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1.
Faraday Discuss ; 2024 May 20.
Artículo en Inglés | MEDLINE | ID: mdl-38766945

RESUMEN

Mirrors for atoms and molecules are essential tools for matter-wave optics with neutral particles. Their realization has required either a clean and atomically smooth crystal surface, sophisticated tailored electromagnetic fields, nanofabrication, or particle cooling because of the inherently short de Broglie wavelengths and strong interactions of atoms with surfaces. Here, we demonstrate reflection of He atoms from inexpensive, readily available, and robust gratings designed for light waves. Using different types of blazed gratings with different periods, we study how microscopic and macroscopic grating properties affect the mirror performance. A holographic grating with 417 nm period shows reflectivity up to 47% for He atoms, demonstrating that commercial gratings can serve as mirrors for thermal energy atoms and molecules. We also observe reflection of He2 and He3 which implies that the grating might also function as a mirror for other breakable particles that, under typical conditions, do not scatter nondestructively from a solid surface such as, e.g., metastable atoms or antihydrogen atoms.

2.
Opt Express ; 31(5): 7579-7588, 2023 Feb 27.
Artículo en Inglés | MEDLINE | ID: mdl-36859887

RESUMEN

To obtain the surface shape of an X-ray mirror with high precision, a differential deposition method was used instead of a direct removal method. To modify the mirror surface shape using the differential deposition method, it is necessary to coat it with a thick film, and the co-deposition method is used to suppress the increase in surface roughness. The addition of C to the Pt thin film, which is often used as an X-ray optical thin film, resulted in lower surface roughness compared with that with the Pt coating alone, and the stress change according to the thin film thickness was evaluated. Differential deposition controls the speed of the substrate during coating based on continuous motion. The stage was controlled by calculating the dwell time through deconvolution calculations based on the accurate measurement of the unit coating distribution and target shape. We successfully fabricated an X-ray mirror with high precision. This study indicated that an X-ray mirror surface could be manufactured by modifying the surface shape at a micrometer level through the coating. Changing the shape of existing mirrors can not only result in the manufacture of high-precision X-ray mirrors but also improve their performance.

3.
J Synchrotron Radiat ; 28(Pt 4): 1210-1215, 2021 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-34212886

RESUMEN

BL-11C, a new protein crystallography beamline, is an in-vacuum undulator-based microfocus beamline used for macromolecular crystallography at the Pohang Accelerator Laboratory and it was made available to users in June 2017. The beamline is energy tunable in the range 5.0-20 keV to support conventional single- and multi-wavelength anomalous-dispersion experiments against a wide range of heavy metals. At the standard working energy of 12.659 keV, the monochromated beam is focused to 4.1 µm (V) × 8.5 µm (H) full width at half-maximum at the sample position and the measured photon flux is 1.3 × 1012 photons s-1. The experimental station is equipped with a Pilatus3 6M detector, a micro-diffractometer (MD2S) incorporating a multi-axis goniometer, and a robotic sample exchanger (CATS) with a dewar capacity of 90 samples. This beamline is suitable for structural determination of weakly diffracting crystalline substances, such as biomaterials, including protein, nucleic acids and their complexes. In addition, serial crystallography experiments for determining crystal structures at room temperature are possible. Herein, the current beamline characteristics, technical information for users and some recent scientific highlights are described.


Asunto(s)
Cristalografía por Rayos X/instrumentación , Sustancias Macromoleculares/química , Proteínas/química , Radioisótopos de Carbono , Diseño de Equipo , Legionella/química , Muramidasa/química , Neisseria meningitidis/química , Elementos Estructurales de las Proteínas , Sincrotrones , Zymomonas/química
4.
Knee Surg Sports Traumatol Arthrosc ; 29(1): 223-229, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-32232537

RESUMEN

PURPOSE: To evaluate the incidence of intraoperative anchor pullout during arthroscopic rotator cuff repair, to compare the outcomes of different methods of managing anchor pullout, and to introduce a new technique for anchor pullout. METHODS: 1076 patients who underwent arthroscopic rotator cuff repair using a single-row repair technique were included. In 483 patients, rotator cuff repair was performed using a screw-in type anchor, and in 593 patients, soft anchors were used. When intraoperative anchor pullout occurred, it was managed by buddy screwing, anchor insertion in a different location, cement augmentation, or by bar anchoring using a threaded Steinmann pin. Plain radiography and sonography were used to check anchor locations and healing. RESULTS: Fifty-two patients experienced anchor pullout intra- or postoperatively (48 and four patients, respectively). Anchor pullouts were more frequently observed for larger tears, women, older patients, and in patients with preoperative stiffness (limitations of both active and passive movements of the affected shoulder joint). For screw-in type anchors, pullout during surgery occurred in 16 patients (3.3%, 16/483), and all were managed using the buddy screwing technique. For soft anchor cases, pullout occurred in 32 patients (5.4%, 32/593) and was managed by anchor insertion in a different location (17 patients), cement augmentation (two patients), or bar anchoring using a threaded Steinmann pin (13 patients). Three patients managed by buddy screwing and two patients managed by anchor insertion in a different location had anchor failure after repair. Tendon healing at 6 months was observed in 12/16 patients treated by buddy screwing, 11/17 treated by anchor insertion in a different location, 2/2 treated by cement augmentation, and 12/13 treated by bar anchoring with a threaded Steinmann pin. CONCLUSION: Intraoperative anchor pullout during arthroscopic rotator cuff repair is an uncommon but cumbersome complication. There are some techniques already introduced to deal with this complication. In comparison, not one technique is overwhelmingly superior to others; however, our new technique which is bar anchoring with a threaded Steinmann pin could be another solution, since it could utilize primary anchor sites and results appear to be acceptable. Level of evidence III.


Asunto(s)
Artroscopía/métodos , Lesiones del Manguito de los Rotadores/cirugía , Anclas para Sutura , Anciano , Artroscopía/efectos adversos , Artroscopía/instrumentación , Cementos para Huesos , Femenino , Humanos , Complicaciones Intraoperatorias/cirugía , Masculino , Persona de Mediana Edad , Radiografía , Manguito de los Rotadores/diagnóstico por imagen , Manguito de los Rotadores/cirugía , Lesiones del Manguito de los Rotadores/diagnóstico por imagen , Técnicas de Sutura , Ultrasonografía
5.
J Synchrotron Radiat ; 26(Pt 4): 1101-1109, 2019 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-31274433

RESUMEN

A wake monochromator based on a large-area diamond single crystal for hard X-ray self-seeding has been successfully installed and commissioned in the hard X-ray free-electron laser (FEL) at the Pohang Accelerator Laboratory with international collaboration. For this commissioning, the self-seeding was demonstrated with a low bunch charge (40 pC) and the nominal bunch charge (180 pC) of self-amplified spontaneous emission (SASE) operation. The FEL pulse lengths were estimated as 7 fs and 29.5 fs, respectively. In both cases, the average spectral brightness increased by more than three times compared with the SASE mode. The self-seeding experiment was demonstrated for the first time using a crystal with a thickness of 30 µm, and a narrow bandwidth of 0.22 eV (full width at half-maximum) was obtained at 8.3 keV, which confirmed the functionality of a crystal with such a small thickness. In the nominal bunch-charge self-seeding experiment, the histogram of the intensity integrated over a 1 eV bandwidth showed a well defined Gaussian profile, which is evidence of the saturated FEL and a minimal electron-energy jitter (∼1.2 × 10-4) effect. The corresponding low photon-energy jitter (∼2.4 × 10-4) of the SASE FEL pulse, which is two times lower than the Pierce parameter, enabled the seeding power to be maximized by maintaining the spectral overlap between SASE FEL gain and the monochromator.

6.
J Synchrotron Radiat ; 25(Pt 1): 289-292, 2018 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-29271778

RESUMEN

The Pohang Accelerator Laboratory X-ray Free-Electron Laser (PAL-XFEL) is a recently commissioned X-ray free-electron laser (XFEL) facility that provides intense ultrashort X-ray pulses based on the self-amplified spontaneous emission process. The nano-crystallography and coherent imaging (NCI) hutch with forward-scattering geometry is located at the hard X-ray beamline of the PAL-XFEL and provides opportunities to perform serial femtosecond crystallography and coherent X-ray diffraction imaging. To produce intense high-density XFEL pulses at the interaction positions between the X-rays and various samples, a microfocusing Kirkpatrick-Baez (KB) mirror system that includes an ultra-precision manipulator has been developed. In this paper, the design of a KB mirror system that focuses the hard XFEL beam onto a fixed sample point of the NCI hutch, which is positioned along the hard XFEL beamline, is described. The focusing system produces a two-dimensional focusing beam at approximately 2 µm scale across the 2-11 keV photon energy range. XFEL pulses of 9.7 keV energy were successfully focused onto an area of size 1.94 µm × 2.08 µm FWHM.

7.
J Shoulder Elbow Surg ; 25(3): 478-86, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26541205

RESUMEN

BACKGROUND: Medial row failure has been reported in the suture bridge technique of rotator cuff repair. This study compared the healing response of suture bridge configuration repair (SBCR) and parallel type transosseous repair (PTR). METHODS: Acute rotator cuff repair was performed in 32 rabbits. Both shoulders were repaired using PTR or SBCR. In PTR, simple PTR was performed through 2 parallel transosseous tunnels created using a microdrill. In SBCR, 2 additional crisscross transosseous tunnels were added to mimic arthroscopic SBCR. At 1, 2, and 5 weeks postoperatively, comparative biomechanical testing was performed in 8 rabbits, and histologic analysis, including immunohistochemical staining for CD31, was performed in 4 rabbits. RESULTS: Failure loads at 1 week (38.12 ± 20.43 N vs 52.00 ± 27.23 N; P = .284) and 5 weeks (97.93 ± 48.35 N vs 119.60 ± 60.81 N; P = .218) were not statistically different between the SBCR and PTR groups, respectively, but were significantly lower in the SBCR group than in the PTR group (23.56 ± 13.56 N vs. 44.25 ± 12.53 N; P = .009), respectively, at 2 weeks. Markedly greater fibrinoid deposition was observed in the SBCR group than in the PTR group at 2 weeks. For vascularization, there was a tendency that more vessels could be observed in PTR than in SBCR at 2 weeks (15.9 vs 5.6, P = .068). CONCLUSIONS: In a rabbit acute rotator cuff repair model, SBCR exhibited inferior mechanical strength, and fewer blood vessels were observed at the healing site at 2 weeks postoperatively. Medial row tendon failure was more common in SBCR. Surgeons should consider the clinical effect of SBCR when performing rotator cuff repair.


Asunto(s)
Procedimientos Ortopédicos/métodos , Manguito de los Rotadores/cirugía , Articulación del Hombro/fisiopatología , Técnicas de Sutura , Traumatismos de los Tendones/cirugía , Cicatrización de Heridas , Animales , Fenómenos Biomecánicos , Masculino , Neovascularización Fisiológica , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/análisis , Conejos , Manguito de los Rotadores/irrigación sanguínea , Lesiones del Manguito de los Rotadores , Rotura/cirugía , Técnicas de Sutura/efectos adversos
8.
Opt Express ; 23(22): 29032-7, 2015 Nov 02.
Artículo en Inglés | MEDLINE | ID: mdl-26561172

RESUMEN

We evaluated the irradiation damage induced by hard X-ray free-electron lasers to platinum/carbon multilayers intended for use in a focusing reflective mirror. In order to determine the damage threshold, we compared X-ray reflectivities before and after irradiation at the first-order Bragg angle using a focused X-ray free-electron laser with a beam size of approximately 1 µm and a pulse energy ranging from 0.01 to 10 µJ at a photon energy of 10 keV. We confirmed that the damage threshold of the platinum/carbon multilayer with a bilayer period of 3 nm was 0.051 µJ/µm(2), which is sufficiently higher than that in practical applications.

9.
Pain Med ; 15(12): 2037-45, 2014 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-25288391

RESUMEN

OBJECTIVES: To investigate the role of preoperative pain sensitivity and preoperative symptom severity for prediction of postoperative pain intensity after lumbar spine surgery. METHODS: This study consisted of two groups who underwent decompression surgery alone (62 patients) or decompression with fusion surgery (37 patients) for lumbar spinal stenosis (LSS). Pain Sensitivity Questionnaire (PSQ) and visual analog pain scale (VAS) for back pain and leg pain were collected preoperatively with detailed medical history. The assessment was performed immediately after surgery when the patients had completely recovered and regained their complete consciousness from general anesthesia (H0) and subsequently 4, 8, 18, 30, 48, and 72 hours (H4, H8, H18, H30, H48, and H72) thereafter as they recovered. RESULTS: Both groups showed a decrease in back pain and leg pain with the time postoperatively. In fusion group, preoperative VAS for back pain was significantly correlated with postoperative VAS for back pain at H0, H4, H8, and H18, and PSQ minor/total PSQ also showed a significant correlation with postoperative back pain at H48 and H72. In contrast, only total PSQ and PSQ minors were significantly correlated with postoperative back pain at H18 and H30 in decompression group. Hierarchical regression analysis finally showed that each preoperative back pain and PSQ minor was predictive of immediate postoperative back pain (from H0 to H18) in fusion group and delayed postoperative back pain (H18, H30) in decompression group. CONCLUSIONS: The study highlights that each preoperative back pain and individual pain sensitivity could predict the different aspects of postoperative pain after lumbar surgery.


Asunto(s)
Dolor de Espalda/fisiopatología , Hiperalgesia/complicaciones , Umbral del Dolor , Dolor Postoperatorio/etiología , Estenosis Espinal/cirugía , Anciano , Descompresión Quirúrgica/efectos adversos , Femenino , Humanos , Vértebras Lumbares , Masculino , Persona de Mediana Edad , Dimensión del Dolor , Periodo Preoperatorio
10.
Opt Express ; 21(13): 15382-8, 2013 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-23842324

RESUMEN

We evaluated the ablation thresholds of optical materials by using hard X-ray free electron laser. A 1-µm-focused beam with 10-keV of photon energy from SPring-8 Angstrom Compact free electron LAser (SACLA) was irradiated onto silicon and SiO2 substrates, as well as the platinum and rhodium thin films on these substrates, which are widely used for optical materials such as X-ray mirrors. We designed and installed a dedicated experimental chamber for the irradiation experiments. For the silicon substrate irradiated at a high fluence, we observed strong mechanical cracking at the surface and a deep ablation hole with a straight side wall. We confirmed that the ablation thresholds of uncoated silicon and SiO2 substrates agree with the melting doses of these materials, while those of the substrates under the metal coating layer are significantly reduced. The ablation thresholds obtained here are useful criteria in designing optics for hard X-ray free electron lasers.

11.
Clin Shoulder Elb ; 25(2): 121-128, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-35698781

RESUMEN

BACKGROUND: To determine the reasons and factors that contribute to the cancellations of shoulder surgeries at a tertiary referral center and to analyze the characteristics of these patients. METHODS: Patients scheduled for shoulder surgery from June 2017 to July 2019 were allocated to a surgery group (n=224) or a cancellation group (n=96). These groups were compared with respect to patient characteristics, types of surgery, distance from patient's home to the hospital, traveling time to the hospital, and waiting period before surgery. Reasons for cancellation and responses were acquired using a telephone interview and were subsequently analyzed. RESULTS: The cancellation group was older, had a less frequent history of trauma, and had a lower proportion of patients undergoing arthroscopic rotator cuff repair than the surgery group (p=0.009, p=0.014, and p=0.017, respectively). In addition, mean distance from the patients' homes to the hospital and preoperative waiting time were both longer in the cancellation group (p=0.001 and p<0.01, respectively). The most common reason given for cancellation was another medical condition (28.1%). CONCLUSIONS: Older age, need for arthroscopic rotator cuff repair surgery, longer distance from the patient's home to the hospital, and longer waiting period significantly increased the chance of cancellation. The main reason for canceling surgery was a concurrent medical condition. Therefore, identification of other medical conditions in advance is an important consideration when surgeons recommend shoulder surgery to patients. Surgeons should also consider patient's age, type of surgery, distance from the hospital, and waiting time when assessing the possibility of surgery cancellation.

12.
Sci Rep ; 11(1): 21787, 2021 11 08.
Artículo en Inglés | MEDLINE | ID: mdl-34750381

RESUMEN

Photosystem I (PS I) has a symmetric structure with two highly similar branches of pigments at the center that are involved in electron transfer, but shows very different efficiency along the two branches. We have determined the structure of cyanobacterial PS I at room temperature (RT) using femtosecond X-ray pulses from an X-ray free electron laser (XFEL) that shows a clear expansion of the entire protein complex in the direction of the membrane plane, when compared to previous cryogenic structures. This trend was observed by complementary datasets taken at multiple XFEL beamlines. In the RT structure of PS I, we also observe conformational differences between the two branches in the reaction center around the secondary electron acceptors A1A and A1B. The π-stacked Phe residues are rotated with a more parallel orientation in the A-branch and an almost perpendicular confirmation in the B-branch, and the symmetry breaking PsaB-Trp673 is tilted and further away from A1A. These changes increase the asymmetry between the branches and may provide insights into the preferential directionality of electron transfer.


Asunto(s)
Complejo de Proteína del Fotosistema I/química , Vitamina K 1/química , Cristalografía por Rayos X , Fotosíntesis , Estructura Terciaria de Proteína , Temperatura , Thermosynechococcus
13.
Langmuir ; 26(14): 11928-33, 2010 Jul 20.
Artículo en Inglés | MEDLINE | ID: mdl-20557075

RESUMEN

To obtain evenly distributed pure Ag particles with a narrow size distribution on a polymer membrane, a novel activation procedure with an environmentally friendly, cost-effective method was utilized as a pretreatment before electroless Ag deposition. The pretreatment was first performed on an untreated membrane surface by collecting ultrafine ambient spark-generated Ag aerosol particles. After annealing, the electroless Ag film was fabricated on the collected aerosol particles in the Ag electroless bath. Experimental characterizations showed that the ultrafine Ag particles were uniformly anchored onto the membrane surface through pretreatment, resulting in a pure Ag film of closely packed particles with a narrow size distribution on the membrane, and the properties were comparable to those of an Ag film on wet Sn-Ag-activated membranes.


Asunto(s)
Tecnología Química Verde/métodos , Plata/química , Aerosoles , Microscopía Electrónica de Rastreo , Estaño/química
14.
Clin Shoulder Elb ; 23(1): 48-58, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33330234

RESUMEN

Although the prevalence of rotator cuff tears is dependent on the size, 11% to 94% of patients experience retear or healing failure after rotator cuff repair. Treatment of patients with failed rotator cuff repair ranges widely, from conservative treatment to arthroplasty. This review article attempts to summarize the most recent and relevant surgical options for failed rotator cuff repair patients, and the outcomes of each treatment, except arthroplasty.

15.
Am J Sports Med ; 48(11): 2669-2676, 2020 09.
Artículo en Inglés | MEDLINE | ID: mdl-32795189

RESUMEN

BACKGROUND: Postoperative stiffness after rotator cuff repair is a common complication that can lead to poor outcomes and patient discomfort. The application of an antiadhesive agent at the time of repair recently became an option for clinicians, but little information is available on its effects. PURPOSE: To evaluate and compare retear rates, the incidence of postoperative stiffness, and the clinical outcomes of patients who underwent cuff repair with or without the application of an antiadhesive agent. STUDY DESIGN: Cohort study; Level of evidence, 3. METHODS: Among 296 patients who underwent arthroscopic rotator cuff repair surgery, we compared the outcomes of those injected with a thermosensitive gel antiadhesive agent into the subacromial space (112 cases) versus noninjected controls (184 cases). Retear rates in the 2 groups were determined by magnetic resonance imaging at 1 year after surgery. Shoulder joint range of motion and functional scores were evaluated serially. RESULTS: The rate of retear was significantly lower in the injection group (20/112 cases; 17.9%) than the control group (53/184 cases; 28.8%) (P = .034). Postoperative stiffness was not significantly different between the 2 groups (P = .710). Among the data regarding range of motion, only forward flexion at 6 months after surgery showed superior results in the injection group. Functional scores showed conflicting results: The control group had better visual analog scale scores for pain (injection vs control: 2.17 vs 1.68 at 6 months; 1.82 vs 1.28 at 12 months), American Shoulder and Elbow Surgeons scores (79.89 vs 89.64 at 12 months), and simple shoulder test scores (8.70 vs 10.06 at 12 months), whereas the injection group had better Constant-Murley scores (injection vs control: 59.49 vs 55.60 at 3 months; 77.35 vs 71.98 at 6 months; 87.28 vs 81.56 at 12 months). CONCLUSION: The tendon healing rate was significantly higher in the group receiving an antiadhesive agent than in the control group. No intergroup difference was seen in the occurrence of postoperative stiffness. However, the pain-related functional score showed inferior results in the injection group at 12 months. The biological action of antiadhesive agents in rotator cuff repair should be further evaluated.


Asunto(s)
Lesiones del Manguito de los Rotadores , Manguito de los Rotadores , Adherencias Tisulares , Viscosuplementos , Artroscopía , Estudios de Cohortes , Humanos , Imagen por Resonancia Magnética , Rango del Movimiento Articular , Lesiones del Manguito de los Rotadores/cirugía , Adherencias Tisulares/prevención & control , Resultado del Tratamiento , Viscosuplementos/uso terapéutico
16.
Clin Orthop Surg ; 12(3): 371-378, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32904028

RESUMEN

BACKGROUD: Lesions of the long head of the biceps tendon (LHBT) are one of the most common pathologies in patients with a rotator cuff tear. Although various procedures have been shown to be effective for treating LHBT lesions during rotator cuff repair, no consensus has been reached regarding the most effective treatment. The purpose of this study was to compare the outcomes of tenotomy vs subpectoral tenodesis of the LHBT in arthroscopic rotator cuff repair. METHODS: The records of 135 patients who underwent arthroscopic rotator cuff repair with biceps tenotomy or subpectoral tenodesis for a partial LHBT tear of > 50% were initially reviewed. Finally, 77 patients (38 patients with tenotomy and 39 patients with subpectoral tenodesis) with an intact rotator cuff, who underwent a functional evaluation at 1 year postoperatively, were enrolled in this retrospective study. RESULTS: The average follow-up was 13.3 ± 4.36 months (13.2 ± 1.4 months in the tenotomy group and 13.6 ± 2.7 months in the subpectoral tenodesis group; p = 0.416). Demographic and surgical data were not significantly different between the 2 groups. Preoperatively, biceps groove tenderness, Speed's test, and Yergason test results were positive in 27.3%, 27.3%, and 10.4% of the study subjects, respectively. Compared with preoperative values, all functional scores including shoulder muscle power were significantly improved postoperatively, and no significant intergroup difference was observed (all p > 0.05). A visible Popeye deformity was not encountered in either group at the final follow-up. Eight patients in the tenotomy group and 7 patients in the subpectoral tenodesis group complained of mild anterior shoulder pain (p = 0.731), and 4 patients in each group complained of groove tenderness (p = 0.969). No surgical or postoperative complication occurred in either group. CONCLUSIONS: Both biceps tenotomy and subpectoral tenodesis performed during rotator cuff repair improved pain and function and resulted in comparable clinical outcomes. Residual symptoms associated with the remnant LHBT in the groove may not be a problem after adhesion of LHBT.


Asunto(s)
Artroscopía/métodos , Lesiones del Manguito de los Rotadores/cirugía , Traumatismos de los Tendones/cirugía , Tenodesis/métodos , Tenotomía/métodos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Dimensión del Dolor , Estudios Retrospectivos
17.
J Appl Crystallogr ; 53(Pt 2): 477-485, 2020 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-32280322

RESUMEN

Microcrystal delivery methods are pivotal in the use of serial femtosecond crystallography (SFX) to resolve the macromolecular structures of proteins. Here, the development of a novel technique and instruments for efficiently delivering microcrystals for SFX are presented. The new method, which relies on a one-dimensional fixed-target system that includes a microcrystal container, consumes an extremely low amount of sample compared with conventional two-dimensional fixed-target techniques at ambient temperature. This novel system can deliver soluble microcrystals without highly viscous carrier media and, moreover, can be used as a microcrystal growth device for SFX. Diffraction data collection utilizing this advanced technique along with a real-time visual servo scan system has been successfully demonstrated for the structure determination of proteinase K microcrystals at 1.85 Šresolution.

18.
J Neurochem ; 111(5): 1202-12, 2009 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-19780901

RESUMEN

Midbrain dopamine (mDA) neurons play critical roles in the regulation of voluntary movement and their dysfunction is associated with Parkinson's disease. Pitx3 has been implicated in the proper development of mDA neurons in the substantia nigra pars compacta, which are selectively lost in Parkinson's disease. However, the basic mechanisms underlying its role in mDA neuron development and/or survival are poorly understood. Toward this goal, we sought to identify downstream target genes of Pitx3 by comparing gene expression profiles in mDA neurons of wild-type and Pitx3-deficient aphakia mice. This global gene expression analysis revealed many potential target genes of Pitx3; in particular, the expression of vesicular monoamine transporter 2 and dopamine transporter, responsible for dopamine storage and reuptake, respectively, is greatly reduced in mDA neurons by Pitx3 ablation. In addition, gain-of-function analyses and chromatin immunoprecipitation strongly indicate that Pitx3 may directly activate transcription of vesicular monoamine transporter 2 and dopamine transporter genes, critically contributing to neurotransmission and/or survival of mDA neurons. As the two genes have been known to be regulated by Nurr1, another key dopaminergic transcription factor, we propose that Pitx3 and Nurr1 may coordinately regulate mDA specification and survival, at least in part, through a merging and overlapping downstream pathway.


Asunto(s)
Proteínas de Transporte de Dopamina a través de la Membrana Plasmática/metabolismo , Dopamina/metabolismo , Proteínas de Homeodominio/metabolismo , Mesencéfalo/citología , Neuronas/metabolismo , Factores de Transcripción/metabolismo , Proteínas de Transporte Vesicular de Monoaminas/metabolismo , Animales , Línea Celular Transformada , Inmunoprecipitación de Cromatina/métodos , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática/genética , Embrión de Mamíferos , Perfilación de la Expresión Génica/métodos , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Regulación del Desarrollo de la Expresión Génica/genética , Regulación del Desarrollo de la Expresión Génica/fisiología , Proteínas de Homeodominio/genética , Humanos , Factor Inhibidor de Leucemia/farmacología , Ratones , Ratones Noqueados , Microdisección/métodos , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Factores de Transcripción/deficiencia , Factores de Transcripción/genética , Proteínas de Transporte Vesicular de Monoaminas/genética
19.
Sci Rep ; 9(1): 2525, 2019 02 21.
Artículo en Inglés | MEDLINE | ID: mdl-30792457

RESUMEN

Serial femtosecond crystallography (SFX) provides opportunities to observe the dynamics of macromolecules without causing radiation damage at room temperature. Although SFX provides a biologically more reliable crystal structure than provided by the existing synchrotron sources, there are limitations due to the consumption of many crystal samples. A viscous medium as a carrier matrix reduces the flow rate of the crystal sample from the injector, thereby dramatically reducing sample consumption. However, the currently available media cannot be applied to specific crystal samples owing to reactions between the viscous medium and crystal sample. The discovery and characterisation of a new delivery medium for SFX can further expand its use. Herein, we report the preparation of a polyacrylamide (PAM) injection matrix to determine the crystal structure with an X-ray free-electron laser. We obtained 11,936 and 22,213 indexed images using 0.5 mg lysozyme and 1.0 mg thermolysin, respectively. We determined the crystal structures of lysozyme and thermolysin delivered in PAM at 1.7 Å and 1.8 Å resolutions. The maximum background scattering from PAM was lower than monoolein, a commonly used viscous medium. Our results show that PAM can be used as a sample delivery media in SFX studies.

20.
Sci Rep ; 9(1): 6971, 2019 05 06.
Artículo en Inglés | MEDLINE | ID: mdl-31061502

RESUMEN

Fixed-target serial femtosecond crystallography (FT-SFX) was an important advance in crystallography by dramatically reducing sample consumption, while maintaining the benefits of SFX for obtaining crystal structures at room temperature without radiation damage. Despite a number of advantages, preparation of a sample holder for the sample delivery in FT-SFX with the use of many crystals in a single mount at ambient temperature is challenging as it can be complicated and costly, and thus, development of an efficient sample holder is essential. In this study, we introduced a nylon mesh-based sample holder enclosed by a polyimide film. This sample holder can be rapidly manufactured using a commercially available nylon mesh with pores of a desired size at a low cost without challenging technology. Furthermore, this simple device is highly efficient in data acquisition. We performed FT-SFX using a nylon mesh-based sample holder and collected over 130,000 images on a single sample holder using a 30 Hz X-ray pulse for 1.2 h. We determined the crystal structures of lysozyme and glucose isomerase using the nylon mesh at 1.65 and 1.75 Å, respectively. The nylon mesh exposed to X-rays produced very low levels of background scattering at 3.75 and 4.30 Å, which are negligible for data analysis. Our method provides a simple and rapid but highly efficient way to deliver samples for FT-SFX.

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