Chitosan-based hydrogel to support the paracrine activity of mesenchymal stem cells in spinal cord injury treatment

Abstract Advanced therapies which combine cells with biomaterial-based carriers are recognized as an emerging and powerful method to treat challenging diseases, such as spinal cord injury (SCI). By enhancing transplanted cell survival and grafting, biomimetic hydrogels can be properly engineered to encapsulate cells and locate them at the injured site in a minimally invasive way. In this work, chitosan (CS) based hydrogels were developed to host mesenchymal stem cells (MSCs), since their paracrine action can therapeutically enhance the SC regeneration, limiting the formation of a glial scar and reducing cell death at the injured site. An injectable and highly permeable CS-based hydrogel was fabricated having a rapid gelation upon temperature increase from 0 to 37 °C. CS was selected as former material both for its high biocompatibility that guarantees the proper environment for MSCs survival and for its ability to provide anti-inflammatory and anti-oxidant cues. MSCs were mixed with the hydrogel solution prior to gelation. MSC viability was not affected by the CS hydrogel and encapsulated MSCs were able to release MSC-vesicles as well as to maintain their anti-oxidant features. Finally, preliminary in vivo tests on SCI mice revealed good handling of the CS solution loading MSCs during implantation and high encapsulated MSCs survival after 7 days

Molecular diversity within clones of cv. Tannat (Vitis vinifera)

DNAs from 9 clones of cv. Tannat (Vitis vinifera) were analyzed at 89 microsatellite loci. Only one, VMCNg 1d12, showed a differential pattern that separated the clones in two groups. The statistical analysis of concentrations for aroma compounds from microvinifications also resulted in the same two groupings of clones. Many analyzed microsatellite loci amplified only one allele, implying that Tannat is a highly homozygous variety. For a given set of 15 microsatellites the level of homozygosity was 53 % for Tannat, in contrast to 6 % for Pinot, 20 % for both Cabernet Franc and Chardonnay and 33 % for Cabernet Sauvignon. We provide molecular data for Tannat, originating from southwestern France and nowadays becoming the emblematic cultivar of Uruguayan fine red wines. We also report a correlation between aroma-related compounds and molecular markers within clones of a cultivar.

Impact on Tannat wines aroma produced by different yeast using three vinification systems

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”

Oenological attributes of the yeast Hanseniaspora vineaeand its application for white and red winemaking

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

Estudio de precursores aromáticos en clones comerciales de Tannat y seguimiento de su evolución durante la maduración

Resumen La variedad Tannat desde el punto de vista del aroma, resulta una variedad neutra pero capaz de aportar precursores aromáticos que son de interés en la elaboración de vinos de guarda. Los principales precursores aromáticos presentes son compuestos glicosidados y carotenoides. Durante 3 vendimias consecutivas se estudió el contenido de precursores aromáticos de 8 clones comerciales de Tannat (ENTAV-INRA: 398, 399, 472, 474, 475, 717, 794 Y 944). En base a los resultados obtenidos se seleccionaron los clones 717 y 474 por su alta y baja performance en la producción de precursores glicosidados. En la siguiente vendimia se estudió el comportamiento de precursores glicosidados (SPE-GC-MS) y carotenoides (HPLC-DAD) en los clones seleccionados durante 4 estadíos de madurez de la uva, donde se identificaron 14 xantofilas y 1 caroteno. Se encontraron diferencias significativas en el contenido de carotenoides entre los dos clones estudiados a lo largo del período de maduración. Mientras que su degradación no presentó diferencias significativas. Conjuntamente se identificaron 10 norisoprenoides glicosidados en la uva de los clones estudiados pero el contenido de estos no presentó diferencias significativas entre los clones durante está vendimia. Las diferencias entre los clones 717 y 474 son tales que no se expresan todos los años teniendo variabilidad con el año y donde el clima juega un rol fundamental

Chemical characterization and enological potential of less frequent red grape Uruguayan varieties by study secondary metabolites

Study of secondary grape metabolites, such as polyphenol compounds (anthocyanins, flavonols, flavan-3-ols), volatile compounds present in grapes in both free and glycoside forms (monoterpenols, norisoprenoids, benzenoids), allowed us to characterize grape varieties and to suggest the best winemaking practices to maximize their enological potential. The Vitis viníferas studied in the present work are less frequent red grape varieties cultivated in small vineyards in southern Uruguay (Montevideo and Canelones Provinces): Ancellota, Aspiran Bouschet (syn. Lacryma Christi), Marselan (Grenache × Cabernet Sauvignon), Arinarnoa (Tannat × Cabernet Sauvignon), Egiodola (Abouriou × Tinta Negra Mole) and Caladoc (Malbec × Grenache)

Estrategias combinadas para la obtención de un vino Tannat con un contenido moderado de alcohol

Resumen En este trabajo se buscó a través de la combinación de 2 estrategias reducir el contenido de alcohol en vinos de la variedad Tannat. La primera estrategia consistió en el screening dentro de clones uruguayos de Tannat Con este fin se muestrearon durante tres vendimias consecutivas 10 clones uruguayos de Tannat y se utilizó como referencia el clon francés 717 en viñedos. De los clones estudiados 4 presentaron durante la vendimia 2023 un contenido significativamente inferior de alcohol que el clon 717, manteniendo los componentes responsables de su calidad sensorial (aromas glicosidados, antocianos totales). Mientras que la segunda estrategia fue realizar un screening de levaduras no-Saccharomyces nativas aisladas de viñedos uruguayos para utilizar en conjunto con la cepa Saccharomyces cerevisiae Lalvin BM4x4 Lalleman (utilizada como referencia para vinificar vino Tannat). Se estudió el comportamiento de 4 géneros de levaduras (Candida zemplinina, Metschnikowia pulcherrima, Metschnikowia fructicola y Hanseniaspora uvarum), en microvinificaciones en símil vino. Estas estrategias fueron seleccionada por que además de permitir la reducción de alcohol en vinos Tannat en forma conjunta le aportan una identidad propia de nuestro país

Transplantation of clinical-grade human neural stem cells reduces neuroinflammation, prolongs survival and delays disease progression in the SOD1 rats.

Abstract Stem cells are emerging as a therapeutic option for incurable diseases, such as Amyotrophic Lateral Sclerosis (ALS). However, critical issues are related to their origin as well as to the need to deepen our knowledge of the therapeutic actions exerted by these cells. Here, we investigate the therapeutic potential of clinical-grade human neural stem cells (hNSCs) that have been successfully used in a recently concluded phase I clinical trial for ALS patients (NCT01640067). The hNSCs were transplanted bilaterally into the anterior horns of the lumbar spinal cord (four grafts each, segments L3–L4) of superoxide dismutase 1 G93A transgenic rats (SOD1 rats) at the symptomatic stage. Controls included untreated SOD1 rats (CTRL) and those treated with HBSS (HBSS). Motor symptoms and histological hallmarks of the disease were evaluated at three progressive time points: 15 and 40 days after transplant (DAT), and end stage. Animals were treated by transient immunosuppression (for 15 days, starting at time of transplantation). Under these conditions, hNSCs integrated extensively within the cord, differentiated into neural phenotypes and migrated rostro-caudally, up to 3.77 ± 0.63 cm from the injection site. The transplanted cells delayed decreases in body weight and deterioration of motor performance in the SOD1 rats. At 40DAT, the anterior horns at L3–L4 revealed a higher density of motoneurons and fewer activated astroglial and microglial cells. Accordingly, the overall survival of transplanted rats was significantly enhanced with no rejection of hNSCs observed. We demonstrated that the beneficial effects observed after stem cell transplantation arises from multiple events that counteract several aspects of the disease, a crucial feature for multifactorial diseases, such as ALS. The combination of therapeutic approaches that target different pathogenic mechanisms of the disorder, including pharmacology, molecular therapy and cell transplantation, will increase the chances of a clinically successful therapy for ALS

Perivascular spaces in the brain:anatomy, physiology and pathology

Perivascular spaces include a variety of passageways around arterioles, capillaries and venules in the brain, along which a range of substances can move. Although perivascular spaces were first identified over 150 years ago, they have come to prominence recently owing to advances in knowledge of their roles in clearance of interstitial fluid and waste from the brain, particularly during sleep, and in the pathogenesis of small vessel disease, Alzheimer disease and other neurodegenerative and inflammatory disorders. Experimental advances have facilitated in vivo studies of perivascular space function in intact rodent models during wakefulness and sleep, and MRI in humans has enabled perivascular space morphology to be related to cognitive function, vascular risk factors, vascular and neurodegenerative brain lesions, sleep patterns and cerebral haemodynamics. Many questions about perivascular spaces remain, but what is now clear is that normal perivascular space function is important for maintaining brain health. Here, we review perivascular space anatomy, physiology and pathology, particularly as seen with MRI in humans, and consider translation from models to humans to highlight knowns, unknowns, controversies and clinical relevance

De-Novo Assembly and Analysis of the Heterozygous Triploid Genome of the Wine Spoilage Yeast Dekkera bruxellensis AWRI1499

Despite its industrial importance, the yeast species Dekkera (Brettanomyces) bruxellensis has remained poorly understood at the genetic level. In this study we describe whole genome sequencing and analysis for a prevalent wine spoilage strain, AWRI1499. The 12.7 Mb assembly, consisting of 324 contigs in 99 scaffolds (super-contigs) at 26-fold coverage, exhibits a relatively high density of single nucleotide polymorphisms (SNPs). Haplotype sampling for 1.2% of open reading frames suggested that the D. bruxellensis AWRI1499 genome is comprised of a moderately heterozygous diploid genome, in combination with a divergent haploid genome. Gene content analysis revealed enrichment in membrane proteins, particularly transporters, along with oxidoreductase enzymes. Availability of this assembly and annotation provides a resource for further investigation of genomic organization in this species, and functional characterization of genes that may confer important phenotypic traits