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.     

Extension of the Terpene Chemical Space: the Very First Biosynthetic Steps

The structural diversity of terpenes is particularly notable and many studies are carried out to increase it further. In the terpene biosynthetic pathway this diversity is accessible from only two common precursors, i. e. isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). Methods recently developed (e. g. the Terpene Mini Path) have allowed DMAPP and IPP to be obtained from a two-step enzymatic conversion of industrially available isopentenol (IOH) and dimethylallyl alcohol (DMAOH) into their corresponding diphosphates. Easily available IOH and DMAOH analogues then offer quick access to modified terpenoids thus avoiding the tedious chemical synthesis of unnatural diphosphates. The aim of this minireview is to cover the literature devoted to the use of these analogues for widening the accessible terpene chemical space.     

Assessment of TSPAN Expression Profile and Their Role in the VSCC Prognosis

The role and prognostic value of tetraspanins (TSPANs) in vulvar squamous cell carcinoma (VSCC) remain poorly understood. We sought to primarily determine, at both the molecular and tissue level, the expression profile of the TSPANs CD9, CD63, CD81, and CD82 in archived VSCC samples (n = 117) and further investigate their clinical relevance as prognostic markers. Our studies led us to identify CD63 as the most highly expressed TSPAN, at the gene and protein levels. Multicomparison studies also revealed that the expression of CD9 was associated with tumor size, whereas CD63 upregulation was associated with histological diagnosis and vascular invasion. Moreover, low expression of CD81 and CD82 was associated with worse prognosis. To determine the role of TSPANs in VSCC at the cellular level, we assessed the mRNA levels of CD63 and CD82 in established metastatic (SW962) and non-metastatic (SW954) VSCC human cell lines. CD82 was found to be downregulated in SW962 cells, thus supporting its metastasis suppressor role. However, CD63 was significantly upregulated in both cell lines. Silencing of CD63 by siRNA led to a significant decrease in proliferation of both SW954 and SW962. Furthermore, in SW962 particularly, CD63-siRNA also remarkably inhibited cell migration. Altogether, our data suggest that the differential expression of TSPANs represents an important feature for prognosis of VSCC patients and indicates that CD63 and CD82 are likely potential therapeutic targets in VSCC.     

Effect of anionic substitution on the high temperature stability of polymer-derived SiOC glasses

In this study the high temperature stability (crystallization and decomposition) of two silicon oxycarbide glasses with a similar amount of free carbon (8.3 vs 9.6 wt%) but different content of Si-C bonds (SiC_0.22O_1.57 vs SiC_0.07O_1.86) is presented. The two SiOC glasses are obtained from the same precursor (2 µm methyl-silsesquioxane spheres) via pyrolysis at 1100°C in inert (Ar) or reactive (CO₂) atmospheres. Further annealing in Ar flow at temperatures above 1100°C and up to 1500°C is performed and the samples are characterized by Fourier Transformed Infrared Spectroscopy (FT-IR) and X-ray diffraction (XRD). For comparison purposes the same precursor was annealed in air flow to obtain SiO₂ and its high temperature evolution is also studied. Results suggest that the onset for the carbothermal reduction is not dependent on the amount of Si-C bonds. Moreover, contrary to what is usually reported in the scientific literature, silica phase present in the SiOC glasses does not show, in the same experimental conditions, superior crystallization resistance compared to pure silica glass.     

Therapeutic miR-21 Silencing Reduces Cardiac Fibrosis and Modulates Inflammatory Response in Chronic Chagas Disease

Chagas disease, caused by the parasite Trypanosoma cruzi (T. cruzi), remains a serious public health problem for which there is no effective treatment in the chronic stage. Intense cardiac fibrosis and inflammation are hallmarks of chronic Chagas disease cardiomyopathy (CCC). Previously, we identified upregulation of circulating and cardiac miR-21, a pro-fibrotic microRNA (miRNA), in subjects with CCC. Here, we explored the potential role of miR-21 as a therapeutic target in a model of chronic Chagas disease. PCR array-based 88 microRNA screening was performed in heart samples obtained from C57Bl/6 mice chronically infected with T. cruzi and serum samples collected from CCC patients. MiR-21 was found upregulated in both human and mouse samples, which was corroborated by an in silico analysis of miRNA-mRNA target prediction. In vitro miR-21 functional assays (gain-and loss-of-function) were performed in cardiac fibroblasts, showing upregulation of miR-21 and collagen expression upon transforming growth factor beta 1 (TGFβ1) and T. cruzi stimulation, while miR-21 blockage reduced collagen expression. Finally, treatment of T. cruzi-infected mice with locked nucleic acid (LNA)-anti-miR-21 inhibitor promoted a significant reduction in cardiac fibrosis. Our data suggest that miR-21 is a mediator involved in the pathogenesis of cardiac fibrosis and indicates the pharmacological silencing of miR-21 as a potential therapeutic approach for CCC.     

Optimization of a Fabric Softener Formulation with an Electrochemical Sensor and Streaming Potential Measurements

An electrochemical, cationic, surfactant-selective sensor based on multiwalled carbon nanotubes (MWCNT) functionalized with a sulfate group and the cetylpyridinium ion (MWCNT–OSO₃⁻CP⁺) as a sensing material was used for optimization of the formulation of a fabric softener. Potentiometric titrations were performed and response measurements were obtained using four cationic surfactants (CS) of technical grade and four CS of analytical grade. The slope closest to Nernstian was obtained for di-(tallow carboxyethyl) hydroxyethyl methylammonium methosulfate (MAS) (59.5 ± 1.1 mV/decade of activity in water and 57.5 ± 1.3 mV/decade of activity in CaCl₂). When using CS as analytes in potentiometric titrations, the best accuracy (99.8%) was obtained when using MAS; therefore, it was chosen as the CS for the fabric softener formulation. Due to the better properties of fabric softeners with silicone in their formulations, four silicones at several concentration levels were used as potential additives. Based on the stability and viscosity of the system, the diquaternary polydimethylsiloxane (DPS) (w = 0.19%) was chosen for the fabric softener formulation. The pH did not significantly influence the potential when in the range of 3–8 or the recovery of potentiometric titrations when in the range of 3–7. The application profile of the CS was assessed through streaming potential measurements of reference fabrics in an electrokinetic analyzer. The obtained electrokinetic parameters indicated on lag in adsorption of model fabric softener (MFS) based on MAS (w = 9%), with the addition of silicone DPS (MFS 3), on cotton and polyester fabrics, but advantage in stability when compared with other MFS investigated.     

Coffee with jelly or unbuttered toast: Commissions and omissions are dissociable aspects of everyday action impairment in Alzheimer's disease.

Relative to our understanding of the memory and language deficits associated with Alzheimer's disease (AD), little is known about problems with everyday action performance (i.e., meal preparation, grooming). The resource theory proposes that everyday action problems are best explained by a unitary deficit in general cognitive resources. However, recent research suggests that omission and commission errors may reflect dissociable aspects of action impairment, with only omissions associated with resource limitations. This study examined everyday action performance in 70 participants with AD who also underwent a neuropsychological evaluation. First, correlation and principal component analyses were performed to examine the construct(s) that might explain everyday action impairment. Second, relations between everyday task component(s) and neuropsychological tests were examined by using correlation and regression analyses. Third, differences in everyday action error patterns were examined among participants of comparable overall impairment levels. Results showed omission and commission errors were uncorrelated and distinct components of everyday action performance, predicted by different neuropsychological tests, and differentially distributed even among participants with comparable overall impairment. (PsycINFO Database Record (c) 2010 APA, all rights reserved)