Discovery and Validation of Salivary Extracellular RNA Biomarkers for Noninvasive Detection of Gastric Cancer.

BACKGROUND: Biomarkers are needed for noninvasive early detection of gastric cancer (GC). We investigated salivary extracellular RNA (exRNA) biomarkers as potential clinical evaluation tools for GC. METHODS: Unstimulated whole saliva samples were prospectively collected from 294 individuals (163 GC and 131 non-GC patients) who underwent endoscopic evaluation at the Samsung Medical Center in Korea. Salivary transcriptomes of 63 GC and 31 non-GC patients were profiled, and mRNA biomarker candidates were verified with reverse transcription quantitative real-time PCR (RT-qPCR). In parallel, microRNA (miRNA) biomarkers were profiled and verified with saliva samples from 10 GC and 10 non-GC patients. Candidate biomarkers were validated with RT-qPCR in an independent cohort of 100/100 saliva samples from GC and non-GC patients. Validated individual markers were configured into a best performance panel. RESULTS: We identified 30 mRNA and 15 miRNA candidates whose expression pattern associated with the presence of GC. Among them, 12 mRNA and 6 miRNA candidates were verified with the discovery cohort by RT-qPCR and further validated with the independent cohort (n = 200). The configured biomarker panel consisted of 3 mRNAs (SPINK7, PPL, and SEMA4B) and 2 miRNAs (MIR140-5p and MIR301a), which were all significantly down-regulated in the GC group, and yielded an area under the ROC curve (AUC) of 0.81 (95% CI, 0.72-0.89). When combined with demographic factors, the AUC of the biomarker panel reached 0.87 (95% CI, 0.80-0.93). CONCLUSIONS: We have discovered and validated a panel of salivary exRNA biomarkers with credible clinical performance for the detection of GC. Our study demonstrates the potential utility of salivary exRNA biomarkers in screening and risk assessment for GC.

Discovery of a novel fibrous tissue in the spinal pia mater by polarized light microscopy.

Abstract It is very well known that spinal meninges are composed of three layers, dura, arachnoid and pia mater, and that the main components of pia mater are collagen and reticular fibers. However, the distribution of those fibers has not been extensively investigated but just described as a mesh of fibers. In this study, we detected novel structures, which are composed of unidirectionally arranged fibers, in a rat spinal pia mater by using a polarized light microscope. They were seen as three parallel lines, one of which ran along a posterior spinal vein and the rest two of which ran along a pair of posterior spinal arteries. Histological analysis including Masson's trichrome, picrosirius-red staining, Gordon & Sweet's staining and immunohistochemistry with anti-collagen type 1 and 3 antibodies uncovered that they are mainly composed of collagen fibers and some reticular fibers. In addition, a putative primo vessel was detected in the novel fibrous tissue, which was proven out to be different from a blood vessel. In conclusion, we report a newly detected fibrous structure in the spinal pia mater, which may contribute to provide tensile force to the spinal meninges and to harbor the primo vascular system inside.

Simplified image-based dosimetry using planar images and patient-specific S-values.

BACKGROUND: Single time point measurement approach and hybrid dosimetry were proposed to simplify the dosimetry process. It is anticipated that utilizing patient-specific S-value would enable more accurate dosimetry assessment based on imaging compared to using the conventional MIRD S-values. PURPOSE: We performed planar image-based dosimetry scaled with a single SPECT image for the entire treatment cycle using patient-specific S-values (PSS dosimetry) of organs. PSS dosimetry could further simplify the dosimetry procedure compared with a conventional 2D planar/3D SPECT hybrid dosimetry, as PSS dosimetry requires only one SPECT/CT image for the treatment of the entire cycle, whereas the conventional hybrid dosimetry requires a SPECT/CT image for each treatment cycle. METHODS: 177Lu-DOTATATE SPECT/CT and planar image datasets acquired from Seoul National University Hospital (SNUH, Seoul, Republic of Korea) were utilized for the evaluation. Images were acquired 4, 24, 48, and 120 h after patients' intravenous injection of 177Lu-DOTATATE. Dose estimations based on a Monte Carlo (MC) simulation using the Geant4 Application for Emission Tomography (GATE) (v.8.2) were considered as the reference. Planar image-based dosimetry scaled with a single SPECT image was performed using the patient-specific S-value (PSS). Briefly, the CT image was considered as the patient's anatomical reference and PSSs were quantified using the multiple voxel S-value (VSV) method. Then, PSS dosimetry was performed by obtaining activity information from sequential planar images and a scaling factor derived from a single SPECT/planar image pair. Hybrid dosimetry using sequential planar images and a single SPECT image was performed for comparison. The absorbed doses of the kidneys, bone marrow (BM) in the lumbar spine, liver, and spleen calculated using the PSS and hybrid dosimetries were compared with the reference MC results. RESULTS: The mean differences (MDs) of the self-absorption S-values between S-value of OLINDA/EXM and PSS for the kidneys, liver, and spleen were -0.04%, -2.39%, and -2.62%, respectively. However, the differences in the self-absorption S-values were significantly higher for the BM (84.99%) and the remainder of the body (ROB) (280.84%). The absorbed doses estimated by the PSS and hybrid dosimetries showed relatively high errors compared with MC simulation result, regardless of the organ. In contrast, the PSS and hybrid dosimetries produced similar dose estimates. For the entire cycles of the treatment, the MDs of absorbed doses between PSS and hybrid dosimetries were -3.31%, -6.04%, 3.37%, and -2.17% for the kidneys, BM, liver, and spleen, respectively. Through a correlation analysis and the Wilcoxon signed-rank test, we concluded that there was no significant difference between the results obtained by the two dosimetry methods. CONCLUSIONS: As the PSS was derived using CT images with actual anatomical information and organ-specific volume of interest (VOI), PSS dosimetry provided reliable results. PSS dosimetry was robust in estimating the absorbed dose for the later treatment cycles. Therefore, PSS dosimetry outperformed hybrid dosimetry in terms of dose estimation for a greater number of treatment cycles.

Mapping Korean National Health Insurance Claim Codes for Laboratory Test to SNOMED CT.

The aim of this study was to map Korean national health insurance claims codes for laboratory tests to SNOMED CT. The mapping source codes were 4,111 claims codes for laboratory test and mapping target codes were the International Edition of SNOMED CT released on July 31, 2020. We used rule-based automated and manual mapping methods. The mapping results were validated by two experts. Out of 4,111 codes, 90.5% were mapped to the concepts of procedure hierarchy in SNOMED CT. Of them, 51.4% of the codes were exactly mapped to SNOMED CT concepts, and 34.8% of the codes were mapped to SNOMED CT concepts as one-to-one mapping.

Voxel-Based Internal Dosimetry for 177Lu-Labeled Radiopharmaceutical Therapy Using Deep Residual Learning.

Purpose: In this study, we propose a deep learning (DL)-based voxel-based dosimetry method in which dose maps acquired using the multiple voxel S-value (VSV) approach were used for residual learning. Methods: Twenty-two SPECT/CT datasets from seven patients who underwent 177Lu-DOTATATE treatment were used in this study. The dose maps generated from Monte Carlo (MC) simulations were used as the reference approach and target images for network training. The multiple VSV approach was used for residual learning and compared with dose maps generated from deep learning. The conventional 3D U-Net network was modified for residual learning. The absorbed doses in the organs were calculated as the mass-weighted average of the volume of interest (VOI). Results: The DL approach provided a slightly more accurate estimation than the multiple-VSV approach, but the results were not statistically significant. The single-VSV approach yielded a relatively inaccurate estimation. No significant difference was noted between the multiple VSV and DL approach on the dose maps. However, this difference was prominent in the error maps. The multiple VSV and DL approach showed a similar correlation. In contrast, the multiple VSV approach underestimated doses in the low-dose range, but it accounted for the underestimation when the DL approach was applied. Conclusion: Dose estimation using the deep learning-based approach was approximately equal to that in the MC simulation. Accordingly, the proposed deep learning network is useful for accurate and fast dosimetry after radiation therapy using 177Lu-labeled radiopharmaceuticals.

Investigation of Electronic Signal Processing Chains for a Prototype TOF-PET System With 100-ps Coincidence Time Resolution.

We have evaluated CTR performance of four different mixed-signal front-end electronic readout configurations with the goal to achieve 100 picoseconds (ps) coincidence time resolution (CTR). The proposed TOF-PET detector elements are based on two 3 × 3 × 10 mm3 "fast LGSO" crystal segments, side-coupled to linear arrays of 3 × 3 mm2 silicon photomultipliers (SiPMs), to form a total crystal length of 20 mm. We studied multiple configurations and components for the front-end readout: 1) high speed radio frequency (RF) amplifiers; 2) an ASIC-based discriminator; 3) combination of RF amplifier, balun transformer, and discriminator ASIC; and 4) combination of balun transformer, and discriminator ASIC. Using two 3 × 3 × 10 mm3 fast LGSO crystals side coupled to a linear array of three SiPMs, coincidence data were experimentally acquired for each readout configuration in combination with a low jitter field programmable gate array (FPGA)-based time to digital converter (TDC). After evaluating timing performance of the three readout schemes, the best CTR value of 99.4 ± 1.9 ps FWHM was achieved for configuration (3), which is more than 20 ps better than the results achieved using configurations (1) and (2).

Comparison of voxel S-value methods for personalized voxel-based dosimetry of 177 Lu-DOTATATE.

PURPOSE: Voxel-based dosimetry is potentially accurate than organ-based dosimetry because it considers the anatomical variations in each individual and the heterogeneous radioactivity distribution in each organ. Here, voxel-based dosimetry for 177 Lu-DOTATATE therapy was performed using single and multiple voxel S-value (VSV) methods and compared with Monte Carlo simulations. To verify these methods, we adopted sequential 177 Lu-DOTATATE single-photon emission computed tomography and X-ray computed tomography (SPECT/CT) dataset acquired from Sunway Medical Centre using the major vendor's SPECT/CT scanner (Siemens Symbia Intevo). METHODS: The administered activity of 177 Lu-DOTATATE was 7.99 ± 0.36 GBq. SPECT/CT images were acquired 0.5, 4, 24, and 48 h after injection in Sunway Medical Centre. For the multiple VSV method, VSV kernels of 177 Lu in media with various densities were generated by Geant4 Application for Emission Tomography (GATE) simulation first. The second step involved the convolution of the time-integrated activity map with each kernel to produce medium-specific dose maps. Third, each medium-specific dose map was masked using binary medium masks, which were generated from CT-based density maps. Finally, all masked dose maps were summed to generate the final dose map. VSV methods with four different VSV sets (1, 4, 10, and 20 VSVs) were compared. Voxel-wise density correction for the single VSV method was also performed. The absorbed doses in the kidneys, bone marrow, and tumors were analyzed, and the relative errors between the VSV and Monte Carlo simulation approaches were estimated. Organ-based dosimetry using Organ Level INternal Dose Assessment/EXponential Modeling (OLINDA/EXM) was also compared. RESULTS: The accuracy of the multiple VSV approach increased with the number of dose kernels. The average dose estimation errors of a single VSV with density correction and 20 VSVs were less than 6% in most cases, although organ-based dosimetry using OLINDA/EXM yielded an error of up to 123%. The advantages of the single VSV method with density correction and the 20 VSVs over organ-based dosimetry were most evident in bone marrow and bone-metastatic tumors with heterogeneous medium properties. CONCLUSION: The single VSV method with density correction and multiple VSV method with 20 dose kernels enabled fast and accurate radiation dose estimation. Accordingly, voxel-based dosimetry methods can be useful for managing administration activity and for investigating tumor dose responses to further increase the therapeutic efficacy of 177 Lu-DOTATATE.

High-resolution time-of-flight PET detector with 100 ps coincidence time resolution using a side-coupled phoswich configuration.

Photon time-of-flight (TOF) capability in positron emission tomography (PET) enables reconstructed image signal-to-noise ratio (SNR) improvement. With the coincidence time resolution (CTR) of 100 picosecond (ps), a five-fold SNR improvement can be achieved with a 40 cm diameter imaging subject, relative to a system without TOF capability. This 100 ps CTR can be achieved for aclinically relevantdetector design (crystal element length ≥20 mm with reasonably high crystal packing fraction) using a side-readout PET detector configuration that enables 511 keV photon interaction depth-independent light collection efficiency and lower variance in scintillation photon transit time to the silicon photomultiplier (SiPM). In this study, we propose a new concept of TOF-PET detector to achieve high (<2 mm) resolution, using a 'side-coupled phoswich' configuration, where two crystals with different decay times (τd) are coupled in a side-readout configuration to a common row of photosensors. The proposed design was validated and optimized with GATE Monte Carlo simulation studies to determine an efficient detector design. Based on the simulation results, a proof-of-concept side-coupled phoswich detector design was developed comprising two LSO crystals with the size of 1.9 × 1.9 × 10 mm3with decay times of 34.39 and 43.07 ns, respectively. The phoswich crystals were side-coupled to the same three 4 × 4 mm2SiPMs and detector performances were evaluated. As a result of the experimental evaluation, the side-coupled phoswich configuration achieved CTR of 107 ± 3 ps, energy resolution of 10.5% ± 1.21% at 511 keV and >95% accuracy in identifying interactions in the two adjacent 1.9 × 1.9 × 10 mm3crystal elements using the time-over-threshold technique. Based on our results, we can achieve excellent spatial and energy resolution in addition to ∼100 ps CTR with this novel detector design.

Study of optical reflectors for a 100ps coincidence time resolution TOF-PET detector design.

Positron Emission Tomography (PET) reconstructed image signal-to-noise ratio (SNR) can be improved by including the 511 keV photon pair coincidence time-of-flight (TOF) information. The degree of SNR improvement from this TOF capability depends on the coincidence time resolution (CTR) of the PET system, which is essentially the variation in photon arrival time differences over all coincident photon pairs detected for a point positron source placed at the system center. The CTR is determined by several factors including the intrinsic properties of the scintillation crystals and photodetectors, crystal-to-photodetector coupling configurations, reflective materials, and the electronic readout configuration scheme. The goal of the present work is to build a novel TOF-PET system with 100 picoseconds (ps) CTR, which provides an additional factor of 1.5-2.0 improvement in reconstructed image SNR compared to state-of-the-art TOF-PET systems which achieve 225-400 ps CTR. A critical parameter to understand is the optical reflector's influence on scintillation light collection and transit time variations to the photodetector. To study the effects of the reflector covering the scintillation crystal element on CTR, we have tested the performance of four different reflector materials: Enhanced Specular Reflector (ESR) -coupled with air or optical grease to the scintillator; Teflon tape; BaSO4paint alone or mixed with epoxy; and TiO2paint. For the experimental set-up, we made use of 3 × 3 × 10 mm3fast-LGSO:Ce scintillation crystal elements coupled to an array of silicon photomultipliers (SiPMs) using a novel 'side-readout' configuration that has proven to have lower variations in scintillation light collection efficiency and transit time to the photodetector.Results: show CTR values of 102.0 ± 0.8, 100.2 ± 1.2, 97.3 ± 1.8 and 95.0 ± 1.0 ps full-width-half-maximum (FWHM) with non-calibrated energy resolutions of 10.2 ± 1.8, 9.9 ± 1.2, 7.9 ± 1.2, and 8.6 ± 1.7% FWHM for the Teflon, ESR (without grease), BaSO4(without epoxy) and TiO2paint treatments, respectively.

Simultaneous quantitative analysis of 3H and 14C radionuclides in aqueous samples via artificial neural network with a liquid scintillation counter.

Liquid scintillation counters are common instruments used in the measurement of pure beta-emitting radionuclides, and while they represent a conventional radiometric technique, they are still competitive for their potential to measure multiple radionuclides simultaneously. In this work, we propose an algorithm based on an artificial neural network (ANN) for the simultaneous analysis of the beta-ray spectra of 3H and 14C in dual beta-labeled samples using a liquid scintillation counter. We achieved percentage deviations below 5.0% using the proposed algorithm in 16 out of 18 cases, with RMSDs below 1.5% in 17 out of 18 cases. The trained ANN also produced activity ratios with high accuracy even while having to deal with highly fluctuating spectra. Results demonstrate that the rapid predictions with a short measurement time from our proposed ANN method are compatible with the calculated ones from previous studies that were obtained with long measurement times.

Scalable electronic readout design for a 100 ps coincidence time resolution TOF-PET system.

We have developed a scalable detector readout design for a 100 ps coincidence time resolution (CTR) time of flight (TOF) positron emission tomography (PET) detector technology. The basic scintillation detectors studied in this paper are based on 2 × 4 arrays of 3 × 3 × 10 mm3'fast-LGSO:Ce' scintillation crystals side-coupled to 6 × 4 arrays of 3 × 3 mm2silicon photomultipliers (SiPMs). We employed a novel mixed-signal front-end electronic configuration and a low timing jitter Field Programming Gate Array-based time to digital converter for data acquisition. Using a22Na point source, >10 000 coincidence events were experimentally acquired for several SiPM bias voltages, leading edge time-pickoff thresholds, and timing channels. CTR of 102.03 ± 1.9 ps full-width-at-half-maximum (FWHM) was achieved using single 3 × 3 × 10 mm3'fast-LGSO' crystal elements, wrapped in Teflon tape and side coupled to a linear array of 3 SiPMs. In addition, the measured average CTR was 113.4 ± 0.7 ps for the side-coupled 2 × 4 crystal array. The readout architecture presented in this work is designed to be scalable to large area module detectors with a goal to create the first TOF-PET system with 100 ps FWHM CTR.

Efficacy of voxel-based dosimetry map for predicting response to trans-arterial radioembolization therapy for hepatocellular carcinoma: a pilot study.

OBJECTIVE: Typical clinical dosimetry models for trans-arterial radioembolization (TARE) assume uniform dose distribution in each tissue compartment. We performed simple voxel-based dosimetry using post-treatment 90Y PET following TARE with 90Y-resin microspheres and investigated its prognostic value in a pilot cohort. METHOD: Ten patients with 14 hepatocellular carcinoma lesions who underwent TARE with 90Y-resin microspheres were retrospectively included. The partition model-based expected target tumor dose (TDp) was calculated using a pretreatment 99mTc-macroaggregated albumin scan. From post-treatment 90Y-microsphere PET and voxel-wise S-value kernels, voxel-based dose maps were produced and the absorbed dose of each lesion (TDv) was calculated. Heterogeneity of intratumoral absorbed doses was assessed using the SD and coefficient of variation of voxel doses. The response of each lesion was determined based on contrast-enhanced MRI or CT, or both. Lesion responses were classified as local control success or failure. Prognostic values of dosimetry parameters and clinicopathological factors were evaluated in terms of progression-free survival (PFS) of each lesion. RESULTS: TDv was significantly different between local control success and failure groups, whereas tumor size, TDp and intratumoral dose heterogeneity were not. Univariate survival analysis identified serum aspartate transaminase level ≥40 IU/L, tumor size ≥66 mm and TDv <81 Gy as significant prognostic factors for PFS. However, only TDv was an independent predictive factor in the multivariate analysis (P = 0.022). There was a significant correlation between TDv and PFS (P = 0.009; r = 0.669). CONCLUSIONS: In TARE, voxel-based dose index TDv can be estimated on post-treatment 90Y PET using a simple method. TDv was a more effective prognostic factor for TARE than TDp and clinicopathologic factors in this pilot study. Further studies are warranted on the role of voxel-based dose and dose distribution in TARE.

Dose-dependent pharmacokinetics of toxic metabolites is not related to increased toxicity following high-dose valproic acid in rats.

It has been reported that urinary excretion of two metabolites of valproic acid (VPA), 4-ene-valproic acid (4-VPA)and 2,4-diene-valproic acid (2,4-VPA), increased exponentially with the administration of high doses of VPA, and this increased formation of toxic metabolites could be related to VPA hepatotoxicity in humans. The aim of this study was to investigate whether the plasma level of 4-VPA and 2,4-VPA in rats corresponds to the urinary data for the same metabolites in humans.After the oral administration of VPA at doses of 20, 100 and 500 mg kg-1 in rats, the AUC0­24 h, 4-VPA/AUC0­24 h, VPA ratios (0.0399,0.0120 and 0.0100 for 20, 100 and 500 mg kg-1, respectively) and AUC0­24 h, 2,4-VPA/AUC0­24 h, VPA ratios (0.00104, 0.00201 and 0.00141, respectively) did not increase with increasing doses of VPA in rats. Thus, the plasma exposure of toxic metabolites normalized by dose remained unchanged (for 2,4-VPA) or even decreased (for 4-VPA) following high-dose VPA administration;this contradicts the findings of previous studies. Our results suggest that toxicity induced by high doses of VPA cannot be explained by a nonlinear increase of toxic metabolites in rats.

Preclinical Voxel-Based Dosimetry in Theranostics: a Review.

Due to the increasing use of preclinical targeted radionuclide therapy (TRT) studies for the development of novel theranostic agents, several studies have been performed to accurately estimate absorbed doses to mice at the voxel level using reference mouse phantoms and Monte Carlo (MC) simulations. Accurate dosimetry is important in preclinical theranostics to interpret radiobiological dose-response relationships and to translate results for clinical use. Direct MC (DMC) simulation is believed to produce more realistic voxel-level dose distribution with high precision because tissue heterogeneities and nonuniform source distributions in patients or animals are considered. Although MC simulation is considered to be an accurate method for voxel-based absorbed dose calculations, it is time-consuming, computationally demanding, and often impractical in daily practice. In this review, we focus on the current status of voxel-based dosimetry methods applied in preclinical theranostics and discuss the need for accurate and fast voxel-based dosimetry methods for pretherapy absorbed dose calculations to optimize the dose computation time in preclinical TRT.

Recovery of inter-detector and inter-crystal scattering in brain PET based on LSO and GAGG crystals.

Gadolinium aluminum gallium garnet (GAGG) is a promising scintillator crystal for positron emission tomography (PET) detectors owing to its advantages of energy resolution, light yield, and absence of intrinsic radiation. However, a large portion of the incident photons undergoes Compton scattering within GAGG crystal because of its low stopping power compared to that of lutetium-based crystals such as Lu2SiO5 (LSO). Inter-detector scattering (IDS) and inter-crystal scattering (ICS) result in loss of sensitivity and image quality of PET, respectively. We performed a Monte Carlo simulation study to evaluate IDS recovery in our currently developing brain-dedicated PET, and extended the idea to ICS recovery. We also compared the impact of the recoveries on LSO- and GAGG-based PET scanners. We measured the sensitivity and spatial resolution of the brain PET, and analyzed the image quality using a lesion phantom, a hot-rod phantom, and a 2D Hoffman phantom with applying IDS or ICS recovery. IDS recovery increased the PET sensitivity and improved the noise level of the reconstructed images. ICS recovery enhanced the spatial resolution and the contrast of the images was improved. As the occurrence rates of IDS and ICS were higher in GAGG than in LSO, the overall impact of IDS or ICS recovery was significant in GAGG. In conclusion, we showed that the proportional method would be suitable for IDS and ICS recoveries of PET, and emphasized the importance of ICS and IDS recoveries for PET using crystals with low stopping power.

Cost implications of adverse drug event-related emergency department visits - a multicenter study in South Korea.

Background: Adverse drug reactions (ADRs) increase health-care resource utilization, including that for emergency department (ED) visits. However, cost analyses of ADRs resulting in ED visits are scarce. Therefore, we aimed to estimate the direct medical costs before and after ADR occurrence and analyzed the cost-driving factors.Methods: The ADR cases were identified by a retrospective review of medical records of patients who visited the ED of three tertiary hospitals in South Korea from July to December 2014. The direct medical cost was estimated by the difference in costs six months before and after the ED visit. A generalized linear model was used to identify the ADR-associated cost-driving factors.Results: The mean cost per ADR increased by 26.1% (±SD = 4.3) during the six-month follow-up compared with that during the six months before the ED visit (p < 0.05). Preventable ADRs accounted for approximately 19.9% of the cost increase among all ADR cases. The regression analysis revealed that 'ADR-related hospitalization' was a significant (p < 0.05) factor leading to an increase in the direct medical costs.Conclusion: Drug-related ED visits increase the burden on health insurance systems and patients' out-of-pocket costs, mostly due to the hospitalization costs.

[18F]CB251 PET/MR imaging probe targeting translocator protein (TSPO) independent of its Polymorphism in a Neuroinflammation Model.

The 18 kDa translocator protein (TSPO) has been proposed as a biomarker for the detection of neuroinflammation. Although various PET probes targeting TSPO have been developed, a highly selective probe for detecting TSPO is still needed because single nucleotide polymorphisms in the human TSPO gene greatly affect the binding affinity of TSPO ligands. Here, we describe the visualization of neuroinflammation with a multimodality imaging system using our recently developed TSPO-targeting radionuclide PET probe [18F]CB251, which is less affected by TSPO polymorphisms. Methods: To test the selectivity of [18F]CB251 for TSPO polymorphisms, 293FT cells expressing polymorphic TSPO were generated by introducing the coding sequences of wild-type (WT) and mutant (Alanine → Threonine at 147th Amino Acid; A147T) forms. Competitive inhibition assay was conducted with [3H]PK11195 and various TSPO ligands using membrane proteins isolated from 293FT cells expressing TSPO WT or mutant-A147T, representing high-affinity binder (HAB) or low-affinity binder (LAB), respectively. IC50 values of each ligand to [3H]PK11195 in HAB or LAB were measured and the ratio of IC50 values of each ligand to [3H]PK11195 in HAB to LAB was calculated, indicating the sensitivity of TSPO polymorphism. Cellular uptake of [18F]CB251 was measured with different TSPO polymorphisms, and phantom studies of [18F]CB251-PET using 293FT cells were performed. To test TSPO-specific cellular uptake of [18F]CB251, TSPO expression was regulated with pCMV-TSPO (or shTSPO)/eGFP vector. Intracranial lipopolysaccharide (LPS) treatment was used to induce regional inflammation in the mouse brain. Gadolinium (Gd)-DOTA MRI was used to monitor the disruption of the blood-brain barrier (BBB) and infiltration by immune cells. Infiltration of peripheral immune cells across the BBB, which exacerbates neuroinflammation to produce higher levels of neurotoxicity, was also monitored with bioluminescence imaging (BLI). Peripheral immune cells isolated from luciferase-expressing transgenic mice were transferred to syngeneic inflamed mice. Neuroinflammation was monitored with [18F]CB251-PET/MR and BLI. To evaluate the effects of anti-inflammatory agents on intracranial inflammation, an inflammatory cytokine inhibitor, 2-cyano-3, 12-dioxooleana-1, 9-dien-28-oic acid methyl ester (CDDO-Me) was administered in intracranial LPS challenged mice. Results: The ratio of IC50 values of [18F]CB251 in HAB to LAB indicated similar binding affinity to WT and mutant TSPO and was less affected by TSPO polymorphisms. [18F]CB251 was specific for TSPO, and its cellular uptake reflected the amount of TSPO. Higher [18F]CB251 uptake was also observed in activated immune cells. Simultaneous [18F]CB251-PET/MRI showed that [18F]CB251 radioactivity was co-registered with the MR signals in the same region of the brain of LPS-injected mice. Luciferase-expressing peripheral immune cells were located at the site of LPS-injected right striatum. Quantitative evaluation of the anti-inflammatory effect of CDDO-Me on neuroinflammation was successfully monitored with TSPO-targeting [18F]CB251-PET/MR and BLI. Conclusion: Our results indicate that [18F]CB251-PET has great potential for detecting neuroinflammation with higher TSPO selectivity regardless of polymorphisms. Our multimodal imaging system, [18F]CB251-PET/MRI, tested for evaluating the efficacy of anti-inflammatory agents in preclinical studies, might be an effective method to assess the severity and therapeutic response of neuroinflammation.

The relationship between glucuronide conjugate levels and hepatotoxicity after oral administration of valproic acid.

The aim of this study was to investigate the relationship between hepatotoxicity, levels of glucuronide conjugates of valproic acid (VPA), and the toxic metabolites of VPA (4-ene VPA and 2,4-diene VPA). We also examined whether hepatotoxicity could be predicted by the urinary excretion levels of VPA and its toxic metabolites. VPA was administrated orally in rats in amounts ranging from 20 mg/kg to 500 mg/kg. Free and total (free plus glucuronide conjugated) VPA, 4-ene VPA, and 2,4-diene VPA were quantified in urine and liver using gas chromatography-mass spectrometry. Serum levels of aspartate aminotransferase, alanine aminotransferase, and alpha-glutathione S-transferase (alpha-GST) were also determined to measure the level of hepatotoxicity. The serum alpha-GST level increased slightly at the 20 mg/kg dose, and substantially increased at the 100 and 500 mg/kg dose; aspartate aminotransferase and alanine aminotransferase levels did not change with the administration of increasing doses of VPA. The liver concentration of free 4-ene VPA and the urinary excretion of total 4-ene VPA were the only measures that correlated with the increase in the serum alpha-GST level (p < 0.094 and p < 0.023 respectively). From these results, we conclude that hepatotoxicity of VPA correlates with liver concentration of 4-ene VPA and can be predicted by the urinary excretion of total 4-ene VPA.

Metabolomics study with gas chromatography-mass spectrometry for predicting valproic acid-induced hepatotoxicity and discovery of novel biomarkers in rat urine.

Three different doses of valproic acid (20, 100, and 500 mg/kg/d) are administered orally to Sprague-Dawley rats for 5 days, and the feasibility of metabolomics with gas chromatography-mass spectrometry as a predictor of the hepatotoxicity of valproic acid is evaluated. Body weight is found to decrease with the 100-mg/kg/d dose and significantly decrease with the 500-mg/kg/d dose. Mean excreted urine volume is lowest in the 500-mg/kg/d group among all groups. The plasma level of alpha-glutathione-S-transferase, a sensitive and earlier biomarker for hepatotoxicity, increases significantly with administration of 100 and 500 mg/kg/d; however, there is not a significant difference in alpha-glutathione-S-transferase plasma levels between the control and 20-mg/kg/d groups. Clusters in partial least squares discriminant analysis score plots show similar patterns, with changes in physiological conditions and plasma levels of alpha-glutathione-S-transferase; the cluster for the control and 20-mg/kg/d groups does not clearly separate, but the clusters are separate for 100- and 500-mg/kg/d groups. A biomarker of hepatotoxicity, 8-hydroxy-2'-deoxyguanosine and octanoylcarnitine, is identified from nontargeted and targeted metabolic profiling. These results validate that metabolic profiling using gas chromatography-mass spectrometry could be a useful tool for finding novel biomarkers. Thus, a nontargeted metabolic profiling method is established to evaluate the hepatotoxicity of valproic acid and demonstrates proof-of-concept that metabolomic approach with gas chromatography-mass spectrometry has great potential for predicting valproic acid-induced hepatotoxicity and discovering novel biomarkers.

Deep-dose: a voxel dose estimation method using deep convolutional neural network for personalized internal dosimetry.

Personalized dosimetry with high accuracy is crucial owing to the growing interests in personalized medicine. The direct Monte Carlo simulation is considered as a state-of-art voxel-based dosimetry technique; however, it incurs an excessive computational cost and time. To overcome the limitations of the direct Monte Carlo approach, we propose using a deep convolutional neural network (CNN) for the voxel dose prediction. PET and CT image patches were used as inputs for the CNN with the given ground truth from direct Monte Carlo. The predicted voxel dose rate maps from the CNN were compared with the ground truth and dose rate maps generated voxel S-value (VSV) kernel convolution method, which is one of the common voxel-based dosimetry techniques. The CNN-based dose rate map agreed well with the ground truth with voxel dose rate errors of 2.54% ± 2.09%. The VSV kernel approach showed a voxel error of 9.97% ± 1.79%. In the whole-body dosimetry study, the average organ absorbed dose errors were 1.07%, 9.43%, and 34.22% for the CNN, VSV, and OLINDA/EXM dosimetry software, respectively. The proposed CNN-based dosimetry method showed improvements compared to the conventional dosimetry approaches and showed results comparable with that of the direct Monte Carlo simulation with significantly lower calculation time.