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•Significant, 3–4 fold sensitivity gains are obtained experiments with a novel CPMAS CryoProbe.•The benefits are demonstrated in 2D and 3D experiments on five challenging biological ...assemblies.•High-quality 2D NN and NCA/NCO spectra are obtained on a sample with carbons at natural abundance.
Despite breakthroughs in MAS NMR hardware and experimental methodologies, sensitivity remains a major challenge for large and complex biological systems. Here, we report that 3–4 fold higher sensitivities can be obtained in heteronuclear-detected experiments, using a novel HCN CPMAS probe, where the sample coil and the electronics operate at cryogenic temperatures, while the sample is maintained at ambient temperatures (BioSolids CryoProbe™). Such intensity enhancements permit recording 2D and 3D experiments that are otherwise time-prohibitive, such as 2D 15N-15N proton-driven spin diffusion and 15N-13C double cross polarization to natural abundance carbon experiments. The benefits of CPMAS CryoProbe-based experiments are illustrated for assemblies of kinesin Kif5b with microtubules, HIV-1 capsid protein assemblies, and fibrils of human Y145Stop and fungal HET-s prion proteins – demanding systems for conventional MAS solid-state NMR and excellent reference systems in terms of spectral quality. We envision that this probe technology will be beneficial for a wide range of applications, especially for biological systems suffering from low intrinsic sensitivity and at physiological temperatures.
A new scanning system named “Vertex picker” has been developed to rapid collect alpha decay events, which are calibration sources for the range-energy relation in nuclear emulsion. A ...computer-controlled optical microscope scans emulsion layers exhaustively, and a high-speed and high-resolution camera takes their micrographs. A dedicated image processing picks out vertex-like shapes. Practical operations of alpha decay search were demonstrated by emulsion sheets of the KEK-PS E373 experiment. Alpha decays of nearly 28 events were detected in eye-check work on a PC monitor per hour. This yield is nearly 20 times more effective than that by the conventional eye-scan method. The speed and quality is acceptable for the coming new experiment, J-PARC E07.
A laboratory prototype system that correlates murine blood absorbance with degree of infection for Plasmodium berghei and Trypanosoma avensi has been designed, constructed and tested. A population ...(n = 6) of control uninfected, Plasmodium infected and Trypanosoma infected BALB/c mice were developed and spectral absorption measurements pre and post infection were made every 3 days. A fibre optic spectrometer set-up was used as the basis of a laboratory prototype biosensor that uses the Beer Lambert Law to relate Ultraviolet–Visible–Near-infrared absorbance data to changes in murine blood chemistry post infection. Spectral absorption results indicate a statistically relevant correlation at a 650 nm with infection for Plasmodium from between 4 and 7 sampling days' post infection, in spite of significant standard deviations among the sample populations for control and infected mice. No significant spectral absorption change for Trypanosoma infection was been detected from the current data. Corresponding stained slides of control and infected blood at each sampling date were taken with related infected cell counts determined and these correlate well for Plasmodium absorbance at 650 nm.
•A laboratory prototype system to determine murine blood UV-Visible-Near-Infrared absorbance has been devel with degree of infection for Plasmodium berghei and Trypanosoma avensi has been designed, constructed and tested.•Spectral absorption results indicate a statistically relevant correlation at a 650 nm with infection for Plasmodium.•No significant spectral absorption change for Trypanosoma infection was been detected from the current data.
Abstract
Ξ− atomic X-ray spectroscopy is one of the most useful methods for investigation of the Ξ–nucleus strong interaction. Since the X-ray energy is shifted and/or broadened due to the Ξ–nucleus ...strong interaction compared to those calculated from electromagnetic interaction alone, the measurement of the energy shift, ΔE, and the width, Γ, give us information on the Ξ–nucleus potential. A serious problem in the measurement is the significant background derived from in-flight Ξ− decay. A novel method of identifying stopped Ξ− events using the nuclear emulsion was developed to realize the first Ξ− atomic X-ray spectroscopy experiment as the J-PARC E07 experiment, which also aimed at searching for ΛΛ and Ξ− hypernuclei in the emulsion. The X-rays emitted from Ξ− Br and Ξ− Ag atoms were measured using germanium detectors. No clear peaks were observed in the obtained spectra. However, we succeeded in reducing the background to 1/170 by this method employing coincidence measurements using nuclear emulsion and X-ray detectors.
Corneal opacification due to fibrosis is a leading cause of blindness worldwide. Fibrosis occurs from many causes including trauma, photorefractive surgery, microbial keratitis (infection of the ...cornea), and chemical burns, yet there is a paucity of therapeutics to prevent or treat corneal fibrosis. This study aimed to determine if andrographolide, a labdane diterpenoid found in
, has anti-fibrotic properties. Furthermore, we evaluated if andrographolide could prevent the differentiation of fibroblasts to myofibroblasts
, given that the transforming growth factor beta-1(TGF-β1) stimulated persistence of myofibroblasts in the cornea is a primary component of fibrosis. We demonstrated that andrographolide inhibited the upregulation of alpha smooth muscle actin (αSMA) mRNA and protein in rabbit corneal fibroblasts (RCFs), thus, demonstrating a reduction in the transdifferentiation of myofibroblasts. Immunofluorescent staining of TGF-β1-stimulated RCFs confirmed a dose-dependent decrease in αSMA expression when treated with andrographolide. Additionally, andrographolide was well tolerated
and had no impact on corneal epithelialization in a rat debridement model. These data support future studies investigating the use of andrographolide as an anti-fibrotic in corneal wound healing.
Abstract
A double-$\Lambda$ hypernucleus, _{\Lambda\Lambda}\mathrm{Be}$, was observed by the J-PARC E07 Collaboration in nuclear emulsions tagged by the ($K^{-}, K^{+}$) reaction. This event was ...interpreted as the production and decay of $ {}_{\Lambda\Lambda}^{\;10}\mathrm{Be}$, _{\Lambda\Lambda}^{\;11}\mathrm{Be}$, or _{\Lambda\Lambda}^{\;12}\mathrm{Be}^{*}$ via $\Xi^{-}$ capture in ^{16}\mathrm{O}$. By assuming capture in the atomic 3D state, the binding energies of two $\Lambda$ hyperons ($B_{\Lambda\Lambda}$) of these double-$\Lambda$ hypernuclei are obtained to be $15.05 \pm 0.11\,\mathrm{MeV}$, $19.07 \pm 0.11\,\mathrm{MeV}$, and $13.68 \pm 0.11\,\mathrm{MeV}$, respectively. Based on the kinematic fitting, _{\Lambda\Lambda}^{\;11}\mathrm{Be}$ is the most likely explanation for the observed event.
Corneal opacification due to fibrosis is a leading cause of blindness worldwide. Fibrosis occurs from many causes including trauma, photorefractive surgery, microbial keratitis (infection of the ...cornea), and chemical burns, yet there is a paucity of therapeutics to prevent or treat corneal fibrosis. This study aimed to determine if andrographolide, a labdane diterpenoid found in Andrographis paniculate, has anti-fibrotic properties. Furthermore, we evaluated if andrographolide could prevent the differentiation of fibroblasts to myofibroblasts in vitro, given that the transforming growth factor beta-1(TGF-β1) stimulated persistence of myofibroblasts in the cornea is a primary component of fibrosis. We demonstrated that andrographolide inhibited the upregulation of alpha smooth muscle actin (αSMA) mRNA and protein in rabbit corneal fibroblasts (RCFs), thus, demonstrating a reduction in the transdifferentiation of myofibroblasts. Immunofluorescent staining of TGF-β1-stimulated RCFs confirmed a dose-dependent decrease in αSMA expression when treated with andrographolide. Additionally, andrographolide was well tolerated in vivo and had no impact on corneal epithelialization in a rat debridement model. These data support future studies investigating the use of andrographolide as an anti-fibrotic in corneal wound healing.
Abstract
Bound systems of $\Xi^-$–$^{14}_{}{\rm N}$ are studied via $\Xi^-$ capture at rest followed by emission of a twin single-$\Lambda$ hypernucleus in the emulsion detectors. Two events forming ...extremely deep $\Xi^-$ bound states were obtained by analysis of a hybrid method in the E07 experiment at J-PARC and reanalysis of the E373 experiment at KEK-PS. The decay mode of one event was assigned as $\Xi^-+^{14}\!\,\!\!{\rm N}\to^{5}_{\Lambda}\!\!\!{\rm He} + ^{5}_{\Lambda}\!\!{\rm He} + ^{4}\!\!{\rm He} + \mathrm{n}$. Since there are no excited states for daughter particles, the binding energy of the $\Xi^-$ hyperon, $B_{\Xi^-}$, in the $^{14}_{}{\rm N}$ nucleus was uniquely determined to be $6.27 \pm 0.27$ MeV. Another $\Xi^-$–$^{14}_{}{\rm N}$ system via the decay $^{9}_{\Lambda}{\rm Be} + ^{5}_{\Lambda}\!\!\!{\rm He} + \mathrm{n}$ brings a $B_{\Xi^-}$ value, $8.00 \pm 0.77$ MeV or $4.96 \pm 0.77$ MeV, where the two possible values of $B_{\Xi^-}$ correspond to the ground and the excited states of the daughter $^{9}_{\Lambda}{\rm Be}$ nucleus, respectively. Because the $B_{\Xi^-}$ values are larger than those of the previously reported events (KISO and IBUKI), which are both interpreted as the nuclear $1p$ state of the $\Xi^-$–$^{14}_{}{\rm N}$ system, these new events give the first indication of the nuclear $1s$ state of the $\Xi$ hypernucleus, $^{15}_{\Xi}{\rm C}$.
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