Evaluation of the Regenerative Potential of Platelet-Lysate and Platelet-Poor Plasma Derived from the Cord Blood Units in Corneal Wound Healing Applications: An In Vitro Comparative Study on Corneal Epithelial Cells.

BACKGROUND: Cord blood platelet lysate (CB-PL) and cord blood platelet poor plasma (CB-PPP) have been applied with success in wound healing applications. Pathologies such as Sjogrens's Syndrome (SS) and chronic graft versus host disease (cGVHD) can lead to severe ophthalmology issues. The application of CB-PL and CB-PPP may be strongly considered for damaged cornea healing. This study aimed to the evaluation of the beneficial properties of CB-PL and CB-PPP in corneal wound healing applications. METHODS: Initially, the CB-PL and CB-PPP were produced from donated cord blood units (CBUs), followed by biochemical analysis. Corneal epithelial cells (CECs) were isolated from wistar rats and then cultured with medium containing 20% v/v either of CB-PL or CB-PPP. To define the impact of CB-PL and CB-PPP, biochemical, morphological analysis, scratch-wound assays, and immunoassays in CECs were performed. RESULTS: CB-PL and CB-PPP were characterized by good biochemical parameters, regarding their quality characteristics and biomolecule content. CECs' morphological features did not change after their cultivation with CB-PL or CB-PPP. A scratch wound assay and molecular analysis of CECs expanded with CB-PL indicated higher migratory capacity compared to those cultured with CB-PPP. CONCLUSION: CB-PL and CB-PPP exhibited good properties with respect to cell migration and proliferation, and could be considered an alternative source for eye drop production, to possibly be used in cornea wound healing applications.

Investigating the production of platelet lysate obtained from low volume Cord Blood Units: Focus on growth factor content and regenerative potential.

BACKGROUND: The regenerative potential of platelet lysate (PL) and platelet gel (PG) is mediated by the release of platelets (PLTs) growth factors. The aim of this study was the evaluation of the PL production utilizing low volume single Cord Blood Units (CBUs) and the comparison of the biomolecule content between PLs obtained from intermediate and high volume CBUs. METHODS: CBUs (n = 90) with volumes greater than 50 ml and initial platelet count > 150 × 109/L were used. CBUs were classified into the following groups: group A (50-80 ml), group B (81-110 ml) and group C (111-150 ml). The CBUs were centrifuged twice for the production of the platelet concentrate (PC), which was stored at - 80 °C for at least 48 h. Then, rapidly thawed and the biomolecule content was determined using commercial ELISA kits. The regenerative potential of PLs was evaluated using the scratch wound and in vitro angiogenesis assay. RESULTS: CBPL was produced from low volume single CBUs and contained 3.4 ± 0.3 ×109 PLTs. PL obtained from intermediate and high volume CBUs consisted of 10.2 ±â€¯0.3 and 16.1 ±â€¯0.4 × 109 PLTs. All PL groups were characterized by high biomolecule content. Gap closure was observed within 72 h after the wound assay initiation and the capillary tubes were formed in all study groups. CONCLUSION: This study provided significant evidence regarding the utilization of the low volume CBUs for the production of CB derivatives, thus can serve as healing mediators in regenerative medicine approaches.

Vineyard-mediated factors are still operative in spontaneous and commercial fermentations shaping the vinification microbial community and affecting the antioxidant and anticancer properties of wines.

The grapevine and vinification microbiota have a strong influence on the characteristics of the produced wine. Currently we have a good understanding of the role of vineyard-associated factors, like cultivar, vintage and terroir in shaping the grapevine microbiota. Notwithstanding, their endurance along the vinification process remains unknown. Thus, the main objective of our study was to determine how these factors influence (a) microbial succession during fermentation (i.e., bacterial and fungal) and (b) the antioxidant, antimutagenic and anticancer potential of the produced wines. These were evaluated under different vinification strategies (i.e., spontaneous V1, spontaneous with preservatives V2, commercial V3), employed at near full-scale level by local wineries, for two cultivars (Roditis and Sideritis), two terroir types, and two vintages. Cultivar and vintage were strong and persistent determinants of the vinification microbiota, unlike terroir whose effect became weaker from the vineyard, and early fermentation stages, where non-Saccharomyces yeasts, filamentous fungi (i.e., Aureobasidium, Cladosporium, Lachancea, Alternaria, Aspergillus, Torulaspora) and acetic acid bacteria (AAB) (Gluconobacter, Acetobacter, Komagataeibacter) dominated, to late fermentation stages where Saccharomyces and Oenococcus become prevalent. Besides vineyard-mediated factors, the vinification process employed was the strongest determinant of the fungal community compared to the bacterial community were effects varied per cultivar. Vintage and vinification type were the strongest determinants of the antioxidant, antimutagenic and anticancer potential of the produced wines. Further analysis identified significant positive correlations between members of the vinification microbiota like the yeasts Torulaspora debrueckii and Lachancea quebecensis with the anticancer and the antioxidant properties of wines in both cultivars. These findings could be exploited towards a microbiota-modulated vinification process to produce high-quality wines with desirable properties and enhanced regional identity.

DNA barcoding reveals different cestode helminth species in northern European marine and freshwater ringed seals.

Three subspecies of the ringed seal (Pusa hispida) are found in northeastern Europe: P. h. botnica in the Baltic Sea, P. h saimensis in Lake Saimaa in Finland, and P. h. ladogensis in Lake Ladoga in Russia. We investigated the poorly-known cestode helminth communities of these closely related but ecologically divergent subspecies using COI barcode data. Our results show that, while cestodes from the Baltic Sea represent Schistocephalus solidus, all worms from the two lakes are identified as Ligula intestinalis, a species that has previously not been reported from seals. The observed shift in cestode communities appears to be driven by differential availability of intermediate fish host species in marine vs. freshwater environments. Both observed cestode species normally infect fish-eating birds, so further work is required to elucidate the health and conservation implications of cestode infections in European ringed seals, whether L. intestinalis occurs also in marine ringed seals, and whether the species is able to reproduce in seal hosts. In addition, a deep barcode divergence found within S. solidus suggests the presence of cryptic diversity under this species name.

Grapevine wood microbiome analysis identifies key fungal pathogens and potential interactions with the bacterial community implicated in grapevine trunk disease appearance.

BACKGROUND: Grapevine trunk diseases (GTDs) is a disease complex caused by wood pathogenic fungi belonging to genera like Phaeomoniella, Phaeoacremonium, Fomitiporia, Eutypa and members of the family Botryosphaeriaceae. However, the co-occurrence of these fungi in symptomatic and asymptomatic vines at equivalent abundances has questioned their role in GTDs. Hence, we still lack a good understanding of the fungi involved in GTDs, their interactions and the factors controlling their assemblage in vines. We determined the fungal and bacterial microbiome in wood tissues of asymptomatic and symptomatic vines of three main Greek cultivars (Agiorgitiko, Xinomavro, Vidiano), each cultivated in geographically distinct viticultural zones, using amplicon sequencing. RESULTS: We noted that cultivar/biogeography (lumped factor) was the strongest determinant of the wood fungal microbiome (p < 0.001, 22.7%), while GTD symptoms condition had a weaker but still significant effect (p < 0.001, 3.5%), being prominent only in the cultivar Xinomavro. Several fungal Amplicon Sequence Variants (ASVs), reported as GTD-associated pathogens like Kalmusia variispora, Fomitiporia spp., and Phaemoniella chlamydosporα (most dominant in our study), were positively correlated with symptomatic vines in a cultivar/viticultural zone dependent manner. Random Forest analysis pointed to P. chlamydosporα, K. variispora, A. alternata and Cladosporium sp., as highly accurate predictors of symptomatic vines (0% error rate). The wood bacterial microbiome showed similar patterns, with biogeography/cultivar being the main determinant (p < 0.001, 25.5%) of its composition, followed by the GTD status of vines (p < 0.001, 5.2%). Differential abundance analysis revealed a universal positive correlation (p < 0.001) of Bacillus and Streptomyces ASVs with asymptomatic vines. Network analysis identified a significant negative co-occurrence network between these bacterial genera and Phaemoniella, Phaeoacrominum and Seimatosporium. These results point to a plant beneficial interaction between Bacillus/Streptomyces and GTD pathogens. CONCLUSIONS: Our study (a) provides evidence that GTD symptomatic plants support a wood fungal microbiome, showing cultivar and biogeography-dependent patterns, that could be used as a proxy to distinguish between healthy and diseased vines, (b) points to strong interactions between the bacterial and fungal wood microbiome in asymptomatic vines that should be further pursued in the quest for discovery of novel biocontrol agents.

Functionally Equivalent Variants in a Non-standard Variety and Their Implications for Universal Grammar: A Spontaneous Speech Corpus.

Findings from the field of experimental linguistics have shown that a native speaker may judge a variant that is part of her grammar as unacceptable, but still use it productively in spontaneous speech. The process of eliciting acceptability judgments from speakers of non-standard languages is sometimes clouded by factors akin to prescriptive notions of grammatical correctness. It has been argued that standardization enhances the ability to make clear-cut judgments, while non-standardization may result to grammatical hybridity, often manifested in the form of functionally equivalent variants in the repertoire of a single speaker. Recognizing the importance of working with corpora of spontaneous speech, this work investigates patterns of variation in the spontaneous production of five neurotypical, adult speakers of a non-standard variety in terms of three variants, each targeting one level of linguistic analysis: syntax, morphology, and phonology. The results reveal the existence of functionally equivalent variants across speakers and levels of analysis. We first discuss these findings in relation to the notions of competing, mixed, and fused grammars, and then we flesh out the implications that different values of the same variant carry for parametric approaches to Universal Grammar. We observe that intraspeaker realizations of different values of the same variant within the same syntactic environment are incompatible with the 'triggering-a-single-value' approach of parametric models, but we argue that they are compatible with the concept of Universal Grammar itself. Since the analysis of these variants is ultimately a way of investigating the status of Universal Grammar primitives, we conclude that claims about the alleged unfalsifiability of (the contents of) Universal Grammar are unfounded.

The Influence of Bilectalism and Non-standardization on the Perception of Native Grammatical Variants.

Research in speakers of closely related varieties has shown that bilectalism and non-standardization affect speakers' perception of the variants that exist in their native languages in a way that is absent from the performance of their monolingual peers. One possible explanation for this difference is that non-standardization blurs the boundaries of grammatical variants and increases grammatical fluidity. Affected by such factors, bilectals become less accurate in identifying the variety to which a grammatical variant pertains. Another explanation is that their differential performance derives from the fact that they are competent in two varieties. Under this scenario, the difference is due to the existence of two linguistic systems in the course of development, and not to how close or standardized these systems are. This study employs a novel variety-judgment task in order to elucidate which of the two explanations holds. Having administered the task to monolinguals, bilectals, and bilinguals, including heritage language learners and L1 attriters, we obtained a dataset of 16,245 sentences. The analysis shows differential performance between bilectal and bilingual speakers, granting support for the first explanation. We discuss the role of factors such as non-standardization and linguistic proximity in language development and flesh out the implications of the results in relation to different developmental trajectories.

Stochastic Optimization for an Analytical Model of Saltwater Intrusion in Coastal Aquifers.

The present study implements a stochastic optimization technique to optimally manage freshwater pumping from coastal aquifers. Our simulations utilize the well-known sharp interface model for saltwater intrusion in coastal aquifers together with its known analytical solution. The objective is to maximize the total volume of freshwater pumped by the wells from the aquifer while, at the same time, protecting the aquifer from saltwater intrusion. In the direction of dealing with this problem in real time, the ALOPEX stochastic optimization method is used, to optimize the pumping rates of the wells, coupled with a penalty-based strategy that keeps the saltwater front at a safe distance from the wells. Several numerical optimization results, that simulate a known real aquifer case, are presented. The results explore the computational performance of the chosen stochastic optimization method as well as its abilities to manage freshwater pumping in real aquifer environments.