A phylogenetic study of eleven cetaceans was undertaken using Restriction Endonuclease Maps (RSM) of mitochondrial DNA (mtDNA). One species from the suborder mysticeti (baleen whales) was sampled, and of the ten odontocetes (toothed whales) sampled two were from the family Ziphiidae (beaked whales) and eight were from the family Delphinidae (dolphins) (each representing a different genus). The primarily opportunistically obtained (i.e. from strandings or accidental death in commercial trawl nets) heart tissue generally yielded high quantities of mtDNA which is needed for double digest fragment analysis. The mtDNA extracted from the sampled taxa was cleaved with fifteen different six-base Restriction Enzymes (RE's). Using the three-way method of analysis and aided by the computer program Resolve (Ver. 2.7) (Harley, unpublished), RSM's were constructed. Distance (Neighbor-Joining and Fitsch-Margoliash) and cladistic (Maximum Parsimony and Bootstrap) methods were used to infer phylogenies. The baleen whale was used as an outgroup for the cladistic analysis. Both the distance and both the cladistic methods produced the same single topology, which is concordant with morphologically based classifications. The two differences (within the Delphinidae), viz. Grampus' most basally rooted position and Cephalorhynchus' grouping with the Delphininae are of taxa whose groupings are unresolved in the morphologically based classifications. Using Brown et al's (1979) molecular clock, very recent divergence times at the generic, family and suborder levels were obtained, when compared to fossil based estimates. Using the odontoceti/mysticeti split the base substitution rate of cetacean mtDNA was estimated to be much slower than that of terrestrial mammals (0,3% compared to 1,0% Myr⁻¹). A similarly slow rate was calculated for cetacean nuclear DNA (nDNA) (0,09% Myr⁻¹) (Schlotterer et al, 1991). It remains an unresolved issue as to whether the base substitution rate of cetacean DNA is slower than terrestrial mammals or whether the fossil evidence needs to be reinterpreted. The time of the mysticeti/odontoceti split is palaeontologically uncertain and the suggested monophyletic status of the extant suborders has been questioned, thus making the calculation of cetacean base substitution rate risky. Equally, the incomplete fossil record can lend itself to misinterpretation.

Cheung Shiu-fai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 131-139). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABSTRACT --- p.iii / 摘要 --- p.vi / TABLE OF CONTENTS --- p.ix / LIST OF FIGURES --- p.xvi / LIST OF TABLES --- p.xviii / ABBREVIATIONS --- p.xix / PUBLICATION --- p.xxi / AWARD --- p.xix / Chapter SECTION 1. --- INTRODUCTION OF HEPATOCELLULAR CARCINOMA --- p.1 / Chapter 1.1 --- Epidemiology of Hepatocellular Carcinoma --- p.1 / Chapter 1.2 --- Etiologies of HCC --- p.1 / Chapter 1.2.1 --- Hepatitis B Virus and Hepatitis C Virus --- p.2 / Chapter 1.2.2 --- Aflatoxins and Alcohol --- p.3 / Chapter 1.3 --- Major Diagnostic and Prognostic Markers of HCC --- p.4 / Chapter 1.3.1 --- Biochemical Tumor Markers --- p.5 / Chapter 1.3.2 --- Clinico-pathological Features of HCC --- p.6 / Chapter SECTION 2. --- THE MITOCHONDRION --- p.7 / Chapter 2.1 --- Structure of the Mitochondrial Genome --- p.9 / Chapter 2.1.1 --- Nicotinamide Adenine Dinucleotide Dehydrogenase --- p.10 / Chapter 2.1.2 --- Cytochrome b --- p.10 / Chapter 2.1.3 --- Cytochrome c Oxidase --- p.11 / Chapter 2.1.4 --- ATP Synthase --- p.11 / Chapter 2.1.5 --- Ribosomal RNA --- p.11 / Chapter 2.1.6 --- Transfer RNA --- p.12 / Chapter 2.1.7 --- Displacement Loop --- p.12 / Chapter 2.2 --- Replication of Mitochondrial DNA --- p.17 / Chapter 2.3 --- Transcription of Mitochondrial DNA --- p.17 / Chapter SECTION 3. --- PHYSIOLOGY OF MITOCHONDRIA --- p.19 / Chapter 3.1 --- Energy Production by Oxidative Phosphorylation (OXPHOS) --- p.19 / Chapter 3.2 --- Programmed Cell Death: Apoptosis --- p.22 / Chapter 3.3 --- Morphology of Mitochondria in Hepatocytes --- p.25 / Chapter SECTION 4. --- MUTATIONS OF MITOCHONDRIAL DNA --- p.26 / Chapter 4.1 --- Special Terms Used in This Study --- p.26 / Chapter 4.1.1 --- Somatic Mutations and Polymorphisms --- p.26 / Chapter 4.1.2 --- Homoplasmic and Heteroplasmic Mutations --- p.26 / Chapter 4.2 --- Factors Causing High Mutation Frequency in mtDNA --- p.27 / Chapter 4.2.1 --- Presence of Reactive Oxygen Species --- p.27 / Chapter 4.2.2 --- Lack of Protective Histories --- p.28 / Chapter 4.2.3 --- Limited DNA Repair Mechanism --- p.29 / Chapter 4.3 --- Theories of Homoplasmic Mutations --- p.31 / Chapter 4.3.1 --- Replicative Advantage on Mutated mtDNA Sequence Selection --- p.31 / Chapter 4.3.2 --- Random Mutagenesis and Segregation --- p.32 / Chapter 4.4 --- MtDNA Mutations in Mitochondrial Disease and Aging --- p.33 / Chapter 4.5 --- MtDNA Deletions in Cancer --- p.34 / Chapter 4.6 --- Somatic Mutations of MtDNA in Various Cancers --- p.35 / Chapter 4.6.1 --- Frequencies of Somatic Mutations --- p.35 / Chapter 4.6.2 --- Distribution of Somatic Mutations in mtDNA --- p.36 / Chapter 4.7 --- Somatic Mutations of Mitochondrial DNA in HCC --- p.37 / Chapter SECTION 5. --- OBJECTIVES OF THIS STUDY --- p.44 / Chapter SECTION 6. --- MATERIALS AND METHODS --- p.46 / Chapter 6.1 --- Patients and Samples Collection --- p.46 / Chapter 6.2 --- DNA Extraction from Liver Tissues --- p.46 / Chapter 6.3 --- Amplification of Mitochondrial DNA by Polymerase Chain Reaction --- p.51 / Chapter 6.3.1 --- Design of Primers --- p.51 / Chapter 6.3.2 --- PCR Conditions and Contents --- p.54 / Chapter 6.3.3 --- Assessment of PCR Products by Agarose Gel Electrophoresis --- p.54 / Chapter 6.4 --- Purification of PCR Products --- p.54 / Chapter 6.5 --- Cyclesequencing of Mitochondrial DNA --- p.55 / Chapter 6.5.1 --- Design of Primers --- p.55 / Chapter 6.5.2 --- PCR Contents and Cycle Sequencing Procedures --- p.56 / Chapter 6.6 --- Purification of Sequencing Products --- p.56 / Chapter 6.7 --- Sequence Analysis by Automated Sequencer --- p.57 / Chapter 6.7.1 --- Preparation of Polyacrylamide Gel --- p.57 / Chapter 6.7.2 --- Sequence Analysis by Automated Sequencer --- p.58 / Chapter 6.7.3 --- "Search for Sequence Variants, Polymorphisms and Somatic Mutations" --- p.58 / Chapter 6.8 --- Further Studies on mtDNA Mutations --- p.59 / Chapter 6.8.1 --- Sequence Analysis in Buffy Coat --- p.59 / Chapter 6.8.2 --- Detection of the Presence of Somatic mtDNA Mutations in Plasma --- p.59 / Chapter 6.8.3 --- Frequency of Mutations in Two Nucleotide Repeat Sequences --- p.60 / Chapter 6.9 --- Clinical Data and Statistical Analysis --- p.61 / Chapter 6.9.1 --- Clinical and Pathological Data --- p.61 / Chapter 6.9.2 --- Statistical Analysis --- p.61 / Chapter SECTION 7. --- RESULTS --- p.63 / Chapter 7.1 --- Sequence Analysis of the Entire Mitochondrial Genome --- p.63 / Chapter 7.1.1 --- Sequence Variants and Polymorphisms --- p.63 / Chapter 7.1.2 --- Somatic Mutations --- p.71 / Chapter 7.2 --- Study of Mitochondrial Sequence in Lymphocytes --- p.78 / Chapter 7.3 --- Detection of Tumor DNA in Serum --- p.78 / Chapter 7.4 --- Analysis of Nucleotide Repeat Sequences --- p.79 / Chapter 7.4.1 --- General Results --- p.79 / Chapter 7.4.2 --- Statistical Analysis --- p.84 / Chapter SECTION 8. --- DISCUSSION --- p.89 / Chapter 8.1 --- Comparative Analysis of mtDNA Mutations with Two Previous HCC Studies --- p.89 / Chapter 8.1.1 --- Number of Cases and Region Studied --- p.89 / Chapter 8.1.2 --- Number and Distribution of Mutations in Normal Controls --- p.89 / Chapter 8.1.3 --- Number of Somatic Mutations --- p.90 / Chapter 8.1.4 --- Distribution of Somatic Mutations --- p.91 / Chapter 8.2 --- Similarities of Somatic mtDNA Mutations in This Study with Other Cancer Types --- p.93 / Chapter 8.2.1 --- Frequency and Distribution of Somatic Mutations --- p.93 / Chapter 8.2.2 --- Number of Homoplasmic Mutations --- p.93 / Chapter 8.3 --- Evaluation of Somatic Mutations of mtDNA in This Study --- p.96 / Chapter 8.3.1 --- Specificity of Somatic Mutations in Tumor Proved by Sequence Analysis in Lymphocytes --- p.96 / Chapter 8.3.2 --- Importance of Conserved Amino Acid Sequences with Other Species to the Presence of Somatic Mutations in Tumor --- p.96 / Chapter 8.3.3 --- Four Somatic Mutation Sites Are Detected in More than One Cancer Type --- p.101 / Chapter 8.3.4 --- Presence of Homoplasmic and Heteroplasmic Mutations --- p.101 / Chapter 8.3.5 --- Absence of Large-scale Deletions in Tumor Tissues --- p.102 / Chapter 8.4 --- Mutation Hotspots Region: Hypervariable Displacement-loop --- p.103 / Chapter 8.5 --- D310 Mononucleotide Repeats --- p.106 / Chapter 8.5.1 --- Description of D310 Mononucleotide Repeats --- p.106 / Chapter 8.5.2 --- Possible Causes of Varied Sequences at D310 --- p.106 / Chapter 8.5.3 --- Appearance of Nucleotide Repeats at D310 in Tumors --- p.107 / Chapter 8.5.4 --- Possible Outcomes of D310 Aberrations in mtDNA Replication and Transcription --- p.108 / Chapter 8.5.5 --- Comparison of D310 Alternations in HCC with Other Cancers --- p.109 / Chapter 8.6 --- Other Nucleotide Repeat Sequences --- p.112 / Chapter 8.6.1 --- The CA Dinucleotide Repeats --- p.112 / Chapter 8.6.2 --- Other Nucleotide Repeat Sequences Showing Genome Instability --- p.112 / Chapter 8.7 --- Evaluation of Somatic mtDNA Mutations as a Cancer Diagnostic Marker --- p.114 / Chapter 8.7.1 --- Coding Region --- p.114 / Chapter 8.7.2 --- D-loop Region --- p.114 / Chapter 8.7.3 --- D310 Nucleotide Repeats --- p.115 / Chapter 8.7.4 --- Possibility of Detecting Somatic Mutations in Serum --- p.116 / Chapter 8.8 --- Somatic mtDNA Mutations May Be a Prognostic Marker in HCC --- p.117 / Chapter 8.8.1 --- Possible Problems in Current Prognostic Factors --- p.117 / Chapter 8.8.2 --- Interpretation of Results --- p.117 / Chapter 8.8.3 --- Prognostic Values of Somatic Mutations at D310 --- p.118 / Chapter 8.9 --- Hypothesis of Somatic MtDNA Mutations on Tumorigenesis and Tumor Progression --- p.119 / Chapter 8.9.1 --- Somatic mtDNA Mutations Decline OXPHOS and May Inactivate Apoptotic Pathways --- p.119 / Chapter 8.9.2 --- Moderate Reactive Oxygen Species Production May Promote Mitosis --- p.120 / Chapter 8.10 --- Possible Appearance of Somatic Mutations in HCC with Chronic HBV Infection --- p.123 / Chapter 8.11 --- Possibility of HBx Protein Integration to MtDNA Mutations --- p.123 / Chapter 8.12 --- Conclusions --- p.125 / Chapter SECTION 9. --- LIMITATIONS AND FURTHER STUDIES --- p.127 / Chapter 9.1 --- Limitations and Improvements of Study --- p.127 / Chapter 9.1.1 --- Small Sample Size --- p.127 / Chapter 9.1.2 --- Sequence Analysis Method --- p.127 / Chapter 9.1.3 --- Fidelity of PCR Reactions and Long-range PCR Fragments --- p.128 / Chapter 9.2 --- Further Studies --- p.129 / Chapter SECTION 10. --- REFERENCES --- p.131

A espécie Aedes scapularis é um dos culicídeos de grande importância médica. Está distribuída nas Américas e tem grande competência vetora para diversos arbovírus. No estado de São Paulo, há ocorrência de Ae. scapularis em vários municípios, como em Pariquera-Açu e São Paulo. O objetivo desse trabalho foi testar se há diferenciação genética - morfológica entre essas populações, podendo diagnosticar existência de espécies crípticas em Aedes scapularis. As populações estudadas foram Pariquera-Açu (PAR), Parque Ecológico do Tietê em São Paulo (PET) e Butantã (BUT). Os parâmetros utilizados foram: Morfometria geométrica da asa (forma e tamanho), estudo do gene mitocondrial COI, análise dos espaçadores internos transcritos ITS2 e análise morfológica de ovos. Com os resultados obtidos, podemos concluir que a divergência populacional é atestada por padrões geográficos de forma alar, gene mitocondrial COI e razão comprimento e largura dos ovos, mas extensões de estudos em outras áreas precisam ser feitos para poder atestar espécies crípticas em Aedes scapularis. / The species Aedes scapularis is a culicidae of medical importance. It is distributed in the Americas and has a high vector competence for many arboviruses. In state of São Paulo, have occurrence of Ae. scapularis in many cities, such as Pariquera-Acu and the city of São Paulo. The aim of this study was to determine differentiation genetic- morphology among these populations, being able to diagnose the existence of cryptic species in Aedes scapularis. The populations studied were Pariquera-Acu (PAR), the Tietê Ecological Park in Sao Paulo (PET) and Butantã (BUT). The parameters used were: wing geometric morphometry (shape and size), study of mitochondrial gene COI, analysis of internal transcribed spacers ITS2 and morphological analysis of eggs. With these results, we conclude that divergence population is attested by the geographical patterns of wing shape, and COI mitochondrial gene length and width ratio of eggs, but extensions of studies in other areas need to be made in order to attest cryptic species in Aedes scapularis.

Thesis (M.Sc. (Tropical Medicine))--Mahidol University, 2003.

Orientadores: Nadja Cristhina de Souza Pinto, Anibal Eugenio Vercesi / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-24T01:38:55Z (GMT). No. of bitstreams: 1 Costa_RuteAlvesPereirae_D.pdf: 10770990 bytes, checksum: 08f31895722a5f60be02fcde15d3ea05 (MD5) Previous issue date: 2013 / Resumo: Xeroderma Pigmentosum (XP) é uma doença rara, autossômica recessiva, caracterizada por fotossensibilidade, mudanças pigmentares, envelhecimento precoce da pele e incidência elevada de neoplasias de pele. XP é causada por mutações em, pelo menos oito genes, que caracterizam sete diferentes grupos de complementação genética (XP-A a XP-G) e um tipo variante (XP-V). Mutações em cada em dos genes envolvidos resultam em diferentes graus de severidade da doença, principalmente quanto ao comprometimento neurológico. Pacientes XP-C apresentam mutações no gene Xpc, que resultam, geralmente, em proteínas truncadas e instáveis. XPC é uma proteína envolvida na via de reparo de DNA por excisão de nucleotídeos (NER) e sua função é reconhecer a lesão na fita de DNA e dar início ao reparo. Recentemente, a participação indireta de XPC no reparo por excisão de bases (BER) foi sugerida, através de sua interação física e funcional com a DNA glicosilase OGG1. Uma vez que OGG1 é essencial para a remoção de purinas oxidadas do DNA mitocondrial, nós hipotetizamos que o DNAmt, e consequentemente a função mitocondrial, estariam comprometidas em células deficientes em XPC. Desta forma, este trabalho se propôs a investigar alterações bioenergéticas mitocondrias em células obtidas de pacientes XP-C. Nossos resultados revelaram que linhagens celulares XP-C apresentavam menor função mitocondrial, apesar de não apresentarem alterações no número de cópias de DNAmt. O consumo de oxigênio pelo complexo I estava significativamente diminuído em células XP-C quando comparado à células controle, enquanto que o consumo de O2 via os complexos II, III e IV foi maior em células XP-C. A capacidade de captar cálcio também se mostrou alterada nas células XP-C, uma vez que essa célula era incapaz de captar e reter concentrações fisiológicas desse íon. A produção de espécies reativas de oxigênio foi significativamente maior em células XP-C comparadas a células controle. Em acordo, a atividade das enzimas antioxidantes superóxido dismutase e glutationa peroxidase foi menor em células XP-C, indicando um desbalanço redox nessas células. A análise da expressão de genes relacionados à biogênese mitocondrial revelou que um regulador transcricional fundamental, o coativador PGC1?, estava significativamente reduzido em células XP-C transformadas e primárias. Resultados de Western blotting e imunofluorescência revelaram que as alterações bioenergéticas e genômicas observadas em células XP-C eram via sinalização e não por efeito direto, uma vez que nas condições experimentais utilizadas neste trabalho, XPC não está presente na mitocôndria. Nossos resultados demonstram, pela primeira vez, que a proteína XPC exerce um papel indireto na manutenção da integridade funcional da mitocôndria, provavelmente através de seu papel no controle da expressão de genes envolvidos na biogênese mitocondrial / Abstract: Xeroderma pigmentosum (XP) is a rare autosomal recessive disorder characterized by photosensitivity, pigmentary changes, premature skin aging and increased incidence of skin cancer. XP is caused by mutations in at least eight genes, which characterize seven different genetic complementation groups (XP-A to XP-G) and variant type (XP-V). Mutations in each gene result in varying degrees of severity, mostly regarding the presence or not of neurodegeneration. XP-C is caused by mutations in the Xpc gene, resulting, mostly, in a truncated and unstable protein. The XPC protein is involved in the nucleotide excision repair pathway (NER), where it functions as a damage recognition factor. Recently, a role for XPC in the base excision repair (BER) pathway has been proposed, through its physical and fucntional interaction with the DNA glycosylase OGG1. Since OGG1 has a major function in repairing oxidized purines in the mitochondrial DNA (mtDNA), we hypothesized that XPC played a role in maitaining mtDNA integrity, and consequently, mitochondrial function. Thus, this study proposes to investigate mitocondrial function in XP-C cell. Our results showed that XP-C cells had less mitochondrial function, although without changes in mtDNA copy number. Oxygen consumption through complex I was lower in XP-C cells compared to control cells, while respiration through complexes II, III and IV was higher in XP-C cells. Calcium uptake and retention by mitochondria was also decreased in XP-C cells, as these cells were unable to retain even physiological spikes in calcium concentration. Reactive oxygen species production was significantly higher in XPC cells compared to controls. In agreement to that, the activity of the antioxidant enzymes superoxide dismutase and glutathione peroxidase was significatly decreased in XP-C cells, indicating that these cells are under a severe redox signaling inbalance. The analysis of the expression of genes related to mitochondrial biogenesis revealed that the key transcriptional regulator PGC1? was significantly lower in both transformed and primary XP-C cells. The results of Western blotting and imunofluorescence revealed that the bioenergetic impairment observed in XP-C cells is likely the result of changes in expression and signaling pathwyas, since, under the experimental conditions used here, XPC is not present in mitochondria. Our results indicate, for the first time, that XPC plays an important role in mitochondrial maintenace, likely via its role in transcription regulation of mitochondrial biogenesis / Doutorado / Fisiopatologia Médica / Doutora em Ciências

A espécie Aedes scapularis é um dos culicídeos de grande importância médica. Está distribuída nas Américas e tem grande competência vetora para diversos arbovírus. No estado de São Paulo, há ocorrência de Ae. scapularis em vários municípios, como em Pariquera-Açu e São Paulo. O objetivo desse trabalho foi testar se há diferenciação genética - morfológica entre essas populações, podendo diagnosticar existência de espécies crípticas em Aedes scapularis. As populações estudadas foram Pariquera-Açu (PAR), Parque Ecológico do Tietê em São Paulo (PET) e Butantã (BUT). Os parâmetros utilizados foram: Morfometria geométrica da asa (forma e tamanho), estudo do gene mitocondrial COI, análise dos espaçadores internos transcritos ITS2 e análise morfológica de ovos. Com os resultados obtidos, podemos concluir que a divergência populacional é atestada por padrões geográficos de forma alar, gene mitocondrial COI e razão comprimento e largura dos ovos, mas extensões de estudos em outras áreas precisam ser feitos para poder atestar espécies crípticas em Aedes scapularis. / The species Aedes scapularis is a culicidae of medical importance. It is distributed in the Americas and has a high vector competence for many arboviruses. In state of São Paulo, have occurrence of Ae. scapularis in many cities, such as Pariquera-Acu and the city of São Paulo. The aim of this study was to determine differentiation genetic- morphology among these populations, being able to diagnose the existence of cryptic species in Aedes scapularis. The populations studied were Pariquera-Acu (PAR), the Tietê Ecological Park in Sao Paulo (PET) and Butantã (BUT). The parameters used were: wing geometric morphometry (shape and size), study of mitochondrial gene COI, analysis of internal transcribed spacers ITS2 and morphological analysis of eggs. With these results, we conclude that divergence population is attested by the geographical patterns of wing shape, and COI mitochondrial gene length and width ratio of eggs, but extensions of studies in other areas need to be made in order to attest cryptic species in Aedes scapularis.

Introdução: A endometriose afeta 10 a 15% das mulheres em idade reprodutiva e há evidências crescentes de que o estresse oxidativo está envolvido na sua patogênese. Os polimorfismos na região controladora do DNA mitocondrial podem levar à replicação e à transcrição alterada dos genes mitocondriais, o que pode afetar a função mitocondrial e, consequentemente, a geração intracelular de espécies reativas de oxigênio. Descobertas recentes indicam que os polimorfismos do DNA mitocondrial (mtDNA) estão relacionados à endometriose em populações coreana e indiana. Objetivo: Avaliar a associação entre polimorfismos e haplogrupos do DNA mitocondrial e a presença de endometriose em mulheres brasileiras. Métodos: Pacientes com idade entre 18 e 50 anos foram divididas nos grupos endometriose (n = 90) e controle (n = 92). O primeiro grupo incluiu mulheres com diagnóstico histológico de endometriose e estadiamento cirúrgico, enquanto o segundo grupo incluiu pacientes submetidas à cirurgia para laqueadura tubária, leiomioma ou cistos ovarianos benignos, sem evidência de endometriose. O DNA foi extraído a partir de amostras de sangue periférico seguido do sequenciamento de Sanger e eletroforese capilar. Os polimorfismos foram determinados comparando as sequências obtidas com a Sequência padrão de Cambridge Revisada. Resultados: As frequências dos polimorfismos T16217C (14,4% vs. 5,4%; p = 0,049), G499A (13,3% vs. 4,3%; p = 0,038), T236C (5,6% vs. 0%; p = 0,028) e G185A (6,7% vs. 0%; p = 0.013) foram maior no grupo endometriose em comparação ao grupo controle, respectivamente, enquanto as frequências dos polimorfismos T146C (18,9% vs 32,6%; p = 0,042) e 573.2C (5,6% vs. 29,3%; p < 0.001) foram menores. A distribuição dos haplogrupos foi semelhante entre os grupos endometriose e controle. Nenhuma diferença foi observada nos haplogrupos de acordo com o estádio ou localização da doença. Conclusão: Os polimorfismos T16217C, G499A, T236C e G185A do DNA mitocondrial foram relacionados à presença de endometriose, enquanto T146C e 573.2C foram relacionados à ausência de doença, em amostra de mulheres brasileiras. Não foram observadas diferenças significativas entre os haplogrupos mitocondriais / Background: There is increasing evidence that oxidative stress is a major factor in the pathogenesis of endometriosis, a prevalent disease that affects 5-15% of reproductive-aged women worldwide. Polymorphisms in the control region of mitochondrial DNA (mtDNA) can lead to the altered replication and transcription of mitochondrial genes, thereby affecting both overall mitochondrial function, and the intracellular generation of reactive oxygen species. Objective: The present study investigated whether the incidence of mtDNA polymorphisms and/or haplogroups is associated with endometriosis in a Brazilian population. Methods: Female patients (aged 18-50 years) were enrolled in the present study, and assigned to either endometriosis (n = 90) or control (n = 92) group. The former group comprised patients who had received a histological diagnosis of endometriosis and had been assigned a surgical stage, while the latter comprised patients who had undergone gynecological surgery for tubal ligation, leiomyoma, or ovarian cysts, and showed no evidence of endometriosis. DNA was extracted from peripheral blood samples, and then subjected to Sanger sequencing and capillary electrophoresis. Polymorphisms were identified by comparing the isolated mtDNA sequences with the revised Cambridge Reference Sequence. Results: The frequency of some identified polymorphisms was found to be higher in the endometriosis group than in the control group, including polymorphisms T16217C (found in 14.4% and 5.4% of endometriosis- and control-group members, respectively; p=0.049), G499A (13.3% vs. 4.3%; p=0.038), T236C (5.6% vs. 0%; p=0.028), and G185A (6.7% vs. 0%; p=0.013). In contrast, polymorphisms T146C (18.9% vs. 32.6%; p=0.042) and 573.2C (5.6% vs. 29.3%; p < 0.001) were found to occur at a lower frequency in the endometriosis compared to the control group. Observed haplogroup frequencies were similar between the endometriosis and control groups, and did not appear to be affected by either disease location and/or staging. Conclusion: mtDNA polymorphisms T16217C, G499A, T236C, and G185A were found to be associated with endometriosis, while conversely, T146C and 573.2C were shown to be associated with an absence of disease in the analyzed Brazilian population. No significant differences were observed between the mitochondrial haplogroups of patients with, versus without endometriosis

Introdução: A endometriose afeta 10 a 15% das mulheres em idade reprodutiva e há evidências crescentes de que o estresse oxidativo está envolvido na sua patogênese. Os polimorfismos na região controladora do DNA mitocondrial podem levar à replicação e à transcrição alterada dos genes mitocondriais, o que pode afetar a função mitocondrial e, consequentemente, a geração intracelular de espécies reativas de oxigênio. Descobertas recentes indicam que os polimorfismos do DNA mitocondrial (mtDNA) estão relacionados à endometriose em populações coreana e indiana. Objetivo: Avaliar a associação entre polimorfismos e haplogrupos do DNA mitocondrial e a presença de endometriose em mulheres brasileiras. Métodos: Pacientes com idade entre 18 e 50 anos foram divididas nos grupos endometriose (n = 90) e controle (n = 92). O primeiro grupo incluiu mulheres com diagnóstico histológico de endometriose e estadiamento cirúrgico, enquanto o segundo grupo incluiu pacientes submetidas à cirurgia para laqueadura tubária, leiomioma ou cistos ovarianos benignos, sem evidência de endometriose. O DNA foi extraído a partir de amostras de sangue periférico seguido do sequenciamento de Sanger e eletroforese capilar. Os polimorfismos foram determinados comparando as sequências obtidas com a Sequência padrão de Cambridge Revisada. Resultados: As frequências dos polimorfismos T16217C (14,4% vs. 5,4%; p = 0,049), G499A (13,3% vs. 4,3%; p = 0,038), T236C (5,6% vs. 0%; p = 0,028) e G185A (6,7% vs. 0%; p = 0.013) foram maior no grupo endometriose em comparação ao grupo controle, respectivamente, enquanto as frequências dos polimorfismos T146C (18,9% vs 32,6%; p = 0,042) e 573.2C (5,6% vs. 29,3%; p < 0.001) foram menores. A distribuição dos haplogrupos foi semelhante entre os grupos endometriose e controle. Nenhuma diferença foi observada nos haplogrupos de acordo com o estádio ou localização da doença. Conclusão: Os polimorfismos T16217C, G499A, T236C e G185A do DNA mitocondrial foram relacionados à presença de endometriose, enquanto T146C e 573.2C foram relacionados à ausência de doença, em amostra de mulheres brasileiras. Não foram observadas diferenças significativas entre os haplogrupos mitocondriais / Background: There is increasing evidence that oxidative stress is a major factor in the pathogenesis of endometriosis, a prevalent disease that affects 5-15% of reproductive-aged women worldwide. Polymorphisms in the control region of mitochondrial DNA (mtDNA) can lead to the altered replication and transcription of mitochondrial genes, thereby affecting both overall mitochondrial function, and the intracellular generation of reactive oxygen species. Objective: The present study investigated whether the incidence of mtDNA polymorphisms and/or haplogroups is associated with endometriosis in a Brazilian population. Methods: Female patients (aged 18-50 years) were enrolled in the present study, and assigned to either endometriosis (n = 90) or control (n = 92) group. The former group comprised patients who had received a histological diagnosis of endometriosis and had been assigned a surgical stage, while the latter comprised patients who had undergone gynecological surgery for tubal ligation, leiomyoma, or ovarian cysts, and showed no evidence of endometriosis. DNA was extracted from peripheral blood samples, and then subjected to Sanger sequencing and capillary electrophoresis. Polymorphisms were identified by comparing the isolated mtDNA sequences with the revised Cambridge Reference Sequence. Results: The frequency of some identified polymorphisms was found to be higher in the endometriosis group than in the control group, including polymorphisms T16217C (found in 14.4% and 5.4% of endometriosis- and control-group members, respectively; p=0.049), G499A (13.3% vs. 4.3%; p=0.038), T236C (5.6% vs. 0%; p=0.028), and G185A (6.7% vs. 0%; p=0.013). In contrast, polymorphisms T146C (18.9% vs. 32.6%; p=0.042) and 573.2C (5.6% vs. 29.3%; p < 0.001) were found to occur at a lower frequency in the endometriosis compared to the control group. Observed haplogroup frequencies were similar between the endometriosis and control groups, and did not appear to be affected by either disease location and/or staging. Conclusion: mtDNA polymorphisms T16217C, G499A, T236C, and G185A were found to be associated with endometriosis, while conversely, T146C and 573.2C were shown to be associated with an absence of disease in the analyzed Brazilian population. No significant differences were observed between the mitochondrial haplogroups of patients with, versus without endometriosis

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2007. / Härtill 5 uppsatser.

Dissertation (Ph.D.)--University of South Florida, 2009. / Includes vita. Includes bibliographical references.