Polymer nanochannels fabricated by thermomechanical deformation for single-molecule analysis

A simple method for fabricating nanoscale channels based on thermomechanical deformation of rigid polymer substrates is demonstrated. Polycarbonate preforms containing microchannels with cross-sectional dimensions on the order of tens of micrometers are controllably deformed to produce submicrometer dimensions. The reduced channel dimensions are achieved by heating the preform while applying a uniaxial tensile force to reduce channel cross sections through the Poisson effect. Nanochannels with circular or elliptical cross sections are defined by varying the channel position and preform geometry prior to deformation. Arrays of parallel nanochannels with critical dimensions down to 400 nm are described. Using the fabrication method, a nanochannel network is fabricated for the detection of single protein molecules via confocal fluorescence microscopy. The chip includes a detection channel with cross-sectional dimensions approaching the confocal volume dimensions of the detection optics and a larger adjacent reference channel used to optimize focusing. Detection of fluorescently labeled bovine serum albumin at 15 and 150 nM concentrations is presented, demonstrating the ability to perform single-molecule fluorescence measurements in polycarbonate chips using visible wavelengths for excitation and detection.     

Important role of blood rheology in atherosclerosis of patients with hemodialysis

Concerning a role of blood rheology for atherosclerosis in patients with hemodialysis (HD), little data are available. It may be due to the fact that the method for evaluating rheologic properties of circulating blood has been limited. We examined blood rheology in 118 HD patients by using microchannel array flow analyzer that makes it possible to directly observe the flow of blood cell elements through the microchannel. Transit time (T(B)) of heparinized whole blood through slit pores (7 x 30 microm) was used as an index of rheology and related with various inflammatory biomarkers such as high-sensitive CRP (hsCRP), monocyte chemotactic protein-1, osteopontin, or fibrinogen (Fg). Moreover, as a surrogate marker of atherosclerosis, carotid intima-media thickness (IMT) and aortic stiffness evaluated by brachial-ankle pulse-wave velocity (baPWV) were studied. In HD patients, T(B) had strong positive correlations with hsCRP (r = 0.427; p < 0.00001), Fg (r = 0.452; p < 0.00001), and osteopontin (r = 0.227; p < 0.0134). Further, T(B) was significantly well correlated with IMT (r = 0.400; p < 0.0001) and PWV (r = 0.470; p < 0.0001). Multivariate regression analysis showed that baPWV, IMT, Fg, hematocrit, white blood cell count, and CRP were chosen as significant explanatory factors for T(B.) These results suggest that blood rheology may play an important role for atherosclerosis in patients with HD.     

Effect of gravity on InGaSb crystal growth

The effect of gravity on dissolution of GaSb in InSb melt and growth of InGaSb was experimentally investigated. Experiments were carried out in a GaSb(seed)/InSb/GaSb(feed) sandwich system under an imposed temperature gradient. In the experiments, the GaSb feed crystal dissolved into the InSb melt to supply the required GaSb component for the growth of In_0.1Ga_0.9Sb crystal. Two parameters were considered: (1) the inclination angle (θ) of the sample for gravity as 0° and 53°, and (2) the sample diameter (D) as 9 mm and 5mm. When θ was 0°, the interface was almost flat, indicating that convection was axisymmetric and stable. Whereas the interface was distorted towards gravitational direction when θ was 53°, indicating that solutal convection was dominant. The decrease of growth temperature and sample diameter reduced the distortion of interface and the dissolution amount of GaSb feed. The homogeneous crystals were grown at the initial growth stage by supplying the GaSb component during growth.     

Design for an aberration-corrected concave grating for a mid-infrared long-slit spectrometer

A new design for an aberration-corrected concave grating for the spectral region near 10 mum is presented. It was designed for use in the ground-based astronomical medium-resolution (lambda/Dlambda ~ 100) Mid-Infrared Camera and Spectrometer (MICS). It provides a flat focal plane for a wide spectral range (7.5-13.5 mum) with small aberrations, permitting efficient long-slit observations in the mid-infrared region. It permits a simple design of the spectrometer without collimator and camera mirrors, which is quite advantageous for cryogenic instruments. The grating has variable spacing grooves to reduce aberrations. In addition, the grating surface figure is designed to be toroidal and in the direction perpendicular to the grooves, aspherical, to suppress the aberrations further over a wide spectral range. The angle of the grooves is also varied to yield better efficiency near the blaze angle. The grating was fabricated by high-quality ultraprecision machining, which made these features possible. Test observations confirmed that the designed spectral resolution was achieved.     

Comparison of ciliary sulcus fixation techniques for posterior chamber intraocular lenses

OBJECTIVE: To anatomically and histologically evaluate suturing techniques for sulcus fixation of posterior chamber intraocular lenses. SETTING: Kitano Hospital, Osaka, Japan. METHODS: The reproducibility of three suturing techniques (perpendicular to the eye wall; parallel to the iris; midway between perpendicular to the eye wall and parallel to the iris) were evaluated in a postmortem eye. Histologic sections of another eye and ultrasound biomicroscopy (UBM) images of 21 normal eyes were evaluated to determine the safety zone for the needle to avoid damaging adjacent structures. RESULTS: The ciliary sulcus was completely penetrated in the three techniques in 100, 40, and 70% of cases, respectively. The histologic sections and the UBM images showed that when sutures were placed perpendicular to the eye wall, there was the possibility of postoperative angle closure and suturing parallel to the iris might damage adjacent structures because of a narrow safety zone. CONCLUSIONS: The needle should penetrate obliquely, as in the technique in which the suturing is midway between perpendicular to the eye wall and parallel to the iris. This technique provides better reproducibility and causes less damage to adjacent tissue.     

Basic fibroblast growth factor is neither necessary nor sufficient for the development of retinal neovascularization

Basic fibroblast growth factor (FGF2) is constitutively expressed in the retina and its expression is increased by a number of insults, but its role in the retina is still uncertain. This study was designed to test the hypothesis that altered expression of FGF2 in the retina affects the development of retinal neovascularization. Mice with targeted disruption of the Fgf2 gene had no detectable expression of FGF2 in the retina by Western blot, but retinal vessels were not different in appearance or total area from wild-type mice. When FGF2-deficient mice were compared with wild-type mice in a murine model of oxygen-induced ischemic retinopathy, they developed the same amount of retinal neovascularization. Transgenic mice with a rhodopsin promoter/Fgf2 gene fusion expressed high levels of FGF2 in retinal photoreceptors but developed no retinal neovascularization or other abnormalities of retinal vessels; in the ischemic retinopathy model, they showed no significant difference in the amount of retinal neovascularization compared with wild-type mice. These data indicate that FGF2 expression is not necessary nor sufficient for the development of retinal neovascularization. This suggests that agents that specifically antagonize FGF2 are not likely to be useful adjuncts in the treatment of retinal neovascularization and therapies designed to increase FGF2 expression are not likely to be complicated by retinal neovascularization.     

Suppression of intracellular resistance factors by adriamycin augments heat-induced apoptosis via interleukin-1beta-converting enzyme activation in pancreatic carcinoma cells

The liver is an important target organ for gene therapy but its mitotic quiescence makes it resistant to integrative gene transfer. Retrovirus-based vectors integrate into liver cells in vivo but require the liver to be primed before transduction; experimentally a 70% hepatectomy is commonly used to stimulate regeneration, rendering the liver susceptible to transduction during the resulting wave of cell proliferation. Our aim was to develop a clinically acceptable method of inducing hepatocyte replication before in vivo retroviral gene transfer which is both simple and effective. We have used the physiological hormone tri-iodothyronine (T3) to stimulate hepatocyte replication. A single dose of T3 (400 micrograms/100 g bw) was given subcutaneously to euthyroid rats. This produced a labelling index of 31.7% in the hepatocyte population without histological or biochemical evidence of preceding liver damage. Following T3 administration the rat livers were transfected in vivo with an amphotropic retrovirus, TELCeB/AF-7 which encodes the beta-galactosidase reporter gene together with a nuclear localisation signal. Transgene expression was noted only within the liver where 1.3% of hepatocytes expressed the beta-galactosidase enzyme. This compared to 5.2% of hepatocytes transduced following a 70% hepatectomy, and 0.02% in animals receiving neither T3 nor partial hepatic resection before transduction. T3 administration is a simple way to prime the liver before in vivo retroviral vector-based gene transfer.