Graduaçäo médica e especializaçäo: uma incompatibilidade aparente / Medical graduation and specialization: an apparent incompatibility

Objetivo. Este artigo apresenta resultados parciais da pesquisa, desencadeada a partir de 1989, de avaliaçäo continuada do ensino de graduaçäo médica da Escola Paulista de Medicina (EPM), com a qual se implantou amplo processo de avaliaçäo institucional. Metodologia. O estudo, de base amostral, envolve o levantamento de expectativas e opiniöes de docentes, alunos e egressos, constituindo três subprojetos específicos. Resultados. Os autores chamam a atençäo para näo-terminalidade da formaçäo médica na EPM, levando em conta que os egressos näo entram no mercado de trabalho ao final da graduaçäo. Conclusäo. Este resultado aponta para a necessidade de reflexäo em torno do significado da näo-terminalidade por referência ao longo processo de formaçäo médica. Neste caso, a característica apontada näo está associada à ausência de qualidade e, sim, à incorporaçäo, no currículo de graduaçäo, do desenvolvimento de técnicas e procedimentos profissionais que conduzem à inexorável especializaçäo do conhecimento, atingida somente por meio de formaçäo pós-graduada.

Purification and characterization of endopeptidase H2, a kinin inactivating serine proteinase (kininase) from human urine

1. A kinin-inactivating chymotrypsin-like serine-endopeptidase was purified 202-fold from human urine by DEAE-cellulose chromatography, gel filtration, DEAE/HPLC chromatography and affinity chromatography. It hydrolyzed bradykinin at the Phe5-Ser6 peptide bond at a rate of 1.090 mumol min-1 mg protein-1 at pH 8.0 and 37 degrees C. The molecular weight of this endopeptidase H2, estimated by SDS-polyacrylamide gel electrophoresis and by gel filtration, was 60 kDa, and its optimum pH for bradykinin hydrolysis was near 8.5. 2. Bradykinin inactivating activity was inhibited 100 per cent by the serine-proteinase inhibitor PMFS (1 mM) and the chymotrypsin inhibitor TPCK (5 mM). Reagents such as 2-mercaptoethanol (3 mM) and pOH-mercuribenzoate (3 mM) inhibited the enzyme by 100 per cent and 67 per cent , respectively. 3. Endopeptidase H2 hydrolyzes the Phe-Ser bond of peptides related to bradykinin and its activity appears to be limited to peptide chains of < or = 18 amino acid residues since it does not hydrolyze BAM 22, peptide E or kininogen. 4. The molecular size and inhibition profile suggested that endopeptidase H2 differs from the serine-proteinases previously described in rat liver, rat hepatic endothelium, rat and rabbit brain. 5. The physiological role of endopeptidase H2 may be a link between the kinin and neuropeptide systems in the control of water-electrolyte balance

Purification and partial characterization of Enterolobium contortisiliquum seed arylamidase

Arylamidase activity was isolated from Enterolobium contortisiliquum seeds (2 U/g) using L-Leu-2-naphthlamide as substrate to monitor the prification. The enzyme preparation was purified 733-fold by ammonium sulfate precipitation, and by ion eschange and gel filtration chromatography, in 6,6% yield. SDS-Polyacrylamide gel electrophoresis after fast protein liquid chromatography on a Mono Q column, showed only one protein band with a molecular weight of 35 kDa. The optimum pH for arylamidase activity was 6.5. Taking the hydrolysis rate of Lys-2-naphthylamide as one, the relative rates at which the other substrates were hydrolyzed were: Leu-2-naphthlamide, 30, Met-2-naphthlamide, 18, Arg-2-naphthlamide, 2, Ala-2-naphthylamide, 1.5, and L-Leu-p-nitroanilide, 26. The arylamidase activity was inhibited 50% by 0.1 mM HgCl2, 0.1 mM ZnCl2, 0.13 mM NiCl2, 0.2 mM o-phenanthroline and 1 * M soidum p-hydroxymercuribenzoate, and activated 35% by 5.0 * M EDTA. Iodoacetate (0.067 mM), dithioerythritol and 2-mercaptoethanol (3.3 mM), and chloride ions (0.2 M) had no effect on the enzyme activity