Changing Pattern of Deiminated Proteins in Developing Human Epidermis

Peptidylarginine deiminases are widely distributed, calcium-ion-dependent enzymes that convert arginine residues of proteins into citrulline residues. This reaction, deimination, is thought to be an important event during the final stage of epidermal differentiation, possibly associated with integration and disintegration of keratin filaments. To elucidate the possible roles of protein deimination during human epidermal development we investigated localization of deiminated proteins using anti-citrulline peptide antibody, which preferentially recognizes citrulline residues in the V subdomains of keratin 1, and anti-chemically modified citrulline antibody, which enables detection of citrulline residues independent of amino acid sequences. Anti-chemically modified citrulline antibody, but not anti-citrulline peptide antibody stained the periderm in two-layered epidermis of 49 d and 57 d estimated gestational age. In the stratified epidermis of 88 d, 96 d, and 108 d estimated gestational age fetal skin, anti-citrulline peptide antibody and anti-chemically modified citrulline antibody staining was seen in the periderm and intermediate cell layers. After periderm cells regressed and keratinization began in the interfollicular epidermis, anti-citrulline peptide antibody and anti-chemically modified citrulline antibody were restricted to the cornified cell layers of the interfollicular epidermis, similar to the distribution patterns of that in adult epidermis. Postembedding immunoelectron microscopy showed anti-citrulline peptide antibody immunogold labeling over the cytoplasmic intermediate filament network in the periderm and the intermediate cell layers. These results demonstrate an orderly formation of deiminated proteins in different layers of embryonic epidermis and suggest important roles for peptidylarginine deiminases in human epidermal morphogenesis

Coarse grained DEM simulation of non-spherical and poly-dispersed particles using Scaled-Up Particle (SUP) model

In this work, the validity of the Scaled-Up Particle (SUP) model, which is a novel coarse grain model for Discrete Element Method (DEM), is examined to simulate a flow of non-spherical and poly-dispersed particles. The SUP model is based on the authors’ previous work and the scaling law is derived from the continuum assumption of an arbitrary particles flow. We discuss that the model is applicable not only to spherical and mono-dispersed particles, as is the case tested in the previous work, but also to non-spherical and poly-dispersed particles. Simulations of various systems are performed such as compression of a particle bed, heap formation, high shear mixer, large scale rotary drum and V-mixer. It is shown that the results obtained from the SUP model are in both qualitative and quantitative agreement with those from the original particles as long as the resolution is sufficiently high.Washino K., Chan E.L., Nishida Y., et al. Coarse grained DEM simulation of non-spherical and poly-dispersed particles using Scaled-Up Particle (SUP) model. Powder Technology 426, 118676 (2023); https://doi.org/10.1016/j.powtec.2023.118676

Magnetic field generation by pointwise zero-helicity three-dimensional steady flow of incompressible electrically conducting fluid

We introduce six families of three-dimensional space-periodic steady solenoidal flows, whose kinetic helicity density is zero at any point. Four families are analytically defined. Flows in four families have zero helicity spectrum. Sample flows from five families are used to demonstrate numerically that neither zero kinetic helicity density, nor zero helicity spectrum prohibit generation of large-scale magnetic field by the two most prominent dynamo mechanisms: the magnetic $\alpha$-effect and negative eddy diffusivity. Our computations also attest that such flows often generate small-scale field for sufficiently small magnetic molecular diffusivity. These findings indicate that kinetic helicity and helicity spectrum are not the quantities controlling the dynamo properties of a flow regardless of whether scale separation is present or not.Comment: 37 pages, 11 figures, 54 reference

Guest‐selective gate‐opening by pore engineering of two‐dimensional Kagomè lattice porous coordination polymers

Porous coordination polymers (PCPs) with pore decoration have been used as materials for excellent storage and separation functions. The cooperative properties of flexible PCPs can be utilized to achieve the separation of mixtures of gaseous molecules having highly similar properties. The key to efficient molecular recognition and separation lies in increasing the degrees of freedom of the structure without sacrificing the stability of the system. However, the mechanism study of such behavior is still scarce in the literature. Here, we focused on PCPs with two-dimensional Kagomè lattice structures and functionalized the pores with various alkoxy pendant groups; this facilitated systematic tuning of the pore aperture size, the interlayer spacing, as well as the interactions between the host and adsorbed molecules. The combination of characterization techniques allowed us to observe a unique deformation of the lattice upon gas sorption, allowing the separation of gas molecules with similar physicochemical properties, such as propane and propylene

Downregulation of barley ferulate 5-hydroxylase dramatically alters straw lignin structure without impact on mechanical properties

Barley is a major cereal crop for temperate climates, and a diploid genetic model for polyploid wheat. Cereal straw biomass is an attractive source of feedstock for green technologies but lignin, a key determinant of feedstock recalcitrance, complicates bio-conversion processes. However, manipulating lignin content to improve the conversion process could negatively affect agronomic traits. An alternative approach is to manipulate lignin composition which influences the physical and chemical properties of straw. This study validates the function of a barley ferulate 5-hydroxylase gene and demonstrates that its downregulation using the RNA-interference approach substantially impacts lignin composition. We identified five barley genes having putative ferulate 5-hydroxylase activity. Downregulation of HvF5H1 substantially reduced the lignin syringyl/guaiacyl (S/G) ratio in straw while the lignin content, straw mechanical properties, plant growth habit, and grain characteristics all remained unaffected. Metabolic profiling revealed significant changes in the abundance of 173 features in the HvF5H1-RNAi lines. The drastic changes in the lignin polymer of transgenic lines highlight the plasticity of barley lignification processes and the associated potential for manipulating and improving lignocellulosic biomass as a feedstock for green technologies. On the other hand, our results highlight some differences between the lignin biosynthetic pathway in barley, a temperate climate grass, and the warm climate grass, rice, and underscore potential diversity in the lignin biosynthetic pathways in grasses

DHPLC Denaturing High Performance Liquid Chromatography オ モチイタ ダンジョ シキベツ

A novel method for sex identification using a denaturing high performance liquid chromatography (DHPLC) system is described. Among many methods for identifying sex, the most popular and credible system has been the polymerase chain reaction (PCR) method using the nucleotide primer sets of the amelogenin gene, which is shared on both the X and Y chromosomes. In this conventional method, the judgment depends on the detection of the size difference between the PCR products derived from the X and Y chromosomes. In this study, we adopted DHPLC to detect the difference by checking heteroduplex formation between the products, which enabled us to shorten the PCR products down to 45 bp and the separation time within 8 minutes. This new system may have a wide application in many different fields such as forensic medicine, prenatal diagnosis, inbreeding of animals and anthropology

Downregulation of barley ferulate 5-hydroxylase dramatically alters straw lignin structure without impact on mechanical properties

Barley is a major cereal crop for temperate climates, and a diploid genetic model for polyploid wheat. Cereal straw biomass is an attractive source of feedstock for green technologies but lignin, a key determinant of feedstock recalcitrance, complicates bio-conversion processes. However, manipulating lignin content to improve the conversion process could negatively affect agronomic traits. An alternative approach is to manipulate lignin composition which influences the physical and chemical properties of straw. This study validates the function of a barley ferulate 5-hydroxylase gene and demonstrates that its downregulation using the RNA-interference approach substantially impacts lignin composition. We identified five barley genes having putative ferulate 5-hydroxylase activity. Downregulation of HvF5H1 substantially reduced the lignin syringyl/guaiacyl (S/G) ratio in straw while the lignin content, straw mechanical properties, plant growth habit, and grain characteristics all remained unaffected. Metabolic profiling revealed significant changes in the abundance of 173 features in the HvF5H1-RNAi lines. The drastic changes in the lignin polymer of transgenic lines highlight the plasticity of barley lignification processes and the associated potential for manipulating and improving lignocellulosic biomass as a feedstock for green technologies. On the other hand, our results highlight some differences between the lignin biosynthetic pathway in barley, a temperate climate grass, and the warm climate grass, rice, and underscore potential diversity in the lignin biosynthetic pathways in grasses

RNAi-suppression of barley caffeic acid O-methyltransferase modifies lignin despite redundancy in the gene family

Caffeic acid O-methyltransferase (COMT), the lignin biosynthesis gene modified in many brown-midrib high-digestibility mutants of maize and sorghum, was targeted for downregulation in the small grain temperate cereal, barley (Hordeum vulgare), to improve straw properties. Phylogenetic and expression analyses identified the barley COMT orthologue(s) expressed in stems, defining a larger gene family than in brachypodium or rice with three COMT genes expressed in lignifying tissues. RNAi significantly reduced stem COMT protein and enzyme activity, and modestly reduced stem lignin content while dramatically changing lignin structure. Lignin syringyl-to-guaiacyl ratio was reduced by ~50%, the 5-hydroxyguaiacyl (5-OH-G) unit incorporated into lignin at 10--15-fold higher levels than normal, and the amount of p-coumaric acid ester-linked to cell walls was reduced by ~50%. No brown-midrib phenotype was observed in any RNAi line despite significant COMT suppression and altered lignin. The novel COMT gene family structure in barley highlights the dynamic nature of grass genomes. Redundancy in barley COMTs may explain the absence of brown-midrib mutants in barley and wheat. The barley COMT RNAi lines nevertheless have the potential to be exploited for bioenergy applications and as animal feed

Direct Observation of the Extended Molecular Atmosphere of o Cet by Differential Spectral Imaging with an Adaptive Optics System

We present new measurements of the diameter of o Cet (Mira) as a function of wavelength in the 2.2 micron atmospheric window using the adaptive optics system and the infrared camera and spectrograph mounted on the Subaru Telescope. We found that the angular size of the star at the wavelengths of CO and H2O absorption lines were up to twice as large as the continuum photosphere. This size difference is attributable to the optically thick CO and H2O molecular layers surrounding the photosphere. This measurement is the first direct differential spectroscopic imaging of stellar extension that resolves individual molecular lines with high spectral-resolution observations. This observation technique is extremely sensitive to differences in spatial profiles at different wavelengths; we show that a difference in diameter much smaller than the point spread function can be measured.Comment: 6 pages, 3 figures, accepted for publication in PAS

Tissue and cell-specific transcriptomes in cotton reveal the subtleties of gene regulation underlying the diversity of plant secondary cell walls

Background Knowledge of plant secondary cell wall (SCW) regulation and deposition is mainly based on the Arabidopsis model of a ‘typical’ lignocellulosic SCW. However, SCWs in other plants can vary from this. The SCW of mature cotton seed fibres is highly cellulosic and lacks lignification whereas xylem SCWs are lignocellulosic. We used cotton as a model to study different SCWs and the expression of the genes involved in their formation via RNA deep sequencing and chemical analysis of stem and seed fibre. Results Transcriptome comparisons from cotton xylem and pith as well as from a developmental series of seed fibres revealed tissue-specific and developmentally regulated expression of several NAC transcription factors some of which are likely to be important as top tier regulators of SCW formation in xylem and/or seed fibre. A so far undescribed hierarchy was identified between the top tier NAC transcription factors SND1-like and NST1/2 in cotton. Key SCW MYB transcription factors, homologs of Arabidopsis MYB46/83, were practically absent in cotton stem xylem. Lack of expression of other lignin-specific MYBs in seed fibre relative to xylem could account for the lack of lignin deposition in seed fibre. Expression of a MYB103 homolog correlated with temporal expression of SCW CesAs and cellulose synthesis in seed fibres. FLAs were highly expressed and may be important structural components of seed fibre SCWs. Finally, we made the unexpected observation that cell walls in the pith of cotton stems contained lignin and had a higher S:G ratio than in xylem, despite that tissue’s lacking many of the gene transcripts normally associated with lignin biosynthesis. Conclusions Our study in cotton confirmed some features of the currently accepted gene regulatory cascade for ‘typical’ plant SCWs, but also revealed substantial differences, especially with key downstream NACs and MYBs. The lignocellulosic SCW of cotton xylem appears to be achieved differently from that in Arabidopsis. Pith cell walls in cotton stems are compositionally very different from that reported for other plant species, including Arabidopsis. The current definition of a ‘typical’ primary or secondary cell wall might not be applicable to all cell types in all plant species.CPM was funded by Cotton Breeding Australia, a joint venture between Cotton Seed Distributors and CSIRO (Project No. CBA19). HB was funded by the CSIRO’s Office of the Chief Executive (OCE) Postdoctoral Fellowship program. YT and JR were funded in part by Stanford University’s Global Climate and Energy Program, and in part by the DOE Great Lakes Bioenergy Research Center (DOE BER Office of Science, DE-FC02–07ER6449