Analysis of Sus scrofa liver proteome and identification of proteins differentially expressed between genders, and conventional and genetically enhanced lines.

Porcine liver proteome iTRAQ analysis enabled the confident identification of 880 proteins with a rate of false positive identifications of less than 5%. Proteins involved in energy metabolism, catabolism, protein biosynthesis, electron transport, and other oxidoreductase reactions were highly enriched confirming the central role of liver as the major chemical and energy factory. Comparative analysis with human and mouse liver proteomes demonstrated that 80% of proteins were common to all three liver proteomes.

Glucose-6-phosphate dehydrogenase deficiency enhances enterovirus 71 infection.

Variations in the cellular microenvironment affect the host's susceptibility to pathogens. Using glucose-6-phosphate dehydrogenase (G6PD)-deficient fibroblasts as a model, this study demonstrated that the cellular redox status affects infectivity as well as the outcome of enterovirus 71 (EV71) infection. Compared with their normal counterparts, G6PD-deficient cells supported EV71 replication more efficiently and showed greater cytopathic effect and loss of viability.

Calcium-assisted glass-to-glass bonding for fabrication of glass microfluidic devices.

Glass is a desired material for many microfluidics applications. It is chemically resistant and has desirable characteristics for capillary electrophoresis. The process to make a glass chip, however, is lengthy and inconvenient, with the most difficult step often being the bonding of two planar glass substrates.

Single-synapse ablation and long-term imaging in live C. elegans.

Synapses are individually operated, computational units for neural communication. To manipulate physically individual synapses in a living organism, we have developed a laser ablation technique for removing single synapses in live neurons in C. elegans that operates without apparent damage to the axon.

Microarray for molecular typing of Salmonella enterica serovars.

We describe the development of a spotted array for the delineation of the most common 14 disease-causing Salmonella serovars in the United States. Our array consists of 414 70 mers targeting core genes of Salmonella enterica, subspecies I specific genes, fimbrial genes, pathogenicity islands, Gifsy elements and other variable genes. Using this array we were able to identify a unique gene presence/absence profile for each of the targeted serovar which was used as the serovar differentiating criteria.

Multipattern consensus regions in multiple aligned protein sequences and their segmentation.

Decomposing a biological sequence into its functional regions is an important prerequisite to understand the molecule. Using the multiple alignments of the sequences, we evaluate a segmentation based on the type of statistical variation pattern from each of the aligned sites. To describe such a more general pattern, we introduce multipattern consensus regions as segmented regions based on conserved as well as interdependent patterns.

Sizing subcellular organelles and nanoparticles confined within aqueous droplets.

This article describes two complementary techniques, single-particle tracking and correlation spectroscopy, for accurately sizing nanoparticles confined within picoliter volume aqueous droplets. Single-particle tracking works well with bright particles that can be continuously illuminated and imaged, and we demonstrated this approach for sizing single fluorescent beads. Fluorescence correlation spectroscopy detects small intensity bursts from particles or molecules diffusing through the confocal probe volume, which works well with dim and rapidly diffusing particles or molecules; we demonstrated FCS for sizing synaptic vesicles confined in aqueous droplets.

Alzheimer's disease protein Abeta1-42 does not disrupt isolated synaptic vesicles.

Synaptic vesicles are central to neurotransmission and cognition. Studies of the Alzheimer's disease (AD) associated peptide, amyloid beta (Abeta), suggest that it has the potential to non-specifically solubilize or permeabilize membranes and that it has detergent and pore-forming properties. Damage to the membrane or integrity of synaptic vesicles could compromise its function.

Elevated levels of myeloperoxidase, white blood cell count and 3-chlorotyrosine in Taiwanese patients with acute myocardial infarction.

Objectives: Inflammation, a major risk factor for acute myocardial infarction (AMI), is associated with leukocytic activation, secretion of myeloperoxidase (MPO) and generation of the oxidative damage marker, 3-chlorotyrosine (3-Cl-Tyr). To study their association with AMI and their value in diagnosis of AMI, white blood cell (WBC) count, plasma MPO, plasma 3-Cl-Tyr, and conventional risk factors such as cardiac troponin I and CK-MB were examined in AMI patients during the onset of chest pain.

Methods: After obtaining informed consent, blood samples were collected from 77 AMI patients during the onset of chest pain and from 53 normal controls.

Glucose-6-phosphate dehydrogenase deficiency enhances human coronavirus 229E infection.

The host cellular environment is a key determinant of pathogen infectivity. Viral gene expression and viral particle production of glucose-6-phosphate dehydrogenase (G6PD)-deficient and G6PD-knockdown cells were much higher than their counterparts when human coronavirus (HCoV) 229E was applied at 0.1 multiplicity of infection.

Liquid chromatography-mass spectrometry in metabolite profiling.

Recent advances in the use of liquid chromatography-mass spectrometry for the study of metabolomics are reviewed. Sample preparations of biofluids and practical aspects of ultra-high pressure liquid chromatography are discussed. Applicability of different kinds of mass spectrometers for metabolite profiling is described.

Oxidative damage markers and antioxidants in patients with acute myocardial infarction and their clinical significance.

Oxidative stress has been associated with degenerative diseases such as cardiovascular and neurodegenerative diseases. Acute myocardial infarction (AMI) is the major cause of death among cardiovascular diseases. Inflammation, a major risk factor of AMI, is associated with leukocytic activation, secretion of myeloperoxidase (MPO) and subsequent oxidant generation.

Clinical relevance of plasma homocysteine levels in Taiwanese patients with coronary artery disease.

Homocysteine (Hcy) has recently been considered as a risk factor for cardiovascular disease. Coronary artery disease (CAD) is the third most common cause of mortality in Taiwan. The objective of our present study is to delineate the relationship between the plasma Hcy level and CAD in Taiwanese using an improved enzymatic method.

Measuring the functional sequence complexity of proteins.

Background: Abel and Trevors have delineated three aspects of sequence complexity, Random Sequence Complexity (RSC), Ordered Sequence Complexity (OSC) and Functional Sequence Complexity (FSC) observed in biosequences such as proteins. In this paper, we provide a method to measure functional sequence complexity.

Methods And Results: We have extended Shannon uncertainty by incorporating the data variable with a functionality variable.

Spin-to-orbital angular momentum conversion in a strongly focused optical beam.

As a fundamental property of light, the angular momentum of photons has been of great interest. Here, we demonstrate that optical spin-to-orbital angular momentum conversion can occur in a homogeneous and isotropic medium. This Letter presents both theoretical and experimental studies of this conversion in a tightly focused beam and shows that the orbital rotation speeds of trapped particles are altered because of this conversion as predicted by theory.

Fourier transform capillary electrophoresis with laminar-flow gated pressure injection.

Fourier transform capillary electrophoresis (FTCE) was developed as a method to improve signal-to-noise ratio (S/N) and resolution in capillary electrophoresis (CE) separation. In FTCE, multiple simultaneous CE separations were performed in the same channel system and interrogated using a single-point detector. To illustrate experimentally the improvement offered by FTCE in S/N ratio and resolution, we carried out a modest number (five) of multiple injections and separations.

In-channel atom-transfer radical polymerization of thermoset polyester microfluidic devices for bioanalytical applications.

A new technique for polymer microchannel surface modification, called in-channel atom-transfer radical polymerization, has been developed and applied in the surface derivatization of thermoset polyester (TPE) microdevices with poly(ethylene glycol) (PEG). X-ray photoelectron spectroscopy, electroosmotic flow (EOF), and contact angle measurements indicate that PEG has been grafted on the TPE surface. Moreover, PEG-modified microchannels have much lower and more pH-stable EOF, more hydrophilic surfaces and reduced nonspecific protein adsorption.

Glucose-6-phosphate dehydrogenase--from oxidative stress to cellular functions and degenerative diseases.

Glucose-6-phosphate dehydrogenase (G6PD), the first and rate-limiting enzyme of the pentose phosphate pathway, is indispensable to maintenance of the cytosolic pool of NADPH and thus the cellular redox balance. The role of G6PD as an antioxidant enzyme has been recognized in erythrocytes for a long time, as its deficiency is associated with neonatal jaundice, drug- or infection-mediated hemolytic crisis, favism and, less commonly, chronic non-spherocytic hemolytic anemia. To a large extent, advances in the field were made on the pathophysiology of G6PD-deficient erythrocytes, and the molecular characterization of different G6PD variants.

Fabrication improvements for thermoset polyester (TPE) microfluidic devices.

Thermoset polyester (TPE) microfluidic devices were previously developed as an alternative to poly(dimethylsiloxane) (PDMS) devices, fabricated similarly by replica molding, yet offering stable surface properties and good chemical compatibility with some organics that are incompatible with PDMS. This paper describes a number of improvements in the fabrication of TPE chips. Specifically, we describe methods to form TPE devices with a thin bottom layer for use with high numerical aperture (NA) objectives for sensitive fluorescence detection and optical manipulation.

Continuous-flow single-molecule CE with high detection efficiency.

This paper describes the use of two-beam line-confocal detection geometry for measuring the total mobility of individual molecules undergoing continuous-flow CE separation. High-sensitivity single-molecule confocal detection is usually performed with a diffraction limited focal spot (approximately 500 nm in diameter), which necessitates the use of nanometer-sized channels to ensure all molecules flow through the detection volume. To allow for the use of larger channels that are a few micrometers in width, we employed cylindrical optics to define a rectangular illumination area that is diffraction-limited (approximately 500 nm) in width, but a few micrometers in length to match the width of the microchannel.

Increased oxidative damage and mitochondrial abnormalities in the peripheral blood of Huntington's disease patients.

Increased oxidative stress and mitochondrial abnormalities contribute to neuronal dysfunction in Huntington's disease (HD). We investigated whether these pathological changes in HD brains may also be present in peripheral tissues. Leukocyte 8-hydroxydeoxyguanosine (8-OHdG) and plasma malondialdehyde (MDA) were elevated, and activities of erythrocyte Cu/Zn-superoxide dismutase (Cu/Zn-SOD) and glutathione peroxidase (GPx) reduced in 16 HD patients when compared to 36 age- and gender-matched controls.

Thermoelectric manipulation of aqueous droplets in microfluidic devices.

This article describes a method for manipulating the temperature inside aqueous droplets, utilizing a thermoelectric cooler to control the temperature of select portions of a microfluidic chip. To illustrate the adaptability of this approach, we have generated an "ice valve" to stop fluid flow in a microchannel. By taking advantage of the vastly different freezing points for aqueous solutions and immiscible oils, we froze a stream of aqueous droplets that were formed on-chip.

Optical gradient flow focusing.

This paper describes a new method for carrying out flow cytometry, which employs optical gradient forces to guide and focus particles in the fluid flow. An elliptically shaped Gaussian beam was focused at the center of a microchannel to exert radiation pressure on suspended nanoparticles that are passing through the channel, such that these particles are guided to the center of the channel for efficient detection and sorting. To verify the efficiency of this optical-gradient-flow-focusing method, we present numerical simulations of the trajectories of the nanoparticles in both electroosmotic flow (EOF) and pressure-driven flow (PDF).