[Primary cutaneous malignant melanoma: retrospective studyfrom 1963 to 1997 at Hospital do Servidor Público Estadual de São Paulo]. / Melanoma maligno cutâneo primário: estudo retrospectivo de 1963 a 1997 no Hospital do Servidor Público Estadual de São Paulo.

OBJECTIVES: A retrospective study on the Primary Cutaneous Malignant Melanoma in the Hospital do Servidor Público Estadual de São Paulo (HSPE-SP) analyzing its distribution according to age, sex, race and cutaneous site. METHODOLOGY: We studied 222 patients with Primary Cutaneous Malignant Melanoma as diagnosed at Hospital do Servidor Público Estadual de São Paulo, Brazil between the period from 1963 to 1997. A retrospective study was performed. Data were expressed as inance of caucasians (98.19%) over afro-americans (1.81) and of the female sex (69.36%) over the male sex (30.63%) was found. The predominant age on the occasion of the diagnosis was between 50 and 60 years for the women (25.32%) and between 60 and 69 years for the men (22.52%). The most frequent site of the cancer in men was the back region (29.41%) and in the lower members in the women (38.31%). The most frequent level of the tumor invasion (Clark) was IV (39.77%), and the average of tumor thickeness (Breslow) was < 0.75 mm (28.4%). A 5 years survival was observed in 73.3% of the patients. CONCLUSIONS: At our Hospital the incidence of Primary Cutaneous Malignant Melanoma has shown an increase in recent years; these results are compatible with the most recent international surveys.

Melanoma maligno cutâneo primário: estudo retrospectivo de 1963 a 1997 no Hospital do Servidor Público Estadual de Säo Paulo / Primary cutaneous malignant melanoma occurence diagnosed at Hospital do Servidor Público Estadual de Säo Paulo between the period from 1963 through 1997

Objetivo. Estudar retrospectivamente a ocorrência do melanoma maligno cutâneo primário no nosso Hospital, objetivando analisar os dados referentes a sua distribuiçao pela idade, sexo, raça, localizaçao cutânea, sobrevida em cinco anos, nível de Clark, índice de Breslow e a taxa de incidência. Casuística e Método. Foram estudados 222 pacientes com melanoma maligno cutâneo primário, diagnosticados no Hospital do Servidor Público Estadual (FMO)-SP, entre os anos de 1963 a 1997. Elaborou-se um estudo do tipo retrospectivo. A descriçao dos dados foi expressa na forma de porcentagens. Para os cálculos dos dados epidemiológicos foram utilizados: Para taxa de incidência anual o número de casos novos no ano (numerador) e o número de consultas ambulatoriais em todo o Hospital no mesmo ano (denominador). Os dados restantes foram calculados por proporçao. A comparaçao entre as diferentes categorias se efetuou pelo teste de x2 e considerou-se significativo em p < 0,05. Resultados. Encontrou-se um predomínio de caucasóides (98,19 por cento) sobre negróides (1,81 por cento) e do sexo feminino (69,36 por cento) sobre o masculino (30,63 por cento). A idade predominante encontrada à época do diagnóstico foi entre 50 e 60 anos para as mulheres (25,32 por cento) e entre 60 a 69 anos para os homens (22,52 por cento). A localizaçao mais freqüente da neoplasia nos homens foi a regiao torácica posterior (29,41 por cento) e os membros inferiores nas mulheres (38,31 por cento). O nível de invasao tumoral (Clark) mais encontrado foi IV (39,77 por cento) e a espessura dos tumores (Breslow) mais freqüente foi <0,75mm (28,4 por cento). Tiveram sobrevida de cinco anos 73,3 por cento dos pacientes. Conclusoes. Evidenciou-se o aumento da ocorrência do melanoma maligno cutâneo primário ao longo destes anos no Hospital, sendo estes resultados compatíveis com os mais recentes trabalhos internacionais.

Molecular cloning of a cDNA for alpha-subunit of rat liver electron transfer flavoprotein.

Two cDNA clones for the alpha-subunit of rat liver electron transfer flavoprotein were isolated and their nucleotide sequences were determined. The longer cDNA contained a protein-coding region of 900 nucleotides and 3'-noncoding region of 335 nucleotides. The identity of the clone was confirmed by matching the amino acid sequence predicted from the cDNA with the sequence of one of the lysyl endopeptidase-digested peptides from the purified alpha-subunit. The molecular weight of the protein calculated from the protein-coding nucleotides was approx. 3,000 daltons smaller than that of the precursor, suggesting that the cDNA was not of full length. The derived amino acid composition fairly agreed with the chemically determined amino acid composition of the purified alpha-subunit, indicating that the protein-coding region contains most of the mature alpha-subunit.

Translation of messenger RNA of pig kidney D-amino acid oxidase in a cell-free system.

In vitro synthesis of D-amino acid oxidase [D-amino acid: O2 oxidoreductase (deaminating), EC 1.4.3.3], one of the peroxisomal flavin enzymes, was performed using a rabbit reticulocyte lysate system in order to elucidate the biosynthetic pathway of the enzyme. The apparent molecular weight of the synthesized enzyme protein was the same as that of D-amino acid oxidase purified from pig kidney. On the other hand, the enzyme protein was not detectable when a wheat germ lysate system was used for the translation. Denaturation of pig kidney poly(A)+ RNA with methylmercury hydroxide prior to the translation was found to enhance the synthesis of the enzyme protein. These results suggest a tight conformational structure of the mRNA used.

Evidence that a nicked C4b, C4b', is a functionally active C4b derivative.

Factor I-catalyzed C4b cleavage is a regulatory reaction for the classical pathway of the complement system. Although the reaction was shown to be a two-step reaction, production of a nicked form of C4b, C4b', as an intermediate cleavage product and subsequent splitting of C4b' into C4c and C4d, it is not known which of the two steps represents the inactivation of the C4b function in the assembly of C3 convertase, C4b,2a. We have purified C4b' and assessed the ability of C4b' to assemble C3 convertase with C2 by utilizing size exclusion high performance liquid chromatography. Evidence was obtained demonstrating that C4b' still retains the function of C4b to assemble C3 convertase. Thus, the substantial step for the inactivation of the C4b function appears to be the second cleavage reaction, that is, the cleavage of C4b' into C4c and C4d.

In vitro synthesis of pig kidney general acyl CoA dehydrogenase.

In vitro synthesis of general acyl CoA dehydrogenase [EC 1.3.99.3], one of the mitochondrial flavoenzymes, was carried out to elucidate its biosynthetic mechanism. Poly(A)+ RNA isolated from pig kidney was translated in vitro using wheat germ lysate system and the synthesized enzyme was immunoprecipitated by the antibody against purified pig kidney general acyl CoA dehydrogenase. The apparent molecular weight of the synthesized protein was estimated to be approximately 1,000 daltons larger than that of the mature enzyme, indicating that general acyl CoA dehydrogenase in pig kidney is synthesized as a precursor with a larger molecular weight.

Monoclonal anti-human C4b antibodies: stabilization and inhibition of the classical-pathway C3 convertase.

Two IgG mouse monoclonal antibodies (MAbs), Abs 242 and 463, were prepared by fusion of spleen cells from mice immunized with human C4b with a myeloma cell line, P3/ X 63-Ag 8.653. They were assessed for their effect on the activation and stability of the cell-bound classical-pathway C3 convertase, EAC14b2a and on the binding of C2 and C4bp to EC4b. Ab 242 recognized a conformational neoantigen which appeared upon activation of C4 with C-1s and disappeared after chain separation of C4b, while Ab 463 recognized a linear epitope in the beta-chain of C4b. Ab 242 was found to be a C4bp-like MAb: it accelerates the decay-dissociation of C3 convertase and interferes with the binding of C2 to C4b. It also interfered with the binding of C4bp to C4b. These results suggest that Ab 242 recognizes an epitope which is closely related to the C2- and C4bp-binding sites in C4b. Ab 463, on the other hand, was found to be a nephritic factor like MAb: it prolongs the half-life of C3 convertase from 8 to 30 min at 37 degrees C.

Human C4b-binding protein, C4bp. Chymotryptic cleavage and location of the 48 kDa active fragment within C4bp.

C4bp, a regulator of the classical pathway of complement system, is composed of 6-8 disulfide-linked subunit chains of 75 kDa. Upon incubation with chymotrypsin, C4bp was rapidly cleaved into a nicked C4bp, composed of disulfide-linked 48 kDa and 27 kDa fragments. Subsequent slow cleavage on the 27 kDa fragment resulted in the liberation of the active site-containing 48 kDa fragment from the nicked C4bp. The N-terminal amino acid sequence of the 48 kDa fragment was identical to that of the parent subunit chain of C4bp, indicating that the 48 kDa active fragment was released from the N-terminal side of the parent subunit chain. Based on these results, a possible gross structure of C4bp is proposed.

Limited chymotryptic cleavage of human C4-binding protein: isolation of a carbohydrate-containing core domain and an active fragment.

Human C4 binding protein (C4bp), which is a macromolecular weight (Mr 450,000-590,000) cofactor of C3b/C4b inactivator (I), is composed of 6 or 8 disulfide-linked polypeptide chains of Mr 75,000. Chymotrypsin cleaved C4bp into two major fragments; a large fragment of Mr 160,000, which contained carbohydrate chains and was composed of disulfide-linked polypeptide chains of Mr 25,000, and a small fragment of Mr 48,000, which was a single polypeptide chain and had the cofactor activity of C4bp. These results suggest that chymotrypsin liberates a functional domain-containing Mr 48,000 fragment from each subunit chain of C4bp and yields a core fragment derived from a disulfide-knot domain connecting each subunit chain of C4bp.

Cleavage of C4b by C3b inactivator: production of a nicked form of C4b, C4b', as an intermediate cleavage product of C4b by C3b inactivator.

We have investigated the mechanism of cleavage of C4b into C4c and C4d by the C3b inactivator (C3bINA) and revealed the formation of a nicked form of C4b as an intermediate cleavage product. The cleavage of C4b by the C3bINA was a two-step reaction. The first cleavage occurred on the alpha-chain (89,000 daltons) yielding two fragments, 73,000 daltons and 16,000 daltons. These fragments were bound to each other or to the beta or gamma chain through disulfide linkages. Therefore, an altered form of C4b, C4b', consisting of four disulfide-linked polypeptide chains with the same m.w. as C4b, was produced as an intermediate cleavage product. Subsequently, the second cleavage occurred on the alpha-chain fragment of 73,000 daltons to produce the two generally recognized fragments, C4c and C4d. In both cleavage reactions, a high m.w. cofactor protein, C4bC3bINACo, which is the same protein described as the C4 binding protein, was required, suggesting that both of the proteolytic processes are catalyzed by the C3bINA.