Hyperglycemia Induces Skin Barrier Dysfunctions with Impairment of Epidermal Integrity in Non-Wounded Skin of Type 1 Diabetic Mice.

Diabetes causes skin complications, including xerosis and foot ulcers. Ulcers complicated by infections exacerbate skin conditions, and in severe cases, limb/toe amputations are required to prevent the development of sepsis. Here, we hypothesize that hyperglycemia induces skin barrier dysfunction with alterations of epidermal integrity. The effects of hyperglycemia on the epidermis were examined in streptozotocin-induced diabetic mice with/without insulin therapy. The results showed that dye leakages were prominent, and transepidermal water loss after tape stripping was exacerbated in diabetic mice. These data indicate that hyperglycemia impaired skin barrier functions. Additionally, the distribution of the protein associated with the tight junction structure, tight junction protein-1 (ZO-1), was characterized by diffuse and significantly wider expression in the diabetic mice compared to that in the control mice. In turn, epidermal cell number was significantly reduced and basal cells were irregularly aligned with ultrastructural alterations in diabetic mice. In contrast, the number of corneocytes, namely, denucleated and terminally differentiated keratinocytes significantly increased, while their sensitivity to mechanical stress was enhanced in the diabetic mice. We found that cell proliferation was significantly decreased, while apoptotic cells were comparable in the skin of diabetic mice, compared to those in the control mice. In the epidermis, Keratin 5 and keratin 14 expressions were reduced, while keratin 10 and loricrin were ectopically induced in diabetic mice. These data suggest that hyperglycemia altered keratinocyte proliferation/differentiation. Finally, these phenotypes observed in diabetic mice were mitigated by insulin treatment. Reduction in basal cell number and perturbation of the proliferation/differentiation process could be the underlying mechanisms for impaired skin barrier functions in diabetic mice.

[Clinical Application of Cancer Molecular Diagnostics with Liquid Biopsy].

"Liquid biopsy" is one of the hottest topics in the cancer molecular diagnostics field because of its potential to replace conventional tissue biopsy. "BEAMing" is one of the digital PCR platform technologies for liquid biopsy samples, and it features a high enough sensitivity to detect a very low concentration of mutation frag- ments in circulating tumor DNA from the plasma of cancer patients. In this article, some of the promising clinical applications of BEAMing technology and its future possibilities as an advanced cancer diagnostic will be briefly summarized. [Review].

Reconsideration of the Iwasaki-Waggener iterative perturbation method for reconstructing high-energy X-ray spectra.

We have reviewed applicable ranges for attenuating media and off-axis distances regarding the high-energy X-ray spectra reconstructed via the Iwasaki-Waggener iterative perturbation method for 4-20 MV X-ray beams. Sets of in-air relative transmission data used for reconstruction of spectra were calculated for low- and high-Z attenuators (acrylic and lead, respectively) by use of a functional spectral formula. More accurate sets of spectra could be reconstructed by dividing the off-axis distances of R = 0-20 cm into two series of R = 0-10 cm and R = 10-20 cm, and by taking into account the radiation attenuation and scatter in the buildup cap of the dosimeter. We also incorporated in the reconstructed spectra an adjustment factor (f (adjust) ≈ 1) that is determined by the attenuating medium, the acceleration voltage, and the set of off-axis distances. This resulted in calculated in-air relative transmission data to within ±2 % deviation for the low-Z attenuators water, acrylic, and aluminum (Al) with 0-50 cm thicknesses and R = 0-20 cm; data to within ±3 % deviation were obtained for high-Z attenuators such as iron (Fe), copper (Cu), silver (Ag), tungsten (W), platinum (Pt), gold (Au), lead (Pb), thorium (Th), and uranium (U) having thicknesses of 0-10 cm and R = 0-20 cm. By taking into account the radiation attenuation and scatter in the buildup cap, we could analyze the in-air chamber response along a line perpendicular to the isocenter axis.

A convolution/superposition method using primary and scatter dose kernels formed for energy bins of X-ray spectra reconstructed as a function of off-axis distance: comparison of calculated and measured 10-MV X-ray doses in thorax-like phantoms.

We performed experimental studies on the convolution/superposition method reported in the former companion paper (Iwasaki in Radiol Phys Technol 4, 2011) using 10-MV X-ray beams from open-jaw-collimated fields. The method uses primary and scatter dose kernels formed for energy bins of X-ray spectra reconstructed as a function of off-axis distance. We made a comparison of calculations and measurements in water phantoms and thorax-like phantoms with respect to percentage depth dose curves, tissue-phantom ratio curves, and dose profiles. We made the dose calculation by taking into account the beam-hardening effect with depth and the off-axis radiation-softening effect. We found that the method could be used, in general, for performing accurate dose calculations.

A convolution/superposition method using primary and scatter dose kernels formed for energy bins of X-ray spectra reconstructed as a function of off-axis distance: a theoretical study on 10-MV X-ray dose calculations in thorax-like phantoms.

A convolution/superposition method is proposed for use with primary and scatter dose kernels formed for energy bins of X-ray spectra reconstructed as a function of off-axis distance. It should be noted that the number of energy bins is usually about ten, and that the reconstructed X-ray spectra can reasonably be applied to media with a wide range of effective Z numbers, ranging from water to lead. The study was carried out for 10-MV X-ray doses in water and thorax-like phantoms with the use of open-jaw-collimated fields. The dose calculations were made separately for primary, scatter, and electron contamination dose components, for which we used two extended radiation sources: one was on the X-ray target and the other on the flattening filter. To calculate the in-air beam intensities at points on the isocenter plane for a given jaw-collimated field, we introduced an in-air output factor (OPF(in-air)) expressed as the product of the off-center jaw-collimator scatter factor (off-center S (c)), the source off-center ratio factor (OCR(source)), and the jaw-collimator radiation reflection factor (RRF(c)). For more accurate dose calculations, we introduce an electron spread fluctuation factor (F (fwd)) to take into account the angular and spatial spread fluctuation for electrons traveling through different media.

Characteristic features of a high-energy x-ray spectra estimation method based on the Waggener iterative perturbation principle.

We have redeveloped a high-energy x-ray spectra estimation method reported by Iwasaki et al. [A. Iwasaki, H. Matsutani, M. Kubota, A. Fujimori, K. Suzaki, and Y. Abe, Radiat. Phys. Chem. 67, 81-91 (2003)]. The method is based on the iterative perturbation principle to minimize differences between measured and calculated transmission curves, originally proposed by Waggener et al. [R. G. Waggener, M. M. Blough, J. A. Terry, D. Chen, N. E. Lee, S. Zhang, and W. D. McDavid, Med. Phys. 26, 1269-1278 (1999)]. The method can estimate spectra applicable for media at least from water to lead using only about ten energy bins. Estimating spectra of 4-15 MV x-ray beams from a linear accelerator, we describe characteristic features of the method with regard to parameters including the prespectrum, number of transmission measurements, number of energy bins, energy bin widths, and artifactual bipeaked spectrum production.

Theoretical considerations for evaluating the degree of random-periodicity of radiographic noise.

Radiographic noise properties have been evaluated using the Wiener spectrum. However, this approach is not appropriate for periodic noise for two reasons. One is that it takes infinite values at the spatial frequencies of periodic noise. The other is that when adopting a numerical integration, it allows unstable values at each spatial frequency, depending on the integral region. Introducing three types of spectra (W(1), W(2), and W(3)) in connection with the Wiener spectrum, we propose a practical approach to evaluation of periodic noise. Radiographic images sometimes contain noise that is not totally random and not perfectly periodic. Therefore, using the Wiener spectrum and the W(1) spectrum, we also propose two factors for evaluation of the degree of random-periodicity of noise containing periodic signals.

[Theoretical considerations for evaluating the degree of random-periodicity of graphic noise.].

Graphic noise properties are usually evaluated using the Wiener spectrum. However, this approach is not appropriate for periodic noise for two reasons. One is that it takes infinite values at the spatial frequencies of periodic noise. The other is that when adopting a numerical integration, it allows unstable values at each spatial frequency, depending on the integral region. Introducing three types of spectra (W(1), W(2), and W(3)) in connection with the Wiener spectrum, we propose a practical approach to evaluation of periodic noise. Graphic images sometimes contain noise that is not totally random and not perfectly periodic. Therefore, using the Wiener spectrum and the W(1) spectrum, we also propose two factors for evaluation of the degree of random-periodicity of noise containing periodic signals.

Establishment of Periodontal Ligament Cell Lines from Temperature-Sensitive Simian Virus 40 Large T-antigen Transgenic Rats.

Orthodontic tooth movement is controlled by various cell types in the periodontal ligament (PDL). Mechanical stresses, such as orthodontic force, are thought to induce differentiation of the mesenchymal cells in the PDL into osteoblasts and cementoblasts. The details of the process of differentiation, however, are not known, in part because adequate in vitro systems for their study do not yet exist. The purpose of this study was to establish and characterize immortalized PDL cell lines derived from the PDL of transgenic rats harboring the temperature-sensitive simian virus 40 T-antigen gene (TG rats). The PDL was removed from the molar roots of TG rats and incubated in tissue culture. Outgrowth cells from the PDL explant were passaged and cloned, depending on the shape of the colonies formed. The cell lines thus established were analyzed by reverse transcription-polymerase chain reaction for expression of type-I collagen, osteopontin, fibronectin, alkaline phosphatase (bone type), bone sialoprotein, the receptor activator of NF-kappa B ligand, and osteoprotegerin. In addition, the capacity for formation of mineralized nodules was assessed by incubating cells in calcification-promoting medium at 37 degrees C. A total of 15 stable cell lines were successfully established and characterized. These cell lines were classified into six groups based on their pattern of gene expression at 33 degrees C. Moreover, three of these clones were capable of forming calcified nodules. In conclusion, differential gene expression was demonstrated in 15 established PDL cell lines. Some cells had the potential to differentiate into cell types found in mineralized tissues, such as osteoblasts and cementoblasts, as well as cells expressing molecules that regulate osteoclast differentiation.

Novel enzyme-linked immunosorbent assay systems for the quantitative analysis of connective tissue growth factor (CTGF/Hcs24/CCN2): detection of HTLV-I tax-induced CTGF from a human carcinoma cell line.

Connective tissue growth factor/hypertrophic chondrocyte-specific gene product 24 (CTGF/Hcs24/CCN2) is known as a multifunctional growth factor. It stimulates proliferation, migration, and extracellular matrix production of mesenchymal cells, and is highly expressed in hypertrophic chondrocytes. In this study, we constructed useful ELISA systems for the analysis of CTGF and its modular fragments. For this objective we prepared four different antihuman CTGF monoclonal antibodies. One, specific for the VWC module, was utilized as the detecting antibody, and the other three, recognizing CT, IGFBP, and VWC modules, respectively, were employed as capture antibodies. Then we established three novel quantitative analysis systems for CTGF. The first system recognizing CT and VWC modules was useful to measure full-length CTGF with improved sensitivity. Utilizing this system, we found significant enhancement of CTGF production from a human carcinoma cell line transduced by HTLV-I tax gene, where the finding indicates the possible involvement of Tax in carcinogenesis. The second system, seeing IGFBP and VWC modules, could quantify not only CTGF, but also may be useful to analyze processed N-terminal fragments. The third system, utilizing capture and detection antibodies against the VWC module, was able to quantify the VWC module only, while it did not recognize full-length CTGF. Since CTGF is actually processed into subfragments, and functional assignment of each module is of interest, these systems are expected to contribute to the progress of CTGF investigations.