ABSTRACT
Autophagosome formation and specific substrate recruitment during autophagy require ligation of the ubiquitin-like protein (UBL) Atg8 to the head group of the lipid phosphatidylethanolamine. Atg8 lipidation is mediated by distinctive UBL cascades involving autophagy-specific E1, E2, and E3 enzymes that differ substantially in sequence from components of other UBL conjugation cascades. Structural studies are important for elucidating the roles of Atg proteins that regulate multiple steps involved in autophagy. This chapter describes methods to prepare and crystallize selected proteins and complexes involved in autophagy UBL conjugation pathways, as a guide for strategies for structural and biochemical characterization of Atg proteins.
Subject(s)
Autophagy-Related Proteins/chemistry , Autophagy-Related Proteins/metabolism , Crystallography/methods , Autophagy , Autophagy-Related Protein 5/chemistry , Autophagy-Related Protein 5/genetics , Autophagy-Related Protein 5/isolation & purification , Autophagy-Related Protein 7/chemistry , Autophagy-Related Protein 7/metabolism , Autophagy-Related Proteins/isolation & purification , Crystallization/methods , Humans , Mutation , Protein Domains , Saccharomyces cerevisiae Proteins/chemistry , Saccharomyces cerevisiae Proteins/metabolism , Ubiquitin-Conjugating Enzymes/chemistry , Ubiquitin-Conjugating Enzymes/metabolism , Ubiquitin-Protein LigasesABSTRACT
PURPOSE: To assess in a consecutive series of patients, the diagnostic accuracy of dipyridamole echocardiography and to compare its prognostic power with that from other tests. METHODS: Seventy eight patients prospectively were underwent to dipyridamole echocardiography (DET) and coronary arteriography for diagnostic and post-infarction investigation. The majority was underwent to treadmill stress test and Tc99MIBI-dipyridamole planar myocardial scintigraphy. Eleven variables with possible prognostic importance were analyzed in a stepwise model to identify independent predictors of events. RESULTS: Sensitivity and specificity for the presence of coronary artery disease (in non-M patients) and multivessel disease (in post-MI patients) were, respectively, 44%, 91%, 53% and 88%. Mean follow-up was 21 +/- 9 months, during which there were four deaths, four unstable angina episodes, four nonfatal myocardial infarctions and eight revascularization procedures. By multivariate analysis, the only independent predictor of events was the change in wall motion score index from rest to peak stress. CONCLUSION: DET is a valuable alternative to exercise stress test and myocardial perfusion scintigraphy. Semiquantitative estimation of ischemic area during pharmacological stress is an important prognostic marker.
Subject(s)
Coronary Disease/diagnosis , Dipyridamole , Echocardiography/methods , Chi-Square Distribution , Coronary Angiography , Exercise Test , Female , Humans , Male , Middle Aged , Multivariate Analysis , Predictive Value of Tests , Prognosis , Prospective Studies , Radionuclide Angiography , Sensitivity and Specificity , Technetium Tc 99m Sestamibi , Vasodilator AgentsSubject(s)
Echocardiography , Endocarditis, Bacterial/diagnosis , Heart Valve Diseases/diagnosis , Adolescent , Adult , Diagnosis, Differential , Female , Humans , Male , Middle AgedABSTRACT
Foram analisados retrospectivamente vinte ecocardiogramas de pacientes com endocardite infecciosa ou historia pregressa desta doenca. A revisao da literatura permitiu extrair 609 ecocardiogramas de pacientes com endocardite, que serviram de fonte para uma serie de comparacoes e conclusoes. Verificou-se, desta forma, a incidencia de rotura de cordoalha tendinea valvar mitral, insuficiencia aortica, ecos anormais na via de saida do ventriculo esquerdo e seu significado e a presenca de anormalidades estruturais pre-existentes.As conplicacoes decorrentes da presenca de vegetacoes ao ecocardiograma foram estudadas, verificando-se nitido predominio de fenomenos embolicos, insuficiencia cardiaca e intervencao cirurgica naqueles pacientes com endocardite cujo ecocardiograma revelou a presenca de lesoes vegetantes. A contribuicao da ecocardiografia bidimensional foi, tambem, estudada, concluindo-se pela melhora da sensibilidade diagnostica com a associacao das duas tecnicas
