Chemotherapy is one of the most important treatments for cancer. However, systemic toxicity, drug resistance and unstable kinetics of the drug in the blood are serious problems of chemotherapy. The use of biodegradable polymers for controlled release of anticancer drugs has gained popularity in recent years. Controlled release of drugs from polymeric carriers has some advantages such as improvement in the efficiency of treatment, reduction in systemic toxicity and prevention of the drug resistance that is developed by the cancer cells. In this study, poly(D,L-lactide-co-glycolide) microparticles were used as carriers for the controlled release of all-trans-Retinoic acid, tamoxifen, tamoxifen citrate and idarubicin. It was aimed to prepare a drug carrier system for controlled release of hydrophobic anticancer drugs. The empty and drug loaded poly (D,L-lactide-co-glycolide) microparticles were prepared by solvent extraction/evaporation technique with single emulsion (oil/water). Optimized microparticles were characterized by using inverted light microscopy and scanning electron microscopy to examine their morphology and sizes. Drug content of microparticles and the amount of released drug were determined spectrophotometrically. In vitro toxicity of the microparticles on MCF-7 human breast cancer cells was investigated. It was revealed that the microparticles were smooth and spherical in shape. Their sizes differed in the range of 2-20 &micro / m. atRA-loaded microparticles showed approximately 90% encapsulation efficiency and it was confirmed that changing in drug/polymer ratio affected the extend of drug content. Increase in drug content caused a slower release pattern. Moreover, although the empty microparticles caused some toxicity, atRA-loaded PLGA microparticles showed slight cell growth inhibition.

Retinol (vitamin A) and its active metabolite, all-trans retinoic acid, signal through nuclear retinoic acid and retinoid X receptor (RAR/RXR) heterodimers. These complexes regulate the expression of genes involved in developmental processes such as limb development. In excess, retinoids are potent teratogens and cause marked reductive effects on the developing limb. The goal of this thesis was to elucidate the molecular mechanisms underlying retinoid-induced limb dysmorphogenesis. Specifically, using an in vitro limb culture system, I examined the involvement of one RAR isoform, RARgamma, in mediating retinoid insult. / My first objective was to examine how limbs deficient in RARgamma responded to exogenous retinoid exposure. I showed that RARgamma-null limbs (on an RARalpha1-null background) exhibited less severe limb defects following retinoid insult when compared to their wild-type counterparts. Additionally, the absence of RARgamma abolished the retinoid-induced misregulation of genes important for chondrogenesis (Sox9 and Col2a1 ) and limb outgrowth (Meis-1 and -2). / The next objective set out to determine how pharmacological activation of RARgamma affected limb development. The RARgamma-selective agonist (BMS-189961) caused limb dysmorphology (namely, effects on cartilage) that was comparable to pan-RAR activation with all-trans retinoic acid. A chondrogenesis-focused gene array analysis identified Mgp and Gdf10 as two RARgamma-responsive genes that may mediate retinoid-induced limb insult. / Subsequently, I assessed the functional involvement of Mgp in mediating retinoid teratogenicity. Limbs were treated with all- trans retinoic acid and warfarin (an inhibitor of MGP); warfarin co-treatment rescued limbs from retinoid-induced insult. / My final objective was to determine the importance of Gdf10 in mediating limb development. Recombinant human Gdf10-soaked beads were implanted into distal limb structures; ectopic overexpression of Gdf10 in the web (but not the digital ray) resulted in marked proximal limb malformations. / Collectively, these studies have illustrated the importance of RARgamma in retinoid teratology and have identified several potential mechanisms by which retinoids cause limb defects.

Les rétinoïdes sont utilisés dans le traitement d’une variété de tumeurs malignes et lésions précancéreuses. Leurs effets dans des lignées cellulaires dérivées de tumeurs solides tel que le cancer du sein ont été étudiés extensivement. Cependant, les bénéfices dans le cancer du sein restent à date peu clairs. Ceci est probablement du à l’hétérogénéité des tumeurs mammaires et la réponse très variable aux effets antiprolifératifs de l’acide rétinoïque. Dans les lignées cellulaires cancéreuses mammaires, la réponse l’AR est fortement corrélée au niveau d’expression du récepteur aux estrogènes alpha (ERα), qui régule l’expression du gène qui encode le récepteur à l’acide rétinoïque alpha, RARA. Malgré cela, certaines lignées cellulaires ER-négatives, comme la lignée HER2-positive SK-BR-3, ont été décrites comme étant sensibles à l’AR. Dans le Chapter 2: de cette thèse, nous avons étudié les mécanismes de la signalisation ER-dépendante et ER-indépendante dans les cellules cancéreuses mammaires. Nous avons utilisé des lignées ER-négatives et ER-positives pour démontrer qu’une partie de la réponse à l’AR est indépendante de la signalisation par ER. Nous avons identifié plusieurs gènes cibles primaires de l’AR qui ont des effets similaires à l’AR quand ils sont surexprimés dans des cellules mammaires cancéreuses. Cette étude apporte une meilleure compréhension des mécanismes complexes qui mènent à l’arrêt de croissance induit par l’AR dans les cellules cancéreuses mammaires. Dans le Chapitre 3, nous avons regardé plus en détails la signalisation ER-indépendante par l’AR dans des cellules ayant une amplification des gènes HER2 et RARA et nous avons identifié une synergie entre l’AR et le Herceptin dans ces cellules. Nous proposons que les gènes FOXO jouent une rôle dans cette synergie. Les cellules SK BR 3, ayant une coamplification HER2/RARA, pourraient représenter une classe de tumeurs qui pourraient bénéficier d’un traitement avec des rétinoïdes, en augmentent la réponse au Herceptin et potentiellement en réduisant la résistance au Herceptin. En conclusion, les données présentées dans cette thèse aident à mieux comprendre les mécanismes menant à l’arrêt de croissance induit par l’AR dans les cellules cancéreuses mammaires et fournissent une application potentielle pour l’utilisation de l’AR dans le traitement du cancer du sein. / Retinoids are being used in the treatment of several malignancies and precancerous lesions. Their effects on cell lines derived from solid tumors, such as breast cancer, have also been described extensively. Their benefit in breast cancer, however, remains unclear. This might be because of the high levels of heterogeneity of breast tumors and the very variable response to the antiproliferative effects of retinoic acid. In mammary tumor cell lines, the response to retinoic acid is highly correlated with the expression of the estrogen receptor alpha (ERα), which regulates the expression of the retinoic acid receptor alpha gene RARA. However, some ER-negative cell lines, such as the HER2 positive SK-BR-3 cell line, have been reported to be RA-sensitive. In Chapter 2: of this thesis we have investigated the mechanisms of ER-dependent and ER-independent RA signaling in breast cancer cells. Using ER-positive and ER-negative cell lines, we show that part of the response to RA is independent of ER signaling. Several direct retinoic acid targets were identified that could mimic antiproliferative effects of retinoic acid when overexpressed in breast cancer cells. This study has provided better insight in the complex mechanisms that lead to RA-induced growth arrest in breast cancer cells. In Chapter 3: we looked further into the ER-independent RA signaling in HER2/RARA-amplified cells and identified a synergy between RA and Herceptin in these cells. We propose a role for FOXOs in mediating this synergy. HER2/RARA coamplified breast tumors might represent a subclass of tumors that could benefit from retinoid treatment, both increase antitumor effects of Herceptin, as well as in potentially reducing Herceptin resistance. In conclusion, data presented in this thesis give better insight in the mechanisms of RA induced growth arrest in breast cancer cells and provide a potential application of retinoids in a subset of breast tumors.

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (p. 133-145).

Cancer is a well-known disease that many people encounter in their lifetime. There is constantly research being performed on cancer to find treatments for those types where none has been found, or even find better or more efficient treatments for those cancer types where there already is treatment available. Two types of cancer that have been studied in this thesis are neuroblastoma, which is a form of cancer that affects children and infants, and breast cancer. The 13-cis retinoic acid is presently used as treatment for neuroblastoma post-surgery and post-chemo therapy, but the treatment is quite invasive. It has been shown that 1,25-dihydroxyvitamin D3 is a good candidate for cancer treatment, and the aim of this study was to investigate whether a combination of 1,25-dihydroxyvitamin D3 and two forms of retinoic acid, all-trans and 13-cis, could cause synergistic effects on cell viability, invasion, and differentiation of the cells. The two vitamins were combined at different concentrations and ratios to make the different treatments. A proliferation assay with absorbance measurement was performed to determine cell viability, and a migration assay was performed to observe the migratory properties of the cells after treatment. The results concluded that the combined treatments had greater effect than the single treatments on cell viability in both neuroblastoma and breast cancer cells. The results showed that single treatment of 13-cis retinoic acid and combined treatments had the highest effect on invasion and differentiation on neuroblastoma cells.

Data de aprova??o ausente. / Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2016-12-20T12:32:00Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) leticia_marcia_silva_tinoco.pdf: 1813667 bytes, checksum: cc4f9146534721910e6a3dde4429645b (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2017-01-13T15:55:44Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) leticia_marcia_silva_tinoco.pdf: 1813667 bytes, checksum: cc4f9146534721910e6a3dde4429645b (MD5) / Made available in DSpace on 2017-01-13T15:55:44Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) leticia_marcia_silva_tinoco.pdf: 1813667 bytes, checksum: cc4f9146534721910e6a3dde4429645b (MD5) Previous issue date: 2016 / O ?cido retinoico (AR), um derivado da vitamina A, ? um dos exemplos mais bem-sucedidos de f?rmacos usados na terapia de diferencia??o do c?ncer. O AR ? um f?rmaco altamente lipof?lico (log P 4,6) que apresenta baixa solubilidade aquosa, limitando sua utiliza??o parenteral. Dessa forma, sua incorpora??o em nanocarreadores lip?dicos tem sido proposta como uma alternativa promissora para a administra??o desse f?rmaco. O maior direcionamento dos nanossistemas para as c?lulas tumorais pode ser obtido por meio de modifica??es na superf?cie deste, como o revestimento com ?cido hialur?nico (AH), que se liga a receptor CD44 sobre-expresso em alguns tumores. Assim sendo, este trabalho teve por objetivo principal desenvolver, caracterizar e avaliar a atividade antineopl?sica in vitro de nanoemuls?es contendo AR revestidas e n?o revestidas com AH para tratamento de c?ncer de mama. Primeiramente, foi desenvolvido um m?todo espectrofotom?trico, com detec??o do AR em 324 nm, que foi validado em rela??o ? seletividade em rela??o aos componentes de matriz e aos produtos de degrada??o for?ada, linearidade, precis?o, exatid?o e robustez, conforme a legisla??o pertinente. Em seguida, foram desenvolvidas e caracterizadas nanoemuls?es (NE) preparadas por emulsifica??o espont?nea e revestidas eletrostaticamente com AH em diferentes concentra??es. Ap?s o revestimento com 0,5 mg/mL de AH, a NE escolhida apresentou tamanho de 158 ? 5 nm, distribui??o monodispersa e potencial zeta de -19,7 ? 1,20 mV e um teor de encapsula??o de 99,2 ? 0,5%, estando compat?vel com a administra??o parenteral. Al?m disso, as formula??es revestidas permaneceram est?veis ao longo de 60 dias armazenadas a 4?C. O perfil de libera??o do AR a partir dos nanossistemas foi avaliado e observou-se que segue uma cin?tica de primeira ordem, de forma que a taxa de libera??o depende da concentra??o do f?rmaco ainda presente na matriz. Foram, por fim, avaliados os efeitos citot?xicos em linhagens de c?lulas de c?ncer de mama (MCF-7 e MDA-MB-231) e fibroblastos normais (L929) e observou-se que a formula??o revestida promove aumento de atividade antic?ncer do AR, especialmente nas c?lulas que expressam mais os receptores CD44 (MDA-MB-231) e reduzida toxicidade em rela??o ?s c?lulas normais. Desta forma, a encapsula??o do AR em NE revestida por AH pode ser uma abordagem interessante para aumentar a efic?cia e a biodisponibilidade do AR no tratamento do c?ncer de mama e outros tipos de c?ncer que sobre-expressam esse receptor. / Disserta??o (Mestrado) ? Programa de P?s-gradua??o em Ci?ncias Farmac?uticas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2016. / Retinoids, such as all-trans retinoic acid (RA), are structural molecules derived from vitamin A and play important roles in various cell types, especially on vision, cell proliferation and differentiation. RA is a lipophilic acid (log P 4.6) with low aqueous solubility, which limits its pharmaceutical use. In order to allow its parenteral administration, an interesting alternative could be its incorporation in lipid nanocarrier systems, such as nanoemulsions (NE). On the other hand, it is possible to increase the target to CD44 receptor expressing tumor cells, coating the NE surface with hyaluronic acid (HA). Therefore, the main goal of this study was to develop, characterize and evaluate the in vitro antitumor activity of RA-loaded NE coated with HA for the treatment of breast cancer. First, a spectrophotometric method was developed, with RA detection at 324 nm, which was validated with regard to selectivity to matrix components and forced degradation products, linearity, precision, accuracy and robustness, in accordance with the proper legislation. Then, the NE, prepared by the spontaneous emulsification and electrostatically coated with HA in different concentrations, were developed and characterized. After coating with 0.5 mg/mL, the chosen HA-coated NE presented size of 158 ? 5 nm, monodisperse distribution, zeta potential of -19.7 ? 1.20 mV and encapsulating efficiency of 99.2 ? 0.5%, which is compatible with parenteral administration. Moreover, the coated formulations remained stable over 60 days stored at 4 ?C. The RA release profile from the nanosystems was evaluated and it follows the first order kinetics, so that the release rate depends on the drug concentration still present in the matrix. Finally, the cytotoxic effects were assessed in breast cancer cell lines (MCF-7 and MDA-MB-231) and in normal fibroblasts (L929), and found that the coated formulation promotes increased anticancer activity of RA, especially in cells expressing plus CD44 receptors (MDA-MB-231) and reduced toxicity compared to normal cells. Thus, the encapsulation of RA in HA-coated NE can be an interesting approach to increase the RA efficacy and bioavailability in the treatment of breast cancer and other cancers overexpressing that receptor.

Submitted by Camila Girondi (camila.girondi@gmail.com) on 2018-07-03T20:30:17Z No. of bitstreams: 1 Dissertação 28.06 COM FICHA CATALOGRÀFICA.pdf: 1652288 bytes, checksum: 2ff0f9722f6563cc30aaacbdd2bd4b81 (MD5) / Approved for entry into archive by Silvana Alvarez null (silvana@ict.unesp.br) on 2018-07-10T19:06:46Z (GMT) No. of bitstreams: 1 girondi_cm_me_sjc.pdf: 1652288 bytes, checksum: 2ff0f9722f6563cc30aaacbdd2bd4b81 (MD5) / Made available in DSpace on 2018-07-10T19:06:46Z (GMT). No. of bitstreams: 1 girondi_cm_me_sjc.pdf: 1652288 bytes, checksum: 2ff0f9722f6563cc30aaacbdd2bd4b81 (MD5) Previous issue date: 2018-05-02 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / As nanofibras (NF) podem ser sintetizadas por meio de eletrofiação, um método simples que permite incorporação de fármacos em biopolímeros que apresentam a vantagem de serem liberados de forma gradual. Policaprolactona (PCL) apresenta a vantagem de poder ser eletrofiada permitindo a incorporação de fármacos. Os ácidos retinoicos são fármacos empregados no tratamento de osteossarcoma, mas, apresentam efeitos colaterais. O ATRA (all-trans-retinoic-acid) é o mais comum entre os retinoides. Os objetivos deste estudo foram: sintetizar e caracterizar NF, incorporar o fármaco a elas e verificar a liberação ao meio de cultura; realizar testes de citoxicidade e genotoxicidade tais como: crescimento, integridade, viabilidade e micronúcleo de células MG63 tratadas com NF incorporada com ATRA e analisar a secreção das citocinas pró inflamatórias: IL-1β, IL-6 e TNF-α por meio do teste ELISA. As NF sintetizadas por meio da eletrofiação foram caracterizadas morfologicamente e bioquimicamente por meio de microscopia eletrônica de varredura (MEV) e estudo do espectro de infravermelho da transformada de Fourrier (FTIR): Os testes in vitro foram realizados utilizando uma linhagem de células imortalizadas de osteossarcoma, MG63. Os ensaios incluídos nesse estudo foram divididos em grupos compostos por: [C (controle puro), ATRA (controle positivo), NF (controle negativo) e NF + ATRA (grupo experimental).] Em todos os grupos foram realizados testes de viabilidade celular (MTT), micronúcleo e ELISA sob os tempos de 3, 7 e 14 dias. A liberação do fármaco através do sistema de NF+ATRA foi comprovada pelo FTIR. Os demais resultados encontrados foram avaliados através do teste ANOVA one-way, no qual o método apresentou-se eficiente, mostrando diferença estatística de p=0,00, sendo signficante para todos os períodos de tempo nos testes de MTT e MN, além de ser verificada a secreção de citocinas proinflamatórias IL-1β e IL-6, comprovando os efeitos citotóxicos e genotoxicos sob células MG63 mostrando a efetividade da técnica. Podendo ser uma alternativa para o tratamento. / Nanofibers (NF) can be synthesized by electro-spinning, a simple method that allows incorporation of drugs into biopolymers that have the advantage of being released gradually. Polycaprolactone (PCL) has the advantage that it can be electrophied allowing the incorporation of drugs. Retinoic acids are drugs used in the treatment of osteosarcoma, but have side effects. ATRA (all-trans-retinoicacid) is the most common retinoid. The objectives of this study were: to synthesize and characterize NF, to incorporate the drug to them and to verify the release to the culture medium; perform cytotoxicity and genotoxicity tests such as: growth, integrity, viability and micronucleus of N63-treated NF-treated cells with ATRA and to analyze the secretion of pro-inflammatory cytokines: IL-1β, IL-6 and TNFα by the ELISA . The NF synthesized by electrophilation were characterized morphologically and biochemically by scanning electron microscopy (SEM) and Fourrier Transform Infrared Spectrum (FTIR) study: In vitro tests were performed using an immortalized osteosarcoma cell line, MG63. The trials included in this study were divided into groups composed of: C (pure control), ATRA (positive control), NF (negative control) and NF + ATRA (experimental group). All groups were tested for cell viability (MTT), micronucleus and ELISA under the times of 3, 7 and 14 days. Release of the drug through the NF + ATRA system was confirmed by FTIR. The other results were evaluated using the oneway ANOVA test, in which the method was efficient, showing a statistical difference of p = 0.00, being significant for all time periods in the MTT and MN tests, besides the secretion of proinflammatory cytokines IL-1β and IL-6, proving the cytotoxic and genotoxic effects under MG63 cells, showing the effectiveness of the technique. It may be an alternative to treatment

Submitted by GABRIELA RONCADA HADDAD null (gabriela.haddad@yahoo.com) on 2017-09-10T22:08:03Z No. of bitstreams: 1 Tese Doutorado 27 jul 2017 - Gabriela.pdf: 1172239 bytes, checksum: d6ae9b8e2d02ba5890bf385b02539433 (MD5) / Approved for entry into archive by Monique Sasaki (sayumi_sasaki@hotmail.com) on 2017-09-12T16:46:39Z (GMT) No. of bitstreams: 1 haddad_gr_dr_bot.pdf: 1172239 bytes, checksum: d6ae9b8e2d02ba5890bf385b02539433 (MD5) / Made available in DSpace on 2017-09-12T16:46:39Z (GMT). No. of bitstreams: 1 haddad_gr_dr_bot.pdf: 1172239 bytes, checksum: d6ae9b8e2d02ba5890bf385b02539433 (MD5) Previous issue date: 2017-08-21 / Introdução: A influência do ácido retinoico (AR) sobre o coração é bastante relevante, ocorrendo durante a embriogênese, diferenciação e desenvolvimento cardíaco. Estudos experimentais mostram que o AR pode induzir hipertrofia excêntrica com melhora da função cardíaca em coração de ratos sadios, e também reduzir a hipertrofia, modulando o sistema renina angiotensina aldosterona (SRAA), em animais hipertensos. Pouco se sabe sobre os efeitos do AR em coração de humanos. Pacientes portadores de acne fazem uso de um tipo de AR que é o 13- cis-AR, também chamado de isotretinoína e por isso possibilitam o estudo do papel do AR em humanos. Estudo prévio mostrou que com 2 meses de uso do 13-cis-AR houve remodelação cardíaca. Entretanto, não se sabe sobre os mecanismos ou se essas alterações são reversíveis. Objetivos: Portanto, os objetivos desse trabalho foram de comparar a avaliação morfofuncional cardíaca e variáveis do SRAA entre pacientes em uso de isotretinoína com um grupo controle. Adicionalmente, avaliar se as alterações são reversíveis em pacientes em uso de isotretinoína. Casuística e Métodos: Foram estudados 35 adolescentes e adultos jovens, com idade entre 14 e 23 anos, do sexo masculino, sendo 20 deles em uso de isotretinoína oral, na dose de 0,5 mg/kg/dia a 0,75 mg/kg/dia, acompanhados no ambulatório de dermatologia do Hospital das Clínicas da Faculdade de Medicina de Botucatu (FMBUNESP), aos 6 meses de tratamento. Os outros 15 pacientes foram convidados na comunidade ou apresentavam acne leve com indicação apenas de tratamento tópico. Foram realizados avaliação morfofuncional e doppler tissular por meio de ecocardiografia transtorácica, dosagens bioquímicas de rotina e dosagens de componentes do SRAA renina, angiotensina I, angiotensina II, aldosterona, angiotensina 1-7 e alamandina. Essas variáveis foram comparadas nos grupos controle e isotretinoína pelo teste t de student ou Mann Whitnney. Os pacientes que receberam isotretinoína foram estudados antes do início do tratamento, com 6 meses de tratamento e 2 meses após o término do tratamento. Esses momentos foram comparados por meio do teste de Anova de 1 via de medidas repetidas. O nível de significância adotado foi de 5%. Resultados: Os pacientes do grupo controle e isotretinoína apresentaram a mesma idade, índice de massa corcoral, pressão arterial e frequência cardíaca. Dosagens bioquímicas habitualmente solicitadas durante o tratamento como enzimas hepáticas, função renal e triglicérides também foram semelhantes entre os grupos. Os dados morfológicos mostraram aumento do diâmetro diastólico do ventrículo esquerdo (DDVE) acompanhado de aumento do débito cardíaco e do fluxo transmitral avaliado por E/E’. Houve aumento do volume do átrio esquerdo (AE), no limite da significância e tendência ao aumento da massa do ventrículo esquerdo (VE) e com espessura relativa da parede semelhante entre os grupos. Sobre o SRAA houve redução da angiotensina II e da renina. Na avaliação ecocardiográfica apenas dos pacientes em uso de isotretinoína observou-se que houve redução do AE e do índice de massa do VE (IMVE) após 2 meses do término do tratamento. Embora não significativo, o comportamento de E/E’ também foi de redução após o tratamento. Discussão: O 13-cis-AR promove remodelação cardíaca, provavelmente induzida por hipervolemia, levando a um padrão de hipertrofia excêntrica com melhora da função. Essas alterações provavelmente levaram a menor ativação do SRAA visto pela redução da renina e angiotensina II. Esse perfil de remodelação e de bloqueio do SRAA é semelhante ao que ocorre no exercício físico. Nesse estudo foi possível apenas avaliar o grupo isotretinoína, quanto às variáveis ecocardiográficas antes, durante e após o tratamento. Observa-se que o término do estimulo com 13-cis-AR reduz algumas variáveis como átrio esquerdo e massa do VE. Portanto, em corações normais de adultos jovens, o AR atenuou o efeito de SRAA e promoveu remodelação cardíaca do tipo excêntrica, com melhora da função, compatível com sobrecarga volêmica e com caráter transitório. / Background: The influence of retinoic acid (RA) in the heart is very relevant, occurring during the embryogenesis, differentiation and cardiac development. Experimental studies shown that RA induces excentric hypertrophy, with improvement of cardiac function in heart of healthy rats. In addition, it was observed that RA regulates renin angiotensin aldosterone system (RAAS), a regulatory system involved in blood pressure, volume homeostasis and cardiac hypertrophy. There is a lack of information about the role of RA on cardiac remodeling in adult humans. Otherwise there are patients with Acne that uses 13- cis-RA, also called isotretinoin, and this population allow the investigation of cardiac remodeling and RA treatment. In fact, previously study shown that the use of 13-cis-RA, for acne, for 2 months induced cardiac remodeling, however, no one knows if these changes are persistent and reversible. Objectives: the objectives of these study is to compare the cardiac morphofunctional evaluation and RASS variables in patients using isotretinoin and in control group. Additionally, evaluate if these changes are reversible in isotretinoin group. Casuistic and Methods: Study1: 35 young men, between 14 to 23 years, 20 in use of 13-cis-RA, in the dose of 0,5 mg/kg/day to 0,75 mg/kg/day, from dermatology clinic of São Paulo State University (FMB-UNESP), at 6 months of treatment. The others 15 patients had mild acne only with topic treatment. Morphofunctional evaluation, tissular Doppler, Biochemical evaluation, dosage of RAAS components were performed. Results were compared in isotretinoin and control group by t student or Mann Whitnney tests. Study 2: Only the isotretinoin group were evaluated before beginning of treatment (initial moment - M0), after 6 months of treatment (final moment - M1) and two months after finishing the medication (M2). This results were compared by One way pared Anova. The statistically significant level was set at 5%. Results: control and isotretinoin group presents similar age, body mass index, blood pressure and heart rate. Biochemical evaluation was also similar. The present study showed that young patients receiving 13-cis-RA for Acne treatment presented left ventricle and atrium chamber enlargement and increase in cardiac output and in mitral flows. There was a trend toward higher ventricular mass with preserved relative wall thickness. RAAS showed decreased in angiotensin II and renin. Considering only patients that received isotretinoin, it was observed that cardiac atrium size and flows returned to baseline 2 months after the end of treatment and cardiac structures such as ventricle mass and thickness reduced. Discussion: 13-cis-RA promotes cardiac remodeling, probably induced by hypervolemia, taking to a pattern of eccentric hypertrophy, with improvement of function. It is possible that hypervolemia or a direct effect of 13-cis-RA, reduces renin and angiotensin II. The remodeling phenotype described is compatible with cardiac remodeling induced by physical activity, marked by hypervolemia, excentric hypertrophy and increased cardiac output. In the isotretinoin group, the end of treatment reduces left atrium size and left ventricle mass. Therefore, in normal hearts of young adults, RA reduces the effect of RAAS and promotes eccentric cardiac remodeling, with improvement of function, compatible with volume overload and with transitory character.

Orientador: Francisco Eduardo Martinez / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-18T07:43:37Z (GMT). No. of bitstreams: 1 Fontanelli_BeatrizAparecidaFioruci_M.pdf: 2160007 bytes, checksum: 5e358e9b41de70f7fd7c801fae0e0378 (MD5) Previous issue date: 2011 / Resumo: A exposição ao etanol altera a concentração do retinol e do all-trans-ácido retinóico (atAR) em vários tecidos. Os retinóides, retinol e atAR, são importantes para a diferenciação e manutenção das células epiteliais da próstata. O atAR se liga aos receptores de ácido retinóico (RARa, ß e y) e a interação receptor/ligante com a sequência responsiva ao retinóide no DNA, levam à transcrição de genes alvos. Assim, o atAR exerce efeitos no crescimento celular, diferenciação e apoptose, sendo essencial no desenvolvimento e diferenciação de órgãos e tecidos. Nosso objetivo foi analisar o retinol, o ácido retinóico e seus receptores, bem como, o índice de proliferação celular e de apoptose no lobo dorsolateral da próstata de ratos adultos UCh. Os animais foram divididos em quatro grupos experimentais (n=10/grupo): UChA (ingestão voluntária de etanol a 10% (v/v); UChACo (controle - ausência de etanol); UChB (ingestão voluntária de etanol a 10% (v/v) e UChBCo (controle - ausência de etanol). Após 150 dias de experimentação, os animais foram eutanasiados por decapitação e o sangue do tronco e os lobos dorsolaterais das próstatas foram coletados e processados: (1) para análises da concentração do retinol e do atAR no plasma e na próstata por meio de HPLC; (2) e análises de microscopia de luz para a proliferação celular (Ki-67), apoptose (Tunel) e para os receptores de ácido retinóico, por meio dos anticorpos anti-RARa, -ß e -y. O consumo crônico de etanol diminuiu a concentração do retinol no plasma dos grupos UChB (consumo alto de etanol) e UChA (consumo baixo de etanol). A concentração do retinol foi ainda menor no plasma do grupo UChB comparado ao UChA. No entanto, a concentração do retinol no tecido prostático não teve diferença significativa entre os grupos. O atAR aumentou significativamente somente no plasma do grupo UChB. Na próstata, a concentração do atAR aumentou no grupo UChB, enquanto que no UChA não houve diferença estatística. O RAR? na próstata dorsal e lateral dos ratos UCh não foi alterada em função do consumo de etanol. Já os RARß e -? apresentaram aumento do sinal na próstata dorsal do grupo UChB. Não houve diferença no índice de proliferação celular e de apoptose nas próstatas dorsais e laterais dos grupos experimentais. Conclui-se que o etanol altera a concentração do retinol e do atAR no plasma. Essa alteração é diretamente proporcional à quantidade de etanol consumida. Já na próstata, o retinol não é alterado pelo etanol. O consumo alto de etanol altera a concentração do atAR na próstata dorsolateral e a expressão dos RAR ß e y na próstata dorsal. A alteração da expressão dos RAR pode aumentar a sensibilidade da próstata à ação do atAR. O etanol não altera a proliferação celular e a apoptose na próstata dorsal e lateral / Abstract: Ethanol exposure alters the concentration of retinol and all-trans retinoic acid (atAR) in several tissues. Retinoids (retinol and atAR) are essential for the differentiation and homeostasis of the prostate epithelial cells. atAR binds to retinoic acid receptors (RAR a, ß and ?) and the interaction receptor/ligand with the sequence responsive to retinoid into DNA lead to transactivation of target genes. Thus, atAR directly produces their effects on cell growth, differentiation and apoptosis. This study aimed to analyze the retinol and all-trans-retinoic acid concentrations and its atAR receptors as well as the cell proliferation and apoptosis index upon the dorsolateral prostate lobe of adult UCh rats. All animals were divided into four experimental groups (n = 10/group): UChA (10% ethanol (v / v) voluntary intake); UChACo (without ethanol consumption); UChB (10% ethanol (v / v) voluntary intake) and UChBCo (without ethanol consumption). After 150 days of experimentation, animals were sacrificed followed by decapitation and trunk blood and dorsolateral prostate lobes collected. Samples of plasma and prostate by concentration analysis of the retinol and atAR were processed for HPLC. The cell proliferation and apoptosis immunoreactivities were assessed by Ki-67 and Tunel, respectively, and nuclear receptors by anti-RAR a,-ß and-y. Chronic ethanol consumption reduced the concentration of plasma retinol in UChB (high ethanol intake) and UChA groups (low ethanol intake). The retinol concentration in plasma was even lower in UChB compared to UChA group. However, the retinol concentration in prostate tissue was not significantly different between the groups. Concentration of atAR increased in plasma of UChB group, and was 96% higher in the UChA group. The prostate, atAR increased in the UChB group, while in UChA group no statistical difference. There was no statistical difference in proliferation cell and apoptosis in the dorsal and lateral prostate lobes between the groups. The expression of RAR a in the dorsal and lateral prostate of UCh rats was not altered as a function of ethanol consumption. Already RAR ß and-y showed increased signal in the dorsal prostate UChB group. We conclude that ethanol alters the concentration of retinol and atAR in plasma. This change is directly proportional to the amount of ethanol consumed. In the prostate, retinol is not altered by ethanol. The high ethanol intake alters the concentration of atAR in dorsolateral prostate and the expression of RARß and RARy in the dorsal prostate. Alteration in expression of RAR can increase sensitivity to the action of the atAR in prostate. Ethanol does not alter cell proliferation and apoptosis in the dorsolateral prostate / Mestrado / Anatomia / Mestre em Biologia Celular e Estrutural

Os retinóides têm sido amplamente utilizados na clínica dermatológica e nos produtos cosméticos com finalidades preventivas e reparadoras dos efeitos indesejáveis do envelhecimento cutâneo. Considerando que a concentração de retinóides, como por exemplo, o ácido retinóico ou o palmitato de retinila, pode influenciar na eficácia e segurança de uso dos mesmos em formulações tópicas, é de fundamental importância o desenvolvimento e avaliação dos efeitos benéficos e/ou colaterais de formulações dermocosméticas contendo diferentes concentrações de ácido retinóico (0,025%, 0,05% e 0,1%) e palmitato de retinila (0,25%, 0,5% e 1,0%), visando a obtenção de uma concentração que proporcione a máxima eficácia possível e risco mínimo à pele. Assim, o presente trabalho tem por objetivos avaliar a influência de diferentes concentrações de retinóides (ácido retinóico ou palmitato de retinila) em formulações dermocosméticas com finalidades antienvelhecimento, na pele de camundongos sem pêlo, por estudos histopatológicos, morfométricos, estereológicos e por Bioengenharia Cutânea. Para tal foram preparadas três formulações de géis creme à base de hidroxietilcelulose (HEC) e microemulsão de silicone e octanoato de octila (formulações de nos 1 e 3) e à base de complexo lipídico contendo álcool batílico e lecitina de soja, HEC e octanoato de octila (formulação de nº 2) as quais foram submetidas a testes preliminares de estabilidade. A formulação de nº 1 (F1) foi considerada a mais estável e, portanto, selecionada como veículo para a avaliação dos efeitos do ácido retinóico e do palmitato de retinila na pele de camundongos sem pêlo. Para a realização do ensaio biológico, amostras das formulações, acrescidas ou não (veículo) de 0,025; 0,05 ou 0,1% ácido retinóico ou 0,25; 0,5 ou 1,0% de palmitato de retinila foram aplicadas no dorso de camundongos sem pêlo. Após cinco dias da aplicação diária destas formulações, foram obtidas medidas de índice de eritema pelo equipamento Mexameter® MX16 e medidas do conteúdo aquoso do estrato córneo pelo equipamento Corneometer® CM825. Em seguida, os camundongos foram mortos e posteriormente foram colhidos fragmentos de pele das áreas que receberam aplicação das formulações, bem como da área que não foi aplicada nenhuma formulação (controle) e, a seguir, obtidos cortes histológicos para a realização dos estudos histopatológicos, morfométricos e estereológicos. De acordo com as metodologias empregadas, foi possível observar que, na avaliação do conteúdo aquoso do estrato córneo, somente as formulações que continham ácido retinóico em diferentes concentrações, provocaram mudanças significativas, reduzindo este conteúdo, ou seja, ocasionou um ressecamento na superfície da pele. Apenas as formulações contendo 0,025 e 0,1% de ácido retinóico e 1,0% de palmitato de retinila provocaram um aumento no índice de eritema. Além disso, tanto o ácido retinóico quanto o palmitato de retinila, atuaram na epiderme, porém de modo e intensidade diferentes, sendo que, o ácido retinóico teve um efeito mais pronunciado em relação às variáveis estudadas. As três diferentes concentrações de ácido retinóico e de palmitato de retinila ocasionaram aumento significante da espessura das camadas epiteliais, sem alteração da camada córnea. O ácido retinóico e o palmitato de retinila atuaram ainda aumentando os volumes nuclear, citoplasmático e celular, um dos fatores que ocasionou aumento da espessura do epitélio. Finalizando, ao analisar todas as variáveis histopatológicas, morfométricas e estereológicas, bem como o índice de eritema e o conteúdo aquoso do estrato córneo estudados, podemos sugerir que as formulações que continham as concentrações intermediárias, tanto do ácido retinóico (0,05%) quanto do palmitato de retinila (0,5 %), foram as que apresentaram melhores resultados, principalmente no que se refere a uma relação risco / benefício adequada e aceitável. / Retinoids has been widely used in dermatological clinic and in cosmetics products with preventive purposes as well as for the repairmen of the cutaneous aging undesirable effects. Considering that the concentration of retinoids, i.e., the retinoic acid or the retinyl palmitate, can influence their efficacy and safety in topical formulations, the development and evaluation of the beneficial and/or collateral effect of dermocosmetic formulations containing different concentrations of retinoic acid (0.025%, 0.05% e 0.1%) and retinyl palmitate (0.25%, 0.5% e 1.0%) is very important, aiming at the attainment of a concentration that provides to the maximum possible efficacy and minimum risk to the skin. Thus, the aim of this study was to evaluate the influence of different concentrations of retinoids (retinoic acid or retinyl palmitate) in dermocosmetics formulations with antiaging purposes, in hairless mice, using histopathological, morphometric and stereologic studies and Skin Bioengineering Techniques. For this purpose, three gel cream formulations were developed, containing hydroxyethylcellulose (HEC) and silicone microemulsion and octyl octanoate (formulations nº 1 and 3) and a complex lipidic based formulation containing batyl alcohol and lecithin, HEC and octyl octanoate (formulation nº 2), which were submitted to preliminary stability tests. The formulation nº1 (F1) was considered the most stable, therefore, it was selected as the vehicle for the evaluation of the effects of the retinoic acid and the retinyl palmitate in hairless mice skin. For the accomplishment of the biological assay, samples of the formulations, supplemented or not (vehicle) of 0.025, 0.05 or 0.1% retinoic acid or 0.25, 0.5 or 1.0% retinyl palmitate were applied in the dorsal skin of hairless mice. After five days of daily application of these formulations, the erythema index was measured by reflectance spectrophotometry using a Mexameter® MX16 as well as the water content of the stratum corneum using Corneometer® CM825. After that, the hairless mice were sacrificed and later skin fragments were obtained for each area that received application of the formulations, as well as of the area that was not applied any formulation (control) and, after that histologic sections were obtained and submitted to histopathological, morphometric and stereologic studies. In accordance with the used methodologies, it was possible to observe that in the evaluation of the water content of the stratum corneum, only the formulations that contained retinoic acid in different concentrations provided significant changes, enhancing skin surface dryness. Only the formulations containing 0.025 and 0.1% of retinoic acid and 1.0% of retinyl palmitate provided an increase in erythema index. Moreover, both retinoic acid and retinyl palmitate acted in the epidermis, however in different intensity and way, since retinoic acid had more pronounced effects in relation to the studied variables. The three different concentrations of retinoic acid and retinyl palmitate caused a significant increase of the epithelial layers thickness, without alteration of the horny layer. Retinoic acid and retinyl palmitate also increased the nucleus, cytoplasmic and cell volumes, which was one of the factors that influenced the increase of the epithelium thickness. Finishing, when analyzing all the histopathological, morphometric and stereologic variables, as well as the erythema index and the water content of the stratum corneum studied, we can suggest that the formulations that contained the intermediate concentrations (such as 0.05% of retinoic acid and 0.5% of retinyl palmitate) presented the best results, mainly when an adequate and acceptable risk/ benefit relationship is considered.