Genetic improvement of non-conventional Torulaspora delbrueckii for traditional sparkling winemaking by mixing for eventual hybridization with Saccharomyces cerevisiae.

Non-conventional yeasts such as Torulaspora delbrueckii (Td) have been proposed for sparkling winemaking. Unfortunately, this yeast has poor efficiency in completing wine fermentation as compared to Saccharomyces cerevisiae (Sc). New mutants with increased resistance to SO2, ethanol, and high CO2 pressure were previously isolated from spore clones of Td. Although these mutants showed improved capability for base wine fermentation, there is still room for genetic improvement of Td yeasts until the fermentative capacity of Sc is achieved. As an alternative approach, yeast mixture for eventual hybridization of Td with Sc was assayed in this study. The new yeast mixture clones (Sc-mixed Td) showed an intermediate phenotype between both parent yeasts for some relevant biotechnological properties, such as resistance to SO2, ethanol, copper, high CO2 pressure, and high temperature, as well as flocculation potential. These properties varied depending on the specific Sc-mixed Td clone. Several mixture clones showed improved capability for base wine fermentation as compared to the Td parent strain, approaching the fermentation capability of the Sc parent strain. The organoleptic quality of sparkling wine was also improved by using some mixture clones and this improved quality coincided with an increased amount of acetate and ethyl esters. The genetic stability of some Sc-mixed Td clones was good enough for commercial yeast production and winery applications.

Enhanced ammonia detoxification to urea in hepatocytes transduced with human aquaporin-8 gene.

Hepatic ammonia detoxification to urea is critical for the prevention of hyperammonemia and neurological damage. Hepatocyte mitochondrial aquaporin-8 (AQP8) channels have been involved in ammonia-derived ureagenesis. Herein, we studied whether the adenoviral gene transfer of human AQP8 (hAQP8) to hepatocyte mitochondria enhances ammonia conversion to urea. Using primary cultured rat hepatocytes, we first confirmed the mitochondrial expression of hAQP8 and then, using unlabeled or 15 N-labeled ammonia, we demonstrated that the urea synthesis was significantly enhanced in hAQP8-transduced hepatocytes. Studies using isolated hAQP8-expressing mitochondria also showed an increased ammonia metabolism. hAQP8 transduction was able to recover the impaired ammonia-derived ureagenesis in hepatotoxin-treated hepatocytes. Our data suggest that mitochondrially-expressed hAQP8 enhances and improves hepatocyte ammonia conversion to urea, a finding with potential therapeutic implications for liver disease with impaired ammonia detoxification.

Genetic Improvement of Torulaspora delbrueckii for Wine Fermentation: Eliminating Recessive Growth-Retarding Alleles and Obtaining New Mutants Resistant to SO2, Ethanol, and High CO2 Pressure.

The use of Torulaspora delbrueckii has been repeatedly proposed to improve a wine's organoleptic quality. This yeast has lower efficiency in completing wine fermentation than Saccharomyces cerevisiae since it has less fermentation capability and greater sensitivity to SO2, ethanol, and CO2 pressure. Therefore, the completion of fermentation is not guaranteed when must or wine is single-inoculated with T. delbrueckii. To solve this problem, new strains of T. delbrueckii with enhanced resistance to winemaking conditions were obtained. A genetic study of four wine T. delbrueckii strains was carried out. Spore clones free of possible recessive growth-retarding alleles were obtained from these yeasts. These spore clones were used to successively isolate mutants resistant to SO2, then those resistant to ethanol, and finally those resistant to high CO2 pressure. Most of these mutants showed better capability for base wine fermentation than the parental strain, and some of them approached the fermentation capability of S. cerevisiae. The genetic stability of the new mutants was good enough to be used in industrial-level production in commercial wineries. Moreover, their ability to ferment sparkling wine could be further improved by the continuous addition of oxygen in the culture adaptation stage prior to base wine inoculation.

Long-term Noninvasive Ventilation in Obesity Hypoventilation Syndrome Without Severe OSA: The Pickwick Randomized Controlled Trial.

BACKGROUND: Noninvasive ventilation (NIV) is an effective form of treatment in obesity hypoventilation syndrome (OHS) with severe OSA. However, there is paucity of evidence in patients with OHS without severe OSA phenotype. RESEARCH QUESTION: Is NIV effective in OHS without severe OSA phenotype? STUDY DESIGN AND METHODS: In this multicenter, open-label parallel group clinical trial performed at 16 sites in Spain, we randomly assigned 98 stable ambulatory patients with untreated OHS and apnea-hypopnea index < 30 events/h (ie, no severe OSA) to NIV or lifestyle modification (control group) using simple randomization through an electronic database. The primary end point was hospitalization days per year. Secondary end points included other hospital resource utilization, incident cardiovascular events, mortality, respiratory functional tests, BP, quality of life, sleepiness, and other clinical symptoms. Both investigators and patients were aware of the treatment allocation; however, treating physicians from the routine care team were not aware of patients' enrollment in the clinical trial. The study was stopped early in its eighth year because of difficulty identifying patients with OHS without severe OSA. The analysis was performed according to intention-to-treat and per-protocol principles and by adherence subgroups. RESULTS: Forty-nine patients in the NIV group and 49 in the control group were randomized, and 48 patients in each group were analyzed. During a median follow-up of 4.98 years (interquartile range, 2.98-6.62), the mean hospitalization days per year ± SD was 2.60 ± 5.31 in the control group and 2.71 ± 4.52 in the NIV group (adjusted rate ratio, 1.07; 95% CI, 0.44-2.59; P = .882). NIV therapy, in contrast with the control group, produced significant longitudinal improvement in Paco2, pH, bicarbonate, quality of life (Medical Outcome Survey Short Form 36 physical component), and daytime sleepiness. Moreover, per-protocol analysis showed a statistically significant difference for the time until the first ED visit favoring NIV. In the subgroup with high NIV adherence, the time until the first event of hospital admission, ED visit, and mortality was longer than in the low adherence subgroup. Adverse events were similar between arms. INTERPRETATION: In stable ambulatory patients with OHS without severe OSA, NIV and lifestyle modification had similar long-term hospitalization days per year. A more intensive program aimed at improving NIV adherence may lead to better outcomes. Larger studies are necessary to better determine the long-term benefit of NIV in this subgroup of OHS. TRIAL REGISTRY: ClinicalTrials.gov; No.: NCT01405976; URL: www.clinicaltrials.gov.

Using Torulaspora delbrueckii killer yeasts in the elaboration of base wine and traditional sparkling wine.

For still wines, killer strains of Torulaspora delbrueckii can be used instead of non-killer strains to improve this species' domination during must fermentation, with an ensured, reliable impact on the final wine quality. The present work analysed the usefulness of these killer yeasts for sparkling-wine making. After the first fermentation, the foaming capacity of T. delbrueckii base wines was very low compared to Saccharomyces cerevisiae base wines. Significant positive correlations of foaming parameters were found with the amounts of C4-C16 ethyl esters and proteins, and negative with some anti-foaming alcohols produced by each yeast species. There were, however, no evident positive effects of polysaccharides on those parameters. The organoleptic quality of the T. delbrueckii base wines was judged inappropriate for sparkling-wine making, so that the following second-fermentation experiments only used a single assemblage of S. cerevisiae base-wines. While second fermentation was completed with inoculation of S. cerevisiae (both alone and mixed with T. delbrueckii) to yield dry sparkling wines with high CO2 pressure, single inoculation with T. delbrueckii did not complete this fermentation, leaving sweet wines with poor CO2 pressure. Yeast death due to CO2 pressure was much greater in T. delbrueckii than in S. cerevisiae, making any killer effect of S. cerevisiae over T. delbrueckii irrelevant because no autolysed cells were found during the first days of mixed-inoculated second fermentation. Nonetheless, the organoleptic quality of the mixed-inoculated sparkling wines was better than that of wines single-inoculated with S. cerevisiae, and showed no deterioration in foam quality. This seemed mainly to be because T. delbrueckii increased the amounts of ethyl propanoate and some acids (e.g., isobutyric and butanoic), alcohols (e.g., 3­ethoxy­1­propanol), and phenols (e.g., 4­vinylguaiacol). For these sparkling wines, no significant correlations between foaming parameters and aroma compounds were found, probably because the differences in foaming parameter values among these wines were fairly small. This is unlike the case for the base wines for which there were large differences in these parameters, which facilitated the analysis of the influence of aroma compounds on base-wine foamability.

NSAIDs hypersensitivity: questions not resolved.

PURPOSE OF REVIEW: NSAIDs are the drugs most frequently involved in hypersensitivity reactions (HSR). These are frequently prescribed at all ages. HSR are of great concern and can affect people at any age. These drugs can induce reactions by stimulating the adaptive immune system (IgE or T cell), known as selective responders or more frequently by abnormalities in biochemical pathways related with prostaglandin metabolism. These are known as cross-intolerant. With some exceptions, skin testing and in-vitro studies are of little value in selective responders. RECENT FINDINGS: In the last years, several classifications have been provided based on clinical symptoms, time interval between drug intake and appearance of symptoms, response to other nonchemically related NSAIDs and the underlying disease. Based on this classification, several well differentiated categories within each group of entities cross-intolerant and selective responders are now recognized. The most complex groups for evaluation are cross-intolerant in which three major groups exist: NSAIDs exacerbated respiratory disease, NSAIDs exacerbated cutaneous disease and NSAIDs-induced urticaria/angioedema in the absence of chronic spontaneous urticaria. Within the selective responders, there are two mechanisms involved: drug-specific IgE or T-cell effector responses. New entities have been added to this classification like mixed reactions within the cross-intolerant category, that must manifest as anaphylaxis and multiple immediate selective reactions. SUMMARY: The precise evaluation of patients with NSAIDs hypersensitivity following established guidelines will improve not only our understanding but also the management of these entities. As the number of patients affected with NSAIDs is important, further studies are warranted.

Echocardiographic changes with non-invasive ventilation and CPAP in obesity hypoventilation syndrome.

RATIONALE: Despite a significant association between obesity hypoventilation syndrome (OHS) and cardiac dysfunction, no randomised trials have assessed the impact of non-invasive ventilation (NIV) or CPAP on cardiac structure and function assessed by echocardiography. OBJECTIVES: We performed a secondary analysis of the data from the largest multicentre randomised controlled trial of OHS (Pickwick project, n=221) to determine the comparative efficacy of 2 months of NIV (n=71), CPAP (n=80) and lifestyle modification (control group, n=70) on structural and functional echocardiographic changes. METHODS: Conventional transthoracic two-dimensional and Doppler echocardiograms were obtained at baseline and after 2 months. Echocardiographers at each site were blinded to the treatment arms. Statistical analysis was performed using intention-to-treat analysis. RESULTS: At baseline, 55% of patients had pulmonary hypertension and 51% had evidence of left ventricular hypertrophy. Treatment with NIV, but not CPAP, lowered systolic pulmonary artery pressure (-3.4 mm Hg, 95% CI -5.3 to -1.5; adjusted P=0.025 vs control and P=0.033 vs CPAP). The degree of improvement in systolic pulmonary artery pressure was greater in patients treated with NIV who had pulmonary hypertension at baseline (-6.4 mm Hg, 95% CI -9 to -3.8). Only NIV therapy decreased left ventricular hypertrophy with a significant reduction in left ventricular mass index (-5.7 g/m2; 95% CI -11.0 to -4.4). After adjusted analysis, NIV was superior to control group in improving left ventricular mass index (P=0.015). Only treatment with NIV led to a significant improvement in 6 min walk distance (32 m; 95% CI 19 to 46). CONCLUSION: In patients with OHS, medium-term treatment with NIV is more effective than CPAP and lifestyle modification in improving pulmonary hypertension, left ventricular hypertrophy and functional outcomes. Long-term studies are needed to confirm these results. TRIAL REGISTRATION NUMBER: Pre-results, NCT01405976 (https://clinicaltrials.gov/).

Automated Screening of Children With Obstructive Sleep Apnea Using Nocturnal Oximetry: An Alternative to Respiratory Polygraphy in Unattended Settings.

STUDY OBJECTIVES: Nocturnal oximetry has become known as a simple, readily available, and potentially useful diagnostic tool of childhood obstructive sleep apnea (OSA). However, at-home respiratory polygraphy (HRP) remains the preferred alternative to polysomnography (PSG) in unattended settings. The aim of this study was twofold: (1) to design and assess a novel methodology for pediatric OSA screening based on automated analysis of at-home oxyhemoglobin saturation (SpO2), and (2) to compare its diagnostic performance with HRP. METHODS: SpO2 recordings were parameterized by means of time, frequency, and conventional oximetric measures. Logistic regression models were optimized using genetic algorithms (GAs) for three cutoffs for OSA: 1, 3, and 5 events/h. The diagnostic performance of logistic regression models, manual obstructive apnea-hypopnea index (OAHI) from HRP, and the conventional oxygen desaturation index ≥ 3% (ODI3) were assessed. RESULTS: For a cutoff of 1 event/h, the optimal logistic regression model significantly outperformed both conventional HRP-derived ODI3 and OAHI: 85.5% accuracy (HRP 74.6%; ODI3 65.9%) and 0.97 area under the receiver operating characteristics curve (AUC) (HRP 0.78; ODI3 0.75) were reached. For a cutoff of 3 events/h, the logistic regression model achieved 83.4% accuracy (HRP 85.0%; ODI3 74.5%) and 0.96 AUC (HRP 0.93; ODI3 0.85) whereas using a cutoff of 5 events/h, oximetry reached 82.8% accuracy (HRP 85.1%; ODI3 76.7) and 0.97 AUC (HRP 0.95; ODI3 0.84). CONCLUSIONS: Automated analysis of at-home SpO2 recordings provide accurate detection of children with high pretest probability of OSA. Thus, unsupervised nocturnal oximetry may enable a simple and effective alternative to HRP and PSG in unattended settings.

The Effect of Supplemental Oxygen in Obesity Hypoventilation Syndrome.

STUDY OBJECTIVES: Low flow supplemental oxygen is commonly prescribed to patients with obesity hypoventilation syndrome (OHS). However, there is a paucity of data regarding its efficacy and safety. The objective of this study was to assess the medium-term treatment efficacy of adding supplemental oxygen therapy to commonly prescribed treatment modalities in OHS. METHODS: In this post hoc analysis of a previous randomized controlled trial, we studied 302 sequentially screened OHS patients who were randomly assigned to noninvasive ventilation, continuous positive airway pressure, or lifestyle modification. Outcomes at 2 mo included arterial blood gases, symptoms, quality of life, blood pressure, polysomnography, spirometry, 6-min walk distance, and hospital resource utilization. Statistical analysis comparing patients with and without oxygen therapy in the three treatment groups was performed using an intention-to-treat analysis. RESULTS: In the noninvasive ventilation group, supplemental oxygen reduced systolic blood pressure although this could be also explained by a reduction in body weight experienced in this group. In the continuous positive airway pressure group, supplemental oxygen increased the frequency of morning confusion. In the lifestyle modification group, supplemental oxygen increased compensatory metabolic alkalosis and decreased the apnea-hypopnea index during sleep. Oxygen therapy was not associated with an increase in hospital resource utilization in any of the groups. CONCLUSIONS: After 2 mo of follow-up, chronic oxygen therapy produced marginal changes that were insufficient to consider it, globally, as beneficial or deleterious. Because supplemental oxygen therapy did not increase hospital resource utilization, we recommend prescribing oxygen therapy to patients with OHS who meet criteria with close monitoring. Long-term studies examining outcomes such as incident cardiovascular morbidity and mortality are necessary. CLINICAL TRIALS REGISTRATION: Clinicaltrial.gov, ID: NCT01405976.

Non-invasive ventilation in obesity hypoventilation syndrome without severe obstructive sleep apnoea.

BACKGROUND: Non-invasive ventilation (NIV) is an effective form of treatment in patients with obesity hypoventilation syndrome (OHS) who have concomitant severe obstructive sleep apnoea (OSA). However, there is a paucity of evidence on the efficacy of NIV in patients with OHS without severe OSA. We performed a multicentre randomised clinical trial to determine the comparative efficacy of NIV versus lifestyle modification (control group) using daytime arterial carbon dioxide tension (PaCO2) as the main outcome measure. METHODS: Between May 2009 and December 2014 we sequentially screened patients with OHS without severe OSA. Participants were randomised to NIV versus lifestyle modification and were followed for 2 months. Arterial blood gas parameters, clinical symptoms, health-related quality of life assessments, polysomnography, spirometry, 6-min walk distance test, blood pressure measurements and healthcare resource utilisation were evaluated. Statistical analysis was performed using intention-to-treat analysis. RESULTS: A total of 365 patients were screened of whom 58 were excluded. Severe OSA was present in 221 and the remaining 86 patients without severe OSA were randomised. NIV led to a significantly larger improvement in PaCO2 of -6 (95% CI -7.7 to -4.2) mm Hg versus -2.8 (95% CI -4.3 to -1.3) mm Hg, (p<0.001) and serum bicarbonate of -3.4 (95% CI -4.5 to -2.3) versus -1 (95% CI -1.7 to -0.2 95% CI)  mmol/L (p<0.001). PaCO2 change adjusted for NIV compliance did not further improve the inter-group statistical significance. Sleepiness, some health-related quality of life assessments and polysomnographic parameters improved significantly more with NIV than with lifestyle modification. Additionally, there was a tendency towards lower healthcare resource utilisation in the NIV group. CONCLUSIONS: NIV is more effective than lifestyle modification in improving daytime PaCO2, sleepiness and polysomnographic parameters. Long-term prospective studies are necessary to determine whether NIV reduces healthcare resource utilisation, cardiovascular events and mortality. TRIAL REGISTRATION NUMBER: NCT01405976; results.

Using mixed inocula of Saccharomyces cerevisiae killer strains to improve the quality of traditional sparkling-wine.

The quality of traditional sparkling-wine depends on the aging process in the presence of dead yeast cells. These cells undergo a slow autolysis process thereby releasing some compounds, mostly colloidal polymers such as polysaccharides and mannoproteins, which influence the wine's foam properties and mouthfeel. Saccharomyces cerevisiae killer yeasts were tested to increase cell death and autolysis during mixed-yeast-inoculated second fermentation and aging. These yeasts killed sensitive strains in killer plate assays done under conditions of low pH and temperature similar to those used in sparkling-wine making, although some strains showed a different killer behaviour during the second fermentation. The fast killer effect improved the foam quality and mouthfeel of the mixed-inoculated wines, while the slow killer effect gave small improvements over single-inoculated wines. The effect was faster under high-pressure than under low-pressure conditions. Wine quality improvement did not correlate with the polysaccharide, protein, mannan, or aromatic compound concentrations, suggesting that the mouthfeel and foaming quality of sparkling wine are very complex properties influenced by other wine compounds and their interactions, as well as probably by the specific chemical composition of a given wine.

Conocimientos de los pediatras sobre la laringomalacia: ¿siempre es un proceso banal? / Pediatrician knowledge about laryngomalacia: is it always a banal process?

Introducción y objetivo: se diseñó una encuesta nacional para analizar el grado de conocimientos teóricos y la actitud de los pediatras ante la laringomalacia por la percepción de que algunos casos graves son infravalorados. Material y método: estudio transversal, descriptivo y analítico, de ámbito nacional, mediante encuesta online. Incluyó pediatras del área hospitalaria y Atención Primaria, diseñándose un cuestionario de 16 preguntas. Las variables recogidas fueron filiación, conocimientos generales, diagnóstico y manejo de la laringomalacia. Resultados: se contestaron 233 encuestas. La actitud mayoritaria ante un caso de laringomalacia era expectante (54,1%), no existiendo diferencias en función de la experiencia de los pediatras, ni por años trabajados ni por número de casos de laringomalacia atendidos. El grado de conocimientos generales de los encuestados fue superior al 89%, disminuyendo al 57% en conocimientos referidos a las laringomalacias graves. El 67% conocía las pruebas complementarias a aplicar en los casos graves y el 73,3% las posibles comorbilidades. El 85,6% coinciden que la ventilación no invasiva puede ser útil en pacientes con laringomalacia grave. Conclusiones: la encuesta muestra que la laringomalacia es una patología conocida, aunque existe disparidad en la actitud inicial a seguir y en las herramientas que utilizan los pediatras para basar su manejo. A pesar de que el grado de conocimiento general conceptual y diagnóstico es adecuado, este disminuye en conceptos más específicos que engloban a los casos de laringomalacia grave. Sería deseable la elaboración de guías clínicas y protocolos para estandarizar el manejo de la laringomalacia (AU)

Background and aims: a national survey was designed to analyze the knowledge and approach of pediatricians to laryngomalacia, due to the perception that some severe cases were overlooked. Materials and methods: this is a national, transversal, descriptive and analytical study, conducted through an on-line survey. We designed a 16 item questionnaire, and targeted both primary care and specialty pediatricians. The variables included professional data filiation, general knowledge, diagnosis and management of laryngomalacia. Results: 233 surveys were answered. The most common approach in a case of suspected laryngomalacia was expectant (54.1%), no differences were found in terms of pediatricians experience, neither by years of time worked, nor by the number of laryngomalacia cases previously assisted. Level of general knowledge shown by participants was higher than 89%, while it decreased to 57% when analyzing items covering severe laryngomalacia. 67% were aware of the additional tests to perform in severe cases, and 73.3% were alert for possible comorbidities. 85.6% agreed that non-invasive ventilation is useful in patients with severe laryngomalacia. Conclusions: the survey shows that laryngomalacia is a well-known condition, although there is not a clear consensus in the initial approach nor in the tools used by pediatricians to base its management. Though the knowledge in general concepts and diagnostics is suitable, it decreases in more specific concepts related to cases of severe laryngomalacia. It would be desirable that clinical guidelines and protocols were developed, to standardize management of laryngomalacia (AU)

Protective Cardiovascular Effect of Sleep Apnea Severity in Obesity Hypoventilation Syndrome.

BACKGROUND: Obesity hypoventilation syndrome (OHS) is associated with a high burden of cardiovascular morbidity (CVM) and mortality. The majority of patients with OHS have concomitant OSA, but there is a paucity of data on the association between CVM and OSA severity in patients with OHS. The objective of our study was to assess the association between CVM and OSA severity in a large cohort of patients with OHS. METHODS: In a cross-sectional analysis, we examined the association between OSA severity based on tertiles of oxygen desaturation index (ODI) and CVM in 302 patients with OHS. Logistic regression models were constructed to quantify the independent association between OSA severity and prevalent CVM after adjusting for various important confounders. RESULTS: The prevalence of CVM decreased significantly with increasing severity of OSA based on ODI as a continuous variable or ODI tertiles. This inverse relationship between OSA severity and prevalence of CVM was seen in the highest ODI tertile and it persisted despite adjustment for multiple confounders. Chronic heart failure had the strongest negative association with the highest ODI tertile. No significant CVM risk change was observed between the first and second ODI tertiles. Patients in the highest ODI tertile were younger, predominantly male, more obese, more hypersomnolent, had worse nocturnal and daytime gas exchange, lower prevalence of hypertension, better exercise tolerance, and fewer days hospitalized than patients in the lowest ODI tertile. CONCLUSIONS: In patients with OHS, the highest OSA severity phenotype was associated with reduced risk of CVM. This finding should guide the design of future clinical trials assessing the impact of interventions aimed at decreasing cardiovascular morbidity and mortality in patients with OHS. TRIAL REGISTRY: Clinicaltrial.gov; No.: NCT01405976; URL: www.clinicaltrials.gov.

Effects of new Torulaspora delbrueckii killer yeasts on the must fermentation kinetics and aroma compounds of white table wine.

Torulaspora delbrueckii is becoming widely recommended for improving some specific characteristics of wines. However, its impact on wine quality is still far from satisfactory at the winery level, mostly because it is easily replaced by Saccharomyces cerevisiae-like yeasts during must fermentation. New T. delbrueckii killer strains were here isolated and selected for winemaking. They killed S. cerevisiae yeasts and were able to dominate and complete the fermentation of sterile grape must. Sequential yeast inoculation of non-sterile white must with T. delbrueckii followed by S. cerevisiae did not ensure T. delbrueckii dominance or wine quality improvement. Only a single initial must inoculation at high cell concentrations allowed the T. delbrueckii killer strains to dominate and complete the must fermentation to reach above 11% ethanol, but not the non-killer strains. None of the wines underwent malolactic fermentation as long as the must had low turbidity and pH. Although no statistically significant differences were found in the wine quality score, the S. cerevisiae-dominated wines were preferred over the T. delbrueckii-dominated ones because the former had high-intensity fresh fruit aromas while the latter had lower intensity, but nevertheless nice and unusual dried fruit/pastry aromas. Except for ethyl propanoate and 3-ethoxy-1-propanol, which were more abundant in the T. delbrueckii-dominated wines, most of the compounds with fresh fruit odor descriptors, including those with the greatest odor activity values (isoamyl acetate, ethyl hexanoate, and ethyl octanoate), were more abundant in the S. cerevisiae-dominated wines. The low relative concentrations of these fruity compounds made it possible to detect in the T. delbrueckii-dominated wines the low-relative-concentration compounds with dried fruit and pastry odors. An example was γ-ethoxy-butyrolactone which was significantly more abundant in these wines than in those dominated by S. cerevisiae.

FoxO3a modulation and promotion of apoptosis by interferon-α2b in rat preneoplastic liver.

BACKGROUND: FoxO3a, a member of the FOXO family of transcription factors, is expressed in adult liver and modulates the expression of genes involved in apoptosis. FoxO3a is post-translationally regulated, negatively by PI3K/Akt and MAPK/Erk and positively by oxidative stress/JNK pathways. In previous works, we have demonstrated that interferon-α2b (IFN-α2b) induces apoptosis of hepatic preneoplastic foci through the production of reactive oxygen species (ROS). AIMS: To investigate the post-translational signal events triggered by the oxidative stress induced by IFN-α2b and the modulation of FoxO3a transcriptional activity during these events in rat preneoplastic liver. METHODS: Adult male Wistar rats were subjected to a two-phase model of hepatocarcinogenesis. A group of animals received IFN-α2b and another group received IFN-α2b and ascorbic acid (ASC), by intraperitoneal injection. Lipid peroxidation, immunohistochemistry, immunoblotting, co-immunoprecipitation and sqRT-PCR assays were performed to explore the role of ROS, JNK, Akt, Erk, FoxO3a, ß-catenin and PUMA in the IFN-α2b-mediated apoptotic mechanism. RESULTS: In vivo IFN-α2b treatment induced endogenous production of ROS which activated JNK. IFN-α2b blocked the activation of Akt and Erk, avoiding FoxO3a activity repression. Activated JNK was responsible for the nuclear translocation and transcriptional activity of FoxO3a which positively modulated the expression of PUMA, a proapoptotic player. In addition, nuclear FoxO3a competed for the nuclear ß-catenin associated to TCF, inhibiting the canonical Wnt signalling pathway. CONCLUSIONS: The data presented here propose a model in which in vivo IFN-α2b treatment induces nuclear translocation and transcriptional activity of FoxO3a, triggering the mitochondrial apoptotic pathway in hepatic preneoplastic foci.

Characterization, ecological distribution, and population dynamics of Saccharomyces sensu stricto killer yeasts in the spontaneous grape must fermentations of southwestern Spain.

Killer yeasts secrete protein toxins that are lethal to sensitive strains of the same or related yeast species. Among the four types of Saccharomyces killer yeasts already described (K1, K2, K28, and Klus), we found K2 and Klus killer yeasts in spontaneous wine fermentations from southwestern Spain. Both phenotypes were encoded by medium-size double-stranded RNA (dsRNA) viruses, Saccharomyces cerevisiae virus (ScV)-M2 and ScV-Mlus, whose genome sizes ranged from 1.3 to 1.75 kb and from 2.1 to 2.3 kb, respectively. The K2 yeasts were found in all the wine-producing subareas for all the vintages analyzed, while the Klus yeasts were found in the warmer subareas and mostly in the warmer ripening/harvest seasons. The middle-size isotypes of the M2 dsRNA were the most frequent among K2 yeasts, probably because they encoded the most intense K2 killer phenotype. However, the smallest isotype of the Mlus dsRNA was the most frequent for Klus yeasts, although it encoded the least intense Klus killer phenotype. The killer yeasts were present in most (59.5%) spontaneous fermentations. Most were K2, with Klus being the minority. The proportion of killer yeasts increased during fermentation, while the proportion of sensitive yeasts decreased. The fermentation speed, malic acid, and wine organoleptic quality decreased in those fermentations where the killer yeasts replaced at least 15% of a dominant population of sensitive yeasts, while volatile acidity and lactic acid increased, and the amount of bacteria in the tumultuous and the end fermentation stages also increased in an unusual way.

Propuesta metodológica para desarrollar correctas prácticas bioéticas en estudiantes de Estomatología / Methodological proposal to develop correct bioethical practices in the Stomatology students

El proceso de enseñanza-aprendizaje debe concebirse como un espacio donde los valores, la moral y la ética propios de la profesión se conjugan con los aspectos técnicos académicos. Se realiza un estudio descriptivo de corte transversal sobre el nivel de conocimientos de los estudiantes acerca de los principios bioéticos durante la atención al paciente con VIH/SIDA, y se diseña una propuesta de orientaciones metodológicas para la adquisición de correctas prácticas bioéticas durante su atención ambulatoria. El universo del estudio estuvo conformado por los 203 estudiantes de 3ro, 4to y 5to años de la Facultad de Estomatología del curso 2009-2010. La muestra fue de tipo probabilística estratificada aleatoria constituida por 103 estudiantes. Los resultados demostraron que el nivel de conocimientos con relación a la aplicación de los principios bioéticos fue bajo y se elabora una propuesta que intenciona la formación de correctas prácticas bioéticas en los egresados de Estomatología(AU)

A low-cost procedure for production of fresh autochthonous wine yeast.

A low-cost procedure was designed for easy and rapid response-on-demand production of fresh wine yeast for local wine-making. The pilot plant produced fresh yeast culture concentrate with good microbial quality and excellent oenological properties from four selected wine yeasts. The best production yields were obtained using 2% sugar beet molasses and a working culture volume of less than 60% of the fermenter capacity. The yeast yield using 2% sugar grape juice was low and had poor cell viability after freeze storage, although the resulting yeast would be directly available for use in the winery. The performance of these yeasts in commercial wineries was excellent; they dominated must fermentation and improved its kinetics, as well as improving the physicochemical parameters and the organoleptic quality of red and white wines.

Attenuation of the Wnt/beta-catenin/TCF pathway by in vivo interferon-alpha2b (IFN-alpha2b) treatment in preneoplastic rat livers.

Wnt/beta-catenin/T cell factor (TCF) pathway is activated in several types of human cancers, promoting cell growth and proliferation. Forkhead box containing protein class O (FOXO) transcription factors compete with TCF for beta-catenin binding, particularly under cellular oxidative stress conditions. Contrary to beta-catenin/TCF, beta-catenin/FOXO promotes the transcription of genes involved in cell cycle arrest and apoptosis. We have previously demonstrated that in vivo interferon-alpha2b (IFN-alpha2b) administration induces apoptosis in preneoplastic livers, a mechanism mediated by reactive oxygen species (ROS) and transforming growth factor-beta(1) (TGF-beta(1)). This study was aimed to assess the status of the Wnt/beta-catenin/TCF pathway in a very early stage of rat hepatocarcinogenesis and to further evaluate the effects of in vivo IFN-alpha2b treatment on it. We demonstrated that the Wnt/beta-catenin/TCF pathway is activated in preneoplastic rat livers. More important, in vivo IFN-alpha2b treatment inhibits Wnt/beta-catenin/TCF pathway and promotes programed cell death possibly providing a link with FOXO pathway.

Tumor necrosis factor alpha induced by Trypanosoma cruzi infection mediates inflammation and cell death in the liver of infected mice.

Trypanosoma cruzi (T. cruzi) infected C57BL/6 mice developed a progressive fatal disease due to an imbalance in the profile of circulating related compounds accompanying infection like tumor necrosis factor alpha (TNFalpha). TNFalpha has been proposed as an important effector molecule in apoptosis. In this work, we evaluate inflammation and the proteins involved in apoptotic process in liver of infected mice and the role of TNFalpha. C57BL6/mice were infected subcutaneously with 100 viable trypomastigotes of Tulahuén strain of T cruzi. One set of these animals were treated with 375 microg of antihuman TNFalpha blocking antibody. Animals were sacrificed at 14 days post-infection (p.i).The analyses of Bcl-2 family proteins revealed an increase of the pro-apoptotic proteins Bax and tBid in T. cruzi-infected mice. Compared with control animals, cytochrome c release was increased. Apoptosis was also induced in infected mice. Anti-TNFalpha treatment decreases hepatic apoptosis. Our results suggest that T. cruzi infection induces programmed cell death in the host liver by increase of TNFalpha production, associated with TNF-R1 over-expression, that set in motion the Bid cleavage and mitochondrial translocation, Bax mitochondrial translocation, cytochrome c release, and ultimately apoptosis induction.