Production of bioethanol from sweet sorghum [Sorghum bicolor L] juice using yeast isolated from fermented sweet sorghum juice / Producción de bioetanol a partir de jugo de sorgo dulce [Sorghum bicolor L] utilizando levadura aislada de jugo de sorgo dulce fermentado

As a sugar-rich plant with no impact on global warming and food security, sweet sorghum can be exploited as an alternative source of renewable bioenergy. This study aimed to examine the potential of sweet sorghum juice for the generation of bioethanol using yeast isolated from the juice. The °Brix of sweet sorghum juice was measured using a digital refractometer. Additionally, 18 wild yeasts isolated from fermented sweet sorghum juice were subjected to various biochemical tests to describe them to identify potential yeast for ethanol production. The morphological and biochemical analyses of the yeasts revealed that all of the yeast isolates were most likely members of the genus Saccharomyces. The most ethanol-tolerant yeast isolate SJU14 was employed for sweet sorghum juice fermentation. A completely randomized factorial design was used with various fermentation parameters, primarily pH, temperature, and incubation period. Then ethanol content was determined using a potassium dichromate solution. According to the ANOVA, the highest ethanol content (18.765%) was produced at 30/26 °C, pH 4.5, and incubated for 96 h. Sweet sorghum juice was found to be an excellent source of potent yeasts, which have important industrial properties like the capacity to grow at high ethanol and glucose concentrations. Moreover, it can be utilized as a substitute substrate for the manufacturing of bioethanol production to lessen the environmental threat posed by fossil fuels. Further research is, therefore, recommended to develop strategically valuable applications of sweet sorghum for enhancing the food system and mitigating climate change.(AU)

Succinic acid production by wine yeasts and the influence of GABA and glutamic acid / Producción de ácido succínico por levaduras enológicas y la influencia del GABA y el ácido glutámico

As a consequence of alcoholic fermentation (AF) in wine, several compounds are released by yeasts, and some of them are linked to the general quality and mouthfeel perceptions in wine. However, others, such as succinic acid, act as inhibitors, mainly of malolactic fermentation. Succinic acid is produced by non-Saccharomyces and Saccharomyces yeasts during the initial stages of AF, and the presence of some amino acids such as γ-aminobutyric acid (GABA) and glutamic acid can increase the concentration of succinic acid. However, the influence of these amino acids on succinic acid production has been studied very little to date. In this work, we studied the production of succinic acid by different strains of non-Saccharomyces and Saccharomyces yeasts during AF in synthetic must, and the influence of the addition of GABA or glutamic acid or a combination of both. The results showed that succinic acid can be produced by non-Saccharomyces yeasts with values in the range of 0.2–0.4 g/L. Moreover, the addition of GABA or glutamic acid can increase the concentration of succinic acid produced by some strains to almost 100 mg/L more than the control, while other strains produce less. Consequently, higher succinic acid production by non-Saccharomyces yeast in coinoculated fermentations with S. cerevisiae strains could represent a risk of inhibiting Oenococcus oeni and therefore the MLF.(AU)

Sonoprocessing enhances the stabilization of fisetin by encapsulation in Saccharomyces cerevisiae cells / El sonoprocesamiento mejora la estabilización de fisetina mediante encapsulación en células de Saccharomyces cerevisiae

The objective of this study was to investigate for the first time the role of S. cerevisiae natural barriers and endogenous cytoplasmatic bodies on the stabilization of fisetin encapsulated via sonoprocessing coupled to freeze-drying (FD) or spray drying (SD). Both protocols of encapsulation improved the resistance of fisetin against thermal treatments (between 60 and 150 °C) and photochemical-induced deterioration (light exposition for 60 days) compared to non-encapsulated fisetin (antioxidant activity retention of approximately 55% and 90%, respectively). When stored under constant relative humidity (from 32.8 to 90%) for 60 days, yeast carriers improved the half-life time of fisetin by up to 4-fold. Spray dried particles were smaller (4.9 μm) and showed higher fisetin release after simulated gastrointestinal digestion (55.7%) when compared to FD. Freeze-dried particles, in turn, tended to agglomerate more than SD (zeta potential −19.7 mV), resulting in reduced loading features (6.3 mg/g) and less efficient protection of fisetin to heat, photo, and moisture-induced deterioration. Overall, spray-dried sonoprocessed fisetin capsules are an efficient way to preserve fisetin against harsh conditions. Altogether, this report shows that sonoprocessing coupled to drying is an efficient, creative, and straightforward route to protect and deliver lipophilic fisetin using yeast capsules for food applications.(AU)

Exogenous glucosylglycerol and proline extend the chronological lifespan of Rhodosporidium toruloides / El glucosilglicerol y la prolina exógenos extienden la vida útil cronológica de Rhodosporidium toruloides

Glucosylglycerol (GG) is an osmolyte found in a few bacteria (e.g., cyanobacteria) and plants grown in harsh environments. GG protects microbes and plants from salinity and desiccation stress. In the industry, GG is synthesized from a combination of ADP-glucose and glycerol-3-phosphate in a condensation reaction catalyzed by glucosylglycerol phosphate synthase. Proline, on the other hand, is an amino acid-based osmolyte that plays a key role in cellular reprograming. It functions as a protectant and a scavenger of reactive oxygen species. Studies on lifespan extension have focused on the use of Saccharomyces cerevisiae. Rhodosporidium toruloides, also known as Rhodotorula toruloides, is a basidiomycetous oleaginous yeast known to accumulate lipids to more than 70% of its dry cell weight. The oleaginous red yeast (R. toruloides) has not been intensely studied in the lifespan domain. We designed this work to investigate how GG and proline promote the longevity of this red yeast strain. The results obtained in our study confirmed that these molecules increased R. toruloides’ viability, survival percentage, and lifespan upon supplementation. GG exerts the most promising effects at a relatively high concentration (100 mM), while proline functions best at a low level (2 mM). Elucidation of the processes underlying these favorable responses revealed that GG promotes the yeast chronological lifespan (CLS) through increased catalase activity, modulation of the culture medium pH, a rise in ATP, and an increase in reactive oxygen species (ROS) accumulation (mitohormesis). It is critical to understand the mechanisms of these geroprotector molecules, particularly GG, and the proclivity of its lifespan application; this will aid in offering clarity on its potential application in aging research.(AU)

Pulsed magnetic field increases the effect of ultraviolet C radiation and thermal shock in aged yeasts / El campo magnético pulsado aumenta el efecto de la radiación ultravioleta C y el choque térmico en levaduras envejecidas

The study of the effects of the magnetic field (MF) on living matter continues to be a dilemma. Until now, the interaction mechanisms of MF with living matter that explain the observed phenomena are unknown. Despite the existing literature and the multiple effects described to date, there are few published articles that study the combined effect of MF with other physical agents during the cellular aging process. In this sense, the aim of this work is to study whether low frequency and intensity pulsed and sinusoidal MF exposure produce alterations in the cell killing effect of ultraviolet C (UVC) radiation and thermal shock during the chronological aging of S. cerevisiae. Yeast cells were exposed to 2.45 mT (50 Hz) sinusoidal MF and 1.5 mT (25 Hz) pulsed MF, during 40 days of aging, in combination with UVC radiation (50 J/m2) and/or thermal shock (52°C). Cell survival was evaluated by clonogenic assay. The exposure of yeast to pulsed MF produces an acceleration of aging, which is not observed in cells exposed to sinusoidal MF. The pulsed MF modifies the cellular response to damaging agents only in aged S. cerevisiae cells. In this sense, the pulsed MF applied increases the damage induced by UVC radiation and by thermal shock. In contrast, the sinusoidal MF used has no effect.(AU)

Tec1 and Ste12 transcription factors play a role in adaptation to low pH stress and biofilm formation in the human opportunistic fungal pathogen Candida glabrata / Los factores de transcripción de Tec1 y Ste12 juegan un papel en la adaptación al estrés de pH bajo y la formación de biopelículas en el patógeno fúngico oportunista humano Candida glabrata

Eukaryotic cells respond to environmental cues through mitogen activated protein kinase (MAPK) signaling pathways. Each MAPK cascade is specific to particular stimuli and mediates specialized responses through activation of transcription factors. In the budding yeast, Saccharomyces cerevisiae, the pheromone-induced mating pathway and the starvation-responsive invasive growth/filamentation pathway generate their distinct outputs through the transcription factors Ste12 and Tec1, respectively. In this study, we report the functional characterization of these transcription factors in the closely related human opportunistic pathogenic yeast Candida glabrata. Two homologues each for S. cerevisiae TEC1 and STE12 were identified in C. glabrata. Both C. glabrata Tec1 proteins contain the N-terminal TEA DNA-binding domain characteristic of the TEA/ATTS transcription factor family. Similarly, the DNA-binding homeodomain shared by members of the highly conserved fungal Ste12 transcription factor family is present in N-terminus of both C. glabrata Ste12 transcription factors. We show that both C. glabrata STE12 genes are at least partial functional orthologues of S. cerevisiae STE12 as they can rescue the mating defect of haploid S. cerevisiae ste12 null mutant. Knockout of one of the STE12 genes (ORF CAGL0H02145g) leads to decreased biofilm development; a stronger biofilm-impaired phenotype results from loss of both CgSTE12 genes in the double deletion mutant (Cgste12ΔΔ). The transcript levels of one of the TEC1 genes (ORF CAGL0M01716g) were found to be upregulated upon exposure to low pH; its deletion causes slightly increased sensitivity to higher concentrations of acetic acid. Heat shock leads to increase in mRNA levels of one of the STE12 genes (ORF CAGL0M01254g).(AU)

Schizosaccharomyces pombe Fzo1 is subjected to the ubiquitin–proteasome-mediated degradation during the stationary phase / Schizosaccharomyces pombe Fzo1 está sujeto a la degradación mediada por ubiquitina-proteasoma durante la fase estacionaria

Mitochondria are highly dynamic organelles that undergo fission and fusion to adapt to the metabolic needs of the cell. Mitofusins are dynamin-like GTPases that play a key role in the regulation of mitochondrial fusion and metabolism. In Saccharomyces cerevisiae, mitofusin Fzo1 levels are controlled by post-translational ubiquitination and degradation. However, it is not clear whether the levels of the Schizosaccharomyces pombe mitofusin Fzo1 are similarly regulated. In this study, we examined the expression S. pombe Fzo1 during normal growth. We showed that Fzo1 protein levels but not mRNA expression levels were reduced during the stationary phase. The protein was stabilized by the proteasome inhibitor bortezomib. Disruption of ubc8 encoding a ubiquitin-conjugating enzyme and rsv2 encoding an S. pombe homolog of S. cerevisiae RPN4 known for activating the expression of genes required for proteasomal biogenesis suppresses the proteasomal degradation of Fzo1 during the stationary phase. Overexpression of fzo1 prevents its degradation. Our results suggest that like S. pombe Fzo1 expression is not regulated by transcription but rather by proteolytic degradation during the stationary phase. Our findings also suggest that although S. cerevisiae and S. pombe Fzo1 proteins are regulated by ubiquitin-proteasomal degradation, different ubiquitin-conjugating enzymes (E2) and ubiquitin ligases (E3) are involved in their degradation.(AU)

Association of Anti-Saccharomyces cerevisiae antibodies with clinical phenotype in spondyloarthritis patients / Asociación de anticuerpos anti-Saccharomyces cerevisiae con fenotipo clínico en pacientes con espondiloartritis

Introduction: The association between spondyloarthritis (SpA) and inflammatory bowel disease (IBD) has been shown in many studies. More recently, with the hypothesis that increased gut inflammation is of etiopathogenic importance in the development of SpA, evaluation of anti-Saccharomyces cerevisiae antibodies (ASCA) has gained increasing relevance. Objective: To study the status and frequency of ASCA in SpA patients and the association of these biomarkers with the clinical profile. Methods: An observational study was performed including 231 SpA patients treated with biologic therapy. ASCA IgA and IgG levels were determined by micro-enzyme-linked immunosorbent assay. Results: Our data showed an increase of ASCA IgA positivity among SpA patients. No relationship was found between ASCA status and the demographic aspects, genetic factors or clinical presentation, except for the association with IBD. Conclusion: Our study confirms that ASCA IgA are elevated in SpA patients. Although there was no evidence of association with a particular disease phenotype, the existence of higher ASCA levels sustains a close relationship between gut and SpA.(AU)

Introducción: La asociación entre la espondiloartritis (SpA) y la enfermedad inflamatoria intestinal (EII) se ha demostrado en muchos estudios. Recientemente, con la hipótesis de que el aumento de la inflamación intestinal es de importancia etiopatogénica en el desarrollo de la SpA, la evaluación de los anticuerpos anti-Saccharomyces cerevisiae (ASCA) ha adquirido una relevancia creciente. Objetivo: Estudiar los niveles de los ASCA en pacientes con SpA, y la asociación de estos biomarcadores con el perfil clínico. Métodos: Se realizó un estudio observacional que incluyó a 231 pacientes con SpA tratados con terapia biológica. Los niveles de ASCA IgA e IgG se determinaron por técnica de ensayo de inmunoabsorción ligado a enzimas. Resultados: Nuestros datos mostraron un aumento de la positividad de ASCA IgA entre los pacientes con SpA. No se encontró ninguna relación entre los ASCA y los aspectos demográficos, factores genéticos o presentación clínica, excepto la asociación con la EII. Conclusión: Nuestro estudio confirma que los niveles de ASCA IgA están elevados en pacientes con SpA. Aunque no hubo evidencia de asociación con un fenotipo de enfermedad en particular, la existencia de niveles más altos de ASCA mantiene una relación estrecha entre el intestino y la SpA.(AU)

DNA susceptibility of Saccharomyces cerevisiae to Zeocin depends on the growth phase

The aim of this study was to evaluate the level of Zeocin-induced double-strand breaks (DSBs) in Saccharomyces cerevisiae cells in a different growth phase, using constant-field gel electrophoresis (CFGE). Saccharomyces cerevisiae diploid strain D7ts1 with enhanced cellular permeability was used. The effects of growth phase and treatment time were evaluated based on Zeocin-induced DSBs, measured by CFGE. Survival assay was also applied. No protoplast isolation was necessary for the detection of DSBs in strain D7ts1. Differences in the response of cells depending on the growth phase were obtained. Cells in exponential growth phase had increased DSB levels only after Zeocin treatment with concentrations equal or higher than 200 μgml−1. Increasing treatment time did not result in higher DSB levels. Oppositely, treatment of cells at the beginning of stationary phase with Zeocin concentrations resulted in more than 1.5-fold increase in DSB levels in comparison with those in untreated cells. Increased DSB levels were measured for all the treatment times. A dose-dependent decrease in cell survival was observed after Zeocin treatment with concentrations in the range of lethality LD20-LD50. A strong negative correlation was calculated between the levels of DSBs and cell survival. New information is provided concerning DNA susceptibility depending on the growth phase. DNA susceptibility is higher in cells at the beginning of stationary phase than those in exponential phase. Data presented here illustrate that the optimized by us CFGE protocol is sensitive and could be used successfully for DSB measurement in Saccharomyces cerevisiae strains with enhanced cellular permeability

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Relación de calprotectina fecal y anti saccharomyces antisacaromices con otros marcadores de actividad en pacientes con espondiloartritis / Relationship between fecal calprotectin, anti-Saccharomyces cerevisiae antibodies and other markers of disease activity in patients with spondyloarthritis

OBJETIVO: Valorar la relación entre la calprotectina fecal, ASCA y marcadores de enfermedad en espondiloartritis (Spa). MÉTODOS: Se evaluaron pacientes ≥18años que cumplían criterios ASAS o criterios modificados de Nueva York. Se analizan criterios de actividad, función física, analíticos y datos demográficos. RESULTADOS: Se incluyeron 33 pacientes. Todos excepto uno tenían valores normales de ASCA. Encontramos correlación significativa entre calprotectina y PCR pero no con otros parámetros, ni entre los niveles de calprotectina y la toma de AINE (p = 0,001). Tampoco entre los niveles de PCR y el uso de AINE. Después de retirar los AINE no encontramos diferencias en los niveles de calprotectina (p = 0,9). CONCLUSIÓN: La calprotectina fecal está elevada en pacientes con Spa y se correlaciona positivamente con la PCR. La elevación de la calprotectina fecal no se ve alterada por el uso de AINE. La cantidad de ASCA no cambia y no se correlaciona con ningún parámetro clínico en el estudio poblacional

OBJECTIVE: To assess the relationship between the increase of fecal calprotectin, anti-Saccharomyces cerevisiae antibodies (ASCA) and disease markers in a group of patients with spondyloarthritis. METHODS: We evaluated patients who were at least 18-years-old and met the Assessment in Spondyloarthritis International Society (ASAS) criteria for spondyloarthritis or the New York modified criteria. We analyzed activity criteria, physical function, analytical criteria (human leukocyte antigen [HLA] B27, fecal calprotectin, presence of ASCA, among others) and demographic data. RESULTS: We included 33 patients. All but one patient had normal ASCA values. We found statistical significance in the correlation of calprotectin with C-reactive protein (CRP) but not with other parameters. We also found a relationship between calprotectin levels and nonsteroidal anti-inflammatory drug (NSAID) intake (P=.001). We found no relationship between CRP levels and NSAID use. After discontinuation of NSAIDs for one month, we found no significant differences in calprotectin levels (P=.9). CONCLUSION: Fecal calprotectin is elevated in patients with spondyloarthritis and correlates positively with CRP. Level of fecal calprotectin is not altered by NSAID use. The amount of ASCA present does not change and does not correlate with any clinical parameters in the study population

Improved ethanol productivity and ethanol tolerance through genome shuffling of Saccharomyces cerevisiae and Pichia stipitis

Genome shuffling by recursive protoplast fusion between Saccharomyces cerevisiae and Pichia stipitis also known as Scheffersomyces stipitis resulted in a promising yeast hybrid strain with superior qualities than those of the parental strains in enhancing biofuel production. Our study focused on the substrate utilization, ethanol fermentation, and ethanol tolerance of the hybrids and the parental strains. The parental strain S. cerevisiae is limited to utilize only hexose sugars, and this leads to decrease in the ethanol yield when they are subjected to ethanol production from lignocellulosic biomass which is rich in pentose sugars. To overcome this limitation, we constructed a hybrid yeast strain through genome shuffling which can assimilate all the sugars present in the fermentation medium. After two rounds of recursive protoplast fusion, there was a higher increase in substrate utilization by hybrid SP2-18 compared to parental strain S. cerevisiae. SP2-18 was able to consume 34% of xylose sugar present in the fermentation medium, whereas S. cerevisiae was not able to utilize xylose. Further, the hybrid strain SP2-18 was able to reach an ethanol productivity of 1.03 g L−1 h−1, ethanol yield 0.447 g/g, and ethanol concentration 74.65 g L−1 which was relatively higher than that of the parental strain S. cerevisiae. Furthermore, the hybrid SP2-18 was found to be stable in the production of ethanol. The random amplified polymorphic DNA profile of the yeast hybrid SP2-18 shows the polymorphism between the parental strains indicating the migration of specific sugar metabolizing genes from P. stipitis, while the maximum similarity was with the parent S. cerevisiae

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Antifungal susceptibility of Saccharomyces cerevisiae and therapy in a murine model of disseminated infection / Sensibilidad antifúngica de Saccharomyces cerevisiae en un modelo murino de infección diseminada

Background: The incidence of systemic infections by Saccharomyces cerevisiae has increased in recent years, especially among immunocompromised patients. Amphotericin B, voriconazole or echinocandins have been used with favorable outcome against systemic infections by this fungus. However, clinical experience is limited and no in vivo studies have been conducted. Aims: We evaluated the in vitro activity of nine antifungal compounds against S.cerevisiae and the in vivo efficacy of those three antifungals showing the highest in vitro activity by using a murine model of systemic infection. Methods: Minimal inhibitory concentrations (MICs) were determined by the microdilution method against three strains of S. cerevisiae. After intravenous infection with 5 × 107 CFUs, animals received liposomal amphotericin B (5 mg/kg), voriconazole (25 mg/kg) or anidulafungin (5 mg/kg). Treatment efficacy was assessed by determining of CFUs/g in liver, kidney, brain, lung and spleen. Results: 5-Fluorocytosine was the most in vitro active compound followed by amphotericin B, voriconazole and anidulafungin. The in vivo study showed that liposomal amphotericin B was the most effective drug driving highest fungal clearance. Conclusions: All treatments reduced the fungal load in comparison to the control group, being liposomal amphotericin B the most effective drug followed by anidulafungin and finally voriconazole

Antecedentes: La incidencia de infecciones sistémicas causadas por Saccharomyces cerevisiae ha aumentado en los últimos años, especialmente entre pacientes inmunodeprimidos. A pesar de que la anfotericina B, el voriconazol o las equinocandinas han dado buen resultado en infecciones sistémicas por este hongo, no se han establecido recomendaciones terapéuticas sólidas. Objetivos: Se evaluó la actividad in vitro de nueve antifúngicos frente a S. cerevisiae y la eficacia in vivo de los tres fármacos con mayor actividad in vitro mediante un modelo murino de infección sistémica. Métodos: Se determinaron las concentraciones mínimas inhibitorias (CMIs) frente a tres cepas de S. cerevisiae por el método de microdilución. Después de la inoculación intravenosa con 5 × 107UFC, los ratones fueron tratados con anfotericina B liposomal (5 mg/kg), voriconazol (25 mg/kg) o anidulafungina (5 mg/kg). La eficacia de los tratamientos se estableció basándose en la determinación de UFC/g en hígado, riñón, cerebro, pulmón y bazo. Resultados: La 5-fluorocitosina fue el compuesto más activo in vitro, seguido por la anfotericina B liposomal, el voriconazol y la anidulafungina. En el estudio in vivo, la anfotericina B liposomal fue el fármaco más eficaz en términos de reducción de la carga fúngica y esterilización de los órganos estudiados. Conclusiones: Todos los tratamientos redujeron la carga fúngica en comparación con el grupo control, y la anfotericina B liposomal fue el antifúngico más efectivo, seguido de la anidulafungina y el voriconazol

Infección por Saccharomyces cerevisiae en un individuo inmunocompetente / Saccharomyces cerevisiae infection in an immunocompetent host

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Phosphate: from stardust to eukaryotic cell cycle control

Phosphorus is a pivotal element in all biochemical systems: it serves to store metabolic energy as ATP, it forms the backbone of genetic material such as RNA and DNA, and it separates cells from the environment as phospholipids. In addition to this 'big hits', phosphorus has recently been shown to play an important role in other important processes such as cell cycle regulation. In the present review, we briefly summarize the biological processes in which phosphorus is involved in the yeast Saccharomyces cerevisiae before discussing our latest findings on the role of this element in the regulation of DNA replication in this eukaryotic model organism. We describe both the role of phosphorus in the regulation of G1 progression by means of the Cyclin Dependent Kinase (CDK) Pho85 and the stabilization of the cyclin Cln3, as well as the role of other molecule composed of phosphorus-the polyphosphate-in cell cycle progression, dNTP synthesis, and genome stability. Given the eminent role played by phosphorus in life, we outline the future of phosphorus in the context of one of the main challenges in human health: cancer treatment (AU)

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Vacuolar proteases from Candida glabrata: acid aspartic protease PrA, neutral serine protease PrB and serine carboxypeptidase CpY. The nitrogen source influences their level of expression / Proteasas vacuolares de Candida glabrata: aspartil proteasa ácida PrA, proteasa neutra PrB y carboxipeptidasa CpY. La fuente de nitrógeno influye en sus niveles de expresión

Background. The Saccharomyces cerevisiae vacuole is actively involved in the mechanism of autophagy and is important in homeostasis, degradation, turnover, detoxification and protection under stressful conditions. In contrast, vacuolar proteases have not been fully studied in phylogenetically related Candida glabrata. Aims. The present paper is the first report on proteolytic activity in the C. glabrata vacuole. Methods. Biochemical studies in C. glabrata have highlighted the presence of different kinds of intracellular proteolytic activity: acid aspartyl proteinase (PrA) acts on substrates such as albumin and denatured acid hemoglobin, neutral serine protease (PrB) on collagen-type hide powder azure, and serine carboxypeptidase (CpY) on N-benzoyl-tyr-pNA. Results. Our results showed a subcellular fraction with highly specific enzymatic activity for these three proteases, which allowed to confirm its vacuolar location. Expression analyses were performed in the genes CgPEP4 (CgAPR1), CgPRB1 and CgCPY1 (CgPRC), coding for vacuolar aspartic protease A, neutral protease B and carboxypeptidase Y, respectively. The results show a differential regulation of protease expression depending on the nitrogen source. Conclusions. The proteases encoded by genes CgPEP4, CgPRB1 and CgCPY1 from C. glabrata could participate in the process of autophagy and survival of this opportunistic pathogen (AU)

Antecedentes. La vacuola de Saccharomyces cerevisiae está involucrada activamente en el mecanismo de autofagia, desarrollando una labor importante en la homeostasis, degradación, recambio proteico, desintoxicación y protección de la célula en condiciones de estrés. Por el contrario, las proteasas vacuolares de Candida glabrata aún no han sido estudiadas por completo. Objetivos. El presente trabajo describe por primera vez la actividad proteolítica vacuolar en C. glabrata. Métodos. Los estudios bioquímicos realizados en C. glabrata pusieron de manifiesto la presencia de diferentes actividades proteolíticas: aspartil proteinasa ácida, que actúa sobre sustratos como la albúmina y la hemoglobina ácida desnaturalizada; serín proteasa neutra, con actividad sobre el substrato de tipo colágeno hide powder azure, y serín carboxipeptidasa, que actúa sobre N-benzoil-tyr-pNa. Resultados. La obtención de una fracción subcelular mostró una elevada actividad enzimática específica de las tres proteasas, lo que permitió confirmar su localización vacuolar. Se realizaron análisis de la expresión de los genes CgPEP4 (CgAPR1), CgPRB1 y CgCPY1 (CgPRC1), codificantes de las actividades proteolíticas aspartil proteasa A, proteasa neutra B y carboxipeptidasa Y, respectivamente. Los resultados reflejan una regulación diferencial de la expresión de la proteasa, dependiendo de la fuente de nitrógeno. Conclusiones. Las proteasas codificadas por los genes CgPEP4, CgPRB1 y CgCPY1 podrían participar en el proceso de autofagia y supervivencia de este patógeno oportunista (AU)

Metabolic effects of B-glucans (Saccharomyces cerevisae) per os administration in rats with streptozotocin-induced diabetes / Efectos metabólicos de los β-glucanos (saccharomyces cerevisiae) a través de la administración oral en ratas con diabetes inducida por estreptozotocina

Introduction: beta-glucans (BG) derived from plant tissues are reported to show metabolic effects. In contrast, those fibers isolated from yeast seem to be more related to immune response modulation. Since diabetic individuals are more susceptible to exacerbation of inflammatory signs, the ingestion of fibers that could conjugate both metabolic and immune effects would be of great importance. Objective: we investigated the effect of BG - Saccharomyses cerevisae - ingestion on glycemic and lipoprotein profile of diabetic rats. Design: twenty-four adult Wistar rats were used, distributed into 4 groups in a design of entirely casualized delineation with a 2 x2 factorial model (with and without diabetes; with and without BG). Diabetes Mellitus was induced by an intraperitoneal injection of 80mg/kg of strepzotocin. Thus, animals with fasting glycemia of over 250mg/dl were considered diabetic. Forty-eight hours after induction, the rats received daily doses of 30 mg/kg of BG or saline solution by gavage during 28 days. Results and discussion: the Groups with DM presented a higher glycemic index and lower C peptide levels than the control groups, in addition to lower weight gain and higher ration consumption, water ingestion and urinary volume. Total cholesterol levels (CT), LDL-C + VLDL-C, plasma triacylglycerides (TAG) and alanine aminotransferase (ALT) were also higher in the diabetic animals (p<0.05). No histopathological hepatic alterations were observed in any of the groups. Furthermore, the diabetic animals present increase in villous:crypt ratio (V:C) in the duodenum, without interference of BG. No alterations in the carcass were observed between the groups. Conclusion: it was concluded that the use of BG significantly reduced the glycemic, TAG and ALT levels, showing its therapeutic potential (AU)

Introdución: los beta-glucanos (BG) derivados de tejidos vegetales se ha informado que muestran efectos metabólicos. Por el contrario, esas fibras aisladas de levadura parecen estar más relacionadas con la modulación de la respuesta inmune. Dado que los individuos con diabetes son más susceptibles a la exacerbación de los signos inflamatorios, la ingestión de fibras sí podría conjugar ambos efectos metabólicos e inmunológicos, lo cual sería de gran importancia. Objetivo: el objetivo de este estudio fue investigar los efectos de la ingestión de los BG —Saccharomyses cerevisiae— en el perfil glucémico y la lipoproteína de ratas diabéticas. Metodos: en el diseño de delineación, totalmente precario, fueron utilizadas 24 ratas Wistar macho adultas distribuidas en cuatro grupos, con un modelo factorial 2x2 (con y sin diabetes, con y sin BG). La diabetes mellitus fue inducida por la inyección intraperitoneal de un 80 mg/kg de estrepzotocina. Por lo tanto, los animales con glucemia en ayunas de más de 250 mg/dl fueron considerados diabéticos. Cuarenta y ocho horas después de la inducción, las ratas recibieron dosis diarias de 30 mg/kg de BG o solución salina mediante alimentación forzada durante 28 días. Resultados y discusión: los grupos con DM presentó el mayor índice glucémico y menores niveles de péptido C que los grupos de control, además de reducir el aumento de peso y un mayor consumo de la ración, la ingestión de agua y el volumen urinario. Los niveles de colesterol total (CT), LDL-C + VLDL-C, triacilglicéridos plasmáticos (TAG) y alanina aminotransferasa (ALT) también fueron más altos en los animales diabéticos (p<0,05), y había alteraciones en los niveles de HDL-C. La ingestión de BG redujo las concentraciones de glucosa en sangre (30%), TAG (32%) y ALT (41%) (p<0.05). No se observaron alteraciones hepáticas en ninguno de los grupos. Además, los animales diabéticos presentaron un aumento de la relación cripta:vellosidades (V:C) en el duodeno, sin interferencia de BG. No se observaron alteraciones en la carcasa entre los grupos. Conclusión: se concluyó que el uso de BG redujo significativamente la glucemia, los niveles de TAG Y ALT, mostrando su potencial terapéutico (AU)

Metabolomic charactetization of yeast cells after dehydration stress

In this study, we analyzed the metabolite features of the yeasts Saccharomyces cerevisiae, Naumovia castellii, and Saccharomyces mikatae. The three species are closely related genetically but differ in their tolerance of desiccation stress. Specifically, we determined whether certain metabolites correlated with cell viability after stress imposition. The metabolomic profiles of these strains were compared before cell desiccation and after cell rehydration. In S. mikatae, the presence of lysine or glutamine during rehydration led to a 20% increase in survival whereas during dehydration the levels of both amino acids in this yeast were drastically reduced (AU)

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The mating type-like loci of Candida glabrata / Los loci de apareamiento de Candida glabrata

Candida glabrata, a haploid and opportunistic fungal pathogen that has not known sexual cycle, has conserved the majority of the genes required for mating and cell type identity. The C. glabrata genome contains three mating-type-like loci called MTL1, MTL2 and MTL3. The three loci encode putative transcription factors, a1, α1 and α2 that regulate cell type identity and sexual reproduction in other fungi like the closely related Saccharomyces cerevisiae. MTL1 can contain either a or α information. MTL2, which contains a information and MTL3 with α information, are relatively close to two telomeres. MTL1 and MTL2 are transcriptionally active, while MTL3 is subject to an incomplete silencing nucleated at the telomere that depends on the silencing proteins Sir2, Sir3, Sir4, yKu70/80, Rif1, Rap1 and Sum1. C. glabrata does not seem to maintain cell type identity, as cell type-specific genes are expressed regardless of the type (or even absence) of mating information. These data highlight important differences in the control of mating and cell type identity between the non-pathogenic yeast S. cerevisiae and C. glabrata, which might explain the absence of a sexual cycle in C. glabrata. The fact that C. glabrata has conserved the vast majority of the genes involved in mating might suggest that some of these genes perhaps have been rewired to control other processes important for the survival inside the host as a commensal or as a human pathogen. This manuscript is part of the series of works presented at the "V International Workshop: Molecular genetic approaches to the study of human pathogenic fungi" (Oaxaca, Mexico, 2012) (AU)

Candida glabrata, una levadura patógena haploide y oportunista, que carece de ciclo sexual conocido (asexual), conserva la mayoría de genes ortólogos requeridos en los procesos de apareamiento, esporulación y la identidad del tipo celular. El genoma de C. glabrata contiene 3 loci de apareamiento llamados MTL1, MTL2 y MTL3 que codifican los presuntos factores de transcripción a1, α1 y α2 que controlan la reproducción sexual e identidad celular en otros hongos, como Saccharomyces cerevisiae con el cual tiene una estrecha relación filogenética. MTL1 puede contener información a o α; MTL2 contiene información a, y MTL3 que contiene información α1 y α2 son loci próximos a 2 telómeros. MTL1 y MTL2 son activos transcripcionalmente mientras que MTL3 está sujeto a un silenciamiento que no es completo, que proviene del telómero y depende de las proteínas Sir2, Sir3, Sir4, yKu70/80, Rif1, Rap1 y Sum1. C. glabrata parece no mantener identidad de tipo celular ya que varios genes específicos de un tipo celular se expresan en todas las células con independencia del tipo de información de apareamiento en los loci MTL, o incluso, en su ausencia. Estos datos ilustran varias diferencias importantes entre la levadura no patógena S. cerevisiae y C. glabrata que podrían explicar la característica asexual en esta última. El hecho de que en C. glabrata se hayan conservado los genes necesarios para el apareamiento podría indicar que es posible que algunos de estos genes se hayan «reorganizado» para controlar otros procesos importantes en la supervivencia de C. glabrata en su huésped, como comensal o como patógeno.Este artículo forma parte de una serie de estudios presentados en el «V International Workshop: Molecular genetic approaches to the study of human pathogenic fungi» (Oaxaca, México, 2012) (AU)

A polyphasic approach to study the dynamics of microbial population of an organic wheat sourdough during its conversion to gluten-free sourdough

To develop a method for organic gluten-free (GF) sourdough bread production, a long-term and original wheat sourdough was refreshed with GF flours. The dynamics of the sourdough microbiota during five months of back-slopping were analyzed by classical enumeration and molecular methods, including PCR-temporal temperature gel electrophoresis (PCR-TTGE), multiplex PCR, and pulsed field gel electrophoresis (PFGE). The results showed that the yeast counts remained constant, although Saccharomyces cerevisiae, present in the initial wheat sourdough, was no longer detected in the GF sourdough, while lactic acid bacteria (LAB) counts increased consistently. In the first phase, which was aimed at obtaining a GF sourdough from wheat sourdough, Lactobacillus sanfranciscensis, L. plantarum, and L. spicheri were the main LAB species detected. During the second phase, aimed at maintaining the GF sourdough, the L. plantarum and L. spicheri populations decreased whereas L. sanfranciscensis persisted and L. sakei became the predominant species. Multiplex PCRs also revealed the presence of several L. sakei strains in the GF sourdough. In a search for the origin of the LAB species, PCR-TTGE was performed on the flour samples but only L. sanfranciscensis was detected, suggesting a flour origin for this typical sourdough species. Thus, while replacement of the wheat flour by GF flour influenced the sourdough microbiota, some of the original sourdough LAB and yeast species remained in the GF sourdough (AU)

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