Inhibition of Glial Activation and Subsequent Reduction in White Matter Damage through Supplementation with a Combined Extract of Wheat Bran, Citrus Peel, and Jujube in a Rat Model of Vascular Dementia.

Vascular dementia (VaD) is the second most common type of dementia after Alzheimer's disease. In our previous studies, we showed that wheat bran extract (WBE) reduced white matter damage in a rat VaD model and improved memory in a human clinical trial. However, starch gelatinization made the large-scale preparation of WBE difficult. To simplify the manufacturing process and increase efficacy, we attempted to find a decoction containing an optimum ratio of wheat bran, sliced citrus peel, and sliced jujube (WCJ). To find an optimal ratio, the cell survival of C6 (rat glioma) cultured under hypoxic conditions (1% O2) was measured, and apoptosis was assessed. To confirm the efficacies of the optimized WCJ for VaD, pupillary light reflex, white matter damage, and the activation of astrocytes and microglia were assessed in a rat model of bilateral common carotid artery occlusion (BCCAO) causing chronic hypoperfusion. Using a combination of both searching the literature and cell survival experiments, we chose 6:2:1 as the optimal ratio of wheat bran to sliced citrus peel to sliced jujube to prepare WCJ. We showed that phytic acid contained only in wheat bran can be used as an indicator component for the quality control of WCJ. We observed in vitro that the WCJ treatment improved cell survival by reducing apoptosis through an increase in the Bcl-2/Bax ratio. In the BCCAO experiments, the WCJ-supplemented diet prevented astrocytic and microglial activation, mitigated myelin damage in the corpus callosum and optic tract, and, consequently, improved pupillary light reflex at dosages over 100 mg/kg/day. The results suggest that the consumption of WCJ can prevent VaD by reducing white matter damage, and WCJ can be developed as a safe, herbal medicine to prevent VaD.

Association Between Ambulance Station Case Volume and Clinical Outcomes in Moderate to Severe Trauma.

OBJECTIVES: The effect of the case volume of emergency medical services (EMS) on the clinical outcomes of trauma is uncertain. The purpose of this study was to evaluate the association between the case volume of an ambulance station and clinical outcomes in moderate to severe trauma patients. METHODS: Adult trauma patients with injury severity scores greater than 8 who were transported by the EMS between 2018 and 2019 were analyzed. The main exposure was the annual case volume of moderate to severe trauma at the ambulance station where the patient-transporting ambulance was based: low-volume (less than 60 cases), intermediate-volume (between 60 and 89 cases), and high-volume (equal or greater than 90 cases). The primary outcome was in-hospital mortality. Multilevel multivariable logistic regression analysis was conducted to calculate adjusted odds ratios (AORs) and 95% confidence intervals (CIs), with the high-volume group used as the reference. RESULTS: In total, 21,498 trauma patients were analyzed. The high-volume group exhibited lower in-hospital mortality, 447 (9.0%), compared to 867 (14.1%) in the intermediate-volume group and 1,458 (14.1%) in the low-volume group. There were a significantly higher odds of in-hospital mortality: the low-volume group (AOR 95% CI: 1.20 (0.95-1.51)) and intermediate-volume group (AOR 95% CI: 1.29 (1.02-1.64)) when compared to the high-volume group. CONCLUSIONS: The case volume at an ambulance station is associated with in-hospital mortality in patients with moderate to severe trauma. These results should be considered when constructing an EMS system and education program for prehospital trauma care.

Early identification of suspected serious infection among patients afebrile at initial presentation using neural network models and natural language processing: A development and external validation study in the emergency department.

OBJECTIVE: To develop and externally validate models based on neural networks and natural language processing (NLP) to identify suspected serious infections in emergency department (ED) patients afebrile at initial presentation. METHODS: This retrospective study included adults who visited the ED afebrile at initial presentation. We developed four models based on artificial neural networks to identify suspected serious infection. Patient demographics, vital signs, laboratory test results and information extracted from initial ED physician notes using term frequency-inverse document frequency were used as model variables. Models were trained and internally validated with data from one hospital and externally validated using data from a different hospital. Model discrimination was evaluated using area under the receiver operating characteristic curve (AUC) and 95% confidence intervals (CIs). RESULTS: The training, internal validation, and external validation datasets comprised 150,699, 37,675, and 85,098 patients, respectively. The AUCs (95% CIs) for Models 1 (demographics + vital signs), 2 (demographics + vital signs + initial ED physician note), 3 (demographics + vital signs + laboratory tests), and 4 (demographics + vital signs + laboratory tests + initial ED physician note) in the internal validation dataset were 0.789 (0.782-0.796), 0.867 (0.862-0.872), 0.881 (0.876-0.887), and 0.911 (0.906-0.915), respectively. In the external validation dataset, the AUCs (95% CIs) of Models 1, 2, 3, and 4 were 0.824 (0.817-0.830), 0.895 (0.890-0.899), 0.879 (0.873-0.884), and 0.913 (0.909-0.917), respectively. Model 1 can be utilized immediately after ED triage, Model 2 can be utilized after the initial physician notes are recorded (median time from ED triage: 28 min), and Models 3 and 4 can be utilized after the initial laboratory tests are reported (median time from ED triage: 68 min). CONCLUSIONS: We developed and validated models to identify suspected serious infection in the ED. Extracted information from initial ED physician notes using NLP contributed to increased model performance, permitting identification of suspected serious infection at early stages of ED visits.

Effect of no-flow period on the vasopressor effect of initial epinephrine administration in cardiac arrest.

OBJECTIVES: Whether a longer no-flow (NF) interval affects the magnitude of response to epinephrine in the resuscitation has not been well studied. Therefore, this study aimed to evaluate the effect of NF interval on the vasopressor effect of initial epinephrine administration in a porcine model. METHODS: We enrolled 20 pigs from two randomized porcine experimental studies using a ventricular fibrillation (VF) cardiac arrest model. The first experiment subjects were resuscitated after 4 min of NF (Short NF group), followed by three cycles (6 min) of chest compression using a mechanical cardiopulmonary resuscitation device before epinephrine administration. Second experiment subjects received 6 min of NF (Long NF group), two cycles (4 min) of chest compressions, and administration of epinephrine. Defibrillation for VF was delivered 8 and 10 min after VF induction in the Short NF and Long NF groups, respectively. The mean arterial pressure (MAP) and cerebral perfusion pressure (CePP) in the 2-min resuscitation period after epinephrine administration were compared between the study groups using the Wilcoxon rank-sum test. The mean differences in the parameters between phases were also compared. RESULTS: Seven pigs in the Short NF group and 13 pigs in the Long NF group were included in the analysis. All 2-min resuscitation phases from 6 to 16 min after VF induction were compared between the study groups. The Short NF group showed higher MAP and CePP in all phases (p < 0.01). Change of mean MAP after the epinephrine administration was significantly different between the study groups: mean difference (95% confidence interval) of 16.6 (15.8-17.4) mmHg in the Short NF group and 4.2 (3.9-4.5) mmHg in the Long NF group. CONCLUSION: In the porcine VF cardiac arrest model, 6 min of NF before resuscitation may affect the vasopressor effect of the initial epinephrine administered compared to 4 min of NF. A short NF may play a role in maximizing the effect of epinephrine in advanced cardiovascular life support.

Naturally occurring substitution in one amino acid in VHSV phosphoprotein enhances viral virulence in flounder.

Viral hemorrhagic septicemia virus (VHSV) is a rhabdovirus that causes high mortality in cultured flounder. Naturally occurring VHSV strains vary greatly in virulence. Until now, little has been known about genetic alterations that affect the virulence of VHSV in flounder. We recently reported the full-genome sequences of 18 VHSV strains. In this study, we determined the virulence of these 18 VHSV strains in flounder and then the assessed relationships between differences in the amino acid sequences of the 18 VHSV strains and their virulence to flounder. We identified one amino acid substitution in the phosphoprotein (P) (Pro55-to-Leu substitution in the P protein; PP55L) that is specific to highly virulent strains. This PP55L substitution was maintained stably after 30 cell passages. To investigate the effects of the PP55L substitution on VHSV virulence in flounder, we generated a recombinant VHSV carrying PP55L (rVHSV-P) from rVHSV carrying P55 in the P protein (rVHSV-wild). The rVHSV-P produced high level of viral RNA in cells and showed increased growth in cultured cells and virulence in flounder compared to the rVHSV-wild. In addition, rVHSV-P significantly inhibited the induction of the IFN1 gene in both cells and fish at 6 h post-infection. An RNA-seq analysis confirmed that rVHSV-P infection blocked the induction of several IFN-related genes in virus-infected cells at 6 h post-infection compared to rVHSV-wild. Ectopic expression of PP55L protein resulted in a decrease in IFN induction and an increase in viral RNA synthesis in rVHSV-wild-infected cells. Taken together, our results are the first to identify that the P55L substitution in the P protein enhances VHSV virulence in flounder. The data from this study add to the knowledge of VHSV virulence in flounder and could benefit VHSV surveillance efforts and the generation of a VHSV vaccine.

Effect of CRISPR/Cas9-mediated knockout of either IRF-3 or IRF-5 gene in Epithelioma papulosum cyprini cells on type I interferon response and NF-κB activity.

Transcription factors related to the activation of type I interferons (IFNs) and nuclear factor-kappa B (NF-κB) are known to be critical in innate immune responses. Interferon regulatory factors (IRFs) are a family of transcription factors. IRF-3 is known to act as the primary regulator in type I IFN signaling in response to viral infections, and the upregulation of IRF5 by virus infection has been reported in various fish species. One of the ways to know the functional role of certain genes is the production of target gene(s) knockout cells or organisms. In the present study, we produced either IRF3 or IRF5 gene knockout Epithelioma papulosum cyprini (EPC) cells using a CRISPR/Cas9 system, and investigated the effect of IRF3 gene and IRF5 gene knockout on polyinosinic:polycytidylic acid (ploly (I:C))-mediated and viral hemorrhagic septicemia virus (VHSV) infection-mediated type I IFN response and NF-κB activation. Both IRF3 knockout and IRF5 knockout EPC cells showed severely decreased type I IFN responses measured by ISRE activity and the expression of Mx1 and ISG15 genes when stimulated with poly (I:C), while the decreased level of type I IFN responses was not high as by poly (I:C) stimulation when infected with VHSV. Different from type I IFN response, NF-κB activities in IRF3 and IRF5 knockout cells were not highly different between poly (I:C) stimulated cells and VHSV-infected cells. Further studies are needed to elucidate pathways responsible for the type I IFN responses and NF-κB activation by VHSV infection.

Effect of temperature and IRF-9 gene-knockout on dynamics of vRNA, cRNA, and mRNA of viral hemorrhagic septicemia virus (VHSV).

The replication of viral hemorrhagic septicemia virus (VHSV) in appropriate host cells depends on environmental factors and the host cell's immunity. The dynamics of each VHSV RNA strand (vRNA, cRNA, and mRNA) in different conditions can provide a clue on the viral replication strategies, which can be a base for the development of efficient control measures. As VHSV is known to be sensitive to temperature and type I interferon (IFN) responses, in this study, we analyzed the effect of temperature difference (15 °C and 20 °C) and IRF-9 gene knockout on the dynamics of the three VHSV RNA strands in Epithelioma papulosum cyprini (EPC) cells using a strand-specific RT-qPCR. The tagged primers designed in this study successfully worked to quantify the three strands of VHSV. In the results of the temperature effect, the higher speed in viral mRNA transcription and the significantly higher (more than 10 times at 12-36 h) copy number of cRNA at 20 °C compared to those at 15 °C suggested the positive effect of high temperature on VHSV replication. In the results of the IRF-9 gene knockout effect, although IRF-9 gene knockout did not bring a dramatic effect on VHSV replication compared to the temperature effect, the increase of mRNA in IRF-9 KO cells was faster than normal EPC cells, which was reflected in the copy numbers of cRNA and vRNA. The IRF-9 gene knockout effect was not dramatic even in the replication of rVHSV-ΔNV-eGFP that harbors eGFP gene ORF instead of NV gene ORF. These results suggest that VHSV may be highly susceptible to pre-activated type I IFN responses but not highly susceptible to post-infection-mediated type I IFN responses or lowered type I IFN before infection. In both experiments of temperature effect and IRF-9 gene knockout effect, the copy number of cRNA never exceeded the copy number of vRNA at all assay times, suggesting that the binding efficiency of the RNP complex to the 3' end of cRNA might be lower than that to the 3' end of vRNA. Further research is needed to elucidate the regulatory mechanism that limits the amount of cRNA at an appropriate level during VHSV replication.

Protection of rainbow trout (Oncorhynchus mykiss) against VHSV genotype Ia and IHNV by immunization with VHSV genotype IVa backbone-based single-cycle viruses.

To evaluate the protective effect of viral hemorrhagic septicemia virus genotype IVa (VHSV IVa) genome-based single-cycle viruses against VHSV genotype Ia (VHSV Ia) and infectious hematopoietic necrosis virus (IHNV) in rainbow trout, three kinds of single-cycle VHSVs were rescued using reverse genetic technology: i) rVHSV-IaGΔTM-IVaG containing the transmembrane and cytoplasmic region-deleted G protein (GΔTM) of VHSV Ia instead of VHSV IVa full G gene ORF and having VHSV IVa G proteins on the envelope; ii) rVHSV-IaGΔTM-IaG containing VHSV Ia GΔTM instead of VHSV IVa full G gene ORF and having VHSV Ia G proteins on the envelope; iii) rVHSV-IaGΔTM-ihnvGΔTM-IVaG containing not only VHSV Ia GΔTM instead of full G gene but also IHNV GΔTM instead of NV gene and having VHSV IVa G proteins on the envelope. Rainbow trout immunized with rVHSV-IaGΔTM-IaG and rVHSV-IaGΔTM-IVaG showed significantly higher serum antibody titers against both VHSV Ia and VHSV IVa, and showed no mortality against VHSV Ia infection, while fish in the control groups showed 100% mortalities. Fish immunized with rVHSV-IaGΔTM-ihnvGΔTM-IVaG showed significantly higher serum antibody titers against VHSV IVa, VHSV Ia, and IHNV compared to fish in the control group. Immunization with rVHSV-IaGΔTM-ihnvGΔTM-IVaG induced significantly higher protection against not only VHSV Ia but also IHNV. These results suggest that the present single-cycle rVHSV-based system can be used as a platform to produce combined vaccines that can protect fish from multiple pathogenic species. However, the mechanism of the high protection against IHNV despite comparatively low antibody titer remains to be investigated.

Sex Disparities in Prehospital Advanced Cardiac Life Support in Out-of-Hospital Cardiac Arrest in South Korea.

OBJECTIVE: Sex disparities have been reported in the prehospital and in-hospital care among patients with out-of-hospital cardiac arrest (OHCA). The aim of this study was to investigate the association between sex and prehospital advanced cardiac life support (ACLS) interventions provided by emergency medical services (EMS). METHODS: This was a cross-sectional observational study using a nationwide OHCA registry in South Korea. The study included adult OHCAs with presumed cardiac etiology from January 2016 to December 2019. The main exposure was the sex of the victim, and the primary outcomes were prehospital ACLS interventions, including advanced airway management (AAM), intravenous access (IV), and epinephrine (EPI) administration. Multivariable logistic regression analysis accounted for age group, health insurance, comorbidities, place of arrest, urbanization level, witness status, bystander CPR and initial rhythm was performed to calculate adjusted odds ratios (AORs) with 95% confidence intervals (95% CIs). RESULTS: Among 71,154 eligible patients, females with OHCA received less prehospital ACLS interventions than males: risk difference, (95% CIs) -2.76 (-3.41;-2.11) for AAM, -6.03 (-6.79;-5.27) for IV, and -3.81 (-4.37;-3.25) for EPI. In multivariable logistic regression analysis, female sex was significantly associated with a lower probability of prehospital ACLS provision: AOR, (95% CIs) 0.87 (0.84-0.91) for AAM, 0.85 (0.82-0.88) for IV, and 0.81 (0.77-0.84) for EPI. CONCLUSION: Compared to male patients, female patients were less likely to receive prehospital ACLS. This offers opportunities for EMS systems to reduce disparities and to improve compliance with OHCA resuscitation guidelines and outcomes through quality improvement and educational interventions.

Association between prehospital airway type and oxygenation and ventilation in out-of-hospital cardiac arrest.

OBJECTIVES: This study aimed to evaluate the association between prehospital airway type and oxygenation and ventilation in out-of-hospital cardiac arrest (OHCA). METHODS: This retrospective observational study included OHCA patients who visited the emergency departments (EDs) between October 2015 and June 2021. The study groups were categorized according to the prehospital airway type: endotracheal intubation (ETI), supraglottic airway (SGA), or bag-valve-mask ventilation (BVM). The primary outcome was good oxygenation: partial pressure of oxygen (PaO2) ≥ 60 mmHg on the first arterial blood gas (ABG) test. The secondary outcome was good ventilation: partial pressure of carbon dioxide (PaCO2) ≤ 45 mmHg. Multivariate logistic regression was conducted to calculate the adjusted odds ratio (AOR) and 95% confidence interval (CI). RESULTS: A total of 7,372 patients were enrolled during the study period: 1,819 patients treated with BVM, 706 with ETI, and 4,847 who underwent SGA. In multivariable logistic regression analysis for good oxygenation outcomes, the ETI group showed a higher AOR than the BVM group (AOR [95% CIs]: 1.30 [1.06-1.59] in ETI and 1.05 [0.93-1.20] in SGA groups). Regarding good ventilation, the ETI group showed a higher AOR, and the SGA group showed a lower AOR compared to the BVM group (AOR [95% CIs] 1.33 [1.02-1.74] in the ETI and 0.83 (0.70-0.99) in the SGA groups). There was no significant difference in survival to discharge. CONCLUSIONS: ETI was significantly associated with good oxygenation and good ventilation compared to BVM in patients with OHCA, particularly during longer transports. This should be taken into consideration when deciding the prehospital advanced airway management in patients with OHCA.

A simple scoring rule to predict survival to discharge after out of hospital cardiac arrest at the time of ED arrival.

BACKGROUND: It is important to be able to predict the chance of survival to hospital discharge upon ED arrival in order to determine whether to continue or terminate resuscitation efforts after out of hospital cardiac arrest. This study was conducted to develop and validate a simple scoring rule that could predict survival to hospital discharge at the time of ED arrival. METHODS: This was a multicenter retrospective cohort study based on a nationwide registry (Korean Cardiac Arrest Research Consortium) of out of hospital cardiac arrest (OHCA). The study included adult OHCA patients older than 18 years old, who visited one of 33 tertiary hospitals in South Korea from September 1st, 2015 to June 30th, 2020. Among 12,321 screened, 5471 patients were deemed suitable for analysis after exclusion. Pre-hospital ROSC, pre-hospital witness, shockable rhythm, initial pH, and age were selected as the independent variables. The dependent variable was set to be the survival to hospital discharge. Multivariable logistic regression (LR) was performed, and the beta-coefficients were rounded to the nearest integer to formulate the scoring rule. Several machine learning algorithms including the random forest classifier (RF), support vector machine (SVM), and K-nearest neighbor classifier (K-NN) were also trained via 5-fold cross-validation over a pre-specified grid, and validated on the test data. The prediction performances and the calibration curves of each model were obtained. Pre-processing of the registry was done using R, model training & optimization using Python. RESULTS: A total of 5471 patients were included in the analysis. The AUROC of the scoring rule over the test data was 0.7620 (0.7311-0.7929). The AUROCs of the machine learning classifiers (LR, SVM, k-NN, RF) were 0.8126 (0.7748-0.8505), 0.7920 (0.7512-0.8329), 0.6783 (0.6236-0.7329), and 0.7879 (0.7465-0.8294), respectively. CONCLUSION: A simple scoring rule consisting of five, binary variables could aid in the prediction of the survival to hospital discharge at the time of ED arrival, showing comparable results to conventional machine learning classifiers.

Prehospital National Early Warning Score as a predictor of massive transfusion in adult trauma patients.

BACKGROUND: Previous studies have shown that an elevated prehospital National Early Warning Score (preNEWS) is associated with increased levels of adverse outcomes in patients with trauma. However, whether preNEWS is a predictor of massive transfusion (MT) in patients with trauma is currently unknown. This study investigated the accuracy of preNEWS in predicting MT and hospital mortality among trauma patients. METHODS: We analyzed adult trauma patients who were treated and transported by emergency medical services (EMS) between January 2018 and December 2019. The main exposure was the preNEWS calculated for the scene. The primary outcome was the predictive ability for MT, and the secondary outcome was 24 h mortality. We compared the prognostic performance of preNEWS with the shock index, modified shock index, and reverse shock index, and reverse shock index multiplied by Glasgow Coma Scale in the prehospital setting. RESULTS: In total, 41,852 patients were included, and 1456 (3.5%) received MT. preNEWS showed the highest area under the receiver operating characteristic (AUROC) curve for predicting MT (0.8504; 95% confidence interval [CI], 0.840-0.860) and 24 h mortality (AUROC 0.873; 95% CI, 0.863-0.883). The sensitivity of preNEWS for MT was 0.755, and the specificity of preNEWS for MT was 0.793. All indicies had a high negative predictive value and low positive predictive value. CONCLUSION: preNEWS is a useful, rapid predictor for MT and 24 h mortality. Calculation of preNEWS would be helpful for making the decision at the scene such as transfer straightforward to trauma center and advanced treatment.

Measurement of level of consciousness by AVPU scale assessment system based on automated video and speech recognition technology.

OBJECTIVE: To develop an alert/verbal/painful/unresponsive (AVPU) scale assessment system based on automated video and speech recognition technology (AVPU-AVSR) that can automatically assess a patient's level of consciousness and evaluate its performance through clinical simulation. METHODS: We developed an AVPU-AVSR system with a whole-body camera, face camera, and microphone. The AVPU-AVSR system automatically extracted essential audiovisual features to assess the AVPU score from the recorded video files. Arm movement, pain stimulus, and eyes-open state were extracted using a rule-based approach using landmarks estimated from pre-trained pose and face estimation models. Verbal stimuli were extracted using a pre-trained speech-recognition model. Simulations of a physician examining the consciousness of 12 simulated patients for 16 simulation scenarios (4 for each of "Alert", "Verbal", "Painful", and "Unresponsive") were conducted under the AVPU-AVSR system. The accuracy, sensitivity, and specificity of the AVPU-AVSR system were assessed. RESULTS: A total of 192 cases with 12 simulated patients were assessed using the AVPU-AVSR system with a multi-class accuracy of 0.95 (95% confidence interval [CI] (0.92-0.98). The sensitivity and specificity (95% CIs) for detecting impaired consciousness were 1.00 (0.97-1.00) and 0.88 (0.75-0.95), respectively. The sensitivity and specificity of each extracted feature ranged from 0.88 to 1.00 and 0.98 to 1.00. CONCLUSIONS: The AVPU-AVSR system showed good accuracy in assessing consciousness levels in a clinical simulation and has the potential to be implemented in clinical practice to automatically assess mental status.

A new genotype of decapod hepanhamaparvovirus 1 (DHPV) from cultured Penaeus vannamei in South Korea.

Decapod hepanhamaparvovirus 1 (DHPV), also known as hepatopancreatic parvovirus (HPV), has caused death in larvae or stunted growth in juveniles of cultured shrimp. To date, 4 genotypes (genotype I, II, III, and IV) have been reported from various shrimp species and various geographical regions. In the present study, we isolated 2 types of DHPV (GHPV-Goseong and DHPV-Geoje) from cultured Penaeus vannamei in Korea. Based on the capsid protein (VP) amino acid sequences, DHPV-Goseong was highly identical to previously reported DHPV genotype IV in Taiwan and Korea. Different from DHPV-Goseong, DHPV-Geoje showed approximately 63% similarity with DHPV genotype I, II, III and 84% similarity with DHPV genotype IV, suggesting an independent new genotype of DHPV (genotype V). Further research is needed to elucidate the origin and biological meanings of the present new genotype.

Capacitive Electrode-Based Electric Field Treatments on Redox-Toxic Iron Deposits in Transgenic AD Mouse Models: The Electroceutical Targeting of Alzheimer's Disease Feasibility Study.

Iron accumulation in the brain accelerates Alzheimer's disease progression. To cure iron toxicity, we assessed the therapeutic effects of noncontact transcranial electric field stimulation to the brain on toxic iron deposits in either the Aß fibril structure or the Aß plaque in a mouse model of Alzheimer's disease (AD) as a pilot study. A capacitive electrode-based alternating electric field (AEF) was applied to a suspension of magnetite (Fe3O4) to measure field-sensitized reactive oxygen species (ROS) generation. The increase in ROS generation compared to the untreated control was both exposure-time and AEF-frequency dependent. The frequency-specific exposure of AEF to 0.7-1.4 V/cm on a magnetite-bound Aß-fibril or a transgenic Alzheimer's disease (AD) mouse model revealed the degradation of the Aß fibril or the removal of the Aß-plaque burden and ferrous magnetite compared to the untreated control. The results of the behavioral tests show an improvement in impaired cognitive function following AEF treatment on the AD mouse model. Tissue clearing and 3D-imaging analysis revealed no induced damage to the neuronal structures of normal brain tissue following AEF treatment. In conclusion, our results suggest that the effective degradation of magnetite-bound amyloid fibrils or plaques in the AD brain by the electro-Fenton effect from electric field-sensitized magnetite offers a potential electroceutical treatment option for AD.

CRISPR/Cas9-mediated generation of auxotrophic Edwardsiella piscicida mutants and immunization in olive flounder (Paralichthys olivaceus).

Edwardsiella piscicida has been a cause of mass mortality in cultured fish. In this study, to produce auxotrophic E. piscicida mutants, a CRISPR/Cas9 system was used instead of the traditional sacB-based allelic exchange method. Under the optimal CRISPR engineering condition, we could efficiently produce either alr or asd gene knockout E. piscicida auxotrophic mutants, and this genome editing process was much simpler and faster than the allelic exchange method. The simultaneous knockout of double auxotrophic genes (alr and asd) and the insertion of a foreign gene expression cassette in E. piscicida chromosome were also successfully performed using the established CRISPR/Cas9 system. Furthermore, to enhance the possibility to get permission as a commercial vaccine, we produced an auxotrophic E. piscicida mutant having only one nucleotide-deleted alr gene (E. piscicida △alr-1). Olive flounder (Paralichthys olivaceus) fingerlings immunized with 1 × 106 and 1 × 105 CFU/fish of E. piscicida △alr-1 showed the superior ability in the induction of serum agglutination activity and in the protection against E. piscicida compared to killed E. piscicida. However, olive flounder immunized with 1 × 107 CFU/fish of E. piscicida △alr-1 showed high mortality far before the challenge, and the isolated E. piscicida from moribund and dead fish had the wild type alr gene, suggesting the reversion of one base-deleted alr gene to original form by a second mutation in olive flounder. Therefore, investigation on the minimum number of edited nucleotide for stable maintenance of E. piscicida mutants should be further conducted.

Effect of NV gene deletion in the genome of hirame rhabdovirus (HIRRV) on viral replication and the type I interferon response of the host cell.

Hirame rhabdovirus (HIRRV), a member of the genus Novirhabdovirus, causes morbidity and mortality in farmed olive flounder (Paralichthys olivaceus). As no information is available on the role of the NV gene of HIRRV, we produced a recombinant HIRRV with the NV gene deleted (rHIRRV-ΔNV) using reverse genetic technology and investigated whether the NV gene knockout affected HIRRV replication and the type I interferon response of the host cell. The rescue of rHIRRV-ΔNV was successful only when IRF9-gene-knockout Epithelioma papulosum cyprini (ΔIRF9-EPC) cells were used, suggesting that the NV protein of HIRRV might be involved in inhibition of the type I interferon response of the host cell. This conclusion was also supported by the significantly higher level of Mx gene induction in EPC cells infected with rHIRRV-ΔNV than in cells infected with recombinant HIRRV without the deletion. When cells were coinfected with rHIRRV-ΔNV and either wild-type HIRRV or wild-type viral hemorrhagic septicemia virus (VHSV), there was a decrease in the growth rate of not only wild-type HIRRV but also wild-type VHSV in a concentration-dependent manner. Further studies are required to investigate the role of HIRRV NV in virulence and its possible importance for the development of attenuated vaccines.

Association between Scene Time Interval and Survival in EMS-Treated Major Trauma Admitted to the Intensive Care Unit: A Multinational, Multicenter Observational Study.

Objective: Major trauma is a major concern in public health and a leading cause of mortality worldwide. This study aimed to evaluate the association between the prehospital scene time interval (STI) and survival in emergency medical service (EMS)-assessed major trauma patients admitted to the intensive care unit (ICU). Methods: A retrospective observational study using the Pan-Asian Trauma Outcomes Study (PATOS) database was conducted. Adult trauma patients with injury severity scores (ISSs) greater than 15 who were admitted to the ICU were selected. EMS STIs were categorized into three groups: short (0-8 minutes), intermediate (9-16 minutes), and long (over 16 minutes). The primary outcome was survival to hospital discharge, and the secondary outcome was good neurological outcome at hospital discharge. Multivariable logistic regression analysis was conducted to calculate odds ratios and confidence intervals, adjusting for age, sex, mechanism of injury, prehospital alertness, prehospital shock index, response time interval, and EMS intervention (airway, oxygen supplementation, and intravenous fluid administration). Sensitivity analysis for patients who underwent surgery or nontraumatic brain injury cases and interaction analysis by EMS intervention were performed. Results: Data from a total of 1,874 eligible patients were analyzed. Intermediate and long STIs showed significant associations with outcomes, with adjusted ORs (95% CI) of 1.21 (1.07-1.38) in the intermediate STI group and 1.74 (1.55-1.96) in the long STI group for survival and 1.37 (1.32-1.40) in the intermediate STI group and 1.31 (1.22-1.41) in the long STI group for neurological outcome. In the sensitivity analysis, the highest ORs were found in the intermediate STI group, with adjusted ORs (95% CI) of 1.40 (1.37-1.42) for survival and 1.32 (1.26-1.38) for neurological outcome. In the interaction analysis, EMS intervention showed a positive interaction effect with an intermediate STI on survival. Conclusion: In EMS-assessed adult major trauma patients admitted to the ICU, we found significant associations between STIs longer than 8 minutes and outcomes. EMS intervention has a positive interaction effect with an intermediate STI on survival. More research is needed to understand the implications of practice for major trauma in the field.

Prediction of bacteremia at the emergency department during triage and disposition stages using machine learning models.

INTRODUCTION: Bacteremia is a common but critical condition with high mortality that requires timely and optimal treatment in the emergency department (ED). The prediction of bacteremia at the ED during triage and disposition stages could support the clinical decisions of ED physicians regarding the appropriate treatment course and safe ED disposition. This study developed and validated machine learning models to predict bacteremia in the emergency department during triage and disposition stages. METHODS: This study enrolled adult patients who visited a single tertiary hospital from 2016 to 2018 and had at least two sets of blood cultures during their ED stay. Demographic information, chief complaint, triage level, vital signs, and laboratory data were used as model predictors. We developed and validated prediction models using 10 variables at the time of ED triage and 42 variables at the time of disposition. The extreme gradient boosting (XGB) model was compared with the random forest and multivariable logistic regression models. We compared model performance by assessing the area under the receiver operating characteristic curve (AUC), test characteristics, and decision curve analysis. RESULTS: A total of 24,768 patients were included: 16,197 cases were assigned to development, and 8571 cases were assigned to validation. The proportion of bacteremia was 10.9% and 10.4% in the development and validation datasets, respectively. The Triage XGB model (AUC, 0.718; 95% confidence interval (CI), 0.701-0.735) showed acceptable discrimination performance with a sensitivity over 97%. The Disposition XGB model (AUC, 0.853; 95% CI, 0.840-0.866) showed excellent performance and provided the greatest net benefit throughout the range of thresholds probabilities. CONCLUSIONS: The Triage XGB model could be used to identify patients with a low risk of bacteremia immediately after initial ED triage. The Disposition XGB model showed excellent discriminative performance.

Association between focused cardiac ultrasound and time to furosemide administration in acute heart failure.

BACKGROUND: Heart failure (HF) is a global health burden, and its management in the emergency department (ED) is important. This study aimed to evaluate the association between focused cardiac ultrasound (FoCUS) and early administration of diuretics in patients with acute HF admitted to the ED. METHODS: This retrospective observational study was conducted at a tertiary academic hospital. Patients with acute HF patients who were admitted to the ED and receiving intravenous medication between January 2018 and December 2019 were enrolled. The main exposure was a FoCUS examination performed within 2 h of ED triage. The primary outcome was the time to furosemide administration. RESULTS: Of 1154 patients with acute HF, 787 were included in the study, with 116 of them having undergone FoCUS. The time to furosemide was significantly shorter in the FoCUS group (median time (q1-q3), 112 min; range, 65-163 min) compared to the non-FoCUS group (median time, 131 min; range, 71-229 min). In the multivariable logistic regression analysis adjusting for age, sex, chief complaint, mode of arrival, triage level, shock status, and desaturation at triage, early administration of furosemide within 2 h from triage was significantly higher in the FoCUS group (adjusted odds ratio, 1.63; 95% confidence intervals, 1.04-2.55) than in the non-FoCUS group. CONCLUSIONS: Early administration of intravenous furosemide was associated with FoCUS examination in patients with acute HF admitted to the ED. An early screening protocol could be useful for improving levels in clinical practice at EDs.