Controlled mixed fermentation at winery scale using Zygotorulaspora florentina and Saccharomyces cerevisiae

Over the last few years the use of multi-starter inocula has become an attractive biotechnological practice in the search for winewith high flavour complexity or distinctive characters. This has been possible through exploiting the particular oenological features of somenon-Saccharomyces yeast strains, and the effects that derive fromtheir specific interactions with Saccharomyces. In the present study, we evaluated the selected strain Zygotorulaspora florentina (formerly Zygosaccharomyces florentinus) in mixed culture fermentations with Saccharomyces cerevisiae, from the laboratory scale to the winery scale. The scale-up fermentation and substrate composition (i.e., white or red musts) influenced the analytical composition of the mixed fermentation. At the laboratory scale, mixed fermentation with Z. florentina exhibited an enhancement of polysaccharides and 2-phenylethanol content and a reduction of volatile acidity. At the winery scale, different fermentation characteristics of Z. florentina were observed. Using Sangiovese red grape juice, sequential fermentation trials showed a significantly higher concentration of glycerol and esters while the sensorial analysis of the resulting wines showed higher floral notes and lower perception of astringency. To our knowledge, this is the first time that this yeasts association has been evaluated at the winery scale indicating the potential use of this mixed culture in red grape varietie

Potential spoilage non- Saccharomyces yeasts in mixed cultures with Saccharomyces cerevisiae

With the aim of exploring the possibility to improve wine quality through the utilization of wine-related yeasts generally considered as spoilage, mixed cultures of Saccharomyces cerevisiae with Hanseniaspora osmophila, Pichia fermentans, Saccharomycodes ludwigii and Zygosaccharomyces bailii were inoculated in grape juice. All the fermentations got to completion and most of the compounds normally produced at high concentrations by pure cultures of non-Saccharomyces yeasts, and considered detrimental for wine quality, did not reach the threshold taste level in mixed fermentations with S. cerevisiae. Interestingly, the association of S. cerevisiae with P. fermentans, S. ludwigii and Z. bailii produced significant increases in the production of polysaccharides as compared to pure cultures of S. cerevisiae. Since polysaccharides improve wine taste and body, and exert positive effects on aroma persistence and protein and tartrate stability, a possible use for these yeasts can be envisaged in mixed starter cultures with S. cerevisiae for the enhancement of the final quality of wine

Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation

Treatment-free remission (TFR) by tyrosine kinase inhibitors (TKI) discontinuation in patients with deep molecular response (DMR) is a paramount goal in the current chronic myeloid leukemia (CML) therapeutic strategy. The best DMR level by real-time quantitative PCR (RT-qPCR) for TKI discontinuation is still a matter of debate. To compare the accuracy of digital PCR (dPCR) and RT-qPCR for BCR-ABL1 transcript levels detection, 142 CML patients were monitored for a median time of 24\ua0months. Digital PCR detected BCR-ABL1 transcripts in the RT-qPCR undetectable cases. The dPCR analysis of the samples, grouped by the MR classes, revealed a significant difference between MR4.0 and MR4.5 (P\ua0=\ua00.0104) or MR5.0 (P\ua0=\ua00.0032). The clinical and hematological characteristics of the patients grouped according to DMR classes (MR4.0 vs MR4.5-5.0 ) were superimposable. Conversely, patients with dPCR values <0.468 BCR-ABL1 copies/\ub5L (as we previously described) showed a longer DMR duration (P\ua0=\ua00.0220) and mainly belonged to MR4.5-5.0 (P\ua0=\ua00.0442) classes compared to patients with higher dPCR values. Among the 142 patients, 111 (78%) discontinued the TKI treatment; among the 111 patients, 24 (22%) lost the MR3.0 or MR4.0 . RT-qPCR was not able to discriminate patients with higher risk of MR loss after discontinuation (P\ua0=\ua00.8100). On the contrary, according to dPCR, 12/25 (48%) patients with BCR-ABL1 values 650.468 and 12/86 (14%) patients with BCR-ABL1 values <0.468 lost DMR in this cohort, respectively (P\ua0=\ua00.0003). Treatment-free remission of patients who discontinued TKI with a dPCR <0.468 was significantly higher compared to patients with dPCR\ua0 65\ua00.468 (TFR at 2\ua0years 83% vs 52% P\ua0=\ua00.0017, respectively). In conclusion, dPCR resulted in an improved recognition of stable DMR and of candidates to TKI discontinuation

Melatonin MT1 receptors as a target for the psychopharmacology of bipolar disorder: a translational study

The treatment of bipolar disorder (BD) still remains a challenge. Melatonin (MLT), acting through its two receptors MT1 and MT2, plays a key role in regulating circadian rhythms which are dysfunctional in BD. Using a translational approach, we examined the implication and potential of MT1 receptors in the pathophysiology and psychopharmacology of BD. We employed a murine model of the manic phase of BD (Clock mutant (ClockΔ19) mice) to study the activation of MT1 receptors by UCM871, a selective partial agonist, in behavioral pharmacology tests and in-vivo electrophysiology. We then performed a high-resolution Nuclear Magnetic Resonance study on isolated membranes to characterize the molecular mechanism of interaction of UCM871. Finally, in a cohort of BD patients, we investigated the link between clinical measures of BD and genetic variants located in the MT1 receptor and CLOCK genes. We demonstrated that: 1) UCM871 can revert behavioral and electrophysiological abnormalities of ClockΔ19 mice; 2) UCM871 promotes the activation state of MT1 receptors; 3) there is a significant association between the number of severe manic episodes and MLT levels, depending on the genetic configuration of the MT1 rs2165666 variant. Overall, this work lends support to the potentiality of MT1 receptors as target for the treatment of BD

Synergistic Interactions between HDAC and Sirtuin Inhibitors in Human Leukemia Cells

Aberrant histone deacetylase (HDAC) activity is frequent in human leukemias. However, while classical, NAD+-independent HDACs are an established therapeutic target, the relevance of NAD+-dependent HDACs (sirtuins) in leukemia treatment remains unclear. Here, we assessed the antileukemic activity of sirtuin inhibitors and of the NAD+-lowering drug FK866, alone and in combination with traditional HDAC inhibitors. Primary leukemia cells, leukemia cell lines, healthy leukocytes and hematopoietic progenitors were treated with sirtuin inhibitors (sirtinol, cambinol, EX527) and with FK866, with or without addition of the HDAC inhibitors valproic acid, sodium butyrate, and vorinostat. Cell death was quantified by propidium iodide cell staining and subsequent flow-cytometry. Apoptosis induction was monitored by cell staining with FITC-Annexin-V/propidium iodide or with TMRE followed by flow-cytometric analysis, and by measuring caspase3/7 activity. Intracellular Bax was detected by flow-cytometry and western blotting. Cellular NAD+ levels were measured by enzymatic cycling assays. Bax was overexpressed by retroviral transduction. Bax and SIRT1 were silenced by RNA-interference. Sirtuin inhibitors and FK866 synergistically enhanced HDAC inhibitor activity in leukemia cells, but not in healthy leukocytes and hematopoietic progenitors. In leukemia cells, HDAC inhibitors were found to induce upregulation of Bax, a pro-apoptotic Bcl2 family-member whose translocation to mitochondria is normally prevented by SIRT1. As a result, leukemia cells become sensitized to sirtuin inhibitor-induced apoptosis. In conclusion, NAD+-independent HDACs and sirtuins cooperate in leukemia cells to avoid apoptosis. Combining sirtuin with HDAC inhibitors results in synergistic antileukemic activity that could be therapeutically exploited

Proceedings of the Fifth Italian Conference on Computational Linguistics CLiC-it 2018

On behalf of the Program Committee, a very warm welcome to the Fifth Italian Conference on Computational Linguistics (CLiC-­‐it 2018). This edition of the conference is held in Torino. The conference is locally organised by the University of Torino and hosted into its prestigious main lecture hall “Cavallerizza Reale”. The CLiC-­‐it conference series is an initiative of the Italian Association for Computational Linguistics (AILC) which, after five years of activity, has clearly established itself as the premier national forum for research and development in the fields of Computational Linguistics and Natural Language Processing, where leading researchers and practitioners from academia and industry meet to share their research results, experiences, and challenges

Biotecnologia dei lieviti di interesse enologico per il miglioramento della qualità dei vini

In enologia è ampiamente diffuso l’utilizzo di colture selezionate di S. cerevisiae che, se da un lato determinano un rapido avvio ed un buon controllo del processo fermentativo, dall’altro, limitando la presenza della microflora spontanea, riducono la complessità analitica e sensoriale del vino. È tuttavia possibile conferire al vino caratteristiche peculiari, garantendo al tempo stesso un regolare decorso fermentativo, attraverso l’utilizzo di starter misti controllati, costituiti da lieviti non- Saccharomyces e S. cerevisiae. A tal fine, sedici ceppi selezionati di lieviti non-Saccharomyces di interesse enologico e appartenenti a generi differenti sono stati utilizzati, su scala di laboratorio, in colture miste con S. cerevisiae in diversi rapporti di inoculo e valutati in base al loro impatto sia sulle cinetiche di fermentazione sia sulle caratteristiche analitiche dei fermentati. I risultati ottenuti hanno permesso di individuare due ceppi di lievito appartenenti alle specie Lachancea thermotolerans e Zygosaccharomyces florentinus, che nelle condizioni di coltura mista hanno determinato un miglioramento dei caratteri analitici dei fermentati. Tali colture sono state poi utilizzate in fermentazioni miste su scala di laboratorio per valutare le modalità di inoculo (coinoculo ed inoculo dopo 24 e 48 ore del S. cerevisiae) e diverse temperature di fermentazione. I risultati hanno messo in evidenza che i due fattori oggetto della sperimentazione possono influenzare in vario modo il decorso fermentativo ed il profilo analitico dei vini ottenuti. La conferma dei risultati ottenuti a livello di laboratorio è stata fatta in cantina, valutando l’attitudine enologica di ciascuno di questi due lieviti non-Saccharomyces in fermentazioni multistarter su scala semi-industriale secondo due modalità di inoculo: coinoculo ed inoculo del S. cerevisiae dopo 48 ore. I risultati ottenuti indicano che le fermentazioni miste controllate hanno determinato un più complesso profilo analitico e sensoriale dei vini, garantendo al contempo il controllo microbiologico della fermentazione

An index of 5-HT synthesis changes during early antidepressant treatment: α-[11C]methyl-l-tryptophan PET study

The antidepressant selective serotonin transporter inhibitors (SSRIs) are clinically active after a delay of several weeks. Indeed, the rapid increase of serotonin (5-HT) caused by SSRIs, stimulates the 5-HT1A autoreceptors, which exert a negative feedback on the 5-HT neurotransmission. Only when autoreceptors are desensitized, can SSRIs exert their therapeutic activity. The 5-HT1A receptor antagonist pindolol has been used to accelerate the clinical effects of antidepressant by preventing the negative feedback. Using the a-[11C]methyl-L-tryptophan/positron emission tomography (PET), the goal of the present double-blind, randomized study was to compare the changes in a-[11C]methyl-L-tryptophan trapping, an index of serotonin synthesis, in patients suffering from unipolar depression treated with the SSRI citalopram (20 mg/day) plus placebo versus patients treated with citalopram plus pindol (7.5 mg/day). PET and Hamilton depression rating scale (HDRS-17) were performed at baseline, and after 10 and 24 days of antidepressant treatment. Results show that the combination citalopram plus pindol, compared to citalopram alone shows a more rapid and greater increase of an index of 5-HT synthesis in prefrontal cortex (BA 9). This research is the first human PET study demonstrating that, after 24 days, the combination SSRIs plus pindolol produces a greater increase of the metabolism of serotonin in the prefrontal cortex, an area associated to depressive symptoms

Rilocalizzazioni industriali da Torino: rendita fondiaria e piano urbanistico

Mixed starter cultures made of Saccharomyces cerevisiae EC1118 and Schizosaccharomyces japonicus #13 were inoculated in commercial grape must, and the impact of different inoculum ratios (1:1; 1:100; 1:10,000) on growth and fermentation kinetics and on the analytical profiles of the experimental wines was here evaluated. Results obtained showed that S. japonicus #13 affects S. cerevisiae growth and fermentative capability only for S. cerevisiae/S. japonicus inoculum ratio 1:10,000. The analytical profiles of the wines produced by mixed starter cultures indicated that this non-Saccharomyces yeast modulates the concentration of malic and acetic acids and of some of the most important volatile compounds, such as β-phenyl ethanol, in an inoculum-ratio-dependent fashion. Moreover, all experimental wines obtained with S. japonicus #13 in mixed cultures reached concentrations of total polysaccharides significantly higher than those obtained with pure cultures of S. cerevisiae EC1118, and total polysaccharides increased with the increase of S. japonicus #13 cell concentration. Based on these results, S. japonicus #13 might be profitably inoculated in combination with S. cerevisiae EC1118 to enhance wine complexity and aroma and to improve wine stability by increasing the final concentration of polysaccharides

Non-saccharomyces yeasts in controlled mixed culture fermentation in winemaking: the role of metabolic interactions

One of the most important technological advances in winemaking is the control of alcoholic fermentation through the inoculation of the grape juice with selected cultures of Saccharomyces cerevisiae. In the last decade several studies revaluated the involvement of non-Saccharomyces yeasts during alcoholic fermentation and their role in the metabolic impact and aroma complexity of the final product. Indeed, non-Saccharomyces yeasts may produce high amounts of different metabolites and enzymes able to release aroma compounds from precursors present in grape must (Fernandez et al., 2000). Thus, non-Saccharomyces yeasts may influence the perceivable characteristics of the final product (Romano et al., 2003). In view of the fact that natural fermentations remain uncontrolled processes, several studies have evaluated the possibility of using controlled multistarter cultures to increase the quality of wines. The use of non-Saccharomyces wine yeasts, together with Saccharomyces strains as part of multistarter fermentations, has been proposed as a tool to take advantages of natural fermentation and maintain the control of the fermentative process (Ciani et al., 2006, [Anfang et al., 2009] and [Ciani et al., 2010]). We evaluated the oenological aptitude of non-Saccharomyces wine yeasts belonging to nine different genera. Selected non-Saccharomyces strains with a commercial S. cerevisiae strain were used to set up mixed must microfermentations estimating biomass and fermentation evolution as well as analytical profile of wines. Results of screening allowed to select some non-Saccharomyces yeasts with suitable oenological properties. The comparison between mixed and pure fermentations showed variations in the analytical profiles. The presence and permanence of non-Saccharomyces yeasts during must mixed fermentations positively influence the final product. In particular, yeast interactions frequently allowed the reduction of some negative compounds produced by non-Saccharomyces yeasts such as acetic acid, and the enhancement of favourable aroma compounds (2-phenylethanol, phenylethyl acetate, isoamyl acetate). This work is financed by Consorzio Tuscania, Firenze