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1.
Radiat Prot Dosimetry ; 198(17): 1283-1291, 2022 Oct 01.
Article in English | MEDLINE | ID: mdl-35876638

ABSTRACT

The crystal configuration dependence of thallium-doped caesium iodide (CsI(Tl)) scintillation detectors was analysed on an ambient dose equivalent (H*(10)). H*(10) were systematically calculated in various crystal conditions for aspect ratios and sizes in a virtual environment contaminated by radionuclides to investigate directional characteristics by comparing the H*(10)s with the typical irradiation geometries in anterior-posterior (AP), lateral (LAT), rotational (ROT) and isotropic (ISO). The simulation revealed that H*(10) obviously changes according to the crystal configuration and cuboidal CsI(Tl) scintillation detectors with specific aspect ratios could be applied to environmental dose rate measurement without further changes in the calibration procedure.


Subject(s)
Radioisotopes , Thallium , Calibration , Computer Simulation , Scintillation Counting
2.
Phys Rev Lett ; 126(21): 217202, 2021 May 28.
Article in English | MEDLINE | ID: mdl-34114835

ABSTRACT

CeIrSn with a quasikagome Ce lattice in the hexagonal basal plane is a strongly valence fluctuating compound, as we confirm by hard x-ray photoelectron spectroscopy and inelastic neutron scattering, with a high Kondo temperature of T_{K}∼480 K. We report a negative in-plane thermal expansion α/T below 2 K, which passes through a broad minimum near 0.75 K. Volume and a-axis magnetostriction for B∥a are markedly negative at low fields and change sign before a sharp metamagnetic anomaly at 6 T. These behaviors are unexpected for Ce-based intermediate valence systems, which should feature positive expansivity. Rather they point towards antiferromagnetic correlations at very low temperatures. This is supported by muon spin relaxation measurements down to 0.1 K, which provide microscopic evidence for a broad distribution of internal magnetic fields. Comparison with isostructural CeRhSn suggests that these antiferromagnetic correlations emerging at T≪T_{K} result from geometrical frustration.

3.
Radiat Prot Dosimetry ; 193(3-4): 228-236, 2021 May 17.
Article in English | MEDLINE | ID: mdl-33893735

ABSTRACT

Ambient dose equivalent rates were measured in the environment of the Fukushima prefecture using NaI(Tl)/CsI(Tl) scintillation detectors and CdZnTe/Ge semiconductor detectors. The dose rates obtained at the same locations varied beyond uncertainty (1σ). By replacing the spectrum-dose conversion operators obtained from the anterior-posterior geometry with those from the rotational geometry, the dose rates agreed with each other within uncertainties, except for a CsI(Tl) scintillation detector with a considerably flat crystal configuration, due to its excessive directional dependence.


Subject(s)
Air Pollutants, Radioactive , Radiation Monitoring , Radioactivity , Air Pollutants, Radioactive/analysis , Cesium , Semiconductors
4.
J Virol ; 94(15)2020 07 16.
Article in English | MEDLINE | ID: mdl-32461309

ABSTRACT

Oregano essential oil has long been known for its health-promoting benefits. Here, we report its activity against viral replication. Oregano oil was found to specifically inhibit lentiviruses, such as human and simian immunodeficiency viruses (HIV and SIV), irrespective of virus tropism, but not hepatitis C virus, adenovirus 5 (ADV5), Zika virus, and influenza (H1N1) virus. Oregano oil's most abundant components, carvacrol and its isomer, thymol, were shown to block virus-target cell fusion while not perturbing other stages of the virus life cycle. We detected changes in virus particle density, suggesting that cholesterol depletion from the HIV-1 envelope membrane reduces virus entry. Furthermore, infection was rescued by adding exogenous cholesterol. The evolution of viral resistance to carvacrol supported this mechanism of action with the identification of mutations in the viral gp41 fusion protein that counteracted cholesterol depletion. In addition, resistance to carvacrol emerged later than typically observed for other clinically used drugs, strengthening its antiviral potential. Structure-activity relationship studies revealed key motifs of carvacrol and thymol required for HIV neutralization and identified previously unknown active analogs. Carvacrol was also shown to additively cooperate with antiretroviral therapy. In sum, oregano oil and improved carvacrol and thymol analogs could be considered to supplement current HIV therapeutics.IMPORTANCE Oregano essential oil has multiple benefits in traditional medicine, cosmetics, and food industries. Carvacrol and its analog, thymol, are well-described components of oregano oil. Here, we show that these compounds inhibit HIV-target cell fusion independently of viral tropism. Our results suggest that carvacrol and thymol alter the cholesterol content of the viral membrane, blocking HIV-1 entry into the target cell. Resistance to carvacrol has selected for viruses with mutations in the viral envelope glycoprotein, gp41. This protein is known for its interaction with cholesterol present in membrane lipid rafts. Together, these results demonstrate the potential of therapies targeting the viral envelope membrane, and oregano oil is a safe supplement to antiretrovirals, potentially delaying disease progression and resistance development.


Subject(s)
Cymenes/pharmacology , HIV Envelope Protein gp41/metabolism , HIV-1/metabolism , Origanum/chemistry , Plant Oils/pharmacology , Virus Internalization/drug effects , Animals , Cholesterol/genetics , Cholesterol/metabolism , Cymenes/chemistry , Drug Resistance, Viral , HIV Envelope Protein gp41/genetics , HIV Infections/drug therapy , HIV Infections/genetics , HIV Infections/metabolism , HIV-1/genetics , HeLa Cells , Humans , Macaca mulatta , Mutation , Plant Oils/chemistry
5.
Retrovirology ; 15(1): 64, 2018 09 20.
Article in English | MEDLINE | ID: mdl-30236131

ABSTRACT

There is a constant need to improve antiretrovirals against HIV since therapy is limited by cost, side effects and the emergence of drug resistance. Kudzu is a climbing vine from which the root extract (Pueraria lobata), rich in isoflavones and saponins, has long been used in traditional Chinese medicine for a variety of purposes, from weight loss to alcoholism prevention. Here we show that Kudzu root extract significantly inhibits HIV-1 entry into cell lines, primary human CD4+T lymphocytes and macrophages, without cell-associated toxicity. Specifically, Kudzu inhibits the initial attachment of the viral particle to the cell surface, a mechanism that depends on the envelope glycoprotein gp120 but is independent from the HIV-1 cell receptor CD4 and co-receptors CXCR4/CCR5. This activity seems selective to lentiviruses since Kudzu inhibits HIV-2 and simian immunodeficiency virus, but does not interfere with Hepatitis C, Influenza, Zika Brazil and adenovirus infection. Importantly, depending on the dose, Kudzu can act synergistically or additively with the current antiretroviral cocktails against HIV-1 and can block   viruses resistant to the fusion inhibitor Enfuvirtide. Together our results highlight Kudzu's root extract value as a supplement to current antiretroviral therapy against HIV.


Subject(s)
Anti-HIV Agents/pharmacology , HIV-1/drug effects , Plant Extracts/pharmacology , Plant Roots/chemistry , Pueraria , Virus Attachment/drug effects , Animals , Cells, Cultured , Drug Synergism , Enfuvirtide , HIV Envelope Protein gp120/metabolism , HIV-1/physiology , Humans , Plant Extracts/chemistry , Virus Replication/drug effects
6.
Acta Physiol (Oxf) ; 222(2)2018 02.
Article in English | MEDLINE | ID: mdl-28786555

ABSTRACT

AIM: Stomach contractions show two types of specific patterns in many species, that is migrating motor contraction (MMC) and postprandial contractions (PPCs), in the fasting and fed states respectively. We found gastric PPCs terminated with migrating strong contractions in humans, dogs and suncus. In this study, we reveal the detailed characteristics and physiological implications of these strong contractions of PPC. METHODS: Human, suncus and canine gastric contractions were recorded with a motility-monitoring ingestible capsule and a strain-gauge force transducer. The response of motilin and ghrelin and its receptor antagonist on the contractions were studied by using free-moving suncus. RESULTS: Strong gastric contractions were observed at the end of a PPC in human, dog and suncus models, and we tentatively designated this contraction to be a postprandial giant contraction (PPGC). In the suncus, the PPGC showed the same property as those of a phase III contraction of MMC (PIII-MMC) in the duration, motility index and response to motilin or ghrelin antagonist administration. Ghrelin antagonist administration in the latter half of the PPC (LH-PPC) attenuated gastric contraction prolonged the duration of occurrence of PPGC, as found in PII-MMC. CONCLUSION: It is thought that the first half of the PPC changed to PII-MMC and then terminated with PIII-MMC, suggesting that PPC consists of a digestive phase (the first half of the PPC) and a discharge phase (LH-PPC) and that LH-PPC is coincident with MMC. In this study, we propose a new approach for the understanding of postprandial contractions.


Subject(s)
Gastrointestinal Motility/physiology , Ghrelin/metabolism , Motilin/metabolism , Postprandial Period/physiology , Shrews/physiology , Animals , Dogs , Humans , Muscle Contraction/physiology , Stomach/physiology
7.
J Chem Phys ; 147(2): 024501, 2017 Jul 14.
Article in English | MEDLINE | ID: mdl-28711051

ABSTRACT

In this paper, the nuclear quantum effect of the hydrogen molecule on its diffusivity was analyzed using the molecular dynamics (MD) method. The centroid MD (CMD) method was applied to reproduce the time evolution of the molecules. The diffusion coefficient of hydrogen was calculated using the Green-Kubo method over a wide temperature region, and the temperature dependence of the quantum effect of the hydrogen molecule on its diffusivity was addressed. The calculated results were compared with classical MD results based on the principle of corresponding state (PCS). It was confirmed that the difference in the diffusion coefficient calculated in the CMD and classical MD methods was small, and the PCS appears to be satisfied on the temperature dependence of the diffusion coefficient, even though the quantum effect of the hydrogen molecules was taken into account. It was clarified that this result did not suggest that the quantum effect on the diffusivity of the hydrogen molecule was small but that the two changes in the intermolecular interaction of hydrogen due to the quantum effect offset each other. Moreover, it was found that this tendency was related to the temperature dependence of the ratio of the kinetic energy of the quantum fluctuational motion to the classical kinetic energy.

8.
Radiat Prot Dosimetry ; 168(2): 190-6, 2016 Feb.
Article in English | MEDLINE | ID: mdl-25956785

ABSTRACT

The frequency distribution of the lineal energy, y, of a 30-MeV proton beam was measured as a function of the radial distance from the beam path, and the dosed mean of y, y¯(D), was obtained to investigate the radial dependence of y¯(D). A wall-less tissue-equivalent proportional counter, in a cylindrical volume with simulated diameters of 0.36, 0.72 and 1.44 µm was used for the measurement of y distributions, yf(y). The measured values of yf(y) summed in the radial direction agreed fairly well with the corresponding data taken from the microdosimetric calculations using the PHITS code. The y¯(D) value of the 30-MeV proton beam presented its smallest value at r = 0.0 and gradually increased with radial distance, and the y¯(D) values of heavy ions such as iron showed rapid decrease with radial distance. This experimental result demonstrated that the stochastic deposited energy distribution of high-energy protons in the microscopic region is rather constant in the core as well as in the penumbra region of the track structure.


Subject(s)
Computer Simulation , Heavy Ions , Linear Energy Transfer , Phantoms, Imaging , Protons , Radiation Monitoring/instrumentation , Radiation Monitoring/methods , Humans , Radiation Dosage
9.
Phys Rev Lett ; 115(25): 256405, 2015 Dec 18.
Article in English | MEDLINE | ID: mdl-26722935

ABSTRACT

We study the electronic structure of bulk single crystals and epitaxial films of Fe_{3}O_{4}. Fe 2p core level spectra show clear differences between hard x-ray (HAX) and soft x-ray photoemission spectroscopy (PES). The bulk-sensitive spectra exhibit temperature (T) dependence across the Verwey transition, which is missing in the surface-sensitive spectra. By using an extended impurity Anderson full-multiplet model-and in contrast to an earlier peak assignment-we show that the two distinct Fe species (A and B site) and the charge modulation at the B site are responsible for the newly found double peaks in the main peak above T_{V} and its T-dependent evolution. The Fe 2p HAXPES spectra show a clear magnetic circular dichroism (MCD) in the metallic phase of magnetized 100-nm-thick films. The model calculations also reproduce the MCD and identify the contributions from magnetically distinct A and B sites. Valence band HAXPES shows a finite density of states at E_{F} for the polaronic half metal with a remnant order above T_{V} and a clear gap formation below T_{V}. The results indicate that the Verwey transition is driven by changes in the strongly correlated and magnetically active B-site electronic states, consistent with resistivity and optical spectra.

10.
J Environ Radioact ; 139: 260-265, 2015 Jan.
Article in English | MEDLINE | ID: mdl-24698118

ABSTRACT

The car-borne survey system KURAMA-II, developed by the Kyoto University Research Reactor Institute, has been used for air dose rate mapping after the Fukushima Dai-ichi Nuclear Power Plant accident. KURAMA-II consists of a CsI(Tl) scintillation detector, a GPS device, and a control device for data processing. The dose rates monitored by KURAMA-II are based on the G(E) function (spectrum-dose conversion operator), which can precisely calculate dose rates from measured pulse-height distribution even if the energy spectrum changes significantly. The characteristics of KURAMA-II have been investigated with particular consideration to the reliability of the calculated G(E) function, dose rate dependence, statistical fluctuation, angular dependence, and energy dependence. The results indicate that 100 units of KURAMA-II systems have acceptable quality for mass monitoring of dose rates in the environment.


Subject(s)
Air Pollutants, Radioactive/analysis , Fukushima Nuclear Accident , Radiation Monitoring/methods , Radioactive Fallout/analysis , Radioisotopes/analysis , Automobiles , Japan , Reproducibility of Results
11.
Rev Sci Instrum ; 85(11): 11D813, 2014 Nov.
Article in English | MEDLINE | ID: mdl-25430226

ABSTRACT

A Rogowski probe consisting of a small multi-layer Rogowski coil, five magnetic pick-up coils, and a Langmuir probe was developed to measure the local current density and its direction. It can be moved along the major radius and can be turned around its axis. This probe was used to measure the current density profile near the last closed flux surface of Ohmic plasmas in Tokyo Spherical Tokamak-2. The current density profile was measured successfully with a signal to noise ratio of greater than 20.

12.
Rev Sci Instrum ; 85(11): 11D846, 2014 Nov.
Article in English | MEDLINE | ID: mdl-25430259

ABSTRACT

The multi-pass Thomson scattering (TS) scheme enables obtaining many photons by accumulating multiple TS signals. The signal-to-noise ratio (SNR) depends on the accumulation number. In this study, we performed multi-pass TS measurements for ohmically heated plasmas, and the relationship between SNR and the accumulation number was investigated. As a result, improvement of SNR in this experiment indicated similar tendency to that calculated for the background noise dominant situation.

13.
Rev Sci Instrum ; 85(5): 056103, 2014 May.
Article in English | MEDLINE | ID: mdl-24880428

ABSTRACT

In multi-pass Thomson scattering (TS) scheme, a laser pulse makes multiple round trips through the plasma, and the effective laser energy is enhanced, and we can increase the signal-to-noise ratio as a result. We have developed a coaxial optical cavity in which a laser pulse is confined, and we performed TS measurements using the coaxial cavity in tokamak plasmas for the first time. In the optical cavity, the laser energy attenuation was approximately 30% in each round trip, and we achieved a photon number gain of about 3 compared with that obtained in the first round trip. In addition, the temperature measurement accuracy was improved by accumulating the first three round trip waveforms.

14.
J Chem Phys ; 140(13): 134506, 2014 Apr 07.
Article in English | MEDLINE | ID: mdl-24712800

ABSTRACT

In this paper, we describe the analysis of the thermodynamic properties of cryogenic hydrogen using classical molecular dynamics (MD) and path integral MD (PIMD) method to understand the effects of the quantum nature of hydrogen molecules. We performed constant NVE MD simulations across a wide density-temperature region to establish an equation of state (EOS). Moreover, the quantum effect on the difference of molecular mechanism of pressure-volume-temperature relationship was addressed. The EOS was derived based on the classical mechanism idea only using the MD simulation results. Simulation results were compared with each MD method and experimental data. As a result, it was confirmed that although the EOS on the basis of classical MD cannot reproduce the experimental data of saturation property of hydrogen in the high-density region, the EOS on the basis of PIMD well reproduces those thermodynamic properties of hydrogen. Moreover, it was clarified that taking quantum effects into account makes the repulsion force larger and the potential well shallower. Because of this mechanism, the intermolecular interaction of hydrogen molecules diminishes and the virial pressure increases.

15.
Plant Dis ; 98(5): 698, 2014 May.
Article in English | MEDLINE | ID: mdl-30708510

ABSTRACT

In 2009, some commercially grown tomato (Solanum lycopersicum) plants in Chiba Prefecture, Japan, exhibited mosaic symptoms. Ten plants from a total of about 72,000 cultivated plants in the greenhouses showed such symptoms. To identify the causal agent, sap from leaves of the diseased plants was inoculated into Chenopodium quinoa and Nicotiana benthamiana plants. Local necrotic lesions appeared on inoculated leaves of C. quinoa, but no systemic infection was observed. Systemic mosaic symptoms were observed on the N. benthamiana plants inoculated. Single local lesion isolation was performed three times using C. quinoa to obtain a reference isolate for further characterization. N. benthamiana was used for propagation of the isolate. Sap from infected leaves of N. benthamiana was mechanically inoculated into three individual S. lycopersicum cv. Momotaro. Symptoms appearing on inoculated tomatoes were indistinguishable from those of diseased tomato plants found initially in the greenhouse. Flexuous, filamentous particles, ~750 nm long, were observed by electron microscopy in the sap of the tomato plants inoculated with the isolate, indicating that the infecting virus may belong to the family Potyviridae. To determine genomic sequence of the virus, RT-PCR was performed. Total RNA was extracted from the tomato leaves experimentally infected with the isolate using an RNeasy Plant Mini kit (QIAGEN, Hilden, Germany). RT-PCR was performed by using a set of universal, degenerate primers for Potyviruses as previously reported (2). Amplicons (~1,500 bp) generated by RT-PCR were extracted from the gels using the QIAquick Gel Extraction kit (QIAGEN) and cloned into pCR-BluntII TOPO (Invitrogen, San Diego, CA). DNA sequences of three individual clones were determined using a combination of plasmid and virus-specific primers, showing that identity among three clones was 99.8%. A consensus nucleotide sequence of the isolate was deposited in GenBank (AB823816). BLASTn analysis of the nucleotide sequence determined showed 99% identity with a partial sequence in the NIb/coat protein (CP) region of Colombian datura virus (CDV) tobacco isolate (JQ801448). Comparison of the amino acid sequence predicted for the CP with previously reported sequences for CDV (AY621656, AJ237923, EU571230, AM113759, AM113754, and AM113761) showed 97 to 100% identity range. Subsequently, CDV infection in both the original and experimentally inoculated plants was confirmed by RT-PCR using CDV-specific primers (CDVv and CDVvc; [1]), and, hence, the causal agent of the tomato disease observed in greenhouse tomatoes was proved to be CDV. The first case of CDV on tomato was reported in Netherlands (3), indicating that CDV was transmitted by aphids from CDV-infected Brugmansia plants cultivated in the same greenhouse. We carefully investigated whether Brugmansia plants naturally grew around the greenhouses, but we could not find them inside or in proximity to the greenhouses. Therefore, sources of CDV inoculum in Japan are still unclear. This is the first report of a mosaic disease caused by CDV on commercially cultivated S. lycopersicum in Japan. References: (1) D. O. Chellemi et al. Plant Dis. 95:755, 2011. (2) J. Chen et al. Arch. Virol. 146:757, 2001. (3) J. Th. J. Verhoeven et al. Eur. J. Plant. Pathol. 102:895, 1996.

16.
Food Chem ; 127(3): 1114-8, 2011 Aug 01.
Article in English | MEDLINE | ID: mdl-25214103

ABSTRACT

The aim of this study was to evaluate the use of vacuum-impregnation (VI) for enriching the ascorbic acid content of whole potatoes. Whole potatoes were immersed in a 10% ascorbic acid (AA) solution. A vacuum pressure of 70cm Hg was applied for 0-60min, following atmospheric pressure restoration for 3h, while samples remained in the VI solution. AA concentrations of potatoes were measured using HPLC. The effects of cooking and storage time in subsets of the fortified samples were also evaluated. Results indicated that the AA concentration of whole potatoes increased with vacuum time (max 150mg/100g fr. wt.). In addition, a steam-cooking study showed that 100g of the 25min steam-cooked VI potatoes could provide adults with 90-100% of the recommended daily allowance of AA (100mg). The storage study showed that VI whole potatoes had a relatively high AA concentration (50mg/100gfr. wt.), even at 14days of storage at 4°C. This study indicated that VI treatment of whole potatoes was useful for enriching the AA content.

17.
Radiat Prot Dosimetry ; 143(2-4): 450-4, 2011 Feb.
Article in English | MEDLINE | ID: mdl-21183535

ABSTRACT

Effects of structural materials in a wall-less tissue-equivalent proportional counter were evaluated based on the calculation of energy deposits by EGS5 and the measurement of lineal energy distributions using 290 MeV u(-1) carbon beams. It is found that the correction of measured data based on simulation is necessary for understanding the energy deposition spectra in the homogeneous condition in tissues.


Subject(s)
Biomimetic Materials , Carbon , Heavy Ions , Radiation Monitoring/instrumentation , Equipment Design , Equipment Failure Analysis , Radiation Dosage
18.
Phys Rev Lett ; 104(10): 106401, 2010 Mar 12.
Article in English | MEDLINE | ID: mdl-20366439

ABSTRACT

The Magnéli phase Ti(4)O(7) exhibits two sharp jumps in resistivity with coupled structural transitions as a function of temperature at T(c1) approximately 142 K and T(c2) = 154 K. We have studied electronic structure changes across the two transitions using 7 eV laser, soft x-ray, and hard x-ray (HX) photoemission spectroscopy (PES). Ti 2p-3d resonant PES and HX PES show a clear metallic Fermi edge and mixed valency above T(c2). The low temperature phase below T(c1) shows a clear insulating gap of approximately 100 meV. The intermediate phase between T(c1) and T(c2) indicates a pseudogap coexisting with remnant coherent states. HX PES and complementary calculations have confirmed the coherent screening in the strongly correlated intermediate phase. The results suggest the existence of a highly anomalous state sandwiched between the mixed-valent Fermi liquid and charge ordered Mott-insulating phase in Ti(4)O(7).

19.
J Phys Condens Matter ; 22(9): 095502, 2010 Mar 10.
Article in English | MEDLINE | ID: mdl-21389417

ABSTRACT

CeOs(4)Sb(12) and CeFe(4)P(12) are classified as Kondo semiconductors, which show coupled changes in electrical transport, thermodynamic and magnetic properties with a low-temperature semiconductor-like electrical resistivity. We have carried out core level and valence band photoemission spectroscopy on single crystal CeOs(4)Sb(12) and CeFe(4)P(12) to study their electronic structure and the evolution of states at the Fermi level as a function of temperature (∼10-300 K). The Ce 3d core level spectra show the presence of f(0), f(1) and f(2) final states with very different relative intensities in the two compounds. Single-impurity Anderson model calculations provide f electron counts of n(f) = 0.97 and 0.86 per Ce atom, suggestive of a low- and high-T(K) (= single ion Kondo temperature) for CeOs(4)Sb(12) and CeFe(4)P(12), respectively. The high-resolution temperature-dependent near-Fermi level spectra show pseudogaps of energy ∼ 50 meV and ∼ 110 meV in the valence band density of states (DOS) of CeOs(4)Sb(12) and CeFe(4)P(12), respectively. The temperature dependence of the DOS at the Fermi level follows the change in effective magnetic moment estimated from magnetic susceptibility for both materials, confirming the Kondo nature of the pseudogap in CeOs(4)Sb(12) and CeFe(4)P(12). A compilation of measured pseudogaps using photoemission and optical spectroscopy identifies the charge gaps Δ(C) for Ce-based Kondo semiconductors and provides a direct relation with T(K) given by Δ(C) ∼ 2k(B)T(K). In conjunction with the known behaviour of the spin gaps Δ(S) ∼ k(B)T(K), the results establish the coupled energy scaling of the spin and charge gaps in Kondo semiconductors.

20.
J Anim Sci ; 87(10): 3134-41, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19542507

ABSTRACT

Skeletal muscle growth is accomplished chiefly through the actions of satellite cells, a heterogeneous population that includes the adult muscle stem cell. Located adjacent to a mature muscle fiber, satellite cells typically reside in a quiescent state. Little information exists detailing satellite cell regulation of reversible G(0). One member of the mitosin family of centromere proteins, LEK1 (leucine/glutamic acid/lysine protein 1), is present in the nucleus of nondividing mouse satellite cells. The objective of this study was to evaluate LEK1 as a marker of quiescent bovine satellite cells (BSC) in vitro and in vivo. The BSC were isolated from young bull calves (< or =7 d) and cultured in vitro for up to 9 d before fixation and immunostaining for LEK1. Results demonstrated that all myogenic cells contain the protein, with immunostaining primarily within the nucleus and immediate perinuclear region. Immunocytochemical detection of LEK1 in cryosections of mature cows revealed that the protein was present in a fraction of satellite cells and muscle fiber nuclei. Approximately 20% of Pax7-expressing satellite cells contained LEK1. An equivalent percentage of myonuclei, as defined by nuclei within a dystrophin boundary, contained nuclear LEK1. To gain insight into the functional role of LEK1, BSC were transiently transfected with plasmids coding for putative dominant inhibitory LEK1 proteins [DeltaLEK1(991) and DeltaLEK1(911)] and evaluated for cell proliferation. Both forms of DeltaLEK1 inhibited (P < 0.05) BSC proliferation, as indicated by a decrease in Ki67 immunopositive cells. In C2C12 myoblasts, DeltaLEK1(911) inhibited (P < 0.05) myoblast determination protein 1 (MyoD)-directed muscle gene transcriptional activity; DeltaLEK1(991) had no effect on TnI-Luc transcription. By contrast, both DeltaLEK1 fusion proteins inhibited myogenin expression in BSC without disrupting myoblast fusion. These results provide evidence that LEK1 serves to coordinate proliferation and differentiation in myogenic cells. Coupling the immunostaining pattern and functional data, we propose that LEK1 may serve as a useful marker for satellite cells that are preparing to fuse into adjacent fibers as well as an indicator of recently added myonuclei.


Subject(s)
Cattle/physiology , Chromosomal Proteins, Non-Histone/physiology , Muscle Development/physiology , Muscle Fibers, Skeletal/physiology , Satellite Cells, Skeletal Muscle/physiology , Animals , Blotting, Western/veterinary , Cell Proliferation , Female , Immunohistochemistry/veterinary , Male , Muscle Fibers, Skeletal/cytology , Mutagenesis , Plasmids/pharmacology , Satellite Cells, Skeletal Muscle/cytology , Transfection/veterinary
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