Qualitative and Quantitative Comparison of Plasma Exosomes from Neonates and Adults

Exosomes are extracellular vesicles that contain nucleic acids, lipids and metabolites, and play a critical role in health and disease as mediators of intercellular communication. The majority of extracellular vesicles in the blood are platelet-derived. Compared to adults, neonatal platelets are hyporeactive and show impaired granule release, associated with defects in Soluble N-ethylmaleimide-sensitive fusion Attachment protein REceptor (SNARE) proteins. Since these proteins participate in biogenesis of exosomes, we investigated the potential differences between newborn and adult plasma-derived exosomes. Plasma-derived exosomes were isolated by ultracentrifugation of umbilical cord blood from full-term neonates or peripheral blood from adults. Exosome characterization included size determination by transmission electron microscopy and quantitative proteomic analysis. Plasma-derived exosomes from neonates were significantly smaller and contained 65% less protein than those from adults. Remarkably, 131 proteins were found to be differentially expressed, 83 overexpressed and 48 underexpressed in neonatal (vs. adult) exosomes. Whereas the upregulated proteins in plasma exosomes from neonates are associated with platelet activation, coagulation and granule secretion, most of the underexpressed proteins are immunoglobulins. This is the first study showing that exosome size and content change with age. Our findings may contribute to elucidating the potential “developmental hemostatic mismatch risk” associated with transfusions containing plasma exosomes from adults.     

P16 and HPV Genotype Significance in HPV-Associated Cervical Cancer—A Large Cohort of Two Tertiary Referral Centers

The expression of p16 is a good surrogate of human papillomavirus (HPV) infection in HPV-associated cancers. The significance of p16 expression, HPV genotype and genera in the outcome of patients with HPV-associated cervical cancer (CC) is unclear. Our aim is to ascertain the prognostic significance of these factors. Data from 348 patients (median age: 47.5 years old) with CC, diagnosed in two referral centers, were retrospectively collected. Advanced disease (FIGO2018 IB2-IV) was present in 68% of patients. A single HPV genotype was identified in 82.8% of patients. The most common HPVs were HPV16 (69%) and HPV18 (14%). HPV genera reflected this distribution. HPV16 tumors presented at an earlier stage. P16 was negative in 18 cases (5.2%), 83.3% of which were squamous cell carcinomas. These cases occurred in older patients who tended to have advanced disease. In the univariate analysis, HPV16 (HR: 0.58; p = 0.0198), α-9 genera (HR: 0.37; p = 0.0106) and p16 overexpression (HR: 0.54; p = 0.032) were associated with better survival. HPV16 (HR: 0.63; p = 0.0174) and α-9 genera (HR: 0.57; p = 0.0286) were associated with less relapse. In the multivariate analysis, only the International Federation of Gynecology and Obstetrics (FIGO) stage retained an independent prognostic value. HPV16, α-9 genera and p16 overexpression were associated with better survival, although not as independent prognostic factors. Patients with p16-negative HPV-associated CC were older, presented with advanced disease and had worse prognosis.     

Validation of a Gas Chromatography-Mass Spectrometry Method for the Measurement of the Redox State Metabolic Ratios Lactate/Pyruvate and β-Hydroxybutyrate/Acetoacetate in Biological Samples

The metabolic ratios lactate/pyruvate and β-hydroxybutyrate/acetoacetate are considered valuable tools to evaluate the in vivo redox cellular state by estimating the free NAD+/NADH in cytoplasm and mitochondria, respectively. The aim of the current study was to validate a gas-chromatography mass spectrometry method for simultaneous determination of the four metabolites in plasma and liver tissue. The procedure included an o-phenylenediamine microwave-assisted derivatization, followed by liquid-liquid extraction with ethyl acetate and silylation with bis(trimethylsilyl)trifluoroacetamide:trimethylchlorosilane 99:1. The calibration curves presented acceptable linearity, with a limit of quantification of 0.001 mM for pyruvate, β-hydroxybutyrate and acetoacetate and of 0.01 mM for lactate. The intra-day and inter-day accuracy and precision were within the European Medicines Agency’s Guideline specifications. No significant differences were observed in the slope coefficient of three-point standard metabolite-spiked curves in plasma or liver and water, and acceptable recoveries were obtained in the metabolite-spiked samples. Applicability of the method was tested in precision-cut liver rat slices and also in HepG2 cells incubated under different experimental conditions challenging the redox state. In conclusion, the validated method presented good sensitivity, specificity and reproducibility in the quantification of lactate/pyruvate and β-hydroxybutyrate/acetate metabolites and may be useful in the evaluation of in vivo redox states.     

Land Use Effect on the CN Model Parameters in a Tropical Dry Environment

Dry tropical forests account for over 1,000,000 km², and there is still lack of knowledge on their hydrologic processes. The curve number (CN) hydrologic model developed by the Natural Resources Conservation Service (NRCS) is widely applied for runoff determination in various parts of the world, but not so in tropical semiarid regions. This study analyzes the impact of land use changes on the CN model in a tropical semiarid environment, in two catchments of native dry tropical forest and thinned dry tropical forest land use from 2009 to 2012. The CN model was calibrated and validated for the NRCS recommended initial abstraction ratio λ = 0.2, and for λ evaluated from rainfall and runoff data. A reliability analysis was performed using Monte Carlo simulation. Model goodness-of-fit was assessed with statistical criteria. A total of 42 and 40 rainfall-runoff events were analyzed for the native and thinned dry tropical forest, respectively. Characteristic λ values of 0.15 and 0.11 were determined for the two respective catchments. Although CN values were similar for both land uses, CN_λ=0.20 = 80 and CN_{median λ} = 77, the thinned catchment showed a higher CN model parameters variability. The CN model was more sensitive to variations of CN values than to those of λ. This study showed that no matter the vegetation management in a dry tropical forest environment, modeled runoff is not affected by λ, but rather affected by CN, which represents soil, landuse and management.     

Polymerization of diisopropyl fumarate under microwave irradiation

Diisopropyl fumarate (DiPF), a representative monomer of dialkyl fumarates, was polymerized by microwave irradiation at three different powers (140, 210, and 280 W), using a domestic microwave oven. The nature and concentration of initiators [2,2′‐azobisisobutyronitrile (AIBN) and benzoyl peroxide (BP)], power and energy of microwave irradiation on the conversion, weight average molecular weight (M_w), and polydispersity index (M_w/M_n) were analyzed. The results indicate that the microwave conditions have a significant nonthermal effect in enhancing the polymerization rate of DiPF. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 103: 3785–3791, 2007     

Effect of catalysts in the quality of syngas and by-products obtained by co-gasification of coal and wastes. 1. Tars and nitrogen compounds abatement

The aim of this work is to analyse the possibility of using co-gasification technology to process coal mixed with wastes to take profit of its energy content and at the same time to minimize the environmental impact associated with the use of wastes and to diminish the costs of flue gas treatment. The addition to coal of different types of materials, like: pine based waste, petcoke and polyethylene (PE), was not found to give rise to any operational problems, regarding both the feeding system and gasification process and led to higher energy conversions, however, the gas presented higher tars and hydrocarbons content. Several catalysts were tested, such as, dolomite, olivine, nickel and magnesium oxides, zinc oxides and cobalt and molybdenum oxides. Catalyst action was analysed in tars release and also in ammonia compounds reduction. The presence of catalysts allowed increasing hydrogen release, whilst there was a decrease in hydrocarbons and tars contents. A nickel-magnesium oxide was the catalyst that led to the highest reduction in hydrocarbons and tars. This catalyst also led to the lowest NH₃ content in the fuel gas produced, due to the catalyst efficiency in NH₃ destruction.     

Evaluation of the thermal and mechanical properties of poly(ε‐caprolactone), low‐density polyethylene,and their blends

The thermal and mechanical properties of low‐density polyethylene (LDPE), poly(ε‐caprolactone) (PCL), and their blends were evaluated. Differential scanning calorimetry showed that increasing the PCL content of the blend did not change the LDPE melting temperature, but reduced the crystallinity by up to 16.8%. This behavior was related to interactions between the PCL chains and the crystalline phase of LDPE. Tensile strength and elongation at break values for the blends were lower than those for the pure polymers, which suggested an incompatibility between the polymers. The values for Young's modulus under tensile increased when PCL was added to LDPE. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 91:3909–3914, 2004     

Structural characterization of reactor blends of polypropylene and ethylene‐propylene rubber

Blends of isotactic polypropylene (PP), ethylene‐propylene rubber copolymer (EPR), and ethylene‐propylene crystalline copolymer (EPC) can be produced through in situ polymerization processes directly in the reactor and blends with different structure and composition can be obtained. In this work we studied the structure of five reactor‐made blends of PP, EPR, and EPC produced by a Ziegler‐Natta catalyst system. The composition of EPR was related to the ratio between ethylene and propylene used in the copolymerization step. The ethylene content in the EPR was in the range of 50–70 mol %. The crystallization behavior of PP and EPC in the blends was influenced by the presence of the rubber, and some specific interactions between the components could be established. By preparative temperature rising elution fractionation (P‐TREF) analysis, the isolation and characterization of crystalline EPC fractions were made. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 92: 2155–2162, 2004     

Effect of low‐temperature plasma and chitosan treatment on wool dyeing with Acid Red 27

This study examines in detail the influence of low‐temperature plasma and biopolymer chitosan treatments on wool dyeability. Wool knitted fabrics were treated and characterized by whiteness and shrink‐resistance measurements. Surface modification was assessed by contact‐angle measurements of human hair fibers, which were used as a model to study the wetting properties of the treated wool knitted fabrics. The dyeing behavior was assessed from the diffusion mechanism point of view. The dyeing kinetics were measured at two different pHs (4.2 and 6.5) and three different temperatures (60, 85, and 100°C) to gain information about the contribution of the surface modification treatment to the dyeing mechanism. The exhaustion and reflectance data were compared, and the apparent diffusion coefficients were calculated. On the basis of the obtained results, a model for the dyeing mechanism of the chitosan treated wool was proposed. When treated with chitosan, the polymer sheath spread on the surface of the fibers acted as a predominant dyeing site in very short dyeing times, thus interacting with the dye and in later stages imparting the dye to the wool fiber. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 97: 2204–2214, 2005     

A description of phase inversion behaviour in agitated liquid-liquid dispersions under the influence of the Marangoni effect

The influence of the Marangoni effect on phase inversion behaviour is examined by integrating a microscopic study of the drop coalescence process, in which thin film drainage in the presence of insoluble surfactant occurs, into a macroscopic phase inversion model which has been developed previously using a Monte Carlo technique. This is achieved via an immobility factor, obtained from a comparison of the film drainage times for surfactant-laden systems and surfactant-free systems as a function of the drop approach velocity, surface Péclet number, initial surfactant concentration and the Hamaker constant, which is then used to modify the coalescence probability in the phase inversion model. On the one hand, the results indicate that the Marangoni effect removes any influence that the viscosity ratio has on phase inversion due to immobilisation of the interface, thus shielding the flow in the film from the effects of the flow in the dispersed phase; the point at which phase inversion occurs therefore tends towards equivolume holdups with the addition of surfactant. On the other hand, when comparisons are made with pure systems in which surfactant is absent, the system is seen to be either stabilised or de-stabilised from inversion depending on the viscosity ratio of the system. This is attributed to the influence of surfactant on the dispersion morphologies on either side of the inversion (i.e. water-in-oil dispersions and oil-in-water dispersions) and depends on the dispersed phase holdup; the Marangoni effect is felt stronger when the dispersed phase holdup is low.