Possible Involvement of the Upregulation of ΔNp63 Expression Mediated by HER2-Activated Aryl Hydrocarbon Receptor in Mammosphere Maintenance

Cancer stem cells (CSCs) contribute to the drug resistance, recurrence, and metastasis of breast cancers. Recently, we demonstrated that HER2 overexpression increases mammosphere formation via the activation of aryl hydrocarbon receptor (AHR). In this study, the objective was to identify the mechanism underlying mammosphere maintenance mediated by HER2 signaling-activated AHR. We compared the chromatin structure of AHR-knockout (AHRKO) HER2-overexpressing MCF-7 (HER2-5) cells with that of wild-type HER2-5 cells; subsequently, we identified TP63, a stemness factor, as a potential target gene of AHR. ΔNp63 mRNA and protein levels were higher in HER2-5 cells than in HER2-5/AHRKO cells. Activation of HER2/HER3 signaling by heregulin treatment increased ΔNp63 mRNA levels, and its induction was decreased by AHR knockdown in HER2-5 cells. The results of the chromatin immunoprecipitation assay revealed an interaction between AHR and the intronic region of TP63, which encodes ΔNp63. A luciferase reporter gene assay with the intronic region of TP63 showed that AHR expression increased reporter activity. Collectively, our findings suggest that HER2-activated AHR upregulates ΔNp63 expression and that this signaling cascade is involved in CSC maintenance in HER2-expressing breast cancers.     

Immunomagnetic flow cytometric detection of staphylococcal enterotoxin B in raw and dry milk

A rapid and sensitive method for detection of staphylococcal enterotoxin B (SEB) in raw and dry milk samples with the use of antibody-based immunomagnetic separation (IMS) in conjunction with flow cytometry (FCM) was developed. Sheep anti-SEB immunoglobulin G (IgG) was immobilized on Dynabeads M-280. The SEB initially binds to the capturing antibody, which is bound on the magnetic beads. The rabbit anti-SEB IgG binds to the captured toxin and is further labeled with a Cy5-labeled goat anti-rabbit IgG antibody. The percentage of the beads that were fluorescent was measured by FCM. FCM was carried out for 1 min, and the data obtained were expressed as histograms for particle size (forward light scatter) and histograms for fluorescence intensity. A peak corresponding to the magnetic beads was clearly distinguished from a peak derived from contaminating particles in the sample solution. In the absence of SEB, about 10% of the beads emitted fluorescence. The percentage of fluorescent beads and the fluorescence intensity increased with increasing SEB concentrations. For this IMS-FCM assay, the lower limits of detection for SEB were estimated to be 0.01 and 0.25 ng/ml for buffer and milk samples, respectively.     

Evaluation of drug toxicity with hepatocytes cultured in a micro-space cell culture system

A micro-space cell culture system was recently developed in which cells such as hepatocytes can be cultured and formed into a multicellular three-dimensional (3D) architecture. In this study, we assessed the performance of HepG2 cells cultured in this micro-space cell culture system in a drug toxicity test, and evaluated the effects of micro-space culture on their hepatocyte-specific functions. The micro-space cell culture facilitated the formation of 3D HepG2 cell architecture. HepG2 cells cultured in a micro-space culture plate exhibited increased albumin secretion and enhanced mRNA expression levels of cytochrome P450 (CYP) enzyme compared to those cultured in a monolayer culture. When the cells were exposed to acetaminophen, a hepatotoxic drug, the damage to the HepG2 cells grown in micro-space culture was greater than the damage to the HepG2 cells grown in monolayer culture. In addition, human primary hepatocytes grown in micro-space culture also exhibited increased albumin secretion, enhanced CYP mRNA expression levels and increased sensitivity to acetaminophen compared to those grown in monolayer culture. These results suggest that this micro-space culture method enhances the hepatocyte-specific functions of hepatocytes, including drug-metabolizing enzyme activities, making hepatocytes grown in the micro-space culture system a useful tool for evaluating drug toxicity in vitro.     

Grain Refinement by Cyclic Displacive Forward/Reverse Transformation in Fe-High-Ni Alloys

A novel method for grain refinement of martensite structures was proposed, in which transformation strain is accumulated by cyclic displacive forward and reverse transformations. This method can refine martensite structures in an Fe-18Ni alloy because a high density of austenite dislocations is introduced by a displacive reverse transformation in addition to an inheritance of dislocations in body-centered cubic martensite into austenite during cyclic transformation. The addition of a small amount of carbon accelerates structure refinement significantly, which results in the formation of ultra-fine-grained structures after ten cycles.

     

Effect of carbon content on variant pairing of martensite in Fe–C alloys

The effect of carbon content on the variant pairing tendency of martensite formed in Fe–C alloys is investigated by means of electron backscattered diffraction analysis. The method used is based on experimentally determined orientation relationships between austenite and martensite. The results show that the carbon content has a strong effect on the martensite variant pairing tendency. This observed change in variant pairing tendency is discussed in relation to the well-known morphological transition from lath to plate martensite in Fe–C alloys and the formation of packets and plate groups. The results indicate that quantitative analysis of variant pairing, as demonstrated here, may facilitate martensite characterization in Fe–C alloys as well as in other alloy systems.     

Wind Force Coefficients for Designing Porous Canopy Roofs

Wind force coefficients for designing porous canopy roofs have been investigated based on a series of wind tunnel experiments. Gable, troughed and mono-sloped roofs were tested. The roof models were made of 0.5 mm thick perforated duralumin plates, the porosity of which was changed from 0 to about 0.4. Overall aerodynamic forces and moments acting on the roof model were measured in a turbulent boundary layer with a six-component force balance for various wind directions. The results indicate that the wind loads on canopy roofs generally decrease with an increase in porosity of the roof. Assuming that the roof is rigid and supported by the four corner columns with no walls, the axial forces induced in the columns are regarded as the most important load effect for discussing the design wind loads. Two loading patterns causing the maximum tension and compression in the columns are considered. Based on a combination of the lift and moment coefficients, the design wind force coefficients on the windward and leeward halves of the roof are presented for the two loading patterns as a function of the roof pitch and porosity. The effect of porosity is taken into account as a reduction factor of the wind loads.     

Rotary scanning wireless temperature measurement method for hyperthermia using ferromagnetic implants

Recently, the use of a ferromagnetic material in a soft‐heating method has garnered much attention as a novel method for cancer treatment. By concurrently using this material as a thermal probe, we are currently developing a minimally invasive heating and wireless temperature measurement system. To make the approach feasible in a clinical setting, it is vital to overcome the key challenge of heating the local tumor at a constant temperature. In previous conventional approaches, it was necessary to switch the induction‐heating power supply on/off after the target tumor temperature was reached. However, it cannot determine the temperature of the material during the power‐off period. Therefore, we changed this approach and found that by adjusting the distance between the heating coil and the material while maintaining a constant current flow in heating coil, the drift problem, which happened just after power is supplied during the on/off operation, did not occur any longer. Accordingly, it was not required to use multiple sensors to reduce the drift, thereby minimizing the cost. This study verifies the validity of our wireless thermometry approach while performing rotary scanning and proposes a technique for determining achievement of the target temperature. This knowledge complements other approaches for cancer treatment utilizing hyperthermia. © 2015 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc.     

Surface Analysis of Cr2O3‐, CoO‐, and Al2O3‐Doped Iron–Phosphate Glasses at High Temperature

Surface structures of iron–phosphate glasses were examined using X‐ray photoelectron spectroscopy (XPS). Cr₂O₃, CoO, and Al₂O₃ were introduced to the glass by the replacement of a part of Fe₂O₃, and the simulated fission products are also added. The obtained glasses showed high chemical durabilities by MCC‐1 test. In situ high‐temperature and room‐temperature XPS measurements were conducted on the polished sample surfaces and also those after 1‐week chemical durability test. Unique trends were observed in XPS spectra on heating and after the chemical durability test, respectively. Nature of the glass surface of iron–phosphate glasses was explained from the point of view of surface energy, and the origin of high chemical durability and the effect of chromium ions were discussed based on the changes on surface composition and valence states of transition‐metal ions.     

Direct measurement of carbon enrichment during austenite to ferrite transformation in hypoeutectoid Fe–2Mn–C alloys

Carbon enrichment in untransformed austenite at the end of Mn partitionless growth of ferrite for Fe–2Mn–(0.05, 0.14)C (mass%) alloys isothermally transformed in the temperature range 873–998 K was measured using field-emission electron probe microanalysis to reveal its dependence on the transformation temperature, nominal carbon content and prior austenite grain size. The PLE/NPLE model gives much better predictions than the PE model for carbon enrichment in untransformed austenite at the end of partitionless growth. Carbon enrichment could be increased by reducing the prior austenite grain size. Furthermore, carbon enrichment shifted from the PLE/NPLE transition line to the T₀ line on lowering the transformation temperature. This shift is probably attributed to the solute drag effect and/or to the finite interface mobility, both of which vary with the transformation temperature.     

Ultra fine finishing of diamond tools by ion beams

The principles of ion beam machining and its apparatus are described, and several examples are given of ultra fine forming or sharpening diamond tools by ion beams. Problems, such as a sub-micrometre tip radius or elliptical tips are discussed, and it is concluded that ion beam machining combined with mechanical pre-finishing can be useful for forming or sharpening diamond tools.