mRNA analysis of single living cells

Analysis of specific gene expression in single living cells may become an important technique for cell biology. So far, no method has been available to detect mRNA in living cells without killing or destroying them. We have developed here a novel method to examine gene expression of living cells using an atomic force microscope (AFM). AFM tip was inserted into living cells to extract mRNAs. The obtained mRNAs were analyzed with RT-PCR, nested PCR, and quantitative PCR. This method enabled us to examine time-dependent gene expression of single living cells without serious damage to the cells

Spin-splitting in an AlxGa1−xN/GaN nanowire for a quantum-ring interferometer

[[abstract]]The authors thank M. H. Gau, C. H. Hsieh, C. C Yang, R. Y. Su, and C. H. Chen for their assistance. The project was supported by National Science Council and Core Facilities Laboratory in Kaohsiung-Pingtung Area, Taiwan. The authors are also grateful to W. C. Mitchel and M. Mah for their help and AFOSR/AOARD, USAF for financial support under Grant No. FA4869-07-1-4022.[[abstract]]An Al0.18 Ga0.82 N/GaN heterostructure was used to fabricate a ballistic nanowire with a wire width of 200 nm by focused ion beam. We observed the beating Shubnikov-de Haas oscillations in the nanowire with a spin-splitting energy of (2.4±0.3) meV. Based on the results, we proposed a spin-Hall quantum-ring interferometer made of Alx Ga1-x N/GaN nanowires for spintronic applications. © 2008 American Institute of Physics.[[booktype]]紙

Atomic Force Microscopy of Humic Acids

Atomic force microscopic (AFM) images of humic acids show discrete, globular particles, where particles of the order of magnitude 100 to 300 nm dominate the image fields; the humic acids had been grown to a steady state at pH 5.0. The AFM data are consistent with our previously reported small-angle neutron scattering (SANS) study done under similar conditions. In further agreement, the cluster-cluster interactions shown in our previous SANS study may have their counterparts in closely interacting particles appearing as twin particles in the AFM images

Cost-Effectiveness Analysis of Omalizumab for the Treatment of Severe Asthma in Japan and the Value of Responder Prediction Methods Based on a Multinational Trial

AbstractObjectivesOmalizumab improves health outcomes for patients with severe asthma. The purpose of this study was to conduct a cost-utility analysis of omalizumab from a societal perspective by using the results from a randomized controlled trial in Japan, and explore the efficient use of omalizumab.MethodsWe developed a Markov model to compare omalizumab add-on therapy with standard therapy. Patients transitioned between symptom-free, day-to-day, and exacerbation states. Our model had a lifetime horizon in which 5-year omalizumab add-on therapy was followed by standard therapy. Preference-based utilities were extracted from another study. We estimated the expected value of perfect information for patients' response to omalizumab.ResultsIn the base case, incremental cost-effectiveness ratio (ICER) for omalizumab add-on therapy was US $755,200 (95$614,200–$1,298,500) per quality-adjusted life-year gained, compared with standard therapy alone. One-way sensitivity analyses indicated that the results were sensitive to asthma-related mortality, exacerbation risk, and omalizumab cost. The ICER for a responder subgroup was 22$4100 (95% CI $2500–$6000) and $28 million (95$17 million–$42 million) per year, respectively.ConclusionsWith a willingness-to-pay of$45,000 per quality-adjusted life-year, omalizumab was not cost-effective in Japan. Confining omalizumab therapy to previously predicted responders, however, may be a reasonable strategy to reduce the ICER, as the cost-effectiveness was observed to improve for these patients. Further studies should be conducted to explore responder prediction methods. Decreasing the price of omalizumab would improve cost-effectiveness

Effect of Ultraviolet Irradiation on Mast Cell-Deficient W/Wv Mice

The effect of UV irradiation on the skin was investigated in (WB-W/+) × (C57BL/6J-Wv/+)F1-W/Wv mice, which are genetically deficient in tissue mast cells. Their congenic littermates (+/+) and normal albino mice (ICR or Balb/c) were used as controls. Mice were irradiated with 500 mJ/cm2 of UVB and the increment of ear thickness was measured before and 6, 12, and 24 h after irradiation was significantly smaller than that in +/+ and ICR mice. In contrast, the number of sunburn cells formed 24 h after UVB irradiation (200 or 500 mJ/cm2) was similar in W/Wv, +/+ and ICR mice. On the other hand, when mice were treated with 8-methoxy-psoralen (0.5%) plus UVA irradiation (4 J/cm2) (topical PUVA), ears of W/Wv and BALB/c mice, which were both white in color, were thickened similarly 72 h after treatment, but less swelling was observed in +/+ mice, which were black in skin color. The amount of prostaglandin D2 (PGD2) in ears, determined by radioimmuno-assay specific for PGD2, was elevated 3-fold in +/+ and ICR mice at 3 h after irradiation with 500 mJ/cmv of UVB in comparison with basal level without irradiation. However, such elevation was not observed in W/Wv mice. These results suggest that mast cells play an important role in UVB-induced inflammation, and PGs from mast cells are responsible at lest in part for the development of this reaction. However, neither mast cells nor PGs contribute to the sunburn cell formation and ear swelling response by PUVA treatment

Two-Fingered Haptic Device for Robot Hand Teleoperation

A haptic feedback system is required to assist telerehabilitation with robot hand. The system should provide the reaction force measured in the robot hand to an operator. In this paper, we have developed a force feedback device that presents a reaction force to the distal segment of the operator's thumb, middle finger, and basipodite of the middle finger when the robot hand grasps an object. The device uses a shape memory alloy as an actuator, which affords a very compact, lightweight, and accurate device

mRNA detection of individual cells with the single cell nanoprobe method compared with in situ hybridization

<p>Abstract</p> <p>Background</p> <p>The localization of specific mRNA generates cell polarity by controlling the translation sites of specific proteins. Although most of these events depend on differences in gene expression, no method is available to examine time dependent gene expression of individual living cells. <it>In situ </it>hybridization (ISH) is a powerful and useful method for detecting the localization of mRNAs, but it does not allow a time dependent analysis of mRNA expression in single living cells because the cells have to be fixed for mRNA detection. To overcome these issues, the extraction of biomolecules such as mRNAs, proteins, and lipids from living cells should be performed without severe damage to the cells. In previous studies, we have reported a single cell nanoprobe (SCN) method to examine gene expression of individual living cells using atomic force microscopy (AFM) without killing the cells.</p> <p>Results</p> <p>In order to evaluate the SCN method, we compared the SCN method with <it>in situ </it>hybridization (ISH). First, we examined spatial β-actin mRNA expression in single living cells with the SCN method, and then the same cells were subjected to ISH for β-actin mRNA. In the SCN method, quantity of β-actin mRNA were analysed by quantitative PCR, and in ISH we used intensity of ISH as a parameter of concentration of β-actin mRNA. We showed that intensity of ISH is higher; quantity of β-actin mRNA detected by the SCN method increased more.</p> <p>Conclusion</p> <p>In this study, we compare the SCN method with the ISH. We examined β-actin mRNA expression in single cells using both methods. We picked up β-actin mRNA from several loci of a single living cell using an AFM nanoprobe, and identical cells were subjected to ISH. The results showed a good correlation between the SCN method and ISH. The SCN method is suitable and reliable to examine mRNAs at medium or higher expression level.</p

Self-organizing magnetic beads for biomedical applications

In the field of biomedicine magnetic beads are used for drug delivery and to treat hyperthermia. Here we propose to use self-organized bead structures to isolate circulating tumor cells using lab-on-chip technologies. Typically blood flows past microposts functionalized with antibodies for circulating tumor cells. Creating these microposts with interacting magnetic beads makes it possible to tune the geometry in size, position and shape. We developed a simulation tool that combines micromagnetics and discrete particle dynamics, in order to design micropost arrays made of interacting beads. The simulation takes into account the viscous drag of the blood flow, magnetostatic interactions between the magnetic beads and gradient forces from external aligned magnets. We developed a particle-particle particle-mesh method for effective computation of the magnetic force and torque acting on the particles

Jordan pairs of quadratic forms with values in invertible modules

Jordan pairs of quadratic forms are generalized so that they have forms with values in invertible modules. The role of such pairs turns out to be natural in describing ‘big cells’, a kind of open charts around unit sections, of Clifford and orthogonal groups as group schemes. Group germ structures on big cells are particularly interested in and related also to Cayley-Lipschitz transforms

Nutrient limitations alter cell division control and chromosome segregation through growth-related kinases and phosphatases

In dividing fission yeast Schizosaccharomyces pombe cells, the balance between Wee1 kinase and Cdc25 phosphatase which control the cyclin-dependent kinase (CDK) at the G2–M transition determines the rod-shaped cell length. Under nitrogen source starvation or glucose limitation, however, cell size determination is considerably modulated, and cell size shortening occurs for wild-type cells. For several mutants of kinases or phosphatases, including CDK, target of rapamycin complex (TORC) 1 and 2, stress-responsive mitogen-activated protein kinase (MAPK) Sty1/Spc1, MAPK kinase Wis1, calcium- and calmodulin-dependent protein kinase kinase-like Ssp1, and type 2A and 2A-related phosphatases inhibitor Sds23, this cell shortening does not normally occur. In tor1 and ssp1 mutants, cell elongation is observed. Sds23 that binds to and inhibits 2A and 2A-related phosphatases is synergistic with Ssp1 in the cell size determination and survival under low glucose and nitrogen source. Tor2 (TORC1) is required for growth, whereas Tor1 (TORC2) is needed for determining division size according to different nutrient conditions. Surprisingly, in growth-diminished tor2 mutant or rapamycin-treated cells, the requirement of separase/Cut1-securin/Cut2 essential for chromosome segregation is greatly alleviated. By contrast, defects of tor1 with secruin/cut2 or overproduction of Cut1 are additive. While Tor1 and Tor2 are opposite in their apparent functions, both may actually coordinate cell division with growth in response to the changes in nutrients