[Effective Techniques to Reduce Radiation Exposure to Medical Staff during Assist of X-ray Computed Tomography Examination].

Medical staffs like radiological technologists, doctors, and nurses are at an increased risk of exposure to radiation while assisting the patient in a position or monitor contrast medium injection during computed tomography (CT). However, methods to protect medical staff from radiation exposure and protocols for using radiological protection equipment have not been standardized and differ among hospitals. In this study, the distribution of scattered X-rays in a CT room was measured by placing electronic personal dosimeters in locations where medical staff stands beside the CT scanner gantry while assisting the patient and the exposure dose was measured. Moreover, we evaluated non-uniform exposure and revealed effective techniques to reduce the exposure dose to medical staff during CT. The dose of the scattered X-rays was the lowest at the gantry and at the examination table during both head and abdominal CT. The dose was the highest at the trunk of the upper body of the operator corresponding to a height of 130 cm during head CT and at the head corresponding to a height of 150 cm during abdominal CT. The maximum dose to the crystalline lens was approximately 600 µSv during head CT. We found that the use of volumetric CT scanning and X-ray protective goggles, and face direction toward the gantry reduced the exposure dose, particularly to the crystalline lens, for which lower equivalent dose during CT scan has been recently recommended in the International Commission on Radiological Protection Publication 118.

DNA-duplex linker for AFM-SELEX of DNA aptamer against human serum albumin.

DNA-duplex interactions in thymines and adenins are used as a linker for the novel methodology of Atomic Force Microscope-Systematic Evolution of Ligands by EXpotential enrichment (AFM-SELEX). This study used the hydrogen bonds in 10 mer of both thymines (T10) and adenines (A10). Initially, the interactive force in T10-A10 was measured by AFM, which returned an average interactive force of approximately 350pN. Based on this result, DNA aptamers against human serum albumin could be selected in the 4th round, and 15 different clones could be sequenced. The lowest dissociation constant of the selected aptamer was identified via surface plasmon resonance, and it proved to be identical to that of the commercial aptamer. Therefore, specific hydrogen bonds in DNA can be useful linkers for AFM-SELEX.

Cell-SELEX based selection and characterization of DNA aptamer recognizing human hepatocarcinoma.

Single-stranded DNA aptamers recognizing human hepatocarcinoma were isolated by means of a systematic evolution of ligands by exponential enrichment using whole cells as targets (cell-SELEX). After 11 rounds of cell-SELEX procedure using human hepatoma HepG2 cells as targets and human normal hepatocyte cells as counterparts, 12 independent DNA aptamer candidate sequences were obtained. The specific interaction between selected DNA aptamers and targeted cell was confirmed. Dissociation constants of the 12 sequences obtained were also estimated in the range of 19-450nM. Moreover, the consensus secondary structure was found in the isolated aptamers, which was responsible to the recognition of HepG2 cells.

Selection of DNA aptamers using atomic force microscopy.

Atomic force microscopy (AFM) can detect the adhesion or affinity force between a sample surface and cantilever, dynamically. This feature is useful as a method for the selection of aptamers that bind to their targets with very high affinity. Therefore, we propose the Systematic Evolution of Ligands by an EXponential enrichment (SELEX) method using AFM to obtain aptamers that have a strong affinity for target molecules. In this study, thrombin was chosen as the target molecule, and an 'AFM-SELEX' cycle was performed. As a result, selected cycles were completed with only three rounds, and many of the obtained aptamers had a higher affinity to thrombin than the conventional thrombin aptamer. Moreover, one type of obtained aptamer had a high affinity to thrombin as well as the anti-thrombin antibody. AFM-SELEX is, therefore, considered to be an available method for the selection of DNA aptamers that have a high affinity for their target molecules.

Construction of protein-modified TiO2 nanoparticles for use with ultrasound irradiation in a novel cell injuring method.

Recently, our group discovered an alternative titanium dioxide (TiO(2)) activation method that uses ultrasound irradiation (US/TiO(2)) instead of ultraviolet irradiation. The pre-S1/S2 protein from hepatitis B virus, which recognizes liver cells, was immobilized to the surface of TiO(2) nanoparticles using an amino-coupling method. The ability of the protein-modified TiO(2) nanoparticles to recognize liver cells was confirmed by surface plasmon resonance analysis and immuno-staining analyses. After uptake of TiO(2) nanoparticles by HepG2 cancer cells, the cells were injured using this US/TiO(2) method; significant cell injury was observed at an ultrasound irradiation intensity of 0.4 W/cm(2). Together with these results, this strategy could be applied to new cell injuring systems that use ultrasound irradiation in place of photodynamic therapy in the near future.

Selection of a DNA aptamer that binds 8-OHdG using GMP-agarose.

DNA aptamers, which bind specific molecule, such as 8-OHdG, with high affinity were investigated using an in vitro selection strategy called systematic evolution of ligands by exponential enrichment (SELEX). However, 8-OHdG was difficult to immobilize on a carrier for SELEX. Therefore, a DNA aptamer binding to 8-OHdG was selected using GMP-agarose as an analogue from a library of about 4(60) random ssDNA sources. As a result, three aptamer candidates were selected. Among the selected DNA aptamers, the No. 22 DNA aptamer exhibited a high affinity for 8-OHdG. The dissociation constant, K(D), of No. 22 DNA aptamer was on the order of 0.1micromol/L. This result suggests that using an analogue will be a useful new SELEX method for obtaining various aptamers that are difficult to immobilize on a matrix.

The mapping of yeast's G-protein coupled receptor with an atomic force microscope.

An atomic force microscope (AFM) can measure the adhesion force between a sample and a cantilever while simultaneously applying a rupture force during the imaging of a sample. An AFM should be useful in targeting specific proteins on a cell surface. The present study proposes the use of an AFM to measure the adhesion force between targeting receptors and their ligands, and to map the targeting receptors. In this study, Ste2p, one of the G protein-coupled receptors (GPCRs), was chosen as the target receptor. The specific force between Ste2p on a yeast cell surface and a cantilever modified with its ligand, α-factor, was measured and found to be approximately 250 pN. In addition, through continuous measuring of the cell surface, a mapping of the receptors on the cell surface could be performed, which indicated the differences in the Ste2p expression levels. Therefore, the proposed AFM system is accurate for cell diagnosis.

Structural evaluation of the DNA aptamer for ATP DH25.42 by AFM.

Atomic force microscopy (AFM) can dynamically detect the adhesion or affinity force between a sample surface and a cantilever. This feature could be used to analyze bio-molecular interactions between a DNA aptamer and a target molecule. In this study, the binding force between adenosine triphosphate (ATP) and the anti-ATP DNA aptamer DH25.42, based on structural changes was measured using AFM. In addition, the relationship between the cations in the binding buffer, and the affinity and structure formation of the DNA aptamer was also evaluated using AFM and circular dichroism (CD) spectrum analysis. As a result, the specific force between DH25.42 and ATP could be measured by AFM. Moreover, it was suggested that Mg(2+) in the binding buffer was critical to the binding function of DH25.42 to ATP.

Development of a novel aptamer-based sensing system using atomic force microscopy.

Atomic force microscopy (AFM) can dynamically detect the adhesion or affinity force between a sample surface and a cantilever. This feature is useful as a detection method using aptamers--single-strand DNA that recognizes its target with very high affinity. The present study proposes a novel DNA aptamer-based sensing system using AFM. In this study, thrombin was chosen as the target molecule, and a DNA aptamer-based AFM sensing system based on competition was developed. The affinity force between the gold chip and the cantilever decreased as the concentration of thrombin increased. Moreover, a low detection limit of 0.2 nM was achieved. Therefore, the AFM sensing system used would be appropriate for the measurement of various chemical compounds.