In Vivo Inflammation Does Not Impair ABCA1-Mediated Cholesterol Efflux Capacity of HDL

HDL provides atheroprotection by facilitating cholesterol efflex from lipid-laden macrophages in the vessel wall. In vitro studies have suggested impaired efflux capacity of HDL following inflammatory changes. We assessed the impact of acute severe sepsis and mild chronic inflammatory disease on the efflux capacity of HDL. We hypothesize that a more severe inflammatory state leads to stronger impaired cholesterol efflux capacity. Using lipid-laden THP1 cells and fibroblasts we were able to show that efflux capacity of HDL from both patients with severe sepsis or with Crohn's disease (active or in remission), either isolated using density gradient ultracentrifugation or using apoB precipitation, was not impaired. Yet plasma levels of HDL cholesterol and apoA-I were markedly lower in patients with sepsis. Based on the current observations we conclude that inflammatory disease does not interfere with the capacity of HDL to mediate cholesterol efflux. Our findings do not lend support to the biological relevance of HDL function changes in vitro

CERTIFICATION REPORT: The certification of Amyloid β1-42 in CSF in ERM®-DA480/IFCC, ERM®-DA481/IFCC and ERM®-DA482/IFCC

This report describes the production of ERM®-DA480/IFCC, ERM®-DA481/IFCC and ERM®-DA482/IFCC, which are human cerebrospinal fluid (CSF) materials certified for the mass concentration of amyloid β1-42 peptide (Aβ1-42). These materials were produced by the European Commission, Joint Research Centre (EC-JRC) in collaboration with the International Federation for Clinical Chemistry and Laboratory Medicine (IFCC) following ISO Guide 34:2009 and are certified in accordance with ISO Guide 35:2006. The starting material used to prepare ERM-DA480/IFCC, ERM-DA481/IFCC and ERM-DA482/IFCC was human CSF collected from normal pressure hydrocephalus patients by continuous lumbar drainage. After collection, the CSF was aliquoted and frozen at -80 °C. For the preparation of each certified reference material (CRM) a selected number of CSF donations were thawed, pooled, mixed, filled in microvials and stored at (-70 ± 10) °C immediately thereafter. Between unit-homogeneity was quantified and stability during dispatch and storage were assessed in accordance with ISO Guide 35:2006 [ ]. The material was characterised by an interlaboratory comparison of laboratories of demonstrated competence and adhering to ISO/IEC 17025 [ ]. Technically invalid results were removed but no outlier was eliminated on statistical grounds only. Uncertainties of the certified values were calculated in accordance with the Guide to the Expression of Uncertainty in Measurement (GUM) [ ] and include uncertainties related to possible inhomogeneity, instability and characterisation. The materials are intended for the calibration of methods, quality control and/or the assessment of method performance. As with any reference material, they can be used for establishing control charts or validation studies. The CRMs are available in microvials containing at least 0.5 mL of frozen liquid. The minimum amount of sample to be used is 15 µL.JRC.F.6-Reference Material

Reduced CETP glycosylation and activity in patients with homozygous B4GALT1 mutations

The importance of protein glycosylation in regulating lipid metabolism is becoming increasingly apparent. We set out to further investigate this by studying the effects of defective glycosylation on plasma lipids in patients with B4GALT1-CDG, caused by a mutation in B4GALT1 with defective N-linked glycosylation. We studied plasma lipids, cholesteryl ester transfer protein (CETP) glyco-isoforms with isoelectric focusing followed by a western blot and CETP activity in three known B4GALT1-CDG patients and compared them with 11 age- and gender-matched, healthy controls. B4GALT1-CDG patients have significantly lowered non-high density lipoprotein cholesterol (HDL-c) and total cholesterol to HDL-c ratio compared with controls and larger HDL particles. Plasma CETP was hypoglycosylated and less active in B4GALT1-CDG patients compared to matched controls. Our study provides insight into the role of protein glycosylation in human lipoprotein homeostasis. The hypogalactosylated, hypo-active CETP found in patients with B4GALT1-CDG indicates a role of protein galactosylation in regulating plasma HDL and LDL. Patients with B4GALT1-CDG have large HDL particles probably due to hypogalactosylated, hypo-active CETP

Increased masticatory activity and quality of life in elderly persons with dementia-a longitudinal matched cluster randomized single-blind multicenter intervention study.

Background: Worldwide, millions of people are suffering from dementia and this number is rising. An index of quality of life (QoL) can describe the impact a disease or treatment has on a person's wellbeing. QoL comprises many variables, including physical health and function, and mental health and function. QoL is related to masticatory ability and physical activity. Animal studies show that disruption of mastication due to loss of teeth or a soft diet leads to memory loss and learning problems. Since these are common complaints in dementia, it is hypothesized that improvement of masticatory function and normalization of diet consistency can increase QoL in elderly persons suffering from dementia. Therefore, the goal of the present study is to examine whether an increase in masticatory activity, achieved by increased food consistency and enhancement of masticatory function through improved oral health care has a positive effect on QoL, including cognition, mood, activities of daily living (ADL), and circadian rhythm in elderly persons with dementia.Methods and design: The described study is a prospective longitudinal matched cluster randomized single-blind multicenter study. Participants are elderly persons living in the Netherlands, suffering from dementia and receiving psychogeriatric care. An intervention group will receive improved oral health care and a diet of increased consistency. A control group receives care as usual. Participants will be assessed four times; outcome variables besides QoL are cognition, mood, independence, rest-activity rhythm, blood pressure, and masticatory function.Discussion: This research protocol investigates the effect of an intervention executed by daily caregivers. The intervention will increase masticatory activity, which is achieved by three different actions, (providing oral health care, increasing food consistency, or a combination of both). There is a certain amount of variety in the nature of the interventions due to local differences in nursing homes. This might be a scientific weakness in the study design; however, a practical implementation of any findings will be subject to the same factors, making this study design clinically relevant.Trial registration: NTR1561. © 2013 Weijenberg et al.; licensee BioMed Central Ltd

Mapping lightning in the sky with a mini array

Mini arrays are commonly used for infrasonic and seismic studies. Here we report for the first time the detection and mapping of distant lightning discharges in the sky with a mini array. The array has a baseline to wavelength ratio $\sim$4.2 ${ \cdot}$ $10^{-2}$ to record very low frequency electromagnetic waves from 2 to 18 kHz. It is found that the mini array detects $\sim$69 lightning pulses per second from cloud-to-ground and in-cloud discharges, even though the parent thunderstorms are $\sim$900-1100 km away and a rigorous selection criterion based on the quality of the wavefront across the array is used. In particular, lightning pulses that exhibit a clockwise phase progression are found at larger elevation angles in the sky as the result of a birefringent subionospheric wave propagation attributed to ordinary and extraordinary waves. These results imply that long range lightning detection networks might benefit from an exploration of the wave propagation conditions with mini arrays

Formulation, quality control and shelf life of the experimental cytostatic drug cyclopentenyl cytosine

This paper describes the formulation and quality control of an aqueous sterilized formulation of the experimental cytostatic drug cyclopentenyl cytosine (CPEC) to be used in Phase I/II clinical trials. The raw drug substance was extensively tested. A High Pressure Liquid Chromotography (HPLC) method was validated for the quality control of the formulated product. The aqueous formulation was found to be stable for at least 2 years at 2-8 degrees C. Sterilization (15 min at 121 degrees C) showed no influence on drug stability. The results show that CPEC can be formulated in an aqueous solution. The described HPLC method is a useful tool in the pharmaceutical quality contro

Nanotwinned silver nanowires: Structure and mechanical properties

Ag nanowires have been manufactured with the ultrathin layer electrochemical deposition method. Transmission electron microscopy revealed a multiple twinned nanostructure that has not been reported in comparable structures using a non-template self-assembling preparation method. The {1 1 1} twin planes are oriented parallel to the wire axis. The influence of the twin boundaries on the elastic and plastic properties of these wires has been investigated. Atomistic calculations revealed no influence of the twin boundaries on the Young's modulus of the wire for this geometry and along the direction. However, the twin boundaries exhibited a strong orientation-dependent influence on the plastic deformation when indented orthogonal ( direction) or parallel ( direction) to the twin boundaries, reflecting the interaction between the glide systems active in fcc metals and the stress direction. Heating experiments showed a high heat resistance for the twin boundaries till sublimation in an in situ TEM heating experiment. Sublimation followed the twin boundaries, indicating the lower binding energy of hcp stacking in comparison to fcc stacking. (C) 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved

CSF Aβ1-42 – an excellent but complicated Alzheimer's biomarker – a route to standardisation

The 42 amino acid form of amyloid β (Aβ1-42) in cerebrospinal fluid (CSF) has been widely accepted as a central biomarker for Alzheimer's disease. Several immunoassays for CSF Aβ1-42 are commercially available, but can suffer from between laboratory and batch-to-batch variability as well as lack of standardisation across assays. As a consequence, no general cut-off values have been established for a specific context of use (e.g., clinical diagnostics) and selection of individuals for enrolment in clinical trials (patient stratification) remains challenging. The International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) has initiated a working group for CSF proteins (WG-CSF) to facilitate standardisation of CSF Aβ1-42 measurement results. The efforts of the IFCC WG-CSF include the development of certified reference materials (CRMs) and reference measurements procedures (RMPs) for key biomarkers. Two candidate RMPs for quantification of Aβ1-42 in CSF based on liquid chromatography tandem mass spectrometry have been developed and tested in two ring trials. Furthermore, two commutability studies including native CSF pools, artificial CSF and spiked materials have been completed. On the basis of these studies, human CSF pools containing only endogenous Aβ1-42 at three concentrations were selected as the format for future CRMs that are now being processed.JRC.F.6-Reference Material

High prevalence of mutations in LCAT in patients with low HDL cholesterol levels in The Netherlands: identification and characterization of eight novel mutations

Lecithin:cholesterol acyltransferase (LCAT) is crucial to the maturation of high-density lipoprotein (HDL). Homozygosity for LCAT mutations underlies rare disorders characterized by HDL-cholesterol (HDL-c) deficiency while heterozygotes have half normal HDL-c levels. We studied the prevalence of LCAT mutations in referred patients with low HDL-c to better understand the molecular basis of low HDL-c in our patients. LCAT was sequenced in 98 patients referred for HDL-c T (p.E134D), c.403T>A (p.Y135N), c.964C>T (p.R322C), c.296G>C (p.W99S), c.736G>T (p.V246F), c.802C>T (p.R268C), c.945G>A (p.W315X), c.1012C>T (p.L338F), and c.1039C>T (p.R347C)--were shown to be functional through in vitro characterization. The effect of several mutations on the core protein structure was studied by a three-dimensional (3D) model. Unlike previous reports, functional mutations in LCAT were found in 29% of patients with low HDL-c, thus constituting a common cause of low HDL-c in referred patients in The Netherland