ABSTRACT
RESUMO Cyrtocymura scorpioides (sin. Vernonia scorpioides (Lam.) Pers.), Piracá é utilizada popularmente para tratamento de úlceras, traumatismos, candidíase, processos inflamatórios e dores musculares. Objetivou-se verificar nas plantas cultivadas na Vila Nair, Jardim São Dimas e Urbanova em São José dos Campos - SP, a influência da poluição veicular nos rendimentos da matéria seca (folhas), no óleo essencial, e no extrato bruto, bem como a ação citotóxica em células HEP-2 e L929, e identificar os componentes do óleo essencial e ação fungicida em Candida albicans. As estacas (54) foram cultivadas durante 6 meses em solo + adubo (2:1) na Universidade do Vale do Paraíba - UNIVAP, e distribuídas nas estações Dutra (E1 - tráfego intenso), Teotônio (E2 - tráfego médio) e Urbanova (E3 - tráfego baixo), onde 18 mudas foram cultivadas durante 6 meses, sendo 3 repetições de 6 plantas. O óleo essencial foi extraído por hidrodestilação e seus componentes identificados por cromatografia gasosa acoplado a espectrômetro de massas (CG-MS), através de indice de similaridade com a base de espectros Wiley L. O extrato bruto foi concentrado por rotavapor. A ação fúngica foi avaliada pelo teste de difusão em disco e a citotoxicidade pelo teste MTT. Em Urbanova (E3) verificouse maior rendimento da matéria seca, do extrato bruto e do óleo essencial. Identificou-se no óleo essencial: ß-cariofileno, α-cariofileno, germacreno D, delta-cadineno e cariofileno. O Óleo Essencial possui possui baixa ação fungicida em C. albicans, enquanto o extrato hidroalcóolico se mostrou citotóxico para L929 e HEp-2.
ABSTRACT Cyrtocymura scorpioides (syn. Vernonia scorpioides (Lam.) Pers.), known as Piracá, is popularly used for the treatment of ulcers, trauma, candidiasis, inflammatory disorders, and muscle pain. This study aimed to assess the influence of vehicular pollution on the yield of dry matter (leaves), essential oil, and crude extract, and the cytotoxic action in HEP-2 and L929 cells. This study also aimed to identify the components of the essential oil, and verify its fungicidal action against Candida albicans in plants grown in Vila Nair, Jardim São Dimas, and Urbanova, São José dos Campos - SP, Brazil. The seedlings (54) were grown in soil + fertilizer (2:1) at the Universidade do Vale do Paraiba - UNIVAP, and distributed to different stations, Dutra (E1 - heavy traffic), Teotônio (E2 - medium traffic), and Urbanova (E3 - low traffic), where 18 seedlings were cultivated for 6 months, with 3 replicates of 6 plants. The essential oil was extracted by hydrodistillation and its components were identified by by Gas chromatography - mass spectrometry (GC-MS), with a similarity index computed using the Wiley L spectra. The crude extract was concentrated in a Buchi Rotary Evaporator R-114, the fungicidal action and cytoptoxicity were evaluated using the disk diffusion method and the MTT test, respectively. In Urbanova (E3), high yields of dry matter, crude extract, and essential oil were obtained. The following components were identified in the oil: ß-caryophyllene, α -caryophyllene, germacrene D, delta-cardinene, and caryophyllene oxide. The oil was found to have low fungicidal action against C. albicans, while the hydroalcoholic extract was cytotoxic to L929 and HEP-2.
Subject(s)
Vernonia/classification , Environmental Pollution/adverse effects , Oils, Volatile/classification , Cytotoxins/analysisABSTRACT
The gp43 glycoprotein is an immune-dominant antigen in patients with paracoccidioidomycosis (PCM). It is protective against murine PCM and is a putative virulence factor. The gp43 gene of Paracoccidioides brasiliensis B-339 is located in a 1,329-bp DNA fragment that includes two exons, a 78-bp intron, and a leader peptide-coding region of 105 bp. Polymorphism in gp43 has been suggested by the occurrence, in the same isolate or among different fungal samples, of isoforms with distinct isoelectric points. In the present study we aligned and compared with a consensus sequence the gp43 precursor genes of 17 P. brasiliensis isolates after sequencing two PCR products from each fungal sample. The genotypic types detected showed 1 to 4 or 14 to 15 informative substitution sites, preferentially localized between 578 and 1166 bp. Some nucleotide differences within individual isolates (noninformative sites) resulted in a second isoelectric point for the deduced protein. The most polymorphic sequences were also phylogenetically distant from the others and encoded basic gp43 isoforms. The three isolates in this group were from patients with chronic PCM, and their DNA restriction patterns were distinct in Southern blots. The nucleotides encoding the inner core of the murine T-cell-protective epitope of gp43 were conserved, offering hope for the development of a universal vaccine.
Subject(s)
Fungal Proteins , Glycoproteins/genetics , Immunodominant Epitopes/genetics , Oligosaccharides/genetics , Paracoccidioides/immunology , Paracoccidioidomycosis/microbiology , Polymorphism, Genetic/genetics , Animals , Antigens, Fungal/genetics , Blotting, Southern , Humans , Molecular Sequence Data , Paracoccidioides/genetics , Paracoccidioides/isolation & purification , Phylogeny , Polymerase Chain Reaction/methods , Sequence Analysis, DNAABSTRACT
BACKGROUND: The liver inactivates considerable amounts of bradykinin; the main liver kinin-inactivating enzyme (BIE, bradykinin inactivating endopeptidase) hydrolyses specifically the Phe5-Ser6 bond of the nonapeptide and it has been characterized as the oligoendopeptidase E.C.3.4.24.15. When orthotopic liver transplantation is performed there is a correlation between the increase of amino acid concentration in the preservation fluid (as a consequence of proteolysis) and graft dysfunction. AIM: Verify if BIE is released from livers stored ex vivo. METHOD: Wistar rats (180-220 g) livers were exsanguinated and after removal were preserved in Braun Collins fluid or Krebs solution at 4 degrees C. Aliquots were collected from the preservation fluid at 0, 4, 8, 24 h, for ALT, AST, LDH and BIE assays. The fluorimetric activity of BIE was assayed upon Abz-RPPGFSPFRQ-EDDnp (synthetic BK analogue) and its presence was confirmed by immunoblotting, revealed with specific antibody anti-E.C.3.4.24.15. RESULTS: The release of ALT, AST, LDH and BIE is significant between 8-24 h. In the 24 h aliquots the four enzymes concentration increased in the Braun Collins fluid 8, 7, 19 and 10 respectively, and in the Krebs solution 21, 17, 27, 21 respectively, when compared to the zero time aliquot activities. The ratio ALT/LDH was always < 1. CONCLUSION: There is BIE release during ex vivo liver storage; this information may be useful as an indicator of the graft preservation condition; a decrease of the liver kinin-inactivating capability could affect the graft vascular reactivity.
Subject(s)
Bradykinin/antagonists & inhibitors , Endopeptidases/metabolism , Endopeptidases/pharmacology , Liver/enzymology , Animals , Organ Preservation , Rats , Rats, WistarABSTRACT
Objetivo - Ofígado inativa quantidades consideráveis de bradicinina; a principal enzima hepática cinino-inativadora (BIE, bradykinin inativating endopeptidase) hidrolisa especificamente a ligaçao Phe-Ser do nonapeptídio e foi caracterizada como sendo a oligoendopeptidase EC 3.4. 24.15. No transplante ortotópico de fígado existe correlaçao entre aumento da concentraçao de aminoácidos no líquido de preservaçao (conseqUência de proteólise) e falência do enxerto. O objetivo deste trabalho é verificar se ocorre liberaçao da BIE de fígados preservados ex-vivo no líquido Braun-Collins ou em soluçao de Krebs-Henseleit bicarbonato (Krebs). Método. Fígado de ratos Wistar (180-220g) foram exsangüinados e a pós remoçao foram preservados em líquido Braun Collins ou em soluçao Krebs, a 4 graus Celsius. Foram retiradas alíquotas do líquido de preservaçao nos tempos 0, 4, 8 e 24 horas, para dosagem de ALT, AST, DHL e BIE. A atividade fluorimétrica da BIE foi ensaiada com o substrato Abz-RPPGFSPFRQ-EDDnp (análogo sintético da bradicinina) e sua presença confirmada por immunoblotting, revelado com anticorpo específico anti-EC 3.4.24.15. Resultados. A liberaçao de ALT, AST, DHL e BIE é significativa no período 8-24 hs. Nas alíquotas de 24 hs, em relaçao ao tempo zero, a concentraçao das quatro enzimas aumentou, respectivamente, no líquido Braun Collins, 8, 7, 19 e 10 vezes e, na soluçao de Krebs, 21, 17, 27 e 21 vezes; a relaçao ALT/DHL foi sempre inferior a um. Conclusao. Ocorre liberaçao de BIE durante a preservaçao ex-vivo do fígado, o que poderá servir como indicaçao da condiçao de preservaçao do enxerto; diminuiçao da capacidade cinino-inativadora do fígado poderá afetar sua reatividade vascular.
