Signal enhancement of surface plasmon resonance immunoassay using enzyme precipitation-functionalized gold nanoparticles: A femto molar level measurement of anti-glutamic acid decarboxylase antibody

Colloidal gold nanoparticles (AuNPs) and precipitation of an insoluble product formed by HRP-biocatalyzed oxidation of 3,3'-diaminobenzidine (DAB) in the presence of H2O2 were used to enhance the signal obtained from the surface plasmon resonance (SPR) biosensor. The AuNPs were synthesized and functionalized with HS-OEG(3)-COOH by self assembling technique. Thereafter, the HS-OEG3-COOH functionalized nanoparticles were covalently conjugated with horseradish peroxidase (HRP) and anti IgG antibody to form an enzyme-immunogold complex. Characterizations were performed by several methods: UV-vis absorption, DLS, HR-TEM and Fr-IR. The Au-anti IgG-HRP complex has been applied in enhancement of SPR immunoassay using a sensor chip constructed by 1:9 molar ratio of HS-OEG(6)-COOH and HS-OEG(3)-OH for detection of anti-GAD antibody. As a result, AuNPs showed their enhancement as being consistent with other previous studies while the enzyme precipitation using DAB substrate was applied for the first time and greatly amplified the SPR detection. The limit of detection was found as low as 0.03 ng/ml of anti-GAD antibody (or 200 fM) which is much higher than that of previous reports. This study indicates another way to enhance SPR measurement, and it is generally applicable to other SPR-based immunoassays.

Amplitude modulated signal transmission through a phase conjugating lens

Free space transmission of an on-off modulated sinusoidal signal through a phase conjugating lens (PCL) is theoretically examined using a combined time/frequency domain approach. The on-off keyed (OOK) signal is generated by a dipole antenna located in the far-field zone of the lens. The PCL consists of a dual layer of antenna elements interconnected via phase conjugating circuitry. We demonstrate that electromagnetic interference between antenna elements creates spatially localised areas of good-quality reception and zones where the signal is significantly denigrated by interference. Next, it is shown that destructive interference and packet desynchronisation effects critically depend on bit rate. It is also shown that a circular concave lens can be used to produce high-quality signal reception in a given direction while suppressing signal reception in all other directions. The effect that the bandwidth of the phase conjugating unit has on the transmitted signal properties for the cases of high and low bit rate OOK modulation are studied and a signal quality characterisation scheme is proposed which uses cross-correlation. The results of the study yields understanding of the performance of phase conjugating arrays under OOK modulation. The work suggests a novel approach for realising a secure communication wireless system.

Native and modified LDL activate extracellular signal-regulated kinases in mesangial cells

Glycation and/or oxidation of LDL may promote diabetic nephropathy. The mitogen-activated protein kinase (MAPK) cascade, which includes extracellular signal-regulated protein kinases (ERKs), modulates cell function. Therefore, we examined the effects of LDL on ERK phosphorylation in cultured rat mesangial cells. In cells exposed to 100 microg/ml native LDL or LDL modified by glycation, and/or mild or marked (copper-mediated) oxidation, ERK activation peaked at 5 min. Five minutes of exposure to 10-100 microg/ml native or modified LDL produced a concentration-dependent (up to sevenfold) increase in ERK activity. Also, 10 microg/ml native LDL and mildly modified LDL (glycated and/or mildly oxidized) produced significantly greater ERK activation than that induced by copper-oxidized LDL +/- glycation (P <0.05). Pretreatment of cells with Src kinase and MAPK kinase inhibitors blocked ERK activation by 50-80% (P <0.05). Native and mildly modified LDL, which are recognized by the native LDL receptor, induced a transient spike of intracellular calcium. Copper-oxidized (+/- glycation) LDL, recognized by the scavenger receptor, induced a sustained rise in intracellular calcium. The intracellular calcium chelator (EGTA/AM) further increased ERK activation by native and mildly modified LDL (P <0.05). These findings demonstrate that native and modified LDL activate ERKs 1 and 2, an early mitogenic signal, in mesangial cells and provide evidence for a potential link between modified LDL and the development of glomerular injury in diabetes.

Hemostatic alterations with exercise conditioning in NIDDM

Various parameters of coagulation and fibrinolysis were measured in 13 men (aged 54 +/- 3 yr) with non-insulin-dependent diabetes mellitus (NIDDM) before and after 12-14 wk of exercise training. Subjects exercised for 30 min 3 times/wk at 70% of maximum O2 consumption (VO2max). Training increased VO2max by 12.5% but did not alter body weight, relative body fat, blood pressure, cholesterol, triglycerides, or high-density lipoprotein cholesterol. Slight downward trends were apparent for fasting glucose and insulin, but glycosylated hemoglobin was unchanged. There were no changes in coagulation parameters of plasminogen, hematocrit, or alpha 2-antiplasmin. Plasma fibrinogen (303 +/- 24.2 vs. 256 +/- 12.3 mg/dl) and fibronectin (380 +/- 41.9 vs. 301 +/- 22.2 micrograms/ml) were significantly reduced (P less than 0.02) by exercise conditioning. Three assays of fibrinolytic activity (tissue plasminogen activator, euglobulin lysis time, and an isotopic measure of fibrinolysis) confirmed that neither basal fibrinolysis nor the fibrinolytic responses to venous occlusion and maximal exercise were significantly altered. Exercise conditioning may have antithrombotic effects in NIDDM by reducing plasma fibrinogen and fibronectin. Although the significance of the fall in fibronectin awaits further studies, the reduction in plasma fibrinogen gives a rationale for the use of exercise training in men with NIDDM.

Sirolimus Stimulates Vascular Stem/Progenitor Cell Migration and Differentiation Into Smooth Muscle Cells via Epidermal Growth Factor Receptor/Extracellular Signal-Regulated Kinase/β-Catenin Signaling Pathway

Sirolimus-eluting stent therapy has achieved considerable success in overcoming coronary artery restenosis. However, there remain a large number of patients presenting with restenosis after the treatment, and the source of its persistence remains unclarified. Although recent evidence supports the contribution of vascular stem/progenitor cells in restenosis formation, their functional and molecular responses to sirolimus are largely unknown.

Analysis of Load Impact on Small-signal Stability based on Load Active Power Sensitivity

While load flow conditions vary with different loads, the small-signal stability of the entire system is closely related with to the locations, capacities and models of loads. In this paper, load impacts with different capacities and models on the small-signal stability are analysed. In the real large-scale power system case, the load sensitivity which denotes the sensitivity of the eigenvalue with respect to the load active power is introduced and applied to rank the loads. The loads with high sensitivity are also considered.

A Wavelet Approach to the Selective Computation of Eigenvalues for Small Signal Stability Analysis

This paper proposes a method to assess the small signal stability of a power system network by selective determination of the modal eigenvalues. This uses an accelerating polynomial transform, designed using approximate eigenvalues
obtained from a wavelet approximation. Application to the IEEE 14 bus network model produced computational savings of 20%,over the QR algorithm.

Myeloid cells expressing VEGF and arginase-1 following uptake of damaged retinal pigment epithelium suggests potential mechanism that drives the onset of choroidal angiogenesis in mice

Whilst data recognise both myeloid cell accumulation during choroidal neovascularisation (CNV) as well as complement activation, none of the data has presented a clear explanation for the angiogenic drive that promotes pathological angiogenesis. One possibility that is a pre-eminent drive is a specific and early conditioning and activation of the myeloid cell infiltrate. Using a laser-induced CNV murine model, we have identified that disruption of retinal pigment epithelium (RPE) and Bruch's membrane resulted in an early recruitment of macrophages derived from monocytes and microglia, prior to angiogenesis and contemporaneous with lesional complement activation. Early recruited CD11b(+) cells expressed a definitive gene signature of selective inflammatory mediators particularly a pronounced Arg-1 expression. Accumulating macrophages from retina and peripheral blood were activated at the site of injury, displaying enhanced VEGF expression, and notably prior to exaggerated VEGF expression from RPE, or earliest stages of angiogenesis. All of these initial events, including distinct VEGF (+) Arg-1(+) myeloid cells, subsided when CNV was established and at the time RPE-VEGF expression was maximal. Depletion of inflammatory CCR2-positive monocytes confirmed origin of infiltrating monocyte Arg-1 expression, as following depletion Arg-1 signal was lost and CNV suppressed. Furthermore, our in vitro data supported a myeloid cell uptake of damaged RPE or its derivatives as a mechanism generating VEGF (+) Arg-1(+) phenotype in vivo. Our results reveal a potential early driver initiating angiogenesis via myeloid-derived VEGF drive following uptake of damaged RPE and deliver an explanation of why CNV develops during any of the stages of macular degeneration and can be explored further for therapeutic gain.

Soil conditioning for EPBMs

Nowadays, the Earth Pressure Balanced (EPB) shields are used more and more caused by both the mechanical development of the machines and the more effective use of additives such as foams and polymers. These additives are used to condition spoil in the screw conveyor. A crucial component of EPB tunnelling applications is ground conditioning. Thus, in order to permit quantitative measurements of the conditioned ground behavior, it is necessary to develop and use a test able to simulate the extraction of earth from the chamber with a screw conveyor. This reliable test procedure can provide quantitative parameters that can really describe the conditioned soil behavior.

Laboratory tests for EPB tunnelling soil conditioning

The screw conveyor system plays a fundamental role during the EPB tunnelling operations for the tunnel face pressure control. On the other hand, the use of additives such chemical foams is even more applied in order to extend the EPB technology to the cohesionless soils. Despite the extensive use of the EPB technique in urban environment, little knowledge exists in the understanding of the behavior of such conditioned soil during the excavation operations. At the Turin University of Technology the Tunnelling and Underground Space Centre, in the mainframe of a wider research on soil conditioning, has developed an experimental apparatus that simulates the extraction phase with screw conveyor from a pressurized tank. In this paper the apparatus is presented and the results of a first series of tests carried out on sand are discussed. © 2007 Taylor & Francis Group.

Soil conditioning of sand for EPB applications:A laboratory research

EPB tunnelling requires the application of soil conditioning to increase its field of applicability particularly for cohesionless soils. Choosing the most suitable conditioning set for the various soils requires the use of a feasible laboratory test which can permit to define the characteristics of the conditioned soils and provide measurable data. A series of tests has been carried out using a laboratory screw conveyor device which was designed for this purpose and which simulates the extraction of the spoil from a pressure chamber in a similar way as in EPB tunnelling. The tested soils were medium-grain sands with varying amounts of silt and the tested conditioned mixtures were obtained with different water contents and amounts of foam. A simple slump test was also used to analyze the global characteristics of the conditioned soils. The test has shown that the proposed laboratory procedure permits a quantitative comparison to be made between different conditioning amounts and agents on the basis of measurable parameters. © 2007 Elsevier Ltd. All rights reserved.

Soil Conditioning and Ground Monitoring for Shield Tunnelling

Soil conditioning consists of mixing and remolding the natural material during the mechanical excavation of tunnels, generally at low depth, with additives, in order to obtain suitable properties of plasticity and consistency for the excavated material, so becoming able to apply a counterpressure against natural earth pressure and groundwater flow towards the excavation chamber. The assessment and the control of the soil parameters and of machine performance are fundamental for a regular and safe excavation, also with regards to surface stability. This paper mainly focus on testing approach aimed to the proper soil conditioning with EPB shields, whose results have been validated at real scale. The influence of the water content and the amount of conditioning foam has been studied by the Authors. A proper definition of conditioning parameters can allow to extend the application field of Earth Pressure Balance (EPB) tunnel machines to various grain soil distribution, even in weak rock formations (e.g. siltstone or flysch). Importance of conditioning is reflected also on the possibility of a proper spoil disposal or better for its reuse.

Mal (MyD88-adapter-like) is required for Toll-like receptor-4 signal transduction

The recognition of microbial pathogens by the innate immune system involves Toll-like receptors (TLRs), which recognize pathogen-associated molecular patterns. Different TLRs recognize different pathogen-associated molecular patterns, with TLR-4 mediating the response to lipopolysaccharide from Gram-negative bacteria. All TLRs have a Toll/IL-1 receptor (TIR) domain, which is responsible for signal transduction. MyD88 is one such protein that contains a TIR domain. It acts as an adapter, being involved in TLR-2, TLR-4 and TLR-9 signalling; however, our understanding of how TLR-4 signals is incomplete. Here we describe a protein, Mal (MyD88-adapter-like), which joins MyD88 as a cytoplasmic TIR-domain-containing protein in the human genome. Mal activates NF-kappaB, Jun amino-terminal kinase and extracellular signal-regulated kinase-1 and -2. Mal can form homodimers and can also form heterodimers with MyD88. Activation of NF-kappaB by Mal requires IRAK-2, but not IRAK, whereas MyD88 requires both IRAKs. Mal associates with IRAK-2 by means of its TIR domain. A dominant negative form of Mal inhibits NF-kappaB, which is activated by TLR-4 or lipopolysaccharide, but it does not inhibit NF-kappaB activation by IL-1RI or IL-18R. Mal associates with TLR-4. Mal is therefore an adapter in TLR-4 signal transduction.