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1.
Acta Virol ; 62(3): 310-325, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30160147

RESUMO

Respiratory syncytial virus (RSV) is the major cause of acute lower respiratory tract infection in infants. Winter outbreaks in Chile result in 5% of infected children hospitalized, with 0.01% mortality. Increased evidence indicates that viral and host factors modulate the severity of infection. Using DNA microarrays, we characterized the genome-wide transcriptional response of lung mucoepidermoid cells (NCI-H292) at 0, 24, 48, 72 and 96 hours post-infection (hpi) with a single dose of RSV/A. During the whole studied period, a bi-phasic gene expression profile was observed by a total of 330 differentially expressed genes. About 60% of them were up-regulated between 24-72 hpi and then turned-off at 96 hpi. This transient, early gene expression pattern was significantly enriched in biological processes like interferon signaling, antigen processing and presentation, double-stranded RNA binding and chemokine activity. We detected 27 common genes up-regulated between 24-72 hpi, from which IFIT1, IFI44, MX1, CXCL11 and OAS1 had the highest expression. The second pattern comprised over 120 genes, which remained silenced until 72 hpi, but were steeply up-regulated by 96 hpi. Biological processes of this late-response profile included cell cycle division and microtubule cytoskeleton organization. Conversely, the genes belonging to virus response pathway showed a decreased expression at 96 hpi. We conclude that RSV induces an early innate immune activation profile response until 72 hpi. Thereafter, the viral response is inhibited, leading to host cell recovery. The presented cellular model allows to study the specific pathways involved in elimination of infection at prolonged time intervals and their subsequent analysis in severe RSV disease of infants and/or older adults.


Assuntos
Células Epiteliais/metabolismo , Infecções por Vírus Respiratório Sincicial/genética , Vírus Sincicial Respiratório Humano/fisiologia , Células Epiteliais/virologia , Perfilação da Expressão Gênica , Humanos , Pulmão/metabolismo , Pulmão/virologia , Infecções por Vírus Respiratório Sincicial/metabolismo , Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sincicial Respiratório Humano/genética , Vírus Sincicial Respiratório Humano/isolamento & purificação , Fatores de Tempo , Transcriptoma
2.
Oral Dis ; 21(6): 730-8, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25757505

RESUMO

OBJECTIVES: To evaluate the expression and localization of MUC1/SEC and MUC1/Y isoforms in labial salivary glands (LSG) from Sjögren's syndrome patients (SS patients), as well as their in vitro expression induced by cytokines. SUBJECTS AND METHODS: Labial salivary gland from 27 primary SS patients and 22 non-SS sicca subjects were studied. Relative MUC1/SEC and MUC1/Y mRNA levels were determined by qPCR and protein levels by Western blotting. Induction of mucin mRNAs was assayed in vitro. Immunohistochemistry was used for localization. RESULTS: Relative MUC1/SEC and MUC1/Y mRNA and protein levels were significantly higher in LSG from SS patients. These mRNAs were induced by cytokines. MUC1/SEC and MUC1/Y were detected in acini apical region of control LSGs, and significant cytoplasmic accumulation was observed in acini of SS patients. MUC1/Y localized in acinar nuclei and cytoplasm of inflammatory cells of LSG from SS patients. A strong positive correlation was observed between cellular MUC1/SEC levels and glandular function determined by scintigraphy. CONCLUSIONS: We show for the first time that MUC1/SEC and MUC1/Y are expressed in LSG of both SS patients and non-SS sicca subjects. The observed overexpression and aberrant localization of MUC1/SEC and MUC1/Y and their induction by pro-inflammatory cytokines may favor the perpetuation of the inflammatory environment that disrupts the salivary glandular homeostasis in SS patients.


Assuntos
Mucina-1/genética , Mucina-1/metabolismo , RNA Mensageiro/metabolismo , Síndrome de Sjogren/genética , Síndrome de Sjogren/metabolismo , Células Acinares/química , Adulto , Idoso , Estudos de Casos e Controles , Núcleo Celular/química , Células Cultivadas , Citocinas/farmacologia , Citoplasma/química , Feminino , Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Pessoa de Meia-Idade , Mucina-1/análise , Isoformas de Proteínas/análise , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Glândulas Salivares Menores/química , Glândulas Salivares Menores/metabolismo , Adulto Jovem
3.
J Autoimmun ; 42: 7-18, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23497939

RESUMO

The most difficult component in our understanding of human autoimmunity remains a rigorous dissection of etiological events. Indeed, the vast literature on autoimmune diseases focuses on the inflammatory response, with the hope of developing drugs that reduce inflammation. However, there is increasing recognition that understanding the immunobiology of target tissues will also have direct relevance to disease natural history, including breach of tolerance. Sjögren's syndrome is essentially an epitheliitis and there are major changes to normal architectural salivary organization. We propose that loss of homeostasis is the initial event that precipitates inflammation and that such inflammatory response includes not only the adaptive response, but also an intense innate immune/bystander response. To understand these events this review focuses on the architecture, phenotype, function and epithelial cell organization. We further submit that there are several critical issues that must be defined to fully understand epithelial cell immunobiology in Sjögren's syndrome, including defining epithelial cell polarity, cell-cell and cell to extracellular matrix interactions and a variety of chemical and mechanical signals. We also argue that disruption of tight junctions induces disorganization of the apical pole of salivary acinar cells in Sjögren's syndrome. In addition, there will be a critical role of inflammatory cytokines in the apico-basal relocation of tight junction proteins. Further, the altered disorganization and relocation of proteins that participate in secretory granule formation are also dysregulated in Sjögren's syndrome and will contribute to abnormalities of mucins within the extracellular matrix. Our ability to understand Sjögren's syndrome and develop viable therapeutic options will depend on defining these events of epithelial cell biology.


Assuntos
Células Acinares/imunologia , Células Epiteliais/imunologia , Proteínas SNARE/imunologia , Síndrome de Sjogren/imunologia , Junções Íntimas/imunologia , Animais , Adesão Celular , Polaridade Celular , Citocinas/imunologia , Exocitose , Matriz Extracelular/metabolismo , Homeostase , Humanos , Mediadores da Inflamação/imunologia , Mucinas/metabolismo
4.
Horm Metab Res ; 44(9): 656-61, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22411587

RESUMO

Due to its ability to induce vascular endothelial growth factor expression and proliferation, migration, and vasculogenesis of endothelial cells, nerve growth factor (NGF) has been considered as an angiogenic factor in epithelial ovarian cancer (EOC). In this work, we evaluated the angiogenic and proliferative mRNA expression profiles of EOC and addressed the responsiveness of EOC explants to NGF stimulation. Twenty EOC samples were obtained from Obstetrics and Gynecology Department, University of Chile's Clinical Hospital. Global gene expression profiles of selected poorly differentiated serous EOC samples were obtained with DNA oligonucleotide microarrays. In addition, EOC explants were subjected to NGF stimulation and levels of p-AKT, BAX, BCL2, Ki-67, c-MYC, and FOXL2 proteins were determined by immunohistochemistry. Results showed that mRNAs coding for specific transcriptional regulators and antiapoptotic components of the NGF signaling pathway were upregulated in EOC cells. At the protein level, key members of the NGF pathway including p-AKT, BCL2/BAX, Ki-67, and c-MYC were found increased, while FOXL2 was decreased in response to NGF stimulation. These findings strongly suggest that NGF stimulates cellular proliferation of human EOC.


Assuntos
Proliferação de Células , Neoplasias Epiteliais e Glandulares/metabolismo , Neoplasias Epiteliais e Glandulares/fisiopatologia , Fator de Crescimento Neural/metabolismo , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/fisiopatologia , Idoso , Carcinoma Epitelial do Ovário , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Pessoa de Meia-Idade , Neoplasias Epiteliais e Glandulares/genética , Fator de Crescimento Neural/genética , Neoplasias Ovarianas/genética , Transdução de Sinais , Regulação para Cima
5.
World J Microbiol Biotechnol ; 28(3): 1107-13, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22805832

RESUMO

The yeast Saccharomyces cerevisiae is the main microorganism responsible for wine fermentation and its development influences the quality of wine. A problem affecting these types of fermentations, generating important losses in this industry, are the slow or stuck fermentations which may result from low nitrogen availability in the must. Therefore, several studies have been directed towards identifying genes involved in nitrogen metabolism using high throughput strategies which include subjecting the yeast to changes in the type or concentration of the available nitrogen source. However, this type of approach can also generate responses in the yeast that do not necessarily alter the expression of genes related to nitrogen metabolism. In this work, by using intraspecific hybridisation of wild wine yeast strains we obtained genetically and oenologically similar strains with differences in the consumption of nitrogen sources. Using the same must, the global expression patterns of these yeasts were compared by microarrays, the analysis of which identified 276 genes that varied in their expression between the strains analysed. The functional analysis of the genes with a known function indicates that some participate in nitrogen metabolism, alcoholic fermentation, ion transport and transcriptional regulation. Furthermore, differences were observed in the expression of genes which have been partially associated to nitrogen, as in the case of ZRT1 and ATO2. Interestingly, many of the genes identified have no known function or have not been previously associated to this phenotype.


Assuntos
Redes e Vias Metabólicas/genética , Nitrogênio/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Vinho/microbiologia , Cruzamentos Genéticos , Perfilação da Expressão Gênica , Genes Fúngicos , Análise em Microsséries
6.
J Appl Microbiol ; 108(5): 1850-8, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20163487

RESUMO

AIMS: To study genomic and phenotypic changes in wine yeasts produced in short time periods analysing yeast strains possibly derived from commercial strains recently dispersed. METHODS AND RESULTS: We conducted a genomic and phenotypic comparison between the commercial yeast strain EC1118 and two novel strains (LV CB and L-957) isolated from different wine areas industrially intervened <20 years ago. Molecular analysis by amplified fragment length polymorphism (AFLP) and RAPD-PCR was not able to distinguish between these strains. However, comparative genomic hybridization (aCGH) showed discrete DNA gains and losses that allowed unequivocal identification of the strains. Furthermore, analysis of aCGH data supports the hypothesis that strains LV CB and L-957 are derivatives from strain EC1118. Finally, scarce phenotypic differences in physiological and metabolic parameters were found among the strains. CONCLUSION: The wine yeasts have a very dynamic genome that accumulates changes in short time periods. These changes permit the unique genomic identification of the strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This study permits the evaluation of microevolutive events in wine yeasts and its relationship with the phenotype in this species.


Assuntos
Variação Genética , Genoma Fúngico , Fenótipo , Saccharomyces cerevisiae/fisiologia , Vinho/microbiologia , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Hibridização Genômica Comparativa , Dosagem de Genes , Técnica de Amplificação ao Acaso de DNA Polimórfico , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/isolamento & purificação , Proteínas de Saccharomyces cerevisiae/genética
7.
Folia Biol (Praha) ; 56(4): 183-93, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20974051

RESUMO

BCL3 is a putative proto-oncogene deregulated in haematopoieitic and solid tumours. It has been suggested that its oncogenic effects could be mediated, at least in part, by inducing proliferation and inhibiting cell death. To provide more insight into the mediators of these effects, we used an unbiased approach to analyse the mRNA expression changes after knocking-down BCL3 using specific shRNAs. One hundred eighty genes were up-regulated and sixtynine genes were down-regulated after knocking down BCL3. Function analyses showed enrichment in genes associated with cellular growth and proliferation, cell death and gene expression. We found that STAT3, an important oncogene in human cancer, was the central node of one of the most significant networks. We validated STAT3 as a bona fide target of BCL3 by additional interference RNA and in silico analyses of previously reported lymphoma patients.


Assuntos
Regulação Neoplásica da Expressão Gênica , Proteínas Proto-Oncogênicas/metabolismo , Fator de Transcrição STAT3/metabolismo , Fatores de Transcrição/metabolismo , Neoplasias do Colo do Útero/genética , Proteína 3 do Linfoma de Células B , Linhagem Celular Tumoral , Regulação para Baixo , Feminino , Perfilação da Expressão Gênica , Células HeLa , Humanos , Análise de Sequência com Séries de Oligonucleotídeos , Proteínas/genética , Proteínas/metabolismo , Proto-Oncogene Mas , Proteínas Proto-Oncogênicas/genética , Interferência de RNA , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Fator de Transcrição STAT3/genética , Fatores de Transcrição/genética , Regulação para Cima , Neoplasias do Colo do Útero/metabolismo
8.
FEBS Lett ; 371(2): 132-6, 1995 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-7672112

RESUMO

A total of 11 manganese peroxidase isoenzymes (MnP1-MnP11) with isoelectric points (pIs) in the range of 4.58-3.20 were isolated from liquid- and solid-state cultures of the basidiomycete Ceriporiopsis subvermispora. In the presence of hydrogen peroxide, these isoenzymes showed different requirements for Mn(II) in the oxidation of vanillylacetone, o-dianisidine, p-anisidine and ABTS, whereas oxidation of guaiacol by any isoenzyme did not take place when this metal was omitted. Km values for o-dianisidine and p-anisidine in the absence of Mn(II) are in the range of 0.5-1.0 mM and 4.5-42.0 mM, respectively. Oxalate and citrate, but not tartrate, accelerate the oxidation of o-dianisidine, both in the presence and in the absence of Mn(II). MnPs from this fungus are able to oxidize kojic acid without externally added hydrogen peroxide, indicating that they can also act as oxidases. In this reaction, however, the requirement for Mn(II) is absolute.


Assuntos
Basidiomycota/enzimologia , Isoenzimas/metabolismo , Peroxidases/metabolismo , Benzotiazóis , Dianisidina/metabolismo , Eletroforese em Gel de Poliacrilamida , Guaiacol/análogos & derivados , Guaiacol/metabolismo , Ponto Isoelétrico , Isoenzimas/química , Isoenzimas/isolamento & purificação , Manganês/farmacologia , Oxalatos/farmacologia , Ácido Oxálico , Oxirredução , Peroxidases/química , Peroxidases/isolamento & purificação , Pironas/metabolismo , Ácidos Sulfônicos/metabolismo
9.
Drugs Exp Clin Res ; 25(2-3): 133-41, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-10370876

RESUMO

An intervention study was performed to evaluate the influence of a Mediterranean diet, a high fat diet, and their supplementation with red wine in moderate amounts, on biochemical, physiological, and clinical parameters related to atherosclerosis and other chronic diseases. For 3 months two groups of 21 male volunteers each, received either a Mediterranean diet or a high fat diet; during the second month, red wine was added isocalorically, 240 ml/day. Participants were kept under close medical and nutritional surveillance. At days 0, 30, 60 and 90, clinical, physiological and biochemical evaluations were made. Plasma vitamin C was significantly decreased in the high fat diet group compared to the Mediterranean diet group. After wine supplementation to the Mediterranean diet, a significant 13.5% increase in plasma vitamin C was observed. Furthermore, when wine was added vitamin E decreased significantly in plasma, 15% in the high fat diet and 26% in the Mediterranean diet. Total plasma antioxidant capacity (total antioxidant reactivity) increased 28% above basal levels in the Mediterranean diet group, but not in the high fat diet group. In both groups, wine induced a marked increase in total antioxidant reactivity above basal levels, 56% and 23%, respectively. Oxidative DNA damage, detected as 8-hydroxydeoxyguanosine (8-OHdG) levels in blood leukocyte DNA, was markedly increased by the high fat diet; however, it was strongly reduced, to approximately 50% basal values, after wine supplementation, both in the high fat diet and Mediterranean diet groups. Endothelial function, evaluated noninvasively as flow-mediated vascular reactivity of the brachial artery, was suppressed by the high fat diet, and was normal after wine supplementation. These effects are attributed to oxidative stress associated with a high fat diet, and to the elevated plasma antioxidant capacity associated with wine consumption and the Mediterranean diet. The results presented support the following conclusions: a high fat diet induces oxidative stress; a diet rich in fruits and vegetables enhances antioxidant defenses; wine supplementation to a high fat or a Mediterranean diet increases plasma antioxidant capacity, decreases oxidative DNA damage, and normalizes endothelial function.


Assuntos
Antioxidantes/análise , Dano ao DNA/efeitos dos fármacos , Dieta Aterogênica , Gorduras na Dieta , Flavonoides , Fenóis/sangue , Fenóis/metabolismo , Polímeros/metabolismo , Vinho/análise , Adulto , Arteriosclerose/prevenção & controle , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/fisiologia , Humanos , Metabolismo dos Lipídeos , Masculino , Estresse Oxidativo/efeitos dos fármacos , Polifenóis
10.
Autoimmun Rev ; 12(5): 567-74, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23207284

RESUMO

Sjögren's syndrome (SS) is a chronic autoimmune disease of undefined etiology. Patients with this syndrome suffer from severe alterations in both the quality and quantity of saliva and tears, due to impaired function of the relevant exocrine glands. Prevalent symptoms experienced by SS-patients include a persistent dry mouth sensation (xerostomia) and dry eyes (keratoconjunctivitis sicca). Water content of saliva depends of acetylcholine levels, glandular innervation, M3R signaling, calcium tunneling and water release, among other factors. However, unstimulated salivary flow correlates only poorly with symptoms of mouth dryness, raising the question as to which other components of saliva may be involved in mouth dryness experienced by SS-patients? Salivary mucins are glycoproteins characterized by the presence of large oligosaccharide side chains attached to the protein backbone. These molecules are key saliva components that are required to sequester water and thereby moisturize, as well as lubricate the oral mucosa. In the labial salivary glands of SS patients, morphological and functional alterations are detectable that affect the maturation and trafficking of salivary mucins. In this review, we will focus the discussion on these aspects of reduced salivary flow and decreased quality of salivary mucins, since they are likely to be responsible for xerostomia in SS-patients.


Assuntos
Mucinas/deficiência , Mucinas/metabolismo , Saliva/metabolismo , Água/metabolismo , Xerostomia/etiologia , Xerostomia/imunologia , Humanos , Saliva/imunologia , Glândulas Salivares/química , Glândulas Salivares/imunologia , Glândulas Salivares/metabolismo , Xerostomia/metabolismo
11.
Rev Med Chil ; 134(5): 575-80, 2006 May.
Artigo em Espanhol | MEDLINE | ID: mdl-16802049

RESUMO

BACKGROUND: Surgery of the aortic arch is a very complex procedure since it requires protective strategies for the brain, heart and rest of the body. AIM: To communicate our experience in the first 23 total or partial replacements of aortic arch. MATERIAL AND METHODS: Retrospective search in the database of the Cardiovascular Surgery Unit for patients subjected to partial or total replacement of the aortic arch since 1998. RESULTS: Between 1988 and 2002, 23 patients were operated. Seventeen had aortic dissection (10 acute and 7 chronic), five had an atherosclerotic aneurysm and one had a traumatic lesion. Thirteen patients were subjected to a replacement of the arch plus ascending aorta, six to a replacement of the arch plus descending aorta and four to a replacement of the arch, ascending and descending aorta. Seven patients had previous operation of the thoracic aorta. Arterial perfusion was done via the femoral artery, axillary artery or a combination of both. A hypothermic circulatory arrest was induced in 22; it was associated with cerebral retro perfusion alone in 8 patients, antegrade cerebral perfusion in 5; isolated or associated axillary perfusion was used in five patients. In seven, procedures on the aortic or mitral valve, or coronary artery operations were added. Operative mortality was 26%, 3 of the 8 patients operated as an emergency and 3 of 15 elective operations. There was no mortality among those without dissection and of 7 chronic dissections, one died. All patients were followed for an average of 45 months. Two patients required reinterventions on the aorta and one for colon cancer. There was one late death of unknown cause. Postoperative complications were agitation, bleeding and temporary vocal cord dysfunction. CONCLUSIONS: There is a learning curve, where more extensive operations, particularly those done as emergency or for dissections, had an increased operative risk.


Assuntos
Aorta Torácica/cirurgia , Aneurisma da Aorta Torácica/cirurgia , Dissecção Aórtica/cirurgia , Dissecção Aórtica/mortalidade , Aneurisma da Aorta Torácica/mortalidade , Encéfalo/irrigação sanguínea , Parada Circulatória Induzida por Hipotermia Profunda , Circulação Extracorpórea , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Resultado do Tratamento
12.
Appl Environ Microbiol ; 64(1): 68-73, 1998 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16349495

RESUMO

The ligninolytic system of the basidiomycete Ceriporiopsis subvermispora is composed of manganese peroxidase (MnP) and laccase. In this work, the source of extracellular hydrogen peroxide required for MnP activity was investigated. Our attention was focused on the possibility that hydrogen peroxide might be generated by MnP itself through the oxidation of organic acids secreted by the fungus. Both oxalate and glyoxylate were found in the extracellular fluid of C. subvermispora cultures grown in chemically defined media, where MnP is also secreted. The in vivo oxidation of oxalate was measured; CO(2) evolution was monitored after addition of exogenous [C]oxalate to cultures at constant specific activity. In standard cultures, evolution of CO(2) from oxalate was maximal at day 6, although the MnP titers were highest at day 12, the oxalate concentration was maximal (2.5 mM) at day 10, and the glyoxylate concentration was maximal (0.24 mM) at day 5. However, in cultures containing low nitrogen levels, in which the pH is more stable, a better correlation between MnP titers and mineralization of oxalate was observed. Both MnP activity and oxidation of [C]oxalate were negligible in cultures lacking Mn(II). In vitro assays confirmed that Mn(II)-dependent oxidation of [C]oxalate by MnP occurs and that this reaction is stimulated by glyoxylate at the concentrations found in cultures. In addition, both organic acids supported phenol red oxidation by MnP without added hydrogen peroxide, and glyoxylate was more reactive than oxalate in this reaction. Based on these results, a model is proposed for the extracellular production of hydrogen peroxide by C. subvermispora.

13.
Arch Biochem Biophys ; 360(2): 215-22, 1998 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-9851833

RESUMO

The kinetics of Mn3+-oxalate formation and decay were investigated in reactions catalyzed by manganese peroxidase (MnP) from the basiomycete Ceriporiopsis subvermispora in the absence of externally added hydrogen peroxide. A characteristic lag observed in the formation of this complex was shortened by glyoxylate or catalytic amounts of Mn3+ or hydrogen peroxide. MnP titers had a minor effect on this lag and did not influence the decay rate of the complex. In contrast, Mn2+ and oxalate drastically affected maximal concentrations of the Mn3+-oxalate complex formed, the decay of which was accelerated at high Mn2+ levels. The highest concentration of complex was obtained at pH 4.0, whereas an inverse relationship was found between the pH of the reaction and the decay rate of the complex with MnP present. In the absence of MnP, the best fit for the decay kinetics of the complex gave an order of 3/2 at concentrations in the range of 30-100 microM, with a kobs = 0.012 min-1 M-0.5 at pH 4.0. The rate constant increases at lower pH values and decreases at high oxalate concentrations. The physiological relevance of these findings is discussed.


Assuntos
Manganês/metabolismo , Oxalatos/metabolismo , Peroxidases/metabolismo , Polyporales/metabolismo , Catalase/metabolismo , Cátions/metabolismo , Ativação Enzimática/efeitos dos fármacos , Glioxilatos/farmacologia , Meia-Vida , Peróxido de Hidrogênio/metabolismo , Peróxido de Hidrogênio/farmacologia , Concentração de Íons de Hidrogênio , Cinética , Compostos de Manganês/farmacologia , Oxidantes/metabolismo , Oxirredução , Peroxidases/antagonistas & inibidores , Peroxidases/isolamento & purificação , Polyporales/enzimologia , Sulfatos/farmacologia , Superóxido Dismutase/metabolismo
14.
Arch Biochem Biophys ; 366(2): 275-82, 1999 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-10356293

RESUMO

The enzyme oxalate oxidase was identified in mycelial extracts of the basidiomycete Ceriporiopsis subvermispora and thereafter purified to homogeneity. The purification procedure included only three steps: Q-Sepharose chromatography, precipitation at pH 3.0, and phosphocellulose chromatography. The enzyme is a 400-kDa homohexamer, as determined by gel permeation in Sephadex G-200 and SDS-polyacrylamide gel electrophoresis. Isoelectrofocusing revealed a pI of 4.2. Optimal activity was obtained at pH 3.5 and at 45 degrees C. The purified enzyme has Km and kcat values of 0.1 mM and 88 s-1, respectively. It is highly specific for oxalate, although it is inhibited at concentrations of this substrate above 2.5 mM. Hystochemistry studies conducted over mycelium slices showed reactions products in both endocellular and periplasmic associated elements. A possible connection between the intracellular metabolism of oxalate and the extracellular ligninolytic activity of the fungus is proposed.


Assuntos
Oxirredutases/química , Oxirredutases/isolamento & purificação , Proteínas de Plantas/química , Proteínas de Plantas/isolamento & purificação , Polyporaceae/enzimologia , Ativação Enzimática , Histocitoquímica , Microscopia Eletrônica , Oxirredutases/fisiologia , Proteínas de Plantas/fisiologia , Polyporaceae/ultraestrutura , Frações Subcelulares/enzimologia , Frações Subcelulares/ultraestrutura
15.
Exp Cell Res ; 204(2): 378-81, 1993 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-8440334

RESUMO

Analysis of epididymal rat sperm RNA strongly suggested the presence of U snRNAs, especially U1 and U2 snRNA. By Northern blot analysis with radiolabeled oligodeoxynucleotide probes, the presence of U1 and U2 snRNA in rat sperm was confirmed. To precisely localize these RNAs, in situ hybridization with antisense and sense oligo probes labeled with digoxigenin was carried out. The results indicate that U1 as well as U2 snRNA are confined to the sperm nucleus.


Assuntos
Núcleo Celular/química , RNA Nuclear Pequeno/análise , Espermatozoides/química , Animais , Sequência de Bases , Hibridização In Situ , Masculino , Dados de Sequência Molecular , Sondas de Oligonucleotídeos , RNA Nuclear Pequeno/química , Ratos
16.
Rev. méd. Chile ; 134(5): 575-580, mayo 2006. tab
Artigo em Espanhol | LILACS | ID: lil-429863

RESUMO

Background: Surgery of the aortic arch is a very complex procedure since it requires protective strategies for the brain, heart and rest of the body. Aim: To communicate our experience in the first 23 total or partial replacements of aortic arch. Material and methods: Retrospective search in the database of the Cardiovascular Surgery Unit for patients subjected to partial or total replacement of the aortic arch since 1998. Results: Between 1988 and 2002, 23 patients were operated. Seventeen had aortic dissection (10 acute and 7 chronic), five had an atherosclerotic aneurysm and one had a traumatic lesion. Thirteen patients were subjected to a replacement of the arch plus ascending aorta, six to a replacement of the arch plus descending aorta and four to a replacement of the arch, ascending and descending aorta. Seven patients had previous operation of the thoracic aorta. Arterial perfusion was done via the femoral artery, axillary artery or a combination of both. A hypothermic circulatory arrest was induced in 22; it was associated with cerebral retro perfusion alone in 8 patients, antegrade cerebral perfusion in 5; isolated or associated axillary perfusion was used in five patients. In seven, procedures on the aortic or mitral valve, or coronary artery operations were added. Operative mortality was 26%, 3 of the 8 patients operated as an emergency and 3 of 15 elective operations. There was no mortality among those without dissection and of 7 chronic dissections, one died. All patients were followed for an average of 45 months. Two patients required reinterventions on the aorta and one for colon cancer. There was one late death of unknown cause. Postoperative complications were agitation, bleeding and temporary vocal cord dysfunction. Conclusions: There is a learning curve, where more extensive operations, particularly those done as emergency or for dissections, had an increased operative risk.


Assuntos
Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Dissecção Aórtica/cirurgia , Aorta Torácica/cirurgia , Aneurisma da Aorta Torácica/cirurgia , Dissecção Aórtica/mortalidade , Aneurisma da Aorta Torácica/mortalidade , Encéfalo/irrigação sanguínea , Parada Circulatória Induzida por Hipotermia Profunda , Circulação Extracorpórea , Estudos Retrospectivos , Resultado do Tratamento
17.
Rev. chil. anest ; 22(2): 85-94, nov. 1993. tab, ilus
Artigo em Espanhol | LILACS | ID: lil-130989

RESUMO

Los pacientes sometidos a cirugía carotídea tienen un mayor riesgo cardiovascular. Esto y las características de la cirugía hacen deseable una óptima estabilidad hemodinámica en el intra y posoperatorio. El objetivo fue comparar dos técnicas de inducción anestésica en pacientes sometidos a endarterectomía carotídea electiva, en relación a la respuesta hemodinámica durante los períodos: posinducción, posintubación, preincisión y posincisión estandarizados previamente y condiciones de extubación al final de la cirugía. Un total de 21 pacientes fueron aleatoriamente asignados a uno de dos esquemas de inducción. Grupo I (G I)(n=10) Fentanyl 5µgrùkg-1 más Tiopental 4 mgùkg-1 y Grupo II (G II)(n=11) Alfentanil 120 µgrùkg-1. Entre grupos hubo diferencia sólo en los valores de PA, IC y LVSWI en el período posinducción. En ambos grupos hubo un descenso significativo en PA, IC y LVSWI en los períodos preincisión y posincisión. En el G II disminuyó, además, la FC presentándose un caso de bricardia extrema de difícil tratamiento. Dos pacientes, uno de cada grupo, no fueron estubados por no cumplir los criterios prefijados. La inducción con Alfentanil mostró menos variabilidad hemodinámica entre los períodos de medición, aunque la tendencia en ambos grupos fue al descenso de la PA hacia los períodos finales de estudio. La extubación fue comparable


Assuntos
Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Alfentanil/farmacologia , Endarterectomia das Carótidas , Hemodinâmica , Tiopental/farmacologia , Anestésicos/farmacologia , Complicações Intraoperatórias/prevenção & controle , Pressão Sanguínea , Procedimentos Cirúrgicos Vasculares
18.
Rev. chil. cir ; 38(3): 183-7, 1986. ilus
Artigo em Espanhol | LILACS | ID: lil-76997

RESUMO

La reparación paliativa de los aneurismas de la aorta ascendente tiene una recidiva alta por tratarse de una patología degenerativa que compromete la aorta desde el anillo valvular. Actualmente se prefiere el tratamiento definitivo que consiste en el reemplazo de la aorta ascendente junto con la válvula aórtica. Entre marzo de 1978 y octubre de 1983 hemos intervenido 8 pacientes con esta patología. Cinco casos presentaban disección de la aorta de tipo proximal y 2 se operaron en etapa aguda. La indicación operatória fue insuficiencia aórtica grave en 6 pacientes y disección aguda en los 2 restantes. La reparación del aneurisma se realizó con prótesis de dacrón que tiene incorporada una válvula de Bejork-Shiley sobre la cual se reimplantaron los ostia coronarios. El estudio histológico de la aorta demostró necrosis quística de la túnica media en todos los casos. Dos pacientes fallecieron por complicaciones quirúrgicas y los 6 restantes fueron dados de alta bien. El control alejado (x = 21,6 meses) reveló que 5 pacientes están asintomáticos y sin recidiva, 1 falleció a los 15 meses. La técnica de esta grave patología con buenos resultados alejados y sin recidivas


Assuntos
Adulto , Pessoa de Meia-Idade , Humanos , Masculino , Feminino , Aneurisma Aórtico/cirurgia , Prótese Vascular , Próteses Valvulares Cardíacas/métodos
19.
Rev. chil. cardiol ; 12(1): 13-7, ene.-mar. 1993. tab
Artigo em Espanhol | LILACS | ID: lil-125383

RESUMO

Durante los últimos 20 años, las indicaciones y técnica quirúrgica de la revascularización miocárdica han evolucionado en forma importante, por lo cual nos ha interesado estudiar estos cambios y su influencia sobre los resultados obtenidos en pacientes sometidos a este tratamiento quirúrgico. Con este objeto, se analizaron los factores clínicos (edad, capacidad funcional, antecedentes mórbidos, factores de riesgo y patologías asociadas), la cinecoronariografía selectiva, la técnica quirúrgica, y los resultados de la intervención (mortalidad, morbilidad y estadía hospitalaria) de 400 pacientes de un total de 2.410 casos intervenidos entre 1970-1990. Estos pacientes fueron divididos en cuatro grupos que incluyen los primeros cien pacientes operados en forma consecutiva de puente aortocoronario exclusivo (electivos o urgencias) durante los años 1975 (grupo I), 1980 (grupo II), 1985 (grupo III) y 1990 (grupo IV). Los cambios observados en la técnica quirúrgica incluyeron el uso progresivamente mayor de la arteria mamaria interna (20%, 12%, 60%, 86%, respectivamente), el uso de magnificación óptica en el grupo II y siguientes y luego hipotermia a 25§C y cardioplejía usados en los grupos III y IV. La edad promedio, el porcentaje de pacientes mayores de 70 años, de pacientes intervenidos con infarto reciente (<30 días) de enfermos con lesión de tres vasos, y de pacientes que recibieron tres o más puentes aortocoronarios, fueron progresivamente mayores en el tiempo, y comparados los grupos entre sí, la diferencia tuvo significancia estadística (p<0,001). En cambio, la estadía intrahospitalaria postoperatoria fue decreciente durante el período analizado (14, 12, 9 y 9 días, respectivamente) y la morbilidad y mortalidad permanecieron estables, esta última en el 1%. Concluimos que la cirugía de revascularización miocárdica se ha ido efectuando en pacientes progresivamente más graves y con factores de riesgo mayores, manteniendo la morbimortalidad estable


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Revascularização Miocárdica/métodos , Complicações Pós-Operatórias/epidemiologia , Revascularização Miocárdica/tendências , Procedimentos Cirúrgicos Operatórios/tendências
20.
Rev. chil. cir ; 45(3): 225-8, jun. 1993. ilus
Artigo em Espanhol | LILACS | ID: lil-119829

RESUMO

El acceso auricular izquierdo clásico, por detrás y paralelo al surco interauricular, no permite una buena exposición de la válvula mitral cuando la aurícula izquierda es de tamaño normal o sólo ligeramente aumentado. En reoperaciones, este acceso auricular izquierdo requiere de mayor disección del corazón, además, en esta situación, el anillo mitral se fija en una posición de más difícil exposición. Entre diciembre 1991 y octubre 1992, 25 pacientes fueron operados con un nuevo acceso auricular izquierdo transeptal y superior combinado. Se efectuaron 23 procedimientos mitrales (7 reoperaciones), en 8 casos asociados a otros procedimientos quirúrgicos cardiovasculares, una sección de Haz paraespecífico lateral izquierdo y una resección de mixoma auricular izquierdo. La técnica consiste en una amplia auriculotomía derecha, paralela y a 1,5 cm al surco aurículo-ventricular, exponiendo el tabique interauricular totalmente, el que es incidido desde el limbo inferior de la fosa ovalis hasta la incisión auricular derecha previa, extendiendo luego la incisión por el techo de la aurícula izquierda. En todos los casos se obtuvo un excelente acceso a la cavidad auricular izquierda y una muy buena exposición de la válvula mitral. No se produjeron desgarros de la pared auricular izquierda. No hubo hemorragia intraoperatoria por líneas de suturas y ningún paciente fue reexplorado por sangrado postoperatorio. El control ecocardiográfico no demostró comunicación interauricular residual en ningún caso. No hubo trastornos permanentes de la conducción aurículo-ventricular. No se observó una prolongación mayor del tiempo operatorio total, y en reoperaciones se requirió de menor disección del corazón. Este nuevo abordaje auricular izquierdo permite un excelente acceso a la aurícula izquierda, especialmente cuando ésta es de tamaño normal o ligeramente aumentado, y en reoperaciones cardíacas, y por lo tanto una muy buena exposición de la válvula mitral, sin distorsionar ninguna de las estructuras de ésta. La técnica no prolonga el procedimiento operatorio y no tiene morbilidad intrínseca


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Átrios do Coração/cirurgia , Insuficiência da Valva Mitral/cirurgia , Cirurgia Torácica , Próteses Valvulares Cardíacas/métodos
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