El adenocarcinoma ductal pancreático (ADP) constituye aproximadamente el 85% de los tumores del páncreas. Es el tumor en el que menos avances clínicos se han producido en los últimos 50 años y sigue teniendo un pronóstico muy malo. Por ello, es necesaria la identificación de factores de riesgo y la caracterización del proceso de progresión tumoral, así como la identificación de nuevas dianas terapéuticas. En esta Tesis Doctoral estudiamos el estado de las vías de señalización celular en una serie de ADP convencionales, así como en ductos de pancreatitis crónicas (PC) y las lesiones precursoras asociadas (PanIN). El objetivo principal del presente trabajo es el estudio inmunohistoquímico y molecular de diversos factores implicados en las vías de señalización celular y de transición epitelio-mesénquima (PI-3K-AKT-mTOR, Ras-Raf-MAPK, Hippo, p53, N-cadherina y p16), con el objetivo de poder definir un modelo de progresión desde lesiones de PanIN en PC a ADP infiltrante , así como demostrar la relación de los niveles de expresión de dichos factores con una serie de parámetros clínico-patológicos con valor pronóstico, estado mutacional y supervivencias, en el ADP. Los resultados del estudio de nuestra serie validan, por una parte, los criterios establecidos como factores con valor pronóstico: tamaño tumoral, margen quirúrgico afecto (R1), invasión vascular, invasión linfática, invasión perineural, metástasi hepáticas y estadios avanzados. En relación al modelo de progresión PanIN-ADP, demostramos que existe un aumento en la expresión de la gran mayoría de proteínas estudiadas desde su expresión en ductos de PC y la expresión en ADP. Asimismo, porponemos un valor de Ki67 ≥ a 20% como biomarcador útil a la hora de discernir entre proliferación ductular reactiva o ADP bien diferenciado en las biopsias pequeñas o en los márgenes quirúrgicos de piezas de resección. En relación al estudio genético de RAS, confirmamos que ningún caso presenta mutación para NRAS, a diferencia del 89% de los ADP, que presentan mutaciones de KRAS, y que la presencia de dicha mutación implica peor pronóstico. En cuanto a los factores de señalización celular en ADP, se confirma que la expresión de algunas proteínas (p4E-BP1, eIF4E y YAP1) se asocia a factores de mal pronóstico y peor supervivencia. No evidenciamos en el estudio relación significativa entre la expresión de las proteínas estudiadas con parámetros clínico-patológicos relacionados con estroma peritumoral (p16, desmoplasia, inflamación). Nuestros resultados concluyen que el estudio de los factores de las vías de señalización celular complementan el estudio anatomopatológico de los ADP. La determinación de Ki67 con un valor superior a 20% permite discernir entre una proliferación ductular reactiva/metaplasia acinar ductal y ADP bien diferenciado y lo proponemos como marcador a la hora de valorar margen quirúrgico de piezas de resección y biopsias pequeñas. La sobreexpresión de YAP1 nuclear, eIF4E citoplasmático y p4E-BP1 nuclear definen un perfil molecular de ADP de peor pronóstico. Finalmente, resaltar la relevancia de caracterizar factores asociados al microambiente que rodea al ADP e influyan en el comportamiento biológico de los mismos. El grupo de tumores sin mutaciones de KRAS y NRAS es muy interesante de cara a identificar otras eventuales alteraciones moleculares, con incidencia en el desarrollo de los ADP. La intención en un futuro es ampliar la serie para conseguir resultados significativos que permitan obtener más conclusiones. / Pancreatic ductal adenocarcinoma (ADP) constitutes approximately 85% of pancreatic tumors. It is the tumor in which fewer clinical developments have occurred in the last 50 years and continues to have a very poor prognosis. Therefore, the identification of risk factors and characterization of tumor progression, as well as the identification of new therapeutic targets is necessary. In this Thesis we studied the state of the cell signaling pathways in a number of conventional ADP as well as ducts of chronic pancreatitis (CP) and the associated precursor lesions (PanIN). The main objective of this paper is the study of various factors involved in cell signaling pathways and epitelio-mesenchymal transition (PI-3K-AKT-mTOR, Ras-Raf-MAPK, Hippo, p53, N-cadherin and p16) by immunohistochemical and molecular study, with the aim to define a model of progression from precursor lesions of PanIN in PC to infiltrating ADP; and to demonstrate the relationship of the expression levels of these factors with a series of pathological parameters with prognostic value, mutational state and survival in the ADP. The survey results of our series validate the established prognostic criteria factors: tumor size, surgical margin affection (R1), vascular invasion, lymphatic invasion, perineural invasion, progression to liver metastasis and advanced stages. Regarding the progression model PanIN-ADP show an increased expression of the vast majority of proteins studied since its expression in ducts of PC to ADP expression. Also, we propose a value of Ki67 ≥ 20% as useful biomarker to discern between a benign reactive ductular proliferation/acinar ductal metaplasia and a well differentiated ADP in small biopsies or in surgical margins of resected specimens. In relation to genetic study of RAS, we confirm that no event has mutation to NRAS, unlike the 89% of ADP showing KRAS mutations, and the presence of said mutation confers decreased survival. As for cell signaling factors in ADP, it is confirmed that the expression of some proteins (p4E-BP1, eIF4E and YAP1) factors associated with poor prognosis and shorter survival. No we show in the study significant relationship between the expression of the proteins with pathological parameters related to peritumoral stroma (p16, desmoplasia, inflammation). Our results conclude that the study of the factors of cellular signaling pathways complement the pathological report of the ADP. Ki67 determining a value greater than 20% can discern between a benign reactive ductular proliferation and a well differentiated ADP and propose it as a marker in assessing surgical resection margin and small biopsies. Overexpression of YAP1 nuclear, eIF4E cytoplasmic and p4E-BP1 nuclear define a molecular profile of ADP with worse prognosis. Finally, highlight the importance of characterizing factors associated with ADP microenvironment and their influence on biological behavior. The group of tumors without KRAS mutations and NRAS is very interesting in order to identify other molecular alterations, with impact on the development of ADP. The intention in the future is to expand the series to achieve significant results that allow more conclusions.

The recent finding that MYC-driven cancers are sensitive to inhibition of the DNA damage response (DDR) pathway, prompted us to investigate the role of DDR pathway as therapeutic target in diffuse large B-cell lymphoma (DLBCL), which frequently overexpresses the MYC oncogene. In a preliminary immunohistochemical study conducted on 99 consecutive DLBCL patients, we found that about half of DLBCLs showed constitutive expression of the phosphorylated forms of checkpoint kinases (CHK) and CDC25c, markers of DDR activation, and of phosphorylated histone H2AX (γH2AX), marker of DNA damage and genomic instability. Constitutive γH2AX expression correlated with c-MYC levels and DDR activation, and defined a subset of tumors characterised by poor outcome. Next, we used the CHK inhibitor PF-0477736 as a tool to investigate whether the inhibition of the DDR pathway might represent a novel therapeutic approach in DLBCL. Submicromolar concentrations of PF-0477736 hindered proliferation in DLBCL cell lines with activated DDR pathway. These results were fully recapitulated with a different CHK inhibitor (AZD-7762). Inhibition of checkpoint kinases induced rapid DNA damage accumulation and apoptosis in DLBCL cell lines and primary cells. These data suggest that pharmacologic inhibition of DDR through targeting of CHK kinases may represent a novel therapeutic strategy in DLBCL. The second part of this work is the clinical, molecular and functional description of a paradigmatic case of primary refractory Burkitt lymphoma characterized by spatial intratumor heterogeneity for the TP53 mutational status, high expression levels of genomic instability and DDR activation markers, primary resistance to chemotherapy and exquisite sensitivity to DDR inhibitors.

La PKCε e la PKCδ, chinasi ubiquitariamente distribuite e ad azione pleiotropica, sono implicate del differenziamento, sopravvivenza e proliferazione cellulare. Esse sono coinvolte nel processo differenziativo delle cellule staminali ematopoietiche e in fenomeni patologici associati al compartimento sanguigno. In questa tesi sono presentati i risultati riguardanti lo studio in vitro del ruolo di PKCε e PKCδ nel contesto del differenziamento megacariocitario, in particolare si caratterizza l’espressione e la funzione di queste chinasi nel modello umano e nel modello murino di Megacariocitopoiesi, normale e patologica. Confrontando le cinetiche dei due modelli presi in analisi nello studio è stato possibile osservare come in entrambi PKCε e PKCδ dimostrino avere una chiara e specifica modulazione nel progredire del processo differenziativo. Questi dati, se confrontati, permettono di affermare che PKCε e PKCδ presentano un pattern di espressione opposto e, nel modello umano rispetto a quello murino, reciproco: nell’uomo i livelli di PKCε devono essere down-modulati, mentre nel topo, al contrario, i livelli della chinasi risultano up-modulati durante lo stesso processo. Analogamente, le CD34+ in differenziazione presentano una costante e maggiore espressione di PKCδ durante la maturazione MK, mentre nel modello murino tale proteina risulta down-modulata nella fase più tardiva di formazione della piastrina. Le chinasi mostrano in oltre di agire, nei due modelli, attraverso pathways distinti e cioè RhoA nel topo e Bcl-xL nell’uomo. È stato inoltre verificato che l’aberrante differenziamento MK osservato nella mielofibrosi primaria (PMF), è associato a difetti di espressione di PKCε e di Bcl-xL e che una forzata down-modulazione di PKCε porta ad un ripristino di un normale livello di espressione di Bcl-xL così come della popolazione di megacariociti formanti propiastrine. I dati ottenuti indicano quindi che PKCε e PKCδ svolgono un ruolo importante nel corretto differenziamento MK e che PKCε potrebbe essere un potenziale nuovo target terapeutico nelle PMF. / Protein kinases C (PKC) are known to be ubiquitously distributed and to have pleiotropic effects. Isoforms epsilon (PKCε) and delta (PKCδ) are involved in the regulation of cell growth, survival and differentiation; in particular, they have been also investigated for their role in the hematopoiesis and in aberrant processes of differentiation along the erythroid and megakaryocytic lineages. In this PhD thesis, the results of an in vitro study about the role of these two kinases in models of megakaryocytic (MK) differentiation, both normal and pathological, are presented. The observations about PKCε and PKCδ kinetics show how these proteins have a specific modulation during the MK differentiation that results in an opposite pattern of expression and, in the murine model if compared with the human model, also a reciprocal one. In particular, in human megakaryocytopoiesis, PKCε results down-modulated, whereas in mouse its levels increase. Instead, PKCδ shows a high and steady expression in maturing CD34+ MK committed, but it is strongly down-modulated during the latest phases of platelet maturation in the murine model. The study also elucidates the different pathways PKCε and PKCδ work through, being an inhibitory action of PKCε on RhoA during proplatelets (ppt) formation in the mouse model while, in the human MK differentiation, platelets production is regulated by PKCδ through Bcl-xL. In this dissertation it is also demonstrated how in an aberrant megakaryocytopoiesis, as in the pathologic model of primary myeloproliferative neoplasm (PMF), PKCε is strongly deregulated and it results in an altered Bcl-xL expression. A forced down-modulation of this kinase restores a normal MK differentiation and ppt maturation. Therefore, the data presented show that PKCε and PKCδ play a key role in proper megakaryocyte maturation and that PKCε could be a potential new therapeutic target for PMF.

L'ingegneria tissutale è un campo multidisciplinare in rapida crescita che si avvale delle scienze fisiche, ingegneristiche e della vita per sviluppare ed ottenere cellule funzionali, tessuti ed organi per riparare, sostituire o migliorare le funzioni biologiche perse a causa di anomalie congenite, traumi, malattie o invecchiamento. Nell’ambito della ricostruzione della cartilagine articolare sono stati fatti notevoli passi in avanti ma la soluzione per il ripristino completo del tessuto sembra ancora essere lontana. Nella prima parte di questo lavoro è stata valutata la capacità di uno scaffold a base di gelatina di indirizzare le ADSCs verso un differenziamento in senso condrogenico. Successivamente, con lo scopo di migliorare il grado differenziamento e diminuire i costi associati all’utilizzo di fattori differenziativi, l’attenzione è stata posta sullo sviluppo di un biomateriale a base di alginato e ioni cobalto in modo da mimare e sfruttare le caratteristiche fisiche della cartilagine piuttosto che il suo intorno chimico. In ultimo, è stato sviluppato un sistema low cost per la produzione di chip microfluidici sfruttabili per la realizzazione di sistemi micrometrici per incapsulazione cellulare. / Tissue engineering is an interdisciplinary and multidisciplinary field that aims at the developmentof biological substitutes that restore, mantain, or improve tissue function. Concerning the articular cartilage many improvments were made, but the complete tissue restoration approach still lacking. In the first part of this work, it was evaluated the ability of a gelatin scaffold to promote the condrogenic differentiation of ADSCs. Successively, in order to obtain a low cost sistem, a based alginate/Cobalt scaffold was designed with the aim to take advantage of the physical features of the cartilage tissue. Finally, it was developted a cost effective method to produce microfluidic chips with the aim to obtain micro-systems for cell encapsulation.

Peripheral T-cell lymphomas-not otherwise specified (PTCL/NOS) are the most common T-cell neoplasms. This study sought to reshape the PTCL/NOS sub-classification (including its two main morphological variants, Lennert lymphoma, LL, and Follicular variant, F-PTCL) based on the correspondence between their molecular features and those of different functional T-cell subsets, also assessing the clinical impact of such an approach. We found that PTCLs/NOS could be divided into groups corresponding to T-cell subsets differently reliant on transcription regulators including mTOR and FOXP3, and identified minimal gene sets discriminating among these groups. Notably, by grouping tumors according to their dependency on master regulators of T-lymphocyte fate, we identified three groups (T-cytotoxic, Treg/TFH, and other-T-helper) characterized by specific genetic patterns and significantly different clinical outcomes. Immunohistochemistry partially substituted for the molecular analysis by consistently recognizing only Treg and TFH cases. Finally, targeted inhibition of MTOR in T-helper cases (that were characterized by genetic lesions targeting the pathway) was proved to be effective ex vivo. We conclude that PTCL/NOS can be divided into subgroups corresponding to different cellular counterparts, characterized by different genetic patterns and possibly sensitivity to specific therapeutic approaches. Furthermore, we identified different gene and microRNA signatures for LL capable of differentiating it from other PTCL/NOS and enriched in cytotoxic function. Moreover, PI3K/Akt/mTOR pathway emerged as novel therapeutic targets for LL. Additionally, LL showed some differences with other PTCL/NOS in terms of clinical features, all supporting its recognition as a distinct entity. Besides, we found that F-PTCL has a distinct molecular signature more similar to PTCL/NOS rather than AITL, and therefore cannot be included among AITLs at least based on GEP, although this necessities more genetic studies. Overall, these results may impact on PTCL classification as well as on future studies aimed to define the more appropriate therapeutic strategy for each identified subgroup/entity.

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Made available in DSpace on 2014-06-11T19:30:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-03-30Bitstream added on 2014-06-13T20:40:59Z : No. of bitstreams: 1 rosa_mm_dr_rcla.pdf: 3170631 bytes, checksum: ed110aae1aedd405aafb7422bf045267 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / No presente trabalho foi estudada a anatomia floral de Actinocephalus polyanthus, Leiothrix fluitans, Paepalanthus chlorocephalus, P. flaccidus e Rondonanthus roraimae, pertencentes à Eriocaulaceae, e de Cephalostemon riedelianus, pertencente à Rapateaceae, procurando contribuir com o maior conhecimento de representantes de Poales. Nas Eriocaulaceae, as flores estaminadas e pistiladas são trímeras na maioria das espécies e dímeras em P. flaccidus. Nas flores estaminadas de todas as espécies, o androceu é isostêmone, e em A. polyanthus, L. fluitans, P. chlorocephalus e P. flaccidus são encontrados estaminódios escamiformes microscópicos adnatos à base dos filetes. Nas flores pistiladas, os estaminódios são reduzidos, adnatos à base das pétalas, e em R. roraimae são lineares, alongados e vascularizados. As flores pistiladas de todas as espécies de Eriocaulaceae apresentam estiletes sólidos, desprovidos de vascularização e intercalados por apêndices nectaríferos que são vascularizados pelos feixes dorsais dos carpelos. Os apêndices nectaríferos são homólogos aos estiletes. A presença de androceu isostêmone pode ser considerada como um caráter derivado em Eriocaulaceae a partir da redução das peças florais de um provável ancestral com androceu diplostêmone. A ausência de estaminódios nas flores estaminadas de R. roraimae e a presença de estaminódios vascularizados nas suas flores pistiladas, indicam sua provável posição basal em relação aos demais representantes da subfamília Paepalanthoideae. A presença de apêndices nectaríferos pode ser considerada tanto um caráter derivado como um caráter ancestral em Eriocaulaceae a partir da modificação do gineceu do provável ancestral. Em relação à Rapateaceae, as flores de C. riedelianus são perfeitas, trímeras e apresentam androceu diplostêmone... / In order to contribute to a better knowledge of Poales specimens, the present work studied the floral anatomy of Actinocephalus polyanthus, Leiothrix fluitans, Paepalanthus chlorocephalus, P. flaccidus and Rondonanthus roraimae, belonging to the Eriocaulaceae, and of Cephalostemon riedelianus, belonging to the Rapateaceae. All the staminate and pistillate flowers of these Eriocaulaceae are trimerous except those of P. flaccidus, which are dimerous. All the staminate flowers of these species have an isostemonous androecium and A. polyanthus, L. fluitans, P. chlorocephalus and P. flaccidus present microscopic, scalelike staminodes adnate to the filament base. The staminodes of the pistillate flowers are reduced and adnate to the petal basis, except those of R. roraimae that are linear, elongated and vascularized. The pistillate flowers of all these Eriocaulaceae species present solid styles, devoid of vascularization and interspersed with nectariferous appendages, which are vascularized by the dorsal bundles of the carpels. The nectariferous appendages are homologous to the styles. The presence of an isostemonous androecium can be considered in Eriocaulaceae as a derived feature from the reduction of the floral parts from a probable ancestor with a diplostemonous androecium. The absence of staminodes in the staminate flowers of R. roraimae and the presence of vascularized staminodes in its pistillate flowers indicate its probable basal position in relation to other specimens of the subfamily Paepalanthoideae. In Eriocaulaceae, the presence of nectariferous appendages could be considered either as an ancestral feature or as a derived one from the modification of the gynoecium of a probable ancestor. With regard to the Rapateaceae, the flowers of C. riedelianus are perfect, trimerous and present a diplostemonous androecium... (Complete abstract, click electronic address below)

Made available in DSpace on 2014-06-11T19:30:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-12-18Bitstream added on 2014-06-13T19:19:47Z : No. of bitstreams: 1 bieras_ac_dr_rcla.pdf: 7881450 bytes, checksum: 1c1c258fdd1298eeff655a6a178f96c8 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Com o objetivo reconhecer os padrões morfológico e anatômico predominantes para as folhas de dicotiledôneas do cerrado, foram estudadas a morfologia de 70 espécies e a anatomia de 30 espécies arbóreo-arbustivas representativas da flora desse bioma no estado de São Paulo. As folhas são na sua maioria simples, variando de mesófila a microfila; apresentam lâminas elípticas, com margens inteiras e com bases e ápices convexos. Quanto à organização anatômica, em geral, as folhas de cerrado são dorsiventrais, hipostomáticas, encontram-se recobertas por tricomas e por espessas camadas de cera e de cutícula e possuem tecido vascular circundado por fibras. O tecido paliçádico é desenvolvido, sendo comum no mesofilo a ocorrência de células esclerificadas, distribuídas esparsamente, e de idioblatos contendo fenóis e cristais. Os resultados anatômicos encontrados foram comparados com descrições da literatura para outros representantes desse bioma e das famílias a que essas espécies pertencem. O possível significado desses caracteres é discutido dentro de um contexto adaptativo e evolutivo para o bioma cerrado. / With the objective of recognizing the commonest leaf pattern of the trees and shrubs of the cerrado vegetation, we studied the leaf morphology of 70 dicotyledons and the leaf anatomy of 30 species well represented in this bioma. The leaves are mostly simple, vary from mesophyll to microphyll, and the blades are eliptical with entire edges and with convex base and apex. They are mostly dorsiventral and hypostomatic, and covered by trichomes and thick layer of wax and cuticle; the vascular bundles are surrounded by a sheath of fibers. The mesophyll has a developed palisade tissue, dispersed sclerified cells and idioblasts bearing crystals and phenolic compounds. The anatomical results were compared with the described for other species from the same biome and for the families that the studied species belong. The possible meaning of these features is discussed under an adaptative and evolutive point of view.

coordenadoria de aperfeiÃoamento de pessoal de ensino superior / O cÃncer gÃstrico à a quarta neoplasia mais comum em todo o mundo e a segunda causa de mortalidade por cÃncer. Apesar do tratamento com cirurgia e quimioterapia, a sobrevida global em cinco anos de pacientes com cÃncer gÃstrico permanece baixa, uma possÃvel explicaÃÃo para menor eficÃcia da terapia à a presenÃa de cÃlulas tronco cancerosas. VÃrios marcadores, incluindo LGR5 e LGR6, tÃm sido relatados como marcadores de cÃlulas- tronco, normais e cancerosas. LGR5 e LGR6 sÃo proteÃnas transmembranas, do grupo da proteÃna G, ricas em resÃduos leucina, que participam nas vias de sinalizaÃÃo das cÃlulas e ativam fatores de transcriÃÃo relacionados com a formaÃÃo de tumores. O objetivo deste trabalho foi avaliar a imunoexpressÃo de LGR5 e LGR6 como possÃveis marcadores de cÃlulas-tronco no cÃncer gÃstrico primÃrio, metÃstases linfonodais e mucosa gÃstrica histologicamente normal, atravÃs das tÃcnicas de microarranjo tissular (tissue microarray) e imuno-histoquÃmica. O estudo, de carÃter transversal e observacional realizou-se a partir de oitenta e oito (88) peÃas de gastrectomias devido a carcinomas gÃstricos, realizadas no Hospital UniversitÃrio Walter CantÃdio, que fazem parte dos Arquivos do ServiÃo de Patologia e Medicina Legal da Universidade Federal do CearÃ. As relaÃÃes entre a expressÃo diferencial de LGR5 e LGR6 e caracterÃsticas clinico-patolÃgicas foram avaliadas pelo teste do qui-quadrado ou teste exato de Fisher. Um valor de p < 0,05 foi considerado estatisticamente significativo. Foramconsiderados positivos (a partir de relatos prÃvios) casos que apresentaram uma ou mais cÃlulas com imunomarcaÃÃo citoplasmÃtica ou membranar. LGR5 mostrou-se, neste estudo, ser um potencial biomarcador, mais caracterÃstico de cÃlulas-tronco tumorais e nÃo tumorais, do que LGR6. Ambos sÃo expressos em tecido tumoral e nÃo tumoral, com predomÃnio em mucosa histologicamente normal, em terÃo basal da espessura epitelial. LGR5 positivo predominou no histotipo difuso e LGR6 no tipo intestinal de carcinomas gÃstricos. Nos casos positivos dos dois biomarcadores, cÃlulas marcadas sÃo pouco frequentes ou raras, no total de cÃlulas da massa tumoral e da mucosa gÃstrica normal. LGR6 esteve presente no tumor gÃstrico primÃrio em nÃmero de cÃlulas muito superior ao encontrado nas respectivas metÃstases linfonodais, diferenÃa nÃo observada em relaÃÃo a LGR5. Com exceÃÃo da relaÃÃo de cada biomarcador com um histotipo especÃfico, jà citada, nÃo houve relaÃÃo entre as demais variÃveis clinico-patolÃgicas e a expressÃo de LGR5 e LGR6. / Gastric cancer is the fourth most common neoplasm in the world and the second leading cause of cancer mortality. Despite the treatment with surgery and chemotherapy, the overall five-year survival of patients with gastric cancer remains low, a possible explanation for less effective therapy is the presence of cancer stem cells. Several markers, including LGR5 and LGR6, have been reported as markers of normal and cancerous stem cells. LGR5 and LGR6 are transmembrane proteins of the G protein group, rich in leucine residues, that participate in cell signaling pathways and activate transcription factors related to tumor formation. The objective of this work was to evaluate the immunoexpression of LGR5 and LGR6 as possible markers of stem cells in primary gastric cancer, lymph node metastases and histologically normal gastric mucosa through tissue microarray and immunohistochemistry techniques. The cross-sectional and observational study was carried out from eighty-eight (88) pieces of gastrectomies due to gastric carcinomas performed at the Walter CantÃdio University Hospital, which are part of the Archives of the Service of Pathology and Legal Medicine of the Federal University Of CearÃ. The relationships between LGR5 and LGR6 differential expression and clinical-pathological characteristics were assessed by the chi-square test or Fisher's exact test. A value of p <0.05 was considered statistically significant. Positive (from previous reports) were considered cases that presented one or more cells with cytoplasmic or membrane immunostaining. LGR5 was shown to be a potential biomarker, more characteristic of tumor and non-tumor stem cells, than LGR6 in this study. Both are expressed in tumoral and non-tumoral tissue, predominantly in histologically normal mucosa, in a basal third of epithelial thickness. Positive LGR5 predominated in the diffuse histotype and LGR6 in the intestinal type of gastric carcinomas. In the positive cases of the two biomarkers, marked cells are infrequent or rare, in the total cells of the tumor mass and the normal gastric mucosa.

The present study evaluated the effects of L lysine in intestinal and urothelial epithelia in augmentation of bladder and ureterosigmoidostomy in rats.A total of 66 female rats 9 weeks old were split divided in to 9 experimental groups. The animals from groups: I , II and III were subjected to bladder augmentation with colon segment (AV) and treated with L lysine, celecoxib and H2O, respectively. The groups: IV, V and VI were subjected to ureterosigmoidostomy (US) treated with L lysine, celecoxib and H2O in that order. Groups: VII, VIII and IX (non- operated controls) received L lysine, celecoxib and H2O respectively. The dosage of L lysine was 150mg/ kg/ body weight and the celecoxib was 20mg/kg/ body weight. The effects of L lysine on the colon epithelium of rats subjected to US was initially evaluated by analysis Aberrant Cripts Foci (ACF) at stereostopic microscopy, after fixation with formaldehyde and staining with methylene blue. Followed by histopathological study of ureteral epithelium, colonic and bladder in all groups, stained with hematoxylin and eosin and PAS Alcian Blue. Rare ACF were found in all mice with US and compared between groups. There was no statistically significant difference between groupsOn histopathology with hematoxylin and eosin, mild to moderate hyperplasia was observed in intestinal and urothelial epithelia at the site of anastomosis in all animals submitted to cystoplasty (Groups I and III), but transitional metaplasia of the intestinal glandular epithelium was more accentuated in Group I (p=0.045). There were no inflammatory cells, dysplasia or atypia. In epithelia of the left ureter and colon of mice with US a mild inflammatory infiltrate composed of lymphocytes, moderate to severe intensity, had urothelial hyperplasia of moderate to severe intensity in all the ureters, three polyps in different ureters and inflammatory polyps in colon . There were no dysplasia or atypia. The staining with Alcian Blue, noticed an important decrease of goblet cells and mucin in colon in all operated rats. In the histopathology there is similarity in the quality and quantity of injuries. Thus, we conclude that L-Lysine did not influence the carcinogenesis of intestinal epithelia, and urothelial of rats submitted to US and AV with colon segment, at times, doses and methods evaluated.However, the L-lysine stressed the "urothelial metaplasia" in intestinal segment bladder and ureter transitional hyperplasia in rats subjected to ureterosigmoidostomies. / Avalia os efeitos da L lisina na carcinogÃnese de epitÃlios intestinal e urotelial nasampliaÃÃes vesicais e ureterossigmoidostomias em ratas. O total de 66 ratas com nove semanas de idade foi dividido em nove grupos. Os animais dos grupos I, II e III foram submetidas a ampliaÃÃo vesical com segmento de colo (AV) e tratados com L lisina, celecoxibe e H2O, respectivamente. Os do grupo IV, V e VI foram submetidos a ureterossigmoidostomia (US) e tratados com L lisina, celecoxibe e H2O nesta ordem. Os grupos VII, VIII e IX (controles nÃo operados), receberam L lisina, celecoxibe e H2O, respectivamente. A dose de L lisina foi de 150 mg/kg/peso e o celecoxibe foi de 20 mg/kg/peso. Os efeitos da L lisina no epitÃlio do colo de ratos submetidos a US foi inicialmente avaliado pela anÃlise de FCA (focos de criptas aberrantes) a microscopia estereoscÃpica apÃs fixaÃÃo com formol e coloraÃÃo pelo azul de metileno. Seguiu-se estudo histopatolÃgico dos epitÃlios ureterais, cÃlicos e vesicais em todos os grupos, corados pela hematoxilina e eosina e PAS Alcian Blue. Foram encontrados raros FCA, em todas as ratas com US e na comparaÃÃo entre os grupos. NÃo ocorreu diferenÃa estatisticamente significantes entre os grupos. No estudo histopatolÃgico, com hematoxilina e eosina de epitÃlios cÃlicos e vesicais de animais com AV, foram observados hiperplasia urotelial leve a moderada em todos os animais submetidos a cistoplastia, em regiÃo de anastomoses colovesical e maior nÃmero de animais apresentaram âmetaplasia transicionalâ em epitÃlio glandular intestinal no grupo I (p= 0,045). NÃo havia cÃlulas inflamatÃrias, displasias ou atipias. Nos epitÃlios de ureter esquerdo e colo das ratas com US, em meio a infiltrado inflamatÃrio constituÃdo por linfÃcitos, de moderada a acentuada intensidade, havia hiperplasia urotelial de moderada a acentuada, trÃs polipos em distintos ureteres e um polipo inflamatÃrio em colo. NÃo havia displasias ou atipias. Quanto à coloraÃÃo pelo Alcian Blue, notou-se importante diminuiÃÃo de cÃlulas caliciformes e de mucinas em colo, em todas as ratas operadas. Desta forma, conclui-se que a L lisina nÃo influenciou na carcinogÃnese do epitÃlio intestinal das ureterossigmoidostomias em ratas nos tempos e doses, bem como pelo mÃtodo de avaliaÃÃo de criptas aberrantes. A L lisina acentuou a âmetaplasia urotelialâ em segmento intestinal de ampliaÃÃes vesicais e a hiperplasia transicional no ureter das ratas submetidas a ureterossigmoidostomias.