Comprehensive Profiling of Secretome Formulations from Fetal- and Perinatal Human Amniotic Fluid Stem Cells

We previously reported that c-KIT+ human amniotic-fluid derived stem cells obtained from leftover samples of routine II trimester prenatal diagnosis (fetal hAFS) are endowed with regenerative paracrine potential driving pro-survival, anti-fibrotic and proliferative effects. hAFS may also be isolated from III trimester clinical waste samples during scheduled C-sections (perinatal hAFS), thus offering a more easily accessible alternative when compared to fetal hAFS. Nonetheless, little is known about the paracrine profile of perinatal hAFS. Here we provide a detailed characterization of the hAFS total secretome (i.e., the entirety of soluble paracrine factors released by cells in the conditioned medium, hAFS-CM) and the extracellular vesicles (hAFS-EVs) within it, from II trimester fetal- versus III trimester perinatal cells. Fetal- and perinatal hAFS were characterized and subject to hypoxic preconditioning to enhance their paracrine potential. hAFS-CM and hAFS-EV formulations were analyzed for protein and chemokine/cytokine content, and the EV cargo was further investigated by RNA sequencing. The phenotype of fetal- and perinatal hAFS, along with their corresponding secretome formulations, overlapped; yet, fetal hAFS showed immature oxidative phosphorylation activity when compared to perinatal ones. The profiling of their paracrine cargo revealed some differences according to gestational stage and hypoxic preconditioning. Both cell sources provided formulations enriched with neurotrophic, immunomodulatory, anti-fibrotic and endothelial stimulating factors, and the immature fetal hAFS secretome was defined by a more pronounced pro-vasculogenic, regenerative, pro-resolving and anti-aging profile. Small RNA profiling showed microRNA enrichment in both fetal- and perinatal hAFS-EV cargo, with a stably- expressed pro-resolving core as a reference molecular signature. Here we confirm that hAFS represents an appealing source of regenerative paracrine factors; the selection of either fetal or perinatal hAFS secretome formulations for future paracrine therapy should be evaluated considering the specific clinical scenario.     

Probabilistic Quantitative Temporal Constraints: Representing,Reasoning, and Query Answering

In many applications and domains, temporal constraints between actions, and their probabilities play an important role. We propose the first approach in the literature coping with probabilistic quantitative constraints. To achieve such a challenging goal, we extend the widely used simple temporal problem (STP) framework to consider probabilities. Specifically, we propose i) a formal representation of probabilistic quantitative constraints, ii) an algorithm, based on the operations of intersection and composition, for the propagation of such temporal constraints, and iii) facilities to support query answering on a set of such constraints. As a result, we provide users with the first homogeneous method supporting the treatment (representing, reasoning, and querying) of probabilistic quantitative constraints, as required by many applications and domains.     

Effect of Chain-Length and Countersurface on the Tribochemistry of Bulk Zinc Polyphosphate Glasses

Zinc polyphosphate glasses are considered to be chiefly responsible for the anti-wear efficiency of ZnDTP tribofilms. In this work, the tribochemical properties of amorphous bulk zinc polyphosphates of different chain lengths (ranging from zinc metaphosphate to zinc pyrophosphate) have been investigated. Tribological tests on bulk polyphosphate discs have been carried out using steel and quartz balls as counter-surfaces in a poly-??-olefin (PAO) bath at room temperature. The composition in the wear track and on the contact region of the balls has been monitored by small-area and imaging X-ray photoelectron spectroscopy (i-XPS). The XPS analysis revealed that the composition of short-chain-length polyphosphates remained unchanged following tribological stress. Long-chain-length polyphosphates are depolymerized in the wear track as a consequence of a tribochemical reaction. By comparing the results obtained using quartz and steel balls, it could be observed that while the reaction of iron oxide with the polyphosphates certainly plays a role in the depolymerization of the samples under sliding conditions, pressure and shear stress alone and also in the presence of water or oil-oxidized species are able to depolymerize the glass when an inert material is used as counterpart; the composition of the wear track, in this case, is dependent on the applied load. All samples were able to form an adhesive, glassy transfer film on both steel and quartz balls, but the short-chain-length polyphosphates showed a lower friction coefficient and wear coefficient. The results suggest a third-body mechanism with the polyphosphates acting as a solid lubricant. Differences in tribological behavior of the different-chain-length polyphosphates are attributable to their mechanical and rheological properties.     

Evidence for electro-chemical interactions between multi-walled carbon nanotubes and human macrophages

Carbon multi-walled nanotubes (MWCNTs) may have several dangerous effects on different cell systems, but the mechanisms responsible for their cytotoxicity are not well known yet. At present, very little is known about the electrical interactions between nanomaterials and cells. We aimed to verify whether MWCNT electrical properties could affect the so called “charge-sensitive” cell parameters, interacting with cellular electrical activity. Human macrophages were challenged with two fully characterised MWCNT samples, one tested as-prepared (MWCNT), the other one purified (by annealing at 2400 °C) and better electro-conductive (a-MWCNT). Our findings show that a-MWCNTs are less cytotoxic but possess a higher inflammatory potential, as compared to MWCNTs. Moreover, only annealed and better conductive MWCNTs affect significantly the mitochondrial membrane polarity, the intracellular pH and the reorganisation of cytoskeleton actin filaments, cell functions strictly dependent on electro-chemical mechanisms. Based on our results, there is evidence for electro-chemical interactions taking place between cell membranes and electro-conductive MWCNTs. Such a specific behaviour could have wide-range applications in the biomedical field, not only concerning those cellular systems (neuronal and bone cells) sensitive to electrical stimuli, but also other cell systems.     

The content of proteic and nonproteic (free and protein-bound) tryptophan in quinoa and cereal flours

The content of proteic and nonproteic (free and protein-bound) tryptophan and of proteins in quinoa, wheat, rice, maize, barley, oat, rye, spelt, sorghum and millet flours was determined. Protein content and proteic tryptophan of quinoa were similar to that of wheat and spelt, but higher than in other cereals. Free tryptophan in quinoa flour showed values similar to those of wheat, oat and sorghum Kalblank, lower than those of barley, spelt and pearl millet, but higher than in rice, maize, rye, sorghum DK 34 – Alabama hybrid. In addition, nonproteic tryptophan appears bound both to water soluble proteins and to proteins soluble at pH 8.9. The results are discussed regarding the importance of the nonprotein tryptophan fraction, the only one able to enter the brain, that is more easily absorbed, so guarantees a greater amount available for uptake by the central nervous system.     

Volatile compounds and capsaicinoid content of fresh hot peppers (Capsicum annuum L.) of different Calabrian varieties

BACKGROUND: Because of the great variability in size, shape, colour, pungency and aroma of Calabrian Capsicum annuum(hot pepper), this study was carried out to furnish producers with information on volatile compounds and capsaicinoid content useful for the genetic improvement of this species. Fully ripened fresh peppers of the most widely cultivated varieties were studied. Capsaicinoid content was determined by reverse phase high‐performance liquid chromatography/diode array detection (RP‐HPLC/DAD). Volatile compounds were determined by headspace solid phase microextraction/gas chromatography/mass spectrometry (HS‐SPME/GC/MS). RESULTS: Sixty‐four volatile compounds were identified, the main classes being alcohols, aldehydes, terpenes and aliphatic branched‐chain hydrocarbons. The ‘Naso di cane’ (C. annuumvar. abbreviatum) variety contained the lowest amount of volatiles, especially alcohols and aldehydes, while the highest levels of capsaicin and dihydrocapsaicin were found in ‘Vulcan’ and ‘Corno di capra’ varieties. CONCLUSION: From statistical evaluation of the data obtained, each variety showed a typical composition of capsaicinoids and volatiles determining the taste and odour of the hot peppers. These results should be useful in the context of research aimed at selecting varieties with the best sensory characteristics. Copyright © 2009 Society of Chemical Industry     

Design of Experiments (DOE) for the Optimization of Titania–hydroxyapatite Functionally Graded Coatings

Titania–hydroxyapatite functionally graded coatings were deposited on titanium alloy substrates by plasma spraying. Because it was necessary to spray together the titania and the hydroxyapatite powders to obtain the graded system, the first target of the present study was to optimize the process parameters in order to obtain a high-quality coating. A 2³ Design of Experiments was applied to define the optimal values of plasma torch power, hydrogen flux, and spraying distance. This defined set of parameters (38 kW, 5 SLPM, and 90 mm, respectively) was used to spray the most promising graded coating, which was characterized and postheat treated.     

Identification of free and bound volatile compounds as typicalness and authenticity markers of non-aromatic grapes and wines through a combined use of mass spectrometric techniques

Molecular approaches by means of a combined use of mass spectrometric techniques can be important in order to open new possibilities in the differentiation and defense of typical products; in this study, a possible approach to the analysis of varietal volatile compounds and some precursors of a non-aromatic grape variety (Falanghina cv., Vitis vinifera L.) was traced through a combined use of techniques based on mass spectrometry (GC/MS, LC/ESI-MS, MALDI-TOF-MS). Dominant terpene compounds (limonene, cis-furanlinalool oxide, geraniol, 4-carene, myrcene, linalool, -terpineol), terpene-derivatives (bornyl acetate, menthol), terpene glycosides (glucosides, arabinosylglucosides and rhamnosylglucosides of linalool and geraniol), and norisoprenoids (β-damascenone) were identified in grapes and monovarietal wines, overcoming the analytical difficulties deriving from the low concentration of these compounds strictly related to the variety. The potential release of varietal volatile compounds from the grapes was also explored by enzyme hydrolysis.     

The Ketogenic Diet Reduces the Harmful Effects of Stress on Gut Mitochondrial Biogenesis in a Rat Model of Irritable Bowel Syndrome

Functional alterations in irritable bowel syndrome have been associated with defects in bioenergetics and the mitochondrial network. Effects of high fat, adequate-protein, low carbohydrate ketogenic diet (KD) involve oxidative stress, inflammation, mitochondrial function, and biogenesis. The aim was to evaluate the KD efficacy in reducing the effects of stress on gut mitochondria. Newborn Wistar rats were exposed to maternal deprivation to induce IBS in adulthood. Intestinal inflammation (COX-2 and TRL-4); cellular redox status (SOD 1, SOD 2, PrxIII, mtDNA oxidatively modified purines); mitochondrial biogenesis (PPAR-γ, PGC-1α, COX-4, mtDNA content); and autophagy (Beclin-1, LC3 II) were evaluated in the colon of exposed rats fed with KD (IBD-KD) or standard diet (IBS-Std), and in unexposed controls (Ctrl). IBS-Std rats showed dysfunctional mitochondrial biogenesis (PPAR-γ, PGC-1α, COX-4, and mtDNA contents lower than in Ctrl) associated with inflammation and increased oxidative stress (higher levels of COX-2 and TLR-4, SOD 1, SOD 2, PrxIII, and oxidatively modified purines than in Ctrl). Loss of autophagy efficacy appeared from reduced levels of Beclin-1 and LC3 II. Feeding of animals with KD elicited compensatory mechanisms able to reduce inflammation, oxidative stress, restore mitochondrial function, and baseline autophagy, possibly via the upregulation of the PPAR-γ/PGC-1α axis.