Analysis of individual differences in radiosensitivity using genome editing.

Current standards for radiological protection of the public have been uniformly established. However, individual differences in radiosensitivity are suggested to exist in human populations, which could be caused by nucleotide variants of DNA repair genes. In order to verify if such genetic variants are responsible for individual differences in radiosensitivity, they could be introduced into cultured human cells for evaluation. This strategy would make it possible to analyse the effect of candidate nucleotide variants on individual radiosensitivity, independent of the diverse genetic background. However, efficient gene targeting in cultured human cells is difficult due to the low frequency of homologous recombination (HR) repair. The development of artificial nucleases has enabled efficient HR-mediated genome editing to be performed in cultured human cells. A novel genome editing strategy, 'transcription activator-like effector nuclease (TALEN)-mediated two-step single base pair editing', has been developed, and this was used to introduce a nucleotide variant associated with a chromosomal instability syndrome bi-allelically into cultured human cells to demonstrate that it is the causative mutation. It is proposed that this editing technique will be useful to investigate individual radiosensitivity.

Minimally required heat doses for various tumour sizes in induction heating cancer therapy determined by computer simulation using experimental data.

PURPOSE: Although induction heating cancer therapy (IHCT) using magnetic nanoparticles can be a promising approach to treatment-less multi-nodular cancers, the objective requirement for successful clinical application has not clearly been elucidated. We intended to define objective heat doses suitable for IHCT, especially focusing on the sizes of liver cancer nodules. MATERIALS AND METHODS: Alternating magnetic fields were applied to three human pancreatic cancer cell lines, the intercellular space of those cell pellets were filled with magnetic nanoparticles, and confirmed the cytotoxic effect of IHCT. Subsequently, the temperatures of liver cancer nodules in IHCT were simulated using a computer software program and the required heat dose for various sized tumours were determined. RESULTS: Heating the cancer cells up to 50 degrees C for 10 min was sufficient for complete cell killing and the heat dose of 1.7 W/g(tumour) is required for 10 mm tumour. Larger tumours require a smaller heat dose, e.g. 20 mm and 40 mm tumours require 0.7 W/g(tumour) and 0.6 W/g(tumour), respectively, whereas smaller tumours require large amounts of heat, e.g. 5 mm and 1 mm tumours require 5.1 W/g(tumour) and 105 W/g(tumour), respectively. CONCLUSIONS: Integrating the presently available technologies, including high-quality magnetic nanoparticles (1000 W/g(material)) and effective drug delivery systems (1-2 mg(material)/g(tumour)), treatment of a 10 mm tumour seems possible. Since treatment of smaller tumours less than 5 mm require substantial heat dose, researchers involved in IHCT should target cancer nodules of 10 mm or more, and develop a heat delivery system providing a minimum of 1.7 W/g(tumour).

The measurement of small magnetic signals from magnetic nanoparticles attached to the cell surface and surrounding living cells using a general-purpose SQUID magnetometer.

Magnetic nanoparticles have recently been widely applied in the bio-medical field. Responding to the demand for a simple and sensitive magnetic assay system for bio-liquid samples, we employed a general-purpose superconducting quantum interference device (SQUID) magnetometer. Strips of filter paper were used as a liquid-specimen sample holder possessing a very small magnetic background signal. An aqueous solution of superparamagnetic iron-oxide nanoparticles (Resovist) was dropped in a tiny blot-like spot in the middle of the filter paper and the magnetization was measured. Magnetic moments of a dilution series of Resovist solutions versus the number of particles provided a linear graph, revealing that the magnetic moment per Resovist particle was 8.25 x 10(-17) emu. 1 x 10(5) cancer cells were incubated with Resovist, and the number of Resovist particles attached to the cell surface and surrounding a living cell was calculated to be 1.02 +/- 0.14 x 10(7) particles/cell. Our system using a commercial SQUID magnetometer should be more than enough to determine the number of magnetic nanoparticles biologically reacting with living cells, contributing to the application of magneto nanomaterials to the life-science field.

Human CYP3A4-introduced HepG2 cells: in vitro screening system of new chemicals for the evaluation of CYP3A4-inhibiting activity.

1. The aims were to attest whether HepG2-GS-3A4, a cell line into which the human CYP3A4 gene was introduced, can be used for a screening of chemicals that will inhibit CYP3A4 activity. 2. The capacity of the cells for metabolizing CYP3A4 substrates in vitro was evaluated. Also determined was the effect of CYP3A4 inhibitors and non-inhibitors on nifedipine hydroxylation. Western blot, immunohistochemostry and determination of beta-nicotinamide adenine dinucleotide phosphate (NADPH)-reductase activity were performed. 3. HepG2-GS-3A4 selectively metabolized substrates of CYP3A4 (diazepam, nordiazepam, lidocaine, atorvastatin, and nifedipine) to a greater degree than control. The metabolites were easily detected in the culture medium. Values of V(max) of HepG2-GS-3A4 were about 30- to 100-fold higher than those of the control, while values of K(m) were comparable. Pre-incubation of cimetidine and ketoconazole significantly inhibited nifedipine hydroxylation, while addition of inhibitors specific to other isoforms of CYPs had no substantial effect. The HepG2-GS-3A4 expressed a higher amount of CYP3A4 protein and mRNA than control. Most NADPH reductase activity was detected in microsomal fractions. 4 In conclusion, HepG2-GS-3A4 sufficiently and selectively metabolize substrates of CYP3A4, and inhibitors of CYP3A4 reduced the metabolism. Because the metabolites were easily detected in the culture medium, this cell might be useful for the new and easy screening of new drugs for the evaluation of CYP3A4-inhibiting activity in vitro.

The role of topological spin defects in magnetotransport of CrO(2).

We investigated the temperature dependence of the resistivity for a wide temperature range for CrO(2)(100) epitaxial films. The temperature derivative dρ/dT definitely shows the same character as the magnetic heat capacity anomaly in the critical regime even in a finite magnetic field and the critical exponents (α) deduced are consistent with those of 3D Heisenberg ferromagnets. In addition, we found that the spin dependent resistivity over a wide temperature range can be simply proportional to the density of diluted topological spin defects (Skyrmion strings) suggesting that those nontrivial topological defects scatter conduction electrons just like impurities. The excitation energy of such topological defects is quite comparable to that obtained by anomalous Hall effect analysis of the Ye et al model based on the Berry phase. The overall results give a simple picture wherein the density of the topological defects can be a dominant mechanism of resistivity, like the anomalous Hall effect. The results concerning the critical exponent analysis and intuition concerning scattering centres of magnetic disorder suggest a specific picture of the Fisher-Langer model.

Dynamics of water droplets detached from porous surfaces of relevance to PEM fuel cells.

The detachment of liquid droplets from porous material surfaces used with proton exchange membrane (PEM) fuel cells under the influence of a cross-flowing air is investigated computationally and experimentally. CCD images taken on a purpose-built transparent fuel cell have revealed that the water produced within the PEM is forming droplets on the surface of the gas-diffusion layer. These droplets are swept away if the velocity of the flowing air is above a critical value for a given droplet size. Static and dynamic contact angle measurements for three different carbon gas-diffusion layer materials obtained inside a transparent air-channel test model have been used as input to the numerical model; the latter is based on a Navier-Stokes equations flow solver incorporating the volume of fluid (VOF) two-phase flow methodology. Variable contact angle values around the gas-liquid-solid contact-line as well as their dynamic change during the droplet shape deformation process, have allowed estimation of the adhesion force between the liquid droplet and the solid surface and successful prediction of the separation line at which droplets loose their contact from the solid surface under the influence of the air stream flowing around them. Parametric studies highlight the relevant importance of various factors affecting the detachment of the liquid droplets from the solid surface.

In vivo assessment of the cutaneous bioavailability of topically applied maxacalcitol.

Maxacalcitol (22-oxacalcitriol), a vitamin D3 analogue, is widely used for the treatment of psoriasis. The effects of topical dermatologic drugs have been assessed by their pharmacodynamic activities, and concentrations in the skin correlate with these activities. In this study, we assessed the cutaneous bioavailability of topically applied maxacalcitol ointment in vivo by tape stripping. Six drug application sites were randomly assigned on the left volar forearm of six healthy men. Fifty milligrams of maxacalcitol ointment (25 microg/g) was applied to each site. After 0 (15 s), 0.5, 1, 2, 4, or 6 h, the ointment was gently removed, and tape stripping was performed. Maxacalcitol was extracted from the tape strips and quantified by liquid chromatographic tandem mass spectrometry. Average concentrations of maxacalcitol in the stratum corneum (SC) were 2.61, 4.37, 6.23, 9.37, and 9.46 microg/g at 0.5, 1, 2, 4, and 6 h, respectively, after drug application. The steady state was attained approx. 4 h after drug application. The cutaneous bioavailability of topical maxacalcitol ointment can be assessed by the tape-stripping method. This approach will probably be useful in the assessment of the bioequivalence of topical dermatologic products and as a parameter for pharmacokinetic/pharmacodynamic studies.

Skyrmion strings and the anomalous Hall effect in CrO2.

Topological (or singularity point) defects are thought to play a crucial role in the phase transitions of 3D spin systems, as they do in such 2D systems as the XY model. In double-exchange ferromagnets the conduction electrons are strongly coupled with core spins through Hund's rule, and, in the presence of a nontrivial spin texture, acquire a Berry phase contribution to the anomalous Hall effect. We combine Hall effect and magnetization data on CrO2 with a thermodynamical scaling hypothesis to confirm that the critical behavior of the topological-spin-defect density is consistent with that of the heat capacity. This analysis is the first experimental confirmation of the topological character of critical fluctuations.

[Osteoma originating in the dura: a case report].

We report a case of intracranial osteoma attached to the dura. The patient, a 35-year-old man, had suffered several episodes of vertigo over the previous two years. Physical and neurological findings on admission were unremarkable. A plain craniogram showed a dense calcified mass in the right frontal area, and CT revealed a homogeneous high-density mass without significant enhancement. MRI confirmed the dural origin of the lesion, which showed hyperintensity in the T1-weighted image and low intensity in the T2-weighted image. The mass was 5 cm in diameter and 2 cm in thickness. Total resection was performed under a preoperative diagnosis of calcified meningioma. Histopathology revealed the mass to be an osteoma. Osteomas are common benign bone tumors that usually arise from long bones of the extremities. Intracranial osteomas are extremely rare lesions. The literature on intracranial osteoma of dural origin is reviewed.

Rapid analysis of small samples containing forskolin using monoclonal antibodies.

The effective range of the competitive ELISA test for detection of forskolin content in clonally propagated plant organs of Coleus forskohlii using monoclonal antibodies extends from 5ng to 5 micrograms. A correlation between the forskolin accumulation and the growth rate was investigated using the clonally propagated shoots. An increase of forskolin content was noted, beginning at week 6. Flowers, rachises, leaves, stems, tuberous roots, and roots were analyzed. Tuberous roots and the stem base contained higher amounts of forskolin than other organs. The forskolin content in the stem decreased gradually towards the top of the shoot.

Experimental study on autograft material for repair of round window rupture.

The healing process following experimentally produced rupture of the round window (RW) and subsequent closure with autograft materials were investigated in guinea pigs. The covering materials studied were fascia and perichondrium, both of which were placed over the RW after extensive rupturing of the RW membrane. The grafts were observed 1, 2, and 3 months after the operation. No substantial difference in the results for the two materials could be discerned. They adhered to the surrounding bone and were incorporated into the RW membrane within 3 months.

Goldenhar's syndrome associated with anomalous internal auditory meatus.

A case of Goldenhar's syndrome associated with fifth, seventh, and eighth nerve involvement is described. Tomographic examination of the temporal bones showed evidence of an abnormal internal auditory meatus. Otoneurological examination suggested that involvement of the seventh and eighth nerves was closely related to the abnormality in the internal auditory meatus.