Millmeter-wave irradiation regulates mRNA-expression and the ubiquitin-proteasome system in wheat exposed to flooding stress.

The irradiation with millimeter-wave (MMW) of wheat seeds promotes root growth under flooding stress; however, its role is not completely clarified. Nuclear proteomics was performed, to reveal the role of MMW irradiation in enhancing flooding tolerance. The purity of nuclear fractions purified from roots was verified. Histone, which is a protein marker for nuclear-purification efficiency, was enriched; and cytosolic ascorbate peroxidase was reduced in the nuclear fraction. The principal-component analysis of proteome displayed that the irradiation of seeds affected nuclear proteins in roots grown under flooding stress. Proteins detected using proteomic analysis were verified using immunoblot analysis. Histone H3 accumulated under flooding stress; however, it decreased to the control level by irradiation. Whereas the ubiquitin accumulated in roots grown under stress when seeds were irradiated. These results suggest that MMW irradiation improves wheat-root growth under flooding stress through the regulation of mRNA-expression level and the ubiquitin-proteasome system. SIGNIFICANCE: To reveal the role of millimeter-wave irradiation in enhancing flooding tolerance in wheat, nuclear proteomics was performed. The principal-component analysis of proteome displayed that irradiation of seeds affected nuclear proteins in roots grown under flooding stress. Proteins detected using proteomic analysis were verified using immunoblot analysis. Histone H3 accumulated under flooding stress; however, it decreased to the control level with irradiation. Whereas the ubiquitin accumulated in roots grown under stress when seeds were irradiated. These results suggest that millimeter-wave irradiation improves wheat-root growth under flooding stress through the regulation of mRNA-expression level and the ubiquitin-proteasome system.

Membrane Proteomics to Understand Enhancement Effects of Millimeter-Wave Irradiation on Wheat Root under Flooding Stress.

Millimeter-wave irradiation of wheat seeds enhances the growth of roots under flooding stress, but its mechanism is not clearly understood. To understand the role of millimeter-wave irradiation on root-growth enhancement, membrane proteomics was performed. Membrane fractions purified from wheat roots were evaluated for purity. H+-ATPase and calnexin, which are protein markers for membrane-purification efficiency, were enriched in a membrane fraction. A principal-component analysis of the proteomic results indicated that the millimeter-wave irradiation of seeds affects membrane proteins in grown roots. Proteins identified using proteomic analysis were confirmed using immunoblot or polymerase chain reaction analyses. The abundance of cellulose synthetase, which is a plasma-membrane protein, decreased under flooding stress; however, it increased with millimeter-wave irradiation. On the other hand, the abundance of calnexin and V-ATPase, which are proteins in the endoplasmic reticulum and vacuolar, increased under flooding stress; however, it decreased with millimeter-wave irradiation. Furthermore, NADH dehydrogenase, which is found in mitochondria membranes, was upregulated due to flooding stress but downregulated following millimeter-wave irradiation even under flooding stress. The ATP content showed a similar trend toward change in NADH dehydrogenase expression. These results suggest that millimeter-wave irradiation improves the root growth of wheat via the transitions of proteins in the plasma membrane, endoplasmic reticulum, vacuolar, and mitochondria.

Proteomic and Biochemical Approaches Elucidate the Role of Millimeter-Wave Irradiation in Wheat Growth under Flooding Stress.

Flooding impairs wheat growth and considerably affects yield productivity worldwide. On the other hand, irradiation with millimeter waves enhanced the growth of chickpea and soybean under flooding stress. In the current work, millimeter-wave irradiation notably enhanced wheat growth, even under flooding stress. To explore the protective mechanisms of millimeter-wave irradiation on wheat under flooding, quantitative proteomics was performed. According to functional categorization, proteins whose abundances were changed significantly with and without irradiation under flooding stress were correlated to glycolysis, reactive-oxygen species scavenging, cell organization, and hormonal metabolism. Immunoblot analysis confirmed that fructose-bisphosphate aldolase and ß tubulin accumulated in root and leaf under flooding; however, even in such condition, their accumulations were recovered to the control level in irradiated wheat. The abundance of ascorbate peroxidase increased in leaf under flooding and recovered to the control level in irradiated wheat. Because the abundance of auxin-related proteins changed with millimeter-wave irradiation, auxin was applied to wheat under flooding, resulting in the application of auxin improving its growth, even in such condition. These results suggest that millimeter-wave irradiation on wheat seeds improves the recovery of plant growth from flooding via the regulation of glycolysis, reactive-oxygen species scavenging, and cell organization. Additionally, millimeter-wave irradiation could promote tolerance against flooding through the regulation of auxin contents in wheat.

Proteomic and Biological Analyses Reveal the Effect on Growth under Flooding Stress of Chickpea Irradiated with Millimeter Waves.

Chickpea cultivated on marginal lands in arid and semiarid tropics is one of the food legumes, and its growth is reduced by flooding stress. Millimeter-wave irradiation has influences on organisms, and it improves the growth of plants such as soybean. To reveal the dynamic effects of millimeter-wave irradiation on chickpea under flooding, gel- and label-free proteomic analysis was conducted. Millimeter-wave irradiation improved chickpea growth and its tolerance to flooding stress. According to functional categorization, oppositely changed proteins were correlated with photosynthesis, fermentation, and protein degradation. Immunoblot analysis confirmed that RuBisCO activase and large subunits decreased in leaves under flooding; however, they are recovered in irradiated chickpea even if it was in this condition. The activity and accumulation of alcohol dehydrogenase increased in roots under flooding; however, this followed the same pattern. Cell death was significantly increased and decreased by flooding on unirradiated and irradiated chickpeas, respectively. These findings suggest that irradiation with millimeter waves on chickpea seeds improves the recovery of plant growth through regulation of photosynthesis in leaves and fermentation in roots. Furthermore, millimeter-wave irradiation might promote chickpea tolerance under flooding via the regulation of cell death.

Evaluation of high-stability optical beats in laser chaos by plasmonic photomixing.

The stability of optical beats in a chaotically oscillating laser is compared to that of a free-running continuous-wave laser using a highly efficient plasmonic photomixer. Using a chaotically oscillating laser diode, stable optical beats are observed over an operation current range of 60-90 mA. The optical spectra are stable even with frequent mode hopping. In contrast, optical beats in a free-running continuous-wave laser are not stable compared to those of a chaotically oscillating laser, because of intermittent hopping of the laser modes. The high stability of chaotically oscillating lasers makes these lasers promising candidates for optical pump sources in terahertz time-domain spectroscopy systems.

Development of a novel fungicide, pyraziflumid.

Pyraziflumid is a novel succinate dehydrogenase inhibitor (SDHI) fungicide discovered and developed by Nihon Nohyaku Co., Ltd. It exhibits excellent fungicidal activities against a broad range of plant diseases and has a favorable safety profile for the Integrated Pest Management (IPM) program. This compound was found by researching the unique chemical derivatives, 3-(trifluoromethyl)pyrazine-2-carboxamides, and has good biological properties, such as preventive, residual and curative activity, and rain-fastness. Pyraziflumid was registered and launched in Japan in 2018. It was registered in South Korea in 2018 and is now under development in other countries. This paper describes the discovery, synthesis, biological activity, safety profile and mode of action of pyraziflumid.

Proteomic Analysis of Irradiation with Millimeter Waves on Soybean Growth under Flooding Conditions.

Improving soybean growth and tolerance under environmental stress is crucial for sustainable development. Millimeter waves are a radio-frequency band with a wavelength range of 1-10 mm that has dynamic effects on organisms. To investigate the potential effects of millimeter-waves irradiation on soybean seedlings, morphological and proteomic analyses were performed. Millimeter-waves irradiation improved the growth of roots/hypocotyl and the tolerance of soybean to flooding stress. Proteomic analysis indicated that the irradiated soybean seedlings recovered under oxidative stress during growth, whereas proteins related to glycolysis and ascorbate/glutathione metabolism were not affected. Immunoblot analysis confirmed the promotive effect of millimeter waves to glycolysis- and redox-related pathways under flooding conditions. Sugar metabolism was suppressed under flooding in unirradiated soybean seedlings, whereas it was activated in the irradiated ones, especially trehalose synthesis. These results suggest that millimeter-waves irradiation on soybean seeds promotes the recovery of soybean seedlings under oxidative stress, which positively regulates soybean growth through the regulation of glycolysis and redox related pathways.

Substrate Recognition by a Dual-Function P450 Monooxygenase GfsF Involved in FD-891 Biosynthesis.

GfsF is a multifunctional P450 monooxygenase that catalyzes epoxidation and subsequent hydroxylation in the biosynthesis of macrolide polyketide FD-891. Here, we describe the biochemical and structural analysis of GfsF. To obtain the structural basis of a dual-function reaction, we determined the crystal structure of ligand-free GfsF, which revealed GfsF to have a predominantly hydrophobic substrate binding pocket. The docking models, in conjunction with the results of the enzymatic assay with substrate analogues and site-directed mutagenesis suggested two distinct substrate binding modes for epoxidation and hydroxylation reactions, which explained how GfsF regulates the order of two oxidative reactions. These findings provide new insights into the reaction mechanism of multifunctional P450 monooxygenases.

Synthesis and biological activity of a novel fungicide, pyraziflumid.

Pyraziflumid was discovered as a novel SDHI fungicide chemically characterized by the 3-(trifluoromethyl)pyrazine-2-carboxamide group. This chemical series showed particularly high fungicidal activities against a broad spectrum of plant diseases in the case of N-(biphenyl-2-yl) as well as N-(1,1,3-trimethylindan-4-yl)carboxamides. Various N-(biphenyl-2-yl)pyrazine-2-carboxamides were synthesized, and their structure-activity relationships were studied. The optimization of the fungicidal performance of the series finally led to the identification of pyraziflumid, which could control a wide range of plant diseases. In this report, details of the structure-activity relationships from the lead compound to pyraziflumid are described.

Development of a novel acaricide, pyflubumide.

Pyflubumide is a novel carboxanilide acaricide discovered and developed by Nihon Nohyaku Co., Ltd., that exhibits excellent acaricidal activity against Tetranychus and Panonychus species, including strains that have developed resistance to conventional acaricides. Its safety profile against non-target arthropods is suitable for integrated pest management (IPM) programs. Pyflubumide was registered and launched in Japan in 2015 and Korea in 2017. This paper describes pyflubumide's invention history, synthesis, exploratory synthesis, biological activity, toxicological profile, and mode of action.

Gigahertz time-domain spectroscopy and imaging for non-destructive materials research and evaluation.

By using optical sampling with repetition frequency modulation of pump/probe laser pulses on photoconductive emitter/detector antennas, the high-speed time/frequency domain gigahertz imaging is reported due to the absence of opto-mechanical delay line in this optical scheme. The clear contrast for a 3-cm wide metal plate, which was placed behind a 5-cm thick concrete block, was observed with a 1 × 1 mm image pixilation. On average, it took only ~0.75 s per pixel/waveform acquisition/assignment with a 675 ps time-domain window. This could become a valuable non-destructive evaluation technique in gigahertz spectral range with all benefits of time-domain spectroscopy.

Parallel Post-Polyketide Synthase Modification Mechanism Involved in FD-891 Biosynthesis in Streptomyces graminofaciens A-8890.

To isolate a key polyketide biosynthetic intermediate for the 16-membered macrolide FD-891 (1), we inactivated two biosynthetic genes coding for post-polyketide synthase (PKS) modification enzymes: a methyltransferase (GfsG) and a cytochrome P450 (GfsF). Consequently, FD-892 (2), which lacks the epoxide moiety at C8-C9, the hydroxy group at C10, and the O-methyl group at O-25 of FD-891, was isolated from the gfsF/gfsG double-knockout mutant. In addition, 25-O-methyl-FD-892 (3) and 25-O-demethyl-FD-891 (4) were isolated from the gfsF and gfsG mutants, respectively. We also confirmed that GfsG efficiently catalyzes the methylation of 2 and 4 in vitro. Further, GfsF catalyzed the epoxidation of the double bond at C8-C9 of 2 and 3 and subsequent hydroxylation at C10, to afford 4 and 1, respectively. These results suggest that a parallel post-PKS modification mechanism is involved in FD-891 biosynthesis.

A matter of symmetry: terahertz polarization detection properties of a multi-contact photoconductive antenna evaluated by a response matrix analysis.

While terahertz time domain spectroscopy (THz-TDS) is a well-established technique, polarization sensitive measurements are challenging due to the need of broadband polarization devices. Here, we characterize our recently introduced multi-contact photoconductive detector antenna with a response matrix analysis. We show that the lead lines attached to electrodes reduce the antenna symmetry and thereby influence the properties of the response matrices. With a wire grid polarizer, we simulate a sample influencing the polarization angle and the intensity of the incident THz pulse. Evaluating the measurements with the response matrix analysis, our results show a well agreement of the adjusted and measured polarization angles and intensities over a frequency range from 0.25 to 0.8 THz.

Devising assisted reproductive technologies for wild-derived strains of mice: 37 strains from five subspecies of Mus musculus.

Wild-derived mice have long offered invaluable experimental models for mouse genetics because of their high evolutionary divergence from laboratory mice. A number of wild-derived strains are available from the RIKEN BioResource Center (BRC), but they have been maintained as living stocks because of the unavailability of assisted reproductive technology (ART). In this study, we sought to devise ART for 37 wild-derived strains from five subspecies of Mus musculus maintained at the BRC. Superovulation of females was effective (more than 15 oocytes per female) for 34 out of 37 strains by treatment with either equine chorionic gonadotropin or anti-inhibin serum, depending on their genetic background (subspecies). The collected oocytes could be fertilized in vitro at mean rates of 79.0% and 54.6% by the optimized protocol using fresh or frozen-thawed spermatozoa, respectively. They were cryopreserved at the 2-cell stage by vitrification with an ethylene glycol-based solution. In total, 94.6% of cryopreserved embryos survived the vitrification procedure and restored their normal morphology after warming. A conventional embryo transfer protocol could be applied to 25 out of the 35 strains tested. In the remaining 10 strains, live offspring could be obtained by a modified embryo transfer protocol using cyclosporin A treatment and co-transfer of ICR (laboratory mouse strain) embryos. Thus, ART for 37 wild-derived strains was devised successfully and is now routinely used for their preservation and transportation. The information provided here might facilitate broader use and wider distribution of wild-derived mice for biomedical research.

Tunneling dynamics and spectroscopic parameters of monodeuterated hydronium, H(2)DO(+), from a combined analysis of infrared and sub-millimeter spectra.

The infrared laser direct absorption spectrum of H(2)DO(+) in the OH stretching region was reported quite recently revealing large amplitude tunneling dynamics. The large rotational constants make the jet-cooled spectrum relatively sparse at low rotational temperatures and assignments thus challenging. Transitions were assigned through ground state combination differences, with additional tentative assignments made via comparison of predicted/observed spectra. More recently, 9 rotation-inversion transitions were recorded in the sub-millimeter (sub-mm) region, which yielded tunneling splittings and rotational constants differing slightly from IR results. This has prompted the present reinvestigation of the H(2)DO(+) spectra, which now takes full advantage of the combined data from both studies. While previous analyses considered each tunneling state as independent and non-interacting, the present analysis is based on a tunneling-Hamiltonian model for the well studied, isoelectronic NH(2)D molecule, modified to account for the larger tunneling splitting. The combined analysis revealed rotational interaction between tunneling states as well as between the two OH stretching modes and permitted a substantial number of new assignments to be made, including one sub-millimeter transition while only few IR assignments had to be corrected or omitted. It leads to improvement in parameters for both the ground as well as for the OH stretching states of this important molecular ion, which reproduce the assigned lines within experimental uncertainties, provides guidance, e.g., for the spectral search in the OD stretch region, and yields deeper insight into the tunneling dynamics.

Microwave spectrum of the HD2O+ ion: inversion-rotation transitions and inversion splitting.

Inversion-rotation spectral lines of the dideuterated hydronium ion, HD2O+, have been observed by a source-modulation millimeter- to submillimeter-wave spectrometer. The ion was generated by a hollow-cathode discharge in a gas mixture of D2O and H2O in a free-space cell. Ten inversion-rotation lines were measured precisely for the lowest pair of inversion doublets in the frequency region from 380 to 730 GHz. The observed lines include the most astronomically important transitions, 0(00) (-)-1(10)+ for the para species at 380 538.031(32) MHz and 1(01) (-)-1(11)+ for the ortho species at 728 420.189(34) MHz, which could be used as a radio astronomical probe investigating interstellar chemistry of deuterium fractionation. An analysis of the measured lines has yielded the rotational constants in the ground doublet states and the inversion splitting. The inversion splitting in the ground state was determined to be 808 866(34) MHz, that is, 26.980 87(113) cm(-1), where the numbers in parentheses give uncertainties estimated from the Jacobian matrix of the assumed centrifugal distortion constants. The determined inversion splitting is off by -0.51 cm(-1) from the predicted value of 27.49 cm(-1) by Rajamaki et al. using high-order coupled cluster ab initio calculation [J. Chem. Phys. 118, 10929 (2003)], and by -0.0510 cm(-1) from the observed value of 27.0318(72) cm(-1) by Dong et al. using high-resolution jet-cooled infrared spectroscopy [J. Chem. Phys. 122, 224301 (2005)] beyond the quoted uncertainty.

Microwave spectrum of the H2DO+ ion: inversion-rotation transitions and inversion splitting.

Inversion-rotation spectral lines of the monodeuterated hydronium ion, H(2)DO(+), have been observed by a source-modulation spectrometer in the millimeter- to submillimeter-wave region. The ion was generated by a hollow-cathode discharge in a gas mixture of H(2)O and D(2)O. Nine inversion-rotation lines were measured precisely for the lowest pair of inversion doublets in the frequency region from 210 to 720 GHz. The measured lines were analyzed to derive rotational constants in the inversion-doublet states and inversion splitting. The inversion splitting in the ground state was determined to be 1,215,866(410) MHz, that is, 40.5569(137) cm(-1), where the numbers in parentheses give probable uncertainties estimated from the Jacobian matrix of the assumed centrifugal distortion constants of the inversion-doublet states. The determined inversion splitting is off by -0.58 cm(-1) from the predicted value of 41.14 cm(-1) by Rayamaki et al. using high-order coupled cluster ab initio calculations [J. Chem. Phys. 118, 10929 (2003)], and by 0.039 cm(-1) from the observed value of 40.518(10) cm(-1) by Dong and Nesbitt using high-resolution jet-cooled infrared spectroscopy [J. Chem. Phys. 125, 144311 (2006)] beyond the quoted uncertainty. The most astronomically important transition 0(00)(-)-1(0)(+) for the ortho species was measured at 673,257.024(31) MHz, which could be used as a radioastronomical probe investigating interstellar chemistry of deuterium fractionation in space.

Basaloid carcinoma of the thymus: report of a case.

We report a case of basaloid carcinoma of the thymus, confirmed by histopathological examination, in a 52-year-old man who underwent complete resection. Local recurrence and lung metastases developed, which were obliterated by radiation therapy. The patient survived for 81 months after surgery. We reviewed the 13 other cases of basaloid carcinoma of the thymus reported in the literature. Presumably, this tumor is radiosensitive and its metastatic route is blood-borne.