Browsing by Author "Castillo, Luis"

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    Cell wall modifications in Saccharomyces cerevisiae wine yeast through adaptive laboratory evolution with Tebuconazole
    (2025) Becerra Bevensee, Kevin Andrew; Plaza, Verónica; Castillo, Luis; Godoy Olivares, Liliana
    Saccharomyces cerevisiae is the most used yeast for wine production around the world. Several characteristics make this yeast the wine yeast of excellence; among them is a great tolerance to higher concentrations of sugar and alcohol. Nevertheless, some compounds could have detrimental effects on its development like pesticides. Tebuconazole is one of the most common fungicides used in agriculture, belonging to the largest group of fungicides, the triazoles chemical group, that act on sterol biosynthesis. Yeasts have different responses to compensate for stress, and changes in their cell wall are one of the main ones. This work aimed to obtain new variants of S. cerevisiae through adaptive laboratory evolution (ALE) using Tebuconazole as selection pressure and to evaluate changes in yeast cell wall structure, composition, and fermentative behavior. Three new variants of S. cerevisiae were obtained. Analysis of the relative expression of genes associated with cell wall components showed that the third variant obtained YCPUC209C, had overexpression of genes FKS1, FKS3, CHS3, and SED1 in comparison with the original strain, also morphological analysis through TEM microscopy showed that YCPUC209C had an increase of 22.2% of cell wall thickness and 19% increase in the amount of glucan in comparison to the original strain. These cell wall changes were accompanied by hypersensitive to β-1,3-glucanase activity. Increased tolerance to pesticides Buprofezin and Spirotetramat presence during alcoholic fermentation was achieved by YCPUC209C, improving fermentative efficiency parameter. Changes in cell wall structure and composition reported in this work open new lines of analysis like the evaluation of yeast pesticide dissipation capacity since it is reported that components, such as glucan and chitin, can bond to these contaminants, reducing their residues in the wine.
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    bcpmr1 encodes a P-type Ca2+/Mn2+-ATPase mediating cell-wall integrity and virulence in the phytopathogen Botrytis cinerea
    (2015) Plaza, Veronica; Laguees, Yanssuy; Carvajal, Mauro; Perez-Garcia, Luis A.; Mora-Montes, Hector M.; Canessa, Paulo; Larrondo, Luis F.; Castillo, Luis
    The cell wall of fungi is generally composed of an inner skeletal layer consisting of various polysaccharides surrounded by a layer of glycoproteins. These usually contain both N- and O-linked oligosaccharides, coupled to the proteins by stepwise addition of mannose residues by mannosyltransferases in the endoplasmic reticulum and the Golgi apparatus. In yeast, an essential luminal cofactor for these mannosyltransferases is Mn2+ provided by the Ca2+/Mn2+-ATPase known as Pmr1. In this study, we have identified and characterized the Botrytis cinerea pmr1 gene, the closest homolog of yeast PMR1. We hypothesized that bcpmr1 also encodes a Ca2+/Mn2+-ATPase that plays an important role in the protein glycosylation pathway. Phenotypic analysis showed that bcpmr1 null mutants displayed a significant reduction in conidial production, radial growth and diameter of sclerotia. Significant alterations in hyphal cell wall composition were observed including a 83% decrease of mannan levels and an increase in the amount of chitin and glucan. These changes were accompanied by a hypersensitivity to cell wall-perturbing agents such as Calcofluor white, Congo red and zymolyase. Importantly, the Delta bcpmr1 mutant showed reduced virulence in tomato (leafs and fruits) and apple (fruits) and reduced biofilm formation. Together, our results highlight the importance of bcpmr1 for protein glycosylation, cell wall structure and virulence of B. cinerea. (C) 2015 Elsevier Inc. All rights reserved.
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    Intraabdominal hypertension in patients with septic shock
    (Southeastern Surgical Congress, 2007) Regueira, Tomás; Hasbún, Pablo; Rebolledo Acevedo, Rolando Arturo; Galindo, José; Aguirre, Marcia; Romero, Carlos; Castillo, Luis; Bugedo Tarraza, Guillermo; Hernández P., Glenn
    Intraabdominal hypertension (IAH) develops frequently in patients with septic shock. Even a moderate increase in intraabdominal pressure (IAP) in this setting could be associated with high lactate levels. The authors conducted a prospective, observational, nonrandomized control trial in the surgical intensive care unit of an academic tertiary center. Twenty-seven patients with septic shock (septic shock group), and 19 patients undergoing abdominal surgery with more than two risk factors for IAH (postoperative control group) were admitted consecutively to the intensive care unit. IAP was measured every 6 hours during the first 48 hours. IAH was diagnosed with two consecutive measurements greater than 20 mm Hg. The main outcome measures were prevalence of IAH in septic shock and control groups; and comparative lactate levels, norepinephrine requirements and organ dysfunctions in patients with and without IAH in both groups. Fifty-one per cent of patients with septic shock and 31 per cent of control patients developed IAH. Patients with septic shock with and without IAH were comparable in peak norepinephrine dose, sequential organ failure assessment score, and mortality. However, peak lactate levels were significantly higher in patients with septic shock and IAH compared with those without IAH (3.5 mmol/L versus 1.9 mmol/L, P < 0.04). There was a significant positive temporal correlation between IAP and lactate levels in patients with septic shock with IAH. Peak levels of both occurred early and decreased progressively over time. Control patients with and without IAH exhibited comparable peak lactate levels. Intraabdominal hypertension is very common in septic shock and appears to be related to high lactate levels, which diminish as IAP decreases. Future studies should address the usefulness of IAP monitoring in patients with septic shock.
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    Recent Advances in the Study of the Plant Pathogenic Fungus Botrytis cinerea and its Interaction with the Environment
    (2017) Castillo, Luis; Plaza, Verónica; Larrondo Castro, Luis Fernando; Canessa, Paulo
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    Splanchnic ischemia and gut permeability after acute brain injury secondary to intracranial hemorrhage
    (2007) Hernandez, Glenn; Hasbun, Pablo; Velasco, Nicolas; Wainstein, Carol; Bugedo, Guillermo; Bruhn, Alejandro; Klaassen, Julieta; Castillo, Luis
    Introduction Splanchnic ischemia (SI) and increased gut permeability (GP) have been described in acute brain injury (ABI), although their incidence and relation to the type and severity of injury are uncertain. The aim of this study was to evaluate the incidence of both abnormalities in a series of patients with severe ABI secondary to intracranial hemorrhage (ICH) managed with a resuscitation protocol pursuing adequate cerebral and systemic hemodynamics.
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    Step-Wise Ethanol Adaptation Drives Cell-Wall Remodeling and ROM2/KNR4 Activation in Brettanomyces bruxellensis
    (2025) Hernández-Cabello, Leslie; Rojas-Torres, Nachla; Godoy Olivares, Liliana; G-Poblete, Camila; Concha, Yarabi; Plaza, Verónica; Castillo, Luis; Mora-Montes, Héctor M.; Ganga, María Angélica
    Brettanomyces bruxellensis has been described as the main spoilage microorganism in wines due to its ability to produce volatile phenols, which negatively impact the final product’s organoleptic properties. This yeast can grow and survive in environments that are too nutritionally poor and stressful for other microorganisms, and one of the stressful conditions it can endure is the high alcohol content in wine. In this study, cell wall morphology and the expression of some genes related to its composition were characterized under increasing ethanol concentrations to establish a possible ethanol resistance mechanism. B. bruxellensis LAMAP2480 showed greater resistance to β-1,3-glucanase activity when grown in media supplemented with 5% or 10% ethanol compared with the control assay (without ethanol). Transmission electron microscopy showed no significant differences in cell wall thickness during the different adaptation stages. However, the amount of wall polysaccharides and chitin briefly increased at 1% ethanol but returned to baseline at 5% and 10%. The amount of wall-associated protein increased progressively with each increment in ethanol concentration. In addition, overexpression of the ROM2 and KNR4/SMI1 genes was observed at 10% ethanol. These results suggest that the integrity of the cell wall might play an important role in the adaptation of B. bruxellensis to an ethanol-containing medium.