Microfabricated silicon biosensors for microphysiometry

Microphysiometers are biosensor devices that measure the metabolic rate of living cells by detecting the rate of extracellular acidification caused by a small number of cells. The cells are entrapped in a microvolume chamber, whose bottom surface is a silicon sensor chip. In a further miniaturization step, we have recently fabricated multichannel flow-through chips that will allow greater throughput and multiplicity. Microphysiometer technology can be applied to the detection of microorganisms. We describe the sensitive detection of bacteria and yeast. Further applications of microphysiometry to the characterization of microorganisms can be anticipated

The isfet in analytical chemistry

The fast chemical response of the pH-ISFET makes the device an excellent detector in analytical chemistry. The time response of ISFETs, with Al2O3 at the pH-sensitive gate insulator, is determined in a flow injection analysis system. Application of an ISFET and a glass electrode are compared in rapid acid-base titrations and in a coulometric system for stable pH control. Finally, a method is described that combines both the stability of the glass electrode and the fast response of the ISFET

Collimator design for a clinical brain SPECT/MRI insert

This project's goal is to design a SPECT insert for a clinical MRI system for simultaneous brain SPECT/MR imaging. We assume the stationary SPECT insert will consist of two rings of ∼5x5-cm SiPM-based detectors insensitive to magnetic fields, with 0.8-mm intrinsic resolution. The maximum diameter is 44.5 cm, the minimum diameter is 33 cm to accommodate the patient and MRI receive/transmit coil, and the FOV has a 20 cm diameter. We have compared eight collimator designs: single-, 2x2-, 3x3- and 5+2½- pinhole, and single-, 2-, 3- and 1+2½-slit slit-slat, where ½-pinholes/slits are shared between two detectors. Analytical geometric efficiency was calculated for an activity distribution corresponding to a human brain and a target resolution of 10 mm FWHM at the centre of the FOV. Noise-free data were simulated with and without depth-of-interaction (DOI) information, and reconstructed for uniform, Defrise, Derenzo, and Zubal brain phantoms. For DOI it is assumed that the crystal's first and second half can be differentiated. Comparing the multi-pinhole and multi-slit slit-slat collimators, the former gives better reconstructed uniformity and trans-axial resolution, while the latter gives better axial resolution. Although the 2x2-pinhole and 2-slit designs give the highest sensitivities, they result in a sub-optimal utilization of the detector FOV. The best options are therefore the 5+2½-pinhole and the 1+2½-slit systems, with sensitivities of 4.9*10–4 and 4.0*10–4, respectively. The brain phantom reconstructions with multi-pinhole collimator are superior as compared to slit-slat, especially in terms of symmetry and realistic activity distribution. DOI information reduces artefacts and improves uniformity in geometric phantoms, although the difference is small for the brain phantom. These results favour a multi-pinhole configuration

Respiratory Motion Correction in Dynamic PET with a Single Attenuation Map

In addition to static tracer uptake values used routinely in clinical facilities, PET imaging can provide useful information on tracer kinetics via the use of dynamic acquisitions where a set of time frames are acquired starting from the injection/inhalation of the radiotracer. In lung studies, kinetic parameters, estimated from compartmental modelling, are however affected by respiratory motion. When only one attenuation image is available, most existing motion compensation strategies are not appropriate for the initial short time frames, especially as the activity distribution changes rapidly over the early part of the dynamic acquisition. This work presents a preliminary study to handle respiratory motion using a two-step process that uses gated dynamic data as input. We first use joint reconstruction of activity and motion on the entire gated PET data to estimate deformation fields. This allows the subsequent reconstruction of each time frame separately with motion compensation. We present results comparing on one hand the compartment model fit residuals with and without respiratory motion compensation and on the other hand the diaphragm position in non-attenuation corrected images and from this method

Detection of a Functional Hybrid Receptor γc/GM-CSFRβ in Human Hematopoietic CD34+ Cells

A functional hybrid receptor associating the common γ chain (γc) with the granulocyte/macrophage colony-stimulating factor receptor β (GM-CSFRβ) chain is found in mobilized human peripheral blood (MPB) CD34+ hematopoietic progenitors, SCF/Flt3-L primed cord blood (CB) precursors (CBPr CD34+/CD56−), and CD34+ myeloid cell lines, but not in normal natural killer (NK) cells, the cytolytic NK-L cell line or nonhematopoietic cells. We demonstrated, using CD34+ TF1β cells, which express an interleukin (IL)-15Rα/β/γc receptor, that within the hybrid receptor, the GM-CSFRβ chain inhibits the IL-15–triggered γc/JAK3-specific signaling controlling TF1β cell proliferation. However, the γc chain is part of a functional GM-CSFR, activating GM-CSF–dependent STAT5 nuclear translocation and the proliferation of TF1β cells. The hybrid receptor is functional in normal hematopoietic progenitors in which both subunits control STAT5 activation. Finally, the parental TF1 cell line, which lacks the IL-15Rβ chain, nevertheless expresses both a functional hybrid receptor that controls JAK3 phosphorylation and a novel IL-15α/γc/TRAF2 complex that triggers nuclear factor κB activation. The lineage-dependent distribution and function of these receptors suggest that they are involved in hematopoiesis because they modify transduction pathways that play a major role in the differentiation of hematopoietic progenitors

Graphene transistors are insensitive to pH changes in solution

We observe very small gate-voltage shifts in the transfer characteristic of as-prepared graphene field-effect transistors (GFETs) when the pH of the buffer is changed. This observation is in strong contrast to Si-based ion-sensitive FETs. The low gate-shift of a GFET can be further reduced if the graphene surface is covered with a hydrophobic fluorobenzene layer. If a thin Al-oxide layer is applied instead, the opposite happens. This suggests that clean graphene does not sense the chemical potential of protons. A GFET can therefore be used as a reference electrode in an aqueous electrolyte. Our finding sheds light on the large variety of pH-induced gate shifts that have been published for GFETs in the recent literature

Collimator Design for a Brain SPECT/MRI Insert

This project's goal is to design a SPECT insert for a clinical MRI system for simultaneous brain SPECT/MR imaging, with a high-sensitivity collimator and high-resolution detectors. We have compared eight collimator designs, four multi-pinhole and four multi-slit slit-slat configurations. The collimation was designed for a system with 2 rings of 25 5 × 5 cm detectors. We introduce the concept of 1/2-pinhole and 1/2-slit, which are transaxially shared between two adjacent detectors. Analytical geometric efficiency was calculated for an activity distribution corresponding to a human brain and a range of intrinsic detector resolutions Ri and target resolutions Rt at the centre of the FOV. Noise-free data were simulated with and without depth-of-interaction (DOI) information, 0.8 mm Ri and 10 mm Rt FWHM, and reconstructed for uniform, Defrise, Derenzo, and Zubal brain phantoms. Comparing the multi-pinhole and multi-slit slit-slat collimators, the former gives better reconstructed uniformity and transaxial resolution, while the latter gives better axial resolution. Although the 2 ×2-pinhole and 2-slit designs give the highest sensitivities, they result in a sub-optimal utilisation of the detector FOV. The best options are therefore the 5+ 2 1/2-pinhole and the 1 + 2 1/2-slit systems, with sensitivities of 1.8 ×10-4 and 3.2 ×10-4, respectively. Noiseless brain phantom reconstructions with the multi-pinhole collimator are slightly superior as compared to slit-slat, in terms of symmetry and accuracy of the activity distribution, but the same is not true when noise is included. DOI information reduces artefacts and improves uniformity in geometric phantoms. Further evaluation is needed with prototype collimators

Stepwise Optimization of the Procedure for Assessment of Circulating Progenitor Cells in Patients with Myocardial Infarction

The number and functional activity of circulating progenitor cells (CPCs) is altered in diabetic patients. Furthermore, reduced CPC count has been shown to independently predict cardiovascular events. Validation of CPCs as a biomarker for cardiovascular risk stratification requires rigorous methodology. Before a standard operation protocol (SOP) can be designed for such a trial, a variety of technical issues have to be addressed fundamentally, which include the appropriate type of red blood cell lysis buffer, FMO or isotype controls to identify rare cell populations from background noise, optimal antibody dilutions and conditions of sample storage. We herein propose improvements in critical steps of CPC isolation, antigenic characterization and determination of functional competence for final application in a prospective investigation of CPCs as a biomarker of outcome following acute myocardial infarction.In this validation study, we refined the standard operating procedure (SOP) for flow cytometry characterisation and functional analysis of CPCs from the first 18 patients of the Progenitor Cell Response after Myocardial Infarction Study (ProMIS). ProMIS aims to verify the prognostic value of CPCs in patients with either ST elevation or non-ST elevation myocardial infarction with or without diabetes mellitus, using cardiac magnetic resonance imaging (MRI) for assessment of ventricular function as a primary endpoint. Results indicate crucial steps for SOP implementation, namely timely cell isolation after sampling, use of appropriate lysis buffer to separate blood cell types and minimize the acquisition events during flow cytometry, adoption of proper fluorophore combination and antibody titration for multiple antigenic detection and introduction of counting beads for precise quantification of functional CPC activity in migration assay.With systematic specification of factors influencing the enumeration of CPC by flow cytometry, the abundance and migration capacity of CPCs can be correctly assessed. Adoption of validated SOP is essential for refined comparison of patients with different comorbidities in the analysis of risk stratification

Functional significance of the hemadsorption activity of influenza virus neuraminidase and its alteration in pandemic viruses

Human influenza viruses derive their genes from avian viruses. The neuraminidase (NA) of the avian viruses has, in addition to the catalytic site, a separate sialic acid binding site (hemadsorption site) that is not present in human viruses. The biological significance of the NA hemadsorption activity in avian influenza viruses remained elusive. A sequence database analysis revealed that the NAs of the majority of human H2N2 viruses isolated during the influenza pandemic of 1957 differ from their putative avian precursor by amino acid substitutions in the hemadsorption site. We found that the NA of a representative pandemic virus A/Singapore/1/57 (H2N2) lacks hemadsorption activity and that a single reversion to the avian-virus-like sequence (N367S) restores hemadsorption. Using this hemadsorption-positive NA, we generated three NA variants with substitutions S370L, N400S and W403R that have been found in the hemadsorption site of human H2N2 viruses. Each substitution abolished hemadsorption activity. Although, there was no correlation between hemadsorption activity of the NA variants and their enzymatic activity with respect to monovalent substrates, all four hemadsorption-negative NAs desialylated macromolecular substrates significantly slower than did the hemadsorption-positive counterpart. The NA of the 1918 pandemic virus A/Brevig Mission/1/18 (H1N1) also differed from avian N1 NAs by reduced hemadsorption activity and less efficient hydrolysis of macromolecular substrates. Our data indicate that the hemadsorption site serves to enhance the catalytic efficiency of NA and they suggest that, in addition to changes in the receptor-binding specificity of the hemagglutinin, alterations of the NA are needed for the emergence of pandemic influenza viruses