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Angiotensin-Converting Enzyme Gene Polymorphisms and Tissue Oxygenation During Exercise: ACE genotypes

Angiotensin-Converting Enzyme Gene Polymorphisms and Tissue Oxygenation During Exercise: ACE genotypesIn addition, a plastic catheter was placed percutaneously into the brachial artery to monitor systemic arterial pressure and to sample systemic arterial blood. Heart rate and rhythm were monitored continuously. PAP was measured using a transducer (UK901; Baxter; Tokyo, Japan) located at the level of the anterior fourth intercostal space, with the patient sitting upright, and was recorded on photographic paper. Pressures were averaged over three respiratory cycles. Mean pressures were obtained by electronic integration. Cardiac output (Qt) was determined by the thermodilution method, using a cardiac output computer (Fukuda Denshi; Tokyo, Japan). Arterial blood gas tensions were measured with a blood gas analyzer (model IL 1312; Instrumentation Laboratory; Tokyo, Japan), and the blood was rapidly deproteinated in iced perchlorate solution and, after centrifugation, was analyzed for lactate concentration by an enzymatic technique. tadanafil
Resting hemodynamic and blood gas data were obtained about 20 min after the patient had been seated comfortably on the ergometer. Each patient then underwent a constant-load exercise test for 5 min on the ergometer at a workload corresponding to 60% of the previously determined maximal workload. Hemodynamic and blood gas measurements were performed during the final minute of constant-load exercise.
Genomic DNA was extracted from peripheral blood leukocytes by standard methods. The ACE genotypes of the subjects were determined by polymerase chain reaction (PCR), using the primers and methods described by Rigat and colleagues. Under some conditions, the ACE D allele amplifies more effectively than the longer I allele, resulting in mistyping of the ID as the DD genotype. Therefore, all DD genotypes were reconfirmed. Briefly, the sense primer used in the PCR was replaced with an insertion-specific primer that leads to selective amplification of the I allele and absence of the D allele in mistyped ID genotypes (Fig 1). No mistyping was identified.
Figure 1. Typical pictures of the PCR products in COPD patients with the II, ID, and DD genotypes.