Submitted by PPG Biologia Celular e Molecular (bcm@pucrs.br) on 2017-07-11T14:38:07Z No. of bitstreams: 1 NATALIA_ELTZ_SILVA_DIS.pdf: 1036979 bytes, checksum: 23cdf77b8842f9e9ade0b13e6f7bc111 (MD5) / Approved for entry into archive by Caroline Xavier (caroline.xavier@pucrs.br) on 2017-07-28T17:40:18Z (GMT) No. of bitstreams: 1 NATALIA_ELTZ_SILVA_DIS.pdf: 1036979 bytes, checksum: 23cdf77b8842f9e9ade0b13e6f7bc111 (MD5) / Made available in DSpace on 2017-07-28T17:47:24Z (GMT). No. of bitstreams: 1 NATALIA_ELTZ_SILVA_DIS.pdf: 1036979 bytes, checksum: 23cdf77b8842f9e9ade0b13e6f7bc111 (MD5) Previous issue date: 2017-03-08 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Aging-related diseases are becoming more common. Alzheimer's disease (AD), the most prevalent form of dementia, includes as initial symptoms cognitive deficits that are attributed to the toxic effects of amyloid? peptide (A?) that accumulates in senile plaques and neurofibrillary tangles composed of hyperphosphorylated tau protein. The amyloid cascade initially proposed to explain the effects of A? pointed to the plaques as the most toxic form of the A? molecule responsible for neuronal dysfunction and death. Recently, several evidences point to the increased toxicity of the soluble forms of the peptide. A better understanding of the dynamics of amyloid aggregation, clearance and toxic potential of the soluble versions may foster significant advances in the understanding AD mechanisms and the identification of potential targets for AD therapies. In this study we used the zebrafish as a model. 24-hour embryos received intracerebroventricular injection of human A?1-42 prepared to have different aggregation potentials: monomeric, oligomeric and plaqueforming. At 5 days post-fertilization (dpf), quantification of A?1-42 levels demonstrated a remnant increase in peptide levels in the animals injected with the solution that favored plaque formation. After monitoring for embryotoxic and teratogenic effects, 5dpf the animals were also evaluated in relation to general physiological aspects and their cognitive ability. Although the injection did not significantly impact animal survival or exploratory ability, the oligomeric solution induced specific cognitive deficits in relation to the vehicleinjected control. Together these results support the revised version of the amyloid cascade in which, although the presence of plaques corresponds to a greater accumulation of A?1- oligomeric forms may induce significant neurotoxic effects and result in cognitive deficits specially at disease?s early stages. / Com o envelhecimento da popula??o, doen?as relacionadas com o envelhecimento v?m se tornando mais comuns. A Doen?a de Alzheimer (DA), a forma prevalente de dem?ncia, inclui como sintomas iniciais deficit cognitivos que s?o atribu?dos a efeitos t?xicos de pept?deo ?- amiloide (A?) que se acumula em placas senis e emaranhados neurofibrilares constitu?dos pela prote?na tau hiperfosforilada. A cascata amiloide inicialmente proposta para explicar os efeitos do A? apontava para as placas de dep?sito de A?1-42 como a forma t?xica da mol?cula respons?vel pela disfun??o e morte neuronal. Recentemente diversas evid?ncias apontam para a toxicidade das vers?es sol?veis do pept?deo antes da agrega??o em placas. O melhor entendimento da din?mica de agrega??o do amiloide, limpeza e potencial t?xico das vers?es sol?veis pode permitir significativos avan?os no conhecimento dos mecanismos da doen?a e a identifica??o de potenciais alvos para terapias da DA. Neste estudo utilizamos o tele?steo zebrafish como modelo para a caracteriza??o destes processos. Embri?es com 24 horas receberam inje??o intracerebroventricular de A?1-42 humano preparado de forma a ter diferentes potenciais de agrega??o: monom?rica, oligom?rica e formadora de placas. Ao atingirem 5 dias p?s-fertiliza??o (dpf), a quantifica??o dos n?veis de A?1-42 demonstrou um aumento remanescente dos n?veis do pept?deo nos animais injetados com a solu??o que favorecia a forma??o de placas. Ap?s monitorarmos eventuais efeitos embriot?xicos e teratog?nicos, ao atingirem 5dpf, os animais foram tamb?m avaliados em rela??o a aspectos fisiol?gicos gerais e sua capacidade cognitiva. Embora a inje??o n?o tenha impactado significativamente a sobreviv?ncia dos animais ou a capacidade explorat?ria, a inje??o da solu??o oligom?rica induziu deficit cognitivos espec?ficos em rela??o ao controle injetado com ve?culo. Juntos, estes resultados suportam a vers?o revisada da cascata amiloide na qual, embora a presen?a de placas corresponda a um maior ac?mulo de A?1-42, a presen?a de vers?es oligom?ricas pode induzir efeitos neurot?xicos significativos e resultar em deficit cognitivos, especialmente nos est?gios iniciais da doen?a.

Submitted by PPG Medicina e Ci?ncias da Sa?de (medicina-pg@pucrs.br) on 2018-07-30T11:34:52Z No. of bitstreams: 1 GUSTAVO_DALTO_BARROSO_MACHADO.pdf: 989422 bytes, checksum: 22ea376b88186b8cd072559da6413b3b (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-08-02T14:55:19Z (GMT) No. of bitstreams: 1 GUSTAVO_DALTO_BARROSO_MACHADO.pdf: 989422 bytes, checksum: 22ea376b88186b8cd072559da6413b3b (MD5) / Made available in DSpace on 2018-08-02T15:07:57Z (GMT). No. of bitstreams: 1 GUSTAVO_DALTO_BARROSO_MACHADO.pdf: 989422 bytes, checksum: 22ea376b88186b8cd072559da6413b3b (MD5) Previous issue date: 2017-12-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The present study investigated if the model of memory dysfunction induced by iron overload, so far only tested in male rats, would be able to induce cognitive decline in female rats, aiming to validate it as an experimental model to study gender differences in neurodegenerative diseases. We also evaluated the effects of the activation of the G protein coupled estrogen receptor (GPER) on iron- and ovariectomy-induced memory deficits and the role of the PKA/CREB pathway as a mediator of its physiologic effects. Four experiments were performed: I ? female rats received iron (30mg/kg, per oral, p.o.) or vehicle (sorbitol, p.o.) in the neonatal period and after, in adulthood, they were submitted to the surgical protocol: ovariectomy (OVX) or false-surgery (SHAM), after 3 weeks, behavioral tasks were performed using the object recognition (RO); object location (OL) and inhibitory avoidance (IA) tasks; II ? the same two protocols cited before and behavioral tasks 3 weeks from the surgeries, but, immediately after the training sessions the animals received the selective agonist of the GPER, G1 (10mg/kg, subcutaneously, sc.) or vehicle (veh, oil, sc.), the long-term memory was assessed 24h after the training sessions; III ? dose-response curve in order to determine the the effects of the PKA-inhibitor (H-89) on memory retention of inhibitory avoidance task, the following doses were administered intraperitoneally to na?ve adult female rats: 0.1 mg/kg; 0.25mg/kg or 0.5mg/kg; and finally the experiment IV in which the memory of iron overloaded-ovariectomized female rats was assessed 24h after that they had received H-89 (0.25mg/kg, ip.) or veh 15 minutes previous to the training sessions and G1 (10mg/kg, sc.) or veh immediately after their training in the OL and IA tasks. We observed that the effects of iron overload in female, associated or non-associated to OVX varied according to the behavioral test that the animals were submitted. The lowest recognition index level in OL task was observed in the animals exposed to the iron overload and, subsequently, to the lack of estrogens secondary to ovariectomy. The acute post-training treatment with G-1 increased the recognition index in the OL task and it was also associated with an increasing in the latency to step down ifrom the platform to the grid in the IA task. Finally, all these positive results obtained with the post-training administration of G-1 were abolished when the animals received the pre-training treatment of H-89. Our study opens new avenues in the field of memory and estrogens once it introduces the association of lack of estradiol and iron overload as a pre-clinical phenotypical model of the aging process in women. Besides, from the best of our knowledge, it is the first evidence of the role of the G1 in the consolidation of the emotional memory as well as the first study connecting the GPER directly to the PKA/CREB pathway. / Este estudo investigou se o modelo de disfun??o cognitiva induzido pela sobrecarga de ferro, somente testado em ratos machos at? o momento, induz d?ficits de mem?ria em ratas, objetivando valid?-lo como um m?todo experimental que auxilie na compreens?o das diferen?as fisiopatol?gicas entre os sexos em rela??o ?s doen?as neurodegenerativas. Tamb?m avaliamos, de maneira in?dita, os efeitos da ativa??o receptor de estrog?nio acoplado ? prote?na G (GPER), nesse modelo combinado com o modelo de menopausa experimental. Al?m disso, especulamos o papel da via da PKA/CREB na ativa??o do GPER, atrav?s da inibi??o farmacol?gica da PKA. Foram realizados quatro experimentos para atingir esses objetivos: I ? ratas tratadas com ferro (via oral ? v.o.) ou ve?culo (sorbitol ? v.o.) foram ovariectomizadas ou submetidas ? cirurgia falsa (SHAM) na idade adulta, ap?s tr?s semanas do protocolo cir?rgico foram submetidas ?s tarefas de reconhecimento de objetos (RO), localiza??o de objetos (LO) e esquiva inibit?ria (EI); II ? ratas tratadas com ferro no per?odo neonatal foram ovariectomizadas ou submetidas ? cirurgia SHAM, ap?s recupera??o da cirurgia (3 semanas) foram tratadas anteriormente aos treinos das tarefas de LO e EI com G-1 (agonista seletivo do GPER, 10mg/kg, sc.) ou ve?culo (?leo de girassol, sc.) e testadas 24h ap?s o treino; III ? ratas naive receberam diferentes doses do composto H-89 (inibidor da PKA) para avaliar seu potencial amn?sico (0.1mg/kg, 0.25mg/kg e 0.5mg/kg; ip.) e IV ? ratas submetidas ? sobrecarga neonatal de ferro e OVX, ap?s 3 semanas do protocolo cir?rgico receberam quinze minutos antes dos treinos das tarefas de LO e EI o composto H-89 (0.25mg/kg, ip.) ou ve?culo (salina, ip.) e imediatamente ap?s o treino G-1 (10mg/kg, sc.) ou ve?culo (?leo de girassol, sc.), as ratas foram testadas ap?s 24h dos treinos. Os efeitos do protocolo de sobrecarga neonatal de ferro associado ou n?o a priva??o de estradiol variou em depend?ncia da tarefa comportamental avaliada. Na tarefa de LO os grupos submetidos ? sobrecarga neonatal de ferro e ovariectomia apresentaram os menores ?ndices de reconhecimento. O tratamento com o agonista do GPER ? G1 - foi capaz de melhorar os d?ficits cognitivos induzidos pela sobrecarga neonatal de ferro e/ou pela ovariectomia em ratas no teste de LO e EI, esses efeitos foram inibidos pela administra??o do inibidor da PKA anteriormente ao treino das tarefas. Demonstramos que o modelo de sobrecarga neonatal de ferro associado ? ovariectomia ? um modelo ?til para o estudo dos efeitos delet?rios da diminui??o dos n?veis s?ricos de estradiol na menopausa, auxiliando como modelo fenot?pico do que ocorre na cl?nica, onde as mulheres p?s-menopausa apresentam preval?ncia maior de Doen?a de Alzheimer que homens. Al?m disso, demonstramos de maneira in?dita os efeitos do composto G-1 na tarefa de esquiva inibit?ria. Nossos dados comportamentais tamb?m sugerem que o composto G-1 depende da ativa??o da PKA para exercer seus efeitos sobre a consolida??o da mem?ria.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-03-22T00:26:40Z No. of bitstreams: 1 MariaDeFatimaBezerra_TESE.pdf: 8001077 bytes, checksum: 680617b92afca9b7140a3d99473e34c2 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-03-22T23:44:28Z (GMT) No. of bitstreams: 1 MariaDeFatimaBezerra_TESE.pdf: 8001077 bytes, checksum: 680617b92afca9b7140a3d99473e34c2 (MD5) / Made available in DSpace on 2016-03-22T23:44:28Z (GMT). No. of bitstreams: 1 MariaDeFatimaBezerra_TESE.pdf: 8001077 bytes, checksum: 680617b92afca9b7140a3d99473e34c2 (MD5) Previous issue date: 2015-04-24 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / A presente tese avaliou a polpa de jambol?o (Eugenia jambolana Lam.) fresca e desidratada mediante dois processos de secagem (liofiliza??o e atomiza??o com 10 % de goma Ar?bica) quanto as caracter?sticas f?sico-qu?micas (pH, umidade, atividade de ?gua, di?metro m?dio de part?culas, solubilidade dos p?s e cor instrumental), bioativas [compostos fen?licos totais (CFT), antocianinas monom?ricas, proantocianidinas (PA), ?cido el?gico total (AET), miricetina e cianidina] e estudo da funcionalidade in vitro (atividade antioxidante, antienzim?tica e antimicrobiana). Em seguida, a funcionalidade in vivo da polpa de jambol?o foi avaliada, com uso do modelo Caenorhabditis elegans, quanto ? via sinaliza??o de insulina, longevidade e doen?as neurodegenerativas (doen?a de Alzheimer e Mal de Parkinson). A polpa de jambol?o desidratada apresentou consider?vel reten??o de CFT (50 % a 75 %), PA (90 % a 98 %), AET (31 % a 83 %), miricetina (40 % a 84 %), cianidina (72 % a 84 %) e atividade antioxidante (15 %). A polpa fresca, o p? liofilizado e o p? atomizado apresentaram elevada atividade inibit?ria contra lipase pancre?tica (4,4 a 5,8 mg/mL), alfa-glicosidase (10,3 a 13,8 mg/mL) e alfa-amilase (8,9 a 11,2 mg/mL). Esses grupos experimentais tamb?m se apresentaram inibidores ativos contra o crescimento de S. aureus. Os p?s de jambol?o aumentaram a express?o de genes ligados ? via de sinaliza??o de insulina (SIR-2.1, PPTR-1, DAF-16, SOD-3, e CTL) e foram capazes de estender o tempo m?dio de vida de C. elegans (18,07 % a 24,34 %), reduzir a paralisia induzida pelo amiloide AB1-42 e os danos causados pela mol?cula neurot?xica 1-methyl-4- phenylpyridinium (MPP+ ). A partir disso, foi desenvolvido frozen yogurt com uso do leite caprino adicionado de Bifidobacterium animalis subsp. lactis BI-07 com polpa ou p? atomizado de jambol?o. O produto final foi avaliado quanto a suas caracter?sticas f?sico-qu?micas (pH, acidez titul?vel, s?lidos totais, prote?na, a??cares redutores totais, gordura, cinzas, overrun e teste de derretimento], bioativas (CFT e antocianinas monom?ricas), atividade antioxidante, iii viabilidade da cultura probi?tica e an?lise sensorial (teste de aceita??o). As amostras de frozen yogurt caprino com cultura probi?tica apresentaram menor pH e maior acidez, CFT, antocianinas e atividade antioxidante quando comparada com aquelas sem a presen?a da B. animalis. Foram observados n?veis de overrun entre 14,2% e 22,6%. As amostras de frozen yogurt com cultura probi?tica alcan?aram resultados inferiores para o atributo sabor. De maneira geral, a presente pesquisa apresenta o jambol?o como um fruto rico em compostos bioativos, com elevada capacidade funcional, com potencial para modular importantes vias biol?gicas, aumentar a expectativa de vida e retardar risco de doen?as neurodegenerativas. Atualmente o jambol?o ? um fruto ex?tico subaproveitado no Brasil com elevado poder corante e a presente tese mostra, pela primeira vez na literatura, importantes achados tecnol?gicos, biol?gicos e cient?ficos sobre essa fruta que podem ser usados para o desenvolvimento de produtos alimentares saud?veis. / This work evaluated the fresh, spray dried (with 10 % of Arabic Gum) and freeze dried jambolan pulp (Eugenia jambolana Lam.) in regard to physicochemical (pH, moisture, water activity, average particle diameter, solubility and color), bioactive [total phenolic content (TPC), monomeric anthocyanin, pronathocyanidin (PA), total elagic acid (TEA), myricetin and cyanidin] and in vitro functionality (antioxidant, antienzymatic and antimicrobial activities]. In addition, the in vivo functionality of jambolan pulp was investigated using the Caenorhabditis elegans model for insulin signaling, longevity and induced neurodegeneration (Alzheimer?s disease and Parkinson?s disease related symptoms). The dried jambolan pulp presented TPC retention (50% to 75%), PA (90% to 98%), TEA (31% to 83%), myricetin (40% to 84%), cyanidin (72% to 84%) and antioxidant activity (15%). The fresh jambolan pulp, the freeze dried pulp and the spray dried jambolan pulp presented high enzymatic inhibitory activity against pancreatic lipase (4,4 to 5,8 mg/mL), alpha-glycosidase (10,3 to 13,8 mg/mL) and alpha-amylase (8,9 to 11,2 mg/mL). They also were active inhibitors against the pathogen S. aureus. The dried jambolan experimental samples were able to increase the expression of several genes linked to the insulin signaling pathways (SIR-2.1, PPTR-1, DAF-16, SOD-3, e CTL) and increased the lifespan in C. elegans (18,07 % - 24,34 %), besides decreasing the amyloid AB1-42 aggregation induced paralysis and MPP+ (1-methyl-4-phenylpyridinium) induced neurodegeneration. Based on that, the jambolan pulp and the spray dried jambolan pulp were further selected for the production of caprine frozen yogurt with the addition of Bifidobacterium animalis subsp. lactis BI-07. The final product were evaluated in regard to their physicochemical (pH, acidity, total solids, protein, total reducing sugars, fat, ashes, overrun, melting test), bioactive (TPC and monomeric anthocyanin, antioxidant activity, probiotic viability and sensory analysis (sensory acceptance). The results showed that samples with probiotic had lowest pH and higher acidity, TPC, anthocyanin and antioxidant activity. It was also observed low overrun (14.2% to 22.6%). vi Samples with probiotic had lower flavor scores. Overall, this research presents the jambolan as a highly functional bioactive-rich fruit with the potential to modulate important biological pathways, extend lifespan and retard the development of neurodegenerative diseases. Jambolan is an underexploited exotic fruit with a high colorant potential and this thesis shows for the first time in the literature important technological, biological and scientific data about this fruit that could be used towards the development of health-oriented food products.

Made available in DSpace on 2014-12-17T14:03:36Z (GMT). No. of bitstreams: 1 JullianeTAM_TESE_Parcial.pdf: 7138229 bytes, checksum: a06700fccf67cc2db086f62bf86db506 (MD5) Previous issue date: 2014-02-26 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / studies using UV as a source of DNA damage. However, even though unrepaired UV-induced DNA damages are related to mutagenesis, cell death and tumorigenesis, they do not explain phenotypes such as neurodegeneration and internal tumors observed in patients with syndromes like Xeroderma Pigmentosum (XP) and Cockayne Syndrome (CS) that are associated with NER deficiency. Recent evidences point to a role of NER in the repair of 8-oxodG, a typical substrate of Base Excision Repair (BER). Since deficiencies in BER result in genomic instability, neurodegenerative diseases and cancer, it was investigated in this research the impact of XPC deficiency on BER functions in human cells. It was analyzed both the expression and the cellular localization of APE1, OGG1 e PARP-1, the mainly BER enzymes, in different NER-deficient human fibroblasts. The endogenous levels of these enzymes are reduced in XPC deficient cells. Surprisingly, XP-C fibroblasts were more resistant to oxidative agents than the other NER deficient fibroblasts, despite presenting the highest of 8-oxodG. Furthermore, subtle changes in the nuclear and mitochondrial localization of APE1 were detected in XP-C fibroblasts. To confirm the impact of XPC deficiency in the regulation of APE1 and OGG1 expression and activity, we constructed a XPC-complemented cell line. Although the XPC complementation was only partial, we found that XPC-complemented cells presented increased levels of OGG1 than XPC-deficient cells. The extracts from XPC-complemented cells also presented an elevated OGG1 enzimatic activity. However, it was not observed changes in APE1 expression and activity in the XPCcomplemented cells. In addition, we found that full-length APE1 (37 kDa) and OGG1- ? are in the mitochondria of XPC-deficient fibroblasts and XPC-complemented fibroblasts before and after induction of oxidative stress. On the other hand, the expression of APE1 and PARP-1 are not altered in brain and liver of XPC knockout mice. However, XPC deficiency changed the APE1 localization in hypoccampus and hypothalamus. We also observed a physical interaction between XPC and APE1 proteins in human cells. In conclusion, the data suggest that XPC protein has a role in the regulation of OGG1 expression and activity in human cells and is involved mainly in the regulation of APE1 localization in mice. Aditionally, the response of NER deficient cells under oxidative stress may not be only associated to the NER deficiency per se, but it may include the new functions of NER enzymes in regulation of expression and cell localization of BER proteins / A maior parte do nosso conhecimento sobre a via de Reparo de Excis?o Nucleot?deos (NER) vem de estudos usando a luz ultravioleta (UV) como fonte de danos no DNA. Contudo, embora os danos no DNA causados pela luz UV sejam relacionados ? ocorr?ncia de mutag?nese, morte celular e tumorig?nese, eles n?o justificam fen?tipos como neurodegenera??o e tumorig?nese observados em pacientes com s?ndromes como Xeroderma Pigmentosum (XP) e S?ndrome de Cockayne (CS), as quais s?o associadas ? defici?ncia na via NER. Adicionalmente, evid?ncias mais recentes indicam o envolvimento da via NER no reparo de 8-oxodG, um substrato t?pico da via de Reparo por Excis?o de Bases (BER). Uma vez que a defici?ncia na via BER resulta em instabilidade gen?mica, doen?as neurodegenerativas e c?ncer, foi investigado neste trabalho o impacto da defici?ncia em XPC nas fun??es da via BER em c?lulas humanas. Foram realizadas an?lises da express?o e da localiza??o celular de APE1, OGG1 e PARP-1, principais enzimas da via BER, em fibroblastos humanos deficientes na via NER. Os resultados demonstraram que os n?veis end?genos de APE1, PARP-1 e OGG1 s?o reduzidos nos fibroblastos deficientes em XPC, os quais foram mais resistentes a diferentes tipos de agentes oxidantes e apresentaram n?veis elevados de 8-oxodG quando comparados aos demais fibroblastos deficientes na via NER. Adicionalmente, altera??es sutis na localiza??o nuclear e mitocondrial de APE1 foram observadas nos fibroblastos deficientes em XPC. Para confirmar o impacto da defici?ncia de XPC na regula??o da express?o e atividade de APE1 e OGG1, foi constru?da uma linhagem complementada com XPC. Embora a complementa??o tenha sido parcial, foi poss?vel observar que os fibroblastos parcialmente complementados com XPC apresentaram n?veis maiores de express?o de OGG1 quando comparados aos fibroblastos deficientes em XPC. Os extratos dos fibroblastos parcialmente complementados com XPC tamb?m apresentaram uma elevada atividade enzim?tica de OGG1. Contudo, n?o foram observadas mudan?as na express?o e atividade de APE1 nos fibroblastos parcialmente complementados com XPC. Adicionalmente, foi poss?vel verificar a presen?a da forma completa de APE1 (37 kDa) e de OGG1-? na mitoc?ndria dos fibroblastos deficientes em XPC e parcialmente complementados com XPC. Por outro lado, observou-se que a express?o de APE1 e PARP-1 n?o ? alterada no c?rebro e f?gado de camundongos knockouts para XPC. Contudo, a defici?ncia em XPC resultou em mudan?as na localiza??o celular de APE1 no hipocampo e hipot?lamo. Ainda, foi observada a ocorr?ncia de uma intera??o f?sica entre as prote?nas XPC e APE1 em c?lulas humanas. Em conclus?o, os dados sugerem que a prote?na XPC possui um papel na regula??o da express?o e da atividade de OGG1 em c?lulas humanas e est? envolvida na regula??o da localiza??o celular de APE1 principalmente em camundongos. Adicionalmente, as respostas celulares dos fibroblastos deficientes na via NER ao estresse oxidativo podem n?o estar associadas ? defici?ncia na via NER per se, mas podem incluir novas fun??es das enzimas da via NER na regula??o da express?o e localiza??o celular das prote?nas da via BER / 2020-01-01