Orientadora: Maria Angélica Miglino / Banca: Wilter Ricardo Russiano Vicente / Banca: Tatiana Carlesso dos Santos / Banca: Isaura Maria Mesquita Prado / Banca: Gilson Hélio Toniollo / Resumo: Este estudo utilizou 15 fêmeas bufalinas, provenientes de abatedouros frigoríficos dos estados do Maranhão, São Paulo e Paraná para estudo morfológico, macroscópico, microscópico e morfométrico das papilas mamárias. O material para estudo macroscópico foi pesado, medido e fotografado, e os resultados do peso papilar foram de: cranial direita (PCD) 20, 95g; caudal direita (PCaD) 21,49g; cranial esquerda (PCE) 23,52g e para caudal esquerda (PCaE) 21,35g. Os valores médios para comprimento do ducto papilar, número de pregas e comprimento da cisterna papilar foram de: 0,67cm; 5,5 e 2,17cm para PCD; 0,62cm; 6,0 e 2,26cm para PCaD; 0,71cm; 6,0 e 2,26cm para PCE e 0,74cm; 5,7 e 2,57cm para PCaE, respectivamente. A análise de correlação de Pearson indicou não existir relação entre o peso corpóreo e o peso glandular. Fragmentos das regiões proximal, média e distal foram colhidos e fixados em solução aquosa de formol 10%, cortados com 10, 20 e 40mm e corados em Azan, Picrocírus F3BA e Weigert-Van Gieson, para a análise mesoscópica. Cortes com 5mm corados em HE, tricrômico de Masson foram utilizados para a análise histológica e morfométrica. A estrutura histológica da papila mamária em búfalas mostrou-se similar aquela dos bovinos, a camada muscular é formada por fibras em duas disposições, a interna circular e a externa longitudinal, e com epitélio cúbico biestratificado em quase toda a sua extensão. Na porção média da papila a camada mais desenvolvida é a muscular com 22,90mm de espessura, a qual possui grande quantidade de tecido conjuntivo denso (7,10 mm). / Abstract: Fifteen Murrah female buffaloes, proceeding from abattoirs of Maranhão, São Paulo and Paraná states, were used in this study and submitted for morphologic, macroscopic, microscopic and morphometric analysis of the mammary papilla. The material for macroscopic study was weighed, measured and photographed, the results for papillary weight had been: cranial right (PCD) 20.95g; caudal right (PCaD) 21.49g; left cranial (PCE) 23.52g and for left caudal (PCaE) 21.35g. The average values for the papillary duct length, number of folds and length of the papillary cistern had been respectively for PCD: 0.67cm, 5.5, 2.17cm; PCaD: 0.62cm, 6.0, 2.26cm; PCE: 0.71cm, 6.0, 2.26cm and PcaE: 0.74cm, 5.7, 2.57cm. The analysis of de Pearson correlation coefficient test between corporeal weight and the glandular weight indicated not relation existence. Proximal, meddle and distal tissue samples had been collected and fixed in 10% formol watery solution and cut with 10-20-40mm and stained in Azan, Picrosirus F3BA and Weigert-Van Gieson, for the mesoscopic analysis. Cuts of 5mm and had stained in HE and Masson's trichrome were submitted to the morphologic and morphometric analysis. Buffalo's' mammary papilla structure revealed to be similar to the described for bovines, the muscular layer was composed by two bundles of smooth muscle fibers, the external circular layer and the longitudinal external one. The mucosa was bi-stratified cuboidal ephitelia in almost all its extension. The papilla meddle portion muscular layer was 22.90mm of thickness which contained large amount of connective tissue (7.10mm). / Doutor

A ausência ou perda da papila interdental cria deficiência estética, problemas fonéticos, impactação alimentar e gera muita expectativa ao paciente. Até o momento, o tratamento da ausência ou perda da papila interdental tem sido mal sucedido e não há estudos que indiquem que a regeneração da papila é um resultado previsível. O objetivo deste estudo foi avaliar a efetividade da injeção de gel de ácido hialurônico de origem não animal na redução ou eliminação da deficiência de papila entre dentes naturais comparativamente ao tratamento por meio de enxerto de tecido conjuntivo subepitelial. Foram avaliados neste estudo 20 sítios de 6 pacientes de ambos os sexos, com idade variável de 29 a 62 anos, apresentando deficiência de papila entre dentes naturais, na região anterior superior, em pelo menos dois dentes. Os 20 sítios tratados foram aleatoriamente divididos em dois grupos, de acordo com o tratamento para correção da deficiência de papila por meio de enxerto de tecido conjuntivo subepitelial (grupo controle) ou por meio de injeção de gel de ácido hialurônico (grupo teste). Um examinador único, calibrado, avaliou a distância da ponta da papila ao ponto de contato com auxílio de sonda periodontal milimetrada antes e aos 1, 3 e 6 meses após o tratamento. Além disso, foram investigados, nos sítios tratados, as medidas de profundidade de sondagem, nível de inserção, índice de sangramento do sulco, índice de placa, distância do ponto de contato à crista óssea alveolar, distância da ponta da papila à crista óssea alveolar e largura da papila. Os resultados demonstraram que aos 6 meses de pósoperatório o percentual de mudança na altura da papila foi maior no grupo teste (14,94% ± 21,35%) do que no grupo controle (-1,39% ± 31,46%), entretanto sem diferenças significantes entre os grupos (p> 0.05). Não houve variação estatisticamente significante na largura da papila antes e aos 4 meses após o tratamento nos grupos teste (p= 0.09) e controle (p= 0.16), assim como não houve variação significativa na distância entre a ponta da papila e a crista óssea alveolar. Houve melhora significativa do Índice de Estética Rosa (IER) observado aos 6 meses de acompanhamento em comparação com a condição inicial no grupo teste (p= 0.0078; Wilcoxon), enquanto que não houve mudança significativa no IER observado no grupo controle aos 6 meses de acompanhamento (p= 0.35). Os resultados obtidos permitiram concluir que o tratamento da deficiência de papila por meio de injeção de gel de ácido hialurônico promove melhora da deficiência de papila, similar aos resultados obtidos com o tratamento por meio de enxerto de tecido conjuntivo subepitelial, porém com melhora estética significativa relacionada especialmente às características de cor e textura do tecido relativamente aos tecidos moles adjacentes. / The absence or loss of interdental papilla creates an esthetic deficiency, phonetic problems and food impaction and generates a lot of expectation for the patient. Until now, the treatment for absence or loss of interdental papilla is unsuccessful e and there are no researches that show that the papilla regeneration is a predictable outcome. The aim of this study was to evaluate the effectivity of a non-animal originated hyaluronic acid injection in the reduction or elimination of papilla deficiency between natural teeth in comparison to a sub epithelial connective tissue graft treatment. The analysis was made on 20 sites in 6 patients, both genders, 29 - 62 years, showing deficiency in the papilla between natural teeth in the upper anterior region in at least two teeth. The 20 sites treated were randomly divided into two groups, according to the treatment by subepithelial connective tissue graft (control group) or by hyaluronic acid injection (test group). A single calibrated examiner evaluated the distance between the tip of the papilla to the contact point using a graduated periodontal probe before the treatment and 1, 3 and 6 months after it. Besides, it were investigated probing pocket depth, clinical attachment level, gingival bleeding index, plaque index, distance from papilla to alveolar crest, distance from contact point to alveolar crest and width of the papilla. The results showed that 6 months after the procedure, the percentage of change in the papilla level was higher in the test group (14,94% ± 21,35%) than in the control group (-1,39% ± 31,46%), though not statistically significant (p>0.05). There was no significant difference variation in the width of the papilla before and 4 months after the treatment in test group (p=0.09) and control group (p=0.16), and there was no significant difference variation in the distance between the tip of the papilla and the alveolar bone crest. There was significant improvement of the Pink Esthetic Score (PES) after 6 months in comparison to the initial condition in test group (p=0.0078; Wilcoxon), while there was no significant difference in the PES in control group 6 months after treatment (p=0.35). The results allow to conclude that the treatment for of the papilla deficiency using hyaluronic acid injection promotes improvement, similar to the results of the sub epithelial connective tissue graft treatment, but with significant esthetic improvement related specially to the color and texture characteristics of the adjacent soft tissues.

<p>Endopolyploid cells (hereafter - polyploid cells), which contain whole genome duplications in an otherwise diploid organism, play vital roles in development and physiology of diverse organs such as our heart and liver. Polyploidy is also observed with high frequency in many tumors, and division of such cells frequently creates aneuploidy (chromosomal imbalances), a hallmark of cancer. Despite its frequent occurrence and association with aneuploidy, little is known about the specific role that polyploidy plays in diverse contexts. Using a new model tissue, the Drosophila rectal papilla, we sought to uncover connections between polyploidy and aneuploidy during organ development. Our lab previously discovered that the papillar cells of the Drosophila hindgut undergo developmentally programmed polyploid cell divisions, and that these polyploid cell divisions are highly error-prone. Time-lapse studies of polyploid mitosis revealed that the papillar cells undergo a high percentage of tripolar anaphase, which causes extreme aneuploidy. Despite this massive chromosome imbalance, we found the tripolar daughter cells are viable and support normal organ development and function, suggesting acquiring extra genome sets enables a cell to tolerate the genomic alterations incurred by aneuploidy. We further extended these findings by seeking mechanisms by which the papillar cells tolerated this resultant aneuploidy.</p> / Dissertation

O uso de células tronco já é uma realidade em algumas áreas da Medicina, porém o mesmo não se aplica para a Odontologia, que segue utilizando materiais artificiais para substituir tecidos dentais perdidos. Desde 2000, quando Gronthos e colaboradores identificaram células-tronco na polpa de dentes permanentes, os estudos avançaram para que, num futuro breve, essas células possam ser de fato aplicadas para regenerar tecidos dentais, com destaque para a endodontia, onde o uso dessas células parece ser mais eminente. Dessa forma, esse trabalho procurou analisar a co-participação de células humanas transplantadas e células do hospedeiro em um modelo de engenharia pulpar. Para tanto, células de papila apical, enriquecidas ou não para o marcador CD146, foram transplantadas em câmaras pulpares despulpadas preenchidas com colágeno. Dois modelos animais foram utilizados, sendo um modelo transgênico para o gene GFP e outro imunocomprometido pela aplasia do timo (nude). Os resultados foram analisados nos dias 14 e 21 pós-transplante das amostras em cápsula renal. Nas amostras GFP realizou-se imunofluorescência para o marcador anti-GFP, com o objetivo de identificar as células do hospedeiro, enquanto nas amostras nude utilizou-se o marcador lâmina A para identificar as células humanas transplantadas. Nas análises morfológicas de todas as coroas transplantadas houve a formação de um tecido conjuntivo frouxo de celularidade variável, com a presença de vasos bem formados com eritrócitos em seu interior, inclusive nas coroas que receberam somente colágeno. Somente as amostras que receberam células houve a formação de matriz mineralizada no espaço pulpar, mas nos tempos experimentais analisados não foi possível visualizar as células humanas. Nas amostras nude, o marcador lâmina A foi negativo para todos os grupos que receberam transplante de células. Nas amostras GFPs, o marcador anti-GFP foi positivo na totalidade das células em todas as amostras estudadas. A partir disso concluiu-se que as células-tronco humanas de papila apical transplantadas apesar de terem desempenhado alguma função fisiológica, não foram identificadas após 14 e 21 dias e o tecido neoformado no interior da câmara pulpar era proveniente do hospedeiro. Adicionalmente, concluímos que não é necessário o transplante de células para a formação de um tecido conjuntivo frouxo no interior da câmara pulpar. / The use of stem cells is already a reality in some areas of Medicine, but the same does not apply for Dentistry, which keeps on using artificial materials to replace lost dental tissues. Since 2000, when Gronthos and coleagues identified stem cells in the pulp of permanent teeth, studies advanced so that, in the near future, these cells may actually be applied to regenerate dental tissues, especially in endodontics, in which the use of these cells seems to be more imminent. Thereby, this study sought to examine the co-participation of transplanted human cells and host cells in a model of dental pulp engineering. For this, apical papilla cells enriched or not with CD146 marker, were transplanted into decelluarized pulp chambers (empty crowns) filled with collagen. Two animal models were used, a transgenic model for the GFP gene and an immunocompromised by thymus aplasia (nude). The results were analyzed on days 14 and 21 after transplantation of samples into renal capsule. In the GFP samples immunofluorescence was performed for the anti-GFP marker in order to identify host cells, while in the nude samples the lamina A marker was used to identify the transplanted human cells. In the morphological analysis of all transplanted crowns there was formation of a loose connective tissue of variable cellularity, with the presence of well-formed vessels with erythrocytes inside, including in the crowns that received only collagen. Osteodentine was formed in the pulp chamber only in the samples that received cell, but after the wait time was not possible to visualize the human cells. In the nude samples the Lamina A marker was negative for all groups transplanted with cells. In the GFPs samples, the anti-GFP marker was positive for all cells in this group. We concluded that it was not necessary to use transplanted cells to form a connective tissue inside the pulp chamber and although transplanted human stem cells from apical papilla played some physiological function, after 14 and 21 days of transplantation, they were no longer present in the tissue newly formed by the host.

O uso de células tronco já é uma realidade em algumas áreas da Medicina, porém o mesmo não se aplica para a Odontologia, que segue utilizando materiais artificiais para substituir tecidos dentais perdidos. Desde 2000, quando Gronthos e colaboradores identificaram células-tronco na polpa de dentes permanentes, os estudos avançaram para que, num futuro breve, essas células possam ser de fato aplicadas para regenerar tecidos dentais, com destaque para a endodontia, onde o uso dessas células parece ser mais eminente. Dessa forma, esse trabalho procurou analisar a co-participação de células humanas transplantadas e células do hospedeiro em um modelo de engenharia pulpar. Para tanto, células de papila apical, enriquecidas ou não para o marcador CD146, foram transplantadas em câmaras pulpares despulpadas preenchidas com colágeno. Dois modelos animais foram utilizados, sendo um modelo transgênico para o gene GFP e outro imunocomprometido pela aplasia do timo (nude). Os resultados foram analisados nos dias 14 e 21 pós-transplante das amostras em cápsula renal. Nas amostras GFP realizou-se imunofluorescência para o marcador anti-GFP, com o objetivo de identificar as células do hospedeiro, enquanto nas amostras nude utilizou-se o marcador lâmina A para identificar as células humanas transplantadas. Nas análises morfológicas de todas as coroas transplantadas houve a formação de um tecido conjuntivo frouxo de celularidade variável, com a presença de vasos bem formados com eritrócitos em seu interior, inclusive nas coroas que receberam somente colágeno. Somente as amostras que receberam células houve a formação de matriz mineralizada no espaço pulpar, mas nos tempos experimentais analisados não foi possível visualizar as células humanas. Nas amostras nude, o marcador lâmina A foi negativo para todos os grupos que receberam transplante de células. Nas amostras GFPs, o marcador anti-GFP foi positivo na totalidade das células em todas as amostras estudadas. A partir disso concluiu-se que as células-tronco humanas de papila apical transplantadas apesar de terem desempenhado alguma função fisiológica, não foram identificadas após 14 e 21 dias e o tecido neoformado no interior da câmara pulpar era proveniente do hospedeiro. Adicionalmente, concluímos que não é necessário o transplante de células para a formação de um tecido conjuntivo frouxo no interior da câmara pulpar. / The use of stem cells is already a reality in some areas of Medicine, but the same does not apply for Dentistry, which keeps on using artificial materials to replace lost dental tissues. Since 2000, when Gronthos and coleagues identified stem cells in the pulp of permanent teeth, studies advanced so that, in the near future, these cells may actually be applied to regenerate dental tissues, especially in endodontics, in which the use of these cells seems to be more imminent. Thereby, this study sought to examine the co-participation of transplanted human cells and host cells in a model of dental pulp engineering. For this, apical papilla cells enriched or not with CD146 marker, were transplanted into decelluarized pulp chambers (empty crowns) filled with collagen. Two animal models were used, a transgenic model for the GFP gene and an immunocompromised by thymus aplasia (nude). The results were analyzed on days 14 and 21 after transplantation of samples into renal capsule. In the GFP samples immunofluorescence was performed for the anti-GFP marker in order to identify host cells, while in the nude samples the lamina A marker was used to identify the transplanted human cells. In the morphological analysis of all transplanted crowns there was formation of a loose connective tissue of variable cellularity, with the presence of well-formed vessels with erythrocytes inside, including in the crowns that received only collagen. Osteodentine was formed in the pulp chamber only in the samples that received cell, but after the wait time was not possible to visualize the human cells. In the nude samples the Lamina A marker was negative for all groups transplanted with cells. In the GFPs samples, the anti-GFP marker was positive for all cells in this group. We concluded that it was not necessary to use transplanted cells to form a connective tissue inside the pulp chamber and although transplanted human stem cells from apical papilla played some physiological function, after 14 and 21 days of transplantation, they were no longer present in the tissue newly formed by the host.

The aim of this study was to quantify the density of taste pores on the anterior region of the tongue, in adult males and 8 to 9 year old boys. Earlier studies had shown that, although 8 to 9 year olds were poorer than adults at sensing the tastant sucrose using a whole mouth procedure, localised regions of the tongue in male children were more sensitive than equivalent regions in adults. This study aims to detemine whether the age differences in sensitivity is related to a difference in taste pore density. Two areas of the tongue, for which children had previously been shown to have higher sensitivity than adults, were examined using a videomicrosocpy technique and the number and diameter of taste pores were measured. Children were found to have a greater density of taste pores, however the number of taste pores per papilla were similar in children and adults. It was found to be likely that the greater sensitivity of localised areas on the children's tongue is due to a greater taste pore density. The reduced sensitivity reported using whole mouth stimulation may be due to a reduced capacity to assimilate taste input from the whole mouth or due to different relative contributions to whole-mouth taste from the various receptive fields in the mouth. / Master of Science (Hons)

Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-10-05T17:26:27Z No. of bitstreams: 1 nathaliasaberdeandrade.pdf: 489327 bytes, checksum: 26e5218f3326304fb6e2eae34a98c1a5 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-10-09T19:58:47Z (GMT) No. of bitstreams: 1 nathaliasaberdeandrade.pdf: 489327 bytes, checksum: 26e5218f3326304fb6e2eae34a98c1a5 (MD5) / Made available in DSpace on 2017-10-09T19:58:47Z (GMT). No. of bitstreams: 1 nathaliasaberdeandrade.pdf: 489327 bytes, checksum: 26e5218f3326304fb6e2eae34a98c1a5 (MD5) Previous issue date: 2017-08-24 / Introdução: Esofagogastroduodenoscopia (EGD) convencional é o melhor método para avaliação do trato gastrointestinal superior (TGI), mas apresenta limitações para identificação da papila duodenal maior (PDM), mesmo após emprego da manobra de retificação. Exame completo da PDM está indicado nos pacientes de alto risco para adenocarcinoma da PDM. Objetivo: avaliar a utilização da pinça de biópsia (PB) durante EGD convencional como ferramenta adicional à manobra de retificação, na avaliação da PDM. Métodos: Foram estudados 671 pacientes entre 2013-2015 com busca ativa da PDM em três etapas: endoscópio não retificado, endoscópio retificado e uso da PB após retificação. Em todas elas registrou-se se: PDM foi totalmente visualizada (Posição A), parcialmente visualizada (Posição B) ou não visualizada (Posição C). Caso PDM não tenha sido completamente visualizada, paciente foi direcionado para etapa seguinte. Resultados: 341 eram sexo feminino (50,8%) com idade média de 49 anos. Dos 671 pacientes, em 324(48,3%) PDM foi identificada na posição A, 112(16,7%) em posição B e 235(35%) em posição C. Dos 347 pacientes submetidos à manobra de retificação, posição A foi encontrada em 186(53,6%), posição B em 51(14,7%) e posição C em 110(31,7%). Dos 161 pacientes restantes que utilizaram a PB, posição A foi vista em 94(58,4%), posição B em 14(8,7%) e posição C em 53(32,9%). A taxa acumulativa de visualização completa da PDM foi de 90%. Conclusão: O uso da PB aumentou a taxa de visualização completa da PDM em 14%, alcançando 604/671(90%) dos pacientes (P<0,01) avaliados e pode ser facilmente incorporada aos exames endoscópicos de rotina do TGI. / Introduction: Conventional esophagogastroduodenoscopy (EGD) is the best method for evaluation of the upper gastrointestinal tract (UGT), but it has limitations for the identification of the major duodenal papilla (MDP), even after the use of the straightening maneuver. Side-viewing duodenoscope is recommended for optimal examination of MDP in patients at high risk for lesions in this region. Objective: to evaluate the use of the biopsy forceps during conventional EGD as an additional tool to the straightening maneuver, in the evaluation of the MDP. Methods: A total of 671 patients were studied between 2013 and 2015, with active MDP search in three endoscope steps: not straightened, straightened and use of the biopsy forceps after straightening. In all of them it was recorded whether: MDP was fully visualized (Position A), partially visualized (Position B) or not visualized (Position C). If MDP was not fully visualized, patients continued to the next step. Results: 341 were female (50.8%) with mean age of 49 years. Of the 671 patients, 324 (48.3%) MDP was identified in position A, 112 (16.7%) in position B and 235 (35%) in position C. In the 347 patients who underwent the straightening maneuver, position A was found in 186 (53.6%), position B in 51 (14.7%) and position C in 110 (31.7%). Of the 161 remaining patients and after biopsy forceps use, position A was seen in 94 (58.4%), position B in 14 (8.7%) and position C in 53 (32.9%). The overall rate of complete visualization of MDP was 90%. Conclusion: the use of the biopsy forceps significantly increased the total MDP visualization rate by 14%, reaching 604/671 (90%) of the patients (P<0.01) and it can be easily incorporated into the routine endoscopic examination of the UGT.

In the amphibian auditory system, a subset of hair cells is known to be frequency tuned via electrical resonance. This tuning is thought to contribute to frequency selectivity of the information leaving the auditory periphery via the auditory afferent fibers. At the same time, while most, if not all, afferent fibers are shown to be frequency tuned, electrical resonance has only been experimentally demonstrated in a subset of amphibian auditory hair cells. In this thesis, we validate and use a novel Zap current method to probe the electrical resonance of the bullfrog amphibian papilla hair cells. We uncover the existence of two previously unknown types of electrically resonant auditory hair cells. We then show the existence of resonant hair cells across the length of amphibian papilla, with the range of frequency tuning that is nearly indistinguishable from that previously reported in the of auditory fibers. Therefore, this work further validates amphibian hair cell frequency resonance as the possible mechanism underlying frequency selectivity of the subsequent stages in auditory signal transduction.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第23094号 / 医博第4721号 / 新制||医||1050(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 髙橋 良輔, 教授 井上 治久, 教授 伊佐 正 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Hair disorders cause significant distress. The main, but limited, treatment for hair loss is minoxidil, an ATP-­sensitive potassium (KATP) channel opener whose mechanism of stimulation is unclear. The regulatory component of KATP channels has three forms: SUR1, SUR2A and SUR2B which all respond to different molecules. Minoxidil only opens SUR2B channels, though SUR1 and SUR2B are present in human hair follicles. To expand our understanding, the red deer hair follicle model was used initially. Deer follicles expressed the same KATP channel genes as human follicles when growing (anagen), but no channels were detected in resting follicles. This reinforces the importance of KATP channels in active hair growth and the usefulness of the deer model. To assess whether SUR1 KATP channels are actually involved in human hair growth, the effects of a selective SUR1 channel opener, NNC55-­9216, on scalp follicle growth in organ culture was examined. NNC55-­9216 stimulated anagen; its effect was augmented by minoxidil. This creates the potential for more effective pharmaceuticals to treat hair loss via SUR1 channels, either alone or in combination with minoxidil. The dermal papilla plays a crucial regulatory role in hair follicle activity determining the type of hair produced. Minoxidil had no effect on dermal papilla cell proliferation, but altered the profile of proteins produced when assessed by proteomics. Further research into the roles of KATP channels and greater understanding of the significance of these protein changes should enhance our knowledge of hair biology and help the development of new, improved therapies for hair pathologies.