associated with the PDO during the 19th century. / Graduate / 0416 / 0427 / mbringue@uvic.ca

During the Late Pliocene, between 3.3 Ma and 2.6 Ma, tectonic events changed ocean basin interactions against a background of shifting orbital forcing mechanisms and a global cooling trend. A climate system that had been locked in a stable, warm state gradually transformed into one typified by the high-amplitude glacial–interglacial fluctuations characteristic of the later Quaternary. The onset of Northern Hemisphere glaciations in the Late Pliocene marks an important step in this transition, due to the role of feedback mechanisms including ice albedo. A crucial factor in this Northern Hemisphere ice sheet expansion is the North Atlantic surface ocean circulation. To evaluate how they are linked, a ca. 200-kyr time slab spanning 2782-2520 ka (Late Pliocene–Early Pleistocene) was analyzed at millennial scale resolution from eastern North Atlantic Deep Sea Drilling Project (DSDP) Site 610 and Integrated Ocean Drilling Program (IODP) Site U1313. The causes of the Plio-Pleistocene climatic turnover are compared to that of the well-documented Marine Isotope Stage (MIS) M2 occurring in the Late Pliocene (3.3 Ma). MIS M2, a severe glacial event seen as a precursor to later Quaternary-style glaciations, was investigated from western North Atlantic DSDP Site 603. Utilizing a same-sample methodology, two paleoceanographic proxies were used: (1) dinocyst assemblages, and (2) foraminiferal geochemistry (δ18O and Mg/Ca). Dinocysts are proven tracers of sea-surface temperature (SST), salinity, nutrient supply, and sea ice cover, and are analyzed here to characterize the overlying water masses at the studied sites. Strong dinocyst assemblage fluctuations attest to variations in the influence of the North Atlantic Current (NAC). Using Mg/Ca ratios for the planktonic foraminifer Globigerina bulloides to determine absolute SSTs allows salinity changes to be reconstructed when combined with stable oxygen isotopes. This study shows a persistent Gulf Stream–NAC in the western North Atlantic during MIS M2, favoring a southern shift of the NAC over a shutdown of the thermohaline circulation. At the newly established Plio-Pleistocene boundary, a profound turnover in dinoflagellate cyst assemblages reveals a shift in ocean mode during MIS 104 (2.6 Ma). Three distinct dinocyst ecozones demonstrate this fundamental reorganization of the North Atlantic circulation.

Thesis (MMedSc )--University of Stellenbosch, 2011. / ENGLISH ABSTRACT: The present study investigated in particularly the specific nature of the supporting stromal layer located between the implanted human cyst and host murine tissue, which has yet to be reported. During an initial phase of this study, the particular light microscopic properties of the existing hematoxylin and eosin (H&E) stained experimental cyst was investigated, with regards to the presence or absence of specific morphological features, namely spongiosis, exocytosis, epithelial keratinization, epithelial thickness and hyperplasia, and the vascularity and fibrosis present in the stroma of these experimental sections. Subsequent analysis reported significant spongiosis, in addition to increased exocytosis of immune cells and epithelial keratinization in a number of cysts. Additionally, increased epithelial thickness and hyperplasia was reported in only 2 / 10 experimental tissues, whereas increased vascularity was observed in the stroma following analysis of H&E and Special staining, such as Verhoeff-von Gieson and Masson trichrome results. During the second phase of the study, immunohistochemical analysis with a particularly wide array of antibodies raised against specific human and mouse antigens had been applied. This involved automated immunohistochemical staining with mouse anti-human primary antibodies, in addition to manual staining with rabbit anti-mouse primary antibodies. Subsequent visualization was achieved by means of linking to biotinylated secondary antibodies, and Streptavidin-HRP incubation for standard visualization, followed by counterstaining with Hematoxylin. Maintained positive expression of cytokeratins 5, 13, and 14 was demonstrated in both control human vaginal mucosa and experimental cysts, whereas similar findings were not reported for cytokeratin 1, given the vast keratinization which was observed. Human collagen type IV and laminin of the basement membrane reported positive expression in 9 / 10 and 6 / 10 control human vaginal mucosa tissues respectively. In comparison, negative mouse collagen type IV and laminin was reported in most experimental cysts compared to positive staining in positive control mouse tissues. Immunohistochemical staining for human elastin, fibronectin, von Willebrand factor, and fibroblasts revealed maintained positive staining in all control human vaginal mucosa and experimental cysts. However, maintained expression of CD34 (endothelial marker), CD1a (langerhans cells), and human VEGFR-3 in experimental cysts was not demonstrated, compared to positive expression in control human vaginal mucosa. Subsequent analysis of murine antigens illustrated uniformly negative staining for mouse fibronectin, langerhans cells (CD207), and fibroblasts, in addition to negative staining in positive control mouse tissue sections. Furthermore, negative staining for mouse VEGFR-2 was reported in all experimental cysts; however strong positive staining of this marker in mouse kidney tissue had been reported. The findings of this study suggested that the exact nature of the stromal layer is of both human and murine origin. Furthermore, the tissue region located beneath the human vaginal epithelium is suggested to be of human nature, whereas the second distinct region located at the periphery of experimental cyst tissues, is suggested to be murine origin; however the findings of immunohistochemical analysis could not illustrate definitively the exact nature of the intermediate stromal layer, but could in fact demonstrate a mixture of human and murine tissue. / AFRIKAANSE OPSOMMING: Die huidige studie het die spesifieke molekulêre en histologiese eienskappe van die stromale laag geleë tussen menslike sist- en muis velweefsel bestudeer, wat tans nog nie bekend is nie. Gedurende die eerste fase van hierdie studie is die besondere lig-mikroskopiese eienskappe van die bestaande hematoksilien en eosien (H&E) eksperimentele siste bestudeer, met betrekking tot die aan- of afwesigheid van spesifieke morfologiese eienskappe, naamlik spongiose, eksositose van immuunselle, epiteel keratinisasie, epiteel dikte en hiperplasie, en laastens die stromale vaskulariteit en fibrose. Gevolglike analise het daarop gedui dat beduidende spongiose, eksositose en epiteel keratinisasie gevind word in die eksperimentele siste in vergelyking met kontrole vaginal weefsel. Hierteenoor is die verdikking van die epiteel en hiperplasie in slegs 2 / 10 eksperimentele siste gevind, terwyl vermeerderde vaskulariteit aangedui is na gevolglike H&E en spesiale (soos byvoorbeeld Verhoeff-von Gieson en Masson trichrome) kleuringsresultate. Die tweede fase van die studie het die immunokleuring met verskeie mens- en muis spesifieke antiliggame behels, waarby die uitdrukking van verskeie mens antigene vergelyk is met dié van muis. As sulks is ge-automatiseerde immunohistochemie toegepas met muis primêre antiliggame, tesame met fisiese kleuring met konyn primêre antiliggame toegepas. Gevolglike visualisasie is aangedui deur middel van binding met sekondêre antiliggaam en Streptavidin- HRP, gevolg deur teenkleuring met Hematoksilien. Algehele behoud van positiewe uitdrukking van sitokeratien 5, 13, en 14 is bevind, terwyl sitokeratien 1 uitdrukking nie daarwerklik vergelykbaar is met dié van kontrole mens vaginale weefsel nie. Die uitdrukking van mens kollageen IV en laminien van die basaal membraan is verder bestudeer, en het egter positiewe kleuring in 9 / 10 en 6 / 10 van kontrole mens vaginale mukosa aangedui. In vergelykking hiermee kon die huidige bevindings egter net positiewe kleuring in 4 / 10 en 3 / 10 eksperimentele siste vir kollageen IV en laminien onderskeidelik, illustreer. Immunohistochemiese analise van menslike elastien, fibronektien, von Willebrand (vW) faktor en fibroblaste het op deurgaans positiewe uitdrukking van hierdie merkers aangedui in beide eksperimentele en kontrole menslike weefsel. In teenstelling hiermee is volgehoue uitdrukking van CD34 (endoteel merker), CD1a (Langerhans sel merker) en mens VEGFR-3 in ekperimentele siste egter nie illustreerbaar nie, in vergelykking met deurgaans positiewe uitdrukking van hierdie antigene in kontrole mens vaginale mukosa. In opvolging is deurgaans negatiewe uitdrukking van muis fibronektien, langerhans sel (CD207) en fibroblaste bevestig, terwyl negatiewe kleuring ook deurgaans in positiwe kontrole muis weefsel, bekom deur die disseksie van ‘n naakte muis, gevind is. Verder is ook negatiewe kleuring vir VEGFR-2 in alle eksperimentele siste gevind, terwyl egter sterk positiewe kleuring in muis nierweefsel as positiewe weefsel gevind is. Die resultate van die huidige studie het daarop gedui dat die stromale laag onderliggend tot mens vaginale epiteel van menslike oorsprong is, terwyl die periferale stroma onderliggend tot muis velweefsel, ongetwyfeld van muis oorsprong is. Laastens kon die spesifieke oorsprong van die tussenliggende stroma nie aangedui word nie, maar dat dit moontlik uit beide menslike- en muisweefsel bestaan.

O queratocisto odontogênico e o ameloblastoma sólido convencional/multicístico são lesões localmente agressivas que se desenvolvem nos ossos maxilares e que possuem uma alta taxa de recidiva se não forem tratadas apropriadamente. Portanto, o principal objetivo da terapia cirúrgica é a erradicação da lesão, diminuindo a possibilidade de recidivas pós-operatórias. Essas cirurgias geralmente causam um impacto negativo na estética e função do sistema estomatognático, sendo fundamental um esforço em busca de procedimentos que não causem um dano anatômico tão grande aos pacientes e que sejam suficientemente seguros no controle de recidivas. Contudo, muitas vezes a escolha da técnica operatória se apoia em conhecimentos de relatos/séries de casos e de experiêcias individuais, havendo uma carência de dados que consolidem determinado protocolo de tratamento, gerando dúvidas na tomada de decisões clínicas. O objetivo do presente estudo é fornecer suporte científico para auxiliar na tomada de decisão sobre a implementação de determinadas técnicas cirúrgicas conservadoras no tratamento do queratocisto e do ameloblastoma sólido convencional/multicístico por meio de quatro artigos científicos cuja elaboração reuniu questões relevantes na prática clínica. / Odontogenic keratocyst and solid/multicystic ameloblastoma are locally aggressive lesions of the jaws that have a high recurrence rate if not properly treated. Therefore, the main objective of the surgical treatment is the eradication of the lesion and the control of postoperative recurrences. These surgeries generally have a negative impact on the function and aesthetics of the stomatognathic system and efforts are necessary to conduct procedures that do not cause an extensive anatomical damage to the patients and that are sufficiently safe in the control of recurrence rate. However, the decision of a surgical technique is often based on results from case reports/case series, lacking of data that consolidates determinate treatment protocol, leading to doubts in the clinical decision. The aim of the present study is to provide scientific support to aid in the decision-making on the implementation of certain conservative surgical techniques in the treatment of the keratocyst and solid ameloblastoma through the presentation of four scientific articles which brought relevant topics in clinical practice.

Knowing the taxonomy of unicellular dinoflagellate cysts is important in reconstructing paleoenvironments, paleoclimatology and paleobathymetry. Different dinoflagellates tend to prefer different depths, salinity levels and climate. Species of dinocysts only occur during a certain interval in the geological record and are therefore an important tool for dating sediments. Biostratigraphic research uses age determination of sediments depending on first and last occurrences of different species, or co-occurrences of several species. However, it can be rather time consuming and difficult to identify dinoflagellate cysts for people that are not experts in this field. In an attempt to resolve this, a database was created for 145 genera of dinoflagellate cysts existing during Palaeocene and Eocene in the online accessible program Xper3. Morphological criteria or “descriptors” were added along with describing “states”. The number ofmorphological descriptors chosen were 13, with 3 to 15 states for each descriptor.Each genus was described in a matrix and was given one or more unique states for each descriptor, which were defined from the literature by original morphological descriptions of dinocyst genera and photographs of holotypes. A morphological identification key was automatically created in Xper3 from this database, which enabled identification of genera by choosing unique states that were visible for the dinocysts being analysed.A test using photographs with different levels of preservation of dinoflagellate cysts of Eocene age of Aquitaine basin was undertaken in order to assess the reliability of the morphological identification key. The aim was also to see which morphological criteria were more common and how many descriptors and states were needed to reach an end-result of 5 remaining genera, including the correct genus.Errors that were caused in the identification key were immediately reviewed and re-tested. In 38 of 43 tries, a maximum of five genera were remaining, with the correct genus included. This confirmed that the identification key worked relatively well.Another test demonstrated how the identification key worked for identifying dinocysts with an optical microscope in unknown samples, which are photographed and published in the present report, along with data showing how many descriptors and states were used, remaining genera and which states were chosen for each genus.In general, only 1-8 descriptors were necessary and 5 the most common number used. The morphological character (state) “type of ornamentation” was the most widely used followed by “distribution of ornamentation”, “shape of the cyst” and “size of the ornamentation”. However, the archeopyle (excystment opening), was not always visible in the dinocysts, but is still considered a key morphological descriptor. / Dinoflagellater är en typ av mikroskopiska plankton som kan bilda så kallade cystor eller vilosporer för att skydda sig vid kritiska miljöförändringar. Dessa cystor kan ligga vilandes i sediment under en längre tid. Det är också dessa cystor som bevaras som fossil. Eftersom det finns en sådan hög mångfald av dinoflagellatcystor i fossila sediment, liksom att de mikroskopiska, existerar över stora delar av jorden samt att de har en snabb evolution, så används de ofta till att datera sediment inom biostratigrafi.För att kunna datera sediment med hjälp av dinoflagellatcystor, krävs det att man kan identifiera de olika arterna och veta i vilken tidsålder de existerade. Att klassificera dinoflagellatcystor kan därför vara mycket svårt och ta en hel del tid om man inte är någon expert. I denna studie har därför en databas och identifieringsnyckel skapats i ett onlinebaserat dataprogram kallat Xper3. Xper3 är primärt riktat till att hjälpa personer med grundläggande kunskaper i att beskriva utseendemässiga karaktärer hos olika typer av mikrofossil, inklusive dinoflagellatcystor.I denna identifieringsnyckel använde jag mig av 145 olika dinoflagellatsläkten vilka alla existerade någon gång under tidig Paleogen, dvs hela Paleocen och/eller Eocen (66–34 miljoner år sedan). Dessutom användes 13 olika morfologiska beskrivningar ”descriptors” med 3–15 olika svar ”states” per beskrivning.Målet är att få en identifieringsnyckel som unikt kan identifiera olika typer av dinoflagellatcystor på genusnivå, med ett förslag på maximalt fem återstående släkten (genera). Studien undersökte även vilka morfologiska beskrivningar som är mest användbara och hur många olika typer av beskrivningar som behövs för att analysera dinoflagellatcystor.För att testa om identifieringsnyckeln fungerade, användes olika typer av kända och okända dino-flagellatcystor i ett så kallat ”Test 1: Kända dinoflagellatcystor” och ”Test 2: Okända dinoflagellat-cystor.I det första testet, ”Test 1”, grupperades kända dinoflagellatcystor i olika typer av bevarande tillstånd så som ”välbevarade”, ”tveksamt bevarande” och ”väldigt dåligt bevarade” dinoflagellat-cystor. Detta test förde statistik på hur många släkten av förslag som kom upp i slutändan och vilka morfologiska beskrivningar som användes mest och hur många olika beskrivningar som behövdes.Det andra testet, ”Test 2”, undersökte hur denna identifieringsnyckel fungerade att analysera dinoflagellatcystor från okända prover under mikroskop. Fotograferade dinoflagellatcystor presente-rades i denna rapport tillsammans med detaljer på hur de hade identifierats.I de flesta fallen, kunde identifieringsnyckeln ge ett förslag på maximalt fem återstående släkten av dinoflagellatcystor, där det korrekta släktet var ett utav förslaget, vilket tyder på att identifierings-nyckeln fungerar. Den mest använda morfologiska beskrivningen var ”typ av ornament”, och antal morfologiska beskrivningar som behövdes var mellan ett till åtta, där fem var det antal beskrivningar som mest användes i att identifiera dinoflagellatcystor i identifieringsnyckeln.

Entre as lesões odontogênicas não mineralizadas, o ameloblastoma, o tumor odontogênico queratocístico e o cisto dentígero podem apresentar-se indistinguível na imagem radiográfica. A dificuldade na interpretação da imagem e elaboração do diagnóstico destas lesões é tema abordado em vários estudos que utilizaram exames de imagem avançados, como ressonância magnética e tomografia computadorizada, para definir como podem colaborar na diferenciação, na avaliação da extensão e no planejamento cirúrgico das mesmas. Com a recente introdução da Tomografia Computadorizada de Feixe Cônico (TCFC) como recurso auxiliar de diagnóstico, o presente trabalho foi elaborado tendo em vista descobrir como o diagnóstico pode ou não ser influenciado por essa nova tecnologia por meio da comparação das hipóteses de diagnóstico obtidas em imagens das radiografias panorâmicas e em TCFC, em casos de ameloblastoma, tumor odontogênico queratocístico e cisto dentígero; e também definir as características observadas nas imagens de TCFC que podem auxiliar no diagnóstico diferencial. Foram selecionados 5 casos de cada lesão. As radiografias panorâmicas e os exames de TCFC foram analisados por 15 cirurgiões-dentistas - 4 especialistas em Radiologia, 4 Mestres e 7 Doutores em Estomatologia e Radiologia - para a formulação das hipóteses de diagnóstico. Na primeira etapa foram analisadas as radiografias panorâmicas e na segunda, após um intervalo de pelo menos 30 dias, os exames de TCFC. Dois examinadores, capacitados e calibrados, fizeram a análise e mensuração das características qualitativas e quantitativas das lesões observadas nos exames de TCFC, utilizando o programa i-CAT® Vision. Os resultados demonstraram que não houve diferença estatisticamente significante na média total dos acertos de diagnóstico entre radiografia panorâmica e TCFC, porém houve diferença significante na média de acerto no diagnóstico de ameloblastoma com o uso de TCFC (77,33% ± 21,20%) em comparação à radiografia panorâmica (61,33% ± 23,26%). Os mestres (85,00% ± 19,15%) e doutores (65,75% ± 9,76%) obtiveram uma média de acerto maior no diagnóstico de tumor odontogênico queratocístico com o uso da radiografia panorâmica em relação aos especialistas (35,00% ± 19,15%), com diferença significante. A análise das características quantitativas observadas nas imagens de TCFC revelou que o ameloblastoma mostrou-se maior em tamanho e em expansão comparado ao tumor odontogênico queratocístico e ao cisto dentígero, com diferença estatisticamente significante. De todas as características qualitativas avaliadas, apenas o número de lóculos da lesão (multilocular ou unilocular) mostrou diferença significante, o ameloblastoma apresentou uma maior incidência do aspecto multiloculado comparado ao tumor odontogênico queratocístico e ao cisto dentígero. Não houve diferenças significantes entre as características quantitativas e qualitativas do tumor odontogênico queratocístico e do cisto dentígero. / Among the non-mineralized odontogenic lesions, ameloblastomas, odontogenic keratocystic tumors and dentigerous cysts may present as radiographically indistinguishable. The difficulty in radiographic diagnosis of these lesions is the subject addressed in several studies using advanced imaging, such as magnetic resonance imaging and computed tomography, to define how these technologies can help in differentiating among these lesions, in assessing the extent and surgical planning. Cone-beam computed tomography (CBCT) is a recent diagnostic aid resource and this study was prepared in order to determine how the diagnosis may or may not be influenced by this new technology, comparing the diagnostic hypotheses obtained using images of panoramic radiographs and CBCT in cases of ameloblastoma, odontogenic keratocystic tumor, and dentigerous cyst - and to define the features observed in CBCT images that can help the differential diagnosis. Five cases were selected for each lesion. Panoramic radiographs and CBCT scans were analyzed by 15 dentists - 4 specialists in Radiology, 4 masters and 7 PhD in Stomatology and Radiology - for the formulation of the diagnostic hypotheses. In the first stage the panoramic radiographs were analyzed, and in the second, after an interval of at least 30 days, the CBCT images. Two observers, trained and calibrated, performed the analyses and measurement of qualitative and quantitative features of the lesions evaluated in the CBCT images using the i-CAT® Vision program. The results showed that there was no statistically significant difference in correct diagnostic average between panoramic radiography and CBCT, but there was a significant difference in correct diagnostic average in the diagnosis of ameloblastoma using CBCT (77.33% ± 21.20%) compared to panoramic radiography (61.33% ± 23.26%). Masters (85.00% ± 19.15%) and PhD-level observers (65.75% ± 9.76%) had greater correct diagnostic average in the diagnosis of odontogenic keratocystic tumor using panoramic radiograph compared to specialists (35.00% ± 19.15%), with a significant difference. The analysis of quantitative features observed in the CBCT images revealed that the ameloblastomas were greater in size and expansion compared to the odontogenic keratocystic tumors and the dentigerous cysts, with statistically significant difference. Of all the qualitative features evaluated, only the number of loci of the lesion (multilocular or unilocular) showed significant differences; ameloblastomas showed a higher incidence of multiloculated aspects compared to odontogenic keratocystic tumors and dentigerous cysts. There were no significant differences between quantitative and qualitative features of odontogenic keratocystic tumors and dentigerous cysts.

Made available in DSpace on 2017-07-10T14:57:30Z (GMT). No. of bitstreams: 1 Dissertacao_Renata Zoraida_ Rizental Delgado.pdf: 2641394 bytes, checksum: c9a23494ed97bde58dcc144b147a740f (MD5) Previous issue date: 2015-02-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Introduction: Gorlin syndrome (GS) also known as Nevoid Basal Cell Carcinoma Syndrome, is a rare disease resulting from mutations in Patched-1 gene and characterized by triad of disorders comprising multiple basal cell carcinomas, numerous keratocystic odontogenic tumors (KOT) and skeletal abnormalities. About 90% of patients develop KOTs in gnathic bones, preferably in the posterior mandible, presenting radiographically as radiolucent lesions uni- or multilocular. Studies suggests that KOTs associated with GS (KOTGSs) have distinct and more aggressive behavior compared to those who developing sporadically (KOTSPs). Objectives: This study aimed at comparing clinical, histopathological and immunohistochemical features of KOTSPs and KOTGSs from different institutions in Brazil and abroad, and understand the role of proteins associated with proliferation/cell cycle (p53, p63 and Ki-67), the alpha-smooth muscle actin (α-SMA) and syndecan-1 (CD138) in an attempt to associate the expression with biological behavior of KOTs. Methodology: Were previously reviewed and selected 30 KOTGSs and 8 KOTSPs, in which were performed qualitative and semi-quantitative analysis histopathological and immunohistochemical to p53, p63, Ki-67, SMA and CD138. Results: 30 cases of KOTGSs were obtained from 12 patients with GS, 5 females (41.66%) and 7 men (58.33%); whereas in group of KOTSPs, 5 cases (62.50%) was in females and 3 males (37.50%). About 58.33% of patients with GS had more than one lesion throughout life. The average age of individuals with GS was 14.66 ± 16.81 years, while in patients with KOTSPs was 41 ± 39.59. The predominant radiographic pattern was radiolucent unilocular, preferably affecting the posterior mandible. Histopathological features in both groups was analyzed, and the most frequent was pleomorphism in KOTGSs. In addition, there was increased expression of p53 and p63 in KOTGSs and similar expression of SMA and Ki-67 between groups. It was also observed that there was a lower reactivity for CD138 in the basal epithelial layer of KOTGSs and stromal expression of CD138 was similar between the groups. Conclusions: Agressiveness of KOTGSs can be explained by increased cellular pleomorphism rate and expression of p53 and p63 and tendency to loss of syndecan-1 expression compared to KOTSPs. / Introdução: A Síndrome de Gorlin (SG) também conhecida como Síndrome do Carcinoma Nevóide Basocelular, é uma doença rara resultante de mutações no gene Patched-1 e caracterizada por uma tríade de alterações que inclui múltiplos carcinomas basocelulares, numerosos tumores odontogênicos queratocísticos (TOQs) e anormalidades esqueléticas. Cerca de 90% dos portadores desenvolvem TOQs no interior dos ossos gnáticos, preferencialmente na região posterior da mandíbula, apresentando-se radiograficamente como lesões radiolúcidas uni ou multiloculares. Estudos têm sugerido que os TOQs associados à SG (TOQSG) apresentam comportamento distinto e mais agressivo em comparação àqueles que desenvolvem-se de modo esporádico (TOQEs). Objetivos: O presente estudo teve como objetivo analisar comparativamente os aspectos clínicos, histopatológicos e imuno-histoquímicos de TOQEs e TOQSGs oriundos de diferentes instituições do Brasil e do exterior, além de entender o papel de proteínas associadas com proliferação/ciclo celular (p53, p63 e Ki-67), da actina de músculo liso-alfa (α-SMA) e da proteína sindecano-1 (CD138), na tentativa de associar a expressão das mesmas com o comportamento biológico dos TOQs. Metodologia: Foram previamente revisados e selecionados 30 TOQSGs e 8 TOQEs, nos quais foram realizadas análises qualitativas e semi-quantitativas histopatológicas e imuno-histoquímicas para p53, p63, Ki-67, AML e CD138. Resultados: Trinta casos de TOQSGs foram obtidos de 12 pacientes com SG, 5 do gênero feminino (41,66%) e 7 do gênero masculino (58,33%); ao passo que no grupo dos TOQEs, 5 casos (62,50%) acometeram o gênero feminino e 3 o masculino (37,50%). Cerca de 58,33% dos pacientes com SG apresentaram mais de uma lesão ao longo da vida. A média de idade dos indivíduos com SG foi de 16,81 anos ± 14,66 enquanto dos portadores de TOEs foi de 41 anos ± 39,59. O padrão radiográfico predominante foi o radiolúcido unilocular, afetando preferencialmente a região posterior de mandíbula. Foram avaliadas as características histopatológicas de ambos os grupos de lesões, sendo o pleomorismo celular mais frequente nos TOQSGs. Além disso, observou-se maior expressão de p53 e p63 nos TOQSG e expressão similar de AML e Ki-67 entre os grupos. Observou-se ainda que houve menor reatividade para CD138 no estrato epitelial basal dos TOQSGs e a expressão estromal de CD138 foi similar entre os grupos analisados. Conclusões: A maior agressividade dos TOQSGs pode ser explicada pela maior taxa de pleomorfismo celular, maior expressão de p53 e p63 e tendência à perda de expressão de sindecano-1 quando comparados aos TOQEs.

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Die Toxoplasmose gehört zu den bedeutsamsten Zoonosen. Sie wird durch den weltweit verbreiteten Erreger T. gondii ausgelöst und verläuft bei immunkompetenten Menschen meist subklinisch. Dennoch können Rezidivierungen und schwere Krankheitsverläufe auftreten. Besonders gefährdete Personen sind Immunsupprimierte und seronegative Schwangere. T. gondii kann alle warmblütigen Lebewesen infizieren. In der Folge kommt es zur Bildung von Gewebezysten in Muskulatur und Organen. Daten zum Vorkommen und zur Verbreitung des Erregers bei deutschen Schlachtputen liegen nicht vor und sollten im Rahmen dieser Arbeit gewonnen werden. Das hohe Infektionsrisiko bei oraler Aufnahme von rohem oder nicht vollständig durchgegartem zystenhaltigem Fleisch ist bekannt. Exakte Erkenntnisse zur Tenazität der T.-gondii-Gewebezysten gegenüber dem Einfluss von Salz, pH-Wert und Wurstreifungsprozessen liegen jedoch nicht vor. Da sowohl Schweine- als auch Putenfleisch häufig zur Herstellung von Fleischerzeugnissen wie z. B. Rohwürsten und Rohschinken eingesetzt wird, sollte die Persistenz der Gewebezysten unter dem Einfluss verschiedener Salzgehalte und pH-Werte sowie kombiniert im Rahmen der Wurstreifungsprozesse untersucht werden. Ziel ist eine genauere Beurteilung des Infektionsrisikos beim Verzehr kurz gereifter Rohwürste (Mettwurst) und anderer nicht erhitzter Fleischzeugnisse. Untersuchungen: 1. Es wurden 1913 Blutserumproben von Schlachtputen aus 14 Mastbetrieben in fünf Bundesländern mittels kinetischem ELISA auf das Vorkommen von IgG-Antikörpern gegen T. gondii untersucht. 2. In den In-vitro-Studien wurde der Einfluss von NaCl bzw. Nitritpökelsalz (NPS) in wurstrelevanten Konzentrationen (2,0, 2,5 3,0 %) und des pH-Wertes (5, 6, 7) auf die Gewebezysten in Mäusemuskulatur und -hirn untersucht. Die Überprüfung der Infektiosität der exponierten Gewebezysten erfolgte mittels Maus-Bioassay. 3. Putenrohwurstbrät (2,0 % NaCl oder NPS, mit und ohne Starterkultur) wurde mit zystenhaltiger Mäusemuskulatur beimpft. Während der Wurstreifung wurde die Infektiosität der Zysten im Maus-Bioassay untersucht. Ergebnisse:  Die T.-gondii-Gesamtseroprävalenz in deutschen Schlachtputenbeständen betrug 18,4 %. Dieser Wert ist überraschend hoch. Es handelt sich um die ersten in Deutschland durchgeführten Erhebungen. Jahreszeitliche Schwankungen wurden beobachtet, wobei in den Wintermonaten gemästete Putenherden eine höhere Prävalenz zeigten als die im Sommer gehaltenen Tiere.  Gewebezysten besitzen eine gewisse pH-Wert-Toleranz. Sie blieben bei pH-Werten von 5 bis 7 bis zu 26 Tage infektiös.  Gewebezysten sind gegenüber dem Einfluss von Salz empfindlich. Bei NaCl Konzentrationen von 2,0 % blieben sie bis zu acht Tagen infektiös, bei  2,5 % maximal ein Tag.  Gegenüber NPS sind die Zysten offenbar empfindlicher als gegenüber NaCl; sie blieben bei 2,0 % NPS-Konzentration vier Tage infektiös.  Bei Kombination der Effekte von Salz und pH-Wert verkürzt sich die Infektionsfähigkeit der Zysten in der Rohwurstreifung auf ca. 24 Stunden. Schlussfolgerungen: Die unerwartet hohen Seroprävalenzen weisen daraufhin, dass Putenfleisch ein Infektionsrisiko für Toxoplasmose darstellen kann. Weiterführende Untersuchungen zum Vorkommen des Erregers in Schlachtputen sind notwendig. Tendenziell zeigen die Ergebnisse der Tenazitätsuntersuchungen einen raschen Verlust der Infektiosität des Erregers, so dass in lang gereiften Rohwürsten oder in herkömmlich produzierten kurz gereiften Rohwürsten (NPS > 2,5 %) nicht mit einem Vorkommen von infektiösen Zysten zu rechnen ist. Eine Ausnahme könnten allerdings salzreduzierte (NaCl  2,0 %), ohne NPS hergestellte oder sehr kurz gereifte Rohwürste darstellen, hierbei kann ein Infektionsrisiko nicht vollständig ausgeschlossen werden. Zur Beurteilung von Rohschinken sind weitere Untersuchungen notwendig.