An Engineered Biliverdin-Compatible Cyanobacteriochrome Enables a Unique Ultrafast Reversible Photoswitching Pathway

Cyanobacteriochromes (CBCRs) are promising optogenetic tools for their diverse absorption properties with a single compact cofactor-binding domain. We previously uncovered the ultrafast reversible photoswitching dynamics of a red/green photoreceptor AnPixJg2, which binds phycocyanobilin (PCB) that is unavailable in mammalian cells. Biliverdin (BV) is a mammalian cofactor with a similar structure to PCB but exhibits redder absorption. To improve the AnPixJg2 feasibility in mammalian applications, AnPixJg2_BV4 with only four mutations has been engineered to incorporate BV. Herein, we implemented femtosecond transient absorption (fs-TA) and ground state femtosecond stimulated Raman spectroscopy (GS-FSRS) to uncover transient electronic dynamics on molecular time scales and key structural motions responsible for the photoconversion of AnPixJg2_BV4 with PCB (Bpcb) and BV (Bbv) cofactors in comparison with the parent AnPixJg2 (Apcb). Bpcb adopts the same photoconversion scheme as Apcb, while BV4 mutations create a less bulky environment around the cofactor D ring that promotes a faster twist. The engineered Bbv employs a reversible clockwise/counterclockwise photoswitching that requires a two-step twist on ~5 and 35 picosecond (ps) time scales. The primary forward P_fr → P_o transition displays equal amplitude weights between the two processes before reaching a conical intersection. In contrast, the primary reverse P_o → P_fr transition shows a 2:1 weight ratio of the ~35 ps over 5 ps component, implying notable changes to the D-ring-twisting pathway. Moreover, we performed pre-resonance GS-FSRS and quantum calculations to identify the Bbv vibrational marker bands at ~659,797, and 1225 cm⁻¹. These modes reveal a stronger H-bonding network around the BV cofactor A ring with BV4 mutations, corroborating the D-ring-dominant reversible photoswitching pathway in the excited state. Implementation of BV4 mutations in other PCB-binding GAF domains like AnPixJg4, AM1_1870g3, and NpF2164g5 could promote similar efficient reversible photoswitching for more directional bioimaging and optogenetic applications, and inspire other bioengineering advances.     

Investigation of laser consolidation process for metal powder by two-color pyrometer and high-speed video camera

This paper deals with the measurement of surface temperature on metal powder during the laser consolidation process with two-color pyrometer. Additionally, the aspect of selective laser sintering (SLS) and selective laser melting (SLM) of metal powder is visualized with high speed video camera. As a result, the surface temperature during the laser irradiation was ranged 1520–1810 °C and the consolidation phenomena was classified according to the melting point of metal powder. The metal powder at the heating process cohered intermittently to the melt pool although the laser beam was continuously irradiated to the powder surface.     

Excess estrogen sulfoconjugation as the possible cause for a poor sign of parturition in pregnant cows carrying somatic cell clone fetuses

We conducted this study to elucidate a factor causing a poor sign of parturition and prolonged gestation, which is frequently observed in cows carrying somatic clone fetuses. Pre-partum rises in concentrations of plasma estrone and estradiol-17beta in the recipient cows pregnant with clones were subtle. By contrast, the plasma concentration of estrone sulfate in clone pregnancies increased gradually from pre-initiation of parturition induction whereas control cows that received in vivo-derived embryos showed a significant increase at parturition. Therefore, in clone pregnancies, the ratio of estrone/estrone sulfate was low during the pre-partum period compared with control. Messenger RNA expression of estrogen sulfotransferase (SULT1E1) in the placenta at parturition was significantly higher in clone pregnancies than control pregnancies and was localized in binucleate cells (BNC). SULT1E1 mRNA abundance was negatively and positively correlated with concentrations of maternal estrone and estrone sulfate at parturition respectively. Messenger RNA expressions of estrogen sulfatase (STS) and aromatase (CYP19) were similar between clone and control pregnancies and were localized in BNC and caruncular epithelial cells. STS and CYP19 mRNA abundances showed positive correlations with maternal estradiol-17beta concentration. The population of BNC in the placenta did not differ between clone and control pregnancies. Plasma cortisol concentration of vaginally delivered newborn clone calves was comparable with those of control, although cesarean section delivered clone calves showed a low concentration. These results suggest that excess estrogen sulfoconjugation is the reason for the perturbed low ratio of active to inactive estrogens and the resulting hormonal imbalance contributes to the lack of overt signs of readiness for parturition in cows pregnant with clones.     

A study of the volume and bounce decrease in hair with ageing using bending elasticity measurements

Hair diameters and bending stresses were measured for chemically untreated regions of hair close to the scalp, collected from 38 Japanese female (age range: 26–51) and 35 Caucasian female (age range: 20–40) volunteers. For the bending stress measurements, a new methodology to obtain averaged Young's modulus has been developed in which 50 fibres are measured together to give reproducible data, overcoming the individual differences in each hair fibre. By comparing the resulting data of the fibres collected from the panelists younger than 40 years of age, no significant difference in Young's modulus between Japanese and Caucasian hair was recognized, and the modulus was c. 10 GPa, regardless of the hair diameter. On the other hand, there existed cases in which fibres showed lower Young's moduli, and these fibres were from senior panelists who were conscious of the decrease in hair volume and bounce with ageing. This result indicates that the decrease in volume and bounce in hair with ageing is not only caused by a decrease in hair diameter and fewer hairs per square centimetre of scalp but also by some internal change in the hair.     

Physicochemical properties and photocatalytic hydrogen evolution of TiO2 films prepared by sol–gel processes

Anatase TiO₂ films were obtained on glass substrates using a sol–gel method using titanium isopropoxide as a precursor. The thickness of the film was about 140 nm for one coating, and the thickness is controlled by the number of coating cycles. The spectra of UV-VIS absorption indicated that the absorption edge of the TiO₂ films is ca. 385 nm, corresponding to the band gap energy of 3.20 eV. We obtained TiO₂ films having a high activity for the hydrogen evolution from photocatalytic water cleavage. By loading with 0.3 wt% Pt rate of hydrogen production increases. No influence of film thickness and calcination temperature on the photocatalytic property is observed.     

E-cadherin gene-engineered feeder systems for supporting undifferentiated growth of mouse embryonic stem cells

Conventionally, embryonic stem (ES) cells are cultured on a cell layer of mouse embryonic fibroblasts (MEFs) as feeder cells to support undifferentiated growth of ES cells. In this study, cell–cell interactions between mouse ES and feeder cells were artificially engineered via an epithelial cell adhesion molecule, E-cadherin, whose expression is considerable in ES cells. Mouse mesenchymal STO and NIH3T3 cells that were genetically engineered to express E-cadherin were used in ES cell cultures as feeder cells. ES cells cultured on the E-cadherin-expressing feeder cells maintained the expression of stem cell markers, alkaline phosphatase (AP), Oct3/4, Nanog and Sox2, and the efficiency of AP-positive colony formation was comparable to MEFs, and much better than parental STO and NIH3T3 cells. Furthermore, ES cells maintained on the E-cadherin-expressing feeder cells possessed the ability to differentiate into the three germ layers both in vitro and in vivo. The results indicated that E-cadherin expression in feeder cells could improve the performance of feeder cells, which may be further applicable to create new artificial feeder cell lines.     

Estimation of collection efficiency depended on feed particle concentration for axial flow cyclone dust collector

INTRODUCTIONCyclonedustcollectorisappliedinmanyindustries.Especiallytheaxialflowcycloneisthemostsimpleconstructionanditkeepsthehighreliabilityforthecyclonemaintenance.Thereforetheaxialflowcycloneisveryeasytoarrangewithvariouspipelinesofthepneumatictr...     

Thermal Feasibility Analyses for the 1356MWe High Conversion-Type Innovative Water Reactor for Flexible Fuel Cycle

A new reactor concept of innovative water reactor for flexible fuel cycle (FLWR) is under development at Japan Atomic Energy Agency in cooperation with Japanese reactor suppliers. A design of 1,356 MWe high conversion boiling water reactor-type FLWR core, which has an instantaneous conversion ratio of 1.04, negative void coefficient, high burnup of 65 GWd/t, and 15-month operational cycle length, has been constructed. So far, studies on thermal-hydraulic characteristics have been performed for tight lattice core. Evaluation methods for the critical power and the pressure drop under both the steady and the transient states have been established, and a modified TRAC-BF1 code has been developed for the thermal-hydraulic design of the FLWR. In this paper, the thermal feasibility of the designed 1356MWe FLWR core is analyzed by using the modified TRAC-BF1 code. The analysis is first carried out for the current core design. It is confirmed that no boiling transition (BT) occurs under the steady state. However, the minimum critical power ratio (MCPR) is only about 1.08, and the BT is confirmed occurring under the postulated abnormal transient processes. Therefore, concretizations of the conditions that ensure the thermal feasibility of a natural circulation-type FLWR and a forced circulation-type FLWR are performed. As for the results, for a forced circulation-type FLWR, the operation-limited MCPR (OLMCPR) is 1.32, and the necessary minimum core coolant flow rate is 640 kg/(m²s). For a natural circulation-type FLWR, the OLMCPR is 1.19, and the necessary minimum core coolant flow rate is 560 kg/(m²s).     

Measurement of Liquid Film Thickness by a Fringe Method

A technique was developed and tested to measure the local thickness of a droplet or a liquid film on a surface of an opaque or thick single transparent plate by an interference fringe pattern that was easily formed by reflecting laser lights. Monochromatic epi-illumination through an objective lens of a conventional microscope was provided by a 5 mW or 300 mW laser and a filter to remove the noise caused by laser speckle. The incremental height difference of the liquid layer between neighboring maxima or minima of fringes was evaluated from the wavelength of the laser light and the refractive index of the liquid. Estimation error of a local inclination angle was discussed using ray tracing under parallel illumination approximation. Droplet profiles evaluated from the interferogram that were obtained by the present fringe method agreed well with those by Laser Focus Displacement Meter. Measurement was made to ensure the usefulness of the present technique. It was made clear that the contact angle of a liquid droplet could be obtained precisely and swiftly, even in small size or small contact angle, and that instantaneous three-dimensional profile of a liquid film on a bubble moving in a microchannel could be measured. The fringe method had sufficient potential to obtain more detailed information about three-dimensional characteristics of liquid film in flows such as the generation, breakdown, and growth of waves and the liquid film on a bubble at the beginning of movement.     

Subcooled Flow Boiling in a Minichannel

It has been considered that dry-out occurs easily in boiling heat transfer for a small channel, a mini- or microchannel, because the channel was easily filled with coalescing vapor bubbles. In the present study, the experiments of subcooled flow boiling of water were performed under atmospheric conditions for a horizontal rectangular channel for which the size is 1 mm height and 1 mm width, with a flat heating surface of 10 mm length and 1 mm width placed on the bottom of the channel. The heating surface has a top of copper heating block and is heated by ceramic heaters. In the high heat flux region of nucleate boiling, about 70–80% of the heating surface was covered with a large coalescing bubble and the boiling reached critical heat flux as observed by high-speed video. In the beginning of transition boiling, coalescing bubbles were collapsed to many fine bubbles and microbubble emission boiling was observed at liquid subcooling higher than 30 K. The maximum heat flux obtained was 8 MW/m² (800 W/cm²) at liquid subcooling of higher than 40 K and a liquid velocity of 0.5 m/s. However, the surface temperature was very much higher than that of a centimeter-scale channel. The high-speed video photographs indicated that microbubble emission boiling occurs in the deep transition boiling region.