Fully automated chemiluminescence immunoassay of insulin using antibody-protein A-bacterial magnetic particle complexes

We report a fully automated sandwich immunoassay for the determination of human insulin using antibody-protein A-bacterial magnetic particle (BMP) complexes and an alkaline phosphatase-conjugated secondary antibody. BMPs bearing protein A-MagA inserted on the external surface of the membrane were prepared in the Magnetospirillum sp. AMB-1 transconjugant for a protein A-magA fusion gene. MagA protein was used as an anchor to attach protein A onto the membrane. Protein A-BMP complexes harvested from transconjugant AMB-1 were subsequently complexed with anti-human insulin antibodies by specific binding between the Z domain of protein A and the Fc component of IgG to form the antibody-protein A-BMP complexes. The complexes were quite monodisperse after the binding of the antibody. The BMPs' monodispersity resulted in high signal and low noise in the immunoassay. The luminescence intensity ((kilocounts/s)/microg of antibody) from antibody-protein A-BMP complexes after immunoreaction was higher than that from BMPs chemically conjugated to an antibody. This was explained by a difference in dispersion. The fully automated sandwich immunoassay system using antibody-protein A-BMP complexes made possible precise assays of human insulin in serum.     

High-throughput nanohole array based system to monitor multiple binding events in real time

We have developed an integrated label-free, real-time sensing system that is able to monitor multiple biomolecular binding events based on the changes in the intensity of extraordinary optical transmission (EOT) through nanohole arrays. The core of the system is a sensing chip containing multiple nanohole arrays embedded within an optically thick gold film, where each array functions as an independent sensor. Each array is a square array containing 10 x 10 nanoholes (150 nm in diameter), occupying a total area of 3.3 mum x 3.3 mum. The integrated system includes a laser light source, a temperature-regulated flow cell encasing the sensing chip, motorized optics, and a charge-coupled detector (CCD) camera. For demonstration purposes, sensing chips containing 25 nanohole arrays were studied for their use in multiplexed detection, although the sensing chip could be easily populated to contain up to 20 164 nanohole arrays within its 0.64 cm2 sensing area. Using this system, we successfully recorded 25 separate binding curves between glutathione S-transferase (GST) and anti-GST simultaneously in real time with good sensitivity. The system responds to binding events in a concentration-dependent manner, showing a sharp linear response to anti-GST at concentrations ranging from 13 to 290 nM. The EOT intensity-based approach also enables the system to monitor multiple bindings simultaneously and continuously, offering a temporal resolution on milliseconds scale that is decided only by the camera speed and exposure time. The small footprint of the sensing arrays combined with the EOT intensity-based approach enables the system to resolve binding events that occurred on nanohole sensing arrays spaced 96 mum apart, with a reasonable prediction of resolving binding events spaced 56 mum apart. This work represents a new direction that implements nanohole arrays and EOT intensity to meet high-throughput, spatial and temporal resolution, and sensitivity requirements in drug discovery and proteomics studies.     

Variations in occupational exposure to magnetic fields among welders in Malaysia

This study attempted to estimate the lifelong magnetic field (MF) exposures of a particular group of welders. Exposure was quantified via measurements, observations and interviews. It was found that these welders face a vast range of lifelong MF exposures depending on the welding processes and duration of the welding tasks performed. This may explain the inconsistency in the results of studies of MF exposures on human health. The mere assessing of the MF exposure levels through spot measurements does not give an overall picture of the total amount of exposure received by the welders as some of these workers performed the welding task throughout the day, whereas others performed this as a part of their job. The exposure to various chemicals in the fume may complicate the interpretation of the elevated health risk among the welders.     

Evaluation of long-term exposure to the magnetic field produced from power lines

For some years, the Italian Regional Agencies for Environmental Protection (ARPA) have been engaged in the evaluation of long-term exposure to 50 Hz magnetic fields for receptors close to power lines. In this study, experiences and procedures are suggested in order to estimate long-term exposure levels in the case of magnetic induction produced contemporaneously by different power lines. In the simplest cases, the influence of a single line is evaluated by verifying the correlation factor between the variation of the current in each line and the measured magnetic induction. When the prevalence of a single line is not clear and systematic, much additional data (voltage, current, phases and geometrical configuration) have to be considered. The case of a school in Livomo near two lines. placed on the same supports, is described and a range of the most probable exposure values is given, corresponding to the phase change of the two current fluxes.     

Iron oxide nanoparticle-based magnetic resonance method to monitor release kinetics from polymeric particles with high resolution

A new method to precisely monitor rapid release kinetics from polymeric particles using super paramagnetic iron oxide nanoparticles, specifically by measuring spin-spin relaxation time (T(2)), is reported. Previously, we have published the formulation of logic gate particles from an acid-sensitive poly-β-aminoester ketal-2 polymer. Here, a series of poly-β-aminoester ketal-2 polymers with varying hydrophobicities were synthesized and used to formulate particles. We attempted to measure fluorescence of released Nile red to determine whether the structural adjustments could finely tune the release kinetics in the range of minutes to hours; however, this standard technique did not differentiate each release rate of our series. Thus, a new method based on encapsulation of iron oxide nanoparticles was developed, which enabled us to resolve the release kinetics of our particles. Moreover, the kinetics matched the relative hydrophobicity order determined by octanol-water partition coefficients. To the best of our knowledge, this method provides the highest resolution of release kinetics to date.     

High-resolution organ dosimetry for human exposure to low-frequencymagnetic fields

Electric fields and current densities induced in an anatomically realistic, high-resolution model of the human body are computed. The results are given for various organs in terms of average and maximum values for 60-Hz uniform magnetic fields in three orthogonal orientations. The effects of variations in the tissue conductivity are evaluated. The data presented can he linearly scaled to frequencies up to 100 kHz without an appreciable error, provided that the changes in tissue conductivity with frequency are taken into account. The scalar potential finite difference (SPFD) method with an appropriate matrix preconditioner and a conjugate gradient solver were used to model the problem. This approach resulted in a high computational efficiency that facilitated high-resolution (3.6 mm voxels) modeling involving 1 736 872 unknowns. All computations were performed on a Hewlett-Packard 9000/735 Unix workstation using under 200 MB of physical memory. Typical computation times were of the order of 25 h. The results show that conductivity variations, source orientation, and model realism can significantly affect the dosimetry values, particularly in comparison with more simplistic models. They also show that the maximum current density in several tissues can exceed 10 mA m^-2 in a 60-Hz, 0.5-mT uniform magnetic field     

A novel automated alternating current biosusceptometry method to characterization of controlled-release magnetic floating tablets of metronidazole

Context: Alternating Current Biosusceptometry is a magnetically method used to characterize drug delivery systems. This work presents a system composed by an automated ACB sensor to acquire magnetic images of floating tablets.

Objective: The purpose of this study was to use an automated Alternating Current Biosusceptometry (ACB) to characterize magnetic floating tablets for controlled drug delivery.

Materials and methods: Floating tablets were prepared with hydroxypropyl methylcellulose (HPMC) as hydrophilic gel material, sodium bicarbonate as gas-generating agent and ferrite as magnetic marker. ACB was used to characterize the floating lag time and the tablet hydration rate, by quantification of the magnetic images to magnetic area. Besides the buoyancy, the floating tablets were evaluated for weight uniformity, hardness, swelling and in vitro drug release.

Results: The optimized tablets were prepared with equal amounts of HPMC and ferrite, and began to float within 4?min, maintaining the flotation during more than 24?h. The data of all physical parameters lied within the pharmacopeial limits. Drug release at 24?h was about 40%.

Conclusions: The ACB results showed that this study provided a new approach for in vitro investigation of controlled-release dosage forms. Moreover, using automated ACB will also be possible to test these parameters in humans allowing to establish an in vitro.in vivo correlation (IVIVC).     

Cellular responses to low dose heavy-ion exposure in human cells

The human lymphoblastoid cell line TK6 was used to study the cellular responses after low-dose (100, 200, 500 mGy) or high-dose (3 Gy) of X rays, C (22 keV.micron-1) and Fe (1000 keV.micron-1) ion exposures. p53 protein induction in individual cells was determined by indirect immunofluorescence staining. Cell-cycle progression after heavy-ion exposure was determined by using a laser scanning cytometer. A characteristic pattern of cell-cycle progression was observed with 3 Gy exposure of Fe ions but not with 100 mGy. Similarly such a pattern with 100 mGy C ion exposure did not match that with 3 Gy. The proportion of p53-induced cells is proportional to the probability of cell being hit by a primary heavy ion. The observed low-dose effect can be reflected in the probability of a hit, although detailed nature about their energy deposition must be considered for more precise estimation of such an effect. New detection methodology must be developed for identification of heavy-ion specific cellular responses.     

Methodology of a study on the French population exposure to 50 Hz magnetic fields

The characterisation of population exposure to a 50-Hz magnetic field (MF) is important for assessing health effects of electromagnetic fields. With the aim of estimating and characterising the exposure of the French population to 50-Hz MFs, two representative samples of the population were made. A random selection method based on the distribution of households in different regions of France was used. The samples were carried out starting from a random polling of telephone numbers of households (listed, unlisted fixed phones and cell phones only). A total of 95,362 telephone numbers were dialed to have 2148 volunteers (1060 children and 1088 adults). They all agreed to carrying an EMDEX II meter, measuring and recording MFs, and to filling out a timetable for a 24-hour period. In this article, the methodology of the sample selection and the collection of all necessary information for the realisation of this study are presented.     

Mass imagineering: Combining human imagination and automated engineering from early education to digital afterlife

Imagineering is a portmanteau combining the words imagination and engineering. The term, imagineering, was coined during the early 1940s. Since the new Millennium, imagineering has grown to the scale of a mass paradigm, because of the increasing scope and availability of automated engineering. Automated engineering is enabled by sophisticated software and artificial intelligence (AI) across the virtual-social-physical convergence (VSP) of consumer devices, desktop machines, and Web platforms. The combining of human imagination with automated engineering is enabling millions of vloggers, self-publishers and makers to imagineer at high speed and low cost. This mass imagineering can range from early education to digital afterlife. It involves makerspaces, fab labs, hackerspaces, as well as use of disparate devices, machines, and platforms “in the wild”. In this paper, three principal contributions to the literature are provided. First, mass imagineering is described from early education to digital afterlife. Second, analysis of structure and agency in mass imagineering is provided. This reveals that individual agency varies in different settings for mass imagineering, but is always more than in mass production and mass customization settings. Third, division of labour analysis of mass imagineering is provided. This reveals that mass imagineering extends labour outside of paid employment, such as in the home, towards greater self-expression and increased entrepreneurship. This is possible because of increasing automated engineering across VSP enabled by devices, machines, and platforms.     

Estimation of human exposure to natural radionuclides using in vivo skull measurements

In a preliminary study, in vivo skull measurements and in vitro urine measurements of 210Pb and nulU have been performed to find out the individual, chronic exposure to waterborne natural radionuclides of a small group of Finnish people. For their domestic water, the studied individuals use water from drilled wells containing elevated concentrations of natural uranium and its daughter nuclides ((234,235,238)U, 222Rn, (226,228)Ra, 210Po, 210Pb). Enhanced 210Pb and 235U activities were observed in several people. A positive correlation is observed between the U concentration in urine (microg d(-1)) and the number of counts (cpm) in the gamma ray energy peaks originating from the decay of 235U and 234Th respectively. Calibration of the detector set-up and the determination of background sources are in progress.     

Betel quid chewing elevates human exposure to arsenic, cadmium and lead

Several studies have reported increased skin lesions in betel quid (a mixture of Piper betel leaves, areca nut, tobacco/flavoured tobacco, lime) chewers compared to non-chewers, exposed to arsenic (As) contaminated drinking water in Bangladesh and India. The current study has determined As, cadmium (Cd) and lead (Pb) levels of betel quids and its components using inductively coupled plasma mass spectrometry (ICP-MS). The highest concentrations of As were found in slaked lime (4.56 mg kg(-1)) followed by Piper betel leaves (0.406 mg kg(-1)) and flavoured tobacco (zarda) (0.285 mg kg(-1)), with a mean concentrations of As in betel quids of 0.035 mg kg(-1) (SD 0.02 mg kg(-1)). Mean concentrations of Cd and Pb in ordinary quids were 0.028 (SD 0.07 mg kg(-1)) and 0.423 (SD 1.4 mg kg(-1)), respectively. We estimated that a daily intake of 6 betel quids could contribute 1.2, 1.9 and 8.5% of the provisional maximum tolerable daily intake (PMDTI) for As, Cd and Pb, respectively. Since betel quid chewing is most prevalent among women, our finding raises concern that women chewers - especially pregnant chewers - may be harming their health and that of their unborn babies through increased exposure to a mixture of toxic elements (As, Cd and Pb).     

Assessment of occupational exposure to magnetic fields in high-voltage substations (154/34.5 kV)

This work proposes the survey of magnetic field measurements taken in 154/31.5 kV substations in the city of Antalya for occupational exposure assessment. For measurements, three substations have been taken into account, where four occupants are present for each 8 h of shift, which means 12 occupants are present during the day. Operator desks at three different substations read a lowest magnetic field of 0.3 microT at minimum loaded season and reached up to 1 microT at maximum loaded season, which means operators exposed to a magnetic field of > or = 0.3 microT 8 h d(-1). In switch gear regions, maximum magnetic field reading is 23 microT at minimum loaded season and reached up to 70 microT. Outdoor magnetic field measurement at circuit breaker region goes up to 62 microT with respect to operator heights.     

Use of external magnetic fields to reduce reaction times in an immunoassay using micrometer-sized paramagnetic particles as labels (magnetoimmunoassay)

The paper presents a rapid immunoassay system capable of quantifying analyte in complex biological and environmental media. Antibody-coated micrometer-sized paramagnetic particles are used as labels in an assay in which they bind quantitatively with an analyte and capture antibody molecules immobilized on a polyester disk to form a sandwich assay. The assay is performed in a simple reaction vessel, and reactions between labels, analyte, and antibodies are accelerated by positioning magnets alternately above and below the vessel. The bound paramagnetic particles are quantified using a simple flat, spiral, coil located just below the polyester disk. The electronic circuitry associated with the coil uses components that are inexpensive and readily available. The coil has been designed to respond only to particles bound on the disk and not to unbound particles still in the test solution. Unbound particles are pulled away from the disk by the magnet before readings are taken. The use of the reaction vessel with the cardiac markers CRP and CKMB is described. No sample preparation or washing step is used in the assays, and results can be obtained in less than 3 min after introducing the sample into the vessel with sensitivities in the normal clinical range.     

Dose conversion coefficients for neutron exposure to the lens of the human eye

Dose conversion coefficients for the lens of the human eye have been calculated for neutron exposure at energies from 1 × 10(-9) to 20 MeV and several standard orientations: anterior-to-posterior, rotational and right lateral. MCNPX version 2.6.0, a Monte Carlo-based particle transport package, was used to determine the energy deposited in the lens of the eye. The human eyeball model was updated by partitioning the lens into sensitive and insensitive volumes as the anterior portion (sensitive volume) of the lens being more radiosensitive and prone to cataract formation. The updated eye model was used with the adult UF-ORNL mathematical phantom in the MCNPX transport calculations.     

An investigation of heat exchange in the human organism on exposure to internal heat

The thermal field of a person exposed to artificial heat is investigated. A variation in the blood flow as the temperature inside the body increases is calculated by solving the inverse problem of heat conductivity.Translated from Inzhenerno-Fizicheskii Zhurnal, Vol. 52, No. 4, pp. 654–659, April, 1987.     

Yttrium and lanthanides in human lung fluids, probing the exposure to atmospheric fallout

Inhalation of airborne particles can produce crystallization of phosphatic microcrysts in intraaveolar areas of lungs, sometimes degenerating into pulmonary fibrosis. Results of this study indicate that these pathologies are induced by interactions between lung fluids and inhaled atmospheric dust in people exposed to volcanic dust ejected from Mount Etna in 2001. Here, the lung solid-liquid interaction is evaluated by the distribution of yttrium and lanthanides (YLn) in fluid bronchoalveolar lavages on selected individuals according the classical geochemical approaches. We found that shale-normalised patterns of yttrium and lanthanides have a 'V shaped' feature corresponding to the depletion of elements from Nd to Tb when compared to the variable enrichments of heavy lanthanides, Y, La and Ce. These features and concurrent thermodynamic simulations suggest that phosphate precipitation can occur in lungs due to interactions between volcanic particles and fluids. We propose that patterns of yttrium and lanthanides can represent a viable explanation of some pathology observed in patients after prolonged exposure to atmospheric fallout and are suitable to become a diagnostic parameter of chemical environmental stresses.     

Verification of exposure to organophosphates: Generic mass spectrometric method for detection of human butyrylcholinesterase adducts

We present a generic mass spectrometric method to verify exposure to organophosphates, based on the chemical conversion of the phosphylated peptides obtained after pepsin digestion of human butyrylcholinesterase (HuBuChE) to a common precursor peptide. After exposure of plasma to various organophosphates (nerve agents, pesticides), HuBuChE was isolated from plasma by procainamide affinity-based solid-phase extraction. Upon subsequent pepsin digestion, the respective phosphylated nonapeptides could be identified in the digests. After treatment of the pepsin digests with Ba(OH)2 in the presence of a nucleophilic tag (a thiol or amine), the phosphylated nonapeptides were transformed into a common tagged nonapeptide that could be analyzed sensitively by means of LC tandem MS. So far, best results were obtained with 2-(3-aminopropylamino)ethanol as nucleophilic tag. By applying the presented method, HuBuChE inhibition can now be monitored accurately by mass spectrometry, without advance knowledge of the structure of the inhibitor.