Signal Height Research Articles

Immobilization of metallothionein to carbon paste electrode surface via anti-MT antibodies and its use for biosensing of silver

In this paper, heavy metal biosensor based on immobilization of metallothionein (MT) to the surface of carbon paste electrode (CPE) via anti-MT-antibodies is reported. First, the evaluation of MT electroactivity was done. The attention was focused on the capturing of MT to the CPE surface. Antibodies incorporated and mixed into carbon paste were stable; even after two weeks the observed changes in signal height were lower than 5%. Further, the interaction of MT with polyclonal chicken antibodies incorporated in carbon paste electrode was determined by square-wave voltammetry. In the voltammogram, two signals – labelled as cys MT and W a – were observed. The cys MT corresponded to –SH moieties of MT and W a corresponded to tryptophan residues of chicken antibodies. Time of interaction (300 s) and MT concentration (125 μg/ml) were optimized to suggest a silver(I) ions biosensor. Biosensor (CPE modified with anti-MT antibody) prepared under the optimized conditions was then used for silver(I) ions detection. The detection limit (3 S/ N) for silver(I) ions was estimated as 0.5 nM. The proposed biosensor was tested by detection spiking of silver(I) ions in various water samples (from very pure distilled water to rainwater). Recoveries varied from 74 to 104%.

Heparin sodium compliance to the new proposed USP monograph: Elucidation of a minor structural modification responsible for a process dependent 2.10 ppm NMR signal

Heparin is a highly sulfated hetero polysaccharide mixture found and extracted from mammalian tissues. It has been widely used as an anticoagulant drug during the past decades. In the new proposed USP heparin monograph, the 1H NMR acceptance criteria to prevent contamination by over sulfated chondroitin sulfate (OSCS), or other persulfated glycosaminoglycans, specifies that no unidentified signals greater than 4% of the mean of signal height of 1 and 2 [1] should be present in the following ranges: 0.10–2.00, 2.10–3.20, and 5.70–8.00 ppm. However, those criteria do not take into account the impact of potential structural modifications generated by the heparin manufacturing processes. In fact, starting from pig mucosa, heparin purification involves oxidizing reagents such as sodium peroxide, potassium permanganate and peracetic acid. In the present work, we demonstrate that potassium permanganate treated heparins show a small but characteristic extra signal at 2.10 ppm. Controlled heparinase I depolymerisation is used to target and excise the oligosaccharide responsible for this extra signal from the polysaccharide backbone. By using orthogonal chromatographic techniques, the fingerprint oligosaccharide was isolated and its structure elucidated. Without the identification of this structural moiety, such purified heparins may have been considered as non-compliant drug substance and not suitable for pharmaceutical use.

Capacitance-voltage spectroscopy on InAs quantum dot valence band states in tilted magnetic fields

We have performed capacitance-voltage (C-V) spectroscopy on the valence band states of InAs quantum dots (QDs) in magnetic fields tilted with respect to the sample plane. We observe an unexpected behaviour for the height of the capacitance signal as function of the in-plane magnetic field, when varying the perpendicular field component, for the first two charging peaks corresponding both to an s-like ground state. With increasing perpendicular field component, the signal height for the first peak decreases strongly and for the second peak the maximum signal height is observed at finite in-plane field values. These observations are not in agreement with the usual assumption that the height-vs.-in-plane-field traces reflect the in-plane k-space probability distribution of the charged QD level because an s-like state should not vary its general shape with perpendicular field. The qualitatively different behaviour of the two peaks is attributed tentatively to correlation effects when the second electron is charged.

Neutron-gamma discrimination by pulse analysis with superheated drop detector

Superheated drop detector (SDD) consisting of drops of superheated liquid of halocarbon is irradiated to neutrons and gamma-rays from 252Cf fission neutron source and 137Cs gamma source separately. The analysis of pulse height of the signals in the neutron and gamma-ray sensitive temperature provides strong information on the identification of neutron and gamma-ray induced events.

Highly selective ionic liquid-based microextraction method for sensitive trace cobalt determination in environmental and biological samples

A simple and rapid dispersive liquid–liquid microextraction procedure based on an ionic liquid (IL-DLLME) was developed for selective determination of cobalt (Co) with electrothermal atomic absorption spectrometry (ETAAS) detection. Cobalt was initially complexed with 1-nitroso-2-naphtol (1N2N) reagent at pH 4.0. The IL-DLLME procedure was then performed by using a few microliters of the room temperature ionic liquid (RTIL) 1-hexyl-3-methylimidazolium hexafluorophosphate [C 6mim][PF 6] as extractant while methanol was the dispersant solvent. After microextraction procedure, the Co-enriched RTIL phase was solubilized in methanol and directly injected into the graphite furnace. The effect of several variables on Co–1N2N complex formation, extraction with the dispersed RTIL phase, and analyte detection with ETAAS, was carefully studied in this work. An enrichment factor of 120 was obtained with only 6 mL of sample solution and under optimal experimental conditions. The resultant limit of detection (LOD) was 3.8 ng L −1, while the relative standard deviation (RSD) was 3.4% (at 1 μg L −1 Co level and n = 10), calculated from the peak height of absorbance signals. The accuracy of the proposed methodology was tested by analysis of a certified reference material. The method was successfully applied for the determination of Co in environmental and biological samples.

Sensing of CO 2 at room temperature using work function readout of (hetero-)polysiloxanes sensing layers

Organically modified silicates based on primary amino groups are known to be CO 2 sensitive, as they can undergo a reversible acid base reaction. In order to generate detectable CO 2 signals and to limit the cross-sensitivity to humidity, some sources suggest that these materials should be operated at higher temperatures (50–70 °C). In this paper, a new variant of CO 2 sensing is to be presented, namely a combination of work function readout and organically modified silicates, which yields CO 2 detection even at room temperature. Kelvin probe measurements are used for work function readout. The layers are intended to be used in a “Floating Gate Field Effect Transistors” (FGFETs) sensing platform (mySens) by Micronas. The reversible interaction of CO 2 with spin-coated heteropolysiloxane sensitive layers results in changes of the work function with typical signal heights of 15–20 mV (change from 400 to 4000 ppm CO 2) and response times of only a few minutes. Also, results will be presented regarding variations in the chemical nature of the films. The findings summarized in this paper point towards the possibility of a new room temperature CO 2 sensor, which comprises fast response times and sufficient sensitivity for ambient CO 2 variations.

Complexes of glutathione with heavy metal ions as a new biochemical marker of aquatic environment pollution

Reduced glutathione (GSH) plays a number of key roles in many biochemical pathways. This peptide is highly reactive and forms conjugates with other molecules via its sulfhydryl moiety. The interactions of the common heavy metal pollutant Cd(II) with GSH were determined by using the Brdicka reaction to evaluate whether this technique would be suitable as a biomarker. After GSH interaction with Cd(II) ions, two characteristic changes in the measured voltammogram were observed: Cat2 signal height decreased, and a new signal called P1 was found. The observed signal probably relates to the formation of a GSH-heavy metal ion complex adsorbed on the surface of the working electrode. When the interaction of GSH with cisplatin was studied, the same characteristic changes in the voltammogram were observed, which confirmed our hypothesis. Moreover, changes in the height of P1 and Cat2 signals with increasing time of GSH interaction with Cd(II) ions and/or cisplatin were also investigated. Cat2 peak height decreased proportionally with increasing time of interaction. This decrease can be explained by shielding of free sulfhydryl moiety by heavy metal ions, so it cannot catalyze the evolution of hydrogen from the supporting electrolyte. In addition, we found that, with increasing time of the interaction, the P1 signal was enhanced and shifted to more positive potentials for both Cd(II) ions and cisplatin.

Determination of serum propofol concentrations by breath analysis using ion mobility spectrometry

We aimed to measure propofol concentrations in exhaled air with an ion mobility spectrometer coupled to a multicapillary column for pre-separation (MCC-IMS). In addition, we aimed to compare the values of these measurements with serum propofol concentrations, as determined by gas chromatography-mass spectrometry (GC-MS). Thirteen patients, ASA I or II, undergoing elective ENT surgery were studied. Anaesthesia was induced with propofol 2.1 (0.7) mg kg(-1), rocuronium 0.5 (0.1) mg kg(-1), and remifentanil 0.5 microg kg(-1) min(-1). After tracheal intubation, anaesthesia was maintained with a continuous infusion of propofol 3.9 (1.8) mg kg(-1) h(-1) and remifentanil 0.5 microg kg(-1) min(-1). Simultaneously, a venous blood sample was obtained. Propofol concentrations in serum were determined by GC-MS and compared with the height of the respective propofol signals achieved by MCC-IMS. Twenty-four pairs of samples were obtained. The comparison of propofol concentrations in exhaled air and serum presented a bias of -10.5% and a precision of +/- 12.3%. With these values, the 95% limits of agreement were 14.1% and -35.1%. MCC-IMS may be a suitable method to determine propofol concentrations in exhaled air, and may be used to predict propofol concentrations in serum.

Comparison of Metallothionein Detection by Using Brdicka Reaction and Enzyme‐Linked Immunosorbent Assay Employing Chicken Yolk Antibodies

AbstractIn this study, two methods for metallothioneins (MT) determination in a biological sample were compared. Particularly, twenty five human and nine pig blood serum samples and liver and kidney samples from thirty five carps (Cyprinus carpio) were analyzed by enzyme‐linked immunosorbent assay and differential pulse voltammetry Brdicka's reaction. The results obtained by these two methods were in good agreement. For commercially available MT standard the correlation coefficient between the single concentrations signal height was 0.99. In biological samples the correlation coefficients were 0.90 for fish liver and kidney samples, 0.91 for pig blood serum and 0.93 for human blood serum.

Detection of CO2 with (Hetero-) Polysiloxanes sensing layers by the change of work function at room temperature

Zhou et al. [1] and Endres et al. [2] reported on CO2-sensing using primary amino groups of Heteropolysiloxanes at higher temperatures (50-70°C) by means of a mass sensitive and a capacitive readout. Based on this work the read out of the change in the work function is investigated. The reversible interaction of CO2 with spin-coated Heteropolysiloxanes sensitive layers results in changes of the work function with typical signal heights of 15-20 mV (from 400 ppm up to 4000 ppm). Results are presented regarding variations in the chemical nature of the films. This opens up the possibility for a new ambient temperature CO2-sensor which shows fast response times as well as a high sensitivity for concentrations exceeding 400 ppm CO2 (background in atmosphere).

Influence of intermediate hydrogel layer and amount of binding sites on the signal response of surface acoustic wave biosensors

Surface acoustic wave (SAW) devices based on horizontally polarized surface shear waves allow direct and label-free detection of biomolecules in real time. Binding reactions on SAW sensor surfaces are detected by determining changes in surface wave velocity caused mainly by mass loading or viscoelasticity changes in the sensing layer. In many cases, analyte-specific capture molecules or ligands are covalently bound on the sensor surface via an intermediate hydrogel layer. It is a fact that the SAW signal response strongly depends on the nature of the hydrogel matrix due to different relative changes of its acousto-mechanical parameters such as density and viscoelasticity. We studied the effect of different hydrogels in two affinity assays. One assay used a low amount of immobilized capture molecules, the other a high amount of immobilized ligands as binding sites in the sensing layer. Significant variations of the SAW signal response were observed. However, performing the assay with immobilized capture molecules resulted in decreasing signal height with increasing molar mass of the hydrogel whereas a reverse signal behavior was obtained performing the assay with immobilized ligands. This means that each affinity system requires its specific hydrogel matrix to obtain maximal signal response.

Signal height in silicon pixel detectors irradiated with pions and protons

Pixel detectors are used in the innermost part of multi-purpose experiments at the Large Hadron Collider (LHC) and are therefore exposed to the highest fluences of ionising radiation, which in this part of the detectors consists mainly of charged pions. The radiation hardness of the detectors has been tested thoroughly up to the fluences expected at the LHC. In case of an LHC upgrade the fluence will be much higher and it is not yet clear up to which radii the present pixel technology can be used. To establish such a limit, pixel sensors of the size of one CMS pixel readout chip (PSI46V2.1) have been bump bonded and irradiated with positive pions up to 6 × 10 14 n eq / cm 2 at PSI and with protons up to 5 × 10 15 n eq / cm 2 . The sensors were taken from production wafers of the CMS barrel pixel detector. They use n-type DOFZ material with a resistance of about 3.7 k Ω cm and an n-side read out. As the performance of silicon sensors is limited by trapping, the response to a Sr-90 source was investigated. The highly energetic beta-particles represent a good approximation to minimum ionising particles. The bias dependence of the signal for a wide range of fluences will be presented.

Breath analysis—performance and potential of ion mobility spectrometry

Ion mobility spectrometry is a fast and sensitive analytical method for the detection of gas phase analytes in the ppbv–pptv range under ambient conditions (pressure and temperature). Ion mobility spectrometers coupled with rapid pre-separation like multi-capillary columns (MCC/IMS) are suitable for the selective characterization of complex and humid mixtures. Recently, MCC/IMS have been applied to analyses of human breath for early diagnosis as well as medication and therapy control. The complete procedure of breath analyses including evaluation and interpretation of the data obtained is demonstrated for the first time on exhaled breath after the consumption of a particular candy as an example. An MCC/IMS equipped with a β-radiation source (63Ni) requires 5 to 10 min for a complete analysis of exhaled breath. Retention time and reduced ion mobility of the detected signals are compared to an analyte database for the identification of the related analytes. These findings were successfully validated by gas-chromatographic mass spectroscopy of the headspace of the candy via solid-phase micro-extraction and of breath samples on Tenax adsorption tubes. Furthermore, signal height of particular analyte signals as a measure for their concentration was used to monitor the concentration development with time. This exemplary investigation demonstrates that MCC/IMS is a powerful and rapid non-invasive tool for human breath analyses. The method can be used for medical applications (diagnosis, therapy control, metabolic profiling) as well as for a general determination of the metabolic state of a subject (medication, nutrition, fasting). The demonstrated procedure is independent of whether the analytes detected in breath are caused by nutrition or medication or whether they are metabolite characteristics for a particular disease. Therefore, it can directly be transferred to any relevant peak pattern.

Characteristics and natural course of vertebral endplate signal (Modic) changes in the Danish general population

BackgroundVertebral endplate signal changes (VESC) are more common among patients with low back pain (LBP) and/or sciatica than in people who are not seeking care for back pain. The distribution and characteristics of VESC have been described in people from clinical and non-clinical populations. However, while the clinical course of VESC has been studied in patients, the natural course in the general population has not been reported. The objectives of this prospective observational study were to describe: 1) the distribution and characteristics of VESC in the lumbar spine, 2) its association with disc degeneration, and 3) its natural course from 40 to 44 years of age.MethodsThree-hundred-and-forty-four individuals (161 men and 183 women) sampled from the Danish general population had MRI at the age of 40 and again at the age of 44. The following MRI findings were evaluated using standardised evaluation protocols: type, location, and size of VESC, disc signal, and disc height. Characteristics and distribution of VESC were analysed by frequency tables. The association between VESC and disc degeneration was analysed by logistic regression analysis. The change in type and size of VESC was analysed by cross-tabulations of variables obtained at age 40 and 44 and tested using McNemar's test of symmetry.ResultsTwo-thirds (67%) of VESC found in this study were located in the lower part of the spine (L4-S1). VESC located at disc levels L1-L3 were generally small and located only in the anterior part of the vertebra, whereas those located at disc levels L4-S1 were more likely to extend further into the vertebra and along the endplate. Moreover, the more the VESC extended into the vertebra, the more likely it was that the adjacent disc was degenerated. The prevalence of endplate levels with VESC increased significantly from 6% to 9% from age 40 to 44. Again, VESC that was only observed in the endplate was more likely to come and go over the four-year period compared with those which extended further into the vertebra, where it generally persisted.ConclusionThe prevalence of VESC increased significantly over the four-year period. Furthermore, the results from this study indicate that the distribution of VESC, its association with disc degeneration and its natural course, is dependent on the size of the signal changes.

A critical appraisal of factors affecting the accuracy of results obtained when using flow cytometry in stem cell investigations: Where do you put your gates?

Flow cytometry is used extensively in stem cell investigations but there is wide variation in the methods used for data analysis between laboratories. Data analysis can be challenging in stem cell biology as there is often no clear distinction between positive and negative populations. We have undertaken a critical appraisal of factors that affect the accuracy of results in stem cell applications. We used mouse embryonic stem cells and determined the expression of three common antigens in stem cell investigations, namely CD15, CD184, and c-kit. We have compared different cell preparation methods and gating strategies and also evaluated the use of isotype controls and unstained cells as controls for the identification of positive populations. The use of a "doublet discriminator" using a side scatter area signal versus side scatter height signal dot plot to identify single cells for analysis increases the accuracy of results regardless of the method used to dissociate cells. Isotype controls can be helpful in mimicking cellular nonspecific binding of the experimental antibody reaction. Isotype controls behave differently on stem cells at different stages of differentiation. Analysis of a viable single cell population with careful selection of control cell populations increases the accuracy of results.

Effects of electrode microstructures of superconducting tunnel junctions on X-ray spectroscopy

Superconducting tunnel junction (STJ) detectors are promising for energy-dispersive X-ray spectroscopy. We are developing array detectors with a sensitive area of 4 mm 2, which consist of 100 of Nb/Al/AlO x /Al/Nb STJs having a size of 200 × 200 μm 2 in order to obtain a good statistic for a short measurement time while keeping an excellent energy resolution for element selection in X-ray material analysis. In the course of this development, we analyzed the microstructures of the layers of Nb, Nb/Al, Nb/Al/AlO x /Al and Nb/Al/AlO x /Al/Nb deposited on Si substrates by using AFM (Atomic Force Microscope) and cross-sectional TEM (Transmission Electron Microscope). We found that the grain sizes of the polycrystalline films and the interface boundaries in the top electrodes are quite different from those in the bottom electrodes in a nanometer scale. It is expected that these different microstructures cause different heights of the pulse signals for the X-ray photon absorption events in the top and bottom electrodes.

Beam–plasma interaction experiments using electromagnetically driven shock waves

Electromagnetically driven shock tubes compatible with in-beam experiments have been developed to examine the stopping power of hot or warm matter over a wide temperature range. The beam–plasma coupling constant γ was calculated under various operating conditions of the shock tube. We found that γ∼0.1 is achievable with 10 keV/u Pb ions and a fully ionized plasma produced by a shock wave with 70–80 km/s in a hydrogen gas of 6–9 kPa. The dissociation effect on the hydrogen stopping power for low-energy protons was also evaluated and a 40–50% increase in the stopping cross-section of dissociated hydrogen was predicted in a projectile energy region of 10–40 keV. For the demonstration of the energy loss measurement using shock-heated gas targets, the developed shock tube was embedded into the beam line and tested on its shock-production abilities. In the preliminary experiment using 375 keV/u carbon projectiles, we successfully detected the signal of a carbon ion penetrating a shock-heat hydrogen target and observed a decrease in the signal height, which probably corresponds to the energy loss.

Influence of Growth Rate and C/Si-Ratio on the Formation of Point and Extended Defects in 4H-SiC Homoepitaxial Layers Investigated by DLTS

4H-SiC epilayers are homoepitaxially grown on 4H-SiC substrates with different C/Si-ratios and different growth rates by the chemical vapour deposition method. DLTS investigations are applied in order to trace energetically deep states of electrically active point defects and extended defects, which may act as the source for the degradation of electronic devices. In addition, the dependence of the DLTS signal heights on the filling pulse length is studied.

Electrochemical Investigation of Strontium–Metallothionein Interactions – Analysis of Serum and Urine of Patients with Osteoporosis

AbstractThe main aim of this paper is to study interaction between strontium and metallothionein (MT), and to determine changes in strontium and thiols (MT, reduced glutathione, cysteine, and homocysteine) level in plasma, serum, and urine samples of patients treated with strontium ranelate (SrR). To investigate the interactions between MT and strontium(II) ions, adsorptive transfer stripping technique coupled with differential pulse voltammetry (DPV) the Brdicka reaction was employed. Besides standard Brdicka signals (Co, RS2Co, Cat1, Cat2, Cat3), we observed new signal related to Sr‐MT interaction. Further we investigated the effect of various time of interaction, concentration of strontium(II) ions and temperature of supporting electrolyte on Brdicka signals. Optimal time of interaction was 240 s. Under temperature of supporting electrolyte 20 °C, we measured linear dependence of Cat3 signal height on strontium(II) ions concentration. After that we have investigated the possibility of strontium‐MT interactions, we were interested in strontium, MT and low molecular mass thiols levels in serum and urine of patients treated with strontium(II) ions to cure osteoporosis. Strontium concentration determined by atomic absorption spectrometry was 55±5 μg/L before and 10,500±1,400 μg/L at the 30th day of SrR administration. Levels of metallothionein in serum ranged from 0.1 to 6.4 μM. Correlation between serum strontium concentration and MT level was determined and correlation coefficient was R=0.93.

Regulation of Influenza Virus-Caused Oxidative Stress by Kegan Liyan Oral Prescription, as Monitored by Ascorbyl Radical ESR Signals

To study the oxidative stress level of the influenza virus A FM1 subset-infected mouse in intranasal inhalation as a model, we employ an ascorbyl radical's ESR (electron spin resonance) spectrum as an oxidative stress biomarker. These infected mice were pretreated with Ribavirin, ascorbic acid, superoxide dismutase (SOD) or Kegan Liyan oral prescription (KGLY, proprietary Chinese medicine for influenza and common cold) in the stomach tube for 3 days, and then followed by the virus-infecting for 4 days. On the 4th day, samples were collected. It is recognized the strength of ascorbyl radical's ESR signal (A(-.)) (a(H4 = 0.177) Gauss, g = 2.00517) denotes oxidative stress level in vivo and in vitro. The magnitude of ESR spectrum (28.65 +/- 10.71 AU) in mice infected with influenza virus was significantly higher than those of healthy control mice (19.10 +/- 3.61 AU). Serum A(-.) in mice treated with Ribavirin, ascorbic acid, SOD and KGLY declined to 19.70 +/- 6.05, 18.50 +/- 2.93 and 16.25 +/- 3.59, 18.40 +/- 2.14 AU respectively. It is close to A(-.) signal height in healthy controls via down-regulation of the influenza virus-caused oxidative stress level getting decline in the lung index of pneumonia as compare to those of untreated healthy and the influenza virus infected mice pneumonia. It is well known that SOD can prevent the influenza virus pneumonia enhancing mouse survival rate; Ribavirin can treat viral diseases. Data from this study suggested that KGLY may indirectly relieve influenza virus-infected pneumonia via down- regulation of virus caused oxidative stress coupled with a redox reaction cascade as ribavirin, ascorbic acid and SOD.