CcpA and CodY Regulate CRISPR-Cas System of Streptococcus mutans.

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) genes are widely recognized as bacterial adaptive immune systems against invading viruses and bacteriophages. The oral pathogen Streptococcus mutans encodes two CRISPR-Cas loci (CRISPR1-Cas and CRISPR2-Cas), and their expression under environmental conditions is still under investigation. In this study, we investigated the transcriptional regulation of cas operons by CcpA and CodY, two global regulators that contribute to carbohydrate and (p)ppGpp metabolism. The possible promoter regions for cas operons and the binding sites for CcpA and CodY in the promoter regions of both CRISPR-Cas loci were predicted using computational algorithms. We found that CcpA could directly bind to the upstream region of both cas operons, and detected an allosteric interaction of CodY within the same region. The binding sequences of the two regulators were identified through footprinting analysis. Our results showed that the promoter activity of CRISPR1-Cas was enhanced under fructose-rich conditions, while deletion of the ccpA gene led to reduced activity of the CRISPR2-Cas promoter under the same conditions. Additionally, deletion of the CRISPR systems resulted in a significant decrease in fructose uptake ability compared to the parental strain. Interestingly, the accumulation of guanosine tetraphosphate (ppGpp) was reduced in the presence of mupirocin, which induces a stringent response, in the CRISPR1-Cas-deleted (ΔCR1cas) and both CRISPR-Cas-deleted (ΔCRDcas) mutant strains. Furthermore, the promoter activity of both CRISPRs was enhanced in response to oxidative or membrane stress, while the CRISPR1 promoter activity was reduced under low-pH conditions. Collectively, our findings demonstrate that the transcription of the CRISPR-Cas system is directly regulated by the binding of CcpA and CodY. These regulatory actions play a crucial role in modulating glycolytic processes and exerting effective CRISPR-mediated immunity in response to nutrient availability and environmental cues. IMPORTANCE An effective immune system has evolved not only in eukaryotic organisms but also in microorganisms, enabling them to rapidly detect and neutralize foreign invaders in the environment. Specifically, the CRISPR-Cas system in bacterial cells is established through a complex and sophisticated regulatory mechanism involving specific factors. In this study, we demonstrate that the expression of two CRISPR systems in S. mutans can be controlled by two global regulators, CcpA and CodY, which play critical roles in carbohydrate metabolism and amino acid biosynthesis. Importantly, our results show that the expression of the CRISPR-Cas system in S. mutans influences (p)ppGpp production during the stringent response, which is a gene expression regulatory response that aids in environmental stress adaptation. This transcriptional regulation by these regulators enables a CRISPR-mediated immune response in a host environment with limited availability of carbon sources or amino acids, while ensuring efficient carbon flux and energy expenditure to support multiple metabolic processes.

The Auxiliary Role of Virtual Reality in Enhancing the Effects of Disaster News on Empathy and Fear: The Mediating Role of Presence.

This study conducted a two-condition between-subjects laboratory experiment (N = 147) to examine the effects of textual disaster news combined with a 360° video in different modes (virtual reality [VR] vs. two dimensional screen) on cognitive/affective empathy and fear. Additionally, the mediating role of presence (social and spatial presence) was considered. Results indicated that the news article presented through an immersive VR mode directly led to greater levels of presence and cognitive empathy but not affective empathy and fear. The effects on social presence further resulted in greater cognitive and affective empathy, not fear. Meanwhile, spatial presence enhanced by VR led to greater levels of emotions (affective empathy and fear) but not cognitive empathy. These findings suggest the dynamic mechanisms of how an immersive VR modality can function as a complementary tool for traditional news to improve audience engagement, which centers on emotion-related outcomes in tandem with cognitive responses.

Asthma and increased risk of myocardial infarction and mortality among hypertensive Korean patients.

This study aimed to evaluate the effects of asthma on cardiovascular disease incidence in patients with hypertension. A total of 639,784 patients with hypertension from the Korea National Health Insurance Service database were included, of whom 62,517 had history of asthma after propensity score matching. The risks of all-cause mortality, myocardial infarction (MI), stroke, and end-stage renal disease (ESRD) were assessed according to the presence of asthma, long-acting ß2-agonist (LABA) inhaler usage, and/or systemic corticosteroid usage for up to 11 years. In addition, whether these risks were modified by average blood pressure (BP) levels during the follow-up period was examined. Asthma was associated with an increased risk of all-cause mortality (hazard ratio [HR], 1.203; 95% confidence interval [CI], 1.165-1.241) and MI (HR, 1.244; 95% CI, 1.182-1.310) but not the risk of stroke or ESRD. LABA inhaler usage was associated with a higher risk of all-cause mortality and MI, and systemic corticosteroids usage showed a higher risk of ESRD as well as all-cause mortality and MI among hypertensive patients with asthma. Compared to patients without asthma, there was a graded increase in the risk of all-cause mortality and MI in those with asthma without LABA inhaler/systemic corticosteroid usage and in those with asthma with LABA inhaler/systemic corticosteroid usage. These associations were not significantly modified by BP levels. This nationwide population-based study supports that asthma may be a clinical factor that increases the risk of poor outcomes in patients with hypertension.

Inverse Identification of a Constitutive Model for High-Speed Forming Simulation: An Application to Electromagnetic Metal Forming.

Forming simulation requires a constitutive model whose parameters are typically determined with tensile tests assumed static. However, this conventional approach is impractical for high-speed forming simulation characterized by high strain rates inducing transient effects. To identify constitutive parameters in relation to high-speed forming simulation, we formulated the problem of constitutive modeling as inverse parameter estimation addressed by regularized nonlinear least squares. Regarding the proposed inverse constitutive modeling, we adopted the L-curve method for proper regularization and model order reduction for rapid simulation. For demonstration, we corroborated the proposed strategy by identifying the modified Johnson-Cook model in the context of a free bulge test with electromagnetic metal forming simulation. The L-curve method allowed us to systematically choose a regularization parameter, and model order reduction brought enormous computational savings. After identifying constitutive parameters, we successfully verified and validated the reduced and original simulation models, respectively, with a manufactured workpiece. In addition, we validated the numerically identified constitutive model with a dynamic material test using a split Hopkinson pressure bar. Overall, we showed that inverse constitutive modeling for high-speed forming simulation can be effectively tackled by regularized nonlinear least squares with the help of an L-curve and a reduced-order model.

Remarkable enantioselectivity enhancement of the extractors with nonaxial chirality in liquid-liquid extraction of underivatized amino acids by introducing t-butyl group.

A class of carbonyl extractors, (R)-3, (R)-4, and (R)-5, with nonaxial chirality containing asymmetric carbons has been synthesized and studied for their efficiencies in enantioselective liquid-liquid extraction for underivatized amino acids. The bulky t-butyl ketone extractors, (R)-4 and (R)-5, showed the stereoselectivities ranging 5.4-9.4 of l/d ratio much better than those of the aldehyde extractor, (R)-3, ranging 2.4-5.2. The imine formation rates and yields of the t-butyl ketones were not significantly affected by their bulkiness and even in the absence of resonance-assisted hydrogen bond. This work confirms that a bulky t-butyl ketone can be a good choice in the development of an extractor not only with axial chirality but also with nonaxial chirality for the enantioselective extraction of unprotected amino acids.

Multifunctional Injectable Hydrogel for In Vivo Diagnostic and Therapeutic Applications.

Injectable hydrogels show high potential for in vivo biomedical applications owing to their distinctive mode of administration into the human body. In this study, we propose a material design strategy for developing a multifunctional injectable hydrogel with good adhesiveness, stretchability, and bioresorbability. Its multifunctionality, whereupon multiple reactions occur simultaneously during its injection into the body without requiring energy stimuli and/or additives, was realized through meticulous engineering of bioresorbable precursors based on hydrogel chemistry. The multifunctional injectable hydrogel can be administered through a minimally invasive procedure, form a conformal adhesive interface with the target tissue, dynamically stretch along with the organ motions with minimal mechanical constraints, and be resorbed in vivo after a specific period. Further, the incorporation of functional nanomaterials into the hydrogel allows for various in vivo diagnostic and therapeutic applications, without compromising the original multifunctionality of the hydrogel. These features are verified through theranostic case studies on representative organs, including the skin, liver, heart, and bladder.

Deep-learning model for screening sepsis using electrocardiography.

BACKGROUND: Sepsis is a life-threatening organ dysfunction and a major healthcare burden worldwide. Although sepsis is a medical emergency that requires immediate management, screening for the occurrence of sepsis is difficult. Herein, we propose a deep learning-based model (DLM) for screening sepsis using electrocardiography (ECG). METHODS: This retrospective cohort study included 46,017 patients who were admitted to two hospitals. A total of 1,548 and 639 patients had sepsis and septic shock, respectively. The DLM was developed using 73,727 ECGs from 18,142 patients, and internal validation was conducted using 7774 ECGs from 7,774 patients. Furthermore, we conducted an external validation with 20,101 ECGs from 20,101 patients from another hospital to verify the applicability of the DLM across centers. RESULTS: During the internal and external validations, the area under the receiver operating characteristic curve (AUC) of the DLM using 12-lead ECG was 0.901 (95% confidence interval, 0.882-0.920) and 0.863 (0.846-0.879), respectively, for screening sepsis and 0.906 (95% confidence interval (CI), 0.877-0.936) and 0.899 (95% CI, 0.872-0.925), respectively, for detecting septic shock. The AUC of the DLM for detecting sepsis using 6-lead and single-lead ECGs was 0.845-0.882. A sensitivity map revealed that the QRS complex and T waves were associated with sepsis. Subgroup analysis was conducted using ECGs from 4,609 patients who were admitted with an infectious disease, and the AUC of the DLM for predicting in-hospital mortality was 0.817 (0.793-0.840). There was a significant difference in the prediction score of DLM using ECG according to the presence of infection in the validation dataset (0.277 vs. 0.574, p < 0.001), including severe acute respiratory syndrome coronavirus 2 (0.260 vs. 0.725, p = 0.018). CONCLUSIONS: The DLM delivered reasonable performance for sepsis screening using 12-, 6-, and single-lead ECGs. The results suggest that sepsis can be screened using not only conventional ECG devices but also diverse life-type ECG machines employing the DLM, thereby preventing irreversible disease progression and mortality.

Performance Evaluation of the Newly Developed In Vitro Rapid Diagnostic Test for Detecting OXA-48-Like, KPC-, NDM-, VIM- and IMP-Type Carbapenemases: The RESIST-5 O.K.N.V.I. Multiplex Lateral Flow Assay.

The objective of this study was to evaluate the performance of the RESIST-5 O.K.N.V.I. assay for identifying these five common domestic carbapenemases among a large number of clinical isolates in South Korea. A total of 268 non-duplicated clinical isolates of gram-negative bacilli were included in this study as follows: 258 carbapenemase-producing (CP) strains (OXA-48-like, KPC, NDM, VIM, IMP, GES, OXA-23 and two or more carbapenemase producers) and 10 non-CP carbapenem-resistant Enterobacterales (non-CP CREs). Overall sensitivity and specificity were 98.4% and 100%, respectively. In addition, all non-targeted carbapenemase producers including GES and OXA-23 producers and non-CP CREs were correctly identified as negative results. There were only four discrepant cases in which three VIM carbapenemase producers and one NDM carbapenemase producer were not detected. The RESIST-5 O.K.N.V.I. assay as an in vitro diagnostic test for detecting five common carbapenemases provided rapid and accurate results in a short time, indicating that this method could provide an innovative solution for early detection, resulting in appropriate antimicrobial treatment in the clinical field.

A Biodegradable Secondary Battery and its Biodegradation Mechanism for Eco-Friendly Energy-Storage Systems.

The production of rechargeable batteries is rapidly expanding, and there are going to be new challenges in the near future about how the potential environmental impact caused by the disposal of the large volume of the used batteries can be minimized. Herein, a novel strategy is proposed to address these concerns by applying biodegradable device technology. An eco-friendly and biodegradable sodium-ion secondary battery (SIB) is developed through extensive material screening followed by the synthesis of biodegradable electrodes and their seamless assembly with an unconventional biodegradable separator, electrolyte, and package. Each battery component decomposes in nature into non-toxic compounds or elements via hydrolysis and/or fungal degradation, with all of the biodegradation products naturally abundant and eco-friendly. Detailed biodegradation mechanisms and toxicity influence of each component on living organisms are determined. In addition, this new SIB delivers performance comparable to that of conventional non-degradable SIBs. The strategy and findings suggest a novel eco-friendly biodegradable paradigm for large-scale rechargeable battery systems.

Enantioselective extraction of unprotected amino acids coupled with racemization.

Scalable and economical methods for the production of optically pure amino acids, both natural and unnatural, are essential for their use as synthetic building blocks. Currently, enzymatic dynamic kinetic resolution (DKR) underpins some of the most effective processes. Here we report the development of enantioselective extraction coupled with racemization (EECR) for the chirality conversion of underivatized amino acids. In this process, the catalytic racemization of amino acids in a basic aqueous solution is coupled with the selective extraction of one enantiomer into an organic layer. Back-extraction from the organic layer to an acidic aqueous solution then completes the deracemization of the amino acid. The automation of the EECR process in a recycling flow reactor is also demonstrated. Continuous EECR is made possible by the sterically hindered chiral ketone extractant 5, which prevents the coextraction of the copper racemization catalyst because of its nonplanar geometry. Furthermore, the extractant 5 unexpectedly forms imines with amino acids faster and with greater enantioselectivity than less bulky derivatives, even though 5 cannot participate in intramolecular resonance-assisted hydrogen bonding. These features may allow EECR to challenge the preponderance of enzymatic DKR in the production of enantiomerically enriched amino acids.

Phthalate Plasticizers in Children's Products and Estimation of Exposure: Importance of Migration Rate.

Plasticizers are added to diverse consumer products including children's products. Owing to their potential for endocrine disruption, the use of phthalate plasticizers is restricted in many children's products. In this study, exposure to five phthalate esters (dibutylphthalate, di(2-ethylhexyl) phthalate (DEHP), diethyl phthalate, di-isobutyl phthalate, and diisononyl phthalate (DINP)) and an alternative (di-ethylhexyl adipate) was assessed by the use of children's products based on chemical analysis of 3345 products purchased during 2017 and 2019 in Korea. Plasticizers were found above the detection limits in 387 products, and DEHP and DINP were the two most predominantly detected plasticizers. Deterministic and probabilistic estimation of the margin of exposure at a screening level revealed that the use of children's products might be an important risk factor. However, it is also highly likely that the exposure could be overestimated, because the migration rate was estimated based solely on the content of plasticizers in children's products. Chemical migration is a key process determining the absorption of plasticizers from products; thus, further refinements in experimental determination or model estimation of the migration rate are required.

Microplastics in Food: A Review on Analytical Methods and Challenges.

Human exposure to microplastics contained in food has become a significant concern owing to the increasing accumulation of microplastics in the environment. In this paper, we summarize the presence of microplastics in food and the analytical methods used for isolation and identification of microplastics. Although a large number of studies on seafood such as fish and shellfish exist, estimating the overall human exposure to microplastics via food consumption is difficult owing to the lack of studies on other food items. Analytical methods still need to be optimized for appropriate recovery of microplastics in various food matrices, rendering a quantitative comparison of different studies challenging. In addition, microplastics could be added or removed from ingredients during processing or cooking. Thus, research on processed food is crucial to estimate the contribution of food to overall human microplastic consumption and to mitigate this exposure in the future.

Artificial intelligence algorithm to predict the need for critical care in prehospital emergency medical services.

BACKGROUND: In emergency medical services (EMSs), accurately predicting the severity of a patient's medical condition is important for the early identification of those who are vulnerable and at high-risk. In this study, we developed and validated an artificial intelligence (AI) algorithm based on deep learning to predict the need for critical care during EMS. METHODS: We conducted a retrospective observation cohort study. The algorithm was established using development data from the Korean national emergency department information system, which were collected during visits in real time from 151 emergency departments (EDs). We validated the algorithm using EMS run sheets from two EDs. The study subjects comprised adult patients who visited EDs. The endpoint was critical care, and we used age, sex, chief complaint, symptom onset to arrival time, trauma, and initial vital signs as the predicted variables. RESULTS: The number of patients in the development data was 8,981,181, and the validation data comprised 2604 EMS run sheets from two hospitals. The area under the receiver operating characteristic curve of the algorithm to predict the critical care was 0.867 (95% confidence interval, [0.864-0.871]). This result outperformed the Emergency Severity Index (0.839 [0.831-0.846]), Korean Triage and Acuity System (0.824 [0.815-0.832]), National Early Warning Score (0.741 [0.734-0.748]), and Modified Early Warning Score (0.696 [0.691-0.699]). CONCLUSIONS: The AI algorithm accurately predicted the need for the critical care of patients using information during EMS and outperformed the conventional triage tools and early warning scores.

Flexible, sticky, and biodegradable wireless device for drug delivery to brain tumors.

Implantation of biodegradable wafers near the brain surgery site to deliver anti-cancer agents which target residual tumor cells by bypassing the blood-brain barrier has been a promising method for brain tumor treatment. However, further improvement in the prognosis is still necessary. We herein present novel materials and device technologies for drug delivery to brain tumors, i.e., a flexible, sticky, and biodegradable drug-loaded patch integrated with wireless electronics for controlled intracranial drug delivery through mild-thermic actuation. The flexible and bifacially-designed sticky/hydrophobic device allows conformal adhesion on the brain surgery site and provides spatially-controlled and temporarily-extended drug delivery to brain tumors while minimizing unintended drug leakage to the cerebrospinal fluid. Biodegradation of the entire device minimizes potential neurological side-effects. Application of the device to the mouse model confirms tumor volume suppression and improved survival rate. Demonstration in a large animal model (canine model) exhibited its potential for human application.

Handheld Microflow Cytometer Based on a Motorized Smart Pipette, a Microfluidic Cell Concentrator, and a Miniaturized Fluorescence Microscope.

Miniaturizing flow cytometry requires a comprehensive approach to redesigning the conventional fluidic and optical systems to have a small footprint and simple usage and to enable rapid cell analysis. Microfluidic methods have addressed some challenges in limiting the realization of microflow cytometry, but most microfluidics-based flow cytometry techniques still rely on bulky equipment (e.g., high-precision syringe pumps and bench-top microscopes). Here, we describe a comprehensive approach that achieves high-throughput white blood cell (WBC) counting in a portable and handheld manner, thereby allowing the complete miniaturization of flow cytometry. Our approach integrates three major components: a motorized smart pipette for accurate volume metering and controllable liquid pumping, a microfluidic cell concentrator for target cell enrichment, and a miniaturized fluorescence microscope for portable flow cytometric analysis. We first validated the capability of each component by precisely metering various fluid samples and controlling flow rates in a range from 219.5 to 840.5 µL/min, achieving high sample-volume reduction via on-chip WBC enrichment, and successfully counting single WBCs flowing through a region of interrogation. We synergistically combined the three major components to create a handheld, integrated microflow cytometer and operated it with a simple protocol of drawing up a blood sample via pipetting and injecting the sample into the microfluidic concentrator by powering the motorized smart pipette. We then demonstrated the utility of the microflow cytometer as a quality control means for leukoreduced blood products, quantitatively analyzing residual WBCs (rWBCs) in blood samples present at concentrations as low as 0.1 rWBCs/µL. These portable, controllable, high-throughput, and quantitative microflow cytometric technologies provide promising ways of miniaturizing flow cytometry.

Bioresorbable Electronic Implants: History, Materials, Fabrication, Devices, and Clinical Applications.

Medical implants, either passive implants for structural support or implantable devices with active electronics, have been widely used for the diagnosis and treatment of various diseases and clinical issues. These implants offer various functions, including mechanical support of biological structures in orthopedic and dental applications, continuous electrophysiological monitoring and feedback of electrical stimulation in neuronal and cardiac applications, and controlled drug delivery while maintaining arterial structure in drug-eluting stents. Although these implants exhibit long-term biocompatibility, surgery for their retrieval is often required, which imposes physical, biological, and economical burdens on the patients. Therefore, as an alternative to such secondary surgeries, bioresorbable implants that disappear after a certain period of time inside the body, including bioresorbable active electronics, have been highlighted recently. This review first discusses the historical background of medical implants and briefly define related terminology. Representative examples of non-degradable medical implants for passive structural support and/or for diagnosis and therapy with active electronics are also provided. Then, recent progress in bioresorbable active implants composed of biosignal sensors, actuators for therapeutics, wireless power supply components, and their integrated systems are reviewed. Finally, clinical applications of these bioresorbable electronic implants are exemplified with brief conclusion and future outlook.

Anti-Obesity Effects of a Mixture of Fermented Ginseng, Bifidobacterium longum BORI, and Lactobacillus paracasei CH88 in High-Fat Diet-Fed Mice.

Ginseng and probiotics have anti-obesity effects in mice fed a high-fat diet (HFD). Absorption of ginsenoside and colonization of probiotics occur in the intestine. In this study, a mixture of fermented ginseng and two probiotics, Bifidobacterium longum BORI and Lactobacillus paracasei CH88, was administered to HFD-fed mice for 9 weeks. The mixture significantly suppressed weight gain (p < 0.05, n = 8) and lipid deposition in the liver and adipose tissues as well as increased the mice's food intake. The adipocyte size of the adipose tissue was significantly decreased in the mixture-fed group, especially when 0.5% fermented ginseng and 5 × 108/ml of the two probiotics were used (p < 0.05, n = 10). The expression of TNF-α in adipose tissue was efficiently downregulated in the mixture-fed group (p < 0.05, n = 4). The supplement also improved the mice's fasting blood glucose levels (p < 0.05, n = 8) and total cholesterol feces excretion (p < 0.05, n = 8). The mixture of fermented ginseng and B. longum BORI and L. paracasei CH88 could have an anti-obesity effect and suppress lipid deposit in the liver and adipose tissues.

Is There a Difference in Stability After Intraoral Vertical Ramus Osteotomy Between Vertically High-Angle and Normal-Angle Patients?

PURPOSE: Few studies have evaluated the relapse pattern of intraoral vertical ramus osteotomy (IVRO) for the correction of mandibular prognathism with a high angle. The aim of this study was to measure the association between vertical facial types (high and normal mandibular plane angle) and relapse after IVRO for the management of mandibular prognathism. MATERIALS AND METHODS: The retrospective cohort study sample (skeletal Class III patients) was divided into 2 groups according to the angle of the sella-nasion plane relative to the mandibular plane (SN-MP) at the initial examination. Lateral cephalograms were analyzed for the predictor (facial type) and outcome (cephalometric changes over time) variables before surgery, 7 days after surgery, and 12 months after surgery. The 2 groups were matched for sample size (n = 20 in each). Data were analyzed using repeated-measures analysis of variance with Bonferroni correction. RESULTS: The normal-angle group (group N, SN-MP from 27° to 37°) and high-angle group (group H, SN-MP >37°) were not significantly different in terms of gender and age at the initial examination. Seven days after surgery, the mandibles in group H moved 2.5 mm more superiorly than those in group N (P = .013); consequently, the amount of overbite correction in group H was approximately 2 mm greater than that in group N (P = .002). Nevertheless, 12 months after surgery, there was no statistically significant difference in relapse of the maxilla and mandible between the 2 groups. In the 2 groups, the mandible moved approximately 0.7 mm superiorly during retention. CONCLUSIONS: These findings suggest that IVRO is a clinically acceptable and stable treatment modality for mandibular prognathism with a high angle.

Three-Dimensional Polymeric Mechanical Metamaterials Fabricated by Multibeam Interference Lithography with the Assistance of Plasma Etching.

The pentamode structure is a type of mechanical metamaterial that displays dramatically different bulk and shear modulus responses. In this study, a face-centered cubic (FCC) polymeric microstructure was fabricated by using SU8 negative-type photoresists and multibeam interference exposure. Isotropic plasma etching is used to control the solid-volume fraction; for the first time, we obtained a structure with the minimum solid-volume fraction as low as 15% that still exhibited high structural integrity. Using this method, we reduced the width of atom-to-atom connections by up to 40 nm. We characterize the effect of the connection area on the anisotropy of the mechanical properties using simulations. Nanoindentation measurements were also conducted to evaluate the energy dissipation by varying the connection area. The Young's/shear modulus ratio is 5 times higher for the etched microstructure than that of the bulk SU8 materials. The use of interference lithography may enable the properties of microscale materials to be engineered for various applications, such as MEMS.