GPCR targeting of E3 ubiquitin ligase MDM2 by inactive ß-arrestin.

E3 ubiquitin ligase Mdm2 facilitates ß-arrestin ubiquitination, leading to the internalization of G protein-coupled receptors (GPCRs). In this process, ß-arrestins bind to Mdm2 and recruit it to the receptor; however, the molecular architecture of the ß-arrestin-Mdm2 complex has not been elucidated yet. Here, we identified the ß-arrestin-binding region (ABR) on Mdm2 and solved the crystal structure of ß-arrestin1 in complex with Mdm2ABR peptide. The acidic residues of Mdm2ABR bind to the positively charged concave side of the ß-arrestin1 N-domain. The C-tail of ß-arrestin1 is still bound to the N-domain, indicating that Mdm2 binds to the inactive state of ß-arrestin1, whereas the phosphorylated C-terminal tail of GPCRs binds to activate ß-arrestins. The overlapped binding site of Mdm2 and GPCR C-tails on ß-arrestin1 suggests that the binding of GPCR C-tails might trigger the release of Mdm2. Moreover, hydrogen/deuterium exchange experiments further show that Mdm2ABR binding to ß-arrestin1 induces the interdomain interface to be more dynamic and uncouples the IP6-induced oligomer of ß-arrestin1. These results show how the E3 ligase, Mdm2, interacts with ß-arrestins to promote the internalization of GPCRs.

Cyclin Y regulates spatial learning and memory flexibility through distinct control of the actin pathway.

Spatial learning and memory flexibility are known to require long-term potentiation (LTP) and long-term depression (LTD), respectively, on a cellular basis. We previously showed that cyclin Y (CCNY), a synapse-remodeling cyclin, is a novel actin-binding protein and an inhibitory regulator of functional and structural LTP in vitro. In this study, we report that Ccny knockout (KO) mice exhibit enhanced LTP and weak LTD at Schaffer collateral-CA1 synapses in the hippocampus. In accordance with enhanced LTP, Ccny KO mice showed improved spatial learning and memory. However, although previous studies reported that normal LTD is necessary for memory flexibility, Ccny KO mice intriguingly showed improved memory flexibility, suggesting that weak LTD could exert memory flexibility when combined with enhanced LTP. At the molecular level, CCNY modulated spatial learning and memory flexibility by distinctively affecting the cofilin-actin signaling pathway in the hippocampus. Specifically, CCNY inhibited cofilin activation by original learning, but reversed such inhibition by reversal learning. Furthermore, viral-mediated overexpression of a phosphomimetic cofilin-S3E in hippocampal CA1 regions enhanced LTP, weakened LTD, and improved spatial learning and memory flexibility, thus mirroring the phenotype of Ccny KO mice. In contrast, the overexpression of a non-phosphorylatable cofilin-S3A in hippocampal CA1 regions of Ccny KO mice reversed the synaptic plasticity, spatial learning, and memory flexibility phenotypes observed in Ccny KO mice. Altogether, our findings demonstrate that LTP and LTD cooperatively regulate memory flexibility. Moreover, CCNY suppresses LTP while facilitating LTD in the hippocampus and negatively regulates spatial learning and memory flexibility through the control of cofilin-actin signaling, proposing CCNY as a learning regulator modulating both memorizing and forgetting processes.

One-Pot Three-Component Reaction for the Synthesis of 3,4-Dihydroquinazolines and Quinazolin-4(3H)-ones.

A highly efficient and straightforward one-pot synthesis of diversely substituted 3,4-dihydroquinazolines and quinazolin-4(3H)-ones has been achieved through a domino three-component assembly reaction of arenediazonium salts, nitriles, and bifunctional aniline derivatives. This new protocol involves three C-N bond formations through the initial formation of N-arylnitrilium intermediates from arenediazonium salts and nitriles, followed by the sequential nucleophilic addition and cyclization reactions with bifunctional anilines, leading to such N-heterocyclic compounds of biological and pharmacological importance. This method offers a simple, expedient, and robust approach with the use of amenable and easily accessible reactants/reagents under metal-free mild conditions, good functional group tolerance, and high efficiency. The synthetic applications were also demonstrated by derivatization of the products obtained from these processes and syntheses of a diverse range of valuable polycyclic N-heterocycles.

FOXL2 and FOXA1 cooperatively assemble on the TP53 promoter in alternative dimer configurations.

Although both the p53 and forkhead box (FOX) family proteins are key transcription factors associated with cancer progression, their direct relationship is unknown. Here, we found that FOX family proteins bind to the non-canonical homotypic cluster of the p53 promoter region (TP53). Analysis of crystal structures of FOX proteins (FOXL2 and FOXA1) bound to the p53 homotypic cluster indicated that they interact with a 2:1 stoichiometry accommodated by FOX-induced DNA allostery. In particular, FOX proteins exhibited distinct dimerization patterns in recognition of the same p53-DNA; dimer formation of FOXA1 involved protein-protein interaction, but FOXL2 did not. Biochemical and biological functional analyses confirmed the cooperative binding of FOX proteins to the TP53 promoter for the transcriptional activation of TP53. In addition, up-regulation of TP53 was necessary for FOX proteins to exhibit anti-proliferative activity in cancer cells. These analyses reveal the presence of a discrete characteristic within FOX family proteins in which FOX proteins regulate the transcription activity of the p53 tumor suppressor via cooperative binding to the TP53 promoter in alternative dimer configurations.

GROWTH INHIBITORY EFFECT OF HOUTTUYNIA CORDATA EXTRACT ON STREPTOCOCCUS MUTANS.

Houttuynia cordata is an herbal plant distributed throughout Asia. H. cordata has many bioactive properties, including antibacterial properties. The antibacterial effects of H. cordata on S. mutans remain unknown. Therefore, we treated S. mutans with 1, 3, 5, 10, 20, 30, or 40 mg/mL H. cordata extract at 37°C for 24 h. The antibacterial effect of H. cordata against S. mutans was confirmed using colony forming unit assay and disk diffusion assays. The results of the cell concentration assay demonstrated that H. cordata inhibited the growth of S. mutans in a dose-dependent manner. Prominent growth inhibition was observed after treatment with 10 mg/mL H. cordata extract, and these findings were statistically significant. In addition, no colonies of S. mutans were detected after treatment with 40 mg/mL H. cordata. Disk diffusion assays revealed that 20 mg/mL of H. cordata created a zone of growth inhibition of 11 mm. Therefore, our findings suggest the possibility of using H. cordata in the treatment and prevention of dental caries.

ANTIBACTERIAL EFFECT OF CRATAEGUS PINNATIFIDA EXTRACT AGAINST ENTROCOCCUS FAECALIS A ROOT CANAL DISEASE-CAUSING BACTERIA.

Studies have substantiated the anti-inflammatory and anti-thrombotic effects of (C. pinnatifida); however, research on its antibacterial activity using organic solvent remains limited. Therefore, in this study, we aimed to validate the antibacterial activity of C. pinnatifida as a natural extract against Enterococcus faecalis (E. faecalis), a multidrug-resistant bacterium. E. faecalis was treated with different concentrations of C. pinnatifida to determine the optimal concentration for the most effective antibacterial effect. Fifteen different concentrations were applied for 6 and 24 h. The experimental method centered on confirming antibacterial activity using colony-forming units. The experimental results demonstrated a proportional increase in antibacterial activity with elevated C. pinnatifida concentration. Notably, 99.99% and 100% antibacterial activity were observed at 10 mg/mL and 40 mg/mL concentrations, respectively. Our results suggest that C. pinnatifida holds potential as an antibacterial agent against the multidrug-resistant E. faecalis.

Fibrillin2 in chloroplast plastoglobules participates in photoprotection and jasmonate-induced senescence.

Fibrillins (FBNs) are the major structural proteins of plastoglobules (PGs) in chloroplasts. PGs are associated with defense against abiotic and biotic stresses, as well as lipid storage. Although FBN2 is abundant in PGs, its independent function under abiotic stress has not yet been identified. In this study, the targeting of FBN2 to PGs was clearly demonstrated using an FBN2-YFP fusion protein. FBN2 showed higher expression in green photosynthetic tissues and was upregulated at the transcriptional level under high-light stress. The photosynthetic capacity of fbn2 knockout mutants generated using CRISPR/Cas9 technology decreased rapidly compared with that of wild-type (WT) plants under high-light stress. In addition to the photoprotective function of FBN2, fbn2 mutants had lower levels of plastoquinone-9 and plastochromanol-8. The fbn2 mutants were highly sensitive to methyl jasmonate (MeJA) and exhibited root growth inhibition and a pale-green phenotype due to reduced chlorophyll content. Consistently, upon MeJA treatment, the fbn2 mutants showed faster leaf senescence and more rapid chlorophyll degradation with decreased photosynthetic ability compared with the WT plants. The results of this study suggest that FBN2 is involved in protection against high-light stress and acts as an inhibitor of jasmonate-induced senescence in Arabidopsis (Arabidopsis thaliana).

Transition readiness in middle and older adolescents with asthma and associated factors: a descriptive study.

OBJECTIVE: Asthma is the leading chronic health condition in adolescents, yet little is known about adolescents' readiness to transition into adult-focused care. This study examines transition readiness in middle and late adolescents with asthma. METHODS: This cross-sectional descriptive study was conducted in 2019 with 41 adolescents (16-20 years old) with asthma. Transition readiness was measured with the 20-item Transition Readiness Assessment Questionnaire (TRAQ), which comprises five subscales. RESULTS: Slight majorities of the sample comprised females (58.5%) and minority adolescents (56%). The sample's overall mean score on the TRAQ was 3.89 (±0.63; possible range 1-5). Managing daily activities was associated with talking with providers subscale (r = 0.36; p < 0.01), but not with other TRAQ subscales. Females reported greater scores for managing medications than did males, with mean = 4.2 vs. 3.6 (t = -2.15, p = 0.04). Transition readiness did not differ by race or health insurance type. However, scores on arranging transportation to medical appointments were lower among minority adolescents than whites (4.17 vs. 4.8, t = 2.56, p = 0.01). Better asthma control was associated with higher scores on talking with providers (r = -0.42; p < 0.01), but not with other subscale domains. CONCLUSIONS: Overall transition readiness is suboptimal in middle to late adolescents with asthma. Adolescents' capacity to manage daily activities is not a valid indicator of transition readiness in disease self-management. It is important to consider gender, race, and asthma control in understanding transition readiness in adolescents with asthma.

Sensorial saturation improves infants' procedure-related pain behaviour in the cardiac intensive care unit: A quasi-experimental study.

BACKGROUND: Painful procedures are unavoidable when providing critical care to infants in intensive care units. These adverse experiences during infancy can lead to later hyperalgesia and poor neurodevelopmental outcomes. Thus, appropriate interventions are required to relieve infant pain during these procedures. OBJECTIVES: This study evaluated the effectiveness of sensorial saturation in reducing pain for infants during jugular central venous catheter removal procedures in intensive care units. METHODS: This study involved a quasi-experimental, repeated-measures design. Data were collected from participants sequentially recruited from April to June 2019 (control period) and July to September 2019 (experimental period). Participants included 78 infants younger than 1 year with congenital heart disease. The control group (n = 38) received a general nursing intervention using swaddling, a common child-care practice that consists of wrapping infants to restrict movements, whereas the experimental group (n = 40) received sensorial saturation using oral sugar, body massage, and verbal interaction. Infants' physiological reactions to procedural pain were measured by changes in heart rate, oxygen saturation, and respiratory rate. Infants' procedural pain and behavioural indicators were measured using the Modified Behavioural Pain Scale. Data were analysed using descriptive statistics, independent t-tests, χ2 tests, and repeated-measures analysis of variance. RESULTS: Compared with the control group, the experimental group had lower heart rates (F = 53.15, p < .001), respiratory rates (F = 15.19, p < .001), and behavioural pain scores (F = 45.21, p < .001), both during and after the procedure. CONCLUSIONS: Sensorial saturation can be used as a nursing intervention in infants. Given the many invasive procedures that are part of infant clinical care, sensorial saturation may be a safe analgesic alternative. The findings of this study could lead to the development of evidence-based clinical practice guidelines for the nonpharmacological management of acute pain in infants.

Single-cell RNA-sequencing of Nicotiana attenuata corolla cells reveals the biosynthetic pathway of a floral scent.

High-throughput single-cell RNA sequencing (scRNA-Seq) identifies distinct cell populations based on cell-to-cell heterogeneity in gene expression. By examining the distribution of the density of gene expression profiles, we can observe the metabolic features of each cell population. Here, we employ the scRNA-Seq technique to reveal the entire biosynthetic pathway of a flower volatile. The corolla of the wild tobacco Nicotiana attenuata emits a bouquet of scents that are composed mainly of benzylacetone (BA). Protoplasts from the N. attenuata corolla limbs and throat cups were isolated at three different time points, and the transcript levels of > 16 000 genes were analyzed in 3756 single cells. We performed unsupervised clustering analysis to determine which cell clusters were involved in BA biosynthesis. The biosynthetic pathway of BA was uncovered by analyzing gene co-expression in scRNA-Seq datasets and by silencing candidate genes in the corolla. In conclusion, the high-resolution spatiotemporal atlas of gene expression provided by scRNA-Seq reveals the molecular features underlying cell-type-specific metabolism in a plant.

Structure-activity relationship analysis of novel GSPT1 degraders based on benzotriazinone scaffold and its antitumor effect on xenograft mouse model.

Molecular glue degraders, such as lenalidomide and pomalidomide, bind to cereblon (CRBN) E3 ligase and subsequently recruit neosubstrate proteins, Ikaros (IKZF1) and Aiolos (IKZF3), for the ubiquitination-proteasomal degradation process. In this study, we explored structure-activity relationship analysis for novel GSPT1 degraders utilizing a benzotriazinone scaffold previously discovered as a novel CRBN binder. In particular, we focused on the position of the ureido group on the benzotriazinone scaffold, substituent effect on the phenylureido group, and methyl substitution on the benzylic position of benzotriazinone. As a result, we identified 34f (TD-522), which exhibits strong anti-proliferative effects in both KG-1 (EC50 = 0.5 nM) and TMD-8 (EC50 = 5.2 nM) cell lines. Compound 34f effectively induced GSPT1 degradation with a DC50 of 0.269 nM and Dmax of >95 % at 10 nM concentration in KG-1 cells. An in vivo xenograft study showed that compound 34f effectively suppressed TMD8-driven tumor growth, suggesting a potential role in the development of novel GSPT1 degraders.

Crystal structures of human NSDHL and development of its novel inhibitor with the potential to suppress EGFR activity.

NAD(P)-dependent steroid dehydrogenase-like (NSDHL), an essential enzyme in human cholesterol synthesis and a regulator of epidermal growth factor receptor (EGFR) trafficking pathways, has attracted interest as a therapeutic target due to its crucial relevance to cholesterol-related diseases and carcinomas. However, the development of pharmacological agents for targeting NSDHL has been hindered by the absence of the atomic details of NSDHL. In this study, we reported two X-ray crystal structures of human NSDHL, which revealed a detailed description of the coenzyme-binding site and the unique conformational change upon the binding of a coenzyme. A structure-based virtual screening and biochemical evaluation were performed and identified a novel inhibitor for NSDHL harboring suppressive activity towards EGFR. In EGFR-driven human cancer cells, treatment with the potent NSDHL inhibitor enhanced the antitumor effect of an EGFR kinase inhibitor. Overall, these findings could serve as good platforms for the development of therapeutic agents against NSDHL-related diseases.

Dynamic changes in electrocardiographic findings between initial and follow-up electrocardiography: The role of the T/QRS ratio.

BACKGROUND: Dynamic changes in electrocardiographic findings between initial and follow-up electrocardiograms (ECGs) have rarely been studied for disease severity and differential diagnosis in non-ST elevation acute coronary syndrome. We aimed to determine whether the changes in staple variables on ECG can assist in distinguishing between neuropsychiatric or gastrointestinal disorders (mild non-ischemic disorders), heart failure, and NSTE-ACS (non-ST elevation acute coronary syndrome). METHODS: This retrospective study enrolled 1279 patients who presented with ischemic symptoms; were diagnosed with NSTE-ACS, acute heart failure, and mild disorders; and underwent echocardiography and coronary angiography. After performing propensity matching of the ECG follow-up interval, 184 patients with symptom onset within 48 h were included and analyzed. RESULTS: As a discriminator for NSTE-ACS, the maximum change in the T/QRS ratio in two contiguous leads was superior to the maximum change in ST segment depression, T wave inversion, and ST/T ratio. ECGs of patients with NSTE-ACS and heart failure showed a tendency to increase and decrease the T/QRS ratio change, respectively. Compared with regional wall motion abnormality, the change in troponin I/h and the maximum change in ST segment depression and T-wave inversion, the most deviated T/QRS ratio change from 1 (>1.5 or < 0.5) in two contiguous leads was the most significant discriminator for disease severity and differential diagnosis (standardized ß = 0.545, p < 0.001). CONCLUSION: The maximum changes in the T/QRS ratio in two contiguous leads can assist in distinguishing disease severity and acute mimicking disease such as acute heart failure in patients with suspected ACS.

NRF2/ARE pathway negatively regulates BACE1 expression and ameliorates cognitive deficits in mouse Alzheimer's models.

BACE1 is the rate-limiting enzyme for amyloid-ß peptides (Aß) generation, a key event in the pathogenesis of Alzheimer's disease (AD). By an unknown mechanism, levels of BACE1 and a BACE1 mRNA-stabilizing antisense RNA (BACE1-AS) are elevated in the brains of AD patients, implicating that dysregulation of BACE1 expression plays an important role in AD pathogenesis. We found that nuclear factor erythroid-derived 2-related factor 2 (NRF2/NFE2L2) represses the expression of BACE1 and BACE1-AS through binding to antioxidant response elements (AREs) in their promoters of mouse and human. NRF2-mediated inhibition of BACE1 and BACE1-AS expression is independent of redox regulation. NRF2 activation decreases production of BACE1 and BACE1-AS transcripts and Aß production and ameliorates cognitive deficits in animal models of AD. Depletion of NRF2 increases BACE1 and BACE1-AS expression and Aß production and worsens cognitive deficits. Our findings suggest that activation of NRF2 can prevent a key early pathogenic process in AD.

Health behaviors in adolescent survivors of cancer: An integrative review.

PROBLEM: Adolescent survivors of cancer face an increased risk of chronic health conditions that can be improved by healthy behaviors. The purpose of this review is to synthesize the extent/prevalence of health behaviors (physical activity [PA], diet, human papillomavirus [HPV] vaccination, alcohol use, smoking, marijuana use, and unprotected sex) compared to age-matched peers and factors associated with those behaviors. ELIGIBILITY CRITERIA: Four databases (PubMed, EMBASE, PsycINFO, and CINAHL) were searched for peer-reviewed primary studies published since 2000 including adolescents aged 11-20 years-old, who had completed cancer treatments. Studies reporting at least one behavior were included. SAMPLE: Of 1979 articles reviewed, 27 studies reporting any of the following health behaviors were included: PA, diet, HPV vaccination, alcohol use, smoking, marijuana use, or unprotected sex. RESULTS: Adolescent survivors of cancer engaged less in health risk behaviors (alcohol use, smoking, marijuana use, and unprotected sex) or health-promoting behaviors (PA, healthy diet, and HPV vaccination) compared to age-matched peers. Their health behaviors were associated with psychosocial factors including emotional discomfort (e.g., feeling depressed or nervous) and influence of others (family, friends, and healthcare providers). CONCLUSIONS: Although health risk behaviors in adolescent survivors of cancer are not as prevalent as in their peers, the survivors' low engagement in health-promoting behaviors including PA, diet, and HPV vaccination is concerning. IMPLICATIONS: More research is needed to develop and evaluate interventions to improve health-promoting behaviors in adolescent survivors of cancer by strengthening or facilitating psychosocial resources.

Evaluation of Cytotoxic and Antibacterial Effect of Methanolic Extract of Paeonia lactiflora.

BACKGROUND AND OBJECTIVES: Bacterial antibiotics have had several side effects. Therefore, interest in natural substances with less side effects is increasing these days. Paeonia lactiflora, the root of Paeonia lactiflora, is used as a raw material for medicines. In this study, we investigated the antibacterial effect and the cytotoxicity of Paeonia lactiflora extract. MATERIALS AND METHODS: For cytotoxicity, MTT analysis according to ISO 10993-5 was performed. The antibacterial test of the Paeonia lactiflora was determined from bacterial viability, Inhibition zone test, CFU (colony forming unit) and SEM (scanning electron microscope). To confirm the antibacterial component of Paeonia lactiflora, the content of flavonoids and polyphenols was analyzed. RESULTS: Our results showed that Paeonia lactiflora extract contained flavonoids and polyphenols, which exhibited antimicrobial activity against Streptococcus mutans (S. mutans) and Candida albicans (C. ablicans). Further, the cytotoxicity of Paeonia lactiflora extract was low. CONCLUSIONS: We believe that our study makes a significant contribution to the literature because it demonstrates that Paeonia lactiflora extract can be used as an antibiotic.

Surface Characterization, Antimicrobial Activity of Nonthermal Atmospheric-Pressure Plasma Jet on Polyvinyl Siloxane Impression Materials.

Background and Objectives The antimicrobial efficacy of a nonthermal atmospheric-pressure plasma jet (NAPPJ) on dental impression materials was investigated. Materials and Methods Type 3 polyvinyl siloxane was used as the impression material, and air and nitrogen NAPPJ were applied. The antibacterial effect of the NAPPJ was measured using the number of colony-forming units (CFUs) and scanning electron microscopy (SEM) images of Streptococcus mutans. Surface chemical characteristics of the impression material were examined using X-ray photoelectron spectroscopy (XPS) and contact angle measurement. Additionally, physical properties were analyzed through surface roughness measurement, detail reproduction, and strain-in-compression test. Results Compared with the control group, the plasma treatment group showed ruptured bacteria membranes, destroyed bacteria structures, a significant reduction in the number of CFUs, and a significantly reduced contact angle. Further, XPS analysis showed that their surface was significantly richer in hydroxyl groups. The surface roughness, detail reproduction, and strain-in-compression results indicated no significant differences between the plasma treatment and control groups. NAPPJ treatment could remove bacteria from polyvinyl siloxane dental impression materials without changing the surface's physical properties. Conclusion Therefore, it is considered a promising method for disinfection.

Thermodynamic Models for Assembly of Intrinsically Disordered Protein Hubs with Multiple Interaction Partners.

Prevalent in diverse protein interactomes, intrinsically disordered proteins or regions (IDPs or IDRs) often drive assembly of higher-order macromolecular complexes, using multiple target-binding motifs. Such IDP hubs are suggested to process various cellular signals and coordinate relevant biological processes. However, the mechanism of assembly and functional regulation of IDP hubs remains elusive due to the challenges in dissecting their intricate protein-protein interaction networks. Here we present basic thermodynamic models for the assembly of simple IDP hubs with multiple target proteins, constructing partition functions from fundamental binding parameters. We combined these basic functions to develop advanced thermodynamic models to analyze the assembly of the Nup153 hubs interacting with multiple karyopherin ß1 (Kap) molecules, critical components of nucleocytoplasmic transport. The thermodynamic analysis revealed a complex organization of the Kap binding sites within the C-terminal IDR of Nup153 including a high-affinity 1:1 interaction site and a series of low-affinity sites for binding of multiple Kaps with negative cooperativity. The negative cooperativity arises from the overlapping nature of the low-affinity sites where Kap occupies multiple dipeptide motifs. The quantitative dissection culminated in construction of the Nup153 hub ensemble, elucidating how distribution among various Kap-bound states is modulated by Kap concentration and competing nuclear proteins. In particular, the Kap occupancy of the IDR can be fine-tuned by varying the location of competition within the overlapping sites, suggesting coupling of specific nuclear processes to distinct transport activities. In general, our results demonstrate the feasibility and a potential mechanism for manifold regulation of IDP functions by diverse cellular signals.

Change in T/QRS ratio can be a supplementary diagnostic tool in predicting coronary artery disease in patients with NSTEMI.

BACKGROUND: Changes in the electrocardiographic findings, namely the ratio T sum to QRS sum (T/QRS ratio), between the initial electrocardiogram (ECG) and the baseline ECG have rarely been investigated in patients with non-ST elevation myocardial infarction (NSTEMI). Thus, we aimed to determine whether changes in various parameters on ECG, including T/QRS ratio, can assist in distinguishing between coronary artery disease (CAD) and NSTEMI without CAD with low to moderate risk. METHODS: This retrospective study enrolled 2572 patients who presented ischemic symptoms, who were diagnosed with NSTEMI, and who underwent coronary angiography. Overall, 388 patients had prior ECG and echocardiography data available; 110 patients were included after excluding patients with other cardiac diseases except CAD. The population divided into two groups: a coronary stenosis group (n = 78); normal coronary group (n = 32) were analyzed. RESULTS: We found that acute dynamic change in the most deviated T/QRS ratio in each region of leads of initial ECG from those of remote/recent ECG was an extremely strong predictor of acute CAD (odds ratio, 110; p < .001) compared to that of initial serum troponin I levels, new-onset regional wall motion abnormalities, and new-onset T inversion or ST depression. T/QRS ratio change > 1.5 or <0.5 times in injured regional leads was a significant predictor of CAD. CONCLUSION: Change in the most deviated T/QRS ratio in the regional leads on initial ECG from the T/QRS ratio in the same lead on remote ECG can assist in predicting CAD risk between patients with CAD and patients with no CAD in NSTEMI.

Comparative Transcriptome-Based Mining of Senescence-Related MADS, NAC, and WRKY Transcription Factors in the Rapid-Senescence Line DLS-91 of Brassica rapa.

Leaf senescence is a developmental process induced by various molecular and environmental stimuli that may affect crop yield. The dark-induced leaf senescence-91 (DLS-91) plants displayed rapid leaf senescence, dramatically decreased chlorophyll contents, low photochemical efficiencies, and upregulation of the senescence-associated marker gene BrSAG12-1. To understand DLS molecular mechanism, we examined transcriptomic changes in DLS-91 and control line DLS-42 following 0, 1, and 4 days of dark treatment (DDT) stages. We identified 501, 446, and 456 DEGs, of which 16.7%, 17.2%, and 14.4% encoded TFs, in samples from the three stages. qRT-PCR validation of 16 genes, namely, 7 MADS, 6 NAC, and 3 WRKY, suggested that BrAGL8-1, BrAGL15-1, and BrWRKY70-1 contribute to the rapid leaf senescence of DLS-91 before (0 DDT) and after (1 and 4 DDT) dark treatment, whereas BrNAC046-2, BrNAC029-2/BrNAP, and BrNAC092-1/ORE1 TFs may regulate this process at a later stage (4 DDT). In-silico analysis of cis-acting regulatory elements of BrAGL8-1, BrAGL42-1, BrNAC029-2, BrNAC092-1, and BrWRKY70-3 of B. rapa provides insight into the regulation of these genes. Our study has uncovered several AGL-MADS, WRKY, and NAC TFs potentially worthy of further study to understand the underlying mechanism of rapid DLS in DLS-91.