Stimulation of cholesteryl ester synthesis in human monocyte-derived macrophages by low-density lipoproteins from type 1 (insulin-dependent) diabetic patients:the influence of non-enzymatic glycosylation of low-density lipoproteins

Diabetes mellitus is an independent risk factor in the development of atherosclerosis. In this study we aimed to demonstrate whether there is an abnormal interaction between low-density lipoproteins from diabetic patients and human macrophages. We measured cholesteryl ester synthesis and cholesteryl ester accumulation in human monocyte-derived macrophages (obtained from non-diabetic donors) incubated with low density lipoproteins from Type 1 (insulin-dependent) diabetic patients in good or fair glycaemic control. Low density lipoproteins from the diabetic patients stimulated more cholesteryl ester synthesis than low density lipoproteins from non-diabetic control subjects (7.19 +/- 1.19 vs 6.11 +/- 0.94 nmol/mg cell protein/20 h, mean +/- SEM, p less than 0.05). The stimulation of cholesteryl ester synthesis by low density lipoproteins isolated from diabetic patients was paralleled by a significant increase in intracellular cholesteryl ester accumulation (p less than 0.02). There were no significant differences in the lipid composition of low density lipoproteins between the diabetic and control groups. Non-enzymatic glycosylation of low density lipoproteins was higher in the diabetic group (p less than 0.01) and correlated significantly with cholesteryl ester synthesis (r = 0.58). Similarly, low-density lipoproteins obtained from non-diabetic subjects and glycosylated in vitro stimulated more cholesteryl ester synthesis in macrophages than control low density lipoproteins. The increase in cholesteryl ester synthesis and accumulation by cells exposed to low density lipoproteins from diabetic patients seems to be mediated by an increased uptake of these lipoproteins by macrophages.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of in vitro non-enzymatic glycosylation of human skin collagen on susceptibility to collagenase digestion

The effect of glycosylation on susceptibility of skin collagen to collagenase digestion was studied in a skin sample obtained at autopsy from the interscapular region of a 24 year old white male who had died of an acute illness and who had no history of diabetes. Homogeneous suspensions of insoluble collagen were prepared, and were incubated in 50 mmol l-1 dextrose at pH 7.35 and 37 degrees C for 7 days. Non-enzymatic glycosylation measured by the weak acid hydrolysis/thiobarbituric acid method increased from 13.1 +/- 1.0 (n = 5) to 45.2 +/- 5.5 (n = 8) nmol fructose per 10 mg collagen (P less than 0.001). Digestion of collagen using clostridial collagenase was monitored by measuring (a) hydroxyproline content and (b) absorption at 206 nm of the supernatant after centrifugation to remove substrate. The rate of digestion was similar in glycosylated and control collagen. We conclude that the ketoamine link formed in non-enzymatic glycosylation does not increase the resistance of collagen to enzymatic digestion. The possibility remains that subsequent rearrangement of this link could be important in this respect.

Non-enzymatic glycosylation of skin collagen in patients with type 1 (insulin-dependent) diabetes mellitus and limited joint mobility

Forearm skin biopsies were obtained from diabetic subjects with and without limited joint mobility, and from non-diabetic control subjects. Collagen purified from these samples was assayed for non-enzymatic glycosylation. The level in all diabetic patients was significantly greater than that in control subjects (p less than 0.001), but those diabetic patients with limited joint mobility had a level of collagen glycosylation similar to that in those with normal joints (15.3 +/- 1.3 and 16.5 +/- 1.3 nmol fructose/10 mg protein, respectively; mean +/- SEM). Glycosylation of collagen in the diabetic patients correlated with glycosylated haemoglobin measured at the time of skin biopsy (r = 0.60). These results do not support the hypothesis that non-enzymatic glycosylation of collagen, as reflected by the ketoamine link, plays an important role in the development of limited joint mobility in diabetes.

Predicting critical source areas of sediment in headwater catchments

Mitigation of diffuse nutrient and sediment delivery to streams requires successful identification andmanagement of critical source areas within catchments. Approaches to predicting high risk areas forsediment loss have typically relied on structural drivers of connectivity and risk, with little considera-tion given to process driven water quality responses. To assess the applicability of structural metrics topredict critical source areas, geochemical tracing of land use sources was conducted in three headwateragricultural catchments in Co. Down and Co. Louth, Ireland, within a Monte Carlo framework. Outputswere applied to the inverse optimisation of a connectivity model, based on LiDAR DEM data, to assess theefficacy of land use risk weightings to predict sediment source contributions over the 18 month studyperiod in the Louth Upper, Louth Lower and Down catchments. Results of the study indicated sedimentproportions over the study period varied from 6 to 10%, 84 to 87%, 4%, and 2 to 3% for the Down Catch-ment, 79 to 85%, 9 to 17%, 1 to 3% and 2 to 3% in the Louth Upper and 2 to 3%, 79 to 85%, 10 to 17%and 2 to 3% in the Louth Lower for arable, channel bank, grassland, and woodland sources, respectively.Optimised land use risk weightings for each sampling period showed that at the larger catchment scale,no variation in median land use weightings were required to predict land use contributions. However,for the two smaller study catchments, variation in median risk weightings was considerable, which mayindicate the importance of functional connectivity processes at this spatial scale. In all instances, arableland consistently generated the highest risk of sediment loss across all catchments and sampling times.This study documents some of the first data on sediment provenance in Ireland and indicates the needfor cautious consideration of land use as a tool to predict critical source areas at the headwater scale

Halophilic microorganisms as a source of novel antibiofilm compounds

Objectives: There is great urgency for alternate sources of antibiotics to be identified. One relatively untapped source of novel bioproducts, including antimicrobials, is organisms derived from extreme environments. Halophiles (which require high salt concentrations) are one such group which is being increasingly explored for their biotechnological potential. The aim of this study was to identify halophilic environmental isolates which possessed in vitro and in vivo antimicrobial and antibiofilm activities. Methods: 73 halophilic bacteria and archaea were isolated from Kilroot salt mine in Northern Ireland. Culture extracts of each isolate were screened for antimicrobial and antibiofilm activity against numerous pathogenic bacteria, including Staphylococcus species and Pseudomonas aeruginosa, both model strains and clinical isolates. The methods used included disc diffusion assays of crude extracts, MIC screening, the MBEC assay, and an in vivo model based on the Greater Wax Moth (Galleria mellonella). Results: The assays indicated >50% of extracts displayed antimicrobial and antibiofilm activity against at least one pathogen, the majority being Staphylococcus species, but also E. coli and P. aeruginosa. Biofilms were either reduced or eradicated by halophile extracts when tested with the MBEC device. Further experiments demonstrated that these effects could be replicated in vivo, with extracts reducing the severity of infections and enhancing the survival of infected G. mellonella. Conclusions: The importance of extremophiles to pharmaceutical research should not be underestimated. While not yet fully characterised, based on the data obtained, the halophiles isolated during this study may provide a promising reservoir of novel antimicrobial and antibiofilm compounds.

Power allocation strategies across N orthogonal channels for single-relay networks at both source and relay

We consider a wireless relay network with one source, one relay and one destination, where communications between nodes are preformed over N orthogonal channels. This, for example, is the case when orthogonal frequency division multiplexing is employed for data communications. Since the power available at the source and relay is limited, we study optimal power allocation strategies at the source and relay in order to maximize the overall source-destination capacity. Depending on the availability of the channel state information at both the source and relay or only at the relay, power allocation is performed at both the source and relay or only at the relay. Considering different setups for the problem, various optimization problems are formulated and solved. Some properties of the optimal solution are also proved.

Physical and numerical constraints in source modeling for finite difference simulation of room acoustics

In finite difference time domain simulation of room acoustics, source functions are subject to various constraints. These depend on the way sources are injected into the grid and on the chosen parameters of the numerical scheme being used. This paper addresses the issue of selecting and designing sources for finite difference simulation, by first reviewing associated aims and constraints, and evaluating existing source models against these criteria. The process of exciting a model is generalized by introducing a system of three cascaded filters, respectively, characterizing the driving pulse, the source mechanics, and the injection of the resulting source function into the grid. It is shown that hard, soft, and transparent sources can be seen as special cases within this unified approach. Starting from the mechanics of a small pulsating sphere, a parametric source model is formulated by specifying suitable filters. This physically constrained source model is numerically consistent, does not scatter incoming waves, and is free from zero- and low-frequency artifacts. Simulation results are employed for comparison with existing source formulations in terms of meeting the spectral and temporal requirements on the outward propagating wave.

Table-Top Laser-Based Source of Femtosecond, Collimated, Ultrarelativistic Positron Beams

The generation of ultrarelativistic positron beams with short duration (τ_e⁺≃30 fs), small divergence (θ_e⁺≃3 mrad), and high density (n _e⁺≃1014-1015 cm^-3) from a fully optical setup is reported. The detected positron beam propagates with a high-density electron beam and γ rays of similar spectral shape and peak energy, thus closely resembling the structure of an astrophysical leptonic jet. It is envisaged that this experimental evidence, besides the intrinsic relevance to laser-driven particle acceleration, may open the pathway for the small-scale study of astrophysical leptonic jets in the laboratory.