Characterisation of optically driven microstructures for manipulating single DNA molecules under a fluorescence microscope

Optical tweezers are powerful tools for manipulating single DNA molecules using fluorescence microscopy, particularly in nanotechnology‐based DNA analysis. We previously proposed a manipulation technique using microstructures driven by optical tweezers that allows the handling of single giant DNA molecules of millimetre length that cannot be manipulated by conventional techniques. To further develop this technique, the authors characterised the microstructures quantitatively from the view point of fabrication and efficiency of DNA manipulation under a fluorescence microscope. The success rate and precision of the fabrications were evaluated. The results indicate that the microstructures are obtained in an aqueous solution with a precision ∼50 nm at concentrations in the order of 10⁶ particles/ml. The visibility of these microstructures under a fluorescence microscope was also characterised, along with the elucidation of the fabrication parameters needed to fine tune visibility. Manipulating yeast chromosomal DNA molecules with the microstructures illustrated the relationship between the efficiency of manipulation and the geometrical shape of the microstructure. This report provides the guidelines for designing microstructures used in single DNA molecule analysis based on on‐site DNA manipulation, and is expected to broaden the applications of this technique in the future.Inspec keywords: DNA, molecular biophysics, fluorescence, optical microscopy, radiation pressure, biological techniquesOther keywords: optically driven microstructures, single DNA molecule analysis, fluorescence microscopy, optical tweezers, nanotechnology‐based DNA analysis, manipulation technique, aqueous solution, fine tune visibility, yeast chromosomal DNA molecules, geometrical shape, on‐site DNA manipulation     

High-pressure studies on Tc and crystal structure of iron chalcogenide superconductors

The superconducting transition temperature, T_c, in iron-based solids can be enhanced by applied pressure: T_c increases from 8 to 37 K for the 11-type FeSe when the pressure is raised from 0 to 4 GPa. High-pressure studies can elucidate the mechanism of superconductivity in such novel materials. In this paper, we present a high-pressure study of Fe(Se_1−xTe_x) and Fe(Se_1−xS_x). In the case of Fe(Se_1−xTe_x), the maximum T_c under high pressure did not exceed the T_c of FeSe, which can be attributed to the structural transition to the monoclinic phase. For Fe(Se_1−xS_x) (0 < x < 0.3), T_c exhibited a significant increase with pressure; however, the maximum T_c under high pressure did not exceed the T_c of FeSe. This may be due to the disorder induced by substituting S for Se, which is similar to the pressure effect on T_c for the 1111-type superconductor Ca(Fe_1−xCo_x)AsF. The T_c of Fe(Se_1−xS_x) showed a complex behavior below 1 GPa, first decreasing and then increasing with increasing pressure. From high-pressure x-ray diffraction measurements, the T_c (P) curve was correlated with the local structural parameter.     

Synthesis of New Superconducting Pb-Based 1222 Cuprates Containing Sulfur in the (Pb0.75S0.25)Sr2(Eu2.0−x Ce x )Cu2O z System

We have synthesized new Pb-based 1222 cuprates containing sulfur in the (Pb_0.75S_0.25)Sr₂(Eu_2.0?x Ce _x )Cu₂O _z system. X-ray diffraction study shows that almost the single 1222 phase samples can be obtained for a considerable wide area of 0.3≤x≤1.2 in the composition of (Pb_0.75S_0.25)Sr₂(Eu_2.0?x Ce _x )Cu₂O _z . Next, the sample with x=0.4 after annealing under high-pressure O₂ is discovered to show an onset of the resistivity drop at about 31 K and to show an onset of diamagnetic signal at about 29 K. These phenomena clearly indicate that the sample is a new superconductor.     

New superconducting lead cuprate with 1201 structure, (Pb0.5B0.5) (SrLa)CuOz

New Pb-based superconducting cuprate with the 1201 structure has been synthesized in the (Pb_1 − yB_y)(Sr_2 − xLa_x)CuO_z system. From X-ray powder diffraction study, the almost-single 1201 phase sample is found to be obtained at x = 1.0 and y = 0.5 for the nominal composition of (Pb_1 − yB_y)(Sr_2 − xLa_x)CuO_z. The crystal structure has a tetragonal symmetry with the lattice parameters of a = 0.3779 nm and c = 0.8654 nm. The sample shows an onset of superconductivity at about 32 K as measured by the temperature dependence of the DC magnetic susceptibility.     

The effects of propofol, isoflurane, and sevoflurane on oxygenation and shunt fraction during one-lung ventilation

Propofol's effect on hypoxic pulmonary vasoconstriction during one-lung ventilation (OLV) has not been determined. Twenty patients who had long-term OLV for esophageal surgery were allocated randomly to one of two study groups; one in which isoflurane administration preceded propofol, and another in which sevoflurane administration preceded propofol. Arterial and mixed venous blood samples and hemodynamics were measured as follows: before OLV, during OLV, OLV at 4 cm of positive end-expiratory pressure (PEEP), OLV after conversion from volatile anesthetics to propofol, OLV at 4 cm of PEEP, and after OLV. After the application of 4 cm of PEEP during propofol anesthesia, PaO2 increased significantly in both groups. The shunt fraction (Qs/Qt) increased significantly after the initiation of OLV in both groups and decreased significantly after the conversion from volatile anesthetics to propofol in both groups. Propofol can be used safely during OLV because PaO2 increased after the application of 4 cm of PEEP during propofol anesthesia, and Qs/Qt decreased significantly after the conversion from inhaled anesthetics to propofol anesthesia. IMPLICATIONS: During one-lung ventilation, the arterial partial pressure of oxygen values with propofol were greater than those with isoflurane and sevoflurane, and shunt fraction values with propofol were lower than those with both volatile anesthetics. Propofol improved oxygenation and shunt fraction during one-lung ventilation compared with volatile anesthetics.     

HPLC method for evaluation of the free radical-scavenging activity of foods by using 1,1-diphenyl-2-picrylhydrazyl

An HPLC method for evaluation of the free radical-scavenging activity of foods by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) is reported. The activity was evaluated by measuring the decrease of DPPH detected at 517 nm. By using this novel method, we determined the free radical-scavenging activity of several antioxidants: ascorbic acid, alpha-tocopherol, Trolox, and cysteine. The results gave good correlation between the radical-scavenging activity determined by HPLC and by conventional colorimetry. This methodology was applied to determine the free radical-scavenging activity of 8 beverages. The activity of coffee was the highest, followed by red wine, green tea, oolong tea, black tea, rosé wine, white wine, and orange juice. The results well agree with those of previous reports. This method is expected to be useful for a simple and rapid determination of free radical-scavenging activity in colored foods, because coloring substances in foods do not interfere with the measurement.     

Functional mapping of pain-related activation with echo-planar MRI: significance of the SII-insular region

Activation in numerous regions of the brain is likely to be involved in the complex neural network function of pain perception. To detect the cortical representation during nonpainful and painful stimuli, which were presented using electrical finger stimulation in six normal right-handed male volunteers, we performed echo-planar functional magnetic resonance imaging (fMRI). Using a 1.5-T MR system that scanned the supratentorial region of the brain, we obtained multislice BOLD-based functional MR images with single-shot gradient-echo echo-planar imaging (EPI). The data show that dispersed brain regions are activated during painful stimulation, and especially demonstrate the significance of the SII-insular region in pain perception.     

beta-carotene 15,15'-dioxygenase activity and cellular retinol-binding protein type II level are enhanced by dietary unsaturated triacylglycerols in rat intestines

The purpose of this study was to examine effects of dietary triacylglycerols on beta-carotene 15,15'-dioxygenase (EC 1.13.11.21) activity and cellular retinol-binding protein [CRBP (II)] in rats. Six groups of eight rats (7-wk old) were fed one of the following diets: standard (STD; 2.5% soybean oil), saturated (SFA; 15% hydrogenated soybean oil), monounsaturated (MUFA; 15% olive oil), polyunsaturated (PUFA; 15% soybean oil) or clofibrate (CLF; 2.5% soybean oil + 0.2% clofibrate) for 3 wk. The dioxygenase specific activities of the intestinal homogenates in the MUFA and PUFA groups fed the high fat diets were 2.4 times that of the STD group fed a low fat diet (P < 0.01), whereas the activities of the SFA and CLF groups were not significantly different from that of the STD group. The level of CRBP (II) in the intestine of the PUFA group was 1. 3-fold that of the STD group (P < 0.05), whereas there were no significant differences among the other groups. In a second experiment, the dioxygenase activity of rat intestine was followed over 3 wk of feeding the STD and PUFA diets. After the PUFA diet was consumed for 1 d, the activity was enhanced to 2.7 times the baseline level and remained thereafter at that high level, whereas the activity of the STD group remained at the low baseline level. Thus, dietary polyunsaturated triacylglycerols enhanced both beta-carotene 15,15'-dioxygenase activity and CRBP (II) level in rat intestine. These results suggest that the dioxygenase and CRBP (II) are regulated by the same mechanism involving long-chain fatty acids and their metabolites.     

Visualization of the information flow through human oculomotor cortical regions by transcranial magnetic stimulation

We investigated the topography of human cortical activation during an antisaccade task by focal transcranial magnetic stimulation (TMS). We used a figure-eight shaped coil, with the stimulus intensity set just above the threshold for activation of the hand motor areas but weak enough not to elicit blinks. TMS was delivered at various time intervals (80, 100, and 120 ms) after target presentation over various sites on the scalp while the subjects performed the antisaccade task. It was possible to elicit a mild but significant delay in saccade onset over 1) the frontal regions (a region 2-4 cm anterior and 2-4 cm lateral to hand motor area) and 2) posterior parietal regions (6-8 cm posterior and 0-4 cm lateral to hand motor area) regardless of which hemisphere was stimulated. The frontal regions were assumed to correspond to a cortical region including the frontal eye fields (FEFs), whereas the parietal regions were assumed to represent a wide region that includes the posterior parietal cortices (PPCs). The regions inducing the delay shifted from the posterior parietal regions at an earlier interval (80 ms) to the frontal regions at a later interval (100 ms), which suggested an information flow from posterior to anterior cortical regions during the presaccadic period. At 120 ms, the effect of TMS over the frontal regions still persisted but was greatly diminished. Erroneous prosaccades to the presented target were elicited over a wide cortical region including the frontal and posterior parietal regions, which again showed a forward shift with time. However, the distribution of effective regions exhibited a clear contralateral predominance in terms of saccade direction. Our technique provides a useful method not only for detecting the topography of cortical regions active during saccadic eye movement, but also for constructing a physiological map to visualize the temporal evolution of functional activities in the relevant cortical regions.