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Spinal cord injury represents a great health and economic burden for patients and society, and a major challenge for both the physician and the researcher. Following a lesion, a series of anatomopathological changes (glial scar formation, neuroinflammatory reactions, and cell death) occurs, associated with the poor capability of the adult CNS to restore its connectivity owing to an environment that inhibits axonal growth and the subsequent inactivation of the growth program. Some reorganization of local neural circuits and sprouting of axons can occur, and different therapeutic approaches are being tested in the early and delayed phases of disease by preventing glial scar formation, removing inhibitory molecules, or stimulating axonal growth by trophic factors, inducing physical exercise. In addition, cell therapy may be used to replace and restore lost neural circuits or to modulate the environment, as well as different types of scaffold. Finally, gene therapy could enhance axonal growth.

Las vinificaciones realizadas consistieron en cultivos puros de Saccharomyces cerevisiae, y cultivos mixtos con inoculacion secuencial de una cepa Hanseniaspora vineae (T02/5AF) y Hanseniaspora clermontiae (A10/82F), con una cepa convencional de Saccharomyces cerevisiae. Los sistemas de vinificacion realizados con uvas Tannat fueron: semipiloto, piloto y escala industrial. Fueron identificados 51 compuestos volatiles De un total de 51 compuestos volatiles cuantificados para el total de las vinificaciones, se encontraron 16 compuestos con unidades de aromas mayores a la unidad, y por lo tanto con potencial contribucion en el perfil sensorial. La caracterizacion sensorial aromatica se realizo mediante la tecnica de mapeo proyectivo, con 48 consumidores que realizaron la evaluacion sensorial del total de los tratamientos para las tres vinificaciones. El analisis factorial multiple realizado permitio comparar los resultados quimicos y sensoriales con el fin de encontrar correlaciones. Se encontro que existio concordancia entre ambos estudios lo que confirma las diferentes tendencias aromaticas encontradas en las diferentes vinificaciones. En ambos estudios se observo que las vinificaciones semipiloto estuvieron caracterizadas por los descriptores “quimico” y “floral”, las vinificaciones piloto por el descriptor “especiado”, y las vinificaciones industriales por “frutado” y “madera”. The vinifications carried out consisted of pure cultures of Saccharomyces cerevisiae, and mixed cultures with sequential inoculation of a strain Hanseniaspora vineae (T02/5AF) and Hanseniaspora clermontiae (A10/82F), with a conventional strain of Saccharomyces cerevisiae. The vinification systems carried out with Tannat grapes were: semi-pilot, pilot and industrial scale. Of a total of 51 volatile compounds quantified for the total number of vinifications, 16 compounds were found with aroma units greater than unity, and therefore with a potential contribution to the sensory profile. Aromatic sensory characterization was performed using the projective mapping technique, with 48 consumers performing the sensory evaluation of the total treatments for the three vinifications. The multiple factorial analysis carried out allowed the chemical and sensory results to be compared in order to find correlations. It was found that there was concordance between the two studies, which confirms the different aromatic tendencies found in the different vinifications. In both studies it was observed that the semi-pilot vinifications were characterized by the descriptors "chemical" and "floral", the pilot vinifications by the descriptor "spicy", and the industrial vinifications by "fruity" and "woody".

Benzenoids are compounds associated with floral and fruity flavours in flowers, fruits and leaves and present a role in hormonal signalling in plants. These molecules are produced by the phenyl ammonia lyase pathway. However, some yeasts can also synthesize them from aromatic amino acids using an alternative pathway that remains unknown. Hanseniaspora vineae can produce benzenoids at levels up to two orders of magnitude higher than Saccharomyces species, so it is a model microorganism for studying benzenoid biosynthesis pathways in yeast. According to their genomes, several enzymes have been proposed to be involved in a mandelate pathway similar to that described for some prokaryotic cells. Among them, the ARO10 gene product could present benzoylformate decarboxylase activity. This enzyme catalyses the decarboxylation of benzoylformate into benzaldehyde at the end of the mandelate pathway in benzyl alcohol formation. Two homologous genes of ARO10 were found in the two sequenced H. vineae strains. In this study, nine other H. vineae strains were analysed to detect the presence and per cent homology of ARO10 sequences by PCR using specific primers designed for this species. Also, the copy number of the genes was estimated by quantitative PCR. To verify the relation of ARO10 with the production of benzyl alcohol during fermentation, a deletion mutant in the ARO10 gene of Saccharomyces cerevisiae was used. The two HvARO10 paralogues were analysed and compared with other α-ketoacid decarboxylases at the sequence and structural level.

The genus Hanseniaspora is the main yeast group isolated from grapes and musts. In this review, we will show some of the main results obtained in real winemaking conditions for the application of Hanseniaspora vineae, the main species of what was named the “fermentation clade” of the genus. The winemaking potential of H. vineae will be discussed in order to understand how to apply it, taking advantage at the winemaking level of its capacity to produce white wines with increased sensory complexity compared with Saccharomyces cerevisiae conventional wine yeasts. This species is evaluated in some practical aspects for its fermentation capacity at low assimilable nitrogen levels. Furthermore, it lacks H2S production and has good SO2 resistance. Other desirable characteristics are also studied, such as its ability to increase both flavor complexity with neutral grape varieties (e.g., Trebbiano, Macabeu, or Albillo) and palate due to fast cell autolysis processes. Although H. vineae shows a great capacity to produce differentiated wines, some fine-tuning adjustments are still necessary to improve the end of fermentation in highly mature white grapes by sequential inoculation with S. cerevisiae in the middle of the process. Nutrient management and mixed strain selection of the Saccharomyces pairing should be carefully defined to complete total sugar depletion.

Hanseniaspora species are the main yeasts isolated from grapes and grape musts. Regarding genetic and phenotypical characterization, especially fermentative behavior, they can be classified in two technological clusters: the fruit group and the fermentation group. Among the species belonging to the last group, Hanseniaspora osmophila and Hanseniaspora vineae have been previously isolated in spontaneous fermentations of grape must. In this work, the oenological aptitudes of the two species of the fermentation group were compared with Saccharomyces cerevisiae and the main species of the fruit group, Hanseniaspora uvarum. Both H. osmophila and H. vineae conferred a positive aroma to final wines and no sensory defects were detected. Wines fermented with H. vineae presented significantly higher concentrations of 2-phenylethyl, tryptophol and tyrosol acetates, acetoin, mevalonolactone, and benzyl alcohol compared to H. osmophila. Sensorial analysis showed increased intensity of fruity and flowery notes in wines vinificated with H. vineae. In an evolutionary context, the detoxification of alcohols through a highly acetylation capacity might explain an adaption to fermentative environments. It was concluded that, although H. vineae show close alcohol fermentation adaptations to H. osmophila, the increased activation of phenylpropanoid metabolic pathway is a particular characteristic of H. vineae within this important apiculate genus.

Friedreich ataxia (FRDA) is caused by the reduced expression of the mitochondrial protein frataxin (FXN) due to an intronic GAA trinucleotide repeat expansion in the FXN gene. Although FRDA has no cure and few treatment options, there is research dedicated to finding an agent that can curb disease progression and address symptoms as neurobehavioral deficits, muscle endurance, and heart contractile dysfunctions. Because oxidative stress and mitochondrial dysfunctions are implicated in FRDA, we demonstrated the systemic delivery of catalysts activity of gold cluster superstructures (Au8-pXs) to improve cell response to mitochondrial reactive oxygen species and thereby alleviate FRDA-related pathology in mesenchymal stem cells from patients with FRDA. We also found that systemic injection of Au8-pXs ameliorated motor function and cardiac contractility of YG8sR mouse model that recapitulates the FRDA phenotype. These effects were associated to long-term improvement of mitochondrial functions and antioxidant cell responses. We related these events to an increased expression of frataxin, which was sustained by reduced autophagy. Overall, these results encourage further optimization of Au8-pXs in experimental clinical strategies for the treatment of FRDA.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that affects both lower and upper motor neurons (MNs) in the central nervous system. ALS etiology is highly multifactorial and multifarious, and an effective treatment is still lacking. Neuroinflammation is a hallmark of ALS and could be targeted to develop new therapeutic approaches. Interestingly, the transcription factor Nurr1 has been demonstrated to have an important role in the inflammatory process in several neurological disorders, such as Parkinson's disease and multiple sclerosis. In the present paper, we demonstrate for the first time that Nurr1 expression levels are upregulated in the peripheral blood of ALS patients. Moreover, we investigated Nurr1 function in the SOD1-G93A mouse model of ALS. Nurr1 was strongly upregulated in the spinal cord during the asymptomatic and early symptomatic phases of the disease, where it promoted the expression of brain-derived neurotrophic factor mRNA and the repression of NFκB pro-inflammatory targets, such as inducible nitric oxide synthase. Therefore, we hypothesize that Nurr1 is activated in an early phase of the disease as a protective endogenous anti-inflammatory mechanism, although not sufficient to reverse disease progression. On the basis of these observations, Nurr1 could represent a potential biomarker for ALS and a promising target for future therapies., Summary: We hypothesize that the transcription factor Nurr1 is activated in the early phase of the neurodegenerative disease amylotrophic lateral sclerosis (ALS), probably as a neuroprotective endogenous mechanism. Nurr1 might represent a promising target for ALS therapy.

Summary Huntington disease (HD) is an inherited late-onset neurological disorder characterized by progressive neuronal loss and disruption of cortical and basal ganglia circuits. Cell replacement using human embryonic stem cells may offer the opportunity to repair the damaged circuits and significantly ameliorate disease conditions. Here, we showed that in-vitro-differentiated human striatal progenitors undergo maturation and integrate into host circuits upon intra-striatal transplantation in a rat model of HD. By combining graft-specific immunohistochemistry, rabies virus-mediated synaptic tracing, and ex vivo electrophysiology, we showed that grafts can extend projections to the appropriate target structures, including the globus pallidus, the subthalamic nucleus, and the substantia nigra, and receive synaptic contact from both host and graft cells with 6.6 ± 1.6 inputs cell per transplanted neuron. We have also shown that transplants elicited a significant improvement in sensory-motor tasks up to 2 months post-transplant further supporting the therapeutic potential of this approach., Highlights • hESC-derived striatal progenitors give rise to MSNs in a neurotoxin model of HD • Donor transplants extend projections to appropriate striatal target regions • Grafted cells establish synaptic contact with both donor and resident cells • Transplanted animals show improvements in HD-related sensorimotor responses, In this article, Cattaneo, Buffo, Besusso, and colleagues investigate the short-term in vivo therapeutic benefits of a cell replacement approach for Huntington disease (HD) using hESC-derived MSN progenitors. Upon intra-striatal transplantation in a neurotoxin model of HD, donor cells survive and mature in the host tissue reaching appropriate target regions, establishing synaptic contacts and mitigating lesion-dependent sensorimotor deficits.

During aging, iron accumulates in brain’s regions vulnerable to neurodegeneration: the cerebral cortex and the hippocampus. However, the mechanism of iron regulation in the brain remains scarce. Here, we demonstrated for the first time the involvement of the Hepcidin/Ferroportin1 pathway in brain iron metabolism during aging.We demonstrated the alteration of BBB integrity, that leads to increased iron permeability and deregulation of iron homeostasis during aging. We found that brain iron overload drives Hepcidin upregulation and, consequently, the inhibition of the iron exporter Ferroportin1, neuroinflammation and oxidative stress. Moreover, both in the cerebral cortex and hippocampus Ferroportin1 colocalizes with astrocytes, while the iron storage protein ferritin light-chain with neurons. This differential distribution suggests that astrocytes mediate iron shuttling and neurons are unable to metabolize it. Furthermore, we observed NCOA4-dependent ferritinophagy of ferritin heavy-chain isoforms determining the increase of light-chain enriched ferritin heteropolymers that are more efficient as iron chelators. Altogether, these data highlight the involvement of the Hepcidin/Ferroportin1 axis and NCOA4 during mice aging as a response to a higher iron influx to the brain.

The neuromuscular junction (NMJ) is at the crossroad between the nervous system (NS) and the muscle. Following neurotransmitter release from the motor neurons (MNs), muscle contraction occurs and movement is generated. Besides eliciting muscle contraction, the NMJ represents a site of chemical bidirectional interplay between nerve and muscle with the active participation of Schwann cells. Indeed, signals originating from the muscle play an important role in synapse formation, stabilization, maintenance and function, both in development and adulthood. We focus here on the contribution of the Glial cell line-Derived Neurotrophic Factor (GDNF) to these processes and to its potential role in the protection of the NMJ during neurodegeneration. Historically related to the maintenance and survival of dopaminergic neurons of the substantia nigra, GDNF also plays a fundamental role in the peripheral NS (PNS). At this level, it promotes muscle trophism and it participates to the functionality of synapses. Moreover, compared to the other neurotrophic factors, GDNF shows unique peculiarities, which make its contribution essential in neurodegenerative disorders. While describing the known structural and functional changes occurring at the NMJ during neurodegeneration, we highlight the role of GDNF in the NMJ–muscle cross-talk and we review its therapeutic potential in counteracting the degenerative process occurring in the PNS in progressive and severe diseases such as Alzheimer’s disease (AD), Amyotrophic Lateral Sclerosis (ALS) and Spinal Muscular Atrophy (SMA). We also describe functional 3D neuromuscular co-culture systems that have been recently developed as a model for studying both NMJ formation in vitro and its involvement in neuromuscular disorders.

Spinal cord injury (SCI) affects 6 million people worldwide with no available treatment. Despite research advances, the inherent poor regeneration potential of the central nervous system remains a major hurdle. Small RNAs (sRNAs) 19–33 nucleotides in length are a set of non-coding RNA molecules that regulate gene expression and have emerged as key players in regulating cellular events occurring after SCI. Here we profiled a class of sRNA known as microRNAs (miRNAs) following SCI in the cortex where the cell bodies of corticospinal motor neurons are located. We identified miR-7b-3p as a candidate target given its significant upregulation after SCI in vivo and we screened by miRWalk PTM the genes predicted to be targets of miR-7b-3p (among which we identified Wipf2, a gene regulating neurite extension). Moreover, 16 genes, involved in neural regeneration and potential miR-7b-3p targets, were found to be downregulated in the cortex following SCI. We also analysed miR-7b-3p function during cortical neuron development in vitro: we observed that the overexpression of miR-7b-3p was important (1) to maintain neurons in a more immature and, likely, plastic neuronal developmental phase and (2) to contrast the apoptotic pathway; however, in normal conditions it did not affect the Wipf2 expression. On the contrary, the overexpression of miR-7b-3p upon in vitro oxidative stress condition (mimicking the SCI environment) significantly reduced the expression level of Wipf2, as observed in vivo, confirming it as a direct miR-7b-3p target. Overall, these data suggest a dual role of miR-7b-3p: (i) the induction of a more plastic neuronal condition/phase, possibly at the expense of the axon growth, (ii) the neuroprotective role exerted through the inhibition of the apoptotic cascade. Increasing the miR-7b-3p levels in case of SCI could reactivate in adult neurons silenced developmental programmes, supporting at the same time the survival of the axotomised neurons.

Benzenoid-derived metabolites act as precursors for a wide variety of products involved in essential metabolic roles in eukaryotic cells. They are synthesized in plants and some fungi through the phenylalanine ammonia lyase (PAL) and tyrosine ammonia lyase (TAL) pathways. Ascomycete yeasts and animals both lack the capacity for PAL/TAL pathways, and metabolic reactions leading to benzenoid synthesis in these organisms have remained incompletely known for decades. Here, we show genomic, transcriptomic, and metabolomic evidence that yeasts use a mandelate pathway to synthesize benzenoids, with some similarities to pathways used by bacteria. We conducted feeding experiments using a synthetic fermentation medium that contained either 13C-phenylalanine or 13C-tyrosine, and, using methylbenzoylphosphonate (MBP) to inhibit benzoylformate decarboxylase, we were able to accumulate intracellular intermediates in the yeast Hanseniaspora vineae. To further confirm this pathway, we tested in separate fermentation experiments three mutants with deletions in the key genes putatively proposed to form benzenoids (Saccharomyces cerevisiaearo10Δ, dld1Δ, and dld2Δ strains). Our results elucidate the mechanism of benzenoid synthesis in yeast through phenylpyruvate linked with the mandelate pathway to produce benzyl alcohol and 4-hydroxybenzaldehyde from the aromatic amino acids phenylalanine and tyrosine, as well as sugars. These results provide an explanation for the origin of the benzoquinone ring, 4-hydroxybenzoate, and suggest that Aro10p has benzoylformate and 4-hydroxybenzoylformate decarboxylase functions in yeast. IMPORTANCE We present here evidence of the existence of the mandelate pathway in yeast for the synthesis of benzenoids. The link between phenylpyruvate- and 4-hydroxyphenlypyruvate-derived compounds with the corresponding synthesis of benzaldehydes through benzoylformate decarboxylation is demonstrated. Hanseniaspora vineae was used in these studies because of its capacity to produce benzenoid derivatives at a level 2 orders of magnitude higher than that produced by Saccharomyces. Contrary to what was hypothesized, neither β-oxidation derivatives nor 4-coumaric acid is an intermediate in the synthesis of yeast benzenoids. Our results might offer an answer to the long-standing question of the origin of 4-hydroxybenzoate for the synthesis of Q10 in humans.

Mixed strain dynamics are still not well or easily monitored although recently molecular identification methods have improved our knowledge. This study used a chromogenic differential plating medium that allows the discrimination of four of the main selected biofuel strains that are currently under development for ethanol production from cellulosic hydrolysates. Complete fermentation of hexoses and xylose was obtained with a yeast consortium composed of Spathaspora passalidarum, Scheffersomyces stipitis, Candida akabanensis and Saccharomyces cerevisiae. The results showed that C.akabanensis excessively dominated consortium balance. Reducing its inoculum from 33 to 4.8% improved population strain balance and fermentation efficiency. Comparison of the consortia with single strain fermentations showed that it optimize sugar consumption and ethanol yields. This simple and cheap method also has advantages compared with molecular methods, as the yeast strains do not need to be genetically marked and identified cell proportions are probably active in the fermentation system as compared to DNA determination methods.

Mitochondria play a central role in a plethora of processes related to the maintenance of cellular homeostasis and genomic integrity. They contribute to preserving the optimal functioning of cells and protecting them from potential DNA damage which could result in mutations and disease. However, perturbations of the system due to senescence or environmental factors induce alterations of the physiological balance and lead to the impairment of mitochondrial functions. After the description of the crucial roles of mitochondria for cell survival and activity, the core of this review focuses on the “mitochondrial switch” which occurs at the onset of neuronal degeneration. We dissect the pathways related to mitochondrial dysfunctions which are shared among the most frequent or disabling neurodegenerative diseases such as Alzheimer’s, Parkinson’s, and Huntington’s, Amyotrophic Lateral Sclerosis, and Spinal Muscular Atrophy. Can mitochondrial dysfunctions (affecting their morphology and activities) represent the early event eliciting the shift towards pathological neurobiological processes? Can mitochondria represent a common target against neurodegeneration? We also review here the drugs that target mitochondria in neurodegenerative diseases.

Vitis vinifera flowers and grape fruits are one of the most interesting ecosystem niches for native yeasts development. There are more than a 100 yeast species and millions of strains that participate and contribute to design the microbial terroir. The wine terroir concept is understood when grape and wine micro-regions were delimited by different quality characteristics after humans had been growing vines for more than 10,000 years. Environmental conditions, such as climate, soil composition, water management, winds and air quality, altitude, fauna and flora and microbes, are considered part of the "terroir" and contribute to a unique wine style. If "low input winemaking" strategies are applied, the terroir effect will be expected to be more authentic in terms of quality differentiation. Interestingly, the role of the microbial flora associated with vines was very little study until recently when new genetic technologies for massive species identification were developed. These biotechnologies allowed following their environmental changes and their effect in shaping the microbial profiles of different wine regions. In this chapter we explain the interesting positive effects on flavor diversity and wine quality obtained by using "friendly" native yeasts that allowed the microbial terroir flora to participate and contribute during fermentation.

Schinus molle L. (Anacardiaceae) is an evergreen tree native to South America and has been introduced into North and South Africa and the Mediterranean region. The mature berries are known as an alternative to pepper by their pungency and collected for essential oil production to substitute black pepper in perfumery. Several investigators have examined the physicochemical properties and chemical composition of the essential oil, but the release of bound volatile compounds in S. molle berries from the corresponding glycosides is presented here for the first time. The carotenoid content of mature berries was also studied over two successive ripening periods. Ten carotenoids were identified by HPLC-MS/MS: lutein, phytoene, β-cryptoxanthin, phytofluene, β-carotene, 9-Z-β-carotene, β-cryptoxanthin-C12:0, β-cryptoxanthin-C14:0, β-cryptoxanthin-C16:0 and lycopene. This research is the first to characterise the carotenoids in molle berries and their degradation products (norisoprenoids) in the "free" and glycosylated volatile fruit fractions. The detection of many of these glycosidically bound volatile compounds in berries should be considered to have a sensory contribution, which might differentiate the volatile profile. In addition, our results could explain, through the flavour complexity found in both the "free" and glycosylated fractions, the reported traditional use of molle berries as a pepper substitute. For comparison purposes, the essential oil from mature berries obtained by hydrodistillation was also studied.

Phenolic compounds can affect the sensory characteristics of quality wines, yet the chemical modifications induced by yeasts on phenolic compounds have been studied only to a limited extent. Forty-nine non-Saccharomyces yeast strains belonging to different genera were screened for their effects on color and fermentation capability in a synthetic must supplemented with anthocyanins from Tannat grapes. Total anthocyanins, color intensity, hue, and total polyphenol index were significantly affected (p

New players are emerging in the game of peripheral and central nervous system injury since their physiopathological mechanisms remain partially elusive. These mechanisms are characterized by several molecules whose activation and/or modification following a trauma is often controlled at transcriptional level. In this scenario, microRNAs (miRNAs/miRs) have been identified as main actors in coordinating important molecular pathways in nerve or spinal cord injury (SCI). miRNAs are small non-coding RNAs whose functionality at network level is now emerging as a new level of complexity. Indeed they can act as an organized network to provide a precise control of several biological processes. Here we describe the functional synergy of some miRNAs in case of SCI and peripheral damage. In particular we show how several small RNAs can cooperate in influencing simultaneously the molecular pathways orchestrating axon regeneration, inflammation, apoptosis and remyelination. We report about the networks for which miRNA-target bindings have been experimentally demonstrated or inferred based on target prediction data: in both cases, the connection between one miRNA and its downstream pathway is derived from a validated observation or is predicted from the literature. Hence, we discuss the importance of miRNAs in some pathological processes focusing on their functional structure as participating in a cooperative and/or convergence network.

A pineapple vinification process was conducted through inoculated and spontaneous fermentation to develop a process suitable for a quality beverage during two successive vintages in Huambo, Angola. Wines obtained with the conventional Saccharomyces cerevisiae strain, were analysed by gas chromatography, and a total of 61 volatile constituents were detected in the volatile fraction and 18 as glycosidically bound aroma compounds. Concentration levels of carbonyl and sulphur compounds were in agreement with the limited information reported about pineapple fruits of other regions. We report, for the first time in pineapple wines, the presence of significant concentrations of lactones, ketones, terpenes, norisoprenoids and a variety of volatile phenols. Eight native yeast strains were isolated from spontaneous batches. Further single-strain fermentations allowed us to characterise their suitability for commercial fermentation. Three native strains (Hanseniaspora opuntiae, H. uvarum and Meyerozyma guilliermondii) were selected with sensory potential to ferment pineapple fruits with increased flavour diversity. Results obtained here contribute to a better understanding of quality fermentation alternatives of this tropical fruit in subtropical regions.

Previously, we reported the first oxygen partial pressure (Po2) measurements in the brain of awake mice, by performing two-photon phosphorescence lifetime microscopy at micrometer resolution (Lyons et al., 2016). However, this study disregarded that imaging through a cranial window lowers brain temperature, an effect capable of affecting cerebral blood flow, the properties of the oxygen sensors and thus Po2 measurements. Here, we show that in awake mice chronically implanted with a glass window over a craniotomy or a thinned-skull surface, the postsurgical decrease of brain temperature recovers within a few days. However, upon imaging with a water immersion objective at room temperature, brain temperature decreases by ~2–3°C, causing drops in resting capillary blood flow, capillary Po2, hemoglobin saturation, and tissue Po2. These adverse effects are corrected by heating the immersion objective or avoided by imaging through a dry air objective, thereby revealing the physiological values of brain oxygenation., eLife, 8, ISSN:2050-084X

The c-Jun NH2-terminal protein kinase (JNK) is a Janus-faced kinase, which, in the nervous system, plays important roles in a broad range of physiological and pathological processes. Three genes, encoding for 10 JNK isoforms, have been identified: jnk1, jnk2, and jnk3. In the developing spinal cord, JNK proteins control neuronal polarity, axon growth/pathfinding, and programmed cell death; in adulthood they can drive degeneration and regeneration, after pathological insults. Indeed, recent studies have highlighted a role for JNK in motor neuron (MN) diseases, such as amyotrophic lateral sclerosis and spinal muscular atrophy. In this review we discuss how JNK-dependent signaling regulates apparently contradictory functions in the spinal cord, in both the developmental and adult stages. In addition, we examine the evidence that the specific targeting of JNK signaling pathway may represent a promising therapeutic strategy for the treatment of MN diseases.

Flavour and some compounds associated with wine colour are known to be yeast strain-dependent. These metabolites are important for the sensory quality of wines, studies searching for increase aroma and color are a key area today in winemaking. The aim of this work was to study the oenological potential of the two main strains of Hanseniaspora vineae,native to Uruguay to better understand their successful application at winery level. It is known that these strains contribute with extracellular proteases and β-glucosidase enzyme activities that might increase cell lysis and flavor depending in grape varieties. Application and nutrient management of the process of these strains in production of white wines (Chardonnay, Macabeo and Petit Manseng) and red wine Tannat are discussed. Wines were evaluated to determine the volatile compounds composition and their effect compared to conventional processes. Low production of short and medium chain fatty acids and ethyl esters, and high production of acetate esters and isoprenoids are found compared to S. cerevisiaestrains. The most outstanding characteristic of the species H. vineaewas the production of benzenoids, phenylpropanoids and acetate esters. This behavior was reflected in the sensory evaluation, where all the fermentations performed with H. vineaewere considered superior compared to Saccharomyces cerevisiaewine strains.

In this chapter we present results of red grape maceration methods, with practical application, and their impact on wine yeast biotechnologies. It is well known that Saccharomyces cerevisiae wine yeasts produce flavor compounds at different concentrations as a function of grape must treatments and fermentation conditions. Factors such as grape variety, micronutrients and vitamins, yeast cell concentration, temperature, redox situation, and nitrogen composition of must have been studied. In red winemaking, skin contact and grape maceration processes dramatically affect yeast development due to increased yeast diversity and microaerobic conditions of the system. In addition, the greater number of variables in red wine processes, such as tank size and barrel fermentations, makes research approaches more complex compared with white wine. As experimental designs are more difficult in these conditions, studies of the impact of S. cerevisiae metabolism on red color and phenolic compounds has been addressed to a very limited extent.

Tannat (Vitis vinifera) is the most cultivated grapevine variety in Uruguay for the production of high quality red wines. Its berries have unusually high levels of polyphenolic compounds (anthocyanins and tannins), producing wines with intense purple colour and high antioxidant properties. Remarkably, more than 40% of its tannins are galloylated, which determines a greater antioxidant power. Technologies of massive sequencing allow the characterization of genomic variants between different clutivars. The Tannat genome was sequenced with a 134X coverage using the Illumina technology, and was annotated using transcriptomes (RNA-Seq) of different berry tissues. When comparing the genomes of Tannat with Pinot Noir PN40024 (reference genome) we found an expansion of the gene families related to the biosynthesis of polyphenols. A search base on the recently reported epicatechin galloyl transferase (ECGT) from tea leaves determined five putative genes encoding the ECGT in Tannat. Genetic analysis of one of the transcription factor that regulates the anthocyanin synthesis during berry ripening, VvMYBA1, shows the presence of Gret1 retrotransposon in one of the VvMYBA1 alleles in the Tannat clones analysed. This work makes original contributions about the molecular bases of the biosynthesis of anthocyanins and tannins during the development of the flagship grape of Uruguay.

Vinifications were conducted using pure cultures of Saccharomyces cerevisiae, and mixed cultures by sequential inoculation of a Hanseniaspora vineae (T02/05F) and Hanseniaspora clermontiae (A10/82F), with a Saccharomyces cerevisiae conventional strain. The vinification systems applied to Tannat grapes were defined: semipilot, pilot and industrial scale. Fifty-one volatile compounds were identified in all the vinifications, sixteen of them were above the aroma threshold values and potentially contributed to the final sensory profiles. A sensory characterization of aroma was carried out using the projective mapping technique, with forty-eight consumers who made evaluation of the three vinifications. Multiple factorial analysis was used to compare the chemical and sensory data to find correlations. The results obtained from both methodologies were coincident, confirming the aromatic tendencies found in the different vinifications. Both studies demonstrated that wines obtained by semipilot scale was characterized by descriptors associated with “chemical” and “floral”; those from pilot vinification by the “spicy” descriptor; while industrial vinification produced wines described as “fruit” and “wood”.

Epilepsy is a complex clinical condition characterized by repeated spontaneous seizures. Seizures have been linked to multiple drivers including DNA damage accumulation. Investigation of epilepsy physiopathology in humans imposes ethical and practical limitations, for this reason model systems are mostly preferred. Among animal models, mouse mutants are particularly valuable since they allow conjoint behavioral, organismal, and genetic analyses. Along with this, since aging has been associated with higher frequency of seizures, prematurely aging mice, simulating human progeroid diseases, offer a further useful modeling element as they recapitulate aging over a short time-window. Here we report on a mouse mutant with progeroid traits that displays repeated spontaneous seizures. Mutant mice were produced by reducing the expression of the gene Ft1 (AKTIP in humans). In vitro, AKTIP/Ft1 depletion causes telomere aberrations, DNA damage, and cell senescence. AKTIP/Ft1 interacts with lamins, which control nuclear architecture and DNA function. Premature aging defects of Ft1 mutant mice include skeletal alterations and lipodystrophy. The epileptic behavior of Ft1 mutant animals was age and sex linked. Seizures were observed in 18 mutant mice (23.6% of aged ≥ 21 weeks), at an average frequency of 2.33 events/mouse. Time distribution of seizures indicated non-random enrichment of seizures over the follow-up period, with 75% of seizures happening in consecutive weeks. The analysis of epileptic brains did not reveal overt brain morphological alterations or severe neurodegeneration, however, Ft1 reduction induced expression of the inflammatory markers IL-6 and TGF-β. Importantly, Ft1 mutant mice with concomitant genetic reduction of the guardian of the genome, p53, showed no seizures or inflammatory marker activation, implicating the DNA damage response into these phenotypes. This work adds insights into the connection among DNA damage, brain function, and aging. In addition, it further underscores the importance of model organisms for studying specific phenotypes, along with permitting the analysis of genetic interactions at the organismal level. Published version

Spinal muscular atrophy (SMA) is a recessive autosomal neuromuscular disease, due to homozygous mutations or deletions in the telomeric survival motoneuron gene 1 (SMN1). SMA is characterized by motor impairment, muscle atrophy, and premature death following motor neuron (MN) degeneration. Emerging evidence suggests that dysregulation of autophagy contributes to MN degeneration. We here investigated the role of autophagy in the SMNdelta7 mouse model of SMA II (intermediate form of the disease) which leads to motor impairment by postnatal day 5 (P5) and to death by P13. We first showed by immunoblots that Beclin 1 and LC3-II expression levels increased in the lumbar spinal cord of the SMA pups. Electron microscopy and immunofluorescence studies confirmed that autophagic markers were enhanced in the ventral horn of SMA pups. To clarify the role of autophagy, we administered intracerebroventricularly (at P3) either an autophagy inhibitor (3-methyladenine, 3-MA), or an autophagy inducer (rapamycin) in SMA pups. Motor behavior was assessed daily with different tests: tail suspension, righting reflex, and hindlimb suspension tests. 3-MA significantly improved motor performance, extended the lifespan, and delayed MN death in lumbar spinal cord (10372.36 ± 2716 MNs) compared to control-group (5148.38 ± 94 MNs). Inhibition of autophagy by 3-MA suppressed autophagosome formation, reduced the apoptotic activation (cleaved caspase-3 and Bcl2) and the appearance of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive neurons, underlining that apoptosis and autophagy pathways are intricately intertwined. Therefore, autophagy is likely involved in MN death in SMA II, suggesting that it might represent a promising target for delaying the progression of SMA in humans as well. The project was supported by grants of the Associazione Girotondo Onlus (Trivero, Italy), the Smarathon Association (Milan, Italy), and by a grant of the Italian Ministry of Health (Project RF-2009-1475235). AP was the recipient of a fellowship (3 years) from the ‘’Master and Back” project 2009 by the Regione Autonoma della Sardegna (Cagliari, Italy).

In several grape varieties, the dominating aryl alkyl alcohols found are the volatile group of phenylpropanoid-related compounds, such as glycosylated benzyl and 2-phenylethyl alcohol, which contribute to wine with floral and fruity aromas after being hydrolysed during fermentation. Saccharomyces cerevisiae is largely recognized as the main agent in grape must fermentation, but yeast strains belonging to other genera, including Hanseniaspora, are known to predominate during the first stages of alcoholic fermentation. Although non-Saccharomyces yeast strains have a well-recognized genetic diversity, understanding of their impact on wine flavour richness is still emerging. In this study, 11 Hansenisapora vineae strains were used to ferment a chemically defined simil-grape fermentation medium, resembling the nutrient composition of grape juice but devoid of grape-derived secondary metabolites. GC-MS analysis was performed to determine volatile compounds in the produced wines. Our results showed that benzyl alcohol, benzyl acetate and 2-phenylethyl acetate are significantly synthesized by H. vineae strains. Levels of these compounds found in fermentations with 11 H. vineae different strains were one or two orders of magnitude higher than those measured in fermentations with a known S. cerevisiae wine strain. The implications for winemaking in response to the negative correlation of benzyl alcohol, benzyl acetate and 2-phenylethyl acetate production with yeast assimilable nitrogen concentrations are discussed. Copyright © 2016 John Wiley & Sons, Ltd.

During fermentation, Saccharomyces cerevisiae releases into the medium secondary metabolic products, such as acetaldehyde, able to react with anthocyanins, producing more stable derived pigments. However, very limited reports are found about non-Saccharomyces effects on grape fermentation. In this study, six non-Saccharomyces yeast strains, belonging to the genera Metschnikowia and Hanseniaspora, were screened for their effect on red wine colour and wine-making capacity under pure culture conditions and mixed with Saccharomyces. An artificial red grape must was prepared, containing a phenolic extract of Tannat grapes that allows monitoring changes of key phenol parameters during fermentation, but without skin solids in the medium. When fermented in pure cultures, S. cerevisiae produced higher concentrations of acetaldehyde and vitisin B (acetaldehyde reaction-dependent) compared to M. pulcherrima M00/09G, Hanseniaspora guillermondii T06/09G, H. opuntiae T06/01G, H. vineae T02/05F and H. clermontiae (A10/82Fand C10/54F). However, co-fermentation of H. vineae and H. clermontiae with S. cerevisiae resulted in a significantly higher concentration of acetaldehyde compared with the pure S. cerevisiae control. HPLC-DAD-MS analysis confirmed an increased formation of vitisin B in co-fermentation treatments when compared to pure Saccharomyces fermentation, suggesting the key role of acetaldehyde. Copyright © 2016 John Wiley & Sons, Ltd.

Hanseniaspora species can be isolated from grapes and grape musts, but after the initiation of spontaneous fermentation, they are displaced by Saccharomyces cerevisiae. Hanseniaspora vineae is particularly valuable since this species improves the flavour of wines and has an increased capacity to ferment relative to other apiculate yeasts. Genomic, transcriptomic, and metabolomic studies in H. vineae have enhanced our understanding of its potential utility within the wine industry. Here, we compared gene sequences of 12 glycolytic and fermentation pathway enzymes from five sequenced Hanseniaspora species and S. cerevisiae with the corresponding enzymes encoded within the two sequenced H. vineae genomes. Increased levels of protein similarity were observed for enzymes of H. vineae and S. cerevisiae, relative to the remaining Hanseniaspora species. Key differences between H. vineae and H. uvarum pyruvate kinase enzymes might explain observed differences in fermentative capacity. Further, the presence of eight putative alcohol dehydrogenases, invertase activity, and sulfite tolerance are distinctive characteristics of H. vineae, compared to other Hanseniaspora species. The definition of two clear technological groups within the Hanseniaspora genus is discussed within the slow and fast evolution concept framework previously discovered in these apiculate yeasts.

Pachyrhizus ahipa is an Andean leguminous plant which has purple-pigmented edible roots. The content and distribution of the phenolic compounds in the skins and flesh of P. ahipa roots as well as their antioxidant capacity were studied. Total phenols, flavonoids, tannins and anthocyanins were quantified and the effect of the extraction solvent on the maximum absorbance wavelength (λvis-max) was measured. The ATR-FTIR spectrum and stability of the purified anthocyanin extracts were analyzed. Individual anthocyanins were partially characterized using high-performance liquid chromatography-mass spectrometry and absorbance spectra. Ahipa skins had much more phenols (3.25 mg gallic acid equivalents/g), flavonoids (3.0 mg catechin equivalents/g) and tannins (4.6 mg tannic acid equivalents/g) than the flesh, which was reflected in a higher antioxidant capacity. Changing the extraction solvent from acidified methanol to acidified water shifted the λvis-max 16 nm lower, suggesting copigmentation reactions. Anthocyanin in the skins was >8.7 content in the flesh, and the chromatographic analysis showed at least 6 different anthocyanins with identical retention times for skins and flesh, including delphinidin, pelargonidin and cyanidin as aglycons. The absorbance spectra showed that two of them are acylated structures. Pigments were stable for at least 180 min at pH 1 and 25 °C, but they were affected by increased pH and temperature. These anthocyanins may be a potential source of natural pigments. Fil: Dini, Cecilia. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos; Argentina Fil: Zaro, Maria Jose. Facultad de Ciencias Agrarias y Forestales, Universidad Nacional de la Plata; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos; Argentina Fil: Rolny, Nadia Soledad. Universidad Nacional de La Plata. Facultad de Ciencias Agrarias y Forestales; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos; Argentina Fil: Caputo, Maricel. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Química. Laboratorio de Estudio de Compuestos Orgánicos; Argentina Fil: Boido, E.. Universidad de la República; Uruguay Fil: Dellacassa, E.. Universidad de la República; Uruguay Fil: Viña, Sonia Zulma. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Agrarias y Forestales; Argentina

Current efforts to improve muscle performance are focused on muscle trophism via inhibition of the myostatin pathway: however they have been unsuccessful in the clinic to date. In this study, a novel protein has been created by combining the soluble activin receptor, a strong myostatin inhibitor, to the C-terminal agrin nLG3 domain (ActR-Fc-nLG3) involved in the development and maintenance of neuromuscular junctions. Both domains are connected via the constant region of an Igg1 monoclonal antibody. Surprisingly, young male mice treated with ActR-Fc-nLG3 showed a remarkably increased endurance in the rotarod test, significantly longer than the single domain compounds ActR-Fc and Fc-nLG3 treated animals. This increase in endurance was accompanied by only a moderate increase in body weights and wet muscle weights of ActR-Fc-nLG3 treated animals and were lower than expected. The myostatin inhibitor ActR-Fc induced, as expected, a highly significant increase in body and muscle weights compared to control animals and ActR-Fc-nLG3 treated animals. Moreover, the prolonged endurance effect was not observed when ActR-Fc and Fc-nLG3 were dosed simultaneously as a mixture and the body and muscle weights of these animals were very similar to ActR-Fc treated animals, indicating that both domains need to be on one molecule. Muscle morphology induced by ActR-Fc-nLG3 did not appear to be changed however, close examination of the neuromuscular junction showed significantly increased acetylcholine receptor surface area for ActR-Fc-nLG3 treated animals compared to controls. This result is consistent with published observations that endurance training in rats increased acetylcholine receptor quantity at neuromuscular junctions and provide evidence that improving nerve-muscle interaction could be an important factor for sustaining long term muscle activity.

Apiculate yeasts of the genus Hanseniaspora/Kloeckera are the main species present on mature grapes and play a significant role at the beginning of fermentation, producing enzymes and aroma compounds that expand the diversity of wine color and flavor. Ten species of the genus Hanseniaspora have been recovered from grapes and are associated in two groups: H. valbyensis, H. guilliermondii, H. uvarum, H. opuntiae, H. thailandica, H. meyeri, and H. clermontiae; and H. vineae, H. osmophila, and H. occidentalis. This review focuses on the application of some strains belonging to this genus in co-fermentation with Saccharomyces cerevisiae that demonstrates their positive contribution to winemaking. Some consistent results have shown more intense flavors and complex, full-bodied wines, compared with wines produced by the use of S. cerevisiae alone. Recent genetic and physiologic studies have improved the knowledge of the Hanseniaspora/Kloeckera species. Significant increases in acetyl esters, benzenoids, and sesquiterpene flavor compounds, and relative decreases in alcohols and acids have been reported, due to different fermentation pathways compared to conventional wine yeasts.

Hanseniaspora is the main genus of the apiculate yeast group that represents approximately 70% of the grape-associated microflora. Hanseniaspora vineae is emerging as a promising species for quality wine production compared to other non-Saccharomyces species. Wines produced by H. vineae with Saccharomyces cerevisiae consistently exhibit more intense fruity flavors and complexity than wines produced by S. cerevisiae alone. In this work, genome sequencing, assembling, and phylogenetic analysis of two strains of H. vineae showed that it is a member of the Saccharomyces complex and it diverged before the whole-genome duplication (WGD) event from this clade. Specific flavor gene duplications and absences were identified in the H. vineae genome compared to 14 fully sequenced industrial S. cerevisiae genomes. The increased formation of 2-phenylethyl acetate and phenylpropanoids such as 2-phenylethyl and benzyl alcohols might be explained by gene duplications of H. vineae aromatic amino acid aminotransferases (ARO8 and ARO9) and phenylpyruvate decarboxylases (ARO10). Transcriptome and aroma profiles under fermentation conditions confirmed these genes were highly expressed at the beginning of stationary phase coupled to the production of their related compounds. The extremely high level of acetate esters produced by H. vineae compared to that by S. cerevisiae is consistent with the identification of six novel proteins with alcohol acetyltransferase (AATase) domains. The absence of the branched-chain amino acid transaminases (BAT2) and acyl coenzyme A (acyl-CoA)/ethanol O-acyltransferases (EEB1) genes correlates with H. vineae's reduced production of branched-chain higher alcohols, fatty acids, and ethyl esters, respectively. Our study provides sustenance for understanding and potentially utilizing genes that determine fermentation aromas. Fil: Giorello, Facundo. Universidad de la Republica; Uruguay Fil: Valera, Maria Jose. Universidad de la Republica; Uruguay Fil: Martín, Valentina. Universidad de la Republica; Uruguay. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Parada, Andres. Universidad Austral de Chile; Chile Fil: Salzman, Valentina. Instituto Pasteur de Montevideo; Uruguay Fil: Camesasca, Laura. Instituto de Investigaciones Biológicas Clemente Estable; Uruguay Fil: Fariña, Laura. Universidad de la República; Uruguay Fil: Boido, Eduardo. Universidad de la República; Uruguay Fil: Medina, Karina. Universidad de la República; Uruguay Fil: Dellacassa, Eduardo. Universidad de la República; Uruguay Fil: Berna, Luisa. Universidad de la República; Uruguay Fil: Aguilar, Pablo Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Mas, Albert. Universitat Rovira I Virgili; España Fil: Gaggero, Carina. Instituto de Investigaciones Biológicas Clemente Estable; Uruguay Fil: Carrau, Francisco. Universidad de la República; Uruguay

Amyotrophic Lateral Sclerosis (ALS) is a neural disorder gradually leading to paralysis of the whole body. Alterations in superoxide dismutase SOD1 gene have been linked with several variants of familial ALS. Here, we investigated a transgenic (Tg) cloned swine model expressing the human pathological hSOD1G93A allele. As in patients, these Tg pigs transmitted the disease to the progeny with an autosomal dominant trait and showed ALS onset from about 27 months of age. Post mortem analysis revealed motor neuron (MN) degeneration, gliosis and hSOD1 protein aggregates in brainstem and spinal cord. Severe skeletal muscle pathology including necrosis and inflammation was observed at the end stage, as well. Remarkably, as in human patients, these Tg pigs showed a quite long presymptomatic phase in which gradually increasing amounts of TDP-43 were detected in peripheral blood mononuclear cells. Thus, this transgenic swine model opens the unique opportunity to investigate ALS biomarkers even before disease onset other than testing novel drugs and possible medical devices.

Tannat is a red variety of Vitis vinifera that has become the major variety for the production of premium red wines in Uruguay. Due to its small cultivation around the world, research on the viticulture and enology of this variety is still necessary to improve wine quality. In this context, the aim of the present work was to characterize the aroma profile of Uruguayan Tannat wines using chemical and sensory methodologies. The volatile composition of ten Uruguayan Tannat wines, sold in the international market, was studied by gas-chromatography (GC–MS). Sixty two volatile compounds were identified using GC–MS, being alcohols and esters the most abundant compounds. Only few volatile compounds were found at concentrations higher than their odor threshold in all samples. Sensory characterization of wine aroma was characterized by a panel of wine professionals using projective mapping. Red fruits, fruity, dry fruits, and woody were the main descriptors used for describing similarities and differences in the aroma profile of the wines. Projective mapping sorted samples into four main groups. Partial least square regression (PLSR) enabled to explain many of the most important sensory descriptors (woody, earthy, phenolic, sulfur, chemical and microbiological) through volatile composition.

Starting from a series of arylazoenamine derivatives, shown to be selectively and potently active against the bovine viral diarrhea virus (BVDV), we developed a hierarchical combined experimental/molecular modeling strategy to explore the drug leads for the BVDV RNA-dependent RNA polymerase. Accordingly, BVDV mutants resistant to lead compounds in our series were isolated, and the mutant residues on the viral molecular target, the RNA-dependent RNA polymerase, were identified. Docking procedures upon previously identified pharmacophoric constraints and actual mutational data were carried out, and the binding affinity of all active compounds for the RdRp was estimated. Given the excellent agreement between in silico and in vitro data, this procedure is currently being employed in the design a new series of more selective and potent BVDV inhibitors.

The benefits of tonsillectomy in IgA nephropathy (IgAN) are still debated. Tonsillectomy may remove pathogen sources and reduce the mucosal associated lymphoid tissue (MALT), limiting degalactosylated IgA1 (deGal-IgA1) production, which is considered to be the initiating pathogenetic event leading to IgA glomerular deposition. In the European network VALIGA, 62/1147 IgAN patients underwent tonsillectomy (TxIgAN). In a cross-sectional study 15 of these patients were tested and compared to 45 non-tonsillectomized IgAN (no-TxIgAN) and healthy controls (HC) regarding levels of deGal-IgA1, and markers of innate immunity and oxidative stress, including toll-like receptors (TLR)2, 3, 4 and 9 mRNAs, proteasome (PS) and immunoproteasome (iPS) mRNAs in peripheral blood mononuclear cells (PBMC), and advanced oxidation protein products (AOPP). Levels of deGal-IgA1 were lower in TxIgAN than in no-TxIgAN (p = 0.015), but higher than in HC (p = 0.003). TLR mRNAs were more expressed in TxIgAN than in HC (TLR4, p = 0.021; TLR9, p = 0.027), and higher in TxIgAN than in no-TxIgAN (p ≤ 0.001 for TLR2, 4, 9). A switch from PS to iPS was detected in PBMC of TxIgAN in comparison to HC and it was higher than in no-TxIgAN [large multifunctional peptidase (LMP)2/β1, p = 0.039; LPM7/β5, p < 0.0001]. The levels of AOPP were significantly higher in TxIgAN than HC (p < 0.001) and no-TxIgAN (p = 0.033). In conclusion, the activation of innate immunity via TLRs and ubiquitin–proteasome pathways and the pro-oxidative milieu were not affected by tonsillectomy, even though the levels of aberrantly galactosylated IgA1 were lower in patients with IgAN who had tonsillectomy. The residual hyperactivation of innate immunity in tonsillectomized patients may result from extra-tonsillar MALT.

Eighty-five arylazoenamines, characterized by different types of aryl and basic moieties, have been synthesized and evaluated in cell-based assays for cytotoxicity and antiviral activity against a panel of ten RNA and DNA viruses. The most commonly affected viruses were, in decreasing order, CVB-2, RSV, BVDV, YFV, and Sb-1; the remaining viruses were either not affected (HIV-1, VSV, and VV) or susceptible only to a very few compounds (Reo-1 and HSV-1). Thirty-five compounds exhibited high activity, with EC(50) in the range 0.8-10 microM, and other 28 compounds had EC(50) between 11 and 30 microM, thus indicating that the arylazoenamine molecular pattern is an interesting novel pharmacophore for antiviral agents against ssRNA viruses. Moreover, some compounds (as 28, 32, 42, and 53) appear of high interest, being devoid of toxicity on the human MT-4 cells (CC(50)>100 microM). A ligand-based computational approach was employed to identify highly predictive pharmacophore models for the most frequently affected viruses CVB-2, RSV, and BVDV. These models should allow the design of second generation of more potent inhibitors of these human and veterinary pathogens.

Mutations occurring in the CFTR gene, encoding for the cystic fibrosis transmembrane conductance regulator chloride channel, cause cystic fibrosis (CF). Mutations belonging to class II, such as DeltaPhe508, give rise to a protein with both a defective maturation and altered channel gating. Mutations belonging to class III, such as G551D and G1349D, cause only a gating defect. We have previously identified antihypertensive 1,4-dihydropyridines (DHPs), a class of drugs that block voltage-dependent Ca(2+) channels, as effective potentiators of CFTR gating, able to correct the defective activity of CFTR mutants (Mol Pharmacol 68:1736-1746, 2005). However, optimization of potency for CFTR versus Ca(2+) channels is required to design selective compounds for CFTR pharmacotherapy. In the present study, we have established DHP structure-activity relationship for both CFTR potentiation and Ca(2+) channel inhibition using cell-based assays for both types of channels. A panel of 333 felodipine analogs was studied to understand the effect of various substitutions and modifications in the DHP scaffold. Our results show that alkyl substitutions at the para position of the 4-phenyl ring lead to compounds with very low activity on Ca(2+) channels and strong effect as potentiators on the DeltaPhe508, G551D, and G1349D CFTR mutants.

Brain cholesterol biosynthesis and cholesterol levels are reduced in mouse models of Huntington's disease (HD), suggesting that locally synthesized, newly formed cholesterol is less available to neurons. This may be detrimental for neuronal function, especially given that locally synthesized cholesterol is implicated in synapse integrity and remodeling. Here, we used biodegradable and biocompatible polymeric nanoparticles (NPs) modified with glycopeptides (g7) and loaded with cholesterol (g7-NPs-Chol), which per se is not blood-brain barrier (BBB) permeable, to obtain high-rate cholesterol delivery into the brain after intraperitoneal injection in HD mice. We report that g7-NPs, in contrast to unmodified NPs, efficiently crossed the BBB and localized in glial and neuronal cells in different brain regions. We also found that repeated systemic delivery of g7-NPs-Chol rescued synaptic and cognitive dysfunction and partially improved global activity in HD mice. These results demonstrate that cholesterol supplementation to the HD brain reverses functional alterations associated with HD and highlight the potential of this new drug-administration route to the diseased brain. Synopsis: Cholesterol in brain is largely derived by local synthesis. One affected pathway in Huntington's disease (HD) implicates that reduced production and/or availability of brain cholesterol may be detrimental for neuronal function. Polymeric nanoparticles (NPs) loaded with cholesterol, modified with glycopeptides g7 (g7-NPs-Chol) to cross the BBB after systemic injection, have been used to deliver cholesterol to the brain of HD mice. These NPs, applied for the first time to a brain disorder, are made of PLGA, which is approved by FDA in various drug delivery systems in humans. Systemic administration of g7-NPs-Chol (i) rescued synaptic communication in striatal medium-sized spiny neurons, (ii) prevented cognitive decline, and (iii) restored the levels of proteins that compose the synaptic machinery in HD mice. g7-NPs or cholesterol itself did not induce inflammatory response in the periphery, where almost all g7-NPs are localized. Cholesterol in brain is largely derived by local synthesis. One affected pathway in Huntington's disease (HD) implicates that reduced production and/or availability of brain cholesterol may be detrimental for neuronal function.

Spinal muscular atrophy (SMA) is a severe neuromuscular disease, the most common in infancy, and the third one among young people under 18 years. The major pathological landmark of SMA is a selective degeneration of lower motor neurons, resulting in progressive skeletal muscle denervation, atrophy, and paralysis. Recently, it has been shown that specific or general changes in the activity of ribonucleoprotein containing micro RNAs (miRNAs) play a role in the development of SMA. Additionally miRNA-206 has been shown to be required for efficient regeneration of neuromuscular synapses after acute nerve injury in an ALS mouse model. Therefore, we correlated the morphology and the architecture of the neuromuscular junctions (NMJs) of quadriceps, a muscle affected in the early stage of the disease, with the expression levels of miRNA-206 in a mouse model of intermediate SMA (SMAII), one of the most frequently used experimental model. Our results showed a decrease in the percentage of type II fibers, an increase in atrophic muscle fibers and a remarkable accumulation of neurofilament (NF) in the pre-synaptic terminal of the NMJs in the quadriceps of SMAII mice. Furthermore, molecular investigation showed a direct link between miRNA-206-HDAC4-FGFBP1, and in particular, a strong up-regulation of this pathway in the late phase of the disease. We propose that miRNA-206 is activated as survival endogenous mechanism, although not sufficient to rescue the integrity of motor neurons. We speculate that early modulation of miRNA-206 expression might delay SMA neurodegenerative pathway and that miRNA-206 could be an innovative, still relatively unexplored, therapeutic target for SMA.

Synapsins (Syn I, Syn II, and Syn III) are a family of synaptic vesicle phosphoproteins regulating synaptic transmission and plasticity. SYN1/2 genes have been identified as major epilepsy susceptibility genes in humans and synapsin I/II/III triple knockout (TKO) mice are epileptic. However, excitatory and inhibitory synaptic transmission and short-term plasticity have never been analyzed in intact neuronal circuits of TKO mice. To clarify the generation and expression of the epileptic phenotype, we performed patch-clamp recordings in the CA1 region of acute hippocampal slices from 1-month-old presymptomatic and 6-month-old epileptic TKO mice and age-matched controls. We found a strong imbalance between basal glutamatergic and γ-aminobutyric acid (GABA)ergic transmission with increased evoked excitatory postsynaptic current and impaired evoked inhibitory postsynaptic current amplitude. This imbalance was accompanied by a parallel derangement of short-term plasticity paradigms, with enhanced facilitation of glutamatergic transmission in the presymptomatic phase and milder depression of inhibitory synapses in the symptomatic phase. Interestingly, a lower tonic GABA(A) current due to the impaired GABA release is responsible for the more depolarized resting potential found in TKO CA1 neurons, which makes them more susceptible to fire. All these changes preceded the appearance of epilepsy, indicating that the distinct changes in excitatory and inhibitory transmission due to the absence of Syns initiate the epileptogenic process.

Intrinsic homeostasis enables neuronal circuits to maintain activity levels within an appropriate range by modulating neuronal voltage-gated conductances, but the signalling pathways involved in this process are largely unknown. We characterized the process of intrinsic homeostasis induced by sustained electrical activity in cultured hippocampal neurons based on the activation of the Repressor Element-1 Silencing Transcription Factor/Neuron-Restrictive Silencer Factor (REST/NRSF). We showed that 4-aminopyridine-induced hyperactivity enhances the expression of REST/NRSF, which in turn, reduces the expression of voltage-gated Na+ channels, thereby decreasing the neuronal Na+ current density. This mechanism plays an important role in the downregulation of the firing activity at the single-cell level, re-establishing a physiological spiking activity in the entire neuronal network. Conversely, interfering with REST/NRSF expression impaired this homeostatic response. Our results identify REST/NRSF as a critical factor linking neuronal activity to the activation of intrinsic homeostasis and restoring a physiological level of activity in the entire neuronal network.

The volatile compounds of 18 yerba mate (YM) commercial brands from Argentina, Brazil, P.araguay and Uruguay were extracted using a Likens–Nickerson simultaneous distillation–extraction (SDE) apparatus, and analyzed by gas chromatography–mass spectrometry (GC–MS). Results showed that a total of 35 major volatile compounds were identified by GC–MS; among them, norisoprenoid compounds. A principal component analysis (PCA) was carried out in order to ascertain the degree of differentiation between samples and which compounds were involved. The results showed three main groups of YM corresponding to the places where they are commercialized. The first group (samples from Paraguay and Argentina) correlated positively with 6-methyl-5-hepten-2-one, octanal and 1-octanol. The second group (YM marketed in Uruguay but produced in Brazil) correlated positively with α- and β-ionone, linalool, nerol, α-terpineol, ( Z )-linalool oxide and the aliphatic hydrocarbons undecane, dodecane and heptadecane. The third group (YM produced and marketed in Brazil) correlated positively with 2,4-heptadienal isomers, ( E )-linalool oxide and β-cyclocitral. Through a preference test, one commercial brand was selected to be analyzed by gas chromatography–olfactometry (GC–O). Sixteen odor active compounds were perceived by GC–O analysis. It seems that the norisoprenoid compounds and components produced by fatty acid degradation play a vital role in the special aroma of yerba mate.

Synapsins (SynI, SynII, SynIII) are a multigene family of synaptic vesicle phosphoproteins implicated in the regulation of synaptic transmission and plasticity. Synapsin I, II, I/II and I/II/III knockout mice are epileptic and SYN1/2 genes have been identified as major epilepsy susceptibility genes in humans. We analyzed cortico-hippocampal hyperexcitability and epileptiform activity induced by 4-aminopyridine (4AP) in acute slices from presymptomatic (3-weeks-old) and epileptic (1-year-old) SynI/II/III triple knockout (TKO) mice and aged-matched triple wild type (TWT) controls and assessed the effect of the synaptic vesicle-targeted antiepileptic drug levetiracetam (LEV) in reverting the epileptic phenotype. Both fast and slow interictal (I-IC) activity and ictal (IC) seizures were observed in both genotypes. The incidence of fast I-IC was higher in presymptomatic TKO slices, while frequency and latency of I-IC events were similar in both genotypes. The major age and genotype effects were observed in IC activity, that was much more pronounced in 3-weeks-old TKO and persisted with age, while it disappeared from 1-year-old TWT slices. LEV virtually suppressed fast I-IC and IC discharges from 3-weeks-old TWT slices, while it only ameliorated fast I-IC and IC activity in TKO slices. Analysis of I-IC events in patch-clamped CA1 pyramidal neurons revealed that LEV increased the inhibitory/excitatory ratio of I-IC activity in both genotypes. The lower LEV potency in TKO slices of both ages was associated with a decreased expression of SV2A, a synaptic vesicle protein acting as LEV receptor, in cortex and hippocampus. The results demonstrate that deletion of synapsin genes is associated with a higher propensity to 4AP-induced epileptic paroxysms that precedes the onset of epilepsy and consolidates with age. LEV ameliorates such hyperexcitability by enhancing the inhibition/excitation ratio, although the effect is hindered in TKO slices due to the concomitant decrease of its receptor SV2A.

Benzyl alcohol and other benzenoid-derived metabolites of particular importance in plants confer floral and fruity flavors to wines. Among the volatile aroma components in Vitis vinifera grape varieties, benzyl alcohol is present in its free and glycosylated forms. These compounds are considered to originate from grapes only and not from fermentative processes. We have found increased levels of benzyl alcohol in red Tannat wine compared to that in grape juice, suggesting de novo formation of this metabolite during vinification. In this work, we show that benzyl alcohol, benzaldehyde, p-hydroxybenzaldehyde, and p-hydroxybenzyl alcohol are synthesized de novo in the absence of grape-derived precursors by Hanseniaspora vineae. Levels of benzyl alcohol produced by 11 different H. vineae strains were 20-200 times higher than those measured in fermentations with Saccharomyces cerevisiae strains. These results show that H. vineae contributes to flavor diversity by increasing grape variety aroma concentration in a chemically defined medium. Feeding experiments with phenylalanine, tryptophan, tyrosine, p-aminobenzoic acid, and ammonium in an artificial medium were tested to evaluate the effect of these compounds either as precursors or as potential pathway regulators for the formation of benzenoid-derived aromas. Genomic analysis shows that the phenylalanine ammonia-lyase (PAL) and tyrosine ammonia lyase (TAL) pathways, used by plants to generate benzyl alcohols from aromatic amino acids, are absent in the H. vineae genome. Consequently, alternative pathways derived from chorismate with mandelate as an intermediate are discussed. Fil: Martín, Valentina. Universidad de la República; Uruguay. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Giorello, Facundo. Universidad de la República; Uruguay Fil: Fariña, Laura. Universidad de la República; Uruguay. Instituto de Investigaciones Biológicas "Clemente Estable"; Uruguay Fil: Minteguiaga, Manuel. Universidad de la República; Uruguay Fil: Salzman, Valentina. Instituto Pasteur de Montevideo; Uruguay. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Boido, Eduardo. Universidad de la República; Uruguay Fil: Aguilar, Pablo Sebastián. Instituto Pasteur de Montevideo; Uruguay. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Gaggero, Carina. Instituto de Investigaciones Biológicas "Clemente Estable"; Uruguay Fil: Dellacassa, Eduardo. Universidad de la República; Uruguay Fil: Mas, Albert. Universitat Rovira I Virgili; España Fil: Carrau, Francisco. Universidad de la República; Uruguay

Amyloid-β 1-42 accumulation is the major pathogenetic event in Alzheimer's disease (AD), believed to be responsible for synaptic dysfunction and neuronal cell death. However, the physiologic activity of Aβ peptides remains elusive: Aβ might not only play a toxic role, but also act as a functional signaling intermediate. We recently reported that Aβ1-42 promotes BACE1 transcription through the activation of the JNK-c-jun pathway. Here, we show that the Aβ1-42-mediated increase in BACE1 expression is accompanied by a decrease in ubiquitin C-terminal hydrolase L1 (Uch-L1) expression and activity in different cellular models such as neuroblastoma SH-SY5Y as well as NT(2) neuronal cells. We also found that the increase in BACE1 and the decrease in Uch-L1 are related events and depend on NF-κB pathway; thus, Aβ1-42 is able to activate NF-κB pathway and the pretreatment with a pharmacological inhibitor, able to block the nuclear translocation of the transactivating unit p65, almost completely prevents both the decrease in Uch-L1 and the increase in BACE1 expression. In addition, the decrease in Uch-L1 activity interferes with the lysosomal degradation of BACE1, as demonstrated by the decrease in Cathepsin D activity and the partial accumulation of BACE1 in lysosomes after Aβ1-42 treatment as well after Uch-L1 inhibition. In support of the in vitro data, we observed low protein levels of Uch-L1 associated with high protein levels of BACE1 in sporadic AD brains. Our data suggest that Uch-L1 could be an attractive target for the development of new therapeutic approaches for AD.

The objective measure of grape quality is an important requirement to enhance wine quality. Particularly, the quantitation of glycosylated aroma compounds could be a useful index for winemakers. In the present work, we evaluated the bound compounds profile on Tannat grapes, and the potential of near infrared (NIR) spectroscopy coupled to chemometric techniques to determine the concentration of the different glycosylated aroma compounds present in this grape variety. The results showed phenols and norisoprenoids as the quantitative most important group of compounds, representing almost 80 % of the total glycosidic components present in Tannat grapes. The glycosidic form of ethyl-β-(4-hydroxy-3-methoxy-phenyl)-propionate was identified for the first time in grapes. The results suggested NIR technique as a screening tool for a fast analysis of the grape aroma potential. The application of such methodology would be ideal for fast grape screening purposes, contributing to the winemaking decisions, classification of the fruit, and perhaps maturity monitoring.