The autism-associated Meis2 gene is necessary for cardiac baroreflex regulation in mice.

Recent understanding of Autism Spectrum Disorder (ASD) showed that peripheral primary mechanosensitive neurons involved in touch sensation and central neurons affected in ASD share transcriptional regulators. Mutant mice for ASD-associated transcription factors exhibit impaired primary tactile perception and restoring those genes specifically in primary sensory neurons rescues some of the anxiety-like behavior and social interaction defects. Interestingly, peripheral mechanosensitive sensory neurons also project to internal organs including the cardiovascular system, and an imbalance of the cardio-vascular sympathovagal regulation is evidenced in ASD and intellectual disability. ASD patients have decreased vagal tone, suggesting dysfunction of sensory neurons involved in cardio-vascular sensing. In light of our previous finding that the ASD-associated Meis2 gene is necessary for normal touch neuron development and function, we investigated here if its inactivation in mouse peripheral sensory neurons also affects cardio-vascular sympathovagal regulation and baroreflex. Combining echocardiography, pharmacological challenge, blood pressure monitoring, and heart rate variability analysis, we found that Meis2 mutant mice exhibited a blunted vagal response independently of any apparent cardiac malformation. These results suggest that defects in primary sensory neurons with mechanosensitive identity could participate in the imbalanced cardio-vascular sympathovagal tone found in ASD patients, reinforcing current hypotheses on the role of primary sensory neurons in the etiology of ASD.

The impact of grape processing and carbonic maceration on the microbiota of early stages of winemaking.

AIMS: The work investigates the impact of grapes processing at the beginning of winemaking on the composition of microbiota during the oenological fermentations and on the composition of obtained wines. METHODS AND RESULTS: The experiments were conducted in a biodynamic winery to exclude interference due to microbial starters. Three winemaking protocols, with different pre-fermentative management of grapes, were followed by plate count and next generation sequencing on 16S for bacteria and internal transcribed spacer sequencing (ITS) for yeast. Chemical and sensory characterization of wine was performed. The grape processing influenced the evolution of microbiota (especially lactic and acetic acid bacteria) and the fermentation rate. The highest biodiversity was observed in the experiment carried out with whole grapes and carbonic maceration, with the presence of bacterial groups not usually found in winemaking (Bacteroidales, Clostridiales, Oscillospira). The different microbiotas influenced the organic acid profile of wines, the content of biogenic ammines and the perception of organoleptic descriptors linked to the vine cultivar (Syrah). CONCLUSIONS: Carbonic maceration impacts on the evolution of the microbiota and the wine features. The absence of addition of starters and sulphur dioxide would seem to be correlated with the high microbial biodiversity. SIGNIFICANCE AND IMPACT OF THE STUDY: Carbonic maceration is a traditional winemaking practice, today there are difficulties in its managing because the anaerobiosis stimulates spoilage micro-organisms. The work elucidates the reasons of these difficulties and identified some microbial groups rarely associated with winemaking. The ratio of ethanol accumulation along with physical management of grapes and the supply of oxygen during the early stages of winemaking are powerful instruments of oenological variability, able to offer new possibilities to winemakers in order to defining the quality of red wines.

Suspect screening of glycoalkaloids in plant extracts using neutral loss - High resolution mass spectrometry.

Alkaloids represent a very large and chemically inhomogeneous group of nitrogen compounds isolated from different natural sources. They have long been studied due to their specific toxicological characteristics, as some of them are suspected of having very dangerous properties, but there has still been little investigation of the existence of possible glycoside precursors in plant products. An analytical method based on a suspect screening approach using liquid chromatography coupled with high resolution tandem mass spectrometry in neutral loss (NL) mode was used for the tentative identification of alkaloid precursors. This NL approach applied to 15 plant extracts made it possible to tentatively identify 41 glycosidic precursors (4 monopentosides, 3 monodeoxyhexosides, 24 monohexosides, 1 pentoside-hexoside and 9 dihexosides) in 6 of them, belonging to the piperidine (21), pyrrolizidine (9), isoquinoline (7), indole (3) and tropane (1) alkaloid groups. This work confirms the promising investigative ability of a suspect screening approach in this complex field of study.

Glycosylated simple phenolic profiling of food tannins using high resolution mass spectrometry (Q-Orbitrap).

Tannins are polyphenolic compounds extensively present in plants and used by food industry as processing aids. Due to the heterogeneity of plant sources, actions involved in food processing and tannin commercial costs can be different. In the last years different approaches aimed at correctly identifying the tannin botanical origin have been developed, in order to satisfy the industry's request to verify product labels. This work aimed to define the glycosidic simple phenolic profile of a large selection of monovarietal commercial tannins of different origin, using a high-resolution untargeted approach. Using accurate mass, isotopic pattern and MS/MS fragmentation, 167 precursors, 89 as monoglycosylated and 78 as diglycosylated derivatives were tentatively identified in tannins, validating the untargeted approach with 3 custom-synthesized glycosidic precursors. Almost all tannin botanical varieties were shown to be characterised by a specific glycosylated phenolic profile, providing possible tools for tannin classification in the case of glycosylphenol standard availability.

Non-targeted glycosidic profiling of international wines using neutral loss-high resolution mass spectrometry.

Many metabolites naturally occur as glycosides, since sugar moieties can be crucial for their biological activity and increase their water solubility. In the plant kingdom they may occur as glycosides or sugar esters, depending on precursor chemical structure, and in wine they have traditionally attracted attention due to their organoleptic properties, such as astringency and bitterness, and because they affect the colour and aroma of wines. A new approach directed at detailed description of glycosides in a large selection of monovarietal wines (8 samples each of Pinot Blanc, Muller Thurgau, Riesling, Traminer, Merlot, Pinot Noir and Cabernet Sauvignon) was developed by combining high performance liquid chromatography with high resolution tandem mass spectrometry. Analytical separation was performed on an Accucore™ Polar Premium LC column, while mass analysis was performed in negative ion mode with an non-targeted screening approach, using a Full MS/AIF/NL dd-MS2 experiment at a resolving power of 140,000 FWHM. Over 280 glycoside-like compounds were detected, of which 133 (including low-molecular weight phenols, flavonoids and monoterpenols) were tentatively identified in the form of pentose (6), deoxyhexose (17), hexose (73), hexose-pentose (16), hexose-deoxyhexose (7), dihexose (5) and hexose ester (9) derivatives. It was not possible to univocally define the corresponding chemical structure for the remaining 149 glycosides. Non-parametric statistical analysis showed it was possible to well characterise the glycosylated profile of all red and Traminer wines, while the identified glycosides were almost entirely lacking in Pinot Blanc, Riesling and Muller Thurgau wines. Also Tukey's Honestly Significant Difference test (p < 0.05) and Principal Component Analysis confirmed that it was possible to almost entirely distinguish the selected red wines from each other according to their glycosylated profile.

Application of ozone during grape drying for the production of straw wine. Effects on the microbiota and compositive profile of grapes.

AIMS: Straw wine is a highly valuable oenological production, affected by relevant economical losses due to proliferation of spoilage micro-organisms during drying grapes, after being harvested. In this work, ozone was evaluated as a tool to preserve grapes during drying, in terms of both qualitative and quantitative changes induced in the epiphytic microflora. In addition, the alteration exerted by ozone on grapes' chemical composition was analysed. METHODS AND RESULTS: Grapes from four vine varieties were treated with ozone produced by a cold plasma generator during the entire drying period (6 weeks). The microflora was quantified weekly by plate counts and characterized by 454-pyrosequencing, and was compared with identical, untreated grape samples. At the end of drying, an extensive chemical characterization of the whole mass of grapes by FT-IR and GC-MS was performed. Ozone counteracted the growth of microflora by reducing the microbial population up to 3 log units. From the qualitative point of view, ozone reduced the incidence of spoilage micro-organisms, such as Acetobacter and Botrytis cinerea, among the microbiota of grapes. CONCLUSIONS: The statistical analysis discriminates grape samples based on the residual epiphytic microflora at the end of drying and not on their chemical composition. These evidences confirmed that ozone did control spoilage micro-organisms without altering the volatile profile of grapes. Chemical analysis revealed that untreated grapes are less suitable for winemaking owing to the deprivation of some valuable compounds during the microbial proliferation. SIGNIFICANCE AND IMPACT OF THE STUDY: Ozone is an emerging tool in winemaking, recently applied for controlling spoilage micro-organisms in winery. The present study describes a new application of ozone as an effective and safe alternative to chemical preservatives which are actually involved in the control of microbial alterations of grapes.

Targeted and untargeted characterisation of free and glycosylated simple phenols in cocoa beans using high resolution-tandem mass spectrometry (Q-Orbitrap).

Free simple phenols have positive effects on health and influence the organoleptic profile of cocoa products, contributing towards defining their aroma and nutritional properties. Glycosidically bound simple phenols can be hydrolysed during the production phase to the corresponding free forms, and thus potentially contribute to the final sensory profile. In this work, 60 samples of Forastero cocoa beans from all over the world were analysed, combining on-line solid phase extraction (SPE) clean-up with ultra-high performance liquid chromatography (UHPLC), coupled with high resolution mass spectrometry. Operating in negative ion mode and with a heated electrospray, 62 simple phenols were measured, of which four were glucosidic precursors, with quantification limits ranging from 0.04 to 40mgkg-1, calibration R2 of 0.99 for over 93% of compounds, and precision (R.S.D.%) always lower than 12%. On the basis of accurate mass, isotope pattern and MS/MS spectrum, 32 monoglycosylated simple phenols such as hexoside and pentoside precursors, and 14 diglycosylated simple phenols such as hexoside-hexoside, hexoside-pentoside and pentoside-hexoside precursors, were tentatively identified. The untargeted approach was validated using 3 glucosidic precursors synthesized by an external supplier. Honestly Significant Difference Tukey's test (p<0.05) and Discriminant Analysis showed it was possible to distinguish the geographical origin of cocoa beans. In particular, the absolute free phenol profile made it possible to characterise 4 out of 5 production macro-areas well, while an untargeted approach based on the ionisation profiles of glycosylated forms allowed complete characterisation of all the 5 macro-areas.

Free and glycosylated simple phenol profiling in Apulian Italian wines.

Free simple phenols have a significant role in defining the sensory and nutritional characteristics of wines, affecting the organoleptic profile and having positive effects on health, but glycosidically bound phenols can also be hydrolysed during the winemaking process, releasing the corresponding volatile compounds and making a possible contribution to the final sensory profile. In this work, application of on-line SPE liquid chromatography-high resolution mass spectrometry, operating in negative polarity with heated electrospray, allowed to detect over eighty free and glycosylated simple phenols in Primitivo di Manduria and Negroamaro wines. Sixty-one phenols, four of which phenolic glucosidic precursors, were quantified as having quantification limits ranging from 0.001 to 0.1µgmL(-1), calibration R(2) of 0.99 for over 92% of compounds, and precision (R.S.D.%) always lower than 12%. Twenty-four simple phenolic precursors were tentatively identified as hexoside, pentoside and hexoside-hexoside derivatives, on the basis of accurate mass, isotopic pattern and MS/MS fragmentation.

A reappraisal of ICU and long-term outcome of allogeneic hematopoietic stem cell transplantation patients and reassessment of prognosis factors: results of a 5-year cohort study (2009-2013).

Epidemiology and prognosis of complications related to allogeneic hematopoietic stem cell transplant (HSCT) recipients requiring admission to intensive care unit (ICU) have not been reassessed precisely in the past few years. We performed a retrospective single-center study on 318 consecutive HSCT patients (2009-2013), analyzing outcome and factors prognostic of ICU admission. Among these patients, 73 were admitted to the ICU. In all, 32 patients (40.3%) died in ICU, 46 at hospital discharge (63%) and 61 (83.6%) 1 year later. Survivors had a significantly lower sequential organ failure assessment (SOFA) score, serum lactate and bilirubin upon ICU admission. Catecholamine support, mechanical ventilation (MV) and/or renal replacement therapy during ICU stay, a delayed organ support and an active graft versus host disease (GvHD) significantly worsen the outcome. By multivariate analysis, the worsening of SOFA score from days 1 to 3, the need for MV and the occurrence of an active GvHD were predictive of mortality. In conclusion, the incidence of HSCT-related complications requiring an admission to an ICU was at 22%, with an ICU mortality rate of 44%, and 84% 1 year later. A degradation of SOFA score at day 3 of ICU, need of MV and occurrence of an active GvHD are main predictive factors of mortality.

Evaluation of the oenological suitability of grapes grown using biodynamic agriculture: the case of a bad vintage.

AIMS: We compare the evolution of the microbiota of grapes grown following conventional or biodynamic protocols during the final stage of ripening and wine fermentation in a year characterized by adverse climatic conditions. METHODS AND RESULTS: The observations were made in a vineyard subdivided into two parts, cultivated using a biodynamic and traditional approach in a year which saw a combination of adverse events in terms of weather, creating the conditions for extensive proliferation of vine pests. The biodynamic approach was severely tested, as agrochemicals were not used and vine pests were counteracted with moderate use of copper, sulphur and plant extracts and with intensive use of agronomical practices aimed at improving the health of the vines. Agronomic, microbiological and chemical testing showed that the response of the vineyard cultivated using a biodynamic approach was comparable or better to that of vines cultivated using the conventional method. CONCLUSIONS: The work suggests that biodynamic cultivation of the grapevine may be sustainable even in difficult conditions, representing an interesting alternative to traditional vine-growing approaches. SIGNIFICANCE AND IMPACT OF THE STUDY: This theme is topical and of interest to winemakers and consumers today, but is not easy to study due to the difficulties in finding vineyards with homogeneous characteristics, cultivated using different agronomical protocols. The particular climatic conditions observed in 2014 made this year a rare model, making it possible to verify the applicability of biodynamics to vine growing. The strict experimental plan gave results particularly useful for understanding the features of grape microbiota in a biodynamic context.