Generation of a human SLC12A3 knock-in human induced pluripotent stem cell line (CMCi014-A-82) using CRISPR-Cas9 system.

Gitelman's disease is caused by a genetic mutation in the solute carrier family 12 member 3 (SLC12A3) gene, which encodes the sodium chloride cotransporter. In this study, we generated a stable human induced pluripotent stem cell (hiPSC) line, WTC11-SLC12A3 (CMCi014-A-82), by knocking in the entire SLC12A3 gene at the SHS231 locus in healthy wild-type control hiPSCs (WTC11). We verified that WTC11-SLC12A3 expressed pluripotency markers and exhibited normal stem cell morphology. Furthermore, this cell line maintains a normal karyotype and can differentiate into the three germ layers. Therefore, this cell line may provide a basis for gene therapy for Gitelman's disease.

Enhanced toxicity-free astaxanthin extraction from Haematococcus pluvialis via concurrent cell disruption and demulsification.

The extraction of astaxanthin from Haematococcus pluvialis involves the utilization of petroleum-derived organic solvents or supercritical CO2, beset by safety concerns, high costs, and environmental sustainability limitations. This study, in contrast, employed a method involving the adjustment of salt concentration, propylene glycol, and vegetable oil fraction to disrupt emulsion in aqueous cell lysates for facilitating the separation of astaxanthin. Under optimized conditions, an astaxanthin-containing oil with a content of 1.88% was obtained even with the use of wet biomass, and four rounds of consecutive extraction resulted in a cumulative recovery yield of 66.41%. This process produced astaxanthin-enriched soybean oil with 9.49 times improved antioxidant capacity that satisfies a requirement for health functional application. Omitting the solvent removal and drying processes, which consume tremendous energy, can reduce the production cost by 2.98 times compared to conventional methods. Consequently, this study suggests an effective technique for producing edible oil containing H. pluvialis-derived astaxanthin.

Relationship between rheological properties, sensory perception, and overall acceptability of thickened liquids for dysphagia in young and old healthy individuals.

This study investigated the relationship between rheological properties, sensory perception, and overall acceptability in healthy young and old groups for dysphagia thickened liquids. Unflavored (UTL) and flavored (FTLP) thickened liquids were prepared using tap water or pomegranate juice at 10 different viscosity levels. The rheological properties were then evaluated via syringe flow test and line spread test (LST). When the apparent viscosity levels of UTL and FTLP were similar, the syringe test and LST results were also similar, indicating consistent flow behavior. Sensory perception evaluations showed that the young group better distinguished viscosity differences between stages compared to the old group. Regarding overall acceptability, the old group preferred samples with higher apparent viscosity than the young group. Principal component analysis and k-means cluster analysis were used to explore correlations between variables and classify thickened liquids into four groups. This can serve the foundation for standardized texture grades of dysphagia thickened liquids, considering rheological characteristics and sensory profiles.

Increased Multiplexity in Optical Tissue Clearing-Based Three-Dimensional Immunofluorescence Microscopy of the Tumor Microenvironment by Light-Emitting Diode Photobleaching.

Optical tissue clearing and three-dimensional (3D) immunofluorescence (IF) microscopy is transforming imaging of the complex tumor microenvironment (TME). However, current 3D IF microscopy has restricted multiplexity; only 3 or 4 cellular and noncellular TME components can be localized in cleared tumor tissue. Here we report a light-emitting diode (LED) photobleaching method and its application for 3D multiplexed optical mapping of the TME. We built a high-power LED light irradiation device and temperature-controlled chamber for completely bleaching fluorescent signals throughout optically cleared tumor tissues without compromise of tissue and protein antigen integrity. With newly developed tissue mounting and selected region-tracking methods, we established a cyclic workflow involving IF staining, tissue clearing, 3D confocal microscopy, and LED photobleaching. By registering microscope channel images generated through 3 work cycles, we produced 8-plex image data from individual 400 µm-thick tumor macrosections that visualize various vascular, immune, and cancer cells in the same TME at tissue-wide and cellular levels in 3D. Our method was also validated for quantitative 3D spatial analysis of cellular remodeling in the TME after immunotherapy. These results demonstrate that our LED photobleaching system and its workflow offer a novel approach to increase the multiplexing power of 3D IF microscopy for studying tumor heterogeneity and response to therapy.

Comparison of microbial diversity and metabolites on household and commercial doenjang.

In this study, the microbial diversity, free amino acid (FAA), and biological activities of household doenjang (HDJ) from four different regions and commercial doenjang (CDJ) four manufacturers were analyzed. And volatile flavor compound (VFC) and isoflavone profiles were analyzed using gas chromatograph-mass spectrometer (GC-MS) and high performance liquid chromatography (HPLC), respectively. The major bacterial genus in 1HDJ and 2HDJ was Bacillus (97.5%), while in 3HDJ and 4HDJ, it was Enterobacter (47.5%) and Pseudomonas (80%), respectively. Tetragenococcus was the main bacterial genus of CDJ. The Zygosaccharomyces genus among yeast was comparatively high in all samples. In all samples, glutamic acid predominated among the FAAs, and the 3-methyl butanal, benzeneacetaldehyde, and diallyl disulphide were major VFCs. CDJ contained higher levels of isoflavone-glycoside and total phenolics. Except for 3HDJ and 4CDJ, the isoflavone-aglycone and total flavonoid contents were higher in HDJ. The correlation between bacterial genus and metabolited of doenjang showed that Tetragenococcus was closely related to glutamic acid, Bacillus was related to aglycones and ammonia, and Pseudomonas was highly related to isovaleric acid. While, correlation between yeast genus and metabolited of doenjang confirmed that Candida, Hanseniaspora, and Saccharomyces were related with furfural, benzeneacetaldehyde, and 3-methyl butanal, respectively. The results of this study can be utilized as basic data for the industrialization and development of doenjang.

Changes in walking practice and associated factors during the coronavirus disease 2019 pandemic.

The coronavirus disease 2019 (COVID-19) pandemic has induced many lifestyle changes, with reductions in healthy activities, including walking. The aim of this cross-sectional study was to identify strategies to maintain good health during a pandemic. Demographics, walking habits, health behaviors, health status, recognition of social capital, and changes in walking practice of 800 residents were evaluated using a structured questionnaire. Among the participants, 73.9%, 15.8%, and 10.4% reported no change, a decrease, and an increase in walking practice, respectively. Individuals in their 50 s and 60 s showed 6.388- and 5.054-times greater increases in walking, respectively, than did those aged 19-29 years. Walking increased more among high-income people than among low-income people. Additionally, walking practice increased in people participating in social gatherings, those with pets that needed walks, those with a COVID-19 history, those who considered walking as both a habitual and leisure activity, those with nearly no stress, and those satisfied with their safety, the natural environment, and public transportation; other health behaviors or conditions were not related to an increase in walking practice. To maintain good health during a pandemic, walking practice should be promoted in people in their 20 s and low-income groups. Moreover, people should be encouraged to attend meetings, including online interaction, and safe walking routes should be created within the community. Walking should be encouraged as a method of relieving stress, and the benefits of walking for recovery after an infectious disease should be emphasized. Furthermore, the direct effects of walking should be studied.

Report from an online workshop on family physician socialization: Reaffirmation using a hypothetical case.

This letter provides a report of our online workshop on the professional socialization of family physicians attended by family medicine residents and staff doctors in Japan. Participants engaged in small group discussions after reading hypothetical cases that threatened the identity of family physicians. Participants considered the importance of reaffirming and improving their strengths during discussions.

Electrocardiographic P terminal force in lead V1, its components, and the association with stroke and atrial fibrillation or flutter.

BACKGROUND: The electrocardiographic (ECG) marker P terminal force V1 (PTFV1) is generally perceived as a marker of left atrial pathology and has been associated with atrial fibrillation or flutter (AF). OBJECTIVE: The purpose of this study was to determine the association between PTFV1 components (duration and amplitude) and incident AF and stroke/transient ischemic attack (TIA). METHODS: The study included patients with an ECG recorded at the Copenhagen General Practitioners Laboratory in 2001 to 2011. PTFV1 ≥4 mV·ms was considered abnormal. Patients with abnormal PTFV1 were stratified into tertiles based on duration (PTDV1) and amplitude (PTAV1) values. Cox regressions adjusted for age, sex, and relevant comorbidities were used to investigate associations between abnormal PTFV1 components and AF and stroke/TIA. RESULTS: Of 267,636 patients, 5803 had AF and 18,176 had stroke/TIA (follow-up 6.5 years). Abnormal PTFV1 was present in 44,549 subjects (16.7%) and was associated with an increased risk of AF and stroke/TIA. Among patients with abnormal PTFV1, the highest tertile of PTDV1 (78-97 ms) was associated with the highest risk of AF (hazard ratio [HR] 1.37; 95% confidence interval [CI] 1.23-1.52) and highest risk of stroke/TIA (HR 1.13; 95% CI 1.05 -1.20). For PTAV1, the highest tertile (78-126 µV) conferred the highest risk of AF and stroke/TIA (HR 1.20; 95% CI 1.09-1.32; and HR 1.21; 95% CI 1.14-1.25, respectively). CONCLUSION: Abnormal PTFV1 was associated with an increased risk of AF and stroke/TIA. Increasing PTDV1 showed a dose-response relationship with the development of AF and stroke/TIA, whereas the association between PTAV1 and AF was less apparent.

Humic-like crop stimulatory activities of coffee waste induced by incorporation of phytotoxic phenols in melanoidins during coffee roasting: Linking the Maillard reaction to humification.

Here we showed that the water-soluble components of fresh green coffee beans inhibit the growth of lettuce in hydroponic systems, whereas those of roasted coffee waste facilitate it. The growth enhancement was hardly related to hydroponic parameters (i.e., pH and electric conductivity) or the nitrogen contents of the extracts. Rather, the presence of chromogenic polymeric melanoidins in the coffee waste was found to be crucial for the crop growth acceleration. The quantitative comparison of low-molecular-weight organics including phytotoxic phenolics between the extracts suggested that Maillard reactions occurring during coffee roasting transform the phenolics into polymeric melanoidin products. The identification of humic-like molecular compositions in the roasted coffee waste and the restoration of crop-stimulating activity by the addition of a phenol oxidase to the fresh coffee bean extract also supported that the low-molecular-weight phenols are oxidatively coupled during the roasting, which was consistent with the bottom-up synthesis of crop-stimulatory humic substances.

Color Matching Generation Algorithm for Animation Characters Based on Convolutional Neural Network.

In recent years, for China, animation industry is a relatively new and mature emerging national sunrise industry after animation industry, which appears on the world stage more and more frequently and is widely concerned and valued by people from all over the world. Therefore, this paper innovatively uses the convolutional neural network algorithm to innovate the color matching generation of animation characters and improve the traditional technology of color matching for animation characters. In this paper, we mainly use Generative Adversarial Network (GAN), Deep Convolutional Generative Adversarial Network and VGG model, and multiscale discriminator theory and use ACGAN research method. And we study this paper's innovative LMV-ACGAN research method, and we have come to the conclusion that other models have higher collapse rate than this model; this model has higher color matching of anime characters. Color matching improves with the increase of convolutional neural network utilization, etc. Moreover, superior and minor reviews of this study are provided to make later researchers understand this study more rationally and objectively.

Integration of multiplatform metabolomics and multivariate analysis for geographical origin discrimination of Panax ginseng.

As the health food industry grows, the market for ginseng also expands globally. Korean ginseng (Panax ginseng C. A. Meyer) is cultivated in East Asian countries such as Korea, China, and Japan. The metabolic profile of plants can vary depending on the cultivation environment. As such, in this study, we aimed to compare the differences in the metabolic profiles of P. ginseng cultivated in Korea, China, and Japan, and to construct a library of these metabolite data. Using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS), we profiled 62 types of secondary metabolites, ginsenosides, using optimized analysis conditions to separate peaks with a high-resolution for about 30 min. In addition, we selected ginsenosides showing differences between their origins were selected among the possible origins in the S-line plot of orthogonal partial least squares discriminant analysis (OPLS-DA), which we quantitatively analyzed using UPLC-tandem mass spectrometry (UPLC-MS/MS). The contents of ginsenosides Ra1, Ra2, Ra3, and Rk1 were high in Korean P. ginseng; the contents of ginsenosides Rb1, Rb2, and Rc were high in Japanese P. ginseng; and the contents of the ginsenoside Ro was high in Chinese P. ginseng. We also analyzed the primary metabolite contents using nuclear magnetic resonance (NMR) spectroscopy and gas chromatography time-of-flight mass spectrometry (GC-TOF/MS). Japanese P. ginseng showed a high sucrose content and Korean P. ginseng showed high contents of most amino acids and organic acids. In the PLS-DA results of multivariate statistical analysis using the data obtained from each analysis instrument, we observed a clear clustering among the three origins. Although a genetically identical species, the metabolic profile substantially differs depending on the cultivation environment. Because ginsenoside, having many biological activities, showed origin-dependent origins, when P. ginseng is used for medicinal purposes, its content by origin should be considered. After disclosing the profiling results of these metabolites, we expect that they will be used in future ginseng research.

Topology dependent modification of layered double hydroxide for therapeutic and diagnostic platform.

Layered double hydroxide is a family of two-dimensional materials with wide range of compositions. Recently, its ability to accommodate various chemical species and biocompatibility have been attracted in the biomedical applications to develop drug delivery system and nanodiagnostics. In this review, we categorized biomedical approaches of layered double hydroxide with respect to the three topologies of, namely, interlayer space, outer surface with particle edge, and the lattice points. There have been extensive researches on the intercalation of drug or tracing to make use of interlayer space of layered double hydroxide for drug stabilization, sustained release, cellular delivery and etc. Outer surface or edge has been utilized to immobilization of large therapeutic moieties and to attach tracing moiety. Lattice points consisting of various metal species could be utilized for the specific metal species like paramagnetic elements or radioisotopes. Based on these topologies in layered double hydroxide, both the synthetic routes and the achieved functionalities in terms of biomedical application will be discussed.

Pembrolizumab versus paclitaxel for previously treated advanced gastric or gastroesophageal junction cancer (KEYNOTE-063): A randomized, open-label, phase 3 trial in Asian patients.

BACKGROUND: KEYNOTE-063 (NCT03019588) investigated pembrolizumab versus paclitaxel as second-line therapy in Asian patients with advanced programmed death ligand 1 (PD-L1)-positive (combined positive score ≥1) gastric/gastroesophageal junction (GEJ) cancer. METHODS: This randomized, open-label, phase 3 study was conducted at 36 medical centers in China (mainland), Malaysia, South Korea, and Taiwan. Patients were randomly assigned 1:1 to 200 mg of pembrolizumab intravenously every 3 weeks for ≤2 years or 80 mg/m2 of paclitaxel intravenously every week. Primary end points were overall survival (OS) and progression-free survival (PFS). Secondary end points were objective response rate (ORR) per Response Evaluation Criteria in Solid Tumors version 1.1 and safety. RESULTS: Between February 16, 2017, and March 12, 2018, 94 patients were randomly assigned (47 pembrolizumab/47 paclitaxel) after screening; enrollment was stopped on March 12, 2018, based on the results of the global KEYNOTE-061 study, and patients were followed until the last patient's last visit. Median OS was 8 months (95% confidence interval [CI], 4-10 months) with pembrolizumab versus 8 months (95% CI, 5-11 months) with paclitaxel (hazard ratio [HR], 0.99; 95% CI, 0.63-1.54). Median PFS was 2 months (95% CI, 1-3 months) with pembrolizumab versus 4 months (95% CI, 3-6 months) with paclitaxel (HR, 1.62; 95% CI, 1.04-2.52). ORR was 13% for pembrolizumab versus 19% for paclitaxel. Any-grade treatment-related adverse events occurred in 28 pembrolizumab-treated patients (60%) and 42 paclitaxel-treated patients (96%); grades 3 to 5 events occurred in 5 patients (11%) and 28 patients (64%), respectively. CONCLUSIONS: Definitive conclusions about the efficacy of second-line pembrolizumab in Asian patients with advanced PD-L1-positive gastric/GEJ cancer are limited because of insufficient power, but pembrolizumab was well tolerated in this patient population. Efficacy followed a trend similar to that observed in the phase 3 KEYNOTE-061 trial.

Intramolecular N-H⋠⋠⋠F Hydrogen Bonding Interaction in a Series of 4-Anilino-5-Fluoroquinazolines: Experimental and Theoretical Characterization of Electronic and Conformational Effects.

The 4-anilino-6,7-ethylenedioxy-5-fluoroquinazoline scaffold is presented as a novel model system for the characterization of the weak NH⋅⋅⋅F hydrogen bonding (HB) interaction. In this scaffold, the aniline NH proton is forced into close proximity with the nearby fluorine (dH,F ∼2.0 Å, ∠∼138°), and a through-space interaction is observed by NMR spectroscopy with couplings (1h JNH,F ) of 19±1 Hz. A combination of experimental (NMR spectroscopy and X-ray crystallography) and theoretical methods (DFT calculations) were used for the characterization of this weak interaction. In particular, the effects of conformational rigidity and steric compression on coupling were investigated. This scaffold was used for the direct comparison of fluoride with methoxy as HB acceptors, and the susceptibility of the NH⋅⋅⋅F interaction to changes in electron distribution and resonance was probed by preparing a series of molecules with different electron-donating or -withdrawing groups in the positions para to the NH and F. The results support the idea that fluorine can act as a weak HB acceptor, and the HB strength can be modulated through additive and linear electronic substituent effects.

Association Between Longitudinal Blood Pressure Trajectory and the Progression of Chronic Kidney Disease: Results From the KNOW-CKD.

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Risk factors for the development of post-infectious bronchiolitis obliterans after Mycoplasma pneumoniae pneumonia in the era of increasing macrolide resistance.

BACKGROUND: The prevalence of macrolide-resistant Mycoplasma pneumoniae (MP) pneumonia has been rapidly increased. MP pneumonia is a risk factor for the development of post-infectious bronchiolitis obliterans (PIBO). The aim of the present study was to identify the risk factors for the development of PIBO after MP pneumonia in the era of increasing macrolide resistance of MP. MATERIALS AND METHODS: This retrospective study enrolled 150 children with a mean age of 6.0 years admitted to the hospital due to MP pneumonia between May 2019 and February 2020 at a tertiary hospital. The clinical, radiologic, and laboratory data were obtained using retrospective chart review. RESULTS: Eighteen children (12%) were diagnosed with PIBO after MP pneumonia. PIBO was diagnosed after a mean duration of 100.0 days (range, 6-268 days) from symptom onset. The respiratory virus co-infection (adjusted odds ratio [aOR], 4.069; 95% confidence interval [95% CI], 1.224-13.523), adenovirus co-infection (aOR, 5.607; 95% CI, 1.801-17.454), longer duration between symptom onset and admission (aOR, 1.150; 95% CI, 1.020-1.298), higher levels of serum lactate dehydrogenase (LDH) at the time of admission (aOR, 1.001; 95% CI, 1.000-1.003), and poor response to stepwise treatment increased the risk for development of PIBO after MP pneumonia. However, macrolide resistance of MP was not associated with development of PIBO after MP pneumonia. CONCLUSION: The present study suggests that respiratory virus co-infection, including adenovirus, poor response to the treatment of MP pneumonia, and higher levels of serum LDH, but not macrolide resistance of MP, are risk factors of PIBO after MP pneumonia.

Engineering Core-Shell Structures of Magnetic Ferrite Nanoparticles for High Hyperthermia Performance.

Magnetic ferrite nanoparticles (MFNs) with high heating efficiency are highly desirable for hyperthermia applications. As conventional MFNs usually show low heating efficiency with a lower specific loss power (SLP), extensive efforts to enhance the SLP of MFNs have been made by varying the particle compositions, sizes, and structures. In this study, we attempted to increase the SLP values by creating core-shell structures of MFNs. Accordingly, first we synthesized three different types of core ferrite nanoparticle of magnetite (mag), cobalt ferrite (cf) and zinc cobalt ferrite (zcf). Secondly, we synthesized eight bi-magnetic core-shell structured MFNs; Fe3O4@CoFe2O4 (mag@cf1, mag@cf2), CoFe2O4@Fe3O4 (cf@mag1, cf@mag2), Fe3O4@ZnCoFe2O4 (mag@zcf1, mag@zcf2), and ZnCoFe2O4@Fe3O4 (zcf@mag1, zcf@mag2), using a modified controlled co-precipitation process. SLP values of the prepared core-shell MFNs were investigated with respect to their compositions and core/shell dimensions while varying the applied magnetic field strength. Hyperthermia properties of the prepared core-shell MFNs were further compared to commercial magnetic nanoparticles under the safe limits of magnetic field parameters (<5 × 109 A/(m·s)). As a result, the highest SLP value (379.2 W/gmetal) was obtained for mag@zcf1, with a magnetic field strength of 50 kA/m and frequency of 97 kHz. On the other hand, the lowest SLP value (1.7 W/gmetal) was obtained for cf@mag1, with a magnetic field strength of 40 kA/m and frequency of 97 kHz. We also found that magnetic properties and thickness of the shell play critical roles in heating efficiency and hyperthermia performance. In conclusion, we successfully enhanced the SLP of MFNs by engineering their compositions and dimensions.

Comparison of the performance of risk scoring systems for tumor bleeding in patients with inoperable gastric cancer.

BACKGROUND: The Glasgow - Blatchford bleeding score (GBS) and admission and full Rockall scores are widely used risk scoring systems to stratify risk and determine the need for intervention in patients with upper gastrointestinal bleeding. We evaluated the performance of these risk scoring systems in patients with gastric cancer and tumor bleeding. METHODS: This retrospective study included patients with inoperable gastric cancer who presented with tumor bleeding at the National Cancer Center, Korea, between 2001 and 2015. The GBS, and admission and full Rockall scores were calculated. Primary outcome was the performance of the risk scoring systems in predicting the need for urgent interventions (endoscopic therapy, transarterial embolization, and surgery). The risk factors associated with urgent intervention were analyzed. RESULTS: Of 357 patients with tumor bleeding, 118 (33.1 %; 116 endoscopic therapy, 2 trans-arterial embolization) required urgent intervention. The full Rockall score was better at predicting the need for urgent intervention (area under the receiver operating characteristic curve = 0.78; P < 0.001) than the GBS (0.56) and admission Rockall score (0.56). Hemostatic intervention was not performed in patients with a full Rockall score ≤ 6 (103 patients, 28.9 %). On multivariate analysis, endoscopic stigmata of recent hemorrhage of Forrest classes Ia - IIb were significant factors associated with urgent intervention. CONCLUSIONS: The full Rockall score was superior to the GBS and admission Rockall score in predicting the need for urgent intervention for tumor bleeding. Thus, endoscopic evaluation is required to determine the need for urgent intervention in patients with gastric cancer and tumor bleeding.

Denture flask fabrication using fused deposition modeling three-dimensional printing.

PURPOSE: This paper describes a method for making a customized denture flask using fused deposition modeling (FDM) by three-dimensional (3D) printing. We have proposed a new digital dental prosthesis using conventional dental base materials and artificial teeth. METHODS: Using the universal development system software, a denture-designed Standard Tessellation Language (STL) file and a denture flask STL file were superimposed, and the denture region was set as an empty space. After setting the offset value to 200µm between the denture base and teeth for artificial tooth positioning, the flask was created by FDM 3D printing. Conventional artificial teeth were inserted into the 3D-printed flask, and resin packing, finishing, and polishing were performed using the conventional method for fabricating the complete denture. CONCLUSIONS: The 3D printing materials used to make digital dental prostheses have not yet been fully validated. Therefore, the production of a 3D-printed denture flask, which can use conventional complete denture materials, presents a new alternative to the digital fabrication of dentures.

Multi-omic biomarker identification and validation for diagnosing warzone-related post-traumatic stress disorder.

Post-traumatic stress disorder (PTSD) impacts many veterans and active duty soldiers, but diagnosis can be problematic due to biases in self-disclosure of symptoms, stigma within military populations, and limitations identifying those at risk. Prior studies suggest that PTSD may be a systemic illness, affecting not just the brain, but the entire body. Therefore, disease signals likely span multiple biological domains, including genes, proteins, cells, tissues, and organism-level physiological changes. Identification of these signals could aid in diagnostics, treatment decision-making, and risk evaluation. In the search for PTSD diagnostic biomarkers, we ascertained over one million molecular, cellular, physiological, and clinical features from three cohorts of male veterans. In a discovery cohort of 83 warzone-related PTSD cases and 82 warzone-exposed controls, we identified a set of 343 candidate biomarkers. These candidate biomarkers were selected from an integrated approach using (1) data-driven methods, including Support Vector Machine with Recursive Feature Elimination and other standard or published methodologies, and (2) hypothesis-driven approaches, using previous genetic studies for polygenic risk, or other PTSD-related literature. After reassessment of ~30% of these participants, we refined this set of markers from 343 to 28, based on their performance and ability to track changes in phenotype over time. The final diagnostic panel of 28 features was validated in an independent cohort (26 cases, 26 controls) with good performance (AUC = 0.80, 81% accuracy, 85% sensitivity, and 77% specificity). The identification and validation of this diverse diagnostic panel represents a powerful and novel approach to improve accuracy and reduce bias in diagnosing combat-related PTSD.