Electrochemical and in vitro neuronal recording characteristics of multi-electrode arrays surface-modified with electro-co-deposited gold-platinum nanoparticles.

In order to complement the high impedance electrical property of gold nanoparticles (Au NPs) we have performed electro-co-deposition of gold-platinum nanoparticles (Au-Pt NPs) onto the Au multi-electrode array (MEA) and modified the Au-Pt NPs surface with cell adhesive poly-D-lysine via thiol chemistry based covalent binding. The Au-Pt NPs were analyzed to have bimetallic nature not the mixture of Au NPs and Pt NPs by X-ray diffraction analysis and to have impedance value (4.0 × 10(4) Ω (at 1 kHz)) comparable to that of Pt NPs. The performance of Au-Pt NP-modified MEAs was also checked in relation to neuronal signal recording. The noise level in Au-Pt NP-modified MEAs was lower than in that of Au NP-modified MEA.

Assessment of Image Quality of Coronary CT Angiography Using Deep Learning-Based CT Reconstruction: Phantom and Patient Studies.

BACKGROUND: In coronary computed tomography angiography (CCTA), the main issue of image quality is noise in obese patients, blooming artifacts due to calcium and stents, high-risk coronary plaques, and radiation exposure to patients. OBJECTIVE: To compare the CCTA image quality of deep learning-based reconstruction (DLR) with that of filtered back projection (FBP) and iterative reconstruction (IR). METHODS: This was a phantom study of 90 patients who underwent CCTA. CCTA images were acquired using FBP, IR, and DLR. In the phantom study, the aortic root and the left main coronary artery in the chest phantom were simulated using a needleless syringe. The patients were classified into three groups according to their body mass index. Noise, the signal-to-noise ratio (SNR), and the contrast-to-noise ratio (CNR) were measured for image quantification. A subjective analysis was also performed for FBP, IR, and DLR. RESULTS: According to the phantom study, DLR reduced noise by 59.8% compared to FBP and increased SNR and CNR by 121.4% and 123.6%, respectively. In a patient study, DLR reduced noise compared to FBP and IR. Furthermore, DLR increased the SNR and CNR more than FBP and IR. In terms of subjective scores, DLR was higher than FBP and IR. CONCLUSION: In both phantom and patient studies, DLR effectively reduced image noise and improved SNR and CNR. Therefore, the DLR may be useful for CCTA examinations.

Photopatterning of cell-adhesive-modified poly(ethyleneimine) for guided neuronal growth.

We describe photopatterning technique that employs the photodegradation of cell-adhesive-modified poly(ethyleneimine) (m-PEI) to fabricate precise micropatterns on the indium tin oxide (ITO) substrate for guided neuronal growth. The photodegradation of m-PEI coated on hydroxyl group-terminated ITO substrate created micropatterns over a large area through deep UV irradiation. The photopatterned m-PEI layer can effectively guide neurite outgrowth and control neurite extensions from individual neurons.

Selective binding and detection of magnetic labels using PHR sensor via photoresist micro-wells.

We have developed a novel platform for selective binding of magnetic labels on planar Hall resistance sensor (PHR) for biosensing applications. The photoresist (PR) micro wells were prepared on the PHR sensor junctions to trap the magnetic bead at specified locations on the sensor surface and thin layer of Au was sputtered in the PR wells immobilize bimolecular. The Au surface is functionalized with single-stranded oligonucleotide and further biotin was used to immobilize streptavidin coated magnetic labels (Dynabeads Myone 1.0 microm, Invitrogen Co.). After removal of the PR wells on the sensor surface the non specific binding magnetic labels were successfully removed and only the chemically bounded magnetic labels were remained on the Au surface for detection of biomolecules using PHR sensor. We controlled the number of magnetic labels on the PHR sensor surface by using different sizes of the PR well on the junctions. The specifically bounded magnetic labels were successfully detected by characterizing the individual PHR sensor junctions. This technique enables the complete control over the magnetic labels for selective binding of biomolecules on the sensor surface for increasing the sensitivity of the PHR sensor as well as removal of the non specific bindings on the sensor surface.

Excited-state intramolecular proton transfer on 2-(2'-hydroxy-4'-R-phenyl)benzothiazole nanoparticles and fluorescence wavelength depending on substituent and temperature.

The fluorescence emission properties of 2-(2'-hydroxy-4'-R-phenyl)benzothiazole (HBT-R) nanoparticles with different substituents (R = -COOH, -H, -CH(3), -OH, and -OCH(3)) were investigated using spectroscopic and theoretical methods. HBT-Rs displayed dual enol and keto (excited-state intramolecular proton transfer (ESIPT)) emissions in nonpolar solvents. The spectral change of their ESIPT emissions was affected differently by the electron donating (or withdrawing) power of the substituents; a bathochromic shift for the electron donating group and a hypsochromic shift in electron withdrawing group. In addition, the changes in energy levels calculated by the ab initio method were consistent with the spectral shifts of HBT-R in solution. We prepared aggregated HBT-R nanoparticles using a simple reprecipitation process in tetrahydrofuran-water solvents. The ESIPT emission of aggregated HBT-R nanoparticles was strongly enhanced (over 45 times) compared to those of monomer HBT-Rs in toluene, as markedly shifted ESIPT emissions are observed at longer wavelength without any quenching by self-absorption. Aggregated HBT-R nanoparticles showed longer lifetimes than those of monomer molecules. The temperature effect on the aqueous dispersion of the aggregated HBT-R nanoparticles was also explored. It shows a fluorescent ratiometric change in a range of temperature from 7 to 65 degrees C. A mechanism of a temperature-dependent equilibrium between the nanoparticles and the solvated enols is proposed for the emission color change.

Anti-oxidant effect of ascorbic and dehydroascorbic acids in hippocampal slice culture.

Ascorbic acid (AA) and dehydroascorbic acid (DHA) have been shown to have protective effects as anti-oxidants in experimental neurological disorder models such as stroke, ischemia, and epileptic seizures. The present study was conducted to examine the protective effects of AA and DHA on kainic acid (KA) neurotoxicity using organotypic hippocampal slice cultures. After 12h KA treatment, significant delayed neuronal death was detected in the CA3, but not the CA1, region. Pretreatment with intermediate doses of AA and DHA significantly prevented cell death and inhibited reactive oxygen species (ROS) level, and mitochondrial dysfunction in the CA3 region. In contrast, pretreatment with low or high doses of AA or DHA was not effective. These data suggest that pretreatment with both AA and DHA has dose-dependent neuroprotective effects on KA-induced neuronal injury through inhibiting ROS generation and mitochondrial dysfunction.

Centrifugal enhancement of Kaposi's sarcoma-associated virus infection of human endothelial cells in vitro.

In order to improve the efficiency of infection of primary human endothelial cells in vitro of Kaposi's sarcoma-associated herpesvirus (KSHV), the effect of low speed centrifugation was investigated. The recombinant KSHV, BAC36, was used to examine the centrifugal enhancement of KSHV. Infectivity was estimated by green fluorescent protein (GFP) expression and real-time RT-PCR. The enhancement of infectivity was dependent upon the time and force of centrifugation in human umbilical vein endothelial cells (HUVECs). Centrifugation enhanced the infectivity of KSHV by up to 70 fold compared to non-centrifugal control infection for the same period of time; viral mRNA expression was also enhanced by centrifugation. HUVECs that were centrifuged before infection with KSHV displayed no enhancement in infectivity; therefore, enhancement is believed to occur during centrifugation. In addition, the mechanisms of infection including the initial viral attachment to cells, lipid rafts, and clathrin-mediated and caveolae endocytosis appear to be similar in KSHV infection with and without centrifugal enhancement. These results show that low speed centrifugation could be a useful tool for improving the efficiency of KSHV infection in vitro.

In vitro extracellular recording and stimulation performance of nanoporous gold-modified multi-electrode arrays.

OBJECTIVE: Nanoporous gold (Au) structures can reduce the impedance and enhance the charge injection capability of multi-electrode arrays (MEAs) used for interfacing neuronal networks. Even though there are various nanoporous Au preparation techniques, fabrication of MEA based on low-cost electro-codeposition of Ag:Au has not been performed. In this work, we have modified a Au MEA via the electro-codeposition of Ag:Au alloy, followed by the chemical etching of Ag, and report on the in vitro extracellular recording and stimulation performance of the nanoporous Au-modified MEA. APPROACH: Ag:Au alloy was electro-codeposited on a bilayer lift-off resist sputter-deposition passivated Au MEA followed by chemical etching of Ag to form a porous Au structure. MAIN RESULTS: The porous Au structure was analyzed by scanning electron microscopy and tunneling electron microscopy and found to have an interconnected nanoporous Au structure. The impedance value of the nanoporous Au-modified MEA is 15.4 ± 0.55 kΩ at 1 kHz, accompanied by the base noise V rms of 2.4 ± 0.3 µV. The charge injection limit of the nanoporous Au-modified electrode estimated from voltage transient measurement is approximately 1 mC cm(-2), which is comparable to roughened platinum and carbon nanotube electrodes. The charge injection capability of the nanoporous Au-modified MEA was confirmed by observing stimulus-induced spikes at above 0.2 V. The nanoporous Au-modified MEA showed mechanical durability upon ultrasonic treatment for up to an hour. SIGNIFICANCE: Electro-codeposition of Ag:Au alloy combined with chemical etching Ag is a low-cost process for fabricating nanoporous Au-modified MEA suitable for establishing the stimulus-response relationship of cultured neuronal networks.

Fine neurite patterns from photocrosslinking of cell-repellent benzophenone copolymer.

We have synthesized photocrosslinkable benzophenone copolymer, Poly(St-co-MBz), and fabricated cell-repellent patterns of Poly(St-co-MBz) on covalently bound poly-D-lysine (PDL) layer via the photocrosslinking. We have successfully obtained fine grid line pattern with line width of 3 µm and fine neurite, presumably axon, patterns with excellent pattern fidelity. We found that benzophenone unit can be crosslinked under the exposure of UV (with the intensity of ∼77 mW/cm² at 280 nm and ∼60 mW/cm² at 365 nm) without photo-oxidative damage to PDL, poly-L-lysine, and polyethyleneimine.

Enhancement of neuronal cell adhesion by covalent binding of poly-D-lysine.

We have prepared the poly-d-lysine (PDL) bound surfaces for neuron cell culture by covalent binding between the poly-d-Lysine and substrates and investigated neuronal cell adhesion properties and cell growth morphology. The number of neuronal cell and the number of neurite per neuronal cell on PDL bound surfaces was much more than those on PDL coated surfaces and also the neuronal cells on PDL bounded surfaces survived a longer time. On the pattern of covalently bound PDL, neuronal cells and their neurites are confined within the grid line leading to patterned neuronal networks with the long-term survival.

Detection of two different influenza A viruses using a nitrocellulose membrane and a magnetic biosensor.

Here we describe a new analytical method for the detection of two influenza A viruses by nitrocellulose membrane and magnetic sensors that employ a special frequency mixing technique. The combination of the nitrocellulose membrane and magnetic bead detection permits a rapid assay procedure and excludes two steps (the development of color and the stop reaction) required for usual immunochemical detection methods such as ELISA. Quantitative virus detection was performed using magnetic beads conjugated with secondary antibody. The results were compared with conventional assay methods and with a dot-blot assay with fluorescence compound (FITC). Under optimum conditions, our new assay procedure is capable of detecting picograms of virus per well. This new method combining the nitrocellulose membrane and magnetic bead detection reduces analytical time and allows stable and repeatable analyses of samples in point-of-care applications.

Review of semantically interoperable electronic health records for ubiquitous healthcare.

In order to provide more effective and personalized healthcare services to patients and healthcare professionals, intelligent active knowledge management and reasoning systems with semantic interoperability are needed. Technological developments have changed ubiquitous healthcare making it more semantically interoperable and individual patient-based; however, there are also limitations to these methodologies. Based upon an extensive review of international literature, this paper describes two technological approaches to semantically interoperable electronic health records for ubiquitous healthcare data management: the ontology-based model and the information, or openEHR archetype model, and the link to standard terminologies such as SNOMED-CT.

Neuroprotective effects of mexiletine on motor evoked potentials in demyelinated rat spinal cords.

This study was conducted to whether the administration of mexiletine, a Na+ channel blocker, impacts the recovery from demyelination. Under anesthesia, 0.1% ethidium bromide was injected into the dorsal funiculus (T3), followed by a mexiletine or saline treatment. Motor evoked potential (MEP) recordings and luxol fast blue stainings were performed at one, seven, 14, and 21 days post-operatively. Conduction was delayed during demyelination, but the mexiletine-injected group demonstrated shortened latencies and reductions in the demyelination area when compared to the control. These results suggest that systemic mexiletine plays a positive role in protecting neural tissues from demyelination.

Neuroprotective effects of FK506 against excitotoxicity in organotypic hippocampal slice culture.

FK506 has been originally classified as an immunosuppressant and is known to exhibit neurotrophic actions in vitro and protective effects on some neurological conditions. We investigated the neuroprotective effects of FK506 on kainic acid (KA)-induced neuronal death in organotypic hippocampal slice cultures (OHSCs). After an 18 h KA (5 microM) treatment, significantly neuronal death was detected in the CA3 region using propidium iodide staining. However, neuronal death was significantly prevented at 24 and 48 h after treatment with 0.1 microM FK506. Using cresyl violet staining, we also observed that an increased number of CA3 neurons survived in the 0.1 microM FK506 group compared to the KA only group. Based on the results of the Western blot analysis, the expressions of 5-lipoxygenase and caspase-3 were reduced 24h after 0.1 microM FK506 treatment. The levels of superoxide dismutase (SOD) and phospho-Akt expression were increased by treatment with 0.1 microM FK506. These results suggest that FK506 may have a positive role in protecting neurons against cell death in the KA injury model of OHSCs.

Modification of cortical excitability in neuropathic rats: a voltage-sensitive dye study.

Recent advances in optical imaging techniques have made it possible to monitor neural activity and provided powerful tools to reveal the spatiotemporal patterns of neural activity. We used optical imaging to determine whether nerve injury affects excitability of the sensory cortex. Male Sprague-Dawley rats were subjected to neuropathic surgery consisting of a tight ligation and transection of the left tibial and sural nerves while under pentobarbital anesthesia. The rats were reanesthetized with urethane two weeks post-operatively, and the exposed cortex surfaces were stained with a voltage-sensitive dye (di-2-ANEPEQ). After electrical stimulation of the receptive field, optical signals from the cerebral cortex were recorded using an optical imaging system. Increased optical intensity and an enlarged area of activation were observed in the cerebral cortex of neuropathic rats during electrical stimulation compared to normal or sham-operated rats. Higher electric stimulation resulted in more intensity and a larger area of activation in neuropathic rats. These results suggest that cortical excitability, resulting from peripheral stimulation, may be affected by nerve injury, which indicates a degree of neural plasticity.