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.     

The effect of microstructure on hydrogen permeability of high strength steels

Hydrogen diffusivity and trapping have been studied in two advanced high strength steel grades and model samples using electrochemical permeation test. Microstructures of CP1000 and DP1000 steels consist of ferrite, martensite and a small fraction of retained austenite. In addition, bainite is present in CP1000. Model phases with predominance of a particular phase have been prepared by specific heat treatment. DP1000 has shown the lowest diffusivity among all materials, while ferritic model sample has shown the highest. Differences in hydrogen diffusion coefficient values are linked to trapping microstructural characteristics and grain size.     

Influence of the Cooling Temperature on the Mechanical Behavior of an Oxidized Self-Healing Ceramic Matrix Composite

Composites made of ceramic for both fibers and matrix are aimed at being used in aerospace applications, which means being submitted to mechanical stresses at high temperatures in oxidizing and corrosive environments for long durations. The oxidation/corrosion resistance of these materials is linked to their ability to self-heal by creating an oxide phase which can restrict the access of the oxygen into the bulk of the material. During thermomechanical cycles, the distribution of O₂ inside the material as well as the distribution and the state of the liquid oxides will undergo some modifications. A specific test has been conceived and performed in order to evidence the influence of the viscosity of this oxide on the mechanical properties of the material. Results have shown that the stiffening which is observed seems to be proportional to the increase in the viscosity of the sealing oxide, in conjunction with the decrease in the temperature.     

Effects of the mycotoxin deoxynivalenol on human primary hepatocytes

Toxic effects of the mycotoxin deoxynivalenol (DON) observed in animals range from diarrhea, vomiting, gastro-intestinal inflammation to necrosis of several tissues. In the last years, DON has been tested in hepatocytes of several animal species for its cytotoxicity. However, these tests are limited to the use of animal cells. No studies using human hepatocytes are available. Further investigations with the human hepatocellular liver carcinoma cell line HepG2 might be limited due to the disadvantages of cell lines (e. g. immortalization, tumor derivation, longtime cultivation) and do not necessarily reflect the response of normal human cells. In order to overcome this problem and to be closer to the human situation, we studied the effect of DON in human primary hepatocytes and compared these data to the effects in the HepG2 cell line. Cell viability, apoptotic and necrotic cell death, albumin secretion and metabolic activity were determined. It could be demonstrated that DON has a distinct cytotoxic effect on human primary hepatocytes. Viability, protein content and albumin secretion were reduced in a dose-dependent manner. The apoptotic key enzyme caspase-3 was activated, while LDH release occurred only after long incubation time due to a secondary necrosis. Furthermore, we studied the metabolism of DON using LC-MS/MS. DON was neither metabolized by primary hepatocytes cells nor by the HepG2 cell line.     

Validation of a diatom-based index of water quality confirms its utility in monitoring of the Lake Erie's nearshore area

We conducted a validation of the Planktonic Diatom Index (PDI) to demonstrate the utility of a water quality index for the monitoring of Lake Erie's nearshore pelagial zone. Using a large, independent dataset from the Western and Central Basins of Lake Erie for validation ensures realistic assessment of the performance of the index. Diatom-based biomonitoring allows for the inference of integrative information about water quality based on diatom species composition. The PDI is based on the assumption that phosphorus, an established proxy for eutrophication, is instrumental in the structuring of diatom communities. In this study, PDI scores and measured total phosphorus were significantly correlated (r²?=?0.34, r²?=?0.63 outliers removed). However, when samples were considered on a basin-wide basis, the PDI scores were not significantly predicted by measured total phosphorus in the Western Basin. We suggest that snapshot phosphorus measurements are less likely to represent the overall condition in the highly variable, eutrophic Western Basin. When multiple phosphorus measurements were averaged over time, the relationship with the integrative PDI scores was more apparent (r²?=?0.52). Through validation with an independent dataset, we show that the PDI is likely a monitoring tool that provides a robust assessment of water quality in the pelagial zone of the nearshore waters in Lake Erie.     

The effect of mayfly (Hexagenia spp.) burrowing activity on sediment oxygen demand in western Lake Erie

Previous studies support the hypothesis that large numbers of infaunal burrow-irrigating organisms in the western basin of Lake Erie may increase significantly the sediment oxygen demand, thus enhancing the rate of hypolimnetic oxygen depletion. We conducted laboratory experiments to quantify burrow oxygen dynamics and increased oxygen demand resulting from burrow irrigation using two different year classes of Hexagenia spp. nymphs from western Lake Erie during summer, 2006. Using oxygen microelectrodes and hot film anemometry, we simultaneously determined oxygen concentrations and burrow water flow velocities. Burrow oxygen depletion rates ranged from 21.7 mg/nymph/mo for 15 mm nymphs at 23 °C to 240.7 mg/nymph/mo for 23 mm nymphs at 13 °C. Sealed microcosm experiments demonstrated that mayflies increase the rate of oxygen depletion by 2–5 times that of controls, depending on size of nymph and water temperature, with colder waters having greater impact. At natural population densities, nymph pumping activity increased total sediment oxygen demand 0.3–2.5 times compared to sediments with no mayflies and accounted for 22–71% of the total sediment oxygen demand. Extrapolating laboratory results to the natural system suggest that Hexagenia spp. populations may exert a significant control on oxygen depletion during intermittent stratification. This finding may help explain some of the fluctuations in Hexagenia spp. population densities in western Lake Erie and suggests that mayflies, by causing their own population collapse irrespective of other environmental conditions, may need longer term averages when used as a bio-indicator of the success of pollution-abatement programs in western Lake Erie and possibly throughout the Great Lakes.     

Application of Motor Control Centers to Systems Having High Available Fault Currents

Rising costs of manufacturing and construction have led to the development of modern distribution systems having higher kVA ratings with larger available fault currents. Because of this, it is essential that special considerations be given to methods that will reduce the available fault current at the motor-control-center incoming terminals to a value that is within the rating of the motor control center.     

Breeding birds and anurans of dynamic coastal wetlands in Green Bay,Lake Michigan

Breeding birds and anurans (frogs and toads) in coastal wetlands of Green Bay, Lake Michigan vary dynamically with changing water levels, habitat type, and geography. We describe species assemblages over a seven-year period (2011–2017) beginning with historic low water levels followed by an increase in average lake level of 0.85?m. In general, species richness and abundance of marsh-obligate species responded positively to increasing water levels, although several species of shallow wetlands (sandhill crane, sedge wren, swamp sparrow, and American toad) showed the opposite trend. Anuran assemblages were more diverse in the middle and upper bay, coinciding with a well-established nutrient gradient from the hypereutrophic lower bay to more oligotrophic waters of the upper bay. Three marsh-obligate bird species (black tern, sandhill crane, and sedge wren) were significantly more abundant in the middle or upper bay while sora, American coot, and common gallinule were more abundant in the eutrophic lower bay. Our findings have several important implications for conservation. Inland wetlands near the coast (including diked wetlands) might play a key ecological role by providing refugia for some species during low water years. However, the importance of shallow coastal wetlands and nearshore gradients of wetland habitat might be overlooked during low water years; when high water returns, these areas can become extremely productive and species-rich.     

Analysis of temperature coefficients for III–V multi‐junction concentrator cells

The temperature dependence of the I–V parameters of different III–V multi‐junction concentrator cells at several concentration levels was investigated. Moreover, the influence of spectral changes on the temperature coefficients of multi‐junction solar cells was examined. Complete sets of temperature coefficients of a metamorphic Ga_0.35In_0.65P/Ga_0.83In_0.17As dual‐junction cell, a metamorphic Ga_0.35In_0.65P/Ga_0.83In_0.17As/Ge triple‐junction cell and a lattice‐matched Ga_0.50In_0.50P/Ga_0.99In_0.01As/Ge triple‐junction cell determined under well‐controlled laboratory conditions are reported. Copyright © 2012 John Wiley & Sons, Ltd.     

Biomimetic hydroxyapatite coating on glass coverslips for the assay of osteoclast activity in vitro

The osteoclast (OC) is the cell type responsible for the resorption of bone. The activity of this cell is important in the aetiology of a large number of skeletal pathologies, and also for the biocompatibility and osseointegration of orthopaedic implant materials. OC mediated acid hydrolysis of calcium phosphate from the bone matrix offers a prime means of studying the biology and activity of this cell type. We have developed a method of coating glass coverslips with a hydroxyapatite (HA)-like mineral, using a biomimetic approach. Hydroxylation followed by formation of a self assembled monolayer (SAM) using the surfactant triethoxysilylpropyl succinic anhydride (TESPSA), allowed biomimetic deposition of HA-like mineral from a simulated body fluid (SBF). The biocompatibility of the TESPSA SAM-HA coated glass coverslips was tested by culturing human mature OC present in samples of giant cell tumour of bone (GCT). Parameters of OC activity were assayed, including F-actin ring formation, release of calcium and formation of osteoclastic resorption pits, confirming that OC were able to attach to and resorb the coated surface. This approach for the preparation of HA coatings on glass coverslips could have wide applicability for the study of osteoclast behaviour in vitro. Gerald J. Atkins and Peter Majewski share senior author status.