No description available.

No description available.

The heterogeneity of salivary gland neoplasms have made classification and prognostication of these tumours sometimes difficult, and the in­troduction of techniques, such as enzyme and carbohydrate histochemis­try and electron microscopy have only to a certain extent increased our knowledge in these respects. In the present study immunohistochemical methods have been used to identify intermediate filament proteins (IFP) in normal fetal and adult parotid glands, as well as in salivary neo­plasms. The intermediate filaments (IF) make up the cytoskeleton in eucaryotic cells. Epithelial tissue contains IF composed of different cytokeratins (CK 1-19) whilst mesenchymal tissue generally contains IF composed of vimentin, and the IFP pattern is very stable even during cell transformation. It would thus be possible to further clarify the histogenesis of salivary neoplasms by identifying IFP, in addition the IFP pattern would probably be useful in tumour typing. Furthermore, ultrastructural cytochemical studies, microspectorphotometry on nuclear DNA as well as enzyme secretory studies of certain tumour types were carried out, in order to further characterize the biology of salivary neoplasms. The immunohistochemical investigations showed that in normal parotid tissue, the different cell types differed in IFP expression: acinar cells express mainly CK 18 and myoepithelial cells mainly CK 17 and 19, whilst duct cells contained a broad range of CK. Vimentin could in ad­dition to CK be detected in myoepithelial cells and basal cells of ex­cretory ducts. Fetal parotid cells showed a similar CK pattern as mature duct cells. In addition, vimentin could be found in some basal cells of the terminal tubules of the fetal glands. Salivary neoplasms could be divided into three types with regard to their IFP pattern:  Acinic cell carcinomas showed a CK-pattern similar to normal acinar cells but a co-expression of CK and vimentin was present in some cells.  Adenoid cystic carcinomas, mixed tumours and basal cell adenomas showed a CK-pattern of normal duct or myoepithelial cells. The peri­pheral cells were also vimentin positive. 3. Mucoepidermoid carcinomas and adenocarcinomas had a similar CK-pattern as duct cells, and no tu­mour cells contained vimentin. This indicates that typing of IFP may be useful for subgrouping of salivary neoplasms. By stereological measurements, the cells of acinic cell carcinomas were found to be very similar to normal parotid acinar cells. Furthermore, they contained amylase and after stimulation by norepiphrine a secre­tory response was induced, with a rise in intracellular cAMP as well as a release of amylase. By single cell measurements of nuclear DNA con­tent, no difference was found between acinic cell carcinomas with de­finite metastasis and those without recurrence, both in paraffin sec­tions and cytological smears. / digitalisering@umu.se

No description available.

No description available.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2018-09-24T15:13:44Z No. of bitstreams: 1 2017 - Enely Maris da Silveira Firmiano.pdf: 12307016 bytes, checksum: d58bee7e5ddc0113fa072ab4e9aabc29 (MD5) / Made available in DSpace on 2018-09-24T15:13:44Z (GMT). No. of bitstreams: 1 2017 - Enely Maris da Silveira Firmiano.pdf: 12307016 bytes, checksum: d58bee7e5ddc0113fa072ab4e9aabc29 (MD5) Previous issue date: 2017-08-29 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Cap. I The aim of the current study is to investigate the histological and histochemical description, as well as the presence and immunohistochemical distribution of cytoskeleton proteins such as alpha smooth muscle actin, desmin and vimentin in the ovaries, testes and epididymis of Enyalius bilineatus, Enyalius perditus and Urostrophus vautieri. These species are representatives of family Leiosauridae, in order to help developing a database on the reproductive biology of these lizards, which could be compared to that of other reptile species and vertebrates. The herein studied specimens are deposited in the Herpetological Collection of Federal University of Juiz de Fora. The animals? ovaries, epididymis and testes were removed and sent to the Histology and Embryology Laboratory (UFRRJ), where they were fixed, processed and then subjected to histological, histochemical and immunohistochemical techniques. The ovaries of these leiosauridae are functionally-active paired organs filled with ovarian follicles at different follicular development stages. These organs are covered by a simple cubic epithelium, which presents an underlying connective tissue layer called tunica albuginea. The testes are covered by the tunica albuginea and hold septa forming lobes where the seminiferous tubules are located in. The epididymis of these lizards are convolute ducts internally coated by an epithelium, whose shape ranges from simple cylindrical to simple cubic with stereocilia. The immunohistochemical analysis applied to the ovaries of the herein studied lizards showed moderate to strong reaction to the alpha smooth muscle actin in the teak layer and in endothelial cells of blood vessels. With respect to the species? testes, this cytoskeleton protein showed strong reaction in the tunica albuginea, interstitial tissue, and in endothelial cells of blood vessels. As for the epididymis, the alpha-actin showed immunohistochemical reaction in the capsule and in the interstitial tissue of the lizards. The antibody ?desmin? showed strong immunostaining in the albuginea and ovarian stroma of E. perditus, only; however, the endothelial cells in the blood vessels of the three herein analyzed species showed positive reaction to this intermediate filament. Desmin showed immunoreactivity in the testicular albuginea and in the endothelial cells of the blood vessels in the herein analyzed leiosauridae testes; however, only E. bilineatus showed immunostaining in the interstitial region. Desmin showed positive immunoreaction in the epididymis of E. perditus and E. bilineatus, only. Vimentin did not show immunostaining in the ovaries, testes and epididymis of the three analyzed lizards. The consistent results found in the current study provide additional data on the reproductive biology of the investigated species. These data may be used for phylogenetic and behavioral comparisons between the herein studied species and other species belonging to family Leiosauridae or other reptiles Cap. II The aim of the current study is to investigate the histological description, as well as the presence and immunohistochemical distribution of alpha smooth muscle actin, desmin and vimentin in the testes and epididymis of Enyalius bilineatus, Enyalius perditus and Urostrophus vautieri, which are representatives of family Leiosauridae, in order to help developing a database on the reproductive biology of these lizards, which could be compared to that of other reptile species and vertebrates. The herein studied specimens are deposited in the Herpetological Collection of Federal University of Juiz de Fora. The animals? epididymis and testes were removed and sent to the Histology and Embryology Laboratory (UFRRJ), where they were fixed, processed and then subjected to histological and immunohistochemical techniques. The shape of the testes varies reasonably among the studied leiosauridae. These organs are covered by the tunica albuginea and hold septa forming lobes, where the seminiferous tubules are located in. The epididymis of these lizards are convolute ducts internally coated by an epithelium, whose shape ranges from simple cylindrical to simple cubic with stereocilia. The immunohistochemical analysis applied to the testes of the herein studied lizards showed strong positive reaction to the alpha smooth muscle actin in the tunica albuginea, interstitial tissue, and in endothelial cells of blood vessels. Desmin showed strong immunoreactivity in the testicular albuginea of E. bilineatus and moderate immunoreactivity in E. perditus and U. vautieri; however, only E. bilineatus showed immunostaining in the interstitial tissue region. The endothelial cells of blood vessels showed positive immunoreaction to desmin in the three herein analyzed species. The epididymis showed moderate immunohistochemical reaction to alpha smooth muscle actin in the capsule and in the interstitial tissue, whereas desmin showed positive immunoreaction in the epididymis of E. perditus and E. bilineatus, only. Vimentin did not show immunostaining in the testes and epididymis of the three analyzed lizards. The consistent results found in the current study provide additional data on the reproductive biology of the investigated species. These data may be used for phylogenetic and behavioral comparisons between the herein studied species and other species belonging to family Leiosauridae or other reptiles. / Cap. I Este trabalho investiga, al?m da descri??o histol?gica e histoqu?mica, a presen?a e distribui??o imuno-histoqu?mica de alfa-actina de m?sculo liso, desmina e vimentina nos ov?rios, test?culos e epid?dimos de Enyalius bilineatus, Enyalius perditus e Urostrophus vautieri, representantes da fam?lia Leiosauridae, para ajudar a formar uma base de dados sobre a biologia reprodutiva destes lagartos que possam ser comparados com outras esp?cies de r?pteis, bem como com outros vertebrados. Os esp?cimes estudados encontram-se depositados na Cole??o Herpetol?gica da UFJF. Os ov?rios, epid?dimos e test?culos dos animais foram removidos e encaminhados ao Laborat?rio de Histologia e Embriologia (UFRRJ), onde foram fixados, processados e ent?o submetidos ?s t?cnicas histol?gicas, histoqu?micas e imuno-histoqu?micas. Os ov?rios destes leiosaur?deos s?o ?rg?os pareados, repletos de fol?culos ovarianos, em distintos est?gios de desenvolvimento folicular. Estes s?o revestidos por epit?lio c?bico simples, com uma camada de tecido conjuntivo subjacente, a t?nica albug?nea. Os test?culos encontram-se envolvidos pela t?nica albug?nea e em seu interior cont?m septos que formam l?bulos, nos quais se localizam os t?bulos semin?feros. Os epid?dimos destes lagartos s?o ductos convolutos, revestidos internamente por um epit?lio que varia de cil?ndrico simples a c?bico simples com estereoc?lios. A imuno-histoqu?mica, nos ov?rios dos lagartos estudados, mostra uma rea??o moderada a acentuada para alfa-actina de m?sculo liso na camada da teca e em c?lulas endoteliais dos vasos sangu?neos, enquanto nos test?culos destas esp?cies nota-se uma acentuada rea??o para esta prote?na do citoesqueleto, na t?nica albug?nea, no tecido intersticial e nas c?lulas endoteliais dos vasos sangu?neos. Nos epid?dimos observa-se imunomarca??o para alfa-actina na c?psula e no tecido intersticial destes lagartos. Quanto ao anticorpo desmina, apenas em E. perditus, foi poss?vel imunorrea??o na albug?nea e no estroma ovariano, no entanto, nas tr?s esp?cies analisadas, as c?lulas endoteliais dos vasos sangu?neos demonstraram rea??o positiva a este filamento intermedi?rio. Nos test?culos dos leiosaur?deos estudados, a desmina foi imunolocalizada na albug?nea testicular e nas c?lulas endoteliais dos vasos sangu?neos, entretanto, apenas em E. bilineatus ocorreu imunomarca??o, na regi?o do interst?cio. Nos epid?dimos, a desmina demonstrou imunorrea??o positiva apenas em E. perditus e E. bilineatus. N?o houve rea??o imuno-histoqu?mica para vimentina nos ov?rios, test?culos e epid?dimos dos tr?s lagartos analisados. Este estudo obteve resultados consistentes, fornecendo assim mais dados sobre a biologia reprodutiva das esp?cies aqui investigadas, que poder?o ser utilizados para compara??es filogen?ticas e comportamentais com outras esp?cies da fam?lia Leiosauridae e outros r?pteis. Cap. II Este trabalho investiga, al?m da descri??o histol?gica, a presen?a e distribui??o imuno-histoqu?mica de alfa-actina de m?sculo liso, desmina e vimentina nos test?culos e epid?dimos de Enyalius bilineatus, Enyalius perditus e Urostrophus vautieri, representantes da fam?lia Leiosauridae, para ajudar a formar uma base de dados sobre a biologia reprodutiva destes lagartos que possam ser comparados com outras esp?cies de r?pteis, bem como com outros vertebrados. Os esp?cimes estudados encontram-se depositados na Cole??o Herpetol?gica da Universidade Federal de Juiz de Fora. Os epid?dimos e test?culos dos animais foram removidos e encaminhados ao Laborat?rio de Histologia e Embriologia (UFRRJ), onde foram fixados, processados e ent?o submetidos ?s t?cnicas histol?gicas e imuno-histoqu?micas. O formato dos test?culos varia razoavelmente entre os leiosaur?deos estudados. Estes ?rg?os encontram-se envoltos pela t?nica albug?nea e no seu interior cont?m septos que formam l?bulos incompletos, nos quais se localizam os t?bulos semin?feros. Os epid?dimos destes lagartos s?o ductos convolutos. Estes ductos s?o revestidos internamente por um epit?lio que varia de cil?ndrico simples a c?bico simples com estereoc?lios. A imuno-histoqu?mica revela uma acentuada rea??o positiva para alfa-actina de m?sculo liso nos test?culos dos leiosaur?deos analisados, na t?nica albug?nea e no tecido intersticial, assim como, nas c?lulas endoteliais dos vasos sangu?neos. A desmina apresentou acentuada imunorreatividade na albug?nea testicular em E. bilineatus e moderada em E. perditus e U. vautieri, entretanto, na regi?o do tecido intersticial, apenas em E. bilineatus ocorreu imunomarca??o. Nas tr?s esp?cies analisadas, as c?lulas endoteliais dos vasos sangu?neos demostraram imunorrea??o positiva para desmina. Nos epid?dimos observou-se rea??o imuno-histoqu?mica moderada para alfa-actina de m?sculo liso na c?psula e no tecido intersticial, enquanto a desmina demonstrou imunorrea??o positiva apenas em E. perditus e E. bilineatus. N?o houve imunomarca??o para vimentina nos test?culos e epid?dimos dos tr?s lagartos analisados. Este estudo obteve resultados consistentes, fornecendo assim mais dados sobre a biologia reprodutiva das esp?cies aqui investigadas, que poder?o ser utilizados para compara??es filogen?ticas e comportamentais com outras esp?cies da fam?lia Leiosauridae e outros r?pteis.

Indiana University-Purdue University Indianapolis (IUPUI) / Giant axonal neuropathy (GAN) is a rare genetic disease that causes progressive damage to the nervous system. Neurons in GAN patients develop an abnormal organization of cytoskeletal proteins called intermediate filaments (IFs), which normally provide strength and support for the overall cell structure. The irregular IF structure in GAN patient neurons leads to a progressive loss of motor skills in children and subsequent death in adolescence. GAN is caused by reduced levels of the gigaxonin (Giga) protein. Giga functions to control the degradation of other cellular proteins, and the loss of Giga in GAN cells results in significantly elevated levels of the galectin-1 (Gal-1) protein. Gal-1 stabilizes the active form of the Ras signaling protein, which functions as a molecular switch to regulate the phosphorylation and subsequent organization of IFs. The connection between these pathways led us to propose that Giga regulates IF phosphorylation and structure by modulating Ras signaling through the degradation of Gal-1. Using GAN patient cells, we demonstrated that restoring Giga reduced Gal-1 protein levels, decreased IF phosphorylation, and reestablished normal IF organization. Similar effects of reduced IF phosphorylation and improved IF structure were also obtained in GAN cells by directly decreasing the protein levels of either Gal-1, or downstream Ras signaling proteins. Taken together, these results demonstrate that the loss of Giga induces Gal-1 mediated activation of Ras signaling, thereby leading to the increased IF phosphorylation and abnormal IF structure observed in GAN cells. Identification of aberrant Ras signaling is significant because it is the first to specify a mechanism by which the loss of Giga leads to the development of GAN and provides targets for novel drug therapies for the treatment of this currently immedicable genetic disease.

Orientador: Antonio Condino Neto / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-13T22:05:12Z (GMT). No. of bitstreams: 1 Talero_LuisAlbertoPedroza_D.pdf: 1538530 bytes, checksum: 6dc94ec71cdd6f03d096be015a2b1757 (MD5) Previous issue date: 2009 / Resumo: A Poliendocrinopatia autoimune associada a candidíase e distrofia ectodérmica (APECED) é um síndrome caracterizado pela presença de pelo menos dois sintomas clínicos, endocrinopatia autoimune, sendo que as mais comuns são hipoparatiroidismo, doença de Addison, além de candidíase mucocutânea crônica. É também comum nos pacientes o desenvolvimento de distrofia ectodérmica, como distrofia nas unhas ou alopécia. O APECED é produzido por mutações no gene AIRE, que codifica uma proteína com propriedades reguladoras na transcrição de proteínas ectópicas no timo, o que estaria envolvido na seleção negativa de células T auto-reativas, e conseqüentemente no desenvolvimento da doença autoimune. No entanto a associação da deficiência da proteína AIRE com a suscetibilidade a candidíase ou a distrofia ectodérmica permanecem obscuras. No presente trabalho, investigamos a possibilidade que esta associação esteja envolvida com a expressão e função da proteína AIRE no ambiente extra-tímico. Usando células de sangue periférico de pacientes com mutações no AIRE, e a técnica de SiRNA para silenciar este gene em células de linhagem mielomonocítica THP-1, demonstramos que a proteína AIRE é importante para a resposta via KF-kB dos TLRs e Dectina-1, sendo que AIRE está presente num complexo com Dectina-1, Syk e Card-9, formado após o estímulo com Curdlan. Além disso demonstramos que a formação deste complexo pode acontecer no citoplasma ou membrana citoplasmática, uma vez que após este estímulo, a proteína AIRE é exportada do núcleo permanecendo temporariamente na membrana. Finalmente usando a técnica de espectroscopia de massa e microscopia confocal, mostramos que AIRE interage com a proteína Queratina 17, tanto em células THP-1 como em células HaCaT (linhagem de queratinócitos), quando as células entram num estágio de espraiamento e migração. Assim, a presença da proteína AIRE na via de sinalização da Dectina-1, pode estar relacionada com a susceptibilidade a infecções crônicas por C. albicans observada nestes pacientes. A resposta imune via Dectina-1 é importante na resposta a este fungo e defeitos na molécula CARD9 e Dectina-1 podem estar associados a Candidíase mucocutânea crônica. Por outro lado, a descrição da associação de AIRE com K17 pode ser relevante, já que pacientes com mutações no gene que codifica para a proteína K17 desenvolvem uma doença chamada paquioníquia congênita, caracterizada por distrofia das unhas e alopécia, características clínicas observadas também nos pacientes com APECED. Deste modo, neste trabalho apresentamos evidências que apontam para um novo papel funcional da proteína AIRE no ambiente extratímico, que poderia explicar em parte algumas características clínicas dos pacientes com APECED, como a elevada suscetibilidade a infecções por C. albicans, e a distrofia ectodérmica / Abstract: The autoimmune polyendocrinopathy candidiasis and ectodermal dystrophy (APECED) is characterized by the presence of two from three major clinical symptoms: Addison's disease, and/or hypoparathyroidism, and/or chronic mucocutaneous candidiasis. These patients develop also ectodermal dystrophies like nail dystrophy and alopecia. APECED is caused by mutations in the autoimmune regulator gene (AIRE). This gene encodes a protein with DNA binding capacity that can transcriptionally modulate ectopic peripheral tissue antigen (PTA) expression in the thymus, facilitating T cell negative selection. Defects in AIRE may be related with the development of multipleendocrine failure of autoimmune origin in patients with APECED. In spite of this, the role of AIRE deficiency in the C. albicans susceptibility or ectodermal dystrophy, common features in APECED patients, remains to be elucidated. In the present work we explored the hypothesis that candidiasis and ectodermal dystrophy are associated with the extra-thymic role of AIRE. For this we used peripheral blood mononuclear cells from APECED patients, and also THP-1 cells treated with SiRNA for AIRE gene to obtain AIRE deficient cells. We demonstrated that AIRE is required for Dectin-1- and TLR-ligand-induced inflammatory response and complexes with Dectin-1, Syk, and CARD9 after Curdlan stimulation. In addition, we showed that this complex formation takes place outside the nucleus, once that after Curdlan stimulation AIRE seems to be exported to the cytoplasm and transiently locate at the cytoplasmic membrane. Finally using mass spectra and confocal microscopy, we showed an interaction between AIRE and the intermediate filament protein Keratin-17, in both THP-1 cells and the keratinocyte cell line HaCaT. Therefore, the presence of AIRE protein in Dectin-1 pathway seems to be important on the C. albicans response, and the absence of this protein could be a risk factor important for developing candidiasis, commonly observed in APECED patients. This observation is supported by the fact that Dectin-1 is important for C albicans response, and also the recently description of mutations in Dectin-1 and CARD9 and its association with chronic mucocutaneous candidiasis. On the other hand, the description of AIRE and K17 association is important, since patients with defects on K17 gene develop congenital pachyonychia, a disease characterized by nail dystrophy and alopecia, also observed in APECED patients. Thus we provided evidence for a new role of AIRE protein in the extrathymic environment, which in may explain, at least in part, some of the common clinical features other than autoimmunity, observed in APECED patients / Doutorado / Saude da Criança e do Adolescente / Doutor em Saude da Criança e do Adolescente