A Simplified Determination of Microquantity of Carbon in Iron, Steel and Ferro-Alloy

In order to determine rapidly and accurately microquantity (below 0.1 per cent) of carbon in iron, steel and ferro-alloys, 0.5～1g of the sample was taken, carbon dioxide gas generated by its combustion was solidified by passing through a trap chilled in liquid air, and the solid carbon dioxide was vaporized and allowed to permeate a definite vacuum, the volume being read on the manometer. The analytical results by this method agreed well with those by gravimetric method within a mean standard deviation of 0.0005 per cent in low-carbon steel, and well agreeing results were also obtained with chromium metal and manganese metal. The time required for the analysis by this method was 24～26 minutes

Enzymatic Conversion of Cypridina Luciferyl Sulfate to Cypridina Luciferin with Coenzyme A as a Sulfate Acceptor in Cypridina (Vargula) hilgendorfii

In the luminous ostracod Cypridina (presently Vargula) hilgendorfii, Cypridina luciferyl sulfate (3-enol sulfate of Cypridina luciferin) is converted to Cypridina luciferin by a sulfotransferase with 3′-phosphoadenosine-5′-phosphate (PAP) as a sulfate acceptor. The resultant Cypridina luciferin is used for the luciferase–luciferin reaction of Cypridina to emit blue light. The luminescence stimulation with major organic cofactors was examined using the crude extracts of Cypridina specimens, and we found that the addition of coenzyme A (CoA) to the crude extracts significantly stimulated luminescence intensity. Further, the light-emitting source in the crude extracts stimulated with CoA was identified as Cypridina luciferyl sulfate, and we demonstrated that CoA could act as a sulfate acceptor from Cypridina luciferyl sulfate. In addition, the sulfate group of Cypridina luciferyl sulfate was also transferred to adenosine 5′-monophosphate (5′-AMP) and adenosine 3′-monophosphate (3′-AMP) by a sulfotransferase. The sulfated products corresponding to CoA, 5′-AMP, and 3′-AMP were identified using mass spectrometry. This is the first report that CoA can act as a sulfate acceptor in a sulfotransferase reaction

Noninvasive and safe cell viability assay for Euglena gracilis using natural food pigment

Noninvasive and safe cell viability assay is required in many fields such as regenerative medicine, genetic engineering, single-cell analysis, and microbial food culture. In this case, a safe and inexpensive method which is a small load on cells and the environment is preferable without requiring expensive and space-consuming equipment and a technician to operate. We examined eight typical natural food pigments to find Monascus pigment (MP) or anthocyanin pigment (AP) works as a good viability indicator of dye exclusion test (DET) for Euglena gracilis which is an edible photosynthetic green microalga. This is the first report using natural food pigments as cell viability assay. Euglena gracilis stained by MP or AP can be visually judged with a bright field microscope. This was spectrally confirmed by scan-free, non-invasive absorbance spectral imaging A(x, y, λ) microscopy of single live cells and principal component analysis (PCA). To confirm the ability of staining dead cells and examine the load on the cells, these two natural pigments were compared with trypan blue (TB) and methylene blue (MP), which are synthetic dyes conventionally used for DET. As a result, MP and AP had as good ability of staining dead cells treated with microwave as TB and MB and showed faster and more uniform staining for dead cells in benzalkonium chloride than them. The growth curve and the ratio of dead cells in the culture showed that the synthetic dyes inhibit the growth of E. gracilis, but the natural pigments do not. As the cell density increased, however, AP increased the ratio of stained cells, which was prevented by the addition of glucose. MP can stain dead cells in a shorter time than AP, while AP is more stable in color against long-term irradiation of intense light than MP. Due to the low toxicity of these pigments, viability of cells in culture can be monitored with them over a long period

Mechanical loss of a multilayer tantala/silica coating on a sapphire disk at cryogenic temperatures: toward the KAGRA gravitational wave detector

We report the results of a new experimental setup to measure the mechanical loss of coating layers on a thin sapphire disk at cryogenic temperatures. Some of the authors previously reported that there was no temperature dependence of the mechanical loss from a multilayer tantala/silica coating on a sapphire disk, both before and after heat treatment, although some reports indicate that Ta<sub>2</sub>O<sub>5</sub> and SiO<sub>2</sub> layers annealed at 600 °C have loss peaks near 20 K. Since KAGRA—the Japanese gravitational-wave detector, currently under construction—will be operated at 20 K and have coated sapphire mirrors, it is very important to clarify the mechanical loss behavior of tantala/silica coatings around this temperature. We carefully investigate a tantala/silica-coated sapphire disk with the new setup, anneal the disk, and then investigate the annealed disk. We find that there is no distinct loss peak both before and after annealing under particular conditions. The mechanical loss for the unannealed disk at 20 K is about 5×10<sup>−4</sup>, as previously reported, while that for the annealed disk is approximately 6.4×10<sup>−4</sup>

Cross-conjugated isothianaphthene quinoids: a versatile strategy for controlling electronic structures

The elucidation of new structure–property relationships in π-conjugated molecules bearing quinoidal moieties is of relevance because of their use in organic electronics applications and their traditional assimilation as models of doped conducting polymers. Quinoidal oligothiophenes are ground state electronic hybrids between closed-shell Kekulé quinoidal and open-shell aromatic diradicaloid forms. The prominent contribution of the diradical character in longer oligomers beyond thiophene 4-mers results in a low stability, thereby limiting the ability to tune their properties. Thus, the control of these quinoidal/aromatic contributions is an important prerequisite to develop long quinoidal oligothiophenes. To address this problem, a series of quinoidal pentathiophenes with benzene-annelated isothianaphthene units were designed and successfully synthesized as stable structures. Combined molecular spectroscopies and theoretical modelling indicated that cross-conjugation appears upon the introduction of multiple benzene-annelated units, and that the number and position of the benzene-annelated units have a significant influence on the quinoidal/aromatic/cross-conjugated electronic structures. The newly developed quinoidal pentathiophenes functioned as organic semiconducting materials in transistor and near infrared phototransistor devices. This study demonstrates that modification of the cross-conjugated quinoidal structure is a promising strategy for fine-tuning electronic structures in π-extended quinoidal systems, which could help us to understand unique π-electronic features and to develop novel organic electronic materials.This work was supported by JSPS KAKENHI (20H02814, 20K21224, 20H05841, 20KK0123, 19K15505, 20H04639, 20K15352, 21K05213, 20H00379, 20H05833, and 20K15261), CREST (J205101030), NEDO (21500248-0), and “Dynamic Alliance for Open Innovation Bridging Human, Environmental and Materials” from The Ministry of Education, Culture, Sports, Science and Technology, Japan. We are grateful to Prof. Toshihiro Ohnishi for helpful discussion. The authors thank the Spanish Ministry of Science, Innovation and Universities MCIU and MINECO/FEDER of the Spanish Government (project PGC2018-098533-B-100), the Ministry of Science and Technology of the Spanish Government (project RED2018-102626-T) and the Junta de Andalucía, Spain (UMA18FEDERJA057). We also thank the Research Central Services (SCAI) of the University of Málaga. // Funding for open access charge: Universidad de Málag

AJK2011-36009

ABSTRACT We propose a novel cell stimulation device for the analysis of cell responses to chemical stimuli. In order to deliver chemical substances to target single cells, we developed a microfluidic device having microchannels and apertures in the side wall to subject stimuli to laterally trapped cells. The channels were designed to allow simple flow control with single syringe pump. We demonstrated single cell trapping and culturing of pancreatic cell with the device. To test its feasibility in cell stimulation assay, intracellular response of the cell to glucose stimulation was demonstrated

A guiding role of the Arabidopsis circadian clock in cell differentiation revealed by time-series single-cell RNA sequencing

Circadian rhythms and progression of cell differentiation are closely coupled in multicellular organisms. However, whether establishment of circadian rhythms regulates cell differentiation or vice versa has not been elucidated due to technical limitations. Here, we exploit high cell fate plasticity of plant cells to perform single-cell RNA sequencing during the entire process of cell differentiation. By analyzing reconstructed actual time series of the differentiation processes at single-cell resolution using a method we developed (PeakMatch), we find that the expression profile of clock genes is changed prior to cell differentiation, including induction of the clock gene LUX ARRYTHMO (LUX). ChIP sequencing analysis reveals that LUX induction in early differentiating cells directly targets genes involved in cell-cycle progression to regulate cell differentiation. Taken together, these results not only reveal a guiding role of the plant circadian clock in cell differentiation but also provide an approach for time-series analysis at single-cell resolution

Status of 48Ca double beta decay search in CANDLES

We study a strategy to reduce veto-time in the search for neutrino-less double-beta decay (0υββ) with CANDLES-III system. We develop a new likelihood analysis and apply it to our new Run010 data. We show that we can increase the un-vetoed live-time by 11.8%. Thanks to this improvements, We expect to increase a limit on the life-time of 0υββ by a factor of three by analyzing both Run009 and Run010 data