Made available in DSpace on 2016-01-26T12:51:29Z (GMT). No. of bitstreams: 1 aneliserusso_dissert.pdf: 4806118 bytes, checksum: 0ae19d04c260950faf6788fde7ad65a0 (MD5) Previous issue date: 2011-05-24 / Some individuals may be present increased risk of developing cancer due to differences in biometabolism. Polymorphisms in xenobiotic metabolizing genes, such as family members of Cytochrome P450 (CYP), of Glutathione-S-Transferases (GSTs) and Microsomal Epoxide Hydrolase (mEH) show association with the carcinogenesis of head and neck. Objective: Identify the frequency of genes polymorphisms CYP1A1 (CYP1A1*2A and CYP1A1*2C), CYP2E1 (CYP2E1*5B and CYP2E*6), GSTT1 (null genotype), GSTM1 (null genotype), GSTP1 (A313G and C341T) and mEH (Tyr113His and His139Arg) in patients with head and neck cancer and in individuals with no cancer history (controls), to identify susceptibility biomarkers of this type of cancer. Methods: included 1,100 individuals, 375 patients with pathological diagnosis of squamous cell carcinoma of head and neck cancer and 725 controls. The variables analyzed were: age, sex, alcohol and tobacco, the occurrence primary site and progression of tumor. Genotyping of polymorphisms was performed by Polymerase Chain Reaction - Length Polymorphism, Restriction Fragment (PCR-RFLP), Real-Time PCR (RT-PCR) and multiplex PCR. Datas were evaluated by chi-square, univariate and multiple logistic regression. Results: Advanced age, smoking habits and alcohol consumption were predictors for the development of the head and neck tumors. The polymorphisms CYP2E1*5B (CYP2E1-PstI) and GSTP1 A313G were associated with this disease in univariable analysis. In multirariable analysis the interaction among CYP1A1*2C polymorphism with the female gender (OR= 0.10; 95% CI=0.01-0.72; p< 0.05) and alcohol no-habit (OR= 0.21; 95% CI=0.07-0.67; p< 0.05), mEH His139Arg and alcohol habit (OR= 0.49; 95% CI=0.27-0.90; p< 0.05), CYP2E1* 5B and habits tobacco and alcohol and &#8805;49 age (OR = 4.10; CI 95% 2.44-6.89; p < 0.001; OR = 1.93; CI 95% 1.18-3.16; p=0.0084; OR=9,10; CI 95% 5,86-14,14; p< 0,001, respectively) and GSTP1 A313G and tobacco and alcohol habits, &#8805;48 agea and male gender (OR=4.21; IC 95% 2.71-6.55; p<0.0001; OR=1.65; IC 95% 1.07-2.55; p=0.023; OR=12.37; IC 95% 7.89-19.38; p<0.0001, respectively); decrease the head and neck cancer risk; while CYP1A1*2A and tobacco and alcohol no-habits (OR= 2.84; 95% CI=1.01-7.97; p< 0.05; OR= 2.43; 95% CI=1.00-5.87; p< 0.05, respectively) increase this risk. The CYP2E1*5B polymorphism and tobacco no-habit (OR= 3.75; 95% CI=1.25-11.23; p< 0.05) also increases the risk for this disease. Showed high frequency of the polymorphisms CYP1A1*2C (OR= 2.48; 95% CI=1.00-6.20; p < 0.05) GSTT1 (OR= 3.35; 95% CI=1.67-6.72; p < 0.05) in patients group with primary tumors of larynx; while those who have the pharynx as the primary site GSTT1 (OR=0.29; 95% CI=0.12-0.71; p < 0.05) was less frequent. CYP1A1*2A (CYP1A1-MspI) polymorphism was associated with lymph node involvement and increased risk for cancer (OR =2.45; 95% CI= 1.07-5.64). Conclusion: Our data demonstrate that the interaction between polymorphisms in genes that encode enzymes involved in xenobiotic metabolism and the demographic and risk factors may modulate the development of head and neck cancer. / Alguns indivíduos podem apresentar risco aumentado de desenvolver o câncer devido às diferenças no biometabolismo. Polimorfismos em genes metabolizadores de xenobióticos, tais como os membros da família do Cytocromo P450 (CYP), das Glutatião-S-Transferases (GSTs) e Epóxido Hidrolase Microssomal (mEH) mostram associação com a carcinogênese de cabeça e pescoço. Objetivo: Identificar a frequência de polimorfismos dos genes CYP1A1 (CYP1A1*2A e CYP1A1*2C), CYP2E1 (CYP2E1*5B e CYP2E*6), GSTT1 (genótipo nulo), GSTM1 (genótipo nulo), GSTP1 (A313G e C341T) e mEH (Tyr113His e His139Arg) em pacientes com câncer de cabeça e pescoço e em indivíduos sem história de neoplasia (controles), visando identificar biomarcadores de suscetibilidade para este tipo de câncer. Casuística e Métodos: Foram incluídos no estudo 1.100 indivíduos, 375 pacientes com diagnóstico patológico de carcinoma de células escamosas de cabeça e pescoço e 725 controles. As variáveis analisadas foram: idade, sexo, consumo de álcool e tabaco, sítio primário de ocorrência de tumor e evolução da doença. A genotipagem dos polimorfismos foi realizada por Reação em Cadeia da Polimerase Polimorfismos de Comprimentos de Fragmento de Restrição (PCR-RFLP), PCR em Tempo Real (PCR-RT) e PCR multiplex. Os dados foram avaliados por Qui-quadrado e Regressão logística univariada e múltipla. Resultados: Idade acima de 49 anos, hábitos tabagista e etilista foram preditores para o desenvolvimento de neoplasias de cabeça e pescoço. Os polimorfismos CYP2E1*6 (CYP2E1-DraI) e GSTP1 A313G foram associados com esta doença na análise univariada. Na análise múltipla, a interação entre polimorfismo CYP1A1*2C com o gênero feminino (OR= 0,10; 95% CI=0,01-0,72; p< 0,05) e hábito não etilista (OR= 0,21; 95% CI=0,07-0,67; p< 0,05), mEH His139Arg e hábito etitlista (OR= 0,49; 95% CI=0,27-0,90; p< 0,05), CYP2E1* 5B e hábitos tabagista e etilista e idade &#8805;49 (OR = 4,10; CI 95% 2,44-6,89; p < 0,001; OR = 1,93; CI 95% 1,18-3,16; p = 0,0084; OR = 9,10; CI 95% 5,86-14,14; p < 0,001, respectivamente) e GSTP1 A313G e hábitos tabagista e etilista, idade &#8805;48 anos e gênero masculino (OR=4,21; IC 95% 2,71-6,55; p<0,0001; OR=1,65; IC 95% 1,07-2,55; p=0,023; OR=12,37; IC 95% 7,89-19,38; p<0,0001, respectivamente); diminui o risco de câncer de cabeça e pescoço; enquanto CYP1A1*2A e hábitos não tabagista e não etilista (OR= 2,84; 95% CI=1,01-7,97; p< 0,05; OR= 2,43; 95% CI=1,00-5,87; p< 0,05, respectivamente) aumentam este risco. O polimorfismo CYP2E1*5B e hábito não tabagista (OR= 3,75; 95% CI=1,25-11,23; p< 0,05) também aumenta o risco para esta doença. Foi observada uma alta frequência dos polimorfismos CYP1A1*2C (OR= 2,48; 95% CI=1,00-6,20; p < 0,05) GSTT1 (OR= 3,35; 95% CI=1,67-6,72; p < 0,05) no grupo de pacientes com tumores primários de laringe; enquanto naqueles que tem a faringe como sítio primário o polimorfismo GSTT1 (OR=0,29; 95% CI=0,12-0,71; p < 0,05) foi menos freqüente. O polimorfismo CYP1A1*2A foi associado com o envolvimento de linfonodo e risco aumentado para o câncer (OR=2,45; 95% CI=1,07-5,64). Conclusão: Nossos dados demonstram que a interação entre os polimorfismos em genes que codificam enzimas envolvidas no metabolismo de xenobióticos e os fatores demográficos e de risco podem modular o desenvolvimento do câncer de cabeça e pescoço.

Made available in DSpace on 2015-07-22T20:49:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-09-24. Added 1 bitstream(s) on 2015-08-11T03:26:00Z : No. of bitstreams: 1 Publico-120.pdf: 952558 bytes, checksum: 1bf84fc7ff3623ea692faec0f6256685 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Objetivo: investigar a associação do polimorfismo do CYP 17 com leiomioma uterino, uma vez que os hormônios esteróides têm papel na patogênese dessa doença e a enzima codificada pelo CYP 17 está envolvida na biossíntese desses hormônios. Casuística e métodos: foram comparadas, em estudo caso-controle, 121 mulheres com leiomioma uterino sintomático que se submeteram a tratamento cirúrgico (Caso) e 120 mulheres na peri ou na pós-menopausa, sem diagnóstico prévio ou atual de leiomioma (Controle). Os grupos foram analisados quanto à presença do polimorfismo do gene CYP 17. A genotipagem do CYP 17 foi realizada por meio da reação em cadeia da polimerase com polimorfismo de comprimento de fragmento de restrição (PCR-RFLP) com a enzima de restrição Mspa1. Nas mulheres com a doença, o DNA foi proveniente de material de histerectomia e, nos controles, por meio de sangue coletado em punção venosa periférica. Para a análise estatística, foi realizado o teste do qui-quadrado seguido de regressão logística. O nível de significância adotado foi de 5% (p<0,05) e o intervalo de confiança foi de 95% (95% IC). Finalmente, confirmou-se o equilíbrio de Hardy-Weinberg para as amostras. Resultados: comparando-se mulheres com leiomiomas uterino e controles, notamos ausência de diferença significativa na distribuição do genótipo do CYP 17 (p 0,165) entre os dois grupos. Conclusões: não houve associação do polimorfismo do CYP 17 com a ocorrência de leiomioma uterino na população estudada. / Objective: Uterine leiomyoma is the most common pelvic tumor in women of reproductive age. It has been well established that endogenous sex hormones are involved in the pathogenesis of the disease, and polymorphisms in genes encoding for enzymes that act in the steroid hormones metabolism, as the CYP17, may therefore play a role in the genesis of fibroids. Variations in this gene have been thought to be candidates influencing the susceptibility to hormone-related diseases. A single nucleotide polymorphism (T→C) [rs1042386] in the promoter region of CYP17 is speculated to alter its transcription. The present study was conducted to investigate the association between this polymorphism and the presence of uterine leiomyoma in Brazilian women. Methods: Genotyping of the CYP17 was performed in 121 uterine fibroid patients and 120 unaffected women using polymerase chain reaction and restriction fragment length polymorphism analysis. Results: No significant difference in the CYP17 genotype distribution was noted between cases and controls (p=0.165) Conclusion: These findings suggest that the CYP17 gene polymorphism studied is unlikely to be associated with risk for uterine leiomyoma in Brazilian women. / FAPESP: 03/04533-1 / TEDE / BV UNIFESP: Teses e dissertações

La bio-transformation naturelle des médicaments peut produire des métabolites toxiques; l’identification de ces métabolites est essentielle dans la stratégie de choix de molécules thérapeutiques. En appliquant les technologies de fermentation en bioréacteur des cellules hétérologues (souches d’E. coli recombinantes exprimant une iso-enzyme de cytochrome P450 humain avec la réductase humain), la bioconversion du substrat (principe actif) en ses métabolites de dégradation, a été réalisée à grande échelle (& / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

Substanzen, die durch ihr hormonell wirksames Potenzial mit dem Hormonsystem interagieren und adverse Effekte auf die Reproduktion von Invertebraten und Vertebraten ausueben koennen, erlangten in den letzten Jahrzehnten große Aufmerksamkeit. Viele dieser Substanzen reduzieren die Fertilitaet oder die Fekunditaet, fuehren zu Abnormalitaeten in der Ontogenese oder im Verhalten der Tiere und haben Einfluss auf die Geschlechterverhaeltnisse. In der vorliegenden Arbeit wurden verschiedene Aspekte dieses Themengebietes bearbeitet. Das in Europa endemisch vorkommende Rotauge (Rutilus rutilus), ein Sueßwasserfisch, wurde als Modelltier fuer den Nachweis von (anti)androgenen Effekten auf aquatisch lebende Organismen etabliert. Zum Nachweis der (anti)androgenen Wirkmechanismen wurden die Tiere mit Modellsubstanzen aus drei verschiedenen Gruppen exponiert. Aus der Gruppe der Substanzen mit potenziell androgener Wirkung wurden Triphenylzinn (TPT) und Methyltestosteron (MT) verwendet, aus der Gruppe der Antiandrogene Vinclozolin (VIN) und Cyproteronazetat (CYP) und aus der Gruppe der Aromatasehemmer, und somit potenziell androgener Wirkung, Letrozol (LET) und Fenarimol (FEN). Feedbackmechanismen auf die Hypothalamus-Hypophysen-Gonaden-Achse (mRNA-Expression des Luteinisierenden Hormons, des Follikel stimulierenden Hormons und der Aromatase), mRNA-Expression potentieller Biomarker in der Leber (Androgen-Rezeptor-mRNA, Oestrogen-Rezeptor-mRNA), Sexsteroidspiegel im Blutplasma (17beta-Oestradiol und 11-keto-Testosteron), Enzymaktivitaeten im Gehirn (Aromatase), Histologie der Gonaden, Totallaenge, Gewicht und Geschlechterverteilung wurden als Endpunkte analysiert, um adverse Effekte auf die Reproduktionsbiologie von R. rutilus zu zeigen. Die untersuchten Endpunkte eigneten sich sehr gut zum Nachweis verschiedener Wirkmechanismen. / Substances that are able to interact with the endocrine system and cause adverse effects on the reproduction of invertebrates and vertebrates have gained much attention over the last few decades. Many of these substances reduce fertility or fecundity, lead to developmental abnormalities or abnormalities in the behaviour of animals and have an impact on sex ratios. The present study examines various aspects of these topics. The roach (Rutilus rutilus), a freshwater fish endemic in Europe, was established as a model animal for the detection of (anti)androgenic effects on aquatic organisms. For examination of the (anti)androgenic action, the animals were exposed to model compounds from three different groups: triphenyltin (TPT) and methyltestosterone (MT) from the group of substances with potentially androgenic effect, vinclozolin (VIN) and cyproteronacetate (CYP) from the group of antiandrogens, and letrozol (LET) and fenarimol (FEN) from the group of aromatase inhibitors, which thus have a potentially androgenic effect. Feedback mechanisms on the hypothalamus-pituitary-gonad-axis (mRNA expression of luteinising hormone, follicle stimulating hormone and aromatase), mRNA expression of potential biomarkers in the liver (androgen receptor mRNA, oestrogen receptor mRNA), steroid levels in the blood plasma (17beta-oestradiol and 11-ketotestosterone), enzyme activity in the brain (aromatase), histology of the gonads, total length, weight and sex ratios were analysed as endpoints to show adverse effects on the reproductive biology of R. rutilus. The studied endpoints are suitable for the detection of different modes of action. The histological examination of the gonads proved to be especially sensitive with the exposure to AACs to resulting in fundamental adverse damages to the gonads. It was ascertained that - in the early stages of ontogeny - androgens play as crucial of a role in the development of the gonads as previously attributed primarily to oestrogens.

Disease- and drug-related bone disorders are rapidly increasing in the population. The drugs which are used for the treatment of neurodegenerative diseases and metabolic derangements, may have negative or positive effects on bone tissues. In the first study, the possible side-effects of Carbamazepine and epileptic seizures on bone structure and composition were investigated by FTIR and synchrotron-FTIR microspectroscopy, AFM and micro- and nano-hardness analysis. The effects on the blood parameters, bone turnover and vitamin D metabolism were also investigated by ELISA and western blot analysis. The current study provides the first report on differentiation of the effects of both epileptic seizures and AED therapy on bones. Besides Carbamazepine treatment, seizures also caused a decrease in the strength of bone. The biochemical data showed that both the epileptic and drug-treated groups decreased vitamin D levels by increasing the vitamin D catabolism enzyme / 25-hydroxyvitamin D-24-hydroxylase. In the second study, the possible pleiotropic (positive) effects of cholesterol lowering drug / Simvastatin on bones were investigated by ATR-FTIR spectroscopy. The current study provides the first report on dose-dependent effects of simvastatin on protein structure and lipid conformation of bones. ATR-FTIR studies showed that although both high and low dose simvastatin strengthen bones, low dose simvastatin treatment is much more effective in increasing bone strength. Neural network analysis revealed an increased antiparallel and aggregated beta sheet and random coil in the protein secondary structure of high dose group implying a protein denaturation. Moreover, high dose may induce lipid peroxidation which limit the pleiotropic effects of high dose treatment on bones. This study clearly demonstrated that using low dose simvastatin is safer and more effective for bone health than high dose simvastatin treatment.

Bakgrund: Det finns läkemedel som ger ökad risk för vitamin D-brist. En kartläggningsstudie av hur patientgrupper med olika riskläkemedel hanteras i Landstinget i Kalmar län har inte gjorts tidigare. En sådan studie är önskvärd, för att få en nulägesanalys och öka medvetenheten bland hälso-och sjukvårdspersonal om hur läkemedel påverkar D-vitaminstatus. Syfte: Syftet var att undersöka omfattningen av ordinerade riskläkemedel hos samtliga patienter, samt hos patientgruppen ≥75år och hur dessa handläggs avseende ordination av supplementering (samtidig behandling med läkemedel innehållande vitamin D3 / kalcium och vitamin D3,), provtagning och analysresultat avseende kalcidiol i serum. Metod: Riskläkemedel för vitamin D-brist identifierades. Inklusionskriterier för studien var patienter med de fördefinierade riskläkemedlen med ordinationer under 2012 och/eller 2013. Populationen som ingick i studien var patienter i Landstinget i Kalmar län. Data hämtades från patientdatasystemet Cambio Cosmic. Utifrån dessa data analyserades förekomst av supplementering, provtagning för S-25(OH)D kalcidiol och analyssvar. Materialet strukturerades efter respektive läkemedel avseende kön och ålder (≥75 år.) Resultat: 9118 individer ordinerades något av riskläkemedelen orlistat, sevelamer, kolestyramin, kolestipol, efavirenz, prednison, prednisolon, fenytoin, fenobarbital eller karbamazepin. 31% av patienterna som ordinerats riskläkemedel var ≥75 år. Totalt behandlades 22% med supplementationspreparat av de som ordinerats riskläkemedel. I åldersgruppen ≥75 år behandlades 43% med supplement. 61% av de supplementerade patienterna var ≥75 år. Totalt provtogs enbart 4,1% avseende kalcidiol av de som ordinerats riskläkemedel. Av dessa hade 37% av de provtagna en bristnivå av S-25(OH)D (≤50 nmol/L) vid första provtillfället. 57% av de som visade brist supplementerades. Endast 8,3% av de som hade en brist följdes upp med ytterligare en eller flera provtagningar. Av patienterna ≥75 år provtogs 4,1% av patienterna i åldersgruppen. Detta innebär att 31% av de som provtogs var ≥75år. 39% led brist. Av de patienter ≥75 år som hade led brist supplementerades 65% av individerna. Konklusion: Det finns förbättringsutrymme för implementering av kunskap kring riskläkemedel i Landstinget i Kalmar län. / Introduction: Some drugs increase the risk for vitamin D deficiency. To date, no survey has been performed in Kalmar County about how patients medicated with risk pharmaceuticals are handled. Such a survey would be desirable in order to increase knowledge among healthcare workers about how substances interfere with vitamin D status. Aim: The aim was to examine the extent to which risk pharmaceuticals are prescribed among all patients and among patients aged ≥75, and to investigate how these patients are treated with respect to supplementation (drugs containing vitamin D3 / Calcuim and vitamin D3), blood sampling and analysis of serum calcidiol levels. Method: Drugs interfering with vitamin D were identified. Inclusion criteria for the survey were patients that had been prescribed the pre-defined risk pharmaceuticals during 2012 and/ or 2013. The surveyed population was residents in Kalmar County. Data were collected from the patient database Cambio Cosmic. From these data analyzes were made about the occurrence of: supplementation, sampling of S-25(OH)D calcidiol and test results. The collected data were arranged by substance, gender and age (elderly aged ≥75). Results: 9118 patients were prescribed at least one of the risk pharmaceuticals; orlistat, sevelamer, cholestyramine, colestipol, efavirenz, prednisol, prednison, phenytoin, phenobarbital or carbamazepine. 31% of these were aged ≥75. Overall, 22% of the patients were prescribed supplements. Out of these, 61% were elderly. Among the patients that had been prescribed risk pharmaceuticals a minority, 4,1% of the patients were sampled for calcidiol. 37% of these had deficiency in S-25(OH)D (≤50 nmol/L). 8.3% of patients with a vitamin D deficiency were sampled more than once. Out of the patients with deficiency 57% were treated with supplements. Out of the elderly patients prescribed risk pharmaceuticals, 4.1% of the patients were tested for calcidiol. This means that 31% of the tested patients were aged ≥75 and 39% of these had a deficiency. Out of the elderly patients with deficiency, 65% were treated with supplements. Conclusion: In Kalmar County, much can be done in order to better implement the existing knowledge about drugs that increase the risk for vitamin D deficiency. / Betydelsen av bra D-vitaminstatus för äldres hälsa – Hur stor är förekomsten av D-vitaminbrist hos äldre i Kalmar län och hur påverkar läkemedel D-vitaminstatus