Controlled immobilisation of active enzymes on the cowpea mosaic virus capsid

Immobilisation of horseradish peroxidase (HRP) and glucose oxidase (GOX) via covalent attachment of modified enzyme carbohydrate to the exterior of the cowpea mosaic virus (CPMV) capsid gave high retention of enzymatic activity. The number of enzymes bound per virus was determined to be about eleven for HRP and 2-3 for GOX. This illustrates that relatively large biomacromolecules can be readily coupled to the virus surface using simple conjugation strategies. Virus-biomacromolecule hybrids have great potential for uses in catalysis, diagnostic assays or biosensors.     

Effects of Ketone Bodies on Brain Metabolism and Function in Neurodegenerative Diseases

Under normal physiological conditions the brain primarily utilizes glucose for ATP generation. However, in situations where glucose is sparse, e.g., during prolonged fasting, ketone bodies become an important energy source for the brain. The brain’s utilization of ketones seems to depend mainly on the concentration in the blood, thus many dietary approaches such as ketogenic diets, ingestion of ketogenic medium-chain fatty acids or exogenous ketones, facilitate significant changes in the brain’s metabolism. Therefore, these approaches may ameliorate the energy crisis in neurodegenerative diseases, which are characterized by a deterioration of the brain’s glucose metabolism, providing a therapeutic advantage in these diseases. Most clinical studies examining the neuroprotective role of ketone bodies have been conducted in patients with Alzheimer’s disease, where brain imaging studies support the notion of enhancing brain energy metabolism with ketones. Likewise, a few studies show modest functional improvements in patients with Parkinson’s disease and cognitive benefits in patients with—or at risk of—Alzheimer’s disease after ketogenic interventions. Here, we summarize current knowledge on how ketogenic interventions support brain metabolism and discuss the therapeutic role of ketones in neurodegenerative disease, emphasizing clinical data.     

GC–MS analysis of oxysterols and their formation in cultivated liver cells (HepG2)

Oxysterols play a key role in many (patho)physiological processes and they are potential biomarkers for oxidative stress in several diseases. Here we developed a rapid gas chromatographic-mass spectrometry-based method for the separation and quantification of 11 biologically relevant oxysterols bearing hydroxy, epoxy, and dihydroxy groups. Efficient chromatographic separation (resolution ≥ 1.9) was achieved using a medium polarity 35%-diphenyl/65%-dimethyl polysiloxane stationary phase material (30 m × 0.25 mm inner diameter and 0.25 μm film thickness). Based on thorough analysis of the fragmentation during electron ionization we developed a strategy to deduce structural information of the oxysterols. Optimized sample preparation includes (i) extraction with a mixture of n-hexane/iso-propanol, (ii) removal of cholesterol by solid phase extraction with unmodified silica, and (iii) trimethylsilylation. The method was successfully applied on the analysis of brain samples, showing consistent results with previous studies and a good intra- and interday precision of ≤20%. Finally, we used the method for the investigation of oxysterol formation during oxidative stress in HepG2 cells. Incubation with tert-butyl hydroperoxide led to a massive increase in free radical formed oxysterols (7-keto-chol > 7β-OH-chol >> 7α-OH-chol), while 24 h incubation with the glutathione peroxidase 4 inhibitor RSL3 showed no increase in oxidative stress based on the oxysterol pattern. Overall, the new method described here enables the robust analysis of a biologically meaningful pattern of oxysterols with high sensitivity and precision allowing us to gain new insights in the biological formation and role of oxysterols.     

Effect of processing conditions on the yield and failure response of an aliphatic polyketone terpolymer

The effect of processing conditions on the yield and failure behavior of an aliphatic polyketone terpolymer was studied. Testing and characterization were performed on samples that were extruded in the form of hollow cylinders. We performed the extrusion process at different shear rates and at different cooling rates to assess the effect that process conditions had on the polymer properties. We performed biaxial testing on the samples to characterize the failure envelopes, including the ductile–brittle transition condition for each process condition. The effect of shear rate was negligible, whereas the cooling rate significantly affected the failure behavior. To explain these differences in behavior, we performed characterization via differential scanning calorimetry, wide‐angle X‐ray diffraction, attenuated total reflection IR spectroscopy, dynamic mechanical thermal analysis, scanning electron microscopy, and residual stress measurements. A broad glass transition was found for all samples at temperatures higher than previously reported for this material. Alteration of the processing conditions did not influence the crystalline phase (percentage crystallinity, crystalline orientation, crystallite size, etc.). A change in spherulitic structure was also observed with altered cooling rate and is suggested to have contributed to the change in failure behavior. Residual stresses also affected the behavior of all samples. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 84: 318–334, 2002; DOI 10.1002/app.10334     

New phospholipid fatty acids from the marine spongeCinachyrella alloclada uliczka

The fatty acid composition of phospholipids from the Senegalese spongeCinachyrella alloclada was examined. Two new fatty acids not hitherto found in nature, namely 10,13-octadecadienoic acid and 16-tricosenoic acid, were identified. 8-Hexadecenoic, 13-nonadecenoic and 5,9,13-trimethyltretradecanoic fatty acids were also found for the first time in sponges. The latter compound (1.4% of the total fatty acid mixture), an isoprenoid fatty acid, accompanies the major fatty acid 4,8,12-trimethyltridecanoic acid (19.7%). The monomethyl branched fatty acids (22%) identified include 23-methylpentacosanoic acid (anteiso-26∶0), not previously observed in sponged. The major long-chain fatty acids encountered were the known 17-tetracosenoic 19-heptacosadienoic and 5,9,23-tricontatrienoic acid. Some sixty fatty acids were identified as methyl esters andN-acyl pyrrolidides by gas chromatography and gas chromatography/mass spectrometry.     

Surface chemistry and mechanical property changes of wood‐flour/high‐density‐polyethylene composites after accelerated weathering

Although wood–plastic composites have become more accepted and used in recent years and are promoted as low‐maintenance, high‐durability building products, they do experience a color change and a loss in mechanical properties with accelerated weathering. In this study, we attempted to characterize the modulus‐of‐elasticity (MOE) loss of photostabilized high‐density polyethylene (HDPE) and composites of wood flour and high‐density polyethylene (WF/HDPE) with accelerated weathering. We then examined how weathering changed the surface chemistry of the composites and looked at whether or not the surface changes were related to the MOE loss. By examining surface chemistry changes, we hoped to begin to understand what caused the weathering changes. The materials were left unstabilized or were stabilized with either an ultraviolet absorber or pigment. After 1000 and 2000 h of accelerated weathering, the samples were tested for MOE loss. Fourier transform infrared (FTIR) spectroscopy was employed to monitor carbonyl and vinyl group formation at the surface. Changes in the HDPE crystallinity were also determined with FTIR techniques. It was determined that structural changes in the samples (carbonyl group formation, terminal vinyl group formation, and crystallinity changes) could not be reliably used to predict changes in MOE with a simple linear relationship. This indicated that the effects of crosslinking, chain scission, and crystallinity changes due to ultraviolet exposure and interfacial degradation due to moisture exposure were interrelated factors for the weathering of HDPE and WF/HDPE composites. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 94: 2263–2273, 2004