RESUMO
BACKGROUND: Increased vitamin B6 catabolism related to inflammation, as measured by the PAr index (the ratio of 4-pyridoxic acid over the sum of pyridoxal and pyridoxal-5'-phosphate), has been positively associated with lung cancer risk in two prospective European studies. However, the extent to which this association translates to more diverse populations is not known. MATERIALS AND METHODS: For this study, we included 5323 incident lung cancer cases and 5323 controls individually matched by age, sex, and smoking status within each of 20 prospective cohorts from the Lung Cancer Cohort Consortium. Cohort-specific odds ratios (ORs) and 95% confidence intervals (CIs) for the association between PAr and lung cancer risk were calculated using conditional logistic regression and pooled using random-effects models. RESULTS: PAr was positively associated with lung cancer risk in a dose-response fashion. Comparing the fourth versus first quartiles of PAr resulted in an OR of 1.38 (95% CI: 1.19-1.59) for overall lung cancer risk. The association between PAr and lung cancer risk was most prominent in former smokers (OR: 1.69, 95% CI: 1.36-2.10), men (OR: 1.60, 95% CI: 1.28-2.00), and for cancers diagnosed within 3 years of blood draw (OR: 1.73, 95% CI: 1.34-2.23). CONCLUSION: Based on pre-diagnostic data from 20 cohorts across 4 continents, this study confirms that increased vitamin B6 catabolism related to inflammation and immune activation is associated with a higher risk of developing lung cancer. Moreover, PAr may be a pre-diagnostic marker of lung cancer rather than a causal factor.
Assuntos
Inflamação/sangue , Neoplasias Pulmonares/sangue , Metabolismo , Vitamina B 6/sangue , Adulto , Idoso , Feminino , Humanos , Inflamação/patologia , Neoplasias Pulmonares/epidemiologia , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Ácido Piridóxico/metabolismo , Fatores de Risco , FumantesRESUMO
BACKGROUND: Cigarette smoking has been identified as a major modifiable risk factor for coronary heart disease and mortality. However, findings on the relationship between smoking and atrial fibrillation (AF) have been inconsistent. Furthermore, findings from previous studies were based on self-reported smoking. OBJECTIVE: To examine the associations of smoking status and plasma cotinine levels, a marker of nicotine exposure, with risk of incident AF in the Hordaland Health Study. METHODS: We conducted a prospective analysis of 6682 adults aged 46-74 years without known AF at baseline. Participants were followed via linkage to the Cardiovascular Disease in Norway (CVDNOR) project and the Cause of Death Registry. Smoking status was assessed by both questionnaire and plasma cotinine levels. RESULTS: A total of 538 participants developed AF over a median follow-up period of 11 years. Using questionnaire data, current smoking (HR: 1.41, 95% CI: 1.09-1.83), but not former smoking (HR: 1.03, 95% CI: 0.83-1.28), was associated with an increased risk of AF after adjustment for gender, age, body mass index, hypertension, physical activity and education. Using plasma cotinine only, the adjusted HR (95% CI) was 1.40 (1.12-1.75) for participants with cotinine ≥85 nmol L-1 compared to those with cotinine <85 nmol L-1 . However, the risk increased with elevated plasma cotinine levels until 1199 nmol L-1 (HR: 1.55, 95% CI: 1.16-2.05 at the third group vs. the reference group) and plateaued at higher levels. CONCLUSIONS: Current, but not former smokers, had a higher risk of developing AF. Use of plasma cotinine measurement corroborated this finding.
Assuntos
Fibrilação Atrial , Fumar Cigarros , Cotinina/sangue , Idoso , Fibrilação Atrial/sangue , Fibrilação Atrial/diagnóstico , Fibrilação Atrial/epidemiologia , Biomarcadores/sangue , Fumar Cigarros/epidemiologia , Fumar Cigarros/metabolismo , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Nicotina/metabolismo , Noruega/epidemiologia , Estudos Prospectivos , Fatores de RiscoRESUMO
BACKGROUND: Disturbances in one carbon metabolism may contribute to carcinogenesis by affecting methylation and synthesis of DNA. Choline and its oxidation product betaine are involved in this metabolism and can serve as alternative methyl group donors when folate status is low. PATIENTS AND METHODS: We conducted a case-control study nested within the European Prospective Investigation into Cancer and Nutrition (EPIC), to investigate plasma concentrations of the methyl donors methionine, choline, betaine (trimethylglycine), and dimethylglycine (DMG) in relation to colorectal cancer (CRC) risk. Our study included 1367 incident CRC cases (965 colon and 402 rectum) and 2323 controls matched by gender, age group, and study center. Multivariate-adjusted odds ratios (ORs) and 95% confidence intervals (95% CIs) for CRC risk were estimated by conditional logistic regression, comparing the fifth to the first quintile of plasma concentrations. RESULTS: Overall, methionine (OR: 0.79, 95% CI: 0.63-0.99, P-trend = 0.05), choline (OR: 0.77, 95% CI: 0.60-0.99, P-trend = 0.07), and betaine (OR: 0.85, 95% CI: 0.66-1.09, P-trend = 0.06) concentrations were inversely associated with CRC risk of borderline significance. In participants with folate concentration below the median of 11.3 nmol/l, high betaine concentration was associated with reduced CRC risk (OR: 0.71, 95% CI: 0.50-1.00, P-trend = 0.02), which was not observed for those having a higher folate status. Among women, but not men, high choline concentration was associated with decreased CRC risk (OR: 0.62, 95% CI: 0.43-0.88, P-trend = 0.01). Plasma DMG was not associated with CRC risk. CONCLUSIONS: Individuals with high plasma concentrations of methionine, choline, and betaine may be at reduced risk of CRC.
Assuntos
Betaína/sangue , Colina/sangue , Neoplasias Colorretais/etiologia , Metionina/sangue , Estado Nutricional/fisiologia , Sarcosina/análogos & derivados , Idoso , Estudos de Casos e Controles , Neoplasias Colorretais/sangue , Neoplasias Colorretais/epidemiologia , Europa (Continente)/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Fatores de Risco , Sarcosina/sangueRESUMO
The risk of osteoporosis increases in inflammatory disorders. In cell-mediated immune activation, interferon (IFN)-γ stimulates macrophage release of neopterin and increases the activity of indoleamine 2,3-dioxygenase (IDO), thereby stimulating tryptophan degradation along the kynurenine pathway. Plasma levels of neopterin and the kynurenine/tryptophan ratio (KTR) are thus markers of IFN-γ-mediated inflammation. Several kynurenine pathway metabolites (kynurenines) possess immunomodulatory properties. The aim of this study was to investigate associations between markers of IFN-γ-mediated inflammation and kynurenines with bone mineral density (BMD). The community-based Hordaland Health Study (HUSK), with middle-aged (46-49 years) and older (71-74 years) participants, was conducted from 1998 to 2000 (n = 5312). Hip BMD in relation to neopterin, KTR and kynurenines were investigated, using linear and logistic regression analyses. In the oldest group, neopterin (P ≤ 0·019) and KTR (P ≤ 0·001) were associated inversely with BMD after multiple adjustment. Comparing the highest to the lowest quartiles, the odds ratios of low BMD (being in the lowest quintile of BMD) in the oldest cohort were for neopterin 2·01 among men and 2·34 among women (P ≤ 0·007) and for KTR 1·80 for men and 2·04 for women (P ≤ 0·022). Xanthurenic acid was associated positively with BMD in all sex and age groups while 3-hydroxyanthranilic acid was associated positively with BMD among women only (P ≤ 0·010). In conclusion, we found an inverse association between BMD and markers of IFN-γ-mediated inflammation in the oldest participants. BMD was also associated with two kynurenines in both age groups. These results may support a role of cell-mediated inflammation in bone metabolism.
Assuntos
Densidade Óssea , Inflamação/metabolismo , Interferon gama/metabolismo , Cinurenina/metabolismo , Redes e Vias Metabólicas , Idoso , Biomarcadores/metabolismo , Feminino , Humanos , Inflamação/sangue , Mediadores da Inflamação/sangue , Mediadores da Inflamação/metabolismo , Cinurenina/sangue , Masculino , Metaboloma , Pessoa de Meia-Idade , Neopterina/sangue , Neopterina/metabolismo , Fatores de Risco , Triptofano/sangueRESUMO
Circulating neopterin and kynurenine/tryptophan ratio (KTR) increase during inflammation and serve as markers of cellular immune activation, but data are sparse on other determinants of these markers and metabolites of the kynurenine pathway. We measured neopterin, tryptophan, kynurenine, anthranilic acid, kynurenic acid, 3-hydroxykynurenine, 3-hydroxyanthranilic acid and xanthurenic acid in plasma in two age groups, 45-46 years (n = 3723) and 70-72 years (n = 3329). Differences across categories of the potential determinants, including age, gender, renal function, body mass index (BMI), smoking and physical activity, were tested by Mann-Whitney U-test and multiple linear regression including age group, gender, renal function and lifestyle factors. In this multivariate model, neopterin, KTR and most kynurenines were 20-30% higher in the older group, whereas tryptophan was 7% lower. Men had 6-19% higher concentrations of tryptophan and most kynurenines than women of the same age. Compared to the fourth age-specific estimated glomerular filtration rate (eGFR) quartile, the first quartile was associated with higher concentrations of neopterin (25%) and KTR (24%) and 18-36% higher concentrations of kynurenines, except 3-hydroxyanthranilic acid. Additionally, KTR, tryptophan and all kynurenines, except anthranilic acid, were 2-8% higher in overweight and 3-17% higher in obese, than in normal-weight individuals. Heavy smokers had 4-14% lower levels of tryptophan and most kynurenines than non-smokers. Age and renal function were the strongest determinants of plasma neopterin, KTR and most kynurenines. These findings are relevant for the design and interpretation of studies investigating the role of plasma neopterin, KTR and kynurenines in chronic diseases.
Assuntos
Envelhecimento/imunologia , Inflamação/sangue , Ácido 3-Hidroxiantranílico/análise , Idoso , Envelhecimento/sangue , Biomarcadores/sangue , Índice de Massa Corporal , Creatinina/sangue , Feminino , Humanos , Ácido Cinurênico/sangue , Cinurenina/análogos & derivados , Cinurenina/sangue , Masculino , Pessoa de Meia-Idade , Atividade Motora , Neopterina/sangue , Noruega , Valores de Referência , Fumar/sangue , Triptofano/sangue , Xanturenatos/sangue , ortoaminobenzoatos/sangueRESUMO
BACKGROUND: The aim of the present study was to assess potential long-term reduction in health-related quality of life (HRQOL) in adult trauma patients 2-7 years after discharge from an intensive care unit (ICU), and to study possible determinants of the HRQOL reduction. METHODS: Follow-up study of a cohort of 341 trauma patients admitted to the ICU of a university hospital during 1998-2003. Of the 228 eligible patients, 210 (92%) completed the study. A telephone interview using the EuroQol 5-D (EQ-5D) was conducted. Patients reported their HRQOL both at present and before trauma. RESULTS: Before trauma 88% reported in retrospect no problem in any EQ-5D dimension, compared with 20% at follow-up. After trauma (median 4.0 years) 58% suffered pain/discomfort, 44% reported alterations in usual activities, 40% reduced mobility, 35% anxiety/depression, and 15% limited autonomy. A total of 74% experienced reduction in HRQOL. Severe problems were reported by 16%. Women experienced more anxiety/depression than men. Simplified Acute Physiology Score (SAPS) II and Injury Severity Score (ISS) were significantly associated with impaired HRQOL, while age was not. Patients with severe head injury reported better HRQOL than those without severe head injury. CONCLUSION: More than 2 years post-injury, 74% reported impaired HRQOL but only 16% had severe problems. The majority still suffered pain/discomfort, indicating that pain management is a key factor in improving long-term outcome after severe trauma.
Assuntos
Cuidados Críticos/psicologia , Unidades de Terapia Intensiva/estatística & dados numéricos , Avaliação de Resultados em Cuidados de Saúde/estatística & dados numéricos , Qualidade de Vida , Perfil de Impacto da Doença , Ferimentos e Lesões/psicologia , Atividades Cotidianas , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Estudos de Coortes , Cuidados Críticos/estatística & dados numéricos , Feminino , Seguimentos , Humanos , Entrevistas como Assunto , Masculino , Transtornos Mentais/epidemiologia , Transtornos Mentais/psicologia , Pessoa de Meia-Idade , Limitação da Mobilidade , Noruega/epidemiologia , Dor/epidemiologia , Dor/psicologia , Qualidade de Vida/psicologia , Anos de Vida Ajustados por Qualidade de Vida , Fatores Sexuais , Fatores de Tempo , Ferimentos e Lesões/epidemiologiaRESUMO
BACKGROUND: In India, most people adhere to a vegetarian diet, which may lead to cobalamin deficiency. OBJECTIVE: The objective was to examine indicators of cobalamin status in Asian Indians. DESIGN: The study population included 204 men and women aged 27-55 y from Pune, Maharashtra, India, categorized into 4 groups: patients with cardiovascular disease (CVD) and diabetes, patients with CVD but no diabetes, patients with diabetes but no CVD, and healthy subjects. Data on medical history, lifestyle, and diet were obtained by interviews and questionnaires. Blood samples were collected for measurement of serum or plasma total cobalamin, holotranscobalamin (holoTC), methylmalonic acid (MMA), and total homocysteine (tHcy) and hemetologic indexes. RESULTS: MMA, tHcy, total cobalamin, and holoTC did not differ significantly among the 4 groups; therefore, the data were pooled. Total cobalamin showed a strong inverse correlation with tHcy (r = -0.59) and MMA (r = -0.54). Forty-seven percent of the subjects had cobalamin deficiency (total cobalamin <150 pmol/L), 73% had low holoTC (<35 pmol/L), 77% had hyperhomocysteinemia (tHcy >15 micromol/L), and 73% had elevated serum MMA (>0.26 micromol/L). These indicators of impaired cobalamin status were observed in both vegetarians and nonvegetarians. Folate deficiency was rare and only 2.5% of the subjects were homozygous for the MTHFR 677C-->T polymorphism. CONCLUSIONS: About 75% of the subjects had metabolic signs of cobalamin deficiency, which was only partly explained by the vegetarian diet. If impaired cobalamin status is confirmed in other parts of India, it may have important health implications.
Assuntos
Homocisteína/sangue , Hiper-Homocisteinemia/sangue , Ácido Metilmalônico/sangue , Deficiência de Vitamina B 12/diagnóstico , Vitamina B 12/sangue , Adulto , Biomarcadores , Doenças Cardiovasculares/sangue , Doenças Cardiovasculares/fisiopatologia , Diabetes Mellitus/sangue , Diabetes Mellitus/fisiopatologia , Dieta Vegetariana , Feminino , Ácido Fólico/sangue , Humanos , Índia/epidemiologia , Masculino , Pessoa de Meia-Idade , Prevalência , Inquéritos e Questionários , Deficiência de Vitamina B 12/sangue , Deficiência de Vitamina B 12/epidemiologiaRESUMO
Most studies demonstrate increased risk of colorectal cancer (CRC) and adenomas in folate-deficient subjects or that high folate intake may afford some protection. Smoking increases such risk in some but not all studies. We investigated whether smoking, folate status and methylenetetrahydrofolate reductase (MTHFR) genotype predict the risk of adenomatous and hyperplastic polyps of colorectum. By colonoscopy, the type, number, size and extent of dysplasia of colorectal polyps were assessed in 443 subjects aged 63-72 years. We also determined RBC folate and the C667T polymorphism of the methylenetetrahydrofolate reductase (MTHFR) gene. Smoking, folate status and the C677T MTHFR polymorphism were strong, interactive determinants of high-risk adenomas (HRAs, defined as adenomas > or =10 mm in diameter, adenomas with villous components or with severe dysplasia). The risk was particularly high in smokers with low folate and the CT/TT genotype (risk category T) and in smokers with high folate and the CC genotype (risk category C). With non-smokers with low folate and the CC genotype as reference, the odds ratios (OR, 95% CI) were 8.7 (2.5-29.7) in category T and 9.9 (2.6-38.4) in category C. Notably, this risk pattern was also observed for hyperplastic polyps. In conclusion, in smokers, high folate status may confer increased or decreased risk for HRAs, depending on the MTHFR genotype. These data demonstrate the strong gene-nutrition interaction involving the C677T MTHFR polymorphism.
Assuntos
Pólipos Adenomatosos/patologia , Neoplasias Colorretais/patologia , Ácido Fólico/sangue , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/genética , Fumar/efeitos adversos , Pólipos Adenomatosos/etiologia , Pólipos Adenomatosos/genética , Idoso , Neoplasias Colorretais/etiologia , Neoplasias Colorretais/genética , Estudos Transversais , Feminino , Frequência do Gene , Genótipo , Homocisteína/sangue , Humanos , Hiperplasia , Masculino , Metilenotetra-Hidrofolato Redutase (NADPH2) , Pessoa de Meia-Idade , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/sangue , Fatores de RiscoRESUMO
BACKGROUND: The aims of this cohort study were to assess the survival of trauma patients treated in a general intensive care unit (ICU) and to evaluate the simplified acute physiology score (SAPS) II, maximum sequential organ failure assessment (SOFA) score, injury severity score (ISS), age, sex and severe head injury as predictors of 30-day mortality. METHODS: Three hundred and twenty-five adult patients admitted during 1998-2003 were evaluated retrospectively with update of survival data in January 2005. Kaplan-Meier statistics and Cox proportional hazards regression were used to study survival and to assess predictors of mortality, respectively. RESULTS: The 30-day mortality was 16.9%, ICU mortality 13.8% and hospital mortality 17.8%. Long-term survival (observation time, 1-7 years) was 77.8%. After 3.5 years, mortality was the same as for the background population. Severe head injury was the main cause of death and increased the risk of 30-day mortality 2.4-fold. In addition, SAPS II and an age above 50 years proved to be significant predictors of mortality in a multivariate analysis. Sex was not associated with mortality, and ISS and the maximum SOFA score were significant predictors in univariate analyses only. CONCLUSION: Reduced long-term survival was observed up to 3.5 years after acute injury. The 30-day mortality was strongly related to severe head injury, SAPS II and an age above 50 years. These variables may be useful as predictors of mortality, and may contribute to risk adjustment of this subset of trauma patients when treatment results from different centres are compared.
Assuntos
Mortalidade Hospitalar , Ferimentos e Lesões/mortalidade , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Traumatismos Craniocerebrais/mortalidade , Métodos Epidemiológicos , Feminino , Humanos , Unidades de Terapia Intensiva , Masculino , Pessoa de Meia-Idade , Noruega/epidemiologia , Sistema de Registros , Projetos de Pesquisa , Fatores de TempoRESUMO
Published studies and our own preliminary results show that Intensive Care Unit (ICU) patients admitted after severe trauma, all report reduced quality of life (QOL) after discharge. In contrast to other ICU patients, this reduction is decreased by time, but does not seem to reach pre-ICU levels. Interventions to improve this situation have not been documented. This should be a high priority aspect for all health personnel caring for patients surviving severe trauma.
Assuntos
Cuidados Críticos , Qualidade de Vida , Ferimentos e Lesões/terapia , HumanosRESUMO
BACKGROUND: Successful thoracic epidural analgesia depends on the sensory blockage of specific dermatomes following appropriate placement of the epidural catheter. This study aimed to ascertain how accurately anaesthesiologists identify thoracic intervertebral spaces, and whether counting from the prominent vertebra is easier than using the iliac crest as an anatomical landmark. METHODS: Five anaesthesiologists attempted to locate one out of five consecutive intervertebral spaces (Th7-Th8 to Th11-Th12) on patients referred for magnetic resonance imaging of the vertebral column. The intended thoracic interspace and the counting reference point (C7-Th1 or L3-L4) were marked with oil capsules. The body mass index, gender and position of the patient were recorded. The exact capsule positions were determined by a radiologist after the study. RESULTS: In 92 patients, 26.7% of the thoracic interspaces were correctly identified. The counting reference point was the only variable studied with a significant influence on error. The accuracy increased when the iliac crest was used as an anatomical landmark rather than the prominent vertebra (odds ratio, 0.29). The majority (76.4%) of all the incorrectly placed capsules were found cephalad to the intended level. CONCLUSION: We recommend that the caudal of two to three possible interspaces should be used when placing an epidural catheter in the thoracic spine. Because of the inaccurate localization of the thoracic intervertebral spaces, documentation should state the site of puncture as being in the upper or lower thoracic spine instead of claiming to be in an exact interspace.
Assuntos
Anestesia Epidural , Disco Intervertebral/anatomia & histologia , Vértebras Torácicas/anatomia & histologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Índice de Massa Corporal , Feminino , Humanos , Modelos Logísticos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-IdadeRESUMO
Since 1984 children treated in the Department of Burns, Haukeland Hospital, have been evaluated by a child psychiatrist. This article describes the psychological reactions of the children and their parents, and the psychiatric support and treatment provided during the stay in hospital.
Assuntos
Queimaduras/psicologia , Queimaduras/terapia , Criança , Psiquiatria Infantil , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , PsicoterapiaRESUMO
The psychological consequences of trauma on 28 Swedish children were examined 1 year and 3 years after a bus accident in western Norway in 1988. Symptoms were assessed by the Impact of Event Scale (IES), semi-structured interviews, and a questionnaire developed for this study. At the 1-year follow-up, large proportions of the sample had symptoms of Intrusion and Avoidance (IES) and symptoms of general psychological distress, but no clinically significant symptoms were observed at the 3-year follow-up. Girls and children who experienced loss in the accident were characterised by high levels of Intrusion, whereas passenger trauma and age were unrelated to the outcome. A bidirectional relationship was observed between the mothers' and the children's symptoms over time, whereas the fathers' symptoms were unrelated to the children's symptoms. The clinical implications of the study are that symptoms in all children of traumatised families deserve attention during the first year post-trauma, and that objective risk indicators (age, sex, and types of trauma) do not provide sufficient information to identify children at risk.
Assuntos
Acidentes de Trânsito/psicologia , Veículos Automotores , Transtornos de Estresse Pós-Traumáticos/psicologia , Adaptação Psicológica , Adolescente , Criança , Transtornos Reativos da Criança/diagnóstico , Transtornos Reativos da Criança/psicologia , Feminino , Seguimentos , Pesar , Humanos , Masculino , Traumatismo Múltiplo/psicologia , Noruega , Inventário de Personalidade , Transtornos de Estresse Pós-Traumáticos/diagnóstico , Suécia/etnologiaRESUMO
In August 1988 a Swedish schoolbus with 23 children and 11 adult passengers crashed into a tunnel wall in Norway. Twelve children and four parents died, 18 of the passengers survived. The victims' relatives (parents, spouses, siblings, and others) were transported to Norway and given a "confronting" support program at the University hospital in Bergen. The crisis intervention program is described: how the relatives were helped to confront and actively cope with some of the stressful situations from which traumatized families very often are protected. One year later 42 of the relatives answered a questionnaire evaluating the confronting support program that was offered during their stay in Norway. A majority of the respondents did not regret their participation in the program. The answers indicate that passivity and helplessness can be counteracted through a combination of confrontation with reality and emotional support.
Assuntos
Acidentes de Trânsito/psicologia , Intervenção em Crise , Teste de Realidade , Sobrevida/psicologia , Adaptação Psicológica , Adolescente , Adulto , Idoso , Luto , Criança , Família/psicologia , Feminino , Humanos , Controle Interno-Externo , Masculino , Pessoa de Meia-Idade , Traumatismo Múltiplo/psicologia , Noruega , Apoio Social , Transtornos de Estresse Pós-Traumáticos/psicologiaRESUMO
In capillary electrophoresis (CE), separation of DNA fragments is usually performed in covalently coated capillaries. Recent studies have demonstrated that certain polymers form a dynamic coating on the inner surface of the capillary, thereby suppressing the electroosmotic flow and DNA-capillary wall interactions. We developed a simple method for the synthesis of short-chain polydimethylacrylamide (PDMA) using isopropanol as a chain transfer agent. Capillary (<75 microm internal diameter) filling and replacement of this low-viscosity (14 cP at 4% PDMA) self-coating medium were easily carried out by commercial CE instruments. Using PDMA and uncoated capillaries, we first examined the separation of phi X174 HaeIII DNA digests and observed that the stability of the dynamic coating was markedly better at pH 7.8 than at pH 8.3. At this lower pH and nondenaturing conditions, high resolution of the phi X174 HaeIII DNA digests was obtained for more than 850 injections in the same capillary. We then exploited this sieving medium in CE using multiple approaches for mutation analysis of clinical DNA samples including separation of restriction enzyme cleavage products, analysis of single strand conformation polymorphisms, and simultaneous detection of several mutations using multiplex allele-specific PCR amplification. Our results demonstrate that CE in uncoated capillaries using PDMA as sieving medium is a simple, versatile, and reliable strategy for separation and mutation analysis of clinical DNA samples.
Assuntos
Resinas Acrílicas , Análise Mutacional de DNA/métodos , DNA/genética , DNA/isolamento & purificação , Eletroforese Capilar/métodos , Resinas Acrílicas/síntese química , Resinas Acrílicas/química , Bacteriófago phi X 174/química , Sequência de Bases , Primers do DNA/genética , DNA Viral/isolamento & purificação , Humanos , Concentração de Íons de Hidrogênio , Polimorfismo Conformacional de Fita SimplesRESUMO
The prognosis for pancreatic carcinoma is generally poor. The chance of survival could be improved if curative surgery were performed, but early diagnosis is then essential. Fine-needle aspiration cytology and endoscopic retrograde cholepancreaticography (ERCP) are widely used in order to obtain a precise diagnosis before laparotomy. In almost all patients with pancreatic adenocarcinomas, the oncogene Ki-ras is activated by point mutation. We describe a patient where conventional diagnostic procedures were inconclusive. However, DNA-analysis of aspirate from the pancreatic duct and from a pancreatic cyst showed activated Ki-ras oncogene. A malignant diagnosis was later confirmed. We propose that DNA-analyses of pancreatic fluid or tissues should be used whenever facing problems with an early and accurate diagnosis of pancreatic lesions.
Assuntos
Adenocarcinoma/diagnóstico , Genes ras , Suco Pancreático/química , Neoplasias Pancreáticas/diagnóstico , Adenocarcinoma/genética , Adenocarcinoma/mortalidade , Idoso , Análise Mutacional de DNA , DNA de Neoplasias/análise , Feminino , Humanos , Pâncreas/diagnóstico por imagem , Pâncreas/patologia , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/mortalidade , Prognóstico , Tomografia Computadorizada por Raios XRESUMO
A rapid analysis of single-strand conformation polymorphism (SSCP) by capillary electrophoresis with laser-induced fluorescence (CE-LIF) detector was developed using a short-chain, linear polyacrylamide (PA) as sieving medium. Capillary filling of this low-viscosity medium and medium replacement were carried out by commercial capillary electrophoresis instruments. The approach was successfully applied to detect the C677T mutation of methylenetetrahydrofolate reductase gene. The influences of factors such as the concentration of polymers, voltage, temperature, and additives on the SSCP analysis were systematically investigated. Using 6% PA sieving medium and high electric field, four strands were resolved within 11 min in a DNA sample heterozygous for the C677T mutation, and a characteristic pattern was apparent for each of the three genotypes. When using multiple injection mode, the average analysis time per sample was reduced to about 4 min. In conclusion, our results indicate that CE-LIF may be an alternative to conventional SSCP analysis based on slab gel electrophoresis for the detection of genetic mutations. The technique is simple and rapid and is well suited to analysis of large numbers of clinical samples.
Assuntos
Resinas Acrílicas , Eletroforese Capilar/métodos , Polimorfismo Conformacional de Fita Simples , DNA/química , Humanos , Lasers , Reação em Cadeia da Polimerase , Espectrometria de FluorescênciaRESUMO
We report on a patient who suffered an acute, extensive intracerebral haemorrhage, leading to symptoms of cerebral herniation within a few hours. The clinical diagnosis of brain death was made based on a neurological examination, and an apnoea test eight hours after the haemorrhage. A few hours later the diagnosis was changed, as several reflexes reappeared. After six days mechanical ventilation was withdrawn, as the brain damage was considered so serious as to render further therapy futile. It was considered unethical to sustain therapy for a possible organ donation at a later date. A review of relevant the literature, however, shows that brain-dead patients may exhibit such varying degrees of autonomic and spinal reflexes as to cause confusion, thus delaying the physician in making a diagnosis. Often, an opportunity for organ donation is lost. Based on this review, we believe that our patient was indeed brain dead when the first diagnosis was made, and that a cerebral angiography should have been performed. Because organ donation is an important issue, the diagnosis of brain death must be definitive.
Assuntos
Morte Encefálica , Reflexo , Dano Encefálico Crônico/diagnóstico , Dano Encefálico Crônico/fisiopatologia , Morte Encefálica/diagnóstico , Hemorragia Cerebral/diagnóstico , Hemorragia Cerebral/fisiopatologia , Feminino , Humanos , Pessoa de Meia-Idade , Exame Neurológico , Ordens quanto à Conduta (Ética Médica)RESUMO
The C677T mutation of the methylenetetrahydrofolate reductase gene and the G1691A (Leiden) mutation of the factor V gene are established risk factors for thromboembolic disease. We here present an assay for the simultaneous genotyping of these common genetic variants. The assay involves a strategy based on multiplex mutagenically separated PCR performed in a single tube containing six primers. Separation of the resulting four PCR products (197, 207, 233, and 246 bp) was performed by capillary electrophoresis coupled with laser-induced fluorescence detection. The time for the automated electrophoresis was reduced to 2.5 min per sample by performing the capillary electrophoresis analysis in a multiple-injection mode.