O adalimumabe (Humira®, Abbott) é um antagonista do Fator de Necrose Tumoral- alpha (TNF-alfa ). É aprovado para o tratamento de artrite reumatoide, espondilite anquilosante, doença de Crohn, psoríase crônica e artrite reumatoide juvenil. É um anticorpo monoclonal que contém apenas sequências humanas de peptídeos contra a molécula do Fator de Necrose Tumoral-alfa. Na literatura, relatos e série de casos sugerem que os antagonistas do Fator de Necrose Tumoral-alfa são úteis no tratamento da inflamação ocular, edema macular cistoide e secundário à uveíte e degeneração macular relacionada à idade. Entretanto, a administração sistêmica do adalimumabe pode gerar efeitos adversos graves. A fim de diminuir esses efeitos adversos e aumentar a concentração da medicação no segmento posterior do olho, uma possível opção é a injeção intravítrea. O objetivo do presente estudo foi avaliar a toxicidade do adalimumabe intravítreo nas diferentes doses para a retina de coelhos por meio de avaliação clínica (biomicroscopia e oftalmoscopia indireta), funcional (eletrorretinograma) e histopatológica (microscopia óptica e eletrônica). Foram utilizados 30 coelhos albinos da raça Nova Zelândia divididos em cinco grupos de seis coelhos. Injeções intravítreas foram realizadas nas seguintes concentrações de adalimumabe: 0,5mg/0,1ml, 1mg/0,1ml, 2,5mg/0,1ml, 5,0mg/0,1ml e 10mg/0,2ml e 0,1ml de solução salina balanceada (BSS) foi injetada nos olhos esquerdos dos grupos 1 e 2 para constituir o grupo controle. Foram realizadas biomicroscopia e fundoscopia e sinais de inflamação, infecção ou toxicidade foram observados durante duas semanas. O eletrorretinograma foi realizado antes do tratamento e após 14 dias da injeção intravítrea. Os animais foram sacrificados, foi feita a enucleação dos olhos, e o tecido para a avaliação histopatológica foi preparado. A injeção intravítrea de adalimumabe (Humira®) nas doses estudadas até 5mg (0,5mg, 1,0mg, 2,5mg, 5mg) não apresentou sinais clínicos, eletrorretinográficos e histopatológicos de toxicidade para a retina de coelhos a curto prazo. No grupo de 10mg, foram observados sinais inflamatórios leves em três dos seis olhos e houve diminuição da amplitude da onda a na resposta fotópica do ERG, não foram observadas alterações na microscopia óptica / Adalimumab is a fully human anti-TNF alpha monoclonal antibody consisting of 100% human sequences developed using phage display technology. It is currently FDA approved for the treatment of rheumatoid arthritis, ankylosing spondylitis, Crohns disease, moderate to severe chronic psoriasis, and juvenile idiopathic arthritis. Anti-TNF alpha drugs may be an effective therapy for cystoid macular edema associated with uveitis. Significant improvements in chronic diabetic macular edema and regression of CNV from AMD have also been documented in small published series after systemic treatment with TNF-alpha antagonists. However the systemic administration of these drugs can have serious side effects. Intravitreous injection would assure delivery of high concentrations of medication at the posterior segment with minimum side effects.The aim of this study was to evaluate the ocular toxicity of escalating doses of intravitreous adalimumab (Humira®) in the rabbit eye. Thirty New Zealand albino rabbits received intravitreous injections of 0.1ml of adalimumab 0.5 mg (6 eyes), 1mg (6 eyes), 2.5mg (6 eyes), 5mg (6 eyes) and 0.2ml was injected in the10mg (6 eyes) group. BSS (0,1ml) was injected in the left eye of the rabbits from the groups 1 and 2 to serve as control group. Slit lamp biomicroscopy, fundoscopy were carried out at baseline, day 7 and 14 following intravitreous injection while electroretinography (ERG) was carried out at baseline and day 14. Animals were euthanized on day 14 and histopathological examination of the eyes was performed. The tested doses of intravitreous adalimumab up to 5mg (0.5mg, 1.0mg, 2.5mg, 5mg) had no associated ocular short-term toxicity in rabbit eyes. The 10mg group showed mild inflammatory reaction in 3 out of 6 eyes and showed decrease in the a wave amplitude in the photopic response, light microscopy was normal

Wong Chun Lung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 202-225). / Abstracts in English and Chinese. / Thesis Committee --- p.ii / Abstract --- p.iii / 摘要 --- p.vi / Acknowledgements --- p.ix / Table of Contents --- p.x / List of Abbreviations --- p.xv / List of Figures --- p.xix / List of Tables --- p.xx iii / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- "Neurodegeneration, Inflammation and Gliosis" --- p.1 / Chapter 1.2 --- Anatomy of the CNS --- p.5 / Chapter 1.3 --- Astrocytes --- p.6 / Chapter 1.3.1 --- Morphology and Identification of Astrocytes --- p.6 / Chapter 1.3.2 --- Physiological Functions of Astrocytes in the CNS --- p.7 / Chapter 1.3.2.1 --- Induction of Blood-brain Barrier (BBB) --- p.7 / Chapter 1.3.2.2 --- Metabolism of Neurotransmitters --- p.9 / Chapter 1.3.2.3 --- Nursing Role of Astrocytes --- p.9 / Chapter 1.3.2.4 --- Immunological Functions of Astrocytes --- p.10 / Chapter 1.3.3 --- Neonatal Rat Cortical Astrocytes as In Vitro Model --- p.12 / Chapter 1.4 --- Cytokines in Brain Damage --- p.14 / Chapter 1.4.1 --- Lipopolysaccharides (LPS) --- p.16 / Chapter 1.4.2 --- Tumor Necrosis Factor-α (TNF-α) --- p.17 / Chapter 1.4.3 --- Interleukin-1 (IL-1) --- p.19 / Chapter 1.4.4 --- Interleukin-6 (IL-6) --- p.20 / Chapter 1.4.5 --- Interferon-γ (IFN-γ) --- p.21 / Chapter 1.5 --- Cytokines-induced Signaling Cascade --- p.22 / Chapter 1.5.1 --- TNF Receptors --- p.23 / Chapter 1.5.2 --- Ca2+ --- p.25 / Chapter 1.5.3 --- MAPK --- p.26 / Chapter 1.5.4 --- PICA --- p.27 / Chapter 1.5.5 --- NFkB --- p.29 / Chapter 1.6 --- Glucose Metabolism in the Brain and Glucose Transporters --- p.31 / Chapter 1.6.1 --- Glucose Transporters in the Brain --- p.32 / Chapter 1.6.2 --- Glucose Transporters in Brain Damage --- p.34 / Chapter 1.7 --- Ascorbic Acid Metabolism in the Brain --- p.36 / Chapter 1.8 --- Aim and Scope of this Project --- p.39 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials / Chapter 2.1.1 --- Neonatal Sprawley 一Dawley Rats --- p.43 / Chapter 2.1.2 --- Plain Dulbecco Modified Eagle Medium ´ؤ Formula 12 (pDF12) --- p.43 / Chapter 2.1.3 --- Complete DF-12(cDF12) --- p.43 / Chapter 2.1.4 --- Phosphate Buffered Saline (PBS) --- p.44 / Chapter 2.1.5 --- Hank's Buffer (HSB) --- p.44 / Chapter 2.1.6 --- D/L-Homocysteine Buffer --- p.44 / Chapter 2.1.7 --- "LPS, Cytokines and Pentoxifylline" --- p.45 / Chapter 2.1.8 --- Specific TNF Receptor Agonist: TNF antibodies --- p.45 / Chapter 2.1.9 --- Calcium Modulators --- p.45 / Chapter 2.1.10 --- PKA Modulators --- p.46 / Chapter 2.1.11 --- NFkB Inhibitors --- p.47 / Chapter 2.1.12 --- MAPK Inhibitors --- p.47 / Chapter 2.1.13 --- β-Adrenergic Receptor Modulators --- p.47 / Chapter 2.1.14 --- Reagents for RNA and Protein Isolation --- p.48 / Chapter 2.1.15 --- Reagents for Reverse Transcription-Polymerase Chain Reaction (RT-PCR) --- p.48 / Chapter 2.1.16 --- Reagents for DNA Electrophoresis --- p.49 / Chapter 2.1.17 --- Reagents for Real-time PCR --- p.51 / Chapter 2.1.18 --- Reagents for Western Blotting --- p.51 / Chapter 2.1.19 --- Reagents for MTT Assay --- p.51 / Chapter 2.1.20 --- Reagents for 3H-Thymidine Incorporation Assay --- p.52 / Chapter 2.1.21 --- Reagents for Glucose Uptake Assay --- p.52 / Chapter 2.1.22 --- Reagents for Ascorbic Acid Accumulation Assay --- p.53 / Chapter 2.1.23 --- Reagents for Immunostammg --- p.53 / Chapter 2.1.24 --- Other Chemicals and Reagents --- p.53 / Chapter 2.2 --- Methods / Chapter 2.2.1 --- Preparation of Primary Cultured Rat Astrocytes --- p.55 / Chapter 2.2.2 --- Measuring Cell Viability: MTT Assay --- p.56 / Chapter 2.2.3 --- Measuring Cell Proliferation: 3H Thymidine Incorporation Assay --- p.57 / Chapter 2.2.4 --- Measuring Glucose Uptake: Zero-trans Glucose Uptake Assay --- p.58 / Chapter 2.2.5 --- Measuring Ascorbic Acid Accumulation --- p.60 / Chapter 2.2.6 --- Total Protein Extraction --- p.61 / Chapter 2.2.7 --- Western Blotting --- p.62 / Chapter 2.2.8 --- Immunostaining --- p.64 / Chapter 2.2.9 --- Isolation of RNA --- p.64 / Chapter 2.2.10 --- Measurement of RNA Yield --- p.65 / Chapter 2.2.11 --- RNA Gel Electrophoresis --- p.66 / Chapter 2.2.12 --- Reverse Transcription (RT) --- p.66 / Chapter 2.2.13 --- Polymerase Chain Reaction (PCR) --- p.67 / Chapter 2.2.14 --- Separation of PCR Products by Agarose Gel Electrophoresis --- p.67 / Chapter 2.2.15 --- Quantization of PCR Products and Western Blotting --- p.68 / Chapter 2.2.16 --- Real-time PCR --- p.68 / Chapter Chapter 3 --- Results / Chapter 3.1 --- Role of Calcium Ions (Ca2+) in TNF-α-induced Astrocyte Proliferation --- p.70 / Chapter 3.1.1 --- Effects of Changes of Extracellular Ca2+ on Astrocyte Viability --- p.72 / Chapter 3.1.2 --- Effects of Other Divalent Ions on Astrocyte Viability --- p.74 / Chapter 3.1.3 --- Effects of Changes of Intracellular Ca2+ on Astrocyte Viability --- p.78 / Chapter 3.1.4 --- Role of Ca2+ on TNF-α-mduced Proliferation in Astrocytes --- p.85 / Chapter 3.1.5 --- Role of Other Divalent Ions on tnf-α-mduced Proliferation in Astrocytes --- p.90 / Chapter 3.2 --- Effect of Cytokines on Glucose Uptake in Rat Astrocytes --- p.95 / Chapter 3.2.1 --- Basal level of Glucose Uptake in Astrocytes and Effects of Cytokines on Glucose Uptake in Astrocytes --- p.95 / Chapter 3.2.2 --- Signaling Cascade of LPS- and TNF-α-induced Glucose Uptake in Astrocytes --- p.120 / Chapter (A) --- TNFR Subtypes Mediating TNF-a-induced Glucose Uptake --- p.121 / Chapter (B) --- MAPK --- p.125 / Chapter (C) --- PKA --- p.133 / Chapter (D) --- NFkB --- p.139 / Chapter (E) --- Other Mechanisms / Signalling molecules --- p.150 / Chapter (1) --- Interaction with β-Adrenegic Mechanism / Chapter (2) --- Role of cGMP --- p.154 / Chapter (3) --- Effect of Mg2+ on LPS- / TNF-α- induced Glucose Uptake in Astrocytes --- p.156 / Chapter (4) --- Possible Involvement of IGF-1 System --- p.160 / Chapter 3.2.3 --- Summary --- p.163 / Chapter 3.3 --- Effects of LPS and Cytokines on AA Accumulation in Astrocytes --- p.164 / Chapter Chapter 4 --- Discussion / Chapter 4.1 --- Role of Calcium ions (Ca2+) in TNF-α-induced Astrocyte Proliferation --- p.177 / Chapter 4.1.1 --- Drastic Changes in Extracellular Ca2+ Caused Astrocyte Death --- p.178 / Chapter 4.1.2 --- Extraordinary Role of Ca2+ in Astrocytes Survival --- p.178 / Chapter 4.1.3 --- Elevation of [Ca2+]i Reduced Astrocyte Viability --- p.180 / Chapter 4.1.4 --- Failure of Verapamil to Block TNF-α-induced Astrocyte Proliferation --- p.182 / Chapter 4.2 --- Hypothesis for the Relationship between Cytokines and Energy Metabolism --- p.185 / Chapter 4.2.1 --- Mechanism and Signaling Cascade of the Elevated Glucose Uptake --- p.186 / Chapter 4.2.2 --- Increased Glucose Uptake by Cytokines: Friend or Foe? --- p.191 / Chapter 4.2.3 --- Depletion of AA Pool by LPS --- p.194 / Chapter 4.2.4 --- Possible Bedside Application of the Findings --- p.195 / Chapter 4.3 --- Prospects of This Study and Concluding Remarks --- p.197 / Appendix --- p.201 / References --- p.202

Made available in DSpace on 2014-06-11T19:22:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-02-21Bitstream added on 2014-06-13T18:08:04Z : No. of bitstreams: 1 cardoso_crp_me_arafcf.pdf: 859101 bytes, checksum: dc8d0d66975b1493a8b7a5a2830d3074 (MD5) / Universidade Estadual Paulista (UNESP) / Considerando a pesquisa de novas moléculas farmacologicamente ativas, orientada pelo uso tradicional das plantas com finalidade terapêutica, o presente trabalho avaliou os efeitos genotóxicos e de ativação do sistema imunológico em duas espécies de plantas do gênero Byrsonima: Byrsonima crassa e Byrsonima intermedia. Essas espécies são plantas de uso popular, pertencentes à flora do Cerrado Brasileiro, utilizadas pela população para disfunções gástricas e diarréias. Foram realizados ensaios para a caracterização da mutagenicidade dos extratos, através do Teste de Ames (TA), utilizando-se linhagens geneticamente modificadas de Salmonella typhimurium e de ativação da resposta imune celular, utilizando-se cultura de macrófagos de exsudato peritoneal (PEC) de camundongos Swiss, avaliando a liberação celular de Fator de Necrose Tumoral-alfa (TNF-α) e Óxido nítrico (NO), importantes moduladores da resposta inflamatória. Foi verificado que somente o extrato metanólico de B.crassa apresentou atividade mutagênica positiva na linhagem TA98, com e sem ativação metabólica. Analisando-se as frações aquosa e acetato de etila, verificou-se que o composto provavelmente responsável pela atividade mutagênica do extrato metanólico está presente na fração acetato. Testes realizados com os compostos isolados dessa fração (catequina, galato de metila, querecetina -3-O-β-Dgalactopiranosídeo, quercetina-3-O-α-arabinopiranosídeo e amentoflavona) revelaram ação mutagênica da amentoflavona e indícios de mutagenicidade da quercetina arabinopiranosídeo. Nos ensaios imunológicos, verificou-se que os extratos de B.crassa e B.intermedia promoveram estímulo da liberação de NO por macrófagos de camundongos em níveis reduzidos, assim como a fração aquosa do extrato metanólico de B.crassa. A fração acetato de etila do extrato... / Considering the research of active new molecules, leaded by the traditional use of plants with therapeutic proposal, this paper tested the genotoxic effects and the activation of the immune system in two species of Byrsonima: Byrsonima crassa and Byrsonima intermedia. These are popular use species that belong to the Brazilian Cerrado, utilized by population to gastric dysfunctions and diarrheas. Many assays were done to give the characterization of mutagenicity of the extracts, through the Ames test (TA) using a line of genetic changed Salmonella typhimurium and the activation of the cellular response, using a culture of peritoneal macrophages from Swiss mice, to test the cell liberation of the Tumor Necrosis Factor-apha (TNF-a) and Nitric Oxide (NO), important modulators on the inflammation response. It was verified that only the methanolic extract of B.crassa presented positive mutagenic activity in the line TA98, with and without metabolic activation. Analyzing the watering and ethyl acetate portions, it was checked that the substance that probably is responsible by the mutagenic activity of the methanolic extract is present on the ethyl acetate portion. Assays achieved with isolated compounds from this portion - (+)-catechin, methyl galate, quercetin-3-O-ß-D-galactopyranoside, quercetin-3-O-a-Larabinopyranoside and amentoflavone - discovered mutagenic activity of amentoflavone and mutagenicity signals of quercetin-3-O-a-L-arabinopyranoside. On this immunologic assays, it was observed that the B.crassa and B.intermedia extracts did not promote the stimulation of leaving NO and TNF-a by macrophages of mouse, just live the watering portion from B.crassa methanolic extract. The ethyl acetate B.crassa portion presented stimulated and relevant activity to the liberation of NO and TNF-a. (Complete abstract, click electronic address below).

Ngan, Kit Shan. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 176-187). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese.

Thesis (Ph. D.)--West Virginia University, 2003. / Title from document title page. Document formatted into pages; contains xv, 190 p. : ill. (some col.). Includes abstract. Includes bibliographical references.

Oxidative stress and inflammation are important factors involved in the progression of heart failure. An important cytokine produced during myocardial infarction (MI) is tumor necrosis factor alpha (TNF-α). TNF-α may induce oxidative stress, cell damage, apoptosis and cardiac dysfunction. Considering the anti-inflammatory and anti-oxidant properties of extra-virgin olive oil and its major component (80%) oleic acid (OA), and their benefits to the cardiovascular system, we hypothesized that the negative effects of TNF-α in the pathogenesis of heart failure will be mitigated by olive oil consumption. This hypothesis was tested by examining the effects of a special diet supplemented with 10% olive oil, in coronary artery ligated animal model of MI. Corn oil (10%) supplementation was used as a control for matching caloric intake. Animals in the sham and ligated groups fed regular chow, olive oil, and corn oil were studied at 4 and 16 weeks post myocardial infarction (PMI). Mortality, diet consumption, weight gain and conduction system abnormalities were comparable among all ligated groups. Echocardiography showed that MI deteriorated cardiac function, and olive oil restored the function. At 16 weeks PMI, only corn oil fed groups showed significant increase in both total cholesterol and HDL. Corn oil was not able to offer protection to the heart, suggesting that the beneficial effects of olive oil are not due to increased caloric intake or increased HDL. MI increased myocardial TNF-α, oxidative stress, lipid peroxidation, pro-apoptotic protein expression (Bax, cleaved Caspase 3, cleaved PARP, TGFβ, Bnip3), cytochrome C release, MAP kinase activation (p38, JNK) and decreased anti-apoptotic protein Bcl-xL expression at both 4 and 16 weeks PMI, and these changes were modulated by olive oil. In order to further test the central role of TNF-α PMI, we examined the possible miti-gation of TNF-α induced changes by OA in isolated adult rat cardiomyocytes. TNF-α in-creased oxidative stress, cell damage, cell death, and apoptosis, while OA treatment miti-gated these TNF-α induced effects. We concluded that TNF-α is implicated in the progression of heart failure subsequent to MI and that OA in olive oil may prevent this progression, through its anti-oxidant, anti-inflammatory, anti-hypertensive, and inotropic effects.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 5 uppsatser.

Dissertation (Ph.D.)--University of South Florida, 2005. / Includes vita. Includes bibliographical references (leaves 179-187). Also available online.

Thesis (M.Sc.)--Memorial University of Newfoundland, Faculty of Medicine, / Typescript. Bibliography: leaves 44-68.

Thesis (M.S.)--University of Florida, 2004. / Typescript. Title from title page of source document. Document formatted into pages; contains 49 pages. Includes Vita. Includes bibliographical references.