RESUMO
PURPOSE: The aim of the study was to investigate the management and prognosis of Pure primary squamous cell carcinoma (PPSCC) of the breast. MATERIALS AND METHODS: This study is a multicentre retrospective cohort from three French tertiary referral hospitals (Rennes, Orléans and Tours) including all women treated for a PPSCC of the breast defined by squamous cells that could contain a minority of sarcomatoid component. We excluded carcinomas with a ductual component. Clinicopathologic, radiological and therapeutic patterns were described. Demographic, histological and therapeutic characteristics were compared to a population of women with triple negative invasive breast carcinomas. RESULTS: Twelve patients were included, with a mean age of 71.6 years. All lesions were unifocal, with a cystic complex ultrasound mass in 50% of cases. Mean tumor size was 43mm, with axillary lymph node metastasis in 25% of patients. The comparison with a population of women with triple negative breast carcinomas revealed that women with PPSCC were older (71 versus 57 years, p=0.003), tumor size was larger (43mm versus 25mm, p=0.032) and local recurrence occurred earlier (three months versus 38 months, p=0.014). CONCLUSION: PPSCC is a rare entity with a worse prognosis in comparison with triple negative invasive carcinoma.
Assuntos
Neoplasias da Mama/terapia , Carcinoma de Células Escamosas/terapia , Recidiva Local de Neoplasia , Neoplasias de Mama Triplo Negativas/terapia , Adulto , Idoso , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/epidemiologia , Neoplasias da Mama/patologia , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/epidemiologia , Carcinoma de Células Escamosas/patologia , Feminino , França/epidemiologia , Humanos , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos , Neoplasias de Mama Triplo Negativas/diagnóstico , Neoplasias de Mama Triplo Negativas/epidemiologia , Neoplasias de Mama Triplo Negativas/patologiaRESUMO
383 psychotics patients were selected in 1975 according to the french classification (INSERM). These initial population was simultaneously subdivided according 2 types of criteria: Diagnosis criteria (DSM III Axis 1), Psychosocial adaptation criteria (DSM III axis V). For each subjects we investigates in 1985 evolution of following data previously collected in 1975: Diagnosis and psychosocial adaptation, Marital status and presence of social welfare work, Treatment variable (Hospitalization and out patient department care, neuroleptic treatment). Results indicate than: psychosocial adaptation criteria are better than diagnosis criteria for outcome judgment. most of the time there is no correlation between high quantity of care and better outcome, furthermore some time there is a correlation between high quantity of care and negative outcome, We must consider, as one explanation of this fact, that it's the more severe patients who use the higher quantity of care.
Assuntos
Avaliação de Processos e Resultados em Cuidados de Saúde , Transtornos Psicóticos/classificação , Adaptação Psicológica , França , Humanos , Casamento , Prognóstico , Transtornos Psicóticos/diagnóstico , Transtornos Psicóticos/psicologia , Serviço Social em PsiquiatriaRESUMO
Nanogram amounts of soluble aminopeptidases, dehydrogenases, and esterases were detected by nondenaturing ultralow gelling point agarose-polyacrylamide gel electrophoresis (ULGA-PAGE). Cytosolic fractions from Frankia sp. were electrophoresed at 4 degrees C in the presence of Co2+, Zn2+, or Mg2+ ions. Then, aminopeptidases and esterases were revealed by simultaneous capture staining by using fast garnet GBC diazonium salt as the chromogenic coupling compound. Dehydrogenases were revealed by using nitro blue tetrazolium salt as electron acceptor. A variety of aminopeptidases, dehydrogenases, and esterases could be identified by their migration in ULGA-PAGE and by their sensitivities to NaCl, CoSO4, ZnSO4, and MgCl2 when assayed "ingel." The presence of agarose was essential for the detection of the complex enzyme patterns. The patterns were remarkably similar for the five Frankia strains isolated from Allocasuarina and Casuarina host plants and differed from those of Frankia strains isolated from Comptonia and Hippophaë host plants. A nomenclature is proposed for aminopeptidases and other Frankia enzymes. The richness of the Frankia amino-peptidases and esterases zymograms makes them adequate marker enzymes for taxonomical, genetic, or biochemical studies. Dehydrogenases might also be useful, although a more restricted number of bands were found with L-lactic and L-malic acid as substrates.
Assuntos
Actinomyces/enzimologia , Aminopeptidases/análise , Esterases/análise , Oxirredutases/análise , Actinomyces/crescimento & desenvolvimento , Citosol/enzimologia , Eletroforese em Gel de Ágar , Eletroforese em Gel de Poliacrilamida , Sensibilidade e Especificidade , Especificidade por Substrato , Terminologia como AssuntoRESUMO
To investigate protein secretion by the nitrogen-fixing actinomycete Frankia isolate BR, we designed a rapid DEAE adsorption, salt elution and Biogel P6DG desalination method to concentrate protein from the growth medium. Secreted proteins reached a maximum concentration (5.6 gm l-1) in the medium at growth arrest. Analysis by SDS-PAGE detected up to 63 extracellular polypeptides when Frankia cells were grown under stirred conditions in BAP medium supplemented with phosphatidylcholine and MES buffer and 65 proteins in stirred BAP media alone. The pattern of extracellular polypeptides changed during growth. Several extracellular proteolytic activities were detected and compared with intracellular ones. The substrate specificity of the extracellular and intracellular aminopeptidase activities were the same. Also, the electrophoretic migration patterns of secreted and intracellular aminopeptidases could not be distinguished. Secretion of the proline-specific aminopeptidase FAP proteinase (PF) were secreted: 10 had the same electrophoretic mobility as their intracellular counterparts after SDS-gelatine-PAGE while five (PF - 39.5, PF - 38.5, PF - 36.5, PF - 25.5 and PF - 20.5 kDa) had a different electrophoretic mobility and, therefore, appeared to be exclusively extracellular. At least seven extracellular proteinases appeared to increase coordinately in activity shortly before growth arrest.
Assuntos
Aminopeptidases/metabolismo , Proteínas de Bactérias/metabolismo , Endopeptidases/metabolismo , Frankia/fisiologia , Divisão Celular/fisiologia , Meios de Cultura , Eletroforese , Frankia/crescimento & desenvolvimento , Fatores de TempoRESUMO
We have recently described the presence of a high molecular mass multicatalytic proteinase complex (megaproteinase; 28 S, 1300 kDa) in Frankia strain BR. The complex dissociates into 11 low molecular mass proteinase subunits (40-19 kDa) when subjected to sodium dodecyl sulfate - gelatin - polyacrylamide gel electrophoresis. We show here that the activity of these proteinase subunits strongly increased after cessation of growth in stirred BAP-PCM mineral medium. Subsequent addition of either BAP medium components or sodium propionate alone, as carbon source, to a Frankia culture at the end of the exponential growth phase was found to prolong growth for 1 additional day, and to delay the increase in activity of the proteinase subunits for 3 days after cessation of growth. Addition of ammonium chloride alone, as nitrogen source, had no effect. On the other hand, when Frankia cells in the late exponential phase (3 days) were resuspended in a culture filtrate recovered from a 5-day-old culture and supplemented with BAP-PCM medium components, the biomass yield decreased to about 50%. Also, the activity of the proteinase subunits increased as soon as growth ceased. The ability of this culture filtrate to inhibit growth and stimulate the activity of proteinase subunits was partially lost by heating or was largely removed by DEAE-cellulose treatment. Thus, our findings indicate an extracellular control of Frankia megaproteinase activity, suggesting that carbon source depletion and probably accumulation of heat-sensitive growth-inhibiting metabolites in the medium are determining factors.
Assuntos
Carbono/metabolismo , Endopeptidases/metabolismo , Frankia/enzimologia , Divisão Celular , Meios de Cultura , Ativação Enzimática , Frankia/crescimento & desenvolvimento , Complexos Multienzimáticos/metabolismo , Estrutura Terciária de ProteínaRESUMO
A major-high-molecular mass proteinase and seven latent minor proteinases were found in cell extracts and in concentrates of culture medium from Frankia sp. strain BR after nondenaturing electrophoresis in mixed gelatin-polyacrylamide gels. All of these complexes showed multicatalytic properties. Their molecular masses and their sedimentation coefficients varied from 1,300 kDa (28S) to 270 kDa (12S). The electroeluted 1,300-kDa proteinase complex dissociated into 11 low-molecular-mass proteinases (40 to 19 kDa) after sodium dodecyl sulfate activation at 30 degrees C and electrophoresis under denaturing conditions. All of these electroeluted proteinases hydrolyzed N-carbobenzoxy-Pro-Ala-Gly-Pro-4-methoxy-beta- naphthylamide, D-Val-Leu-Arg-4-methoxy-beta-naphthylamide, and Boc-Val-Pro-Arg-4-methyl-7-coumarylamide, whereas Suc-Leu-Leu-Val-Tyr-4-methyl-7-coumarylamide was cleaved only by the six lower-molecular-mass proteinases (27.5 to 19 kDa). Examination by electron microscopy of uranyl acetate-stained, electroeluted 1,300- and 650-kDa intracellular and extracellular proteinase complexes showed ring-shaped and cylindrical particles (10 to 11 nm in diameter, 15 to 16 nm long) similar to those of eukaryotic prosomes and proteasomes. Polyclonal antibodies raised against rat skeletal muscle proteasomes cross-reacted with all of the high-molecular-mass proteinase complexes and, after denaturation of the electroeluted 1,300-kDa band, with polypeptides of 35 to 38, 65, and 90 kDa. Electrophoresis of the activated cell extracts under denaturing conditions revealed 11 to 17 gelatinases from 40 to 19 kDa, including the 11 proteinases of the 1,300-kDa proteinase complex. The inhibition pattern of these proteinases is complex. Thiol-reactive compounds and 1-10-phenanthroline strongly inhibited all of the proteinases, but inhibitors against serine-type proteinases were also effective for most of them.
Assuntos
Actinomycetales/enzimologia , Cisteína Endopeptidases/metabolismo , Endopeptidases/metabolismo , Complexos Multienzimáticos/metabolismo , Centrifugação com Gradiente de Concentração , Reações Cruzadas , Cisteína Endopeptidases/efeitos dos fármacos , Cisteína Endopeptidases/imunologia , Cisteína Endopeptidases/ultraestrutura , Eletroforese em Gel de Poliacrilamida , Endopeptidases/química , Endopeptidases/efeitos dos fármacos , Endopeptidases/imunologia , Endopeptidases/ultraestrutura , Ativação Enzimática , Gelatina/metabolismo , Substâncias Macromoleculares , Microscopia Eletrônica , Complexos Multienzimáticos/efeitos dos fármacos , Complexos Multienzimáticos/imunologia , Complexos Multienzimáticos/ultraestrutura , Coloração Negativa , Inibidores de Proteases/farmacologia , Complexo de Endopeptidases do Proteassoma , Frações Subcelulares/enzimologia , Especificidade por SubstratoRESUMO
The complete genomic organization of the Drosophila troponin T (TnT) gene shows many interesting features, including the presence of a microexon of only 3 nucleotides conserved among Drosophilidae. It is the smallest bona fide exon so far described, placing a new lower limit on the nucleotide number required for correct splicing. Four muscle-type specific transcripts are generated by developmentally regulated alternative splicing. Exons 3, 4, and 5 are absent in the transcript present in jump and flight muscles. A total of 11 exons are present in the adult hypodermic muscles transcript, whereas the microexon is absent in the larval hypodermic musculature. The two isoforms differ in a lysine residue. Post-translational regulation of the flight muscles/tergal depressor of the trochanter-specific isoform is involved in flight and/or jump function. The interaction domains of TnT in the tropomyosin-troponin complex are strongly conserved in the known vertebrate and invertebrate TnT sequences, whereas the terminal regions show an important variability. The COOH-terminal region shows important phylogenetic variations, whereas the NH2-terminal domain is associated with specific muscle types in a particular organism, a finding that discloses a selective value for these domains in the functionality of distinct muscles in different organisms.
Assuntos
Troponina/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Mapeamento Cromossômico , Drosophila melanogaster , Éxons , Hibridização In Situ , Dados de Sequência Molecular , Regiões Promotoras Genéticas , RNA Mensageiro/metabolismo , Troponina TRESUMO
Computational analysis predicted three potential hydrophobic transmembrane alpha-helices within the Ura2 multidomain protein of Saccharomyces cerevisiae, the C-terminal subdomain of which catalyses the second step of uridine-monophosphate biosynthesis by its L-aspartate carbamoyltransferase activity (EC 2.1.3.2). The fourth step of pyrimidine biosynthesis is catalysed by dihydro-orotate dehydrogenase (Ura1 protein; EC 1.3.99.11), which was similarly characterized as a peripheral membrane protein. Ex situ, the activities of the investigated enzymes were associated both with isolated yeast membranes, fractionated by differential centrifugation to remove intact nuclei, and with soluble cytoplasmic proteins.
Assuntos
Aspartato Carbamoiltransferase/metabolismo , Carbamoil Fosfato Sintase (Glutamina-Hidrolizante)/metabolismo , Complexos Multienzimáticos/metabolismo , Oxirredutases atuantes sobre Doadores de Grupo CH-CH , Oxirredutases/metabolismo , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/enzimologia , Sequência de Aminoácidos , Aspartato Carbamoiltransferase/biossíntese , Carbamoil Fosfato Sintase (Glutamina-Hidrolizante)/biossíntese , Membrana Celular/enzimologia , Citoplasma/enzimologia , Di-Hidro-Orotato Desidrogenase , Corantes Fluorescentes/química , Indóis/química , Dados de Sequência Molecular , Complexos Multienzimáticos/biossíntese , Oxirredutases/biossíntese , Estrutura Secundária de Proteína , Saccharomyces cerevisiae/fisiologiaRESUMO
The Ura2 multidomain protein catalyses the first two steps of pyrimidines biosynthesis in Saccharomyces cerevisiae. It consists of a 240 kDa polypeptide which contains carbamyl phosphate synthetase and aspartate transcarbamylase domains. The Ura2 protein was believed to be nucleoplasmic, since one of the aspartate transcarbamylase reaction products, monophosphate, was reported to be precipitated by lead ions inside nuclei. However, this ultracytochemical approach was recently shown to give artifactual lead polyphosphate precipitates, and the use of cerium instead of lead failed to reveal this nucleoplasmic localization. Ura2 localization has therefore been undertaken by means of three alternative approaches based on the detection of the protein itself: (a) indirect immunofluorescence of yeast protoplasts; (b) immunogold labelling of ultrathin sections of embedded yeast cells (both approaches using affinity purified primary antibodies directed against the 240 kDa Ura2 polypeptide chain, or against a 22 residue peptide specific of the carbamyl phosphate synthetase domain); and (c) direct fluorescence of cells expressing an Ura2-green fluorescent protein hybrid. All three approaches localize the bulk of Ura2 to the cytoplasm, whereas the signals associated with the nucleus, mitochondria or vacuoles are close to or at the background level.