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1.
J Synchrotron Radiat ; 28(Pt 3): 977-986, 2021 May 01.
Article in English | MEDLINE | ID: mdl-33950006

ABSTRACT

We report on the development of a high-resolution and highly efficient beamline for soft X-ray resonant inelastic X-ray scattering (RIXS) located at the Taiwan Photon Source. This beamline adopts an optical design that uses an active grating monochromator (AGM) and an active grating spectrometer (AGS) to implement the energy compensation principle of grating dispersion. Active gratings are utilized to diminish defocus, coma and higher-order aberrations, as well as to decrease the slope errors caused by thermal deformation and optical polishing. The AGS is mounted on a rotatable granite platform to enable momentum-resolved RIXS measurements with scattering angles over a wide range. Several high-precision instruments developed in-house for this beamline are described briefly. The best energy resolution obtained from this AGM-AGS beamline was 12.4 meV at 530 eV, achieving a resolving power of 4.2 × 104, while the bandwidth of the incident soft X-rays was kept at 0.5 eV. To demonstrate the scientific impact of high-resolution RIXS, we present an example of momentum-resolved RIXS measurements on a high-temperature superconducting cuprate, i.e. La2-xSrxCuO4. The measurements reveal the A1g buckling phonons in superconducting cuprates, opening a new opportunity to investigate the coupling between these phonons and charge-density waves.

2.
J Synchrotron Radiat ; 28(Pt 6): 1954-1965, 2021 Nov 01.
Article in English | MEDLINE | ID: mdl-34738951

ABSTRACT

The optical design and performance of the recently opened 13A biological small-angle X-ray scattering (SAXS) beamline at the 3.0 GeV Taiwan Photon Source of the National Synchrotron Radiation Research Center are reported. The beamline is designed for studies of biological structures and kinetics in a wide range of length and time scales, from angstrom to micrometre and from microsecond to minutes. A 4 m IU24 undulator of the beamline provides high-flux X-rays in the energy range 4.0-23.0 keV. MoB4C double-multilayer and Si(111) double-crystal monochromators (DMM/DCM) are combined on the same rotating platform for a smooth rotation transition from a high-flux beam of ∼4 × 1014 photons s-1 to a high-energy-resolution beam of ΔE/E ≃ 1.5 × 10-4; both modes share a constant beam exit. With a set of Kirkpatrick-Baez (KB) mirrors, the X-ray beam is focused to the farthest SAXS detector position, 52 m from the source. A downstream four-bounce crystal collimator, comprising two sets of Si(311) double crystals arranged in a dispersive configuration, optionally collimate the DCM (vertically diffracted) beam in the horizontal direction for ultra-SAXS with a minimum scattering vector q down to 0.0004 Å-1, which allows resolving ordered d-spacing up to 1 µm. A microbeam, of 10-50 µm beam size, is tailored by a combined set of high-heat-load slits followed by micrometre-precision slits situated at the front-end 15.5 m position. The second set of KB mirrors then focus the beam to the 40 m sample position, with a demagnification ratio of ∼1.5. A detecting system comprising two in-vacuum X-ray pixel detectors is installed to perform synchronized small- and wide-angle X-ray scattering data collections. The observed beamline performance proves the feasibility of having compound features of high flux, microbeam and ultra-SAXS in one beamline.


Subject(s)
Photons , Synchrotrons , Scattering, Small Angle , Taiwan , X-Ray Diffraction , X-Rays
3.
J Eur Acad Dermatol Venereol ; 34(4): 862-872, 2020 Apr.
Article in English | MEDLINE | ID: mdl-31856347

ABSTRACT

BACKGROUND: Chancre self-healing is an important clinical feature in the early stages of syphilis infection. Wound healing may involve an important mechanism by the migration of fibroblasts filling the injured lesion. However, the specific mechanism underlying this process is still unknown. OBJECTIVES: We aimed to analyse the role of Tp0136 in the migration of fibroblasts and the related mechanism. METHODS: The migration ability of fibroblasts was detected by a wound-healing assay. RT-PCR and ELISA detected the expression of MCP-1, IL-6 and MMP-9. TLR4 expression was detected by RT-PCR. The protein levels of CCR2 and relevant signalling pathway molecules were measured by Western blotting. RESULTS: Tp0136 significantly promoted fibroblast migration. Subsequently, the levels of MCP-1 and its receptor CCR2 were increased in this process. The migration of fibroblasts was significantly inhibited by an anti-MCP-1 neutralizing antibody or CCR2 inhibitors. Furthermore, studies demonstrated that Tp0136 could activate the ERK/JNK/PI3K/NF-κB signalling pathways through TLR4 activity and that signalling pathways inhibitors could weaken MCP-1 secretion and fibroblast migration. CONCLUSIONS: These findings demonstrate that Tp0136 promotes the migration of fibroblasts by inducing MCP-1/CCR2 expression through signalling involving the TLR4, ERK, JNK, PI3K and NF-κB signalling pathways, which could contribute to the mechanism of chancre self-healing in syphilis.


Subject(s)
Chemokine CCL2/metabolism , Fibroblasts/metabolism , Receptors, CCR2/metabolism , Recombinant Proteins/metabolism , Toll-Like Receptor 4/metabolism , Treponema pallidum , Wound Healing/physiology , Blotting, Western , Cell Movement , Cells, Cultured , Enzyme-Linked Immunosorbent Assay , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction , Syphilis/pathology
4.
Neuropathol Appl Neurobiol ; 44(7): 673-686, 2018 12.
Article in English | MEDLINE | ID: mdl-29423915

ABSTRACT

AIMS: Sensory nerve degeneration and consequent abnormal sensations are the earliest and most prevalent manifestations of familial amyloid polyneuropathy (FAP) due to amyloidogenic transthyretin (TTR). FAP is a relentlessly progressive degenerative disease of the peripheral nervous system. However, there is a lack of mouse models to replicate the early neuropathic manifestations of FAP. METHODS: We established human TTR knock-in mice by replacing one allele of the mouse Ttr locus with human wild-type TTR (hTTRwt ) or human TTR with the A97S mutation (hTTRA97S ). Given the late onset of neuropathic manifestations in A97S-FAP, we investigated nerve pathology, physiology, and behavioural tests in these mice at two age points: the adult group (8 - 56 weeks) and the ageing group (> 104 weeks). RESULTS: In the adult group, nerve profiles, neurophysiology and behaviour were similar between hTTRwt and hTTRA97S mice. By contrast, ageing hTTRA97S mice showed small fibre neuropathy with decreased intraepidermal nerve fibre density and behavioural signs of mechanical allodynia. Furthermore, significant reductions in sural nerve myelinated nerve fibre density and sensory nerve action potential amplitudes in these mice indicated degeneration of large sensory fibres. The unaffected motor nerve physiology replicated the early symptoms of FAP patients, that is, sensory nerves were more vulnerable to mutant TTR than motor nerves. CONCLUSIONS: These results demonstrate that the hTTRA97S mouse model develops sensory nerve pathology and corresponding physiology mimicking A97S-FAP and provides a platform to develop new therapies for the early stage of A97S-FAP.


Subject(s)
Amyloid Neuropathies, Familial/pathology , Nerve Degeneration/pathology , Prealbumin/genetics , Sensory Receptor Cells/pathology , Amyloid Neuropathies, Familial/genetics , Animals , Disease Models, Animal , Mice , Mice, Transgenic , Nerve Degeneration/genetics
5.
Faraday Discuss ; 193: 171-185, 2016 12 12.
Article in English | MEDLINE | ID: mdl-27711809

ABSTRACT

The microscopic mechanisms of Bi electrodeposition on Au(111) and Au(100) electrodes in the overpotential regime were studied by in situ scanning tunneling microscopy with high spatial and temporal resolution. Atomic resolution images of the needle-like Bi(110) deposits formed on Au(111) reveal the central influence of covalent Bi-Bi bonds on the deposit morphology. In the straight steps along the needle edges the Bi atoms are interlinked by these bonds, whereas at the needle tip and at kinks along the needle edges dangling bonds exist, explaining the rapid structural fluctuations at these sites. For ultrathin Bi deposits on Au(100) a more open atomic arrangement was found within the surface plane, which was tentatively assigned to an epitaxially stabilised Bi(111) film. Furthermore, well-defined nanowires, consisting of zigzag chains of Bi surface atoms, were observed on this surface.

6.
Article in Zh | MEDLINE | ID: mdl-27514547

ABSTRACT

OBJECTIVE: To investigate the prevalence of hypertension in ocean seamen and major influencing factors, as well as the association between hypertension and serum microRNA21 and microRNA133a. METHODS: Health examination and a questionnaire survey were performed for 780 ocean seamen who underwent physical examination in an international travel healthcare center in Fujian, China from January to June, 2014. TaqMan RT-qPCR was used to measure the serum levels of microRNA21 and microRNA133a in seamen with hypertension. RESULTS: The prevalence of hypertension differed significantly between the ocean seamen with different ages, education levels, marital status, body mass index (BMI) values, drinking frequencies, and numbers of sailing years (P<0.05). The prevalence rate of hypertension in the ocean seamen increased with the increasing drinking frequency (χ(2)=9.02, P<0.05) , decreased with the increase in degree of education (χ(2)=11.578, P<0.05) , and increased with the increase in the number of sailing years (χ(2)=28.06, P<0.05). The hypertensive ocean seamen had significantly higher expression levels of microRNA21 and MicroRNA133a than the healthy ocean seamen (microRNA21: 7.87±5.46 vs 1.03±0.80, P<0.05; MicroRNA133a: 7.45±1.94 vs 4.52±1.15, P<0.05). The multivariate analysis showed that a high level of microRNA21 (OR=1.61, 95% CI: 1.22~2.11) , a high level of microRNA133a (OR=1.52, 95% CI: 1.24~1.87) , drinking (OR=1.64, 95% CI: 1.08~2.50) , overweight based on BMI (OR=1.18, 95%CI: 1.07~1.30) , and many sailing years (OR=2.89, 95% CI: 1.14~7.30) were risk factors for hypertension. CONCLUSION: The prevention and treatment of hypertension in ocean seamen should be enhanced. Excessive drinking should be controlled, and sailing time should be arranged reasonably. The microRNA21 and microRNA133a may be associated with the development and progression of hypertension in ocean seamen.


Subject(s)
Hypertension , Alcohol Drinking , China , Humans , MicroRNAs , Oceans and Seas , Prevalence , Risk Factors , Surveys and Questionnaires
7.
Genes Immun ; 15(6): 404-12, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24898386

ABSTRACT

Cullin 4B (CUL4B), a member of the cullin protein family, is a scaffold protein of the CUL4B-RING-E3 ligase complex that ubiquitinates intracellular proteins.CUL4B's targets include cell cycle-regulated proteins and DNA replication-related molecules. In this study, we generated myeloid-specific Cul4b-deficient mice (Cul4b(f/y);LysM-Cre(KI/KI)) to investigate the influence of Cul4b deficiency on innate immunity, especially on the function of macrophages. Our results show that an intraperitoneal injection of lipopolysaccharide (LPS) led to a significant decrease in body weights and increased leukocyte infiltrates with increased chemokines in the peritoneal cavity of Cul4b(f/y);LysM-Cre(KI/KI) mice. However, the proinflammatory cytokines, IL-6 and TNF-α did not increase in LPS-injected Cul4b(f/y);LysM-Cre(KI/KI) mice. Furthermore, bone marrow-derived macrophages from Cul4b(f/y);LysM-Cre(KI/KI) mice secreted higher levels of chemokines but lower levels of TNF-α and IL-6 upon LPS stimulation. Of note, increased proliferation of Cul4b-deficient macrophages was also observed. These results show that myeloid-specific Cul4b deficiency worsens LPS-induced peritonitis. In addition, Cul4b deficiency leads to enhanced DNA replication and proliferation, increased production of chemokines but a decreased production of proinflammatory cytokines of macrophages. Our data highlight a new role of cullin family, CUL4B, in the immune system.


Subject(s)
Cullin Proteins/immunology , Immunity, Innate/immunology , Macrophages/immunology , Peritonitis/immunology , Animals , Body Weight/immunology , Bone Marrow Cells/immunology , Bone Marrow Cells/metabolism , Cell Cycle/genetics , Cell Cycle/immunology , Cell Line, Tumor , Cell Proliferation/genetics , Chemokines/immunology , Chemokines/metabolism , Cullin Proteins/genetics , Cullin Proteins/metabolism , Gene Expression/immunology , Immunity, Innate/genetics , Immunoassay , Interleukin-6/genetics , Interleukin-6/immunology , Interleukin-6/metabolism , Leukocytes/immunology , Leukocytes/metabolism , Lipopolysaccharides , Macrophages/metabolism , Mice , Mice, Knockout , Peritonitis/chemically induced , Peritonitis/genetics , Phagocytosis/genetics , Phagocytosis/immunology , Reverse Transcriptase Polymerase Chain Reaction , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/immunology , Tumor Necrosis Factor-alpha/metabolism
8.
J Synchrotron Radiat ; 21(Pt 2): 325-32, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24562553

ABSTRACT

The design, construction and commissioning of a beamline and spectrometer for inelastic soft X-ray scattering at high resolution in a highly efficient system are presented. Based on the energy-compensation principle of grating dispersion, the design of the monochromator-spectrometer system greatly enhances the efficiency of measurement of inelastic soft X-rays scattering. Comprising two bendable gratings, the set-up effectively diminishes the defocus and coma aberrations. At commissioning, this system showed results of spin-flip, d-d and charge-transfer excitations of NiO. These results are consistent with published results but exhibit improved spectral resolution and increased efficiency of measurement. The best energy resolution of the set-up in terms of full width at half-maximum is 108 meV at an incident photon energy tuned about the Ni L3-edge.

9.
Nature ; 452(7185): 332-5, 2008 Mar 20.
Article in English | MEDLINE | ID: mdl-18354478

ABSTRACT

Equal amounts of matter and antimatter are predicted to have been produced in the Big Bang, but our observable Universe is clearly matter-dominated. One of the prerequisites for understanding this elimination of antimatter is the nonconservation of charge-parity (CP) symmetry. So far, two types of CP violation have been observed in the neutral K meson (K(0)) and B meson (B(0)) systems: CP violation involving the mixing between K(0) and its antiparticle (and likewise for B(0) and ), and direct CP violation in the decay of each meson. The observed effects for both types of CP violation are substantially larger for the B(0) meson system. However, they are still consistent with the standard model of particle physics, which has a unique source of CP violation that is known to be too small to account for the matter-dominated Universe. Here we report that the direct CP violation in charged B(+/-)-->K(+/-)pi(0) decay is different from that in the neutral B(0) counterpart. The direct CP-violating decay rate asymmetry, (that is, the difference between the number of observed B(-)-->K(-)pi(0) event versus B(+)-->K(+) pi(0) events, normalized to the sum of these events) is measured to be about +7%, with an uncertainty that is reduced by a factor of 1.7 from a previous measurement. However, the asymmetry for versus B(0)-->K(+)pi(-) is at the -10% level. Although it is susceptible to strong interaction effects that need further clarification, this large deviation in direct CP violation between charged and neutral B meson decays could be an indication of new sources of CP violation-which would help to explain the dominance of matter in the Universe.

10.
Int J Tuberc Lung Dis ; 28(2): 73-80, 2024 Feb 01.
Article in English | MEDLINE | ID: mdl-38303039

ABSTRACT

BACKGROUND: The importance of early intravenous (IV) antibiotic use for Mycobacterium abscessus complex lung diseases (MABC-LD) treatment remains unknown. METHODS: A retrospective multi-centre observational study was conducted in Taiwan. Patients who were diagnosed with and received treatment for MABC-LD from January 2007 to April 2021 were included. Treatment outcome was defined as modified microbiological cure of MABC-LD.RESULTS: Of the 89 enrolled patients, 34 (38.2%) received IV antibiotics as part of the treatment regimen. The median time to IV initiation was 1 day (IQR 1???49); 24 (70.6%) of these patients received IV agents within 4 weeks, defined as early-use. Forty-two (47.2%) patients achieved modified microbiological cure. In the multivariable logistic analysis, early IV antibiotic use was an independent factor associated with modified microbiological cure (aOR 5.32, 95% CI 1.66???17.00), whereas high radiological score (aOR 0.86, 95% CI 0.73???1.00) demonstrated negative association.CONCLUSIONS: In the present study, early use of effective IV antibiotic was prescribed in a low percentage (27%) for MABC-LD. By contrast, early IV antibiotic use was correlated with higher microbiological cure than were late or non-use. Future larger and prospective studies are needed to validate the association.


Subject(s)
Lung Diseases , Mycobacterium Infections, Nontuberculous , Mycobacterium abscessus , Humans , Anti-Bacterial Agents/therapeutic use , Mycobacterium Infections, Nontuberculous/drug therapy , Mycobacterium Infections, Nontuberculous/microbiology , Lung Diseases/drug therapy , Lung Diseases/microbiology , Prospective Studies , Retrospective Studies
11.
J Cancer Educ ; 27(3): 428-35, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22661264

ABSTRACT

The Principles and Practice of Cancer Prevention and Control course (Principles course) is offered annually by the National Cancer Institute Cancer Prevention Fellowship Program. This 4-week postgraduate course covers the spectrum of cancer prevention and control research (e.g., epidemiology, laboratory, clinical, social, and behavioral sciences) and is open to attendees from medical, academic, government, and related institutions across the world. In this report, we describe a new addition to the Principles course syllabus, which was exclusively a lecture-based format for over 20 years. In 2011, cancer prevention fellows and staff designed and implemented small group discussion sessions as part of the curriculum. The goals of these sessions were to foster an interactive environment, discuss concepts presented during the Principles course, exchange ideas, and enhance networking among the course participants and provide a teaching and leadership opportunity to current cancer prevention fellows. Overall, both the participants and facilitators who returned the evaluation forms (n=61/87 and 8/10, respectively) reported a high satisfaction with the experience for providing both an opportunity to explore course concepts in a greater detail and to network with colleagues. Participants (93%) and facilitators (100%) stated that they would like to see this component remain a part of the Principles course curriculum, and both groups provided recommendations for the 2012 program. The design, implementation, and evaluation of this initial discussion group component of the Principles course are described herein. The findings in this report will not only inform future discussion group sessions in the Principles course but may also be useful to others planning to incorporate group learning into large primarily lecture-based courses.


Subject(s)
Health Education/organization & administration , Health Status Disparities , Neoplasms/prevention & control , Consumer Behavior , Curriculum , Group Processes , Humans , Learning , Neoplasms/epidemiology , Pilot Projects , Policy , Program Evaluation
12.
Nat Med ; 4(1): 72-7, 1998 Jan.
Article in English | MEDLINE | ID: mdl-9427609

ABSTRACT

The bicyclam AMD3100 (formula weight 830) blocks HIV-1 entry and membrane fusion via the CXCR4 co-receptor, but not via CCR5. AMD3100 prevents monoclonal antibody 12G5 from binding to CXCR4, but has no effect on binding of monoclonal antibody 2D7 to CCR5. It also inhibits binding of the CXC-chemokine, SDF-1alpha, to CXCR4 and subsequent signal transduction, but does not itself cause signaling and has no effect on RANTES signaling via CCR5. Thus, AMD3100 prevents CXCR4 functioning as both a HIV-1 co-receptor and a CXC-chemokine receptor. Development of small molecule inhibitors of HIV-1 entry is feasible.


Subject(s)
Anti-HIV Agents/pharmacology , CD4-Positive T-Lymphocytes/physiology , CD4-Positive T-Lymphocytes/virology , Chemokines, CXC , HIV-1/physiology , Heterocyclic Compounds/pharmacology , Receptors, CXCR4/physiology , Antibodies, Monoclonal/pharmacology , Benzylamines , CD4 Antigens/immunology , CD4 Antigens/physiology , CD4-Positive T-Lymphocytes/drug effects , Calcium/metabolism , Carbachol/pharmacology , Cell Fusion , Cell Line , Cells, Cultured , Chemokine CCL5/pharmacology , Chemokine CXCL12 , Cyclams , Cytokines/metabolism , Cytokines/pharmacology , HIV Envelope Protein gp120/drug effects , HIV Envelope Protein gp120/metabolism , HIV-1/drug effects , Humans , Interleukin-2/pharmacology , Kinetics , Membrane Fusion/drug effects , Receptors, CCR5/physiology , Receptors, CXCR4/drug effects , Receptors, CXCR4/immunology , Signal Transduction/drug effects , Somatostatin/pharmacology
13.
Clin Microbiol Infect ; 26(11): 1555.e9-1555.e14, 2020 Nov.
Article in English | MEDLINE | ID: mdl-32061794

ABSTRACT

OBJECTIVES: Evidence of false-positive galactomannan enzyme immunoassay (GM-EIA) results associated with intravenous immunoglobulin (IVIG) administration is scarce. Here, we aimed to determine the false-positive rate of GM-EIA after IVIG administration and to identify the related factors. METHODS: Standard GM-EIA was performed using diluted and pure human IVIG samples with and without heat treatment. We also included adult patients who had at least one GM-EIA result within 1 week of IVIG administration for analysis. Those who had prior invasive aspergillosis within 1 year before IVIG therapy were excluded. The clinical characteristics and galactomannan index (GMI) kinetics between patients with false-positive and true-positive GMI were compared. RESULTS: All diluted and pure IVIG samples tested positive for GM. Heat treatment resulted in the considerable elevation of GMI. Of 48 patients with positive GM-EIA results within 1 week of IVIG administration, 22 (45.8%) were considered to have false-positive antigenaemia (false-positive group, FPG). After the completion of IVIG administration, a decline in GMI was observed in all FPG patients but in only 18 out of 26 patients (69.2%) with true-positive results (true-positive group, TPG). By 7, 14, and 18 days of IVIG administration, GMI reverted to negative values in 7/15 (46.7%), 18/20 (90%) and 22/22 (100%) FPG patients, respectively, and 6/24 (25%), 14/24 (58.3%), and 16/26 (61.5%) of TPG patients, respectively. The TPG was more likely to have two or more consecutively positive GMIs after IVIG administration than the FPG (adjusted odds ratio, 9.01; 95% confidence interval, 1.99-40.9). CONCLUSIONS: IVIG treatment may produce false-positive GM-EIA results. A positive GMI among patients receiving human IVIG should be interpreted with caution.


Subject(s)
Immunoglobulins, Intravenous/administration & dosage , Immunoglobulins, Intravenous/chemistry , Mannans/analysis , Adult , Cross-Sectional Studies , False Positive Reactions , Female , Galactose/analogs & derivatives , Hot Temperature , Humans , Immunoenzyme Techniques , Immunoglobulins, Intravenous/pharmacology , Male , Mannans/pharmacology , Mannans/therapeutic use
14.
Trends Biochem Sci ; 24(8): 300-5, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10431173

ABSTRACT

The absorption maximum of the retinal chromophore in color visual pigments is tuned by interactions with the protein (opsin) to which it is bound. Recent advances in the expression of rhodopsin-like transmembrane receptors and in spectroscopic techniques have allowed us to measure resonance Raman vibrational spectra of the retinal chromophore in recombinant visual pigments to examine the molecular basis of this spectral tuning. The dominant physical mechanism responsible for the opsin shift in color vision is the interaction of dipolar amino acid residues with the ground- and excited-state charge distributions of the chromophore.


Subject(s)
Color Perception/physiology , Retinal Pigments/chemistry , Retinal Pigments/radiation effects , Humans , In Vitro Techniques , Models, Molecular , Mutation , Protein Conformation , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/radiation effects , Retinal Pigments/genetics , Spectrum Analysis, Raman
15.
Opt Express ; 15(15): 9859-66, 2007 Jul 23.
Article in English | MEDLINE | ID: mdl-19547336

ABSTRACT

We report on an iterative design scheme for and the first experimental demonstration of active narrowband multi-wavelength filters based on aperiodically poled lithium niobate crystals. A simultaneous transmission of 8 wavelengths, each with a ~0.45-nm linewidth and nearly 100% peak transmittance, was achieved in such a device. The transmission spectrum of this device can be tuned by temperature at a rate of ~0.65 nm/ degrees C.

16.
J Pharm Biomed Anal ; 132: 109-116, 2017 Jan 05.
Article in English | MEDLINE | ID: mdl-27701037

ABSTRACT

A high performance liquid chromatography-diode array detection-tandem mass spectrometry method (HPLC-DAD-MS/MS) was developed for simultaneous determination of phenolic acids and flavonoids in djulis (Chenopodium formosanum Koidz.), a traditional Chinese herb reported to possess vital biological activities. A high yield of phenolic acids and flavonoids was attained by employing 50% ethanol in water as the extraction solvent and shaking in a 60°C water bath for 3h. A total of 8 phenolic acids and 14 flavonoids were separated and identified within 55min by using a Poroshell 120 EC-C18 column with detection at 280nm, flow rate at 0.8mL/min, column temperature at 35°C, and a gradient solvent system of 0.1% formic acid in water and acetonitrile. Two internal standards caffeic acid and kaempferol-3-O-rutinoside were used for quantitation of phenolic acids and flavonoids in djulis respectively. The amounts of phenolic acids ranged from 11.5±0.8µg/g (caffeoyl-putrescine-derivative (2)) to 1855.3±16.9µg/g (hydroxylphenylacetic acid pentoside), while the flavonoids ranged from 19.93±2.29µg/g (quercetin-3-O-(coumaryl)-rutinoside-pentoside (1)) to 257.3±2.05µg/g (rutin-O-pentoside (2)). A high recovery (89.68-97.20%) and high reproducibility was obtained for both phenolic acids and flavonoids with the relative standard deviation (RSD) for the latter ranging from 0.09-8.22% (intra-day variability) and 0.80-8.48% (inter-day variability). This method may be applied to determination of both phenolic acids and flavonoids in food products and Chinese herbs.


Subject(s)
Chenopodium/chemistry , Flavonoids/analysis , Hydroxybenzoates/analysis , Acetonitriles/chemistry , Caffeic Acids/chemistry , Chromatography, High Pressure Liquid , Flavonoids/chemistry , Hydroxybenzoates/chemistry , Kaempferols/chemistry , Limit of Detection , Plant Extracts/chemistry , Powders , Reproducibility of Results , Solvents/chemistry , Spectrometry, Mass, Electrospray Ionization , Tandem Mass Spectrometry , Temperature
17.
Nucleic Acids Res ; 29(12): 2510-6, 2001 Jun 15.
Article in English | MEDLINE | ID: mdl-11410658

ABSTRACT

The eukaryotic ribosomal 5S RNA-protein complex (5S rRNP) is formed by a co-translational event that requires 5S rRNA binding to the nascent peptide chain of eukaryotic ribosomal protein L5. Binding between 5S rRNA and the nascent chain is specific: neither the 5S rRNA nor the nascent chain of L5 protein can be substituted by other RNAs or other ribosomal proteins. The region responsible for binding 5S rRNA is located at positions 35-50 with amino acid sequence RLVIQDIKNKYNTPKYRM. Eukaryotic 5S rRNA binds a nascent chain having this sequence, but such binding is not substantive enough to form a 5S-associated RNP complex, suggesting that 5S rRNA binding to the nascent chain is amino acid sequence dependent and that formation of the 5S rRNP complex is L5 protein specific. Microinjection of 5S rRNP complex into the cytoplasm of Xenopus oocytes results in both an increase in the initial rate and also in the extent of net nuclear import of L5. This suggests that the 5S rRNP complex enhances nuclear transport of L5. We propose that 5S rRNA plays a chaperone-like role in folding of the nascent chain of L5 and directs L5 into a 5S rRNP complex for nuclear entry.


Subject(s)
Protein Biosynthesis , RNA, Ribosomal, 5S/metabolism , Ribonucleoproteins/genetics , Ribonucleoproteins/metabolism , Ribosomal Proteins/chemistry , Ribosomal Proteins/metabolism , Uracil/analogs & derivatives , Active Transport, Cell Nucleus , Amino Acid Sequence , Animals , Cross-Linking Reagents , Humans , Microinjections , Molecular Chaperones/genetics , Molecular Chaperones/metabolism , Molecular Sequence Data , Oocytes/cytology , Oocytes/metabolism , Plasmids/genetics , Polyribosomes/metabolism , Protein Binding , Protein Folding , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Ribosomal, 5S/genetics , Ribonucleoproteins/chemistry , Ribosomal Proteins/biosynthesis , Ribosomal Proteins/genetics , Sequence Deletion/genetics , Substrate Specificity , Uracil/metabolism , Xenopus laevis
18.
EBioMedicine ; 9: 257-277, 2016 Jul.
Article in English | MEDLINE | ID: mdl-27333023

ABSTRACT

Dravet syndrome is a devastating genetic brain disorder caused by heterozygous loss-of-function mutation in the voltage-gated sodium channel gene SCN1A. There are currently no treatments, but the upregulation of SCN1A healthy allele represents an appealing therapeutic strategy. In this study we identified a novel, evolutionary conserved mechanism controlling the expression of SCN1A that is mediated by an antisense non-coding RNA (SCN1ANAT). Using oligonucleotide-based compounds (AntagoNATs) targeting SCN1ANAT we were able to induce specific upregulation of SCN1A both in vitro and in vivo, in the brain of Dravet knock-in mouse model and a non-human primate. AntagoNAT-mediated upregulation of Scn1a in postnatal Dravet mice led to significant improvements in seizure phenotype and excitability of hippocampal interneurons. These results further elucidate the pathophysiology of Dravet syndrome and outline a possible new approach for the treatment of this and other genetic disorders with similar etiology.


Subject(s)
Brain/metabolism , Epilepsies, Myoclonic/pathology , NAV1.1 Voltage-Gated Sodium Channel/metabolism , RNA, Long Noncoding/metabolism , Alleles , Animals , Base Sequence , Behavior, Animal , Brain/diagnostic imaging , Cell Line , Chlorocebus aethiops , Disease Models, Animal , Electroencephalography , Epilepsies, Myoclonic/genetics , Epilepsies, Myoclonic/metabolism , Gene Expression , Gene Knock-In Techniques , Hippocampus/physiology , Humans , In Vitro Techniques , Interneurons/metabolism , Mice , Mice, Transgenic , Molecular Sequence Data , NAV1.1 Voltage-Gated Sodium Channel/chemistry , NAV1.1 Voltage-Gated Sodium Channel/genetics , Nucleic Acid Conformation , Oligonucleotides, Antisense/metabolism , Patch-Clamp Techniques , Phenotype , RNA, Long Noncoding/antagonists & inhibitors , RNA, Long Noncoding/genetics , Real-Time Polymerase Chain Reaction , Sequence Alignment , Sequence Analysis, RNA , Temperature , Up-Regulation , Vero Cells , Video Recording
19.
Gene ; 56(1): 125-35, 1987.
Article in English | MEDLINE | ID: mdl-3315856

ABSTRACT

Plasmid vectors are described that allow cloning of target DNAs at sites where they will be minimally transcribed by Escherichia coli RNA polymerase but selectively and actively transcribed by T7 RNA polymerase, in vitro or in E. coli cells. Transcription is controlled by the strong phi 10 promoter for T7 RNA polymerase, and in some cases by the T phi transcription terminator. The RNA produced can have as few as two foreign nucleotides ahead of the target sequence or can be cut by RNase III at the end of the target sequence. Target mRNAs can be translated from their own start signals or can be placed under control of start signals for the major capsid protein of T7, with the target coding sequence fused at the start codon or after the 2nd, 11th or 260th codon for the T7 protein. The controlling elements are contained on small DNA fragments that can easily be removed and used to create new expression vectors.


Subject(s)
DNA-Directed RNA Polymerases/genetics , Genetic Vectors , T-Phages/genetics , Base Sequence , DNA, Recombinant , Escherichia coli/genetics , Gene Expression Regulation , Molecular Sequence Data , Promoter Regions, Genetic , Protein Biosynthesis , T-Phages/enzymology , Terminology as Topic , Transcription, Genetic
20.
Free Radic Biol Med ; 5(4): 215-23, 1988.
Article in English | MEDLINE | ID: mdl-2908182

ABSTRACT

When exposed to oxidative stress, by oxygen radicals or H2O2, E. coli exhibited decreased growth, decreased protein synthesis, and dose-dependent increases in protein degradation. The quinone menadione induced proteolysis when cells were incubated in air, but was not effective when cells were incubated without oxygen. Anaerobically grown cells also exhibited significantly lower proteolytic capacity than did cells that were grown aerobically. Xanthine plus xanthine oxidase (which generate O2- and H2O2) caused a stimulation of proteolysis which was inhibitable by catalase, but not by superoxide dismutase: Indicating that H2O2 was responsible for the increased protein degradation. Indeed, H2O2 alone was effective in inducing increased intracellular proteolysis. Two-dimensional polyacrylamide gel electrophoresis of [3H]leucine labeled E. coli revealed greater than 50% decreases in the concentrations of 10-15 cell proteins following H2O2 or menadione exposure, while several other proteins were less severely affected. To test for the presence of soluble proteases, we prepared cell-free extracts of E. coli and incubated them with radio-labeled protein substrates. E. coli extracts degraded casein and globin polypeptides at rapid rates but showed little activity with native proteins such as superoxide dismutase, hemoglobin, bovine serum albumin, or catalase. When these same proteins were denatured by exposure to oxygen radicals or H2O2, however, they became excellent substrates for degradation in E. coli extracts. Studies with albumin revealed correlations greater than 0.95 between the degree of oxidative denaturation and proteolytic susceptibility. Pretreatment of E. coli with menadione or H2O2 did not increase the proteolytic capacity of cell extracts; indicating that neither protease activation, nor protease induction were required.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Bacterial Proteins/metabolism , Escherichia coli/metabolism , Oxygen/metabolism , Peptide Hydrolases/metabolism , Catalase/metabolism , Enzyme Activation , Escherichia coli/growth & development , Free Radicals , Glutamate-Ammonia Ligase/metabolism , Hydrogen Peroxide/metabolism , Oxidation-Reduction , Peptide Hydrolases/biosynthesis , Protein Denaturation , Superoxide Dismutase/metabolism , Vitamin K/pharmacology
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