Simultaneous induction of calcium transients in embryoid bodies using microfabricated electrode substrates

Precise control of differentiation processes of pluripotent stem cells is a key component for the further development of regenerative medicine. For this purpose, combining a cell-aggregate-size treatment for regulating intercellular signal transmissions and an electrical stimulation technique for inducing cellular responses is a promising approach. In the present study, we developed microfabricated electrode substrates that allow simultaneous stimulation of embryoid bodies (EBs) of P19 cells. Mouse embryonal carcinoma P19 cells can be induced to differentiate into three germ layers and serve as a promising stem cell model. Microcavity–array patterns were fabricated onto indium–tin–oxide (ITO) substrates using a standard photo-lithography technique, and uniform-sized EBs of P19 cells were inserted into each microcavity. Electrical stimulation was applied to the EBs through substrate electrodes and stimulus-induced intracellular calcium transients were monitored. We confirmed that the developed electrode device could simultaneously stimulate smaller (200 μm diameter) and larger (500 μm diameter) EBs inserted in the microcavities and induce specific spatio-temporal patterns of intracellular calcium transients in the EBs with fine reproducibility. We concluded that the developed microcavity array with embedded electrodes could simultaneously and effectively stimulate uniform-sized EBs inserted in it. Therefore, it is a promising experimental tool for precisely controlling cell differentiation processes.     

Use of multivariate analysis for the improvement in prediction accuracy of bacterial aerobic plate count by flow cytometry

Flow cytometry (FCM) and aerobic plate count (APC) by the culture method were performed on green tea samples spiked with Escherichia coli type strain NCTC9001 (ATCC11775) solutions of different concentrations. In FCM, fluorescence signals from multiple stained bacteria and other fluorophores are detected using detector channels, and recorded as events with a voltage at each channel. FCM data were analyzed in two ways: conventional and multivariate analysis. In the former, the number of events with voltages larger than the defined threshold values was regarded as the predicted APC. In the latter, voltage histograms of all channels were obtained and merged horizontally to serve as explanatory variables. Then a partial least squares regression (PLSR) model was built to predict APC from the histogram data. The coefficient of determination (R²) and the root mean square error (RMSE) between APC by the culture method and that predicted by conventional FCM were 0.916 and 1.08 cfu/ml². The APC values predicted by the PLSR model and those measured were in good agreement with R² of 0.982 and RMSE of 0.417 cfu/ml, which verified the potential of the proposed method for improving APC prediction accuracy by FCM.     

Tailoring Multistimuli Responsive Micropatterns Activated by Various Mechanical Modes

A versatile wrinkle-based micropatterned system with a film-substrate structure is proposed, which contains a hydrophilic film of polyvinyl alcohol or its composite with laponite and a hydrophobic substrate of polydimethylsiloxane or its composite with carbon black. The wrinkled system features high design flexibility and multistimuli responsiveness, which can be activated by various mechanical methods, including vertical press or scratch, gentle stretch-and-release, bend, or analogous magneto-mechanical and electro-mechanical modes. The resultant wrinkles possess 1) instantaneous and reversible strain/moisture/light responsive optical modulation; 2) tunable dynamics for the aforementioned strain/moisture/light response;3) tailorable amplitude/wavelength; 4) unique surface morphologies from the coupling of wrinkles and cracks; 5) excellent reversibility and durability. A variety of applications are demonstrated based on this system, including 1) a moisture erasable highly sensitive pressure responsive device and pattern replicator with a high fidelity; 2) a moisture erasable scratch/magneto-mechanical re-writable tablet; 3) an electro-mechanical controllable smart window with an ultra-sensitive strain responsive transmittance modulation and a low operating voltage; 4) various types of strain responsive, moisture erasable, and laser writable information recording/encryption devices. This work provides new routes for designing innovative wrinkled systems triggered by diverse mechanical fashions and can decode multiple environmental stimuli into optical signals for widespread application.     

Preferable structure of poly(ethylene glycol) for grafting onto a cellulosic membrane to increase hemocompatibility without reduction in solute permeability of the membrane

Swelling layers formed by poly(ethylene glycol) (PEG) chains grafted onto surfaces of a cellulosic membrane are known to improve hemocompatibility of the membrane. Three types of hemodialysis membranes were derived from the same regenerated-cellulose hollow-fiber membrane by grafting PEG with different formulas onto the surfaces to clarify the influence of the grafted PEG chains on solute permeability of the membranes. Determination of volume fractions of nonfreezing water contained in the membranes by differential scanning calorimetry revealed that most of the PEG chains were grafted onto the external surfaces and less into the pores in the membranes. Permeability of vitamin B₁₂ for the PEG-grafted membranes except for the one with the shortest PEG chains was reduced as compared with the original membrane, while that of tritium-labeled water for all the PEG-grafted membranes was the same as that of the original membrane. Structural parameters only of the PEG-grafted membrane with the largest alkyl groups at the terminal of the PEG chains were different from those of the other PEG-grafted and original membranes. The shorter PEG chains with the larger terminal alkyl groups are suitable for grafting onto a cellulosic membrane to increase hemocompatibility of the membrane without significant reduction in the solute permeability of the membrane. © 1995 John Wiley & Sons, Inc.     

Reaction of layered carbon fluorides CxF (x = 2.5-3.6) and hydrogen

The layered carbon fluorides C_xF (x = 2.5, 2.8, 3.6), generally classified as fluorine-graphite intercalation compounds, were heat-treated in hydrogen gas. These fluorides are more reactive with hydrogen compared to (CF)_n and (C₂F)_n. Reduction of C_xF to graphite-like carbon starts at about 573 K, and proceeds gradually along with the elevation of temperature. Fluorine atoms in C_xF are eliminated as HF in the reduction process without being substituted by hydrogen atoms. Systematic difference was not found in the average crystallite sizes of the carbon material prepared from C_xF by the reduction with hydrogen and that by the pyrolysis in vacuum. On the other hand, interlayer distance and fluorine content of the former are smaller than those of the latter. In the case that the C_xF precursor maintains a large particle size, the reduced carbon as well as the pyrolytically prepared carbon possesses a foam-like shape due to the exfoliation during the heat treatment.     

Rupture of the aortic arch due to bacterial aortitis--a case report of a patient undergoing successful surgical therapy

A 59-year-old man came to our hospital because of hoarseness and chest pain. On initial examination, a chest roentgenogram showed no abnormality. Three weeks later, while the patient was waiting to undergo a computed tomographic (CT) scan, shock suddenly developed. A chest roentgenogram revealed an extremely wide mediastinal shadow. A CT scan revealed rupture of the aortic arch near the base of the left subclavian artery, although there was no aneurysm. An emergency operation was performed. The ruptured aortic arch was replaced with an artificial graft during cardiopulmonary bypass with selective cerebral perfusion. A diagnosis of infectious aortitis was not made until a pathologist discovered colonies of gram-positive cocci in the resected specimen of the aortic arch. Postoperatively, antibiotics were administered for only four days. The patient is well about two years after the operation. Infectious aortitis had a very low prevalence and rarely involves the thoracic aorta. However, we should be aware of that it may cause aortic rupture even in the absence of aneurysmal dilatation.     

Wortmannin, a specific inhibitor of phosphatidylinositol-3-kinase, inhibits erythropoietin-induced erythroid differentiation of K562 cells

We report a large kindred in which a punctate palmoplantar keratoderma (PPK) is associated with malignancy, including Hodgkin's disease, renal, breast, pancreatic and colonic adenocarcinomas. The family was traced through four generations, and over 320 individuals were identified, of whom 49 had punctate PPK. The punctate PPK appeared to be inherited as an autosomal dominant trait with variable penetrance. Ten of the 43 adults (23%) with punctate PPK developed malignancies, and five of these developed before the age of 50. Of the 271 unaffected individuals, six (2%) have developed malignancies, one prior to the age of 50. The association of keratoderma and malignancy is discussed.     

Temperature dependence of luminescence from a silica glass under in-reactor irradiation

We have investigated in-reactor luminescence (IRL) from a silica glass at temperatures ranging from 100 K to 250 K. The IRL consists mainly of a broad emission band peaked at 2.7 eV assigned to oxygen deficient centers produced in the silica glass under the in-reactor irradiation. The 2.7 eV emission intensity linearly increased with the irradiation time and its increasing rate was larger for higher irradiation temperatures. However, this temperature dependence is inconsistent with that for the defect production rate and the cause is not clear at present. The initial intensity of the 2.7 eV IRL band increases with temperature, showing an activation energy of ca 21 meV. This value is much lower than those observed in the temperature dependence of the 2.7 eV photoluminescence (PL) and the cathodeluminescence (CL) induced by 8 keV electron irradiation. These results suggest that in IRL, some electrons excited to higher energy levels than the luminescence level are likely transferred to the luminescence state without thermal activation, resulting in a lower activation energy in their temperature dependence.     

Temporal changes in vertical distribution of 137Cs in litter and soils in mixed deciduous forests in Fukushima,Japan

Downward migration of ¹³⁷Cs in soils was studied in three mixed deciduous forests c.a. 40 km northwest of the Fukushima Daiichi Nuclear power plant (FDNPP), Japan. We selected three different types of forest regarding to environmental condition such as slope inclinations and snow coverage conditions at the time of deposition. We examined temporal changes in the vertical distribution of ¹³⁷Cs from litter layers to 10 cm soil depth for two years (2.3 to 4.3 years after the FDNPP accident in 2011). At all three study sites, the ¹³⁷Cs in the litter layer had largely migrated to surface soil by 2013. After 2014, about 80% of the ¹³⁷Cs in forest soils (litter layer to 10 cm soil depth) remained within 0–5 cm soil layer. The vertical distribution had not changed substantially since 2014, suggesting that changes to the downward migration rates of ¹³⁷Cs in soils drastically decreased with time. In addition, small amounts of migrating ¹³⁷Cs could not be detected by the present method because there was a large spatial variation in the distribution of soil ¹³⁷Cs. The results showed similar patterns of soil ¹³⁷Cs distribution among the three study sites although there were differences in the environmental conditions.