Submitted by Simone Fortunato (simone.fortunato@conveniado.bahia.fiocruz.br) on 2017-10-04T13:49:03Z No. of bitstreams: 1 2017 Rifkath_19 de setembro colorido.pdf: 1577388 bytes, checksum: db9deae3ad793c3fb4c568df25016401 (MD5) / Approved for entry into archive by Uillis de Assis Santos (uillis.assis@ufba.br) on 2017-10-04T14:02:02Z (GMT) No. of bitstreams: 1 2017 Rifkath_19 de setembro colorido.pdf: 1577388 bytes, checksum: db9deae3ad793c3fb4c568df25016401 (MD5) / Made available in DSpace on 2017-10-04T14:02:02Z (GMT). No. of bitstreams: 1 2017 Rifkath_19 de setembro colorido.pdf: 1577388 bytes, checksum: db9deae3ad793c3fb4c568df25016401 (MD5) / RESUMO INTRODUÇÃO: Os níveis séricos de bilirrubina encontram-se frequentemente aumentados em pacientes com anemia falciforme, já acometidos por hemólise exacerbada, favorecendo a agravação do quadro clinico e a ocorrência de litíases biliares. OBJETIVO: O objetivo foi analisar a influência do polimorfismo A(TA)nTAA no promotor do gene UGT1A1 (rs8175347) e de dois polimorfismos no gene BCL11A (rs1427407 e rs7606173), sobre alguns parâmetros laboratoriais e sobre a resposta à hidroxiureia em pacientes pediátricos com anemia falciforme. MATERIAL E MÉTODOS: Amostras de sangue total e soro foram utilizadas para a realização das análises hematológicas e bioquímicas. O DNA foi extraído a partir de amostras de sangue periférico, e os fragmentos contendo os polimorfismos foram amplificados por PCR seguida de sequenciamento com primers específicos. RESULTADOS: Foram estudados 97 pacientes, com idade variando entre dois e 17 anos (7,433±4,075). Para o polimorfismo em UGT1A1, foram encontrados de cinco até oito repetições (TA) e as frequências do alelo selvagem (TA)6 e do alelo mutante (TA)7 foram respectivamente de 0,60 e 0,34. Quarenta pacientes (41,24%) eram homozigotos para o alelo (TA)6, 33 heterozigotos (TA)6/7, e 13 homozigotos (TA)7/7. Os níveis de bilirrubina total e indireta foram significativamente maiores nos pacientes com sete ou mais repetições (TA), p=0,0038 e p=0,0022, respectivamente, em comparação aos homozigotos (TA)6/6 ou heterozigotos (TA)5/6 ou (TA)5/7. Foi encontrada uma correlação negativa significativa entre os níveis de hemoglobina fetal e os níveis de bilirrubina total (r=-0,3782, p=0,0001) e de bilirrubina indireta (r=-0,3761, p=0,0015), sendo que para os indivíduos (TA)7/7 e (TA)7/8, essa correlação deixou de ser válida (p>0,05), tanto para a bilirrubina total quanto para a bilirrubina indireta. Ademais, os níveis maiores de bilirrubina nos indivíduos (TA)7/7 e (TA)7/8 não estavam associados com o aumento dos níveis de marcadores de hemólise. A frequência do alelo mutante T do rs1427407 foi de 0,22 e a do alelo selvagem G do rs7606173 foi 0,57. Nenhum indivíduo com o genótipo mutante TT para o rs1427407 estava sob tratamento com hidroxiureia. Em indivíduos sem uso de hidroxiureia, o maior nível de hemoglobina fetal foi associado com o genótipo mutante TT. Já para o genótipo selvagem GG do rs7606173 ocorreu aumento de hemoglobina fetal, mas não foi significativo. Quanto aos níveis de bilirrubina, os homozigotos TT para o rs1427407 e GG para o rs7606173 apresentaram níveis menores. Além disso, o efeito dos polimorfismos em BCL11A sobre a bilirrubina pareceu superar o efeito do rs8175347 no grupo (TA)7/7 e (TA)7/8. Não foi encontrada associação TT/CC, e também não foi encontrado nenhum mutante TT entre os portadores do haplótipo CAR/CAR. Conclusões: Este estudo confirmou a associação do alelo (TA)7 a níveis elevados de bilirrubina, independentemente do grau de hemólise. O alelo mutante T do rs1427407 e o alelo ancestral G do rs7606173 são associados à níveis maiores de hemoglobina fetal e menores níveis de bilirrubina, sendo o efeito maior associado ao alelo T. / ABSTRACT INTRODUCTION: Serum bilirubin levels are frequently increased in patients with sickle cell anemia, who already are affected by exacerbated hemolysis, which worsens symptoms and especially favors the occurrence of pigment gallstones. AIM: The aim of the present study was to analyze the influence of the polymorphism A(TA)nTAA in the UGT1A1 gene promoter (rs8175347) and two polymorphisms in the BCL11A gene (rs1427407 and rs7606173), on some laboratory parameters and on the response to hydroxyurea in pediatric patients with sickle cell anemia. MATERIAL AND METHODS: Whole blood and serum samples were used to perform hematological and biochemical analysis. DNA was extracted from peripheral blood samples, and fragments containing the polymorphisms were amplified by Polymerase Chain Reaction followed by sequencing with specific primers. RESULTS: We studied 97 patients, ranging in age from 2 to 17 years (7.433±4.075). For the UGT1A1 polymorphism, five to eight (TA) repeats were found and the wild-type (TA)6 and mutant allele (TA)7 frequencies were respectively 0.60 and 0.34. Forty patients (41.24%) were homozygous (TA)6/6, 33 heterozygotes (TA)6/7, and 13 homozygous (TA)7/7. Total and unconjugated bilirubin levels were significantly higher in patients with seven or more (TA) repeats, p=0.0038 and p=0.0022, respectively, compared to homozygotes (TA)6/6 or heterozygotes (TA)5/6 or (TA)5/7. A significant negative correlation was found between fetal hemoglobin levels and total bilirubin levels (r =-0.3782, p=0.0001) and unconjugated bilirubin levels (r =-0.3761, p=0.0015). This correlation was no longer valid (p> 0.05) for individuals (TA)7/7 and (TA)7/8. In addition, the higher levels of bilirubin in (TA)7/7 and (TA)7/8 individuals were not associated with increased markers of markers of hemolysis. The frequency of the mutant allele G of rs1427407 was 0.22, and for the wild-type allele G of rs7606173, the frequency was 0.57. In individuals that were not using hydroxyurea, a significant higher fetal hemoglobin level was associated with the TT mutant genotype, while a non-significant increase occurred for the wild-type genotype GG of rs7606173. Subsequently, patients homozygous for the TT genotype of rs1427407 had significant lower levels of total and unconjugated bilirubin (p=0.0102 and p=0.0108, respectively). Furthermore, the effect of the BCL11A polymorphisms on bilirubin levels appeared to outweigh the effect of rs8175347 in the group (TA)7/7 and (TA)7/8. No patient simultaneously carrying the homozygous mutant genotypes TT/CC was found; and no mutant homozygotes TT was found in patients with the CAR/CAR haplotype. CONCLUSION: This study confirmed that allele (TA)7 is associated with elevated levels of bilirubin, regardless of the rate of hemolysis. The mutant allele T of rs1427407 and the ancestral allele G of rs7606173 are associated with higher levels of fetal hemoglobin and lower levels of bilirubin, the strongest effect being associated with the T allele.

Class of 2007 Abstract / Objectives: The objective of this study was to determine the cost-effectiveness of using the Invader® UGT1A1 Molecular Assay to identify patients at risk of experiencing neutropenia from irinotecan monotherapy and FOLFIRI. Methods: Publicly available search engines were used to search for literature reporting the rate, treatment, and cost of adverse events and the population rate of genotypes. Drug acquisition costs were determined using average wholesale prices. Additional costs were determined using the Physicians’ Fee and Coding Guide. Monte Carlo Simulations were performed to reveal average cost, average effectiveness, and 95% confidence intervals. Tornado diagrams were derived to identify variables most likely to affect the outcomes and sensitivity analyses. Results: Monte Carlo simulations of monotherapy revealed a mean cost of treatment with genotype assay information of $6,620 (SD: $3,235; 95% CI: $5,408 to $15, 258) and an effectiveness ratio of 0.877 (SD: 0.328; 95% CI: 0 to 1). Without the genotype assay information the mean cost of treatment was $7,358 (SD: $4,183; 95% CI: $5,083 to $14,883) and the effectiveness ratio was 0.764 (SD: 0.425; 95% CI: 0 to 1). Monte Carlo simulations of FOLFIRI data showed a mean cost of treatment with genotype assay information of $6,105 (SD: $3,438; 95% CI: $4,706 to $14,556) and an effectiveness ratio of 0.858 (SD: 0.349; 95% CI: 0 to 1). When genotype information was unknown, the mean cost of FOLFIRI was $6,833 (SD: $4,288; 95% CI: $4,331 to $14,181) and the effectiveness ratio was 0.746 (SD: 0.435; 95% CI: 0 to 1). Conclusions: Genotyping patients exposed to irinotecan monotherapy or FOLFIRI may be a cost-effective means of identifying at risk patients but the results from this study while favorable are not statistically significant.

Doctor of Philosophy / Department of Chemistry / Ryan Rafferty / 6-Thiopurine (6TP), or 6-mercaptopurine, is an actively prescribed drug in the treatment of acute lymphocytic leukemia since 1952. Although 6TP has beneficial and promising therapeutic uses, severe toxicities are associated with its use such as jaundice and hepatotoxicity. These toxicities are due to the higher level of accumulation of bilirubin within the body. The bilirubin pathway involves the conjugation of water-insoluble bilirubin to two equivalents of UDPglucuronic acid (UDPGA) forming the water-soluble and excretable bilirubin diglucuronide species. The glucuronidation of bilirubin is catalyzed by the UDP-glucuronosyl transferase (UGT) enzyme, and the formation of substrate UDPGA is catalyzed by the UDP-glucose dehydrogenase enzyme (UDPGDH).The therapeutic activity of 6TP comes from two main routes: methylation of the thiol of 6TP and formation of a deoxy-6-thioguanosine triphosphate mimic that is incorporated into DNA resulting in apoptosis. In conjugation to its therapeutic metabolism, there are also detoxification pathways operating simultaneously that significantly reduce its therapeutic activity. In this pathway, oxidative metabolites of 6TP such as 8-hydroxyl-6-thiopurine (6TP-8OH), 6-thioxanthine (6TX) and 6-thiouric acid (6TU) can be formed by xanthine oxidase. It has been observed that the body retains 6TU well beyond 24-hour post 6TP treatment. Therefore, we proposed that the observed toxicity from 6TP administration comes from either 6TP or its oxidative excretion metabolites’ ability to inhibit one or both enzymes (UDPGDH, UGT) in the bilirubin excretion pathway. To investigate the toxicity resulting from the 6TP administration about these two enzymatic steps, inhibition analysis of these oxidative metabolites on UDPGDH was assessed using a robust UV-Vis method. The inhibition profile made with regards to varying UDP-glucose showed weak to no inhibition of 6TP towards UDPGDH with a K[subscript]i of 288 μΜ. However, 6TU, (the primary oxidative metabolite which is oxidized at C2 and C8) has increased inhibition towards UDPGDH with K[subscript]i of 7 μΜ. Inhibition was also observed with 6TX (oxidized at C2) and 8-OH-6TP (oxidized at C8) with K[subscript]i values 54 and 14 μΜ, respectively. To further confirm the results of the UV-Vis assessment, inhibition studies were carried out using an HPLC method that was developed and validated to separate all the analytes in the UDPGDH catalyzed reaction. Inhibition studies were performed via the HPLC method showed K[subscript]i values of 105 μΜ and 5 μΜ for 6TP and 6TU, respectively, towards UDPGDH. To assess the inhibition studies towards the UGT enzyme, an HPLC method was developed for the simultaneous determination of bilirubin and its mono/diglucuronides. The inhibition studies were carried to assess the formation of glucuronides and consumption of UDPGA in the presence of the inhibitors using the HPLC method developed. Neither 6TP nor 6TU were shown to inhibit UGT. Also, inhibition studies were carried out in vivo animal model, which further confirmed that 6TP and 6TU do inhibit UDPGDH, but no effect on UGT activity. With these results, we discovered that both 6TP and its oxidative metabolites inhibit UDPGDH. Furthermore, it was observed that C2 and C8 positions of 6TP are important for the toxicity towards UDPGDH. With the goal of developing a single multi-enzymatic assay, another HPLC method was developed to assess the UDPGDH and UGT catalyze reactions together. This method can be used as a standard method to assess interference of any molecule on bilirubin excretion. Given the findings in this study, efforts are being directed towards the synthesis of 8-substituted 6TP analogs that are proposed to retain its therapeutic efficacy but have limited to no toxicity.

Kiekvieno vaisto sukeltas tiek farmakologinis (veiksmingumo), tiek toksikologinis poveikis kiekvienam pacientui yra skirtingas, todėl gana dažnai vaistų skyrimas ir vartojimas tampa labai komplikuotas. Prancūzijoje 3.2% hospitalizacijos atvejų yra sąlygoti vaistų. Tai sudaro 320 milijonų eurų sumą per metus. Genetiniai faktoriai, sąlygojantys vaistų farmakokinetiką ir farmakodinamiką, dalinai paaiškina skirtingą vaistų poveikį žmogui. Tyrimo tikslas: 1. Patikrinti galimybę gausinti serume ir plazmoje esančią DNR nauju būdu ir atlikti genetinius tyrimus. 2. Patvirtinti realaus laiko polimerazės grandininės reakcijos (PGR) metodiką aromatazės genui, panaudojus ląsteles, kurių šio geno raiška yra pakankama. Rezultatai: Patikrinta galimybė gausinti serume ir plazmoje esančią DNR, sekvenavus gautą DNR ir sekas palyginus su esančiomis duomenų bazėje : rezultatai teigiami. Nustatyta, kad, ekstrahuojant druskiniu metodu, gaunama daugiau DNR, bet ji blogesnės kokybės, nei naudojant QIAGEN kolonėles. Patvirtinta realaus laiko PGR metodika aromatazės genui, naudojant krūties vėžio ląsteles. / There is much variability in the manner individuals respond to drugs, such that the management of some drugs is problematic. In France, the incidence of hospital admissions related to adverse drug reactions is estimated to be 3.2 %, at an annual cost of over 300 millions euros. Genetic factors affecting the pharmacokinetics and pharmacodynamics of drugs partly explain interindividual variability in drug response. Aim of experiment: 1. verify, if it is possible to amplify serum and plasma DNA using new method, and make a genetic research. 2. verify real time polymerase chain reaction to aromatase gene. Find cell line, whish have a sufficient expression of this gene. Results : We verified the possibility to amplify serum and plasma DNA using new method. We made the sequencing of DNA extracts and we compared results in data base : results are positives. It`s important, that QIAGEN extracts are cleanner than salt extracts, but there are few of DNA. We confirmed real time PCR method to aromatase gene, using breast cancer cells.

Inherited hyperbilirubinemias are a group of metabolic disorders, characterized by increased levels of total serum bilirubin or its conjugated fraction. Most of these hyperbilirubinemias are inherited autosomal recessively and are manifested in young age. Increased bilirubin reflects the genetic disturbances in one of the enzymes of heme degradation pathway, the defect of bilirubin conjugation (UGT1A1 gene) or its transport (ABCC2, OATP1B1, OATP1B3). All of these proteins are involved not only in elimination of bilirubin, but various substrates; therefore the performed studies have a great pharmacogenomics impact. We have studied the molecular pathology of hereditary hyperbilirubinemias in Caucasian and Roma population and to compare the clinical and biochemical results with the molecular genetic data. We described the impact of compound defect of c.-3279T>G and g.175492_175493insTA on total serum bilirubin and calculated the linkage disequlibrium of these two variants in promoter region of UGT1A1 gene. We also verified, that the population distribution of both variants is in concordance with the literature. In our second study, we have described the rare conjugated hyperbilirubinemia Dubin-Johnson type among 7 Roma families. We have found a novel variant NG_011798.1:c.[1013_1014delTG] together with...

Inherited hyperbilirubinemias are a group of metabolic disorders, characterized by increased levels of total serum bilirubin or its conjugated fraction. Most of these hyperbilirubinemias are inherited autosomal recessively and are manifested in young age. Increased bilirubin reflects the genetic disturbances in one of the enzymes of heme degradation pathway, the defect of bilirubin conjugation (UGT1A1 gene) or its transport (ABCC2, OATP1B1, OATP1B3). All of these proteins are involved not only in elimination of bilirubin, but various substrates; therefore the performed studies have a great pharmacogenomics impact. We have studied the molecular pathology of hereditary hyperbilirubinemias in Caucasian and Roma population and to compare the clinical and biochemical results with the molecular genetic data. We described the impact of compound defect of c.-3279T>G and g.175492_175493insTA on total serum bilirubin and calculated the linkage disequlibrium of these two variants in promoter region of UGT1A1 gene. We also verified, that the population distribution of both variants is in concordance with the literature. In our second study, we have described the rare conjugated hyperbilirubinemia Dubin-Johnson type among 7 Roma families. We have found a novel variant NG_011798.1:c.[1013_1014delTG] together with...

A icterícia costuma ser um fenômeno fisiológico em recém-nascidos saudáveis, contudo, hiperbilirrubinemia não conjugada grave ocorre em 5 a 6% dessa população. Tem sido sugerido que a variabilidade genética de genes de proteínas transportadoras de bilirrubina poderia aumentar o risco de hiperbilirrubinemia quando associada a outros fatores icterogênicos. O presente estudo incluiu neonatos com idade gestacional de 35 semanas ou mais e peso superior a 2000g com indicação para fototerapia. Os polimorfismos nos genes SLCO1B1 (rs4149056 e rs2306283) e SLCO1B3 (rs17680137 e rs2117032) foram analisados por sondas de hidrólise. Um total de 167 hiperbilirrubinêmicos e 247 recém-nascidos controles foi arrolado. Sexo, incompatibilidade ABO, peso ao nascimento e idade gestacional diferiram entre os grupos, mas as frequências alélicas e genotípicas dos polimorfismos dos genes SLCO1B não. Na análise por regressão logística, incompatibilidade ABO, idade gestacional e o alelo polimórfico T do rs2117032 permaneceram no modelo. A presença desse polimorfismo pareceu conferir proteção à hiperbilirrubinemia. Os indivíduos que eram homozigotos para o alelo G do rs2306283 e deficientes da enzima glicose-6-fosfato desidrogenase eram mais frequentes entre os casos. No presente estudo foram observadas apenas fracas interações entre os diferentes polimorfismos e os fatores clínico-ambientais. / Jaundice usually is a physiological phenomenon among newborns; however, severe hyperbilirubinemia occurs in 5% to 6% of this population. It has been suggested that genetic variation in the genes of bilirubin transporters could enhance the risk of hyperbilirubinemia when co-expressed with other icterogenic conditions. The present study included newborns with a gestational age of greater than 35 weeks and weights greater than 2000 g with indications for phototherapy. The polymorphisms from SLCO1B1 (rs4149056 and rs2306283) and SLCO1B3 (rs17680137 and rs2117032) genes were analyzed by hydrolysis probes. A total of 167 hyperbilirubinemic infants and 247 control subjects were enrolled. The gender, ABO incompatibility, birth weight, and gestational age differed between the groups but the allelic and genotypic frequency of the polymorphisms from SLCO1B genes did not. In logistic regression, the ABO incompatibility, gestational age, and polymorphic T allele of rs2117032 remained in the model. The presence of this polymorphism seemed to provide protection from hyperbilirubinemia. The individuals that were homozygous for the G allele of rs2306283 and glucose 6-phosphate-dehydrogenase deficient were more frequent among the cases. In the present study there were observed only weak associations between the different polymorphisms and the clinical/environmental factors.

A icterícia costuma ser um fenômeno fisiológico em recém-nascidos saudáveis, contudo, hiperbilirrubinemia não conjugada grave ocorre em 5 a 6% dessa população. Tem sido sugerido que a variabilidade genética de genes de proteínas transportadoras de bilirrubina poderia aumentar o risco de hiperbilirrubinemia quando associada a outros fatores icterogênicos. O presente estudo incluiu neonatos com idade gestacional de 35 semanas ou mais e peso superior a 2000g com indicação para fototerapia. Os polimorfismos nos genes SLCO1B1 (rs4149056 e rs2306283) e SLCO1B3 (rs17680137 e rs2117032) foram analisados por sondas de hidrólise. Um total de 167 hiperbilirrubinêmicos e 247 recém-nascidos controles foi arrolado. Sexo, incompatibilidade ABO, peso ao nascimento e idade gestacional diferiram entre os grupos, mas as frequências alélicas e genotípicas dos polimorfismos dos genes SLCO1B não. Na análise por regressão logística, incompatibilidade ABO, idade gestacional e o alelo polimórfico T do rs2117032 permaneceram no modelo. A presença desse polimorfismo pareceu conferir proteção à hiperbilirrubinemia. Os indivíduos que eram homozigotos para o alelo G do rs2306283 e deficientes da enzima glicose-6-fosfato desidrogenase eram mais frequentes entre os casos. No presente estudo foram observadas apenas fracas interações entre os diferentes polimorfismos e os fatores clínico-ambientais. / Jaundice usually is a physiological phenomenon among newborns; however, severe hyperbilirubinemia occurs in 5% to 6% of this population. It has been suggested that genetic variation in the genes of bilirubin transporters could enhance the risk of hyperbilirubinemia when co-expressed with other icterogenic conditions. The present study included newborns with a gestational age of greater than 35 weeks and weights greater than 2000 g with indications for phototherapy. The polymorphisms from SLCO1B1 (rs4149056 and rs2306283) and SLCO1B3 (rs17680137 and rs2117032) genes were analyzed by hydrolysis probes. A total of 167 hyperbilirubinemic infants and 247 control subjects were enrolled. The gender, ABO incompatibility, birth weight, and gestational age differed between the groups but the allelic and genotypic frequency of the polymorphisms from SLCO1B genes did not. In logistic regression, the ABO incompatibility, gestational age, and polymorphic T allele of rs2117032 remained in the model. The presence of this polymorphism seemed to provide protection from hyperbilirubinemia. The individuals that were homozygous for the G allele of rs2306283 and glucose 6-phosphate-dehydrogenase deficient were more frequent among the cases. In the present study there were observed only weak associations between the different polymorphisms and the clinical/environmental factors.

A icterícia costuma ser um fenômeno fisiológico em recém-nascidos saudáveis, contudo, hiperbilirrubinemia não conjugada grave ocorre em 5 a 6% dessa população. Tem sido sugerido que a variabilidade genética de genes de proteínas transportadoras de bilirrubina poderia aumentar o risco de hiperbilirrubinemia quando associada a outros fatores icterogênicos. O presente estudo incluiu neonatos com idade gestacional de 35 semanas ou mais e peso superior a 2000g com indicação para fototerapia. Os polimorfismos nos genes SLCO1B1 (rs4149056 e rs2306283) e SLCO1B3 (rs17680137 e rs2117032) foram analisados por sondas de hidrólise. Um total de 167 hiperbilirrubinêmicos e 247 recém-nascidos controles foi arrolado. Sexo, incompatibilidade ABO, peso ao nascimento e idade gestacional diferiram entre os grupos, mas as frequências alélicas e genotípicas dos polimorfismos dos genes SLCO1B não. Na análise por regressão logística, incompatibilidade ABO, idade gestacional e o alelo polimórfico T do rs2117032 permaneceram no modelo. A presença desse polimorfismo pareceu conferir proteção à hiperbilirrubinemia. Os indivíduos que eram homozigotos para o alelo G do rs2306283 e deficientes da enzima glicose-6-fosfato desidrogenase eram mais frequentes entre os casos. No presente estudo foram observadas apenas fracas interações entre os diferentes polimorfismos e os fatores clínico-ambientais. / Jaundice usually is a physiological phenomenon among newborns; however, severe hyperbilirubinemia occurs in 5% to 6% of this population. It has been suggested that genetic variation in the genes of bilirubin transporters could enhance the risk of hyperbilirubinemia when co-expressed with other icterogenic conditions. The present study included newborns with a gestational age of greater than 35 weeks and weights greater than 2000 g with indications for phototherapy. The polymorphisms from SLCO1B1 (rs4149056 and rs2306283) and SLCO1B3 (rs17680137 and rs2117032) genes were analyzed by hydrolysis probes. A total of 167 hyperbilirubinemic infants and 247 control subjects were enrolled. The gender, ABO incompatibility, birth weight, and gestational age differed between the groups but the allelic and genotypic frequency of the polymorphisms from SLCO1B genes did not. In logistic regression, the ABO incompatibility, gestational age, and polymorphic T allele of rs2117032 remained in the model. The presence of this polymorphism seemed to provide protection from hyperbilirubinemia. The individuals that were homozygous for the G allele of rs2306283 and glucose 6-phosphate-dehydrogenase deficient were more frequent among the cases. In the present study there were observed only weak associations between the different polymorphisms and the clinical/environmental factors.

Bilirubin is a major product of the heme catabolism in the vascular bed with substantial antioxidant properties. These importantly contribute to pathogenesis of diseases associated with increased oxidative stress, including cardiovascular or cancer diseases. In the first part of this PhD project serum bilirubin concentrations were examined in the 1 % representative sample of the general Czech population, together with determination of the prevalence of Gilbert's syndrome. Bilirubin concentrations were determined also within individual polymorphisms of the UGT1A1 gene (OMIM*191740) responsible for bilirubin biotransformation in the liver, including their association with the basic risk factors for atherosclerosis. We also assessed the activity of the standard liver enzymes (representing another significant risk factor for the development of cardiovascular diseases) with surprisingly high proportion of subjects with elevated values. Simultaneously, we determined the concentrations of serum bilirubin in a group of patients with an acute coronary syndrome, who manifested with significantly lower concentrations as compared to general population. In the second part of this research project, the relationship between plasma concentrations of bilirubin and individual variants of UGT1A1 gene polymorphisms...