Development of a method to rapidly assess resistance/susceptibility of Micro-Tom tomatoes to Tomato yellow leaf curl virus via agroinoculation of cotyledons

Objective: Tomato yellow leaf curl virus (TYLCV) is one of the pathogens severely damaging tomato crops. Therefore, methods to treat or prevent TYLCV infection need to be developed. For this purpose, a method to conveniently and quickly assess infection of tomatoes by TYLCV is desired. In the present study, we established a quick method to evaluate TYLCV infection using cotyledons of Micro-Tom, a miniature tomato cultivar. Results: First, we constructed a binary plasmid harboring 1.5 copies of the TYLCV genome and transformed Agrobacterium with the plasmid. By injecting agroinoculum from the resulting transformant into the branches of Micro-Tom, we confirmed the susceptibility of Micro-Tom to TYLCV. To shorten the evaluation process of TYLCV infection further, we agroinoculated cotyledons of Micro-Tom 10 days after sowing seeds. We consistently observed typical symptoms of TYLCV infection on true leaves 10 days after agroinoculation. Molecular analysis detected TYLCV progeny DNA in all leaves demonstrating symptoms 6 days after agroinoculation. Therefore, our new protocol enabled assessment of TYLCV infection within 20 days after sowing seeds. Thus, agroinoculation of Micro-Tom cotyledons will accelerate the process of screening TYLCV-resistant Micro-Toms and enable screening of larger numbers of plants more quickly, contributing to the development of TYLCV-resistant tomatoes

Ultrafast Feed Drilling of Carbon Fiber-Reinforced Thermoplastics

Carbon fiber-reinforced thermoplastics (CFRTP) are just beginning to be utilized for various applications such as aerospace, automobiles, and sporting goods in place of CFRPs, and the demand for through-hole drilling of CFRTPs is increasing. In this study, the machinability in drilling of CFRTPs under various conditions was experimentally analyzed in terms of the material properties, and a feasibility study of ultrafast feed drilling was conducted. The results showed that delamination at the outlet surface can be significantly suppressed during high rotational drilling when the feed rate is set to more than 3000 mm/min. By providing appropriate drilling conditions to prevent polymers in CFRTPs from softening, ultra-fast drilling of CFRTPs was successfully achieved under dry conditions

Graphical Evaluation Method for Void Distribution in Direct Energy Deposition

AbstractDirect energy deposition (DED) process attracts attention from industries because of its applicability to production of complex shape parts. However, technical challenges still remain in DED, such as void evolution inside of the produced object. This paper provides a new graphical evaluation method of void distribution. Binarizing a cross-sectional image of the cladded object and applying a 2-dimensional Gaussian window, the void distribution rate is separately evaluated in each local area on the cross section. To clarify the relation between void evolution and deposition condition in Inconel 625, the void distribution is experimentally evaluated through the various tests

13C/15N‐Enriched L‐Dopa as a Triple‐Resonance NMR Probe to Monitor Neurotransmitter Dopamine in the Brain and Liver Extracts of Mice

In an attempt to monitor μM-level trace constituents, we applied here 1H-{13C-15N} triple-resonance nuclear magnetic resonance (NMR) to 13C/15N-enriched L-Dopa as the inevitable precursor of the neurotransmitter dopamine in the brain. The perfect selectivity (to render endogenous components silent) and μM-level sensitivity (700 MHz spectrometer equipped with a cryogenic probe) of triple-resonance allowed the unambiguous and quantitative metabolic and pharmacokinetic analyses of administered L-Dopa/dopamine in the brain and liver of mice. The level of dopamine generated in the brain (within the range 7–76 μM, which covers the typical stimulated level of ~30 μM) could be clearly monitored ex vivo, but was slightly short of the detection limit of a 7T MR machine for small animals. This work suggests that μM-level trace constituents are potential targets of ex vivo monitoring as long as they contain N atom(s) and their appropriate 13C/15N-enrichment is synthetically accessible

Photoacoustic in vivo 3D imaging of tumor using a highly tumor-targeting probe under high-threshold conditions

Three-dimensional (3D) representation of a tumor with respect to its size, shape, location, and boundaries is still a challenge in photoacoustic (PA) imaging using artificial contrast agents as probes. We carried out PA imaging of tumors in mice using 800RS-PMPC, which was obtained by coupling of 800RS, a near-infrared cyanine dye, with PMPC, a highly selective tumor-targeting methacrylate polymer having phosphorylcholine side chains, as a probe. The conjugate 800RS-PMPC forms compact nanoparticles (dDLS = 14.3 nm), retains the biocompatibility of the parent polymer (PMPC) and exhibits unprecedented PA performance. When applied to mice bearing a 6 × 3 × 3 mm3 tumor buried 6 mm beneath the skin, the probe 800RS-PMPC selectively accumulates in the tumor and emits PA signals that are strong enough to be unambiguously distinguished from noise signals of endogenous blood/hemoglobin. The PA image thus obtained under high-threshold conditions allows 3D characterization of the tumor in terms of its size, shape, location, and boundaries

Anisotropic transport in unidirectional lateral superlattice around half-filling of the second Landau level

We have observed marked transport anisotropy in short period (a=92 nm) unidirectional lateral superlattices around filling factors nu=5/2 and 7/2: magnetoresistance shows a sharp peak for current along the modulation grating while a dip appears for current across the grating. By altering the ratio a/l (with l=sqrt{hbar/eB_perp} the magnetic length) via changing the electron density n_e, it is shown that the nu=5/2 anisotropic features appear in the range 6.6 alt a/l alt 7.2 varying their intensities, becoming most conspicuous at a/l simeq 6.7. The peak/dip broadens with temperature roughly preserving its height/depth up to 250 mK. Tilt experiments reveal that the structures are slightly enhanced by an in-plane magnetic field B_| perpendicular to the grating but are almost completely destroyed by B_| parallel to the grating. The observations suggest the stabilization of a unidirectional charge-density-wave or stripe phase by weak external periodic modulation at the second Landau level.Comment: REVTeX, 5 pages, 3 figures, Some minor revisions, Added notes and reference

Heart Failure-Inducible Gene Therapy Targeting Protein Phosphatase 1 Prevents Progressive Left Ventricular Remodeling

BACKGROUND: The targeting of Ca(2+) cycling has emerged as a potential therapy for the treatment of severe heart failure. These approaches include gene therapy directed at overexpressing sarcoplasmic reticulum (SR) Ca(2+) ATPase, or ablation of phospholamban (PLN) and associated protein phosphatase 1 (PP1) protein complexes. We previously reported that PP1β, one of the PP1 catalytic subunits, predominantly suppresses Ca(2+) uptake in the SR among the three PP1 isoforms, thereby contributing to Ca(2+) downregulation in failing hearts. In the present study, we investigated whether heart-failure-inducible PP1β-inhibition by adeno-associated viral-9 (AAV9) vector mediated gene therapy is beneficial for preventing disease progression in genetic cardiomyopathic mice. METHODS: We created an adeno-associated virus 9 (AAV9) vector encoding PP1β short-hairpin RNA (shRNA) or negative control (NC) shRNA. A heart failure inducible gene expression system was employed using the B-type natriuretic protein (BNP) promoter conjugated to emerald-green fluorescence protein (EmGFP) and the shRNA sequence. AAV9 vectors (AAV9-BNP-EmGFP-PP1βshRNA and AAV9-BNP-EmGFP-NCshRNA) were injected into the tail vein (2×10(11) GC/mouse) of muscle LIM protein deficient mice (MLPKO), followed by serial analysis of echocardiography, hemodynamic measurement, biochemical and histological analysis at 3 months. RESULTS: In the MLPKO mice, BNP promoter activity was shown to be increased by detecting both EmGFP expression and the induced reduction of PP1β by 25% in the myocardium. Inducible PP1βshRNA delivery preferentially ameliorated left ventricular diastolic function and mitigated adverse ventricular remodeling. PLN phosphorylation was significantly augmented in the AAV9-BNP-EmGFP-PP1βshRNA injected hearts compared with the AAV9-BNP-EmGFP-NCshRNA group. Furthermore, BNP production was reduced, and cardiac interstitial fibrosis was abrogated at 3 months. CONCLUSION: Heart failure-inducible molecular targeting of PP1β has potential as a novel therapeutic strategy for heart failure