Shaping the curve from the microscopic transsphenoidal to the endoscopic endonasal approach for the sellar region.

OBJECTIVE: This study aimed to investigate the limitations, barriers, and complications in the early transition from the microscopic transsphenoidal approach (MTA) to the endonasal endoscopic approach (EEA) to the skull base in our institution. METHODS: Technical challenges, as well as clinical features and complications, were compared between MTA, EEA, and mixed cases during the early surgical curve. RESULTS: The period from the early learning curve was 1 year until the EEA protocol was used routinely. A total of 34 patients registered a resection using a transsphenoidal approach. Eighteen patients underwent EEA, 11 underwent MTA, and five underwent a mixed endonasal and microscopic approach. Non-significant differences were found in endocrine outcomes between the three groups. Patients with unchanged or improved visual function were higher in the EEA group (p = 0.147). Non-significant differences were found in terms of the extent of resection (EOR) between groups (p = 0.369). Only 1 (2.9%) patient in the whole series developed a post-operative CSF leaking that resolved with medical management, belonging to the EEA group (5.5%). CONCLUSIONS: The early phase of the learning curve did not affect our series significantly in terms of the EOR, endocrine status, and visual outcomes.

OBJETIVO: Investigar las limitaciones, las barreras y las complicaciones en la transición del abordaje transesfenoidal microscópico (ATM) al abordaje endonasal endoscópico (AEE) para la base del cráneo en nuestra institución. MÉTODO: Se compararon las características clínicas y las complicaciones entre ATM, AEE y casos mixtos durante la curva quirúrgica temprana. RESULTADOS: El periodo desde la curva de aprendizaje inicial fue de 1 año hasta que se utilizó el protocolo AEE de forma sistemática. Un total de 34 pacientes tuvieron una resección por vía transesfenoidal. A 18 pacientes se les realizó AEE, a 11 ATM y a 5 abordaje mixto endonasal y microscópico. Se encontraron diferencias no significativas en los resultados endocrinos entre los tres grupos. Los pacientes con función visual sin cambios o mejorada fueron más en el grupo AEE (p = 0.147). No se encontraron diferencias significativas respecto a la extensión de la resección (p = 0.369). Solo 1 (2.9%) paciente desarrolló una fístula de líquido cefalorraquídeo que se resolvió con manejo médico, perteneciente al grupo AEE (5.5%). CONCLUSIONES: La fase inicial de la curva de aprendizaje no afectó significativamente a nuestra serie en términos de extensión de la resección, estado endocrino y resultados visuales.

Sourdough Yeast Strains Exhibit Thermal Tolerance, High Fermentative Performance, and a Distinctive Aromatic Profile in Beer Wort.

The increasing popularity of home brewing and the fast evolution of craft beer companies have fuelled the interest in novel yeasts as the main actors diversifying the beer portfolio. Here, we have characterized the thermal tolerance and brewing-related features of two sourdough (SD) isolates of Saccharomyces cerevisiae, SDy01 and SDy02, at different temperatures, 20 and 37 °C, comparing them with commercial brew strains, AaB and kNB. The SD strains exhibited tolerance to the main brewing-related stress conditions and increased growth rates and lower lag phases than the reference beer strains at both temperatures. Consistent with this, SDy01 and SDy02 displayed higher fermentative activity in terms of sugar rate depletion and the release of metabolic by-products. Moreover, SDy01 and SDy02 brewing at 20 °C increased their total amount of volatile compounds (VOCs), in particular, their esters and carboxyl compounds, as compared to the reference AaB strain. In contrast, fermentation at 37 °C resulted in a drastic reduction in the number of VOCs in wort fermented with SD yeast, especially in its level of esters. In conclusion, our results stress the high fermentative performance of SD strains in beer wort and their ability to provide a complex and specific aromatic profile at a wide range of temperatures.

Bioprospecting of sourdough microbial species from artisan bakeries in the city of Valencia.

This work describes the characterization of an artisanal sourdough set of bakeries located in the city of Valencia. Culture-dependent and -independent analyses detected Fructilactobacillus sanfranciscensis, Saccharomyces cerevisiae and Kazachstania humilis as dominant species. Nevertheless, specific technological parameters, including backslopping temperature, dough yield, or the addition of salt affected microbial counting, LAB/Yeast ratio, and gassing performance, favouring the appearance of several species of Lactobacillus sp., Limosilactobacillus pontis or Torulaspora delbrueckii as additional players. Sourdough leavening activity was affected positively by yeast counts and negatively by the presence of salt. In addition, the predominance of a particular yeast species appeared to impact the dynamics of CO2 release. Seven important flavour-active compounds (ethyl acetate, 1-hexanol, 2-penthylfuran, 3-ethyl-2-methyl-1,3-hexadiene, 2-octen-1-ol, nonanal and 1-nonanol) were detected in all samples and together with 3-methyl butanol and hexyl acetate represented more than the 53% of volatile abundancy in nine of the ten sourdoughs analysed. Even so, the specific microbial composition of each sample influenced the volatile profile. For example, the occurrence of K. humilis or S. cerevisiae as dominant yeast influenced the composition of major alcohol species, while F. sanfranciscensis and L. pontis positively correlated with aldehydes and octanoic acid content. In addition, relevant correlations could be also found among different technological parameters and between these, volatile compounds and microbial species. Overall, our study emphasises on how differences in technological parameters generate biodiversity in a relatively small set of artisan sourdoughs providing opportunities for excellence and quality baking products.

Balancing Act: A Comprehensive Review of Vestibular Evaluation in Cochlear Implants.

Cochlear implantation, a transformative intervention for individuals with profound hearing loss, has evolved significantly over the years. However, its impact on the vestibular system, responsible for balance and spatial orientation, remains a subject of ongoing research and clinical consideration. This narrative review highlights key aspects of vestibular evaluation in patients undergoing cochlear implantation. Preoperative vestibular assessment is crucial to establish baseline vestibular function and identify any pre-existing balance issues. Various tests, including caloric, rotational chair, vestibular-evoked myogenic potential, and video head impulse tests, play a vital role in evaluating vestibular function. The goal is to assess the risk of vestibular disturbances arising from the surgery, guide surgical planning, and detect pre-existing alterations that could be totally or partially compensated. While some patients experience minimal vestibular disruptions, others may encounter transient or persistent balance issues following cochlear implant surgery. Postoperative vestibular testing allows for the early detection of such disturbances, enabling timely interventions like vestibular rehabilitation and evaluating changes produced due to surgical complications or changes in the patient's prior conditions. Challenges in vestibular evaluation include individual variability in patient responses, the proximity of the cochlea to the vestibular system, and the need to tailor testing protocols to individual needs. Further research is essential to refine testing protocols, minimize vestibular disturbances, and improve outcomes for cochlear implant candidates. A multidisciplinary approach involving otolaryngologists, audiologists, and physical therapists is integral to comprehensive patient care in this context. In conclusion, vestibular evaluation in patients undergoing cochlear implantation is critical for optimizing surgical planning, managing postoperative issues, and enhancing the overall quality of life for individuals embarking on the journey of restored hearing.

Adaptive laboratory evolution for acetic acid-tolerance matches sourdough challenges with yeast phenotypes.

Acetic acid tolerance of Saccharomyces cerevisiae is an important trait in sourdough fermentation processes, where the accumulation of acid by the growth of lactic acid bacteria reduces the yeast metabolic activity. In this work, we have carried out adaptive laboratory evolution (ALE) experiments in two sourdough isolates of S. cerevisiae exposed to acetic acid, or alternatively to acetic acid and myriocin, an inhibitor of sphingolipid biosynthesis that sped-up the evolutionary adaptation. Evolution approaches resulted in acetic tolerance, and surprisingly, increased lactic susceptibility. Four evolved clones, one from each parental strain and evolutionary scheme, were selected on the basis of their potential for CO2 production in sourdough conditions. Among them, two showed phenotypic instability characterized by strong lactic sensitivity after several rounds of growth under unstressed conditions, while two others, displayed increased constitutive acetic tolerance with no loss of growth in lactic medium. Genome sequencing and ploidy level analysis of all strains revealed aneuploidies, which could account for phenotypic heterogeneity. In addition, copy number variations (CNVs), affecting specially to genes involved in ion transport or flocculation, and single nucleotide polymorphisms (SNPs) were identified. Mutations in several genes, ARG82, KEX1, CTK1, SPT20, IRA2, ASG1 or GIS4, were confirmed as involved in acetic and/or lactic tolerance, and new determinants of these phenotypes, MSN5 and PSP2, identified.

Slt2 Is Required to Activate ER-Stress-Protective Mechanisms through TORC1 Inhibition and Hexosamine Pathway Activation.

Slt2, the MAPK of the cell wall integrity (CWI) pathway, connects different signaling pathways and performs different functions in the protective response of S. cerevisiae to stress. Previous work has evidenced the relation of the CWI pathway and the unfolded protein response (UPR), a transcriptional program activated upon endoplasmic reticulum (ER) stress. However, the mechanisms of crosstalk between these pathways and the targets regulated by Slt2 under ER stress remain unclear. Here, we demonstrated that ectopic expression of GFA1, the gene encoding the first enzyme in the synthesis of UDP-GlcNAc by the hexosamine biosynthetic pathway (HBP) or supplementation of the growth medium with glucosamine (GlcN), increases the tolerance of slt2 mutant cells to different ER-stress inducers. Remarkably, GlcN also alleviates the sensitivity phenotype of cells lacking IRE1 or HAC1, the main actors in controlling the UPR. The exogenous addition of GlcN reduced the abundance of glycosylated proteins and triggered autophagy. We also found that TORC1, the central stress and growth controller, is inhibited by tunicamycin exposure in cells of the wild-type strain but not in those lacking Slt2. Consistent with this, the tunicamycin-induced activation of autophagy and the increased synthesis of ATP in response to ER stress were absent by knock-out of SLT2. Altogether, our data placed Slt2 as an essential actor of the ER stress response by regulating the HBP activity and the TORC1-dependent signaling.

IGF-1 facilitates extinction of conditioned fear.

Insulin-like growth factor-1 (IGF-1) plays a key role in synaptic plasticity, spatial learning, and anxiety-like behavioral processes. While IGF-1 regulates neuronal firing and synaptic transmission in many areas of the central nervous system, its signaling and consequences on excitability, synaptic plasticity, and animal behavior dependent on the prefrontal cortex remain unexplored. Here, we show that IGF-1 induces a long-lasting depression of the medium and slow post-spike afterhyperpolarization (mAHP and sAHP), increasing the excitability of layer 5 pyramidal neurons of the rat infralimbic cortex. Besides, IGF-1 mediates a presynaptic long-term depression of both inhibitory and excitatory synaptic transmission in these neurons. The net effect of this IGF-1-mediated synaptic plasticity is a long-term potentiation of the postsynaptic potentials. Moreover, we demonstrate that IGF-1 favors the fear extinction memory. These results show novel functional consequences of IGF-1 signaling, revealing IGF-1 as a key element in the control of the fear extinction memory.

Sphingolipids and Inositol Phosphates Regulate the Tau Protein Phosphorylation Status in Humanized Yeast.

Hyperphosphorylation of protein tau is a hallmark of Alzheimer's disease (AD). Changes in energy and lipid metabolism have been correlated with the late onset of this neurological disorder. However, it is uncertain if metabolic dysregulation is a consequence of AD or one of the initiating factors of AD pathophysiology. Also, it is unclear whether variations in lipid metabolism regulate the phosphorylation state of tau. Here, we show that in humanized yeast, tau hyperphosphorylation is stimulated by glucose starvation in coincidence with the downregulation of Pho85, the yeast ortholog of CDK5. Changes in inositol phosphate (IP) signaling, which has a central role in energy metabolism, altered tau phosphorylation. Lack of inositol hexakisphosphate kinases Kcs1 and Vip1 (IP6 and IP7 kinases in mammals) increased tau hyperphosphorylation. Similar effects were found by mutation of IPK2 (inositol polyphosphate multikinase), or PLC1, the yeast phospholipase C gene. These effects may be explained by IP-mediated regulation of Pho85. Indeed, this appeared to be the case for plc1, ipk2, and kcs1. However, the effects of Vip1 on tau phosphorylation were independent of the presence of Pho85, suggesting additional mechanisms. Interestingly, kcs1 and vip1 strains, like pho85, displayed dysregulated sphingolipid (SL) metabolism. Moreover, genetic and pharmacological inhibition of SL biosynthesis stimulated the appearance of hyperphosphorylated forms of tau, while increased flux through the pathway reduced its abundance. Finally, we demonstrated that Sit4, the yeast ortholog of human PP2A protein phosphatase, is a downstream effector of SL signaling in mediating the tau phosphorylation state. Altogether, our results add new knowledge on the molecular effectors involved in tauopathies and identify new targets for pharmacological intervention.

Myriocin-induced adaptive laboratory evolution of an industrial strain of Saccharomyces cerevisiae reveals its potential to remodel lipid composition and heat tolerance.

The modification of lipid composition allows cells to adjust membrane biophysical properties in response to changes in environmental temperature. Here, we use adaptive laboratory evolution (ALE) in the presence of myriocin, a sphingolipid (SLs) biosynthesis inhibitor, to remodel the lipid profile of an industrial yeast strain (LH) of Saccharomyces cerevisiae. The approach enabled to obtain a heterogeneous population (LHev) of myriocin-tolerant evolved clones characterized by its growth capacity at high temperature. Myriocin exposure also caused tolerance to soraphen A, an inhibitor of the acetyl-CoA carboxylase Acc1, the rate-limiting enzyme in fatty acid de novo production, supporting a change in lipid metabolism during ALE. In line with this, characterization of two randomly selected clones, LH03 and LH09, showed the presence of lipids with increased saturation degree and reduced acyl length. In addition, the clone LH03, which displays the greater improvement in fitness at 40°C, exhibited higher SL content as compared with the parental strain. Analysis of the LH03 and LH09 genomes revealed a loss of chromosomes affecting genes that have a role in fatty acid synthesis and elongation. The link between ploidy level and growth at high temperature was further supported by the analysis of a fully isogenic set of yeast strains with ploidy between 1N and 4N which showed that the loss of genome content provides heat tolerance. Consistent with this, a thermotolerant evolved population (LH40°) generated from the parental LH strain by heat-driven ALE exhibited a reduction in the chromosome copy number. Thus, our results identify myriocin-driven evolution as a powerful approach to investigate the mechanisms of acquired thermotolerance and to generate improved strains.

Pho85 and PI(4,5)P2 regulate different lipid metabolic pathways in response to cold.

Lipid homeostasis allows cells to adjust membrane biophysical properties in response to changes in environmental conditions. In the yeast Saccharomyces cerevisiae, a downward shift in temperature from an optimal reduces membrane fluidity, which triggers a lipid remodeling of the plasma membrane. How changes in membrane fluidity are perceived, and how the abundance and composition of different lipid classes is properly balanced, remain largely unknown. Here, we show that the levels of phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2], the most abundant plasma membrane phosphoinositide, drop rapidly in response to a downward shift in temperature. This change triggers a signaling cascade transmitted to cytosolic diphosphoinositol phosphate derivatives, among them 5-PP-IP4 and 1-IP7, that exert regulatory functions on genes involved in the inositol and phospholipids (PLs) metabolism, and inhibit the activity of the protein kinase Pho85. Consistent with this, cold exposure triggers a specific program of neutral lipids and PLs changes. Furthermore, we identified Pho85 as playing a key role in controlling the synthesis of long-chain bases (LCBs) via the Ypk1-Orm2 regulatory circuit. We conclude that Pho85 orchestrates a coordinated response of lipid metabolic pathways that ensure yeast thermal adaptation.

The formation of hybrid complexes between isoenzymes of glyceraldehyde-3-phosphate dehydrogenase regulates its aggregation state, the glycolytic activity and sphingolipid status in Saccharomyces cerevisiae.

The glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has been traditionally considered a housekeeping protein involved in energy generation. However, evidence indicates that GAPDHs from different origins are tightly regulated and that this regulation may be on the basis of glycolysis-related and glycolysis-unrelated functions. In Saccharomyces cerevisiae, Tdh3 is the main GAPDH, although two other isoenzymes encoded by TDH1 and TDH2 have been identified. Like other GAPDHs, Tdh3 exists predominantly as a tetramer, although dimeric and monomeric forms have also been isolated. Mechanisms of Tdh3 regulation may thus imply changes in its oligomeric state or be based in its ability to interact with Tdh1 and/or Tdh2 to form hybrid complexes. However, no direct evidence of the existence of these interactions has been provided and the exact function of Tdh1,2 is unknown. Here, we show that Tdh1,2 immunopurified with a GFP-tagged version of Tdh3 and that lack of this interaction stimulates the Tdh3's aggregation. Furthermore, we found that the combined knockout of TDH1 and TDH2 promotes the loss of cell's viability and increases the growing rate, glucose consumption and CO2 production, suggesting a higher glycolytic flux in the mutant cells. Consistent with this, the tdh3 strain, which displays impaired in vitro GAPDH activity, exhibited the opposite phenotypes. Quite remarkably, tdh1 tdh2 mutant cells show increased sensitivity to aureobasidin A, an inhibitor of the inositolphosphoryl ceramide synthase, while cells lacking Tdh3 showed improved tolerance. The results are in agreement with a link between glycolysis and sphingolipid (SLs) metabolism. Engineering Tdh activity could be thus exploited to alter the SLs status with consequences in different aspects of yeast biotechnology.

Reiterative epoxide-based strategies for the synthesis of stereo-n-ads and application to polypropionate synthesis. A Personal Account.

The enantioselective synthesis of polypropionates continues to be an attractive realm for the synthetic chemists mostly due to the challenges presented by the number of consecutive stereogenic centers contained within the aliphatic chain. Over the years, our laboratory has developed an epoxide-based three-step reiterative methodology for the construction of these targets, with the ultimate goal that the approach could be extended to the synthesis of polypropionate-containing natural products. The key steps include the diastereoselective epoxidation of allylic and homoallylic alcohols, and the regioselective cleavage of 2-methyl-3,4-epoxy alcohols. The choice of the organometallic reagent, and the cis/trans geometry of the chiral epoxide can be used to control both the relative and absolute configuration of the resulting propionate unit, allowing our approach to be applied in the synthesis of advanced fragments. Additionally, the combination of our first- and second-generation methodologies permits the incorporation of different variations at the methyl moiety.

Design and validation of a questionnaire to assess cooperative learning in educational contexts / Diseño y validación de un cuestionario de medición del aprendizaje cooperativo en contextos educativos

The goal of the present study was to design and validate an instrument to assess the basic elements of cooperative learning, as well as a cooperation index. 11.202 primary education (grades 5, 6), secondary education and baccalaureate students (5.838 males, 5.364 females) from 68 different schools in 62 cities all over Spain agreed to participate. The age range was 11-18 years. The participating students had experienced several cooperative learning techniques during the last six months. The first version of the questionnaire was assessed by a group of experts. A pilot study with 60 students similar to the target sample was conducted on the second version of the instrument. The final version underwent several statistical tests. The Cooperative Learning Questionnaire included five subscales: Promotive Interaction, Positive Interdependence, Individual Accountability, Group Processing and Interpersonal skills. Factorial and confirmatory analysis showed that all reliability indices were acceptable, even under the most difficult conditions. The questionnaire showed adequate convergent, discriminant and concurrent validity. Results showed that it is an easy instrument to assess all the basic elements of cooperative learning in primary, secondary and baccalaureate students and obtain a global cooperation factor (AU)

El objetivo del estudio fue elaborar y validar un instrumento que pudiera evaluar los elementos fundamentales del aprendizaje cooperativo, así como proporcionar un factor de cooperación. Participaron 11.202 estudiantes de educación primaria (5º-6º curso), secundaria y bachillerato (5.838 varones, 5.364 mujeres) de 68 centros educativos en 62 ciudades españolas repartidas por toda su geografía. Las edades oscilaron entre los 11 y los 18 años. El único requisito para participar era haber experimentado varias técnicas de aprendizaje cooperativo en los últimos 6 meses. Tras elaborar una primera versión y ser sometida sucesivamente a un juicio de expertos y un estudio piloto se realizó un segundo estudio en el que se sometió la versión definitiva a diferentes pruebas estadísticas. El Cuestionario de Aprendizaje Cooperativo está formado por cinco sub-escalas: Interacción Promotora, Interdependencia Positiva, Responsabilidad Individual, Procesamiento Grupal y Habilidades Sociales. Los análisis factoriales confirmatorios mostraron que todos los índices de fiabilidad eran aceptables, incluso bajo las condiciones más exigentes. El cuestionario mostró una adecuada validez convergente, discriminante y concurrente. Se confirma como un instrumento sencillo para evaluar todos los elementos fundamentales del aprendizaje cooperativo en estudiantes de primaria, secundaria y bachillerato y proporcionar un factor de cooperación global (AU)

Crystal structure of (-)-(S)-4-[(2S,3S,4S,Z)-3-hydroxy-4-methyl-hept-5-en-2-yl]-1,3-dioxolan-2-one.

The title compound, C11H18O4, consists of an anti,anti,anti-stereo-tetrad with a 1,2-carbonate and an alkene motif. The mol-ecule displays a common zigzag conformation. The five-membered ring has a twisted conformation on the C-C bond. In the crystal, a strong inter-molecular hydrogen bond between the hy-droxy group and the carboxyl-ate moiety from an adjacent mol-ecule forms chains propagating along the b-axis direction. The absolute structure of the mol-ecule in the crystal was determined by resonant scattering [Flack parameter = 0.05 (6)].

How can lifeguards recover better? A cross-over study comparing resting, running, and foam rolling.

PURPOSE: The aim of this study is to compare the effectiveness of active recovery in form of running or foam rolling on clearing blood lactate compared to remain sitting after a water rescue. METHOD: A quasi experimental cross-over design was used to test the effectiveness of two active recovery methods: foam rolling (FR) and running (RR), compared with passive recovery (PR) on the blood lactate clearance after performing a water rescue. Twelve lifeguards from Marín (Pontevedra) completed the study. The participants performed a 100-meter water rescue and a 25-minute recovery protocol. RESULTS: The post recovery lactate levels were significantly lower for foam rolling (4.4±1.5mmol/l, P=0.005, d=0.94) and running (4.9±2.3mmol/l, P=0.027, d=1.21) compared with resting (7.2±2.5mmol/l); there was no significant difference between foam rolling and running (P=1.000). CONCLUSIONS: We found that surf lifesavers clear out blood lactate more efficient when performing an active recovery protocol. Foam rolling is an effective method of increasing the rate of blood lactate clearance. These two recovery methods are also adequate for surf lifeguards as they do not interfere with the surveillance aspect of their job.

Self-Regulation, Cooperative Learning, and Academic Self-Efficacy: Interactions to Prevent School Failure.

Learning to learn and learning to cooperate are two important goals for individuals. Moreover, self regulation has been identified as fundamental to prevent school failure. The goal of the present study was to assess the interactions between self-regulated learning, cooperative learning and academic self-efficacy in secondary education students experiencing cooperative learning as the main pedagogical approach for at least one school year. 2.513 secondary education students (1.308 males, 1.205 females), 12-17 years old (M = 13.85, SD = 1.29), enrolled in 17 different schools belonging to the National Network of Schools on Cooperative Learning in Spain agreed to participate. They all had experienced this pedagogical approach a minimum of one school year. Participants were asked to complete the cooperative learning questionnaire, the strategies to control the study questionnaire and the global academic self-efficacy questionnaire. Participants were grouped based on their perceptions on cooperative learning and self-regulated learning in their classes. A combination of hierarchical and κ-means cluster analyses was used. Results revealed a four-cluster solution: cluster one included students with low levels of cooperative learning, self-regulated learning and academic self-efficacy, cluster two included students with high levels of cooperative learning, self-regulated learning and academic self-efficacy, cluster three included students with high levels of cooperative learning, low levels of self-regulated learning and intermediate-low levels of academic self-efficacy, and, finally, cluster four included students with high levels of self-regulated learning, low levels of cooperative learning, and intermediate-high levels of academic self-efficacy. Self-regulated learning was found more influential than cooperative learning on students' academic self-efficacy. In cooperative learning contexts students interact through different types of regulations: self, co, and shared. Educators should be aware of these interactions, symmetrical or asymmetrical, because they determine the quality and quantity of the students' participation and achievements, and they are key elements to prevent school failure.

The Antarctic yeast Candida sake: Understanding cold metabolism impact on wine.

Current winemaking trends include low-temperature fermentations and using non-Saccharomyces yeasts as the most promising tools to produce lower alcohol and increased aromatic complexity wines. Here we explored the oenological attributes of a C. sake strain, H14Cs, isolated in the sub-Antarctic region. As expected, the cold sea water yeast strain showed greater cold growth, Na+-toxicity resistance and freeze tolerance than the S. cerevisiae QA23 strain, which we used as a commercial wine yeast control. C. sake H14Cs was found to be more sensitive to ethanol. The fermentation trials of low-sugar content must demonstrated that C. sake H14Cs allowed the cold-induced lag phase of growth to be eliminated and also notably reduced the ethanol (-30%) and glycerol (-50%) content in wine. Instead C. sake produced sorbitol as a compatible osmolyte. Finally, the inspection of the main wine volatile compounds revealed that C. sake produced more higher alcohols than S. cerevisiae. In conclusion, our work evidences that using the Antarctic C. sake H14Cs yeast improves low-temperature must fermentations and has the potential to provide a wine with less ethanol and also particular attributes.

Inappropriate translation inhibition and P-body formation cause cold-sensitivity in tryptophan-auxotroph yeast mutants.

In response to different adverse conditions, most eukaryotic organisms, including Saccharomyces cerevisiae, downregulate protein synthesis through the phosphorylation of eIF2α (eukaryotic initiation factor 2α) by Gcn2, a highly conserved protein kinase. Gcn2 also controls the translation of Gcn4, a transcription factor involved in the induction of amino acid biosynthesis enzymes. Here, we have studied the functional role of Gcn2 and Gcn2-regulating proteins, in controlling translation during temperature downshifts of TRP1 and trp1 yeast cells. Our results suggest that neither cold-instigated amino acid limitation nor Gcn2 are involved in the translation suppression at low temperature. However, loss of TRP1 causes increased eIF2α phosphorylation, Gcn2-dependent polysome disassembly and overactivity of Gcn4, which result in cold-sensitivity. Indeed, knock-out of GCN2 improves cold growth of trp1 cells. Likewise, mutation of several Gcn2-regulators and effectors results in cold-growth effects. Remarkably, we found that Hog1, the osmoresponsive MAPK, plays a role in the regulatory mechanism of Gcn2-eIF2α. Finally, we demonstrated that P-body formation responds to a downshift in temperature in a TRP1-dependent manner and is required for cold tolerance.

Sng1 associates with Nce102 to regulate the yeast Pkh-Ypk signalling module in response to sphingolipid status.

All cells are delimited by biological membranes, which are consequently a primary target of stress-induced damage. Cold alters membrane functionality by decreasing lipid fluidity and the activity of membrane proteins. In Saccharomyces cerevisiae, evidence links sphingolipid homeostasis and membrane phospholipid asymmetry to the activity of the Ypk1/2 proteins, the yeast orthologous of the mammalian SGK1-3 kinases. Their regulation is mediated by different protein kinases, including the PDK1 orthologous Pkh1/2p, and requires the function of protein effectors, among them Nce102p, a component of the sphingolipid sensor machinery. Nevertheless, the mechanisms and the actors involved in Pkh/Ypk regulation remain poorly defined. Here, we demonstrate that Sng1, a transmembrane protein, is an effector of the Pkh/Ypk module and identify the phospholipid asymmetry as key for yeast cold adaptation. Overexpression of SNG1 impairs phospholipid flipping, reduces reactive oxygen species (ROS) and improves, in a Pkh-dependent manner, yeast growth in myriocin-treated cells, suggesting that excess Sng1p stimulates the Pkh/Ypk signalling. Furthermore, we link these effects to the association of Sng1p with Nce102p. Indeed, we found that Sng1p interacts with Nce102p both physically and genetically. Moreover, mutant nce102∆ sng1∆ cells show features of impaired Pkh/Ypk signalling, including increased ROS accumulation, reduced life span and defects in Pkh/Ypk-controlled regulatory pathways. Finally, myriocin-induced hyperphosphorylation of Ypk1p and Orm2p, which controls sphingolipid homeostasis, does not occur in nce102∆ sng1∆ cells. Hence, both Nce102p and Sng1p participate in a regulatory circuit that controls the activity of the Pkh/Ypk module and their function is required in response to sphingolipid status.

Genista anglica (Fabaceae): One very diverse species or one species complex?

Genista anglica represents a widely distributed group of shrubs in the Iberian Peninsula, as well as in the North of the Moroccan Mountains, the South of Italy and in most oceanic territories of Western Europe, with its northern limit in Sweden. Up to five different species within the group have been described in these territories: Genista ancistrocarpa, G. acutifolia, G. brutia and G. silana, as well as G. anglica sensu stricto. The diversity of Genista anglica sensu lato as well as the phylogenetic patterns that have generated this diversity have been analyzed through the use of nuclear (ITS, ETS) and chloroplastic (trnL, trnL-F, rbcL, matK) DNA sequences. Our results show that the group probably originated in the West of the Iberian Peninsula and subsequently spread to the rest of the European oceanic territories. Additionally, the results support the idea that the presence of a group of plants in the South of Italy, where G. brutia and G. silana were previously described, has been the consequence of the introduction of seeds collected in the Northwest of the Iberian Peninsula. Our results also indicate that, in contrast to some authors, the populations from the West of the Iberian Peninsula are not isolated and, consequently, they should be grouped into one species with high diversity, therefore differentiation into different taxa is no longer adequate.