Fast Peak Research Articles

Dynamic high‐resolution computer simulation of electrophoretic enantiomer separations with neutral cyclodextrins as chiral selectors

GENTRANS, a comprehensive one-dimensional dynamic simulator for electrophoretic separations and transport, was extended for handling electrokinetic chiral separations with a neutral ligand. The code can be employed to study the 1:1 interaction of monovalent weak and strong acids and bases with a single monovalent weak or strong acid or base additive, including a neutral cyclodextrin, under real experimental conditions. It is a tool to investigate the dynamics of chiral separations and to provide insight into the buffer systems used in chiral capillary zone electrophoresis (CZE) and chiral isotachophoresis. Analyte stacking across conductivity and buffer additive gradients, changes of additive concentration, buffer component concentration, pH, and conductivity across migrating sample zones and peaks, and the formation and migration of system peaks can thereby be investigated in a hitherto inaccessible way. For model systems with charged weak bases and neutral modified β-cyclodextrins at acidic pH, for which complexation constants, ionic mobilities, and mobilities of selector-analyte complexes have been determined by CZE, simulated and experimentally determined electropherograms and isotachopherograms are shown to be in good agreement. Simulation data reveal that CZE separations of cationic enantiomers performed in phosphate buffers at low pH occur behind a fast cationic migrating system peak that has a small impact on the buffer composition under which enantiomeric separation takes place.

Non-linear Modelling of Fasting Plasma Glucose Concentration and Peak 2-hour Insulin Response to Glucose Challenge in Australian Aboriginal People

Introduction The current American Diabetes Association recommended diagnostic criterion for type 2 diabetes mellitus of fasting plasma glucose (FPG) ≥7.0 mmol/l is based on the point at which risk for macrovascular and microvascular complications substantially increases [1,2]. The WHO continues to recommend assessment of plasma glucose level by oral glucose tolerance

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Diazoxide

Effect of Cholesterol on the Pore Dilation of TRPV1

The transient receptor potential vanilloid 1 (TRPV1) ion channel is expressed in nociceptors, where pharmacological modulation of its function may offer a way of easing pain and neurogenic inflammation processes in the human body. The aim of this study was to investigate how depletion of cholesterol from the cell membrane may modulate ion-permeability of the TRPV1 ion channel expressed in CHO-cells. The ion-permeability properties of TRPV1 measured with whole-cell patch clamp on a Chinese hamster ovary (CHO) cell line expressing TRPV1. Sustained capsaicin-induced activation of TRPV1 with N-methyl-D-glucamine (NMDG) as the sole external cation, generated a biphasic current, with a first outward current and a second inward current. Similarly, sustained proton-activation (pH 5.5) of TRPV1 in the absence of external calcium also generated a biphasic current, with a first fast current peak followed by a second larger one. Also, patch clamp recordings of reversal potentials revealed a change of the ion-permeability during prolonged activation of the TRPV1 channel in extremely low extracellular calcium. Our findings show that depletion of cholesterol from the cell membrane inhibited both the second current resulting from sustained agonist-activation, and the change in ion-permeability of the TRPV1 channel. Our results propose a novel mechanism by which cholesterol-depletion may modulate the ion channel function of TRPV1, which may constitute a novel pharmacological based approach for the treatment of pain and neurogenic pain.

Semi-Explicit Modelling of Watersheds with Urban Drainage Systems

In rainfall-runoff modelling of urbanized and semi-urbanized watersheds, the urban drainage systems considerably influence runoff propagation time. In small scale watersheds, the drainage network may be modelled explicitly. In contrast, for larger watersheds, most hydrological models are based on a rough representation of the effects of drainage systems, thus failing to represent the rapidly-varying real flow dynamics.Therefore, a trade-off methodology has been developed to account for impervious surfaces and drainage effects accurately, without the need for modelling the entire drainage network in detail. Undrained impervious areas have been distinguished from drained ones. Rain falling on the former has been discharged as overland flow, whereas flow on the later has been routed separately using “virtual pipes”, which enable a simplified process-oriented modelling of the drainage network.The methodology has been applied to a 130 km2 Belgian catchment, resulting in the simulation of fast flow peaks, which do not appear when the effect of the drainage network is neglected.

Highly sensitive and selective analysis of urinary steroids by comprehensive two-dimensional gas chromatography combined with positive chemical ionization quadrupole mass spectrometry.

Comprehensive two dimensional gas chromatography (GC × GC) provides greater separation space than conventional GC. Because of fast peak elution, a time of flight mass spectrometer (TOFMS) is the usual structure-specific detector of choice. The quantitative capabilities of a novel GC × GC fast quadrupole MS were investigated with electron ionization (EI), and CH(4) or NH(3) positive chemical ionization (PCI) for analysis of endogenous urinary steroids targeted in anti-doping tests. Average precisions for steroid quantitative analysis from replicate urine extractions were 6% (RSD) for EI and 8% for PCI-NH(3). The average limits of detection (LODs) calculated by quantification ions for 12 target steroids spiked into steroid-free urine matrix (SFUM) were 2.6 ng mL(-1) for EI, 1.3 ng mL(-1) for PCI-CH(4), and 0.3 ng mL(-1) for PCI-NH(3), all in mass scanning mode. The measured limits of quantification (LOQs) with full mass scan GC × GC-qMS were comparable with the LOQ values measured by one-dimensional GC-MS in selected ion monitoring (SIM) mode. PCI-NH(3) yields fewer fragments and greater (pseudo)molecular ion abundances than EI or PCI-CH(4). These data show that a benchtop GC × GC-qMS system has the sensitivity, specificity, and resolution to analyze urinary steroids at normal urine concentrations, and that PCI-NH(3), not currently available on most GC × GC-TOFMS instruments, is of particular value for generation of structure-specific ions.

Relationship between neonatal screening results by HPLC and the number of α-thalassaemia gene mutations; consequences for the cut-off value

To evaluate the relationship between FAST peak percentage by adapted Bio-Rad Vnbs analysis using the valley-to-valley integration and genotypes with the aim to improve differentiation between severe α-thalassaemia forms (HbH disease) and the milder disease types. DNA analysis for α-thalassaemia was performed on 91 dried blood spot samples presenting normal and elevated FAST peak levels, selected during three years of Dutch national newborn screening. Significant differences were found between samples with and without α-thalassaemia mutations, regardless of the genetic profiles. No significant difference was demonstrated between HPLC in -α/αα and -α/-α, between -α/-α and - -/αα and between - -/αα and - -/-α genotypes. This study confirms that the percentage HbBart's, as depicted by the FAST peak, is only a relative indication for the number of α genes affected in α-thalassaemia. Based on the data obtained using the modified Bio-Rad Vnbs software, we adopted a cut-off value of 22.5% to discriminate between possible severe α-thalassaemia or HbH disease and other α-thalassaemia phenotypes. Retrospectively, if this cut-off value was utilized during this initial three-year period of neonatal screening, the positive predictive value would have been 0.030 instead of 0.014.

Sleep EEG fingerprints reveal accelerated thalamocortical oscillatory dynamics in Williams syndrome

Sleep EEG alterations are emerging features of several developmental disabilities, but detailed quantitative EEG data on the sleep phenotype of patients with Williams syndrome (WS, 7q11.23 microdeletion) is still lacking. Based on laboratory (Study I) and home sleep records (Study II) here we report WS-related features of the patterns of antero-posterior 8-16 Hz non-rapid-eye-movement (NREM) sleep EEG power distributions. Participants in Study I were 9 WS and 9 typically developing (TD) controls matched for age (14-29 years) and sex, and sleeping for two consecutive nights in the laboratory. WS participants were characterized by region-independent decreases in 10.50-12.50 Hz and central increases in 14.75-15.75 Hz EEG power. Region-independent decreases and increases in z-scores of the spectra were observed in the 10.25-12.25 Hz and 14-16 Hz ranges, respectively. Moreover, in the EEG spectra of participants with WS a lower probability for the emergence of a frontally dominant peak was observed. Parietal fast sigma peaks and the antero-posterior shifts in power distributions were of higher frequencies in WS (~1 Hz difference). A 1 year follow-up of 9 WS and 3 TD participants, as well as their inclusion into larger samples (20 WS and 20 TD, age: 6-29 years) of a two-night ambulatory home polysomnography study confirmed the WS-specific decrease in alpha/low sigma power (8-11.75 Hz) and the pattern of z-score differences (decreases: 8.50-11.25 Hz; increases: 13.5-14 Hz), including the antero-posterior shifts in power distribution (0.5 Hz) and some features of the spectral peaks. Altogether these data suggest a decrease in alpha/low sigma power, as well as a redistribution of NREM sleep 8-16 Hz EEG power toward the higher frequencies and/or a higher frequency of NREM sleep thalamocortical oscillations in WS.

Novel Fast Peak Detector for Single- or Three-phase Unsymmetrical Voltage Sags

In the present paper, a novel fast peak detector for single- or three-phase unsymmetrical voltage sags is proposed. The proposed detector is modified from a single-phase digital phase-locked loop based on a d-q transformation using an all-pass filter (APF). APF generates a virtual phase with <TEX>$90^{\circ}$</TEX> phase delay. However, this virtual phase cannot reflect a sudden change of the grid voltage in the moment of voltage sag, which causes a peak value to be significantly distorted and to settle down slowly. Specifically, the settling time of the peak value is too long when voltage sag occurs around a zero crossing, such as phase <TEX>$0^{\circ}$</TEX> and <TEX>$180^{\circ}$</TEX>. This paper describes the operating principle of the APF problem and proposes a modified all-pass filter (MAPF) to mitigate the inherent APF problem. In addition, a new fast peak detector using MAPF is proposed. The proposed detector is able to calculate a peak value within 0.5 ms, even when voltage sag occurs around zero crossing. The proposed fast peak detector is compared with the conventional detector using APF. Results show that the proposed detector has faster detection time in the whole phase range. Furthermore, the proposed fast peak detector can be effectively applied to unsymmetrical three-phase voltage sags. Simulation and experimental results verify the advantages of the proposed detector and MAPF.

Slow protein evolutionary rates are dictated by surface–core association

Why do certain proteins evolve much slower than others? We compared not only rates per protein, but also rates per position within individual proteins. For ∼90% of proteins, the distribution of positional rates exhibits three peaks: a peak of slow evolving residues, with average log(2)[normalized rate], log(2)μ, of ca. -2, corresponding primarily to core residues; a peak of fast evolving residues (log(2)μ ∼ 0.5) largely corresponding to surface residues; and a very fast peak (log(2)μ ∼ 2) associated with disordered segments. However, a unique fraction of proteins that evolve very slowly exhibit not only a negligible fast peak, but also a peak with a log(2)μ ∼ -4, rather than the standard core peak of -2. Thus, a "freeze" of a protein's surface seems to stop core evolution as well. We also observed a much higher fraction of substitutions in potentially interacting residues than expected by chance, including substitutions in pairs of contacting surface-core residues. Overall, the data suggest that accumulation of surface substitutions enables the acceptance of substitutions in core positions. The underlying reason for slow evolution might therefore be a highly constrained surface due to protein-protein interactions or the need to prevent misfolding or aggregation. If the surface is inaccessible to substitutions, so becomes the core, thus resulting in very slow overall rates.

Effects of dietary carbohydrate sources on plasma glucose, insulin and IGF-I levels in multiparous sows

Effects of different carbohydrate sources on plasma glucose, insulin and insulin-like growth factor-I (IGF-I) levels were compared to subsequently be able to study effects of insulin-stimulating diets on follicle development in sows. The following feed components were tested in 12 sows during six consecutive test periods of 9.5 days: dextrose (DEX), sucrose (SUC), lactose (LAC), dextrose plus lactose (DL), sucrose plus lactose (SL), dextrose plus sugarbeet pulp (DSBP) and control (CON). On day 2, 5 and 9 of each test period, plasma glucose (only at day 9), insulin and IGF-I profiles were determined. Despite similar glucose profiles for all diets, the postprandial insulin response was higher for DL and SL compared with CON and LAC; the other diets were intermediate. Plasma IGF-I levels were higher in CON, LAC and SL compared with DSBP, but differences were only marginal. It was concluded that dextrose and sucrose have the potential to stimulate fast and high insulin peaks, especially when combined with additional lactose. Despite the high dextrose in the DSBP diet, the insulin response was flattened, probably due to the viscosity of sugarbeet pulp. The results show that modulation of plasma insulin levels by dietary carbohydrates seems possible in anabolic sows, but IGF-I levels are less easily modified.

Full characterization of laser-accelerated ion beams using Faraday cup, silicon carbide, and single-crystal diamond detectors

Multi-MeV beams of light ions have been produced using the 300 picosecond, kJ-class iodine laser, operating at the Prague Asterix Laser System facility in Prague. Real-time ion diagnostics have been performed by the use of various time-of-flight (TOF) detectors: ion collectors (ICs) with and without absorber thin films, new prototypes of single-crystal diamond and silicon carbide detectors, and an electrostatic ion mass spectrometer (IEA). In order to suppress the long photopeak induced by soft X-rays and to avoid the overlap with the signal from ultrafast particles, the ICs have been shielded with Al foil filters. The application of large-bandgap semiconductor detectors (&amp;gt;3 eV) ensured cutting of the plasma-emitted visible and soft-UV radiation and enhancing the sensitivity to the very fast proton/ion beams. Employing the IEA spectrometer, various ion species and charge states in the expanding laser-plasma have been determined. Processing of the experimental data based on the TOF technique, including estimation of the plasma fast proton maximum and peak energy, ion beam currents and total charge, total number of fast protons, as well as deconvolution processes, ion stopping power, and ion/photon transmission calculations for the different metallic filters used, are reported.

Design and construction of a compact portable pulsed power generator

The low impedance compact pulsed power generators are considered as potential drivers for X-pinch based phase contrast imaging and extreme ultraviolet (EUV) source for lithography. The designed pulsed power generator should provide square-like wave on low impedance load (1–2 Ω) with current amplitude of 100 kA in burst mode. The pulse width of the current wave is 60 ns, and the rise time of the wave should be less than 30 ns. The dimensions of the machine are restricted to be no larger than ∼2 × 1 × 1.5 m, according to the specific potential application of the machine. We adopted a solution based on Marx generator combined with low impedance pulse forming line and V/N type gas switch technology to obtain the required high intensity and fast rise-time current wave. Special efforts have been made to reduce the inductance of V/N switch and low impedance load section, and thus to obtain fast rise time and high peak current, during both physical and mechanical design stages. The high power transient signal diagnostic system, consisting of a capacitive voltage divider and a metal film based current monitor, and the calibration of the diagnostic system are also outlined.

Performance of tourmaline hydrophones at low- and midfrequencies.

Tourmaline hydrophones are well suited for measurement of underwater blasts and other impulsive type sounds having extremely fast rise times and high peak pressures. The physical characteristics of these small, lightweight sensors, however, make them an attractive choice for other applications. To determine their performance at low- and midfrequencies, substitution calibrations of two tourmaline sensors with a calibrated laboratory hydrophone were completed using a J-ll underwater sound transducer in a 100-gal tank. The calibration signals consisted of tone bursts containing 5–20 cycles at frequencies ranging from 250 to 10 000 Hz and having sound pressure levels from 110 to 180 dB re 1 μa. Results indicate that a tourmaline hydrophone coupled to an in-line charge amplifier and preamplifier to provide fixed voltage gains ranging from 100× to 10 000× has excellent performance over this range of frequencies. [Work supported by the National Oceanographic Partnership Program (NOPP) through ONR Award No. N000140710992.]

Loss of olfactory cell adhesion molecule reduces the synchrony of mitral cell activity in olfactory glomeruli

Odours generate activity in olfactory receptor neurons, whose axons contact the dendritic tufts of mitral cells within olfactory bulb glomeruli. These axodendritic synapses are anatomically separated from dendrodendritic synapses within each glomerulus. Mitral cells within a glomerulus show highly synchronized activity as assessed with whole-cell recording from pairs of mitral cells. We examined glomerular activity in mice lacking the olfactory cell adhesion molecule (OCAM). Glomeruli in mice lacking OCAM show a redistribution of synaptic subcompartments, but the total area occupied by axonal inputs was similar to wild-type mice. Stimulation of olfactory nerve bundles showed that excitatory synaptic input to mitral cells as well as dendrodendritic inhibition was unaffected in the knockout. However, correlated spiking in mitral cells was significantly reduced, as was electrical coupling between apical dendrites. To analyse slow network dynamics we induced slow oscillations with a glutamate uptake blocker. Evoked and spontaneous slow oscillations in mitral cells and external tufted cells were broader and had multiple peaks in OCAM knockout mice, indicating that synchrony of slow glomerular activity was also reduced. To assess the degree of shared activity between mitral cells under physiological conditions, we analysed spontaneous sub-threshold voltage oscillations using coherence analysis. Coherent activity was markedly reduced in cells from OCAM knockout mice across a broad range of frequencies consistent with a decrease in tightly time-locked activity. We suggest that synchronous activity within each glomerulus is dependent on segregation of synaptic subcompartments.

Minocycline Suppresses Activation of Nuclear Factor of Activated T Cells 1 (NFAT1) in Human CD4+ T Cells

Minocycline is a tetracycline family antibiotic that has anti-inflammatory and immunomodulatory properties. These properties have shown promise in the treatment of conditions such as rheumatoid arthritis, Huntington disease, and multiple sclerosis. As lymphocyte activation is involved in the pathogenesis of many of these diseases, T cells are postulated to be a primary target in minocycline therapy. Previous studies have demonstrated attenuation of CD4(+) T cell activation by minocycline, but a specific mechanism has not been elucidated. In this study, we investigated the effect of minocycline on the activity of three key transcription factors regulating CD4(+) T cell activation: NF-κB, AP-1 (activator protein 1), and NFAT (nuclear factor of activated T) cells. Our data demonstrate that minocycline selectively impairs NFAT-mediated transcriptional activation, a result of increased phosphorylation and reduced nuclear translocation of the isoform NFAT1. Minocycline increased the activity of the NFAT kinase GSK3 and decreased intracellular Ca(2+) flux, both of which facilitate NFAT1 phosphorylation. These findings provide a novel mechanism for minocycline induced suppression of CD4(+) T cell activation and may better inform the application of minocycline as an immunomodulatory agent.

Contribution of Rho kinase to the early phase of the calcium-contraction coupling in airway smooth muscle

We investigated theoretically and experimentally the role of Rho kinase (RhoK) in Ca(2+)-contraction coupling in rat airways. Isometric contraction was measured on tracheal, extrapulmonary and intrapulmonary bronchial rings. Intracellular [Ca(2+)] was recorded in freshly isolated tracheal myocytes. Stimulation by carbachol (0.3 and 10 μm) and 50 mm external KCl induced a short-time, Hill-shaped contraction obtained within 90 s, followed by a sustained or an additional delayed contraction. Responses of [Ca(2+)](i) to acetylcholine consisted in a fast peak followed by a plateau and, in 42% of the cells, superimposed Ca(2+) oscillations. The RhoK inhibitor Y27632 (10 μm) did not alter the [Ca(2+)](i) response. Whatever the agonist, Y27632 did not modify the basal tension but decreased the amplitude of the short-duration response, without altering the additional delayed contraction. The Myosin Light Chain Phosphatase (MLCP) inhibitor calyculin A increased the basal tension and abolished the effect of RhoK. KN93 (Ca(2+)-calmodulin-dependent protein kinase II inhibitor) and DIDS (inhibitor of Ca(2+)-activated Cl(-) channels) had no influence on the RhoK effect. We built a theoretical model of Ca(2+)-dependent active/inactive RhoK ratio and subsequent RhoK-dependent MLCP inactivation, which was further coupled with a four-state model of the contractile apparatus and Ca(2+)-dependent MLCK activation. The model explains the time course of the short-duration contraction and the role of RhoK by Ca(2+)-dependent activation of MLCK and RhoK, which inactivates MLCP. Oscillatory and non-oscillatory [Ca(2+)](i) responses result in a non-oscillatory contraction, the amplitude of which is encoded by the plateau value and oscillation frequency. In conclusion, Ca(2+)-dependent but CaMK II-independent RhoK activation contributes to the early phase of the contractile response via MLCP inhibition.

The RPC current time structure. Fast current peak measurement in the ATLAS RPC system

We have developed an analog RPC read out based on the observation of the signal produced in the high voltage circuit by many simultaneous particles crossing the same gas gap. The proposed method allows to measure particle densities by far much larger that those accessible to the standard digital read out. We applied this method to study with the ATLAS RPC system the beam “splash” events produced by LHC at the beginning of its activity.

An Automated Data Analysis Pipeline for GC−TOF−MS Metabonomics Studies

Recent technological advances have made it possible to carry out high-throughput metabonomics studies using gas chromatography coupled with time-of-flight mass spectrometry. Large volumes of data are produced from these studies and there is a pressing need for algorithms that can efficiently process and analyze data in a high-throughput fashion as well. We present an Automated Data Analysis Pipeline (ADAP) that has been developed for this purpose. ADAP consists of peak detection, deconvolution, peak alignment, and library search. It allows data to flow seamlessly through the analysis steps without any human intervention and features two novel algorithms in the analysis. Specifically, clustering is successfully applied in deconvolution to resolve coeluting compounds that are very common in complex samples and a two-phase alignment process has been implemented to enhance alignment accuracy. ADAP is written in standard C++ and R and uses parallel computing via Message Passing Interface for fast peak detection and deconvolution. ADAP has been applied to analyze both mixed standards samples and serum samples and identified and quantified metabolites successfully. ADAP is available at http://www.du-lab.org .

Fast scanning cavity offset lock for laser frequency drift stabilization

We have implemented a compact setup for long-term laser frequency stabilization. Light from a stable reference laser and several slave lasers is coupled into a confocal Fabry-Perot resonator. By stabilizing the position of the transmission peaks of the slave lasers relative to successive peaks of the master laser as the length of the cavity is scanned over one free spectral range, the long-term stability of the master laser is transferred to the slave lasers. By using fast analog peak detection and low-latency microcontroller-based digital feedback, with a scanning frequency of 3 kHz, we obtain a feedback bandwidth of 380 Hz and a relative stability of better than 10 kHz at timescales longer than 1 s, a significant improvement on previous scanning-cavity stabilization systems.