Ce travail de thèse avait pour objectif d’élaborer un système vecteur ciblantl’endothélium pathologiquement activé dans les tissus enflammés, infectés ou tumoraux.Ce vecteur est sous forme de nanoparticules décorées de ligands glycosides capablesd’interagir avec l’E-sélectine, un récepteur exprimé sur les cellules endothélialesactivées.Nous avons mis au point une synthèse de copolymère amphiphile avec une architectureà bloc muni sur sa partie hydrophile d’un ligand glucidique. Ce copolymère a été par lasuite utilisé pour la préparation de nanoparticules de type coeur/couronne. La partiehydrophobe centrale est entourée d’une couronne hydrophile dont l'encombrementstérique et la mobilité limitent l'opsonisation de la particule. Le ciblage actif a été assurépar la présence d’un ligand du récepteur de l’E-sélectine aux extrémités des chaînes depolymères hydrophiles à la surface du vecteur.Avec ces nanoparticules dont les propriétés de surface sont prédéfinies, nous avonsmontré in vitro l’association efficace avec les cellules endothéliales activées, ce qui apermis de valider ce concept de ciblage moléculaire actif par l’intermédiaire du couplerécepteur/ ligand. Un tel système permettra d’améliorer l’indice thérapeutique et labiodistribution des principes actifs anti-inflammatoire et/ou anticancéreux. / The objective of this work was to develop a delivery system targeting pathologically activated endothelium within inflamed, infectious, and some tumoral tissues. This system is composed of nanoparticles bearing sugar residues that are able to recognize and interact with E-selectin, a receptor expressed on the activated endothelial cells.We synthesized an amphiphilic block copolymer with the hydrophilic part terminated by a carbohydrate ligand. The construction was achieved by a combination of click chemisty, ring-opening polymerization and atom transfer radical polymerization. This copolymer was used to prepare nanoparticles of the core/shell type where the central hydrophobic body is surrounded with the hydrophilic shell that can stabilize the particles in aqueous media and limit their opsonisation. Active targeting was achieved by coupling an analogue of sialyl Lewis X, the physiological ligand of E-selectin to the end of the hydrophilic polymer chains on the surface of the particles.We were able to demonstrate in vitro the efficient association of these nanoparticles with defined surface properties with activated endothelial cells. This allowed us to validate our concept of active molecular targeting using this couple receptor/ligand couple. Such a system could be used to improve the therapeutically index and the biodistribution of anti-inflammatory and anti-tumor drugs.

INTRODUÇÃO: A Artrite Reumatóide (AR) é caracterizada pelo aspecto inflamatório crônico articular e é a doença autoimune mais comum em todo o mundo. A categoria de medicamentos modificadores do curso da doença (MMCD) é dividido em dois grupos, sintéticos e biológicos. Há controversos estudos em relação a comparação entre estas alternativas, principalmente, devido ao elevado custo dos biológicos. O objetivo deste estudo foi realizar uma avaliação econômica do tratamento da AR, comparando a terapia combinada de MMCD sintéticos versus MMCD biológicos, utilizando de dados da literatura e de custo na perspectiva do SUS. MÉTODOS: Foi realizado uma revisão sistemática com metanálise das bases Medline e Embase os ensaios clínicos randomizados(ECR) que fizeram comparação direta entre o uso de MMCD sintéticos versus os MMCD biológicos. A remissão foi considerada para DAS28 < 2,6. Os itens que compõe o custo seguiram a diretrizes do Ministério da Saúde do Brasil e os valores foram recuperados da tabela do Sistema de Informação Ambulatorial do SUS, e do Sistema de Gerenciamento da Tabela de Procedimentos, Medicamentos e OPM do SUS, dados do ano 2016/2017. RESULTADOS: Foram incluídos 6 ECR. No seguimento de até 6 meses o RR=0,70 (IC95% 0,57 a 0,85) a favor dos biológicos. No seguimento entre 12 a 24 meses não houve diferença estatisticamente significante, RR=0,91 (IC95% 0,80 a 1,05). Um ano do tratamento com MMCDs combinado custa R$2445,60 e os Anti-TNF custa R$ 52.821,57. CONCLUSÃO: A remissão da atividade clínica da AR pode ser obtida pelo uso de DMARD sintéticos ou por Agentes Biológicos. Análise de custo-minimização mostrou que uma economia substancial a cada mês evitado de uso dos MMCD biológicos / INTRODUCTION: Rheumatoid arthritis (RA) is the most common autoimmune disease in the world, which leads to a chronic joint inflammation. There are two types of disease-modifying anti-rheumatic drugs (DMARD): synthetical and biological. The comparison between both drugs is controversial, mostly because of the high cost of the biological ones. The aim of this study was to develop an economic evaluation of RA treatments, comparing combined therapy with synthetic DMARD versus biological DMARD, based on literature review and cost analysis on SUS data. METHODS: Systematic review with meta-analysis of randomized clinical trials (RCT) was conducted on Medline and Embase database about direct comparisons of synthetic DMARD and biological DMARD. Remission was set for DAS28 < 2.6. Cost analysis was based on the guidelines of the Brazilian Ministry of Health and cost values were extracted from the SUS\'s Ambulatory Information System table, Management System of Procedures Table, and Medicine and OPM table, for 2016/2017. RESULTS: Six RCT were included. For six months follow-up, RR=0.70 (IC95% 0.57 to 0.85) in favour of biologicals. For 12-24 months follow-up, both DMARD were similar, RR=0.91 (IC95% 0.80 to 1.05). One-year treatment with DMARD costs R$2445,60 e Anti-TNF costs R$ 52.821,57. CONCLUSION: Remission of clinical activity of RA can be reached with synthetic or biologic DMARD. Minimizing-cost analysis showed a monthly expressive saving avoinding the biologic DMARD

Objetivo: avaliar marcadores de reserva ovariana e a presença de anticorpo anti-corpo lúteo (anti-CoL) em pacientes com lúpus eritematoso sistêmico (LES) de início na infância. Métodos: A presença do anti-CoL foi avaliada através de immunoblot em cinquenta e sete mulheres com LES e 21 controles saudáveis. A reserva ovariana foi estimada através das dosagens do hormônio folículo estimulante (FSH), hormônio luteinizante (LH), estradiol, hormônio anti- Mülleriano (AMH) e da contagem de folículos antrais (CFA). Foram também avaliados dados demográficos, alterações menstruais, atividade da doença, dano cumulativo e tratamento. Resultados: a mediana da idade atual foi similar nos pacientes com LES em relação aos controles (27,7 vs. 27,7 anos, p=0,414). A mediana do AMH (1,1 vs. 1,5ng/mL, p=0,037) da CFA (6 vs. 16 p<0,001) forma significantemente menores nos pacientes com LES quando comparados aos controles, porém sem alterações menstruais significantes. A presença do anti-CoL foi observada apenas nos pacientes com LES (16% vs. 0%, p=0,103) e não foi relacionada com dados demográficos, parâmetros de reserva ovariana, atividade da doença, dano cumulativo ou tratamento. Avaliação dos pacientes tratados com ciclofosfamida mostrou níveis elevados de FSH quando comparados com os pacientes que não receberam ciclofosfamida e com controles (8,8 vs. 5,7 vs. 5,6IU/L, p=0,032) e níveis menores de AMH e CFA (0,4 vs. 1,5 vs. 1,5ng/mL, p=0,004; 4,0 vs. 6,5 vs. 16IU/L, p=0,001; respectivamente). Dezenove pacientes foram tratados com metotrexate sem histórico de uso de ciclofosfamida sendo evidenciada uma correlação negativa entre a dose cumulativa de metotrexate e os níveis de AMH (p=0,027, r=-0,507). Conclusões: este estudo identificou que altas doses ciclofosfamida e metotrexato são causas relevantes de disfunção ovariana subclínica durante a idade reprodutiva em mulheres com LES de início na infância / Objective: To assess ovarian reserve markers and anti-corpus luteum antibodies (anti-CoL) in systemic lupus erythematosus (SLE) patients with onset before adulthood. Methods: Fifty-seven SLE female patients and 21 healthy controls were evaluated for anti-CoL by immunoblot. Ovarian reserve was assessed by: follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol, anti-Müllerian hormone (AMH) and antral follicle count (AFC). Demographic data, menstrual abnormalities, disease activity, damage and treatment were also analyzed. Results: The median of current age was similar in SLE patients and controls (27.7. vs. 27.7 years, p=0.414). The median of AMH (1.1 vs. 1.5ng/mL, p=0.037) and AFC (6 vs. 16, p<0.001) were significantly reduced in SLE patients versus controls without significant menstrual abnormalities. Anti-CoL was solely observed in SLE patients (16% vs. 0%, p=0.103) and not associated with demographic data, ovarian reserve parameters, disease activity/damage and treatment. Further evaluation of patients treated with cyclophosphamide revealed a higher median of FSH levels compared to SLE patients not treated with cyclophosphamide and controls (8.8 vs. 5.7 vs. 5.6IU/L, p=0.032) and a lower median AMH levels and AFC (0.4 vs. 1.5 vs. 1.5ng/mL, p=0.004; 4.0 vs. 6.5 vs. 16IU/L, p=0.001; respectively). Nineteen patients were treated with methotrexate without cyclophosphamide use, and a negative correlation was observed between cumulative methotrexate dose and AMH levels (p=0.027, r=-0.507). Conclusions: The present study identifies high doses of cyclophosphamide and methotrexate as relevant causes of subclinical ovarian dysfunction during reproductive ages in SLE patients with onset before adulthood

Objetivo: O presente estudo teve como objetivo verificar os efeitos do tratamento com altas e baixas doses de metotrexato (MTX) na reparação de fraturas mandibulares, num modelo experimental com ratos. Métodos: O modelo experimental empregado consiste na criação de um defeito ósseo na mandíbula do rato, semelhante a uma fratura. Oitenta ratos foram distribuídos em 4 grupos de 20 animais que receberam, por via intraperitoneal: soro fisiológico (controle) (1 ml, após a cirurgia); dexametasona (DX) (0,15 mg / kg, dose única após a cirurgia); alta dose MTX (1,6 mg / kg, semanalmente); baixa dose MTX, (0,25 mg / kg, semanalmente). Os animais foram sacrificados no dia 1º, 7º, 15º e 30º dia após a cirurgia. As mandíbulas foram submetidas à análise radiográfica para medir a distância entre os cotos ósseos e a área de osteotomia. Avaliação histomorfométrica foi realizada usando um software analisador de imagens digitalizadas para verificar a formação de cartilagem e óssea. Resultados: Os resultados revelaram não haver alterações significantes entre os tratamentos nos parâmetros avaliados nos 10 e 70 dias após a cirurgia. Animais do grupo controle sacrificados no 15º dia após a cirurgia tiveram uma redução da distância entre as extremidades ósseas e na área da osteotomia, bem como um grande aumento na formação de cartilagem. O padrão desses parâmetros nos animais tratados com baixas doses de MTX e DX não foram significativamente diferentes do grupo controle neste período. No entanto, os animais tratados com alta dose de MTX tiveram aumento da distância entre os cotos ósseos e da área da osteotomia, bem como foi praticamente nulo o aumento da formação de cartilagem. Sobre o 30º dia após a cirurgia, os animais do grupo controle tinham praticamente recuperado a região da fratura, bem como aqueles tratados com doses baixas de MTX. Os grupos tratados com a alta dose de MTX e DX mantiveram abertos os defeitos ósseos. Conclusões: Este estudo mostra claramente que a baixa dose de MTX não afetou a reparação óssea de fraturas mandibulares em ratos, em contraste com a alta dose de MTX, que afeta desfavoravelmente a regeneração óssea. / Purpose: The present study aims to verify the effect of high and low dose of methotrexate (MTX) treatment on bone repair of mandibular fractures in rats. Methods: The experimental model employed consists in creating a defect in rat mandible, similar to a fracture. Eighty rats were distributed in 4 groups of 20 animals that received, intraperitoneally: saline (1 ml, after surgery); dexametazone (DX, 0.15 mg/Kg, one dose at surgery); high dose MTX (1.6 mg/Kg, weekly); low dose MTX, (0.25 mg/Kg, weekly). Groups of five animals were sacrificed on the 1st, 7th, 15th and 30th day after surgery. Mandibles were submitted to radiographic analysis to measure the distance between bony edges and the area of osteotomy. Histopathological evaluation was performed in digitalized images using an analyzer software to examine cartilage and bone formation. Results: The treatments did not alter any evaluated parameters on days 1 and 7 after surgery. Control animals sacrificed on the 15th day after surgery had a reduction of the distance between bony ends and in the area of osteotomy, as well as a great increase in cartilage formation. The pattern of these parameters in animals treated with low dose MTX and DX did not differ from control group. However, animals treated with higher dose of MTX kept increasing the distance between bony ends and the area of osteotomy, and the increase in cartilage formation was practically inexistent. On the 30th day after surgery, control animals had pratically recovered the fracture region, as well as those treated with low dose MTX. The group treated with higher dose MTX and DX still had an open bone defect. Conclusions: This study clearly shows that low dose MTX did not affect bone healing of mandibular fractures in rats, in contrast to a higher dose, which promotes impairment of bone regeneration.

Orientador: Maria Virgínia Costa Scarpa / Coorientador: Marlus Chorilli / Banca: Renata Fonsenca Vianna Lopez / Banca: Leila Aparecida Chiavacci / Resumo: Amplamente utilizado no tratamento de vários tipos de câncer e na psoríase, o metotrexato (MTX) é um quimioterápico, estruturalmente análogo do ácido fólico, que apesar de sua eficácia apresenta uma série de efeitos adversos, sendo a hepatotoxicidade o mais grave. Atualmente os sistemas nanoestruturados líquido-cristalinos de fase lamelar estão sendo utilizados como dispositivos para liberação modificada de fármacos, sendo vantajosos na liberação tópica de várias substâncias, conforme suas características de interação com o estrato córneo e as outras camadas da pele, evitando assim efeitos adversos sistêmicos. Os objetivos deste trabalho foram desenvolver sistemas nanoestruturados líquido-cristalinos de fase lamelar, acrescidos de MTX, caracteriza-los do ponto de vista físico, realizar a análise estrutural das formulações, através de microscopia de luz polarizada (MLP), espalhamento de raios-X a baixo ângulo (SAXS) e suas propriedades reológicas, executar os testes de estabilidade preliminar (TEP) das formulações, validar o método analítico para a quantificação de MTX por CLAE e executar ensaios de liberação, permeação e retenção in vitro. As formulações preparadas a partir da mistura do poliéter funcional siloxano (Dow Corning® 5329) como tensoativo, com silicone fluido de co-polímero glicol (Dow Corning® 193C) como fase oleosa titulados em fase aquosa, composta por tampão fosfato de potássio monobásico 0,01M pH 7,4, apresentaram fases líquido-cristalinas do tipo lamelar, confirmados pelos ensaios de MLP e SAXS. Os TEPs evidenciaram que as formulações A, B e C mantiveram-se estáveis durante o período do estudo. Os estudos do comportamento reológico das formulações apresentaram-se como fluidos pseudoplásticos não-newtonianos tixotrópicos ... / Abstract: Widely used in the treatment of some types of cancer, the methotrexate (MTX) is a chemotherapeutic, structurally analog of the folic acid, although its effectiveness, presents a series of adverse effect, being the most serious hepatotoxicity. Currently, the liquid crystal lamellar phase is being used of devices for modified release of drug demonstrated to be advantageous in the release topic of some substances, given to the characteristics of interaction with the stratum corneun and other layers of the skin, avoiding systemic adverse effects. The aims of this research had been to develop and to characterize liquid crystalline nanostructure systems of lamellar phase, increased of MTX of the physical point of view, also carry through the structural analysis of the formulations through by polarized light microscopy (PLM), small-angle X-ray scattering (SAXS) and rheological properties. Perform stability studies of the chosen formulations, validate the analytical method of quantification of MTX for High Performance Liquid Chromatography (HPLC) and carry through release assay, cutaneous permeation and skin retention in vitro for the chosen formulations. The formulations prepared by the mixture of polyether functional siloxane as surfactant, with silicone polyether copolymer as oily phase and phosphate buffer 0,01M pH 7,4 as aqueous phase demonstrating lamellar liquid-crystalline phases, confirmed by assays of PLM and SAXS. The stability studies showed that formulations A, B and C remained stable throughout the period of the study. The study of the rheological behavior of the formulations presented as not Newtonian pseudoplastic fluid and thixotropic ... / Mestre

Objetivo: avaliar marcadores de reserva ovariana e a presença de anticorpo anti-corpo lúteo (anti-CoL) em pacientes com lúpus eritematoso sistêmico (LES) de início na infância. Métodos: A presença do anti-CoL foi avaliada através de immunoblot em cinquenta e sete mulheres com LES e 21 controles saudáveis. A reserva ovariana foi estimada através das dosagens do hormônio folículo estimulante (FSH), hormônio luteinizante (LH), estradiol, hormônio anti- Mülleriano (AMH) e da contagem de folículos antrais (CFA). Foram também avaliados dados demográficos, alterações menstruais, atividade da doença, dano cumulativo e tratamento. Resultados: a mediana da idade atual foi similar nos pacientes com LES em relação aos controles (27,7 vs. 27,7 anos, p=0,414). A mediana do AMH (1,1 vs. 1,5ng/mL, p=0,037) da CFA (6 vs. 16 p<0,001) forma significantemente menores nos pacientes com LES quando comparados aos controles, porém sem alterações menstruais significantes. A presença do anti-CoL foi observada apenas nos pacientes com LES (16% vs. 0%, p=0,103) e não foi relacionada com dados demográficos, parâmetros de reserva ovariana, atividade da doença, dano cumulativo ou tratamento. Avaliação dos pacientes tratados com ciclofosfamida mostrou níveis elevados de FSH quando comparados com os pacientes que não receberam ciclofosfamida e com controles (8,8 vs. 5,7 vs. 5,6IU/L, p=0,032) e níveis menores de AMH e CFA (0,4 vs. 1,5 vs. 1,5ng/mL, p=0,004; 4,0 vs. 6,5 vs. 16IU/L, p=0,001; respectivamente). Dezenove pacientes foram tratados com metotrexate sem histórico de uso de ciclofosfamida sendo evidenciada uma correlação negativa entre a dose cumulativa de metotrexate e os níveis de AMH (p=0,027, r=-0,507). Conclusões: este estudo identificou que altas doses ciclofosfamida e metotrexato são causas relevantes de disfunção ovariana subclínica durante a idade reprodutiva em mulheres com LES de início na infância / Objective: To assess ovarian reserve markers and anti-corpus luteum antibodies (anti-CoL) in systemic lupus erythematosus (SLE) patients with onset before adulthood. Methods: Fifty-seven SLE female patients and 21 healthy controls were evaluated for anti-CoL by immunoblot. Ovarian reserve was assessed by: follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol, anti-Müllerian hormone (AMH) and antral follicle count (AFC). Demographic data, menstrual abnormalities, disease activity, damage and treatment were also analyzed. Results: The median of current age was similar in SLE patients and controls (27.7. vs. 27.7 years, p=0.414). The median of AMH (1.1 vs. 1.5ng/mL, p=0.037) and AFC (6 vs. 16, p<0.001) were significantly reduced in SLE patients versus controls without significant menstrual abnormalities. Anti-CoL was solely observed in SLE patients (16% vs. 0%, p=0.103) and not associated with demographic data, ovarian reserve parameters, disease activity/damage and treatment. Further evaluation of patients treated with cyclophosphamide revealed a higher median of FSH levels compared to SLE patients not treated with cyclophosphamide and controls (8.8 vs. 5.7 vs. 5.6IU/L, p=0.032) and a lower median AMH levels and AFC (0.4 vs. 1.5 vs. 1.5ng/mL, p=0.004; 4.0 vs. 6.5 vs. 16IU/L, p=0.001; respectively). Nineteen patients were treated with methotrexate without cyclophosphamide use, and a negative correlation was observed between cumulative methotrexate dose and AMH levels (p=0.027, r=-0.507). Conclusions: The present study identifies high doses of cyclophosphamide and methotrexate as relevant causes of subclinical ovarian dysfunction during reproductive ages in SLE patients with onset before adulthood

<p>Uvod: Reumatoidni artritis (RA) je bolest koja dovodi do ireverzibilnog o&scaron;tećenja zglobova usled čega je neophodno pri postavljanju dijagnoze započeti lečenje. TNF-alfa inhibitori predstavljaju revolucionarno otkriće u lečenju RA, pri čemu su najče&scaron;će kori&scaron;ćeni Etanercept i Adalimumab. Oni nisu efikasni kod svih pacijenata kod kojih se primene, a mehanizmi gubitka odgovora nisu jasni. Cilj rada je odrediti uticaj Etanercepta i Adalimumaba na aktivnost bolesti (merenjem DAS28 SE i DAS28 CRP skora) i funkcionalni status pacijenata (merenjem HAQ-DI upitnika), broj bolnih i otečnih zglobova pre i tokom godinu dana primene ovih lekova, kao i utvrditi povezanost koncentracije Etanercepta i Adalimumaba u krvi sa vrednostima DAS28 SE u momentu odreĎivanja koncentracije leka. Praćena je i učestalost neželjenih efekata kod pacijenata lečenih sa ova dva leka. Ispitan je i uticaj primene Metotreksata na nivoe lekova u krvi, kao i doza Metotreksata pre i 6 meseci nakon uvoĎenja Etanercepta ili Adalimumaba. Metode: Studija je sprovedena u Specijalnoj bolnici za reumatske bolesti i Klinici za nefrologiju i kliničku imunologiju, Kliničkog centra Vojvodine u Novom Sadu i obuhvatila je 88 pacijenata kod kojih je postavljena dijagnoza RA, od kojih je 49 bilo lečeno Etanerceptom, a 39 Adalimumabom. Analizirana je medicinska dokumentacija, a nakon početka primene TNF-alfa inhibitora svim ispitanicima je u toku godinu dana svaka tri meseca raĎena kontrola koja je podrazumevala anamnezu i fizički pregled, analizu biohemijskih nalaza krvi, merena je aktivnost bolesti merenjem indeksa aktivnosti bolesti DAS28 SE i DAS28 CRP i raĎena procena funkcionalnog statusa tako &scaron;to je pacijent popunjavao HAQ-DI upitnik. Rezultati: Aktivnost RA merena DAS28 SE i DAS28 CRP indeksima, funkcionalni status meren HAQ-DI upitnikom, broj bolnih i otečenih zglobova i vrednosti reaktanata akutne faze značajno su veći pre početka terapije Etanerceptom i Adalimumabom i smanjuje se tokom prvih 6 meseci lečenja ovim lekovima i potom se taj efekat terapije održava do kraja perioda praćenja. Nema statistički značajne razlike u poreĎenju Etanercepta i Adalimumaba u odnosu na učestalost neželjenih dejstava. Doza Metotreksata je statistički značajno manja 6 meseci nakon upotrebe biolo&scaron;kog leka Etanercept i Adalimumab. Pacijenti lečeni Metotreksatom uz Adalimumab imali su statistički značajno veće nivoe leka, nego oni koji ga nisu koristili. Zaključak: TNF-alfa inhibitori ne dovode do zaustavljanja bolesti kod svih pacijenata kod kojih se primene. Mehanizam gubitka odgovora na terapiju TNF-alfa inhibitorima nije jasan. Kako bi se donela najbolja odluka za pacijenta, neophodno je odrediti nivo leka u krvi, kao i nivo antitela na lek prilikom svake promene stanja pacijenta. Za sada nema dovoljno studija koje ukazuju da li postoji veza izmeĎu ekspresije TNF-alfa gena i nivoa TNF-alfa u krvi, te da li bi se merenjem TNF-alfa u krvi mogla korigovati terapija i doza TNF-alfa inhibitora &scaron;to će verovatno biti predmet budućih istraživanja.</p> / <p>Rheumatoid Arthritis (RA) is a disease that leads to irreversible joint damage, which makes necessary to start treatment when the diagnosis is set. TNF-alpha inhibitors represent a revolutionary discovery in the treatment of RA, and the most commonly used are Etanercept and Adalimumab. They are not effective in all patients, and the mechanisms of loss of response are not clear. The aim of this study is to determine the effect of Etanercept and Adalimumab on disease activity (by measuring DAS28 SE and DAS28 CRP score) and the functional status of patients (by measuring the HAQ-DI questionnaire), the number of painful and swollen joints before and during the first year of administration of these drugs. Also, it was determined a correlation between the concentration of Etanercept and Adalimumab in blood and the values of DAS28 SE at the moment of drug concentration measurement. The incidence of adverse effects in patients treated with these two drugs was also observed. It was examined the effect of Methotrexate on drug levels in the blood as well as the dose of Methotrexate before and 6 months after the introduction of Etanercept or Adalimumab. Methods: The study was conducted at the Special Hospital for Rheumatic Diseases and the Clinic of Nephrology and Clinical Immunology, Clinical Center of Vojvodina in Novi Sad. It included 88 patients with RA, 49 were treated with Etanercept and 39 with Adalimumab. Medical documentation was analyzed, and during the first year of TNF-alpha inhibitor administration, every three months were done anamnesis and physical examination, analysis of blood biochemical findings, measurements of the disease activity with DAS28 SE and DAS28 CRP score and a functional status assessment with the HAQ-DI questionnaire. Results: Disease activity measured by DAS28 SE and DAS28 CRP scores, functional status measured with HAQ-DI questionnaire, number of painful and swollen joints and acute phase reactant values are significantly higher before Etanercept and Adalimumab therapy and decreased during the first 6 months of treatment with these drugs and then this effect of therapy is maintained until the end of the monitoring period. There is no statistically significant difference in the comparison of Etanercept and Adalimumab with respect to the frequency of adverse events. The dose of Methotrexate was statistically significantly lower for 6 months after the use of Etanercept and Adalimumab. Patients treated with Methotrexate and Adalimumab had statistically significantly higher drug levels than those who did not use it. Conclusion: TNF-alpha inhibitors are not effective in all patients who used them. The mechanism of loss of response to TNF-alpha inhibitors is not clear. In order to make the best decision for the patient, it is necessary to determine the drug level in the blood as well as the level of antibodies to the drug in each change in the patient&#39;s condition. For now, there are not enough studies to indicate whether there is a link between expression of the TNF-alpha gene and the level of TNF-alpha in the blood, and whether the measurement of the TNF-alpha in blood could be used for therapy correction and change of dose of TNF-alpha inhibitor, which is likely to be the subject of the future research.</p>

No description available.

Le dihydrofolate réductase (DHFR) est la principale cible du méthotrexate, un important composant du traitement de la leucémie lymphoblastique aiguë (LLA). Une association des polymorphismes du promoteur de DHFR avec l’issue de la LLA a été mise en évidence au laboratoire. Une survie sans événement (EFS) réduite corrélait avec les allèles A -317 et C -1610, et l’haplotype *1, défini par ces allèles. L’haplotype *1 était aussi associé à une expression élevée du DHFR. Dans cette étude, nous étendons l’analyse à la région régulatrice adjacente, d’environ 400 pb, correspondant au transcrit mineur non-codant du DHFR, qui joue un rôle essentiel dans la régulation de la transcription au niveau du promoteur majeur. Six polymorphismes ont été identifiés, parmi lesquels 5 étaient des SNPs et un polymorphisme de longueur composé d’un nombre variable d’éléments de 9 pb et d’une insertion/délétion de 9 pb. L’analyse d’haplotype, incluant tous les polymorphismes promoteurs, a révélé une diversification de l’haploytpe *1 en 5 sous-types (*1a à *1e). Les variations du promoteur majeur et les sous-types de l’haplotype *1 ont été par la suite analysés pour l’association avec l’issue de LLA. Un EFS réduit corrélait avec l’allèle A du polymorphisme G308A (p=0,02) et avec l’haplotype *1 (p=0,01). Des niveaux élevées d’ARNm étaient trouvés chez les porteurs de l’haplotype *1b (p=0,005) et pas pour les autres sous-types de l’haplotype *1. Alors, la mauvaise issue de LLA associée avec l'haplotype *1 est en effet déterminée par le sous-type *1b. Cette étude donne un nouvel aperçu des polymorphismes régulateurs du DHFR définissant plus précisément les variations du DHFR prédisposant un événement. / Dihydrofolate reductase (DHFR) is the major target of methotrexate, a key component in childhood acute lymphoblastic leukemia (ALL) treatment. We recently reported an association of DHFR promoter polymorphisms with ALL outcome. Lower event free survival (EFS) correlated with the alleles A -317 and C -1610, and with haplotype *1, defined by these alleles. Haplotype*1 was also associated higher DHFR expression. Here we extended the analysis to adjacent 400bp regulatory region corresponding to non-coding minor DHFR transcript which plays an essential role in the regulation of transcription from the major promoter. Six polymorphisms were identified, of which 5 were SNPs and one length polymorphism composed of variable number of 9bp elements and 9bp insertion/deletion. Haplotype analysis including all promoter polymorphisms revealed diversification of haplotype *1 into 5 subtypes (*1a to *1e). Major promoter variations and haplotype *1 subtypes were subsequently analyzed for the association with ALL outcome. Lower EFS correlated with an A allele of G308A polymorphism (p=0.02) and with *1b haplotype (p=0.01). Higher mRNA levels were found in the carriers of *1b haplotype (p=0.005) and not for remaining haplotype *1 subtypes. So, the worse ALL outcome associated with haplotype *1 is actually determined by the subtype *1b. The study provides a new insight into DHFR regulatory polymorphisms defining more precisely event–predisposing DHFR variations.

Made available in DSpace on 2014-12-02T11:16:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-06-27Bitstream added on 2014-12-02T11:21:14Z : No. of bitstreams: 1 000693706.pdf: 2880957 bytes, checksum: 43d4b15dffaa346d602b5f57917996d4 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Amplamente utilizado no tratamento de vários tipos de câncer e na psoríase, o metotrexato (MTX) é um quimioterápico, estruturalmente análogo do ácido fólico, que apesar de sua eficácia apresenta uma série de efeitos adversos, sendo a hepatotoxicidade o mais grave. Atualmente os sistemas nanoestruturados líquido-cristalinos de fase lamelar estão sendo utilizados como dispositivos para liberação modificada de fármacos, sendo vantajosos na liberação tópica de várias substâncias, conforme suas características de interação com o estrato córneo e as outras camadas da pele, evitando assim efeitos adversos sistêmicos. Os objetivos deste trabalho foram desenvolver sistemas nanoestruturados líquido-cristalinos de fase lamelar, acrescidos de MTX, caracteriza-los do ponto de vista físico, realizar a análise estrutural das formulações, através de microscopia de luz polarizada (MLP), espalhamento de raios-X a baixo ângulo (SAXS) e suas propriedades reológicas, executar os testes de estabilidade preliminar (TEP) das formulações, validar o método analítico para a quantificação de MTX por CLAE e executar ensaios de liberação, permeação e retenção in vitro. As formulações preparadas a partir da mistura do poliéter funcional siloxano (Dow Corning® 5329) como tensoativo, com silicone fluido de co-polímero glicol (Dow Corning® 193C) como fase oleosa titulados em fase aquosa, composta por tampão fosfato de potássio monobásico 0,01M pH 7,4, apresentaram fases líquido-cristalinas do tipo lamelar, confirmados pelos ensaios de MLP e SAXS. Os TEPs evidenciaram que as formulações A, B e C mantiveram-se estáveis durante o período do estudo. Os estudos do comportamento reológico das formulações apresentaram-se como fluidos pseudoplásticos não–newtonianos tixotrópicos ... / Widely used in the treatment of some types of cancer, the methotrexate (MTX) is a chemotherapeutic, structurally analog of the folic acid, although its effectiveness, presents a series of adverse effect, being the most serious hepatotoxicity. Currently, the liquid crystal lamellar phase is being used of devices for modified release of drug demonstrated to be advantageous in the release topic of some substances, given to the characteristics of interaction with the stratum corneun and other layers of the skin, avoiding systemic adverse effects. The aims of this research had been to develop and to characterize liquid crystalline nanostructure systems of lamellar phase, increased of MTX of the physical point of view, also carry through the structural analysis of the formulations through by polarized light microscopy (PLM), small-angle X-ray scattering (SAXS) and rheological properties. Perform stability studies of the chosen formulations, validate the analytical method of quantification of MTX for High Performance Liquid Chromatography (HPLC) and carry through release assay, cutaneous permeation and skin retention in vitro for the chosen formulations. The formulations prepared by the mixture of polyether functional siloxane as surfactant, with silicone polyether copolymer as oily phase and phosphate buffer 0,01M pH 7,4 as aqueous phase demonstrating lamellar liquid-crystalline phases, confirmed by assays of PLM and SAXS. The stability studies showed that formulations A, B and C remained stable throughout the period of the study. The study of the rheological behavior of the formulations presented as not Newtonian pseudoplastic fluid and thixotropic ...