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Brefeldin A (BFA) disrupts the structure of the Golgi apparatus to trigger ER stress signaling pathways. On the other hand, treatment with BFA induces the activation of CREB3, the protein structure of which is similar to that of ATF6. In this study, we established Neuro2a cells in which three different transcription factors, namely, ATF4, ATF3 and CREB3, were deficient using the CRISPR/Cas9 approach, and we investigated the BFA-induced ER and Golgi stress response in these cells. BFA treatment rapidly induced ATF4, ATF3, Herp and GADD153 protein expression in Neuro2a cells. ATF4-deficient Neuro2a cells exhibited significantly decreased mRNA and protein expression of ATF3 and Herp but not GADD153; however, cells deficient in ATF3 exhibited minimal effects on GADD34, GADD153 and Herp expression. The cleavage of CREB3 in Neuro2a cells was triggered by BFA; however, the expression of several ER and Golgi stress-related factors was hardly influenced by the CREB3 deficiency in these Neuro2a cells. This study shows that CREB3 minimally associates with typical ER stress-inducible responses in Neuro2a cells. Therefore, identification and characterization of the downstream transcriptional targets of CREB3 is required to clarify not only Golgi stress response but also its relationship with ER stress signaling pathways.
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Brefeldina A/farmacologia , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Retículo Endoplasmático/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Complexo de Golgi/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Linhagem Celular , Retículo Endoplasmático/genética , Estresse do Retículo Endoplasmático/genética , Complexo de Golgi/genética , Camundongos , Transdução de Sinais/genéticaRESUMO
We performed expression and functional analysis of mouse CREB3 regulatory factor (CREBRF) in Neuro2a cells by constructing several expression vectors. Overexpressed full-length (FL) CREBRF protein was stabilized by MG132; however, the intrinsic CREBRF expression in Neuro2a cells was negligible under all conditions. On the other hand, N- or C-terminal deletion of CREBRF influenced its stability. Cotransfection of CREBRF together with GAL4-tagged FL CREB3 increased luciferase reporter activity, and only the N-terminal region of CREBRF was sufficient to potentiate luciferase activity. Furthermore, this positive effect of CREBRF was also observed in cells expressing GAL4-tagged cleaved CREB3, although CREBRF hardly influenced the protein stability of NanoLuc-tagged cleaved CREB3 or intracellular localization of EGFP-tagged one. In conclusion, this study suggests that CREBRF, a quite unstable proteasome substrate, positively regulates the CREB3 pathway, which is distinct from the canonical ER stress pathway in Neuro2a cells.
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Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Animais , Linhagem Celular Tumoral , Células Cultivadas , Expressão Gênica , Genes Reporter , Camundongos , Plasmídeos/genética , TransfecçãoRESUMO
While some MRI systems offer a "pause" function, combining it with the PROPELLER method for image quality improvement remains underexplored. This study investigated whether repositioning the head after pausing during PROPELLER imaging enhances image quality. All brain phantom images in this study were obtained using a 3.0 T MRI and acquired using the fast spin-echo T2WI-based PROPELLER with motion correction. By combining the angle of rotational motion of the head phantom and the number of repositioning after a pause, two studies including seven trials were performed. Increasing the rotation angle decreased the image quality; however, pausing the image and repositioning the head phantom to the original angle improved the image quality. A similar result was obtained by repositioning the angle closer to its original angle. Experiments with multiple head movements showed that pausing the scan and repositioning the phantom with each movement improved image quality.
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Encéfalo , Processamento de Imagem Assistida por Computador , Imageamento por Ressonância Magnética , Imagens de Fantasmas , Imageamento por Ressonância Magnética/instrumentação , Imageamento por Ressonância Magnética/métodos , Encéfalo/diagnóstico por imagem , Encéfalo/fisiologia , Processamento de Imagem Assistida por Computador/métodos , Humanos , MovimentoRESUMO
This is an explanatory paper on Sun Il Kwon et al., Nat. Photon. 15: 914-918, 2021 and some parts of this manuscript are translated from the paper. Medical imaging modalities such as X-ray computed tomography, Magnetic resonance imaging, positron emission tomography (PET), and single photon emission computed tomography, require image reconstruction processes, consequently constraining them to form cylindrical shapes. However, among them, only PET can use additional information, so called time of flight, on an event-by-event basis. If coincidence time resolution (CTR) of PET detectors improved to 30 ps, which corresponds to spatial resolution of 4.5 mm, directly localizing electron-positron annihilation point is possible, allowing us to circumvent image reconstruction processes and free us from the geometric constraint. We call this concept direct positron emission imaging (dPEI). We have developed ultrafast radiation detectors by focusing on Cherenkov photon detection. Furthermore, the CTR of 32 ps being equivalent to 4.8 mm spatial resolution is achieved by combining deep learning-based signal processing with the detectors. In this article, we explain how we developed the detectors and demonstrated the first dPEI using different types of phantoms, how we will tackle limitations to be addressed to make the dPEI more practical, and how dPEI will emerge as an imaging modality in nuclear medicine.
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Processamento de Imagem Assistida por Computador , Tomografia por Emissão de Pósitrons , Processamento de Imagem Assistida por Computador/métodos , Imagens de Fantasmas , Fótons , Tomografia por Emissão de Pósitrons/instrumentação , Tomografia por Emissão de Pósitrons/métodos , Fatores de TempoRESUMO
Erythrocyte invasion by merozoites is an obligatory stage of Plasmodium infection and is essential to disease progression. Proteins in the apical organelles of merozoites mediate the invasion of erythrocytes and are potential malaria vaccine candidates. Rhoptry-associated, leucine zipper-like protein 1 (RALP1) of Plasmodium falciparum was previously found to be specifically expressed in schizont stages and localized to the rhoptries of merozoites by immunofluorescence assay (IFA). Also, RALP1 has been refractory to gene knockout attempts, suggesting that it is essential for blood-stage parasite survival. These characteristics suggest that RALP1 can be a potential blood-stage vaccine candidate antigen, and here we assessed its potential in this regard. Antibodies were raised against recombinant RALP1 proteins synthesized by using the wheat germ cell-free system. Immunoelectron microscopy demonstrated for the first time that RALP1 is a rhoptry neck protein of merozoites. Moreover, our IFA data showed that RALP1 translocates from the rhoptry neck to the moving junction during merozoite invasion. Growth and invasion inhibition assays revealed that anti-RALP1 antibodies inhibit the invasion of erythrocytes by merozoites. The findings that RALP1 possesses an erythrocyte-binding epitope in the C-terminal region and that anti-RALP1 antibodies disrupt tight-junction formation, are evidence that RALP1 plays an important role during merozoite invasion of erythrocytes. In addition, human sera collected from areas in Thailand and Mali where malaria is endemic recognized this protein. Overall, our findings indicate that RALP1 is a rhoptry neck erythrocyte-binding protein and that it qualifies as a potential blood-stage vaccine candidate.
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Antígenos de Protozoários/metabolismo , Eritrócitos/parasitologia , Vacinas Antimaláricas/imunologia , Plasmodium falciparum/fisiologia , Proteínas de Protozoários/metabolismo , Receptores de Superfície Celular/metabolismo , Adulto , Animais , Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/imunologia , Imunofluorescência , Humanos , Vacinas Antimaláricas/genética , Mali , Microscopia Imunoeletrônica , Proteínas de Protozoários/imunologia , Receptores de Superfície Celular/imunologia , Soro/imunologia , TailândiaRESUMO
BACKGROUND: Intensity-modulated radiation therapy (IMRT) requires delivery quality assurance (DQA) to ensure treatment accuracy and safety. Irradiation techniques such as helical tomotherapy (HT) have become increasingly complex, rendering conventional verification methods insufficient. This study aims to develop a novel DQA system to simultaneously verify dose distribution and multi-leaf collimator (MLC) opening during HT. METHODS: We developed a prototype detector consisting of a cylindrical plastic scintillator (PS) and a cooled charge-coupled device (CCD) camera. Scintillation light was recorded using a CCD camera. A TomoHDA (Accuray Inc.) was used as the irradiation device. The characteristics of the developed system were evaluated based on the light intensity. The IMRT plan was irradiated onto the PS to record a moving image of the scintillation light. MLC opening and light distribution were obtained from the recorded images. To detect MLC opening, we placed a region of interest (ROI) on the image, corresponding to the leaf position, and analyzed the temporal change in the light intensity within each ROI. Corrections were made for light changes due to differences in the PS shape and irradiation position. The corrected light intensity was converted into the leaf opening time (LOT), and an MLC sinogram was constructed. The reconstructed MLC sinogram was compared with that calculated using the treatment planning system (TPS). Light distribution was obtained by integrating all frames obtained during IMRT irradiation. The light distribution was compared with the dose distribution calculated using the TPS. RESULTS: The LOT and the light intensity followed a linear relationship. Owing to MLC movements, the sensitivity and specificity of the reconstructed sinogram exceeded 97%, with an LOT error of - 3.9 ± 7.8%. The light distribution pattern closely resembled that of the dose distribution. The average dose difference and the pass rate of gamma analysis with 3%/3 mm were 1.4 ± 0.2% and 99%, respectively. CONCLUSION: We developed a DQA system for simultaneous and accurate verification of both dose distribution and MLC opening during HT.
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Radioterapia de Intensidade Modulada , Humanos , Radioterapia de Intensidade Modulada/métodos , Dosagem Radioterapêutica , Planejamento da Radioterapia Assistida por Computador/métodos , Imagens de FantasmasRESUMO
Since the early 2000s, many types of positron emission tomography (PET) scanners dedicated to breast imaging for the diagnosis of breast cancer have been introduced. However, conventional performance evaluation methods developed for whole-body PET scanners cannot be used for such devices. In this study, we developed phantom tools for evaluating the quantitative accuracy of positron emission mammography (PEM) and dedicated-breast PET (dbPET) scanners using novel traceable point-like 68Ge/68 Ga sources. The PEM phantom consisted of an acrylic cube (100 × 100 × 40 mm) and three point-like sources. The dbPET phantom comprised an acrylic cylinder (ø100 × 100 mm) and five point-like sources. These phantoms were used for evaluating the fundamental responses of clinical PEM and dbPET scanners to point-like inputs in a medium. The results showed that reasonable recovery values were obtained based on region-of-interest analyses of the reconstructed images. The developed phantoms using traceable 68Ge/68 Ga point-like sources were useful for evaluating the physical characteristics of PEM and dbPET scanners. Thus, they offer a practical, reliable, and universal measurement scheme for evaluating various types of PET scanners using common sets of sealed sources.
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Elétrons , Radioisótopos de Gálio , Humanos , Tomografia por Emissão de Pósitrons , Mama , Mamografia , Imagens de FantasmasRESUMO
An efficient synthetic method for stereoselective construction of asymmetric quaternary carbon stereocenters, bearing nitrogen in the form of Boc-protected allyl amines, has been developed. This methodology is employed in the synthesis of marine alkaloids, manzacidin A and C.
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Carbono/química , Técnicas de Química Sintética/métodos , Nitrogênio/química , Pirimidinas/química , Pirimidinas/síntese química , Pirróis/química , Pirróis/síntese química , EstereoisomerismoRESUMO
X-ray and gamma-ray photons are widely used for imaging but require a mathematical reconstruction step, known as tomography, to produce cross-sectional images from the measured data. Theoretically, the back-to-back annihilation photons produced by positron-electron annihilation can be directly localized in three-dimensional space using time-of-flight information without tomographic reconstruction. However, this has not yet been demonstrated due to the insufficient timing performance of available radiation detectors. Here, we develop techniques based on detecting prompt Cerenkov photons, which when combined with a convolutional neural network for timing estimation resulted in an average timing precision of 32 picoseconds, corresponding to a spatial precision of 4.8 mm. We show this is sufficient to produce cross-sectional images of a positron-emitting radionuclide directly from the detected coincident annihilation photons, without using any tomographic reconstruction algorithm. The reconstruction-free imaging demonstrated here directly localizes positron emission, and frees the design of an imaging system from the geometric and sampling constraints that normally present for tomographic reconstruction.
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AIM: Cardiopulmonary resuscitation is vital for survival after cardiac arrest, and chest compressions are an important aspect of this. When performing chest compression in a hospital setting, the rescuer often has to kneel on the bed to overcome inconvenient differences in height between the rescuer and the bed. However, as yet no study has evaluated the quality of chest compressions in this position. The aim of this study was to examine the impact on the quality of chest compressions while kneeling on the bed. METHODS: Fifteen female students performed 2-min chest compressions on a manikin placed on the floor and a bed. Measurement parameters included compression depth, heart rate, integrated electromyogram, and a visual analog scale. The parameters were measured every 30 s and were statistically compared between the conditions. RESULTS: Compression depth at 30, 60, 90, and 120 s differed significantly between the conditions. Heart rate values at 150 and 210 s of recovery significantly differed between the conditions. Integrated electromyogram values for the trapezius, rectus femoris, and biceps femoris differed between the floor and bed conditions during 2-min chest compressions, whereas the external oblique muscle significantly differed at 60 and 120 s. Visual analog scales for fatigue, effectiveness, and stability significantly differed between the conditions. CONCLUSION: Kneeling on the bed does not enable grounding of the toe, causing the upper body to be unstable and limiting generation of the power required for chest compression. Our results suggest that rotation every minute is necessary to maintain effective cardiopulmonary resuscitation while kneeling on the bed.
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Reanimação Cardiopulmonar , Postura , Fadiga , Feminino , Frequência Cardíaca , Humanos , Masculino , Manequins , Adulto JovemRESUMO
A novel traceable point-like 68Ge/68Ga source was developed for calibration of positron emission tomography (PET) scanners. The source was equipped with a spherically symmetric acrylic positron absorber. The physical characteristics of photons emitted from the point-like source were evaluated by Monte Carlo simulation, considering possible geometrical uncertainties. The calibration factor of a clinical PET/CT scanner was evaluated using four manufactured point-like sources as a practical application of the point-like source. The results were consistent with that determined by the conventional cross-calibration method. The traceable point-like 68Ge/68Ga source is expected to be a simple and practical tool for determining the calibration factor and evaluating the physical characteristics of PET scanners.
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Radioisótopos de Gálio , Germânio , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/instrumentação , Radioisótopos , Calibragem , Método de Monte CarloRESUMO
Malaria causes a half a million deaths annually. The parasite intraerythrocytic lifecycle in the human bloodstream is the major cause of morbidity and mortality. Apical organelles of merozoite stage parasites are involved in the invasion of erythrocytes. A limited number of apical organellar proteins have been identified and characterized for their roles during erythrocyte invasion or subsequent intraerythrocytic parasite development. To expand the repertoire of identified apical organellar proteins we generated a panel of monoclonal antibodies against Plasmodium falciparum schizont-rich parasites and screened the antibodies using immunofluorescence assays. Out of 164 hybridoma lines, 12 clones produced monoclonal antibodies yielding punctate immunofluorescence staining patterns in individual merozoites in late schizonts, suggesting recognition of merozoite apical organelles. Five of the monoclonal antibodies were used to immuno-affinity purify their target antigens and these antigens were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Two known apical organelle protein complexes were identified, the high-molecular mass rhoptry protein complex (PfRhopH1/Clags, PfRhopH2, and PfRhopH3) and the low-molecular mass rhoptry protein complex (rhoptry-associated proteins complex, PfRAP1, and PfRAP2). A novel complex was additionally identified by immunoprecipitation, composed of rhoptry-associated membrane antigen (PfRAMA) and rhoptry neck protein 3 (PfRON3) of P. falciparum. We further identified a region spanning amino acids Q221-E481 within the PfRAMA that may associate with PfRON3 in immature schizonts. Further investigation will be required as to whether PfRAMA and PfRON3 interact directly or indirectly.
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Merozoítos , Plasmodium falciparum , Animais , Antígenos de Protozoários , Cromatografia Líquida , Eritrócitos , Humanos , Proteínas de Protozoários/genética , Espectrometria de Massas em TandemRESUMO
INTRODUCTION: To investigate the potential prognostic value of image analysis of pelvic bone metastasis in newly diagnosed prostate cancer patients. METHODS: Data from 69 patients with both bone scintigraphy and pelvic CT images were selected for this analysis. Open source software (3D Slicer version 4.8.1.) was used for image analysis. Metastatic pelvic bone lesions were manually contoured, and radiomic features were extracted. As risk factors for overall survival (OS) and cause-specific survival (CSS), 105 radiomic features and clinical risk factors including age, initial prostate-specific antigen, Gleason score, TNM stage, lactate dehydrogenase (LDH), hemoglobin (Hb), alkaline phosphatase, extent of disease, visceral metastases, and radiotherapy were assessed by uni- and multivariate analyses. RESULTS: Median follow-up was 41 months (range 0-157 months). Five-year overall survival and cause-specific survival rate were 37.9% and 43.5%, respectively. After multivariate analysis, LDH, Hb, and "maximum 2D diameter" defined as maximum tumor size in the axial plane were detected as risk factors for OS. Gleason sum, LDH, and maximum 2D diameter were detected as risk factors for CSS. CONCLUSION: Maximum 2D diameter was detected as a significant prognostic factor for metastatic prostate cancer patients.
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Neoplasias Ósseas/diagnóstico por imagem , Neoplasias Ósseas/secundário , Neoplasias da Próstata/mortalidade , Idoso , Idoso de 80 Anos ou mais , Seguimentos , Hemoglobinas/análise , Humanos , Processamento de Imagem Assistida por Computador , L-Lactato Desidrogenase/sangue , Masculino , Pessoa de Meia-Idade , Prognóstico , Neoplasias da Próstata/patologia , Cintilografia , Fatores de Risco , Tomografia Computadorizada por Raios XRESUMO
The energy loss distribution of beta+ particles is closely related to their maximum penetration depth distribution and annihilation point distribution. The latter is of practical importance for positron emission tomography. Experimental data related to the energy loss distribution are important for comprehensive validation of physics and simulation models of beta+ interactions. In this paper the authors present a new experimental approach that allows them to visually observe the beta+ energy loss distribution of a solution of nuclear medicine radioisotopes in a plastic scintillator using an optical camera. The authors also report a set of the first experimental results. A water solution of 18F was localized in a small hole in a plastic scintillator (BC430). Optical imaging of the scintillator yielded visual images of the energy loss distribution with a submillimeter resolution. The radial dependence in the energy distribution was quantitatively measured by analysis of the images, and exponential fitting parameters were obtained. The authors observed that the results of Monte Carlo simulation with EGS5 (version 1.0.2) and GEANT4 (version 4.9.01.p01) were consistent with those obtained experimentally. The results of the Monte Carlo simulation indicated that for a linear scale, the energy loss distribution in the scintillator was approximately the same as that in water, and the relative shape of the energy loss distribution was close to those of the maximum penetration depth distribution and annihilation point distribution. This paper also presents discussions about the further possibilities of this optical imaging approach. Thus, optical observation of the beta+ energy loss distribution in a scintillator is a promising technique for visual and quantitative experimental studies of beta+ emission from a solution of radioisotopes that are used in nuclear medicine.
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Partículas beta , Elétrons , Radioisótopos de Flúor/química , Dispositivos Ópticos , Imagens de Fantasmas , Água/química , Calibragem , Processamento de Imagem Assistida por Computador , Método de Monte Carlo , Plásticos , Tomografia por Emissão de Pósitrons , SoluçõesRESUMO
Proteins coating Plasmodium merozoite surface and secreted from its apical organelles are considered as promising vaccine candidates for blood-stage malaria. The rhoptry neck protein 12 of Plasmodium falciparum (PfRON12) was recently reported as a protein specifically expressed in schizonts and localized to the rhoptry neck of merozoites. Here, we assessed its potential as a vaccine candidate. We expressed a recombinant PfRON12 protein by a wheat germ cell-free system to obtain anti-PfRON12 antibody. Immunoblot analysis of schizont lysates detected a single band at approximately 40â¯kDa under reducing conditions, consistent with the predicted molecular weight. Additionally, anti-PfRON12 antibody recognized a single band around 80â¯kDa under non-reducing conditions, suggesting native PfRON12 forms a disulfide-bond-mediated multimer. Immunofluorescence assay and immunoelectron microscopy revealed that PfRON12 localized to the rhoptry neck of merozoites in schizonts and to the surface of free merozoites. The biological activity of anti-PfRON12 antibody was tested by in vitro growth inhibition assay (GIA), and the rabbit antibodies significantly inhibited merozoite invasion of erythrocytes. We then investigated whether PfRON12 is immunogenic in P. falciparum-infected individuals. The sera from P. falciparum infected individuals in Thailand and Mali reacted with the recombinant PfRON12. Furthermore, human anti-PfRON12 antibodies affinity-purified from Malian serum samples inhibited merozoite invasion of erythrocytes in vitro. Moreover, pfron12 is highly conserved with only 4 non-synonymous mutations in the coding sequence from approximately 200 isolates deposited in PlasmoDB. These results suggest that PfRON12 might be a potential blood-stage vaccine candidate antigen against P. falciparum.
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Antígenos de Protozoários/imunologia , Eritrócitos/parasitologia , Merozoítos/fisiologia , Plasmodium falciparum/imunologia , Proteínas de Protozoários/imunologia , Receptores Proteína Tirosina Quinases/imunologia , Animais , Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/genética , Infecções Assintomáticas/epidemiologia , Ensaio de Imunoadsorção Enzimática , Eritrócitos/imunologia , Imunofluorescência , Humanos , Vacinas Antimaláricas/genética , Vacinas Antimaláricas/imunologia , Malária Falciparum/epidemiologia , Malária Falciparum/imunologia , Malária Falciparum/parasitologia , Malária Falciparum/prevenção & controle , Mali/epidemiologia , Merozoítos/imunologia , Coelhos , Receptores Proteína Tirosina Quinases/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Esquizontes/química , Tailândia/epidemiologiaRESUMO
PURPOSE: To study the 3-year effect of oral nilvadipine, a calcium antagonist, on visual field performance and ocular circulation in open-angle glaucoma (OAG) with low-normal intraocular pressure (IOP). DESIGN: A randomized, placebo-controlled, double-masked, single-center trial. PARTICIPANTS: Patients with OAG who were younger than 65 years and had untreated IOP consistently of 16 mmHg or less. INTERVENTION: Oral nilvadipine (2 mg twice daily) or placebo was assigned randomly to patients fulfilling the criteria by the minimization method of balancing the groups according to age, refraction, and the mean deviation (MD) value (Humphrey Perimeter 30-2 SITA Standard Program; Humphrey Instruments, Inc., San Leandro, CA) of the eye with less negative MD. No topical ocular hypotensive drugs were prescribed. Visual field testing was performed every 3 months; fundus examination and IOP, blood pressure, and pulse rate measurements were carried out every month; and quantitative indexes of circulation in the optic disc rim (NB(ONH)) and choroid in the foveal area (NB(fovea)) were determined using the laser speckle method at 0, 3, 6, 12, 18, 24, 30, and 36 months. MAIN OUTCOME MEASURES: The time courses of MD, NB(ONH), and NB(fovea) in the eye with less negative MD. RESULTS: Thirty-three patients were enrolled; 17 were assigned to nilvadipine and 16 were assigned to placebo; 13 in each group completed the study. No significant intergroup difference was seen in age, refraction, or baseline values of any of the parameters. During the 3-year period, the IOP averaged 12.6 mmHg in the nilvadipine group and 12.8 mmHg in the placebo group (P>0.1), and no significant change from baseline or intergroup difference was seen in blood pressure or pulse rate. The estimated slope of change in the MD was less negative in the nilvadipine than in the placebo group (-0.01 vs. -0.27 decibels/year; P = 0.040). The NB(ONH) and NB(fovea) values remained increased compared with baseline for the study period by approximately 30% to 40% only in the nilvadipine group, and the intergroup difference was significant (P = 0.003 for NB(ONH) and P = 0.007 for NB(fovea)). CONCLUSIONS: Nilvadipine (2 mg twice daily) slightly slowed the visual field progression and maintained the optic disc rim, and the posterior choroidal circulation increased over 3 years in patients with OAG with low-normal IOP.
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Bloqueadores dos Canais de Cálcio/uso terapêutico , Corioide/irrigação sanguínea , Glaucoma de Ângulo Aberto/fisiopatologia , Pressão Intraocular/efeitos dos fármacos , Nifedipino/análogos & derivados , Disco Óptico/irrigação sanguínea , Campos Visuais/efeitos dos fármacos , Administração Oral , Pressão Sanguínea/efeitos dos fármacos , Método Duplo-Cego , Feminino , Glaucoma de Ângulo Aberto/tratamento farmacológico , Frequência Cardíaca/efeitos dos fármacos , Humanos , Masculino , Pessoa de Meia-Idade , Nifedipino/uso terapêutico , Fluxo Sanguíneo Regional/efeitos dos fármacosRESUMO
PURPOSE: Cherenkov radiation has recently received attention due to its prompt emission phenomenon, which has the potential to improve the timing performance of radiation detectors dedicated to positron emission tomography (PET). In this study, a Cherenkov-based three-dimensional (3D) position-sensitive radiation detector was proposed, which is composed of a monolithic lead fluoride (PbF2 ) crystal and a photodetector array of which the signals can be readout independently. METHODS: Monte Carlo simulations were performed to estimate the performance of the proposed detector. The position- and time resolution were evaluated under various practical conditions. The radiator size and various properties of the photodetector, e.g., readout pitch and single photon timing resolution (SPTR), were parameterized. The single photon time response of the photodetector was assumed to be a single Gaussian for the simplification. The photo detection efficiency of the photodetector was ideally 100% for all wavelengths. Compton scattering was included in simulations, but partly analyzed. To estimate the position at which a γ-ray interacted in the Cherenkov radiator, the center-of-gravity (COG) method was employed. In addition, to estimate the depth-of-interaction (DOI) principal component analysis (PCA), which is a multivariate analysis method and has been used to identify the patterns in data, was employed. The time-space distribution of Cherenkov photons was quantified to perform PCA. To evaluate coincidence time resolution (CTR), the time difference of two independent γ-ray events was calculated. The detection time was defined as the first photon time after the SPTR of the photodetector was taken into account. RESULTS: The position resolution on the photodetector plane could be estimated with high accuracy, by using a small number of Cherenkov photons. Moreover, PCA showed an ability to estimate the DOI. The position resolution heavily depends on the pitch of the photodetector array and the radiator thickness. If the readout pitch were ideally 0 and practically 3 mm, a full-width at half-maximum (FWHM) of 0.348 and 1.92 mm was achievable with a 10-mm-thick PbF2 crystal, respectively. Furthermore, first-order correlation could be observed between the primary principal component and the true DOI. To obtain a coincidence timing resolution better than 100-ps FWHM with a 20-mm-thick PbF2 crystal, a photodetector with SPTR of better than σ = 30 ps was necessary. CONCLUSIONS: From these results, the improvement of SPTR allows us to achieve CTR better than 100-ps FWHM, even in the case where a 20-mm-thick radiator is used. Our proposed detector has the potential to estimate the 3D interaction position of γ-rays in the radiator, using only time and space information of Cherenkov photons.
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Imageamento Tridimensional/instrumentação , Método de Monte Carlo , Tomografia por Emissão de Pósitrons/instrumentaçãoRESUMO
The jPET-D4 is the first PET scanner to introduce a unique four-layer depth-of-interaction (DOI) detector scheme in order to achieve high sensitivity and uniform high spatial resolution. This paper compares measurement and Monte Carlo simulation results of the static physics performance of this prototype research PET scanner. Measurement results include single and coincidence energy spectra, point and line source sensitivities, axial sensitivity profile (slice profile) and scatter fraction. We use GATE (Geant4 application for tomographic emission) as a Monte Carlo radiation transport model. Experimental results are reproduced well by the simulation model with reasonable assumptions on characteristic responses of the DOI detectors. In a previous study, the jPET-D4 was shown to provide a uniform spatial resolution as good as 3 mm (FHWM). In the present study, we demonstrate that a high sensitivity, 11.3 +/- 0.5%, is provided at the FOV centre. However, about three-fourths of this sensitivity is related to multiple-crystal events, for which some misidentification of the crystal cannot be avoided. Therefore, it is crucial to develop a more efficient way to identify the crystal of interaction and to reduce misidentification in order to make use of these high performance values simultaneously. We expect that effective sensitivity can be improved by replacing the GSO crystals with more absorptive crystals such as BGO and LSO. The results we describe here are essential to take full advantage of the next generation PET systems that have DOI recognition capability.
Assuntos
Interpretação de Imagem Assistida por Computador/métodos , Tomografia por Emissão de Pósitrons/instrumentação , Algoritmos , Desenho de Equipamento , Estudos de Avaliação como Assunto , Humanos , Método de Monte Carlo , Imagens de Fantasmas , Tomografia por Emissão de Pósitrons/métodos , Radiometria , Espalhamento de Radiação , Sensibilidade e Especificidade , SoftwareRESUMO
Genetic variability in Plasmodium falciparum malaria parasites hampers current malaria vaccine development efforts. Here, we hypothesize that to address the impact of genetic variability on vaccine efficacy in clinical trials, conserved antigen targets should be selected to achieve robust host immunity across multiple falciparum strains. Therefore, suitable vaccine antigens should be assessed for levels of polymorphism and genetic diversity. Using a total of one hundred and two clinical isolates from a region of high malaria transmission in Uganda, we analyzed extent of polymorphism and genetic diversity in four recently reported novel blood-stage malaria vaccine candidate proteins: Rh5 interacting protein (PfRipr), GPI anchored micronemal antigen (PfGAMA), rhoptry-associated leucine zipper-like protein 1 (PfRALP1) and Duffy binding-like merozoite surface protein 1 (PfMSPDBL1). In addition, utilizing the wheat germ cell-free system, we expressed recombinant proteins for the four candidates based on P. falciparum laboratory strain 3D7 sequences, immunized rabbits to obtain specific antibodies (Abs) and performed functional growth inhibition assay (GIA). The GIA activity of the raised Abs was demonstrated using both homologous 3D7 and heterologous FVO strains in vitro. Both pfripr and pfralp1 are less polymorphic but the latter is comparatively more diverse, with varied number of regions having insertions and deletions, asparagine and 6-mer repeats in the coding sequences. Pfgama and pfmspdbl1 are polymorphic and genetically diverse among the isolates with antibodies against the 3D7-based recombinant PfGAMA and PfMSPDBL1 inhibiting merozoite invasion only in the 3D7 but not FVO strain. Moreover, although Abs against the 3D7-based recombinant PfRipr and PfRALP1 proteins potently inhibited merozoite invasion of both 3D7 and FVO, the GIA activity of anti-PfRipr was much higher than that of anti-PfRALP1. Thus, PfRipr is regarded as a promising blood-stage vaccine candidate for next-generation vaccines against P. falciparum.