ABSTRACT
Desde la llegada de los stents convencionales y farmacoactivos han disminuido considerablemente los eventos de revascularización quirúrgica, sin embargo la trombosis y reestenosis son 2 factores que, aunque han disminuido, permanecen como complicaciones importantes. Existen varios factores que predisponen a la trombosis y a la reestenosis intrastent. La angiografía convencional tiene serias limitaciones para determinar las causas de la falla del stent. La tomografía de coherencia óptica es una técnica sumamente sensible para determinar las causas de trombosis y reestenosis del stent.
Since the advent of bare metal and drug-eluting stents, the surgical revascularization have declined considerably, however the thrombosis and in-stent restenosis are important complications of these devices. There are several factors that predispose to thrombosis and in-stent restenosis. Conventional angiography has serious limitations to determine the causes of stent failure. Optical coherence tomography is a very sensitive technique to determine the cause of thrombosis and in-stent restenosis.
Subject(s)
Humans , Male , Middle Aged , Coronary Restenosis/diagnosis , Coronary Restenosis/surgery , Coronary Thrombosis/diagnosis , Coronary Thrombosis/surgery , Drug-Eluting Stents , Prosthesis Failure , Tomography, Optical Coherence , Coronary Restenosis/etiology , Coronary Thrombosis/etiology , Surgery, Computer-AssistedABSTRACT
Since the advent of bare metal and drug-eluting stents, the surgical revascularization have declined considerably, however the thrombosis and in-stent restenosis are important complications of these devices. There are several factors that predispose to thrombosis and in-stent restenosis. Conventional angiography has serious limitations to determine the causes of stent failure. Optical coherence tomography is a very sensitive technique to determine the cause of thrombosis and in-stent restenosis.
Subject(s)
Coronary Restenosis/diagnosis , Coronary Restenosis/surgery , Coronary Thrombosis/diagnosis , Coronary Thrombosis/surgery , Drug-Eluting Stents , Prosthesis Failure , Tomography, Optical Coherence , Coronary Restenosis/etiology , Coronary Thrombosis/etiology , Humans , Male , Middle Aged , Surgery, Computer-AssistedABSTRACT
Wastewaters generated from the production of ethanol from sugar cane molasses may have detrimental effects on the environment due to their high chemical oxygen demand and dark brown color. The color is mainly associated with the presence of melanoidins, which are highly recalcitrant to biodegradation. We report here the induction of laccases by molasses wastewaters and molasses melanoidins in the basidiomycetous fungus Trametes sp. I-62. The time course of effluent decolorization and laccase activity in the culture supernatant of the fungus were correlated. The expression of laccase genes lcc1 and lcc2 increased as a result of the addition of complete molasses wastewater and its high molecular weight fraction to fungal cultures. This is the first time differential laccase gene expression has been reported to occur upon exposure of fungal cultures to molasses wastewaters and their melanoidins.
Subject(s)
Gene Expression Regulation, Fungal , Laccase/genetics , Laccase/metabolism , Polymers/metabolism , Polyporales/metabolism , Waste Disposal, Fluid , Biodegradation, Environmental , Molasses/analysis , Polyporales/enzymology , Polyporales/genetics , Transcription, GeneticABSTRACT
Nine phenolic compounds (p-coumaric acid, ferulic acid, guaiacol, syringol, p-methoxyphenol, pyrocatechol, phloroglucinol, 3,5-dihydroxybenzoic acid, and syringaldazine) were tested for their ability to increase laccase production in the ligninolytic basidiomycete Trametes sp. I-62. All these compounds resulted in increases in laccase activity, with the highest levels being detected in the presence of p-coumaric acid (273-fold) and guaiacol (73-fold). The three laccase isozyme genes in this fungus lcc1, lcc2, and lcc3 are differentially expressed in the presence of some of these aromatics with total lcc transcript levels differing markedly depending on the aromatic compound tested. Guaiacol (the best inducer of lcc gene transcription) and p-coumaric acid selectively induced expression of lcc1 and lcc2; ferulic acid induced lcc3 expression, while 3,5-dihydroxybenzoic acid had no marked effect on laccase gene transcription. The results demonstrate that close-related aromatic compounds appear to have different effects on both laccase activity levels and lcc gene expression in this basidiomycete.
Subject(s)
Gene Expression Regulation, Fungal/drug effects , Hydrocarbons, Aromatic/pharmacology , Laccase/metabolism , Polyporales/enzymology , Coumaric Acids/pharmacology , Enzyme Activators/pharmacology , Gene Expression , Genes, Fungal , Guaiacol/pharmacology , Hydroxybenzoates/pharmacology , Isoenzymes/genetics , Laccase/biosynthesis , Phenols/pharmacology , Polyporales/genetics , Propionates , RNA, Fungal/analysis , RNA, Fungal/biosynthesis , RNA, Messenger/analysis , RNA, Messenger/biosynthesis , Resorcinols , Transcription, GeneticABSTRACT
The study of isozymes has to date been successfully applied in the identification of a number of different microorganisms, particularly fungi. Trametes sp. I-62 (CECT20197) is a white-rot fungus with potential biotechnological applications. It has been identified as Trametes on the basis of the mycelium morphology, but the features observed are not sufficient for its classification at the species level. Given that this strain produces a very characteristic laccase isozyme pattern, in the present work we compared its isozyme profile with the profiles produced by ten other very closely phylogenetically related fungi from the Polyporaceae family. The data indicates that Trametes sp. I-62 could not be included in any of the taxonomically related species which were analysed in the present work. Extracellular laccases were detected in nine of the fungi studied and in four of them namely Trametes subectypus, Trametes pavonia, Trametes ochracea and Trametes gallica, this is the first report of such activity. Given that phylogenetically related fungi growing in the same culture conditions exhibit different laccase patterns, these isozymes could be used as an additional criterion for fungal identification.
Subject(s)
Laccase/metabolism , Polyporaceae/enzymology , Culture Media , Isoenzymes/metabolism , Polyporaceae/growth & development , Species Specificity , Time FactorsABSTRACT
Laccases produced by white rot fungi are involved in the degradation of lignin and a broad diversity of other natural and synthetic molecules, having a great potential for biotechnological applications. They are frequently encoded by gene families, as in the basidiomycete Trametes sp. strain I-62, from which the lcc1, lcc2, and lcc3 laccase genes have been cloned and sequenced. A multiplex reverse transcription-PCR method to simultaneously study the expression of these genes was developed in this study. The assay proved to be quick, simple, highly sensitive, and reproducible and is particularly valuable when numerous samples are to be analyzed and/or if the amount of initial mRNA is limited. It was used to analyze the effect of 3,4-dimethoxybenzyl alcohol (veratryl alcohol) and two of its isomers (2,5-dimethoxybenzyl alcohol and 3,5-dimethoxybenzyl alcohol) on differential laccase gene expression in Trametes sp. strain I-62. These aromatic compounds produced different induction patterns despite their chemical similarity. We found 2,5-dimethoxybenzyl alcohol to be the best inducer of laccase activity while also producing the highest increase in gene expression; 3,5-dimethoxybenzyl alcohol was the next best inducer. Transcript amounts of each gene fluctuated dramatically in the presence of these three inducers, while the total amounts of laccase mRNAs seemed to be modulated by a coordinated regulation of the different genes.
Subject(s)
Basidiomycota/enzymology , Basidiomycota/growth & development , Gene Expression Regulation, Fungal , Laccase/metabolism , Reverse Transcriptase Polymerase Chain Reaction/methods , Basidiomycota/genetics , Benzyl Alcohols/metabolism , Culture Media/chemistry , Isomerism , Laccase/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolismABSTRACT
The strain Trametes sp. I-62 (CECT 20197) is a white-rot fungus with great potential for biotechnological applications in the fields of industrial waste water decolorization and clean up. Three laccase genes: lcc1, lcc2 and lcc3 have been cloned and sequenced from this basidiomycete. In this work, the coding regions of the corresponding cDNAs have been synthesized, cloned, and sequenced. They are 1563, 1563 and 1575 bp in length, respectively. Former putative intron/exon structures from genomic DNA are fully confirmed by match analysis with our cDNA sequences. Using Polymerase Chain Reaction--Restriction Fragment Length Polymorphism (PCR-RFLP) analysis, an additional laccase cDNA was also identified, corresponding to a new gene, lcc1A, which displayed 99.6% identity with lcc1 at protein level. Such high similarity between lcc1 and lcc1A sequences, and the comparison with reports from other basidiomycete laccases, suggest that in this strain these two genes are allelic variants.
Subject(s)
Basidiomycota/enzymology , DNA, Complementary/genetics , Oxidoreductases/genetics , Sequence Analysis, DNA , Amino Acid Sequence , Base Sequence , Basidiomycota/genetics , Cloning, Molecular , Genome, Fungal , Laccase , Molecular Sequence Data , Oxidoreductases/chemistry , Oxidoreductases/metabolism , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Sequence AlignmentABSTRACT
Pectinesterase was extracted from potato alpha cultivar, purified and partially characterized The used protocol resulted in a 58.8-fold purification (51 850.2 units/mg protein) with 15.5 percent recovery of pectinesterase activity. The purified enzyme had a molecular weight of 27 kDa and its isoelectric point was around 4.5 with pH and temperature optima of 8.0 and 60°C, respectively. The purified enzyme had a single symmetric peak of specific activity after chromatographic steps. The homogeneity of the purified pectinesterase was confirmed by gel filtration and polyacrylamide electrophoresis gel.
A pectinesterase foi extraída da batata (cultivar do alfa), purificada e parcialmente caracterizada. O protocolo usado levou a uma proteína purificada 58,8 vezes (51 850,2 units/mg da proteína) com uma recuperação de 15,5 por cento da atividade da proteína. A enzima purificada apresentou um peso molecular de 27 kDa e seu ponto isoelétrico foi ao redor 4,5. A pectinesterase exibiu pH e temperatura ótimos de respectivamente 8,0 e 60°C. A enzima purificada apresentou um único pico simétrico de atividade específica após as etapas de cromatografia. A homogeneidade da pectinesterase purificada foi confirmada por filtração em gel e por eletroforese em gel de poliacrilamida.
ABSTRACT
La asociación de bacteria y hongos en las vulvovaginitis motiva un tratamiento combinado para evitar la recaída de la paciente. En este estudio se incluyeron 36 pacientes con vaginitis o vaginosis y se les practico exámenes bacteriológicos para determinar el organismo causante del cuadro clínico. La recurrencia del padecimiento se relacionó directamente con más de seis infecciones al año (39 por ciento). Los síntomas más severos se relacionaron con virus del papiloma humano (VPH) o Eschericha coli. La combinación de itraconazol/secnidazol fue efectiva tanto clínica como bacteriológicamente. Los casos resistentes al tratamiento fueron por VPH o gérmenes gramnegativos. La combinación es inocua y tuvo leves efectos secundarios sólo en 4.41 por ciento de los casos sin necesidad de suspender el tratamiento
Subject(s)
Humans , Female , Adolescent , Adult , Middle Aged , Drug Therapy, Combination , Itraconazole/administration & dosage , Vaginitis/drug therapy , Vaginitis/microbiology , Antifungal Agents/administration & dosage , Antiparasitic Agents , Vaginal SmearsABSTRACT
El principal síntoma de vaginitis y vaginosis es la leucorrea. En el presente estudio se valoró la eficacia y seguridad de la combinación de secnidazol e itraconazol en dos poblaciones de mujeres: 48 del área rural y 51 de la urbana. Se observó que la administración de dos cápsulas de la combinación de 166.6 mg de secnidazol y 33.3 mg de itraconazol cada 12 horas por tres dias fue eficaz tanto en medio urbano como rural. Ninguna de las pacientes suspendió el tratamiento por efectos colaterales, ya que sólo se reportaron como leves 6.7 por ciento de los casos en ambas poblaciones
