Development and Analysis of Three Diesel Particulate-Related Standard Reference Materials for the Determination of Chemical,Physical, and Biological Characteristics

The National Institute of Standards and Technology (NIST) has recently issued three Standard Reference Materials (SRMs) related to diesel particulate matter: SRM 1650a, Diesel Particulate Matter; SRM 2975, Diesel Particulate Matter (Industrial Forklift); and SRM 1975, Diesel Particulate Extract. These three materials have certified and reference concentrations for selected polycyclic aromatic hydrocarbons (PAHs) including many alkyl-substituted compounds. SRM 1650a is the reissue of an existing material and SRMs 2975 and 1975 represent new materials. The characterization of the PAH content of these three diesel particulate-related SRMs is presented with a discussion of the approach for the certification of PAH concentrations. In addition, the physical and biological characterization of these materials is discussed. A review of customer reported uses of these SRMs in the chemical, physical, and biological sciences is also provided.     

OPTIMAL KINETIC PARAMETERS AND BATCH MODELING FOR PECTIN HYDROLYSIS TO GALACTURONIC ACID WITH PECTINEX ULTRA SP-L ENZYME

Galacturonic acid is a monosaccharide obtained by pectin hydrolysis and a suitable substrate to produce bioethanol by fermentation. This article focuses on quantification of citrus pectin hydrolysis to galacturonic acid and provides new, reliable kinetic parameters for the Michaelis-Menten equation when the well-known commercial Pectinex Ultra SP-L is employed as enzyme. They are: r _max  = 1.10 g/(L min), K _m  = 10.42 g/L, and K _IGA  = 10.05 g/L, as obtained with a great accuracy by a nonlinear regression method and confirmed by the three classical linearization procedures (Lineweaver-Burk, Langmuir, and Eadie-Hofstee). The quantification of product inhibition has been achieved, with its inclusion in the rate equation. A batch reactor model yields perfect agreement between predictions and experiments, even under conditions different from those on which the parameters had been determined by regression.     

Mechanical analysis of structural adhesive for marine joints

A structural adhesive from I.C.R., X07 Extreme®, was fully characterised through tensile test, static and dynamic lap-shear strength test, cleavage peel test and DSC. Lap-shear tests were performed by using either aluminium, stainless steel and glass fibre reinforced polymer as substrate and after aging the specimen under extreme environmental conditions. The performance of this novel adhesive was compared to that of a structural adhesive of a competitor. The work summarises the typical procedures intended for the qualification of a structural adhesive for marine joints.     

Potential Mechanisms Linking Atherosclerosis and Increased Cardiovascular Risk in COPD: Focus On Sirtuins

The development of atherosclerosis is a multi-step process, at least in part controlled by the vascular endothelium function. Observations in humans and experimental models of atherosclerosis have identified monocyte recruitment as an early event in atherogenesis. Chronic inflammation is associated with ageing and its related diseases (e.g., atherosclerosis and chronic obstructive pulmonary disease). Recently it has been discovered that Sirtuins (NAD⁺-dependent deacetylases) represent a pivotal regulator of longevity and health. They appear to have a prominent role in vascular biology and regulate aspects of age-dependent atherosclerosis. Many studies demonstrate that SIRT1 exhibits anti-inflammatory properties in vitro (e.g., fatty acid-induced inflammation), in vivo (e.g., atherosclerosis, sustainment of normal immune function in knock-out mice) and in clinical studies (e.g., patients with chronic obstructive pulmonary disease). Because of a significant reduction of SIRT1 in rodent lungs exposed to cigarette smoke and in lungs of patients with chronic obstructive pulmonary disease (COPD), activation of SIRT1 may be a potential target for chronic obstructive pulmonary disease therapy. We review the inflammatory mechanisms involved in COPD-CVD coexistence and the potential role of SIRT1 in the regulation of these systems.     

Thermo-mechanical characterization of fumed silica-epoxy nanocomposites

DGEBA-based epoxy nanocomposites filled with various amounts of fumed silica nanopowders (10, 20 and 30 phr corresponding to 3.3, 6.4 and 9.2% by volume) were prepared by a solvent assisted dispersion procedure. The obtained nanocomposites were analyzed by means of differential scanning calorimetry, dynamic-mechanical thermal analysis, uniaxial tensile tests on un-notched samples and three-point bending tests on notched samples.

All the thermal and mechanical properties showed non-monotonic trends with relative minima as the silica content increased. A trend inversion in the physical properties was detected at the highest filler level tested, which was not previously observed for epoxy systems filled with comparable amounts of unmodified silica. The behavior could be explained by considering a reduction of cross-linking degree of the epoxy matrix due to the huge viscosity increase induced by the silica nanoparticles during composites preparation. On the other hand, the physical immobilization of the cross-linked matrix can be supposed to be responsible for the inversion of the properties trend at the highest filler level, which is presumably very close to the percolation threshold.     

Acute β-Hydroxy-β-Methyl Butyrate Suppresses Regulators of Mitochondrial Biogenesis and Lipid Oxidation While Increasing Lipid Content in Myotubes

Leucine modulates synthetic and degradative pathways in muscle, possibly providing metabolic benefits for both athletes and diseased populations. Leucine has become popular among athletes for improving performance and body composition, however little is known about the metabolic effects of the commonly consumed leucine‐derived metabolite β‐hydroxy‐β‐methyl butyrate (HMB). Our work measured the effects of HMB on metabolic protein expression, mitochondrial content and metabolism, as well as lipid content in skeletal muscle cells. Specifically, cultured C2C12 myotubes were treated with either a control or HMB ranging from 6.25 to 25 μM for 24 h and mRNA and/or protein expression, oxygen consumption, glucose uptake, and lipid content were measured. Contrary to leucine's stimulatory effect on metabolism, HMB‐treated cells exhibited significantly reduced regulators of lipid oxidation including peroxisome proliferator‐activated receptor alpha (PPARα) and PPARβ/δ, as well as downstream target carnitine palmitoyl transferase, without alterations in glucose or palmitate oxidation. Furthermore, HMB significantly inhibited activation of the master regulator of energetics, AMP‐activated protein kinase. As a result, HMB‐treated cells also displayed reduced total mitochondrial content compared with true control or cells equivocally treated with leucine. Additionally, HMB treatment amplified markers of lipid biosynthesis (PPARγ and fatty acid synthase) as well as consistently promoted elevated total lipid content versus control cells. Collectively, our results demonstrate that HMB did not improve mitochondrial metabolism or content, and may promote elevated cellular lipid content possibly through heightened PPARγ expression. These observations suggest that HMB may be most beneficial for populations interested in stimulating anabolic cellular processes.     

The L467F-F508del Complex Allele Hampers Pharmacological Rescue of Mutant CFTR by Elexacaftor/Tezacaftor/Ivacaftor in Cystic Fibrosis Patients: The Value of the Ex Vivo Nasal Epithelial Model to Address Non-Responders to CFTR-Modulating Drugs

Loss-of-function mutations of the CFTR gene cause cystic fibrosis (CF) through a variety of molecular mechanisms involving altered expression, trafficking, and/or activity of the CFTR chloride channel. The most frequent mutation among CF patients, F508del, causes multiple defects that can be, however, overcome by a combination of three pharmacological agents that improve CFTR channel trafficking and gating, namely, elexacaftor, tezacaftor, and ivacaftor. This study was prompted by the evidence of two CF patients, compound heterozygous for F508del and a minimal function variant, who failed to obtain any beneficial effects following treatment with the triple drug combination. Functional studies on nasal epithelia generated in vitro from these patients confirmed the lack of response to pharmacological treatment. Molecular characterization highlighted the presence of an additional amino acid substitution, L467F, in cis with the F508del variant, demonstrating that both patients were carriers of a complex allele. Functional and biochemical assays in heterologous expression systems demonstrated that the double mutant L467F-F508del has a severely reduced activity, with negligible rescue by CFTR modulators. While further studies are needed to investigate the actual prevalence of the L467F-F508del allele, our results suggest that this complex allele should be taken into consideration as plausible cause in CF patients not responding to CFTR modulators.     

The Impact of Nonalcoholic Fatty Liver Disease on Renal Function in Children with Overweight/Obesity

The association between nonalcoholic fatty liver disease (NAFLD) and chronic kidney disease has attracted interest and attention over recent years. However, no data are available in children. We determined whether children with NAFLD show signs of renal functional alterations, as determined by estimated glomerular filtration rate (eGFR) and urinary albumin excretion. We studied 596 children with overweight/obesity, 268 with NAFLD (hepatic fat fraction ≥5% on magnetic resonance imaging) and 328 without NAFLD, and 130 healthy normal-weight controls. Decreased GFR was defined as eGFR < 90 mL/min/1.73 m². Abnormal albuminuria was defined as urinary excretion of ≥30 mg/24 h of albumin. A greater prevalence of eGFR < 90 mL/min/1.73 m² was observed in patients with NAFLD compared to those without liver involvement and healthy subjects (17.5% vs. 6.7% vs. 0.77%; p < 0.0001). The proportion of children with abnormal albuminuria was also higher in the NAFLD group compared to those without NAFLD, and controls (9.3% vs. 4.0% vs. 0; p < 0.0001). Multivariate logistic regression analysis revealed that NAFLD was associated with decreased eGFR and/or microalbuminuria (odds ratio, 2.54 (confidence interval, 1.16–5.57); p < 0.05) independently of anthropometric and clinical variables. Children with NAFLD are at risk for early renal dysfunction. Recognition of this abnormality in the young may help to prevent the ongoing development of the disease.     

Visible spectroelectrochemical characterization of Geobacter sulfurreducens biofilms on optically transparent indium tin oxide electrode

We report visible spectroelectrochemical (SEC) characterization of cytochrome c₅₅₂ (cyt c₅₅₂) in viable Geobacter sulfurreducens biofilms on tin-doped indium oxide (ITO) electrodes poised at 0.24 V vs. SHE. G. sulfurreducens biofilms were grown in minimal medium with acetate as electron donor (turnover conditions), followed by 24 h incubation in electron donor-depleted medium (non-turnover conditions). The electronic absorption spectra of G. sulfurreducens biofilms showed the lowest energy absorption band in the reduced state at 552 nm, which indicated excess of cyt c₅₅₂ in the biofilm. The spectra under non-turnover conditions displayed gradual reduction of the cyt c₅₅₂, following the step-wise decrease of electrode potential from 0.0 V to −0.6 V vs. standard calomel electrode (SCE). The spectral changes were fully reversible in both positive and negative direction of the scan potential, with average midpoint potential value of −0.42 V vs. SCE. Confocal microscopy analysis revealed that the thickness of biofilms under turnover conditions and non-turnover conditions was approximately 35 and 3.5 μm, respectively. This is the first study to observe the reversible redox conversion of cyt c₅₅₂ in viable G. sulfurreducens biofilms.