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Heat stress significantly impacts plant growth and productivity by causing protein misfolding, oxidative damage, and disruptions in cellular homeostasis. This stress is particularly critical during flowering and grain-filling stages, where even brief exposure to high temperatures can result in severe yield losses. Plants have evolved complex molecular mechanisms to mitigate heat stress, including the activation of heat shock proteins and the ubiquitin-proteasome system to maintain protein quality and cellular function. Understanding these processes is essential for breeding stress-resistant crops capable of adapting to climate change. In this study, we characterized TaCEU, a wheat carboxy-terminal Hsp70-interacting protein and E3 ubiquitin ligase, with a conserved structure across plant species. TaCEU is highly upregulated under heat and ABA stress and plays a pivotal role in maintaining protein homeostasis during heat stress. We confirmed TaCEU's role as an E3 ligase through in vitro ubiquitination assays, demonstrating its ability to form polyubiquitin chains. Subcellular localization studies confirmed TaCEU's presence in the nucleus and plasma membrane, suggesting functional versatility. Interaction studies identified TaAP and TaCHSP70 as key partners, with TaCEU and TaCHSP70 collaboratively targeting TaAP for degradation via the UPS proteasome system. Functional analysis of TaCEU-overexpressing Arabidopsis revealed enhanced thermotolerance, including increased root length, fresh weight, and antioxidant enzyme activity under heat stress. Transcript analysis further indicated the upregulation of heat stress-responsive genes, highlighting TaCEU's critical role in stress adaptation. These findings establish TaCEU as a key regulator of thermotolerance and protein homeostasis. TaCEU regulates protein stability and enhancing thermotolerance, making it a promising target for developing heat-resistant crops to address the challenges of climate change., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Background: The adductor canal block is a well-known procedure for controlling postoperative pain after medial malleolus fracture surgery. Continuous nerve block is a viable option for blocking pain for a longer period although the literature on this subject is scarce. Therefore, this study aimed to compare continuous adductor canal block (cACB) group to single-injection adductor canal block (sACB) group in those with bimalleolar or trimalleolar ankle fractures. The procedure was performed in addition to a continuous sciatic nerve block for postoperative pain relief and patient satisfaction., Methods: The study included 57 patients who had bimalleolar or trimalleolar ankle fractures and underwent open reduction and internal fixation between August 2016 and June 2018. Each patient received a continuous sciatic nerve block and was divided into two groups: those who received cACB and those who received sACB. Each postoperative pain was scored at 4, 8, 12, 24, 48, and 72 h after surgery. Additionally, the consumption of rescue medications and patient satisfaction were evaluated., Results: The two groups displayed no disparity in medial side ankle pain at 4 h and 8 h after surgery, but significantly higher pain in the sACB group at 12, 24, 48, and 72 h after surgery. However, there was no difference in the pain at the lateral side of ankle and consumption of rescue medication. In addition, the cACB group showed more satisfaction than the sACB group did., Conclusion: CACB is better than sACB in terms of postoperative pain control and patient satisfaction. cACB can be used for postoperative pain control in ankle fractures involving the medial malleolus., Level of Evidence: Prospective Randomized Controlled Trial, Level 2., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as potential conflicts of interest., (Copyright © 2024 The Japanese Orthopaedic Association. Published by Elsevier B.V. All rights reserved.)

The initiation of transition to flowering is carefully managed by endogenous and environmental cues, which is critical for flowering plant reproductive success. Here, we found that wheat RING-type E3 ligase TaFRFP was highly expressed from the double ridge to degeneration stage (WS2.5-WS9). TaFRFP is localized in the nucleus and has E3 ligase activity in vitro. TaFRFP overexpression in Arabidopsis resulted in an early flowering phenotype, but to a lesser extent, under short-day conditions. Under the SA-treated condition, overexpression of TaFRFP shows higher root growth and has more accumulation of SA contents. A proteomic comparison revealed that the amount of FRL4A protein, a FRIGIDA LIKE 4 A, was considerably lower in SA-treated TaFRFP seedlings compared to normal condition. We further found that TaFRFP directly interacts with FRL4A in the nucleus and recruits it to the FLC locus in Arabidopsis. Moreover, an ubiquitination assay showed that TaFRPF physically interact and ubiquitinates TaFRL as a substrate. Our findings support the concept that the TaFRFP E3 ligase works as a positive regulator, and that the ubiquitination of its substrate proteins plays a significant role in controlling flowering time via an SA-dependent pathway., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Deep vein thrombosis (DVT) is a common complication in orthopedic surgery and has the potential to lead to fatal complications such as pulmonary thromboembolism. However, the precise incidence and risk factors for DVT in the foot and ankle fields remain unclear. This study aimed to analyze the prevalence of DVT and identify its risk factors after foot and ankle surgery., Methods: Between September 2020 and July 2023, screening duplex ultrasonography was performed on 278 patients who underwent foot and ankle surgery and required immobilization. The findings from screening duplex ultrasonography were assessed in conjunction with the symptoms present at the time of diagnosis. Heterogeneous demographic data that could serve as potential risk factors for DVT, including diagnosis, body mass index, and other medical histories, were examined alongside pertinent surgery-related data, such as tourniquet time., Results: Among the 278 individuals, DVT occurred in 41 patients (14.7%). Among these, 92.7% originated at the calf level and the majority were asymptomatic. The cases originating above the calf accounted for 3 cases, representing 7.3% of patients diagnosed with DVT (1.1% of the entire screened population). Acute trauma, history of previous DVT, and old age were identified as statistically significant risk factors for DVT occurrence, with odds ratios of 2.44 ( p = 0.04), 6.40 ( p = 0.02), and 1.16 ( p = 0.03), respectively., Conclusions: After foot and ankle surgery, DVT occurred in 14.7% of cases. Acute trauma, history of DVT, and old age were identified as risk factors for DVT. These findings highlight the necessity of careful monitoring and appropriate prophylactic interventions for high-risk patients. Further investigation is required to determine effective prophylactic strategies for this patient population., Competing Interests: CONFLICT OF INTEREST: No potential conflict of interest relevant to this article was reported., (Copyright © 2024 by The Korean Orthopaedic Association.)

Biomaterials can act as pro- or anti-inflammatory agents. However, effects of biomaterials crystallinity on immune responses are poorly understood. We demonstrate that the adjuvant-like behaviour of covalent organic framework (COF) biomaterial is dependent on its crystallinity. COF crystallinity is inversely correlated with the activation of mouse and human dendritic cells (DC), but with antigen presentation by mouse DCs only. Amorphous COFs upregulates NFkB, TNF, and RIG-I signalling pathways, as well as the chemotaxis-associated gene Unc5c, when compared to crystalline COFs. Meanwhile, Unc5c inhibition disrupts the correlation between crystallinity and DC activation. Furthermore, COFs with the lowest crystallinity admixed with chicken ovalbumin (OVA) antigen prevent OVA-expressing B16F10 tumour growth in 60% of mice, with this protection associated with the induction of antigen-specific, pro-inflammatory T cell. The lowest crystalline COFs admixed with TRP2 antigen can also prevent non-immunogenic YUMM1.1 tumour growth in 50% of mice. These findings demonstrate that the crystallinity of biomaterials is an important aspect to consider when designing immunotherapy for pro- or anti-inflammatory applications., Competing Interests: Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Background: Hindfoot malalignment can cause various foot and ankle problems. For better surgical performance and correction of hindfoot malalignments, reliable intraoperative determination of hindfoot alignment is essential. However, there is no standard method for the intraoperative assessment of hindfoot alignment. We devised an intraoperative modified Méary posteroanterior (IOPPA) view to assess intraoperative hindfoot alignment. This study aimed to compare this intraoperative method with other radiographic hindfoot alignment measurements., Methods: Thirty-seven patients (47 feet) with various foot and ankle conditions scheduled to undergo surgery were prospectively recruited. Before surgery, the Saltzman, long axial, and modified Méary views were taken in a controlled and standardized fashion. IOPPA views were obtained under simulated weight bearing conditions using C-arm fluoroscopy in the operating room before surgery. The relationship between the IOPPA view and the three radiographic hindfoot alignments was evaluated using Pearson's correlation., Results: The mean hindfoot alignment angle was varus 3.50° (CI, varus 1.91 to 5.08) on the Saltzman view, varus 2.00° (CI, varus 0.60 to 3.39) on the long axial view, varus 0.13° (CI, valgus 1.41 to varus 1.67) on the modified Méary view, and varus 1.32° (CI, valgus 0.02 to varus 2.65) on IOPPA view. The IOPPA view and the three other hindfoot alignment views were found to be significantly correlated (r = 0.60 for the Saltzman view, r = 0.50 for the long axial view, r = 0.71 for the modified Méary view, P < .05). The intraobserver ICC (Intraclass Correlation Coefficient) value was 0.974 and interobserver ICC (Intraclass Correlation Coefficient) value was 0.988 for the IOPPA view (P < .001)., Conclusion: There was a statistically significant correlation between the IOPPA view and the other three hindfoot alignment views. We also found that interobserver and intraobserver ICC values were excellent. This study proposes that the IOPPA view can be used as a reliable intraoperative assessment tool for hindfoot alignment., Level of Evidence: Prospective study., Competing Interests: Declaration of competing interest The authors received no financial support for the research, authorship, and/or publication of this article., (Copyright © 2023 The Japanese Orthopaedic Association. Published by Elsevier B.V. All rights reserved.)

Key Message: 111 PHD genes were newly identified in rye genome and ScPHD5's role in regulating cold tolerance and flowering time was suggested. Plant homeodomain (PHD)-finger proteins regulate the physical properties of chromatin and control plant development and stress tolerance. Although rye (Secale cereale L.) is a major winter crop, PHD-finger proteins in rye have not been studied. Here, we identified 111 PHD genes in the rye genome that exhibited diverse gene and protein sequence structures. Phylogenetic tree analysis revealed that PHDs were genetically close in monocots and diverged from those in dicots. Duplication and synteny analyses demonstrated that ScPHDs have undergone several duplications during evolution and that high synteny is conserved among the Triticeae species. Tissue-specific and abiotic stress-responsive gene expression analyses indicated that ScPHDs were highly expressed in spikelets and developing seeds and were responsive to cold and drought stress. One of these genes, ScPHD5, was selected for further functional characterization. ScPHD5 was highly expressed in the spike tissues and was localized in the nuclei of rye protoplasts and tobacco leaves. ScPHD5-overexpressing Brachypodium was more tolerant to freezing stress than wild-type (WT), with increased CBF and COR gene expression. Additionally, these transgenic plants displayed an extremely early flowering phenotype that flowered more than two weeks earlier than the WT, and vernalization genes, rather than photoperiod genes, were increased in the WT. RNA-seq analysis revealed that diverse stress response genes, including HSPs, HSFs, LEAs, and MADS-box genes, were also upregulated in transgenic plants. Our study will help elucidate the roles of PHD genes in plant development and abiotic stress tolerance in rye., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Freezing temperature during overwintering often kills plants; plants have thus, developed a defense mechanism called 'cold acclimation', in which a number of genes are involved in increasing cell protection and gene expression. Mitogen-activated protein kinase (MAPK) controls proteins' activities by phosphorylation and is involved in numerous metabolic pathways. In this study, we identified the protein interaction between TaMAPK3 and the proteins in the cold response pathway, ICE41, ICE87, and CBFIVd-D9. The subcellular localization and bimolecular fluorescence complement (BiFC) assays revealed that these proteins interact in the nucleus or in the plasma membrane. Furthermore, MAPK3-mediated phosphorylation of ICE41, ICE87, and CBFIVd-D9 was verified using an in vitro phosphorylation assay. TaMAPK3-overexpressing transgenic Brachypodium showed a lower survival rate upon freezing stress and lower proline content during cold acclimation, compared to wild-type plants. Furthermore, cold response gene expression analysis revealed that the expression of these genes was suppressed in the transgenic lines under cold treatment. It was further elucidated that MAPK3 mediates the degradation of ICE and CBF proteins, which implies the negative impact of MAPK3 on the freezing tolerance of plants. This study will help to elucidate the molecular mechanisms of cold tolerance and the activity of MAPK3 in wheat., Competing Interests: Declaration of competing Interest The authors declare that they have no competing interests., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Background: Diabetic foot (DMF) complications are common and are increasing in incidence. Risk factors related to wound complications are yet to be established after trans-tibial amputation under the diagnosis of DMF infection., Aim: To analyze the prognosis and risk factors related to wound complications after transtibial amputation in patients with diabetes., Methods: This retrospective cohort study included seventy-two patients with DMF complications who underwent transtibial amputation between April 2014 and March 2023. The groups were categorized based on the occurrence of wound complications, and we compared demographic data between the complication group and the non-complication group to analyze risk factors. Moreover, a multivariate logistic regression analysis was performed to identify risk factors., Results: The average follow-up period was 36.2 months. Among the 72 cases, 31 (43.1%) had wound complications. Of these, 12 cases (16.7%) received further treatment, such as debridement, soft tissue stump revision, and re-amputation at the proximal level. In a group that required further management due to wound complications after transtibial amputation, the hemoglobin A1c (HbA1c) level was 9.32, while the other group that did not require any treatment had a 7.54 HbA1c level. The prevalence of a history of kidney transplantation with wound complications after transtibial amputation surgery in DMF patients was significantly greater than in cases without wound complications ( P = 0.02). Other factors did not show significant differences., Conclusion: Approximately 43.1% of the patients with transtibial amputation surgery experienced wound complications, and 16.7% required additional surgical treatment. High HbA1c levels and kidney transplant history are risk factors for postoperative wound complications., Competing Interests: Conflict-of-interest statement: The authors received no financial support for the research, authorship, and/or publication of this article., (©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.)

Introduction: This case report describes the diagnosis of a glomus tumor in the second toe of a 38-year-old female, followed by surgical treatment utilizing a transungual approach to preserve the nail. This study highlights the diagnostic challenges and surgical strategies to treat such tumors while preserving nail integrity., Patient Concerns: Pain occurred once a week, but over time, it increased, and just before seeking medical attention, she experienced pain more than 5 times a day. The pain worsened when cold water touched her toe., Diagnosis: We observed a slight hump indicating nail plate deformity, and the patient exhibited severe pinpoint tenderness (positive Love test) in the affected area. Color duplex ultrasound was performed for further investigation, revealing a hypervascular hypoechoic nodule measuring 0.5 cm in size at the nail bed of the right second toe., Intervention: The surgery was performed under digital nerve block anesthesia using a modified transungual nail-preserving approach for the excision of the glomus tumor., Outcomes: The pain that was reported prior to the surgery has improved postoperatively, and the recovery has been uneventful without any other complication., Conclusion: This paper provides a comprehensive examination of a rare glomus tumor in the second toe, elucidating both diagnostic intricacies and treatment modalities. It emphasizes the dual necessity of achieving total tumor excision while also considering aesthetic outcomes. The insights presented herein are intended to serve as valuable guidance for clinicians confronted with similar clinical scenarios, underlining the delicate interplay between effective tumor management and the preservation of cosmetic integrity., Competing Interests: The authors have no funding and conflicts of interest to disclose., (Copyright © 2024 the Author(s). Published by Wolters Kluwer Health, Inc.)

The transition of food crops from the vegetative to reproductive stages is an important process that affects the final yield. Despite extensive characterization of E3 ligases in model plants, their roles in wheat development remain unknown. In this study, we elucidated the molecular function of wheat TaATL1 (Arabidopsis thaliana Toxicos EN Levadura), which acts as a negative regulator of flowering time and cell division. TaATL1 amino acid residues contain a RING domain and exist mainly in a beta-turn form. The expression level of TaATL1 was highly reduced during the transition from vegetative to reproductive stages. TaATL1 is localized in the nucleus and exhibits E3 ligase activity. Transgenic Arabidopsis plants, in which the TaATL1 gene is constitutively overexpressed under the control of the cauliflower mosaic virus 35 S promoter, exhibited regulation of cell numbers, thereby influencing both leaf and root growth. Moreover, TaATL1 overexpression plants showed a late-flowering phenotype compared to wild-type (WT) plants. Following transcriptome analysis, it was discovered that 1661 and 901 differentially expressed genes were down- or up- regulated, respectively, in seedling stages between WT and TaATL1 overexpression. TaATL1 transcripts are involved in cell division, flowering, and signaling. Overall, our findings demonstrated that the regulatory mechanism of wheat TaATL1 gene plays a significant role in cell division-mediated flowering in Arabidopsis., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This animal study was aimed to evaluate the efficacy of new bone formation and volume maintenance according to the particle type and the collagen membrane function for grafted octacalcium phosphate (OCP) in rabbit calvarial defects. The synthetic bone substitutes were prepared in powder form with 90% OCP and granular form with 76% OCP, respectively. The calvarial defects were divided into four groups according to the particle type and the membrane application. All specimens were acquired 2 weeks (n = 5) and 8 weeks (n = 5) after surgery. According to the micro-CT results, the new bone volume increased at 2 weeks in the 76% OCP groups compared to the 90% OCP groups, and the bone volume ratio was significantly lower in the 90% OCP group after 2 weeks. The histomorphometric analysis results indicated that the new bone area and its ratio in all experimental groups were increased at 8 weeks except for the group with 90% OCP without a membrane. Furthermore, the residual bone graft area and its ratio in the 90% OCP groups were decreased at 8 weeks. In conclusion, all types of OCP could be applied as biocompatible bone graft materials regardless of its density and membrane application. Neither the OCP concentration nor the membrane application had a significant effect on new bone formation in the defect area, but the higher the OCP concentration, the less graft volume maintenance was needed.

High temperatures can significantly impact wheat growth and grain yields during the grain-filling stage. In this study, we identified genes that respond to high-temperature stress during the grain-filling stage. We also identified and characterized 24 novel genes of the DOG1 gene family in hexaploid wheat. Motif analysis and conserved domain search revealed substantial similarities among TaDOG1 family members. Phylogenetic analysis demonstrated the evolutionary conservation of the TaDOG1 family across various plant species. Tissue-specific expression profiling indicated consistent patterns, with TaDOG1 genes predominantly expressed in stem tissues. Only TaDOG1-1 exhibited enhanced expression, particularly during hard dough and ripening stages. TaDOG1-1 and TaDOG1-7 exhibited increased expression under heat stress during the grain-filling stage, indicating their heat-responsive nature. Cis-element analysis revealed potential regulatory motifs, suggesting the involvement of TaDOG1-1 and TaDOG1-7 in stress tolerance mechanisms. Yeast two-hybrid screening revealed interacting proteins, including stress-responsive and grain development-associated proteins. To understand the biological function, we overexpressed TaDOG1-1 in Arabidopsis plants and observed enhanced thermotolerance under basal heat stress. Under heat stress, the transgenic plants exhibited increased biomass and elevated expression levels of heat-responsive genes. Furthermore, TaDOG1-1-overexpressing plants showed improved survival rates under soil heat stress, along with a greater accumulation of antioxidant enzymes in leaves. In this study, the identification and functions of the DOG1 gene family provide valuable insights for developing genetic engineering strategies aimed at improving wheat yield under high-temperature stress., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Purpose: The aim of this study was to investigate the efficacy of photo-crosslinked gelatin methacryloyl (GelMa) hydrogel containing calcium phosphate nanoparticles (CNp) when applying different fabrication methods for bone regeneration., Methods: Four circular defects were created in the calvaria of 10 rabbits. Each defect was randomly allocated to the following study groups: 1) the sham control group, 2) the GelMa group (defect filled with crosslinked GelMa hydrogel), 3) the CNp-GelMa group (GelMa hydrogel crosslinked with nanoparticles), and 4) the CNp+GelMa group (crosslinked GelMa loaded with nanoparticles). At 2, 4, and 8 weeks, samples were harvested, and histological and micro-computed tomography analyses were performed., Results: Histomorphometric analysis showed that the CNp-GelMa and CNp+GelMa groups at 2 weeks had significantly greater total augmented areas than the control group ( P <0.05). The greatest new bone area was observed in the CNp-GelMa group, but without statistical significance ( P >0.05). Crosslinked GelMa hydrogel with nanoparticles exhibited good biocompatibility with a minimal inflammatory reaction., Conclusions: There was no difference in the efficacy of bone regeneration according to the synthesized method of photo-crosslinked GelMa hydrogel with nanoparticles. However, these materials could remain within a bone defect up to 2 weeks and showed good biocompatibility with little inflammatory response. Further improvement in mechanical properties and resistance to enzymatic degradation would be needed for the clinical application., Competing Interests: No potential conflict of interest relevant to this article was reported., (Copyright © 2023. Korean Academy of Periodontology.)

Histone methylation is actively involved in plant flowering time and is regulated by a myriad of genetic pathways that integrate endogenous and exogenous signals. We identified an F-box gene from wheat (Triticum aestivum L.) and named it TaF-box3. Transcript expression analysis showed that TaF-box3 expression was gradually induced during the floret development and anthesis stages (WS2.5-10). Furthermore, ubiquitination assays have shown that TaF-box3 is a key component of the SCF ubiquitin ligase complex. TaF-box3 overexpression in Arabidopsis resulted in an early flowering phenotype and different cell sizes in leaves compared to the WT. Furthermore, the transcript level of a flowering time-related gene was significantly reduced in TaF-box3 overexpressing plants, which was linked with lower histone H3 Lys4 trimethylation (H3K4me3) and H3 Lys36 trimethylation (H3K36me3). Overexpression of TaF-box3 in Arabidopsis was shown to be involved in the regulation of flowering time by demethylating FLC chromatin, according to ChIP experiments. Protein analysis confirmed that TaMETS interacts with TaF-box3 and is ubiquitinated and degraded in a TaF-box3-dependnent manner. Based on these findings, we propose that TaF-box3 has a positive role in flowering time, which leads to a better understanding of TaF-box3 physiological mechanism in Arabidopsis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this study., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The importance of oats has increased because of their high nutritional value and health benefits in the human diet. High-temperature stress during the reproductive growth period has a detrimental effect on grain morphology by changing the structure and concentration of several seed-storage proteins. DA1, a conserved ubiquitin-proteasome pathway component, plays an important role in regulating grain size by controlling cell proliferation in maternal integuments during the grain-filling stage. However, there have been no reports or studies on oat DA1 genes. In this study, we identified three DA1-like genes (AsDA1-2D, AsDA1-5A, and AsDA1-1D) using genome-wide analysis. Among these, AsDA1-2D was found to be responsible for high-temperature stress tolerance via a yeast thermotolerance assay. The physical interaction of AsDA1-2D with oat-storage-globulin (AsGL-4D) and a protease inhibitor (AsPI-4D) was observed using yeast two-hybrid screening. A subcellular localization assay revealed that AsDA1-2D and its interacting proteins are localized in the cytosol and plasma membrane. An in vitro pull-down assay showed that AsDA1-2D forms a complex with both AsPI-4D and AsGL-4D. An in vitro cell-free degradation assay showed that AsGL-4D was degraded by AsDA1-2D under high-temperature conditions and that AsPI-4D inhibited the function of AsDA1-2D. These results suggest that AsDA1-2D acts as a cysteine protease and negatively regulates oat-grain-storage-globulin under heat stress., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier GmbH. All rights reserved.)

Common wheat (Triticum aestivum) is a major cereal crop grown and consumed globally. Recent advances in sequencing technology have facilitated the exploration of large and repetitive genomes. Plant sucrose transporter (SUT) genes are vital components of energy transport systems that play prominent roles in various plant functions, such as signaling and stress regulation. In this study, we identified and analyzed five novel sucrose transporter genes in wheat. The wheat sucrose transporter genes were divided into five clades based on their phylogenetic relationships. Synteny analysis revealed that synteny in the genome is highly conserved between wheat and rye, barley, and Brachypodium. Furthermore, the cis-element analysis indicated that sucrose transporter genes might be regulated by light and some phytohormone-related transcriptional factors. Overall, plant tissue-specific gene expression revealed enhanced expression of the transporter genes in the root and stem, whereas they were differentially expressed under abiotic stress treatments (cold, heat, NaCl, PEG-6000, and sucrose). These results indicate that each TaSUT gene may play a crucial role in stabilizing plants under stress by actively regulating the energy demands of cells. The findings of this study may provide a basis for further research on sucrose transporters and their significant roles in plant energy metabolism as well as in abiotic stress response, signaling, and regulation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background: We aimed to find out whether the combined treatment of acupuncture and oral medication is more effective than sole oral medication in reducing pain and improving knee function at the end of treatment and after short-term period (4-6 weeks after treatment). Second, if it is effective, we investigated whether the effect surpasses the minimal clinically important difference., Methods: Articles published between January 1, 1992, and August 31, 2022, were searched in PubMed, Cochrane, and Embase. The PICO (population, intervention, comparison, and outcome) of this study are as follows: Population: knee osteoarthritis patients; Intervention: acupuncture (non-sham acupuncture) + oral medication (analgesic or non-steroidal anti-inflammatory drugs); Comparison: oral medication (analgesic or non-steroidal anti-inflammatory drugs); Outcome: visual analog scale (VAS) or Western Ontario and McMaster University (WOMAC) osteoarthritis index., Results: The combined treatment of oral medication and adjuvant acupuncture showed statistically significant improvement in VAS and WOMAC scores at the end of acupuncture treatment and short-term follow-up time (between 4 and 6 weeks after acupuncture). In addition, the degree of improvement of VAS and WOMAC index showed effects beyond minimal clinically important differences compared to pretreatment at both the end of acupuncture treatment and the short-term follow-up of acupuncture treatment., Conclusion: The existing evidence suggests that adjuvant acupuncture may play a role in the treatment of knee osteoarthritis. However, physicians should be aware of adverse effects such as hematoma in adjuvant acupuncture treatment., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2023 the Author(s). Published by Wolters Kluwer Health, Inc.)

Background: Fatigue and rotation time are key factors affecting the quality of cardiopulmonary resuscitation (CPR). This study aimed to investigate the effects of rotation time on CPR duration, and sex on chest compression quality., Methods: In this randomized crossover simulation study, we enrolled 100 paramedic students stratified by sex and randomly grouped into 28 male and 22 female pairs. In the 2- and 1-minute scenarios, 2 participants performed CPR for 20 minutes with a rotation cycle of 2 minutes and 1 minute, respectively. After taking a break, they changed over and performed CPR again for 20 minutes. The switching of roles was performed with the students positioned on opposite sides of the mannequin. For chest compression quality, a set was defined as CPR performed by 1 pair in a 2-minute scenario for 4 minutes. The quality of CPR in each set was compared between the 2 groups., Results: The 1-minute group showed significantly higher chest compression depth than the 2-minute group (54.0 [51.5-57.0] vs 52.5 [48.5-56.5] mm, P = .001, respectively). The female 2-minute group showed decreased chest compression depth over time, and the 1-minute group showed significantly increased chest compression depth during all sets except the 2nd set (54.0 [51.9-55.1] vs 50.5 [48.5-53.8] mm [P = .030], 52.3 [49.4-54.5] vs 50.8 [47.0-53.1] mm [P = .080], 52.8 [49.8-54.5] vs 48.8 [45.4-51.6] mm [P = .002], 51.5 [48.5-53.3] vs 48.3 [44.5-50.6] mm [P = .004], and 50.8 [48.9-54.1] vs 47.5 [44.6-50.1] mm [P = .001], respectively). The fatigue scores in the 2-minute group were significantly higher during sets 4 and 5 than those in the 1-minute group., Conclusion: When rescuer fatigue increases during prolonged CPR owing to their physical strength and skill levels, the rotation of rescuers every minute would be helpful in maintaining high-quality CPR., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2023 the Author(s). Published by Wolters Kluwer Health, Inc.)

Republic of Korea's suicide rate is the highest among Organization for Economic Co-operation and Development countries. In Republic of Korea, suicide is the leading cause of death among young people aged 10-19 years. This study aimed to identify changes in patients aged 10-19 years who visited the emergency department in Republic of Korea after inflicting self-harm over the past five years and to compare the situations before and after the outbreak of the COVID-19 pandemic. Analysis of government data revealed that the average daily visits per 100,000 were 6.25, 8.18, 13.26, 15.31, and 15.71 from 2016 to 2020, respectively. The study formed four groups for further analysis, with the population divided by sex and age (10-14 and 15-19 years old). The late-teenage female group showed the sharpest increase and was the only group that continued to increase. A comparison of the figures 10 months before and after the outbreak of the pandemic revealed a statistically significant increase in self-harm attempts by only the late-teenage female group. Meanwhile, visits (per day) in the male group did not increase, but the rates of death and ICU admission increased. Additional studies and preparations that account for age and sex are warranted.

Drought stress frequently results in significant reductions in crop production and yield. Plant U-box proteins (PUB) play a key role in the response to abiotic stress. Despite extensive characterization of PUB in model plants, their roles in wheat abiotic stress response remains unknown. In this study, we identified the physiological function of TaPUB4, a gene encoding the U-box and nuclear localization domains. The transcription level of TaPUB4 was induced by drought (mannitol) and abscisic acid. TaPUB4 displays E3 ubiquitin ligase activity and is located in the nucleus. Overexpression of TaPUB4 in Arabidopsis plants enhanced sensitivity with under ABA condition during early seedling developmental stages. In addition, the stomatal conductance of TaPUB4 was closer to that of WT under ABA conditions. Moreover, TaPUB4 facilitated stomatal response to elevated CO 2 emission rates under ABA conditions. TaPUB4-overexpressing Arabidopsis, on the other hand, was more resistant to drought stress in plant development, demonstrating that TaPUB4 positively regulates drought-mediated control of plant growth. Moreover, the ectopic expression of the TaPUB4 gene was significant influential in drought sensitive metrics including survival rate, chlorophyll content, water loss, proline content and the expression of drought stress-response genes. Collectively, our results demonstrate that TaPUB4 may regulate drought stress response and ABA conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

Background: Lettuce is one of the most extensively farmed vegetables in the world, and it prefers cool growing conditions. High temperatures promote premature bolt formation, reducing quality and yield. The gibberellic acid-stimulated Arabidopsis (GASA) family genes play critical roles in plant growth, development, and stress responses. However, the biological functions of GASA proteins in lettuce have yet to be thoroughly investigated., Results: Using genome-wide analysis, 20 GASAs were identified in lettuce including, three groups of LsGASA proteins based on the phylogenetic analysis. Except for one, all GASA proteins included a conserved GASA domain with 12 cysteine residues. Cis-element analysis showed that LsGASAs were closely associated with light, phytohormones, and stress resistance. Five segmental and three tandem duplication events were observed in the LsGASA family based on duplication analysis. GASA synteny analysis among lettuce, Arabidopsis, tobacco, and rice revealed that LsGASA5 is highly collinear with all species. Six of the 20 LsGASA showed increased expression patterns at specific time points in the shoot apical meristem when subjected to heat stress. According to gene expression analysis, the majority of GASA were highly expressed in flowers compared to other organs, and six GASA exhibited highly increased expression levels in response to NaCl, abscisic acid, and gibberellin treatment. Furthermore, LsGASA proteins are predominantly found in the plasma membrane and/or the cytosol., Conclusions: This study provides a comprehensive characterization of LsGASA genes for their diversity and biological functions. Moreover, our results will be useful for further studies on the function of lettuce GASA in abiotic stress- and heat-induced bolting signaling., (© 2023. The Author(s).)

Effective surveillance during smart cities' sustainable development allows their cleanliness to be maintained by reusing waste to produce renewable energy. This study quantifies the biodegradable waste generated in specific regions of several cities and presents ways to convert it into renewable energy. This energy can then be used to partially support regional energy demands. This study explores ways of reducing carbon emissions for biodegradable waste collection processes in regional centers, ultimately sending the biodegradable waste to the energy conversion center. The smart production system allows for the flexible production and autonomation of rates of conversion; green technology depends on each regional center's research management, which is a decision variable for reducing carbon emissions. The major contribution of this study is to consider an energy supply chain management with flexibility of energy conversion under the reduction of carbon emissions, which leads to a sustainable ESCM with the global maximum profit. This study uses mathematical modeling to decrease biodegradable waste with conversion of energy through a classical optimization technique. The solution to this mathematical model yielded significant results, providing insight into waste reduction, reduced carbon emissions and the conversion of biodegradable waste to energy. The model is examined using numerical experiments, and its conclusion supports the model with the fundamental assumptions. Results of sensitivity analysis provide insight into the reduction and re-utilization of wastes, carbon emission reduction, and the benefits of using renewable energy.

Plant U-box E3 ligases (PUBs) are important regulators of responses to various abiotic stress conditions. In this study, we found that wheat (Triticum aestivum. L) PUBs TaPUB2 and TaPUB3 enhanced abscisic acid (ABA) responses and salt tolerance in Arabidopsis. We generated transgenic Arabidopsis lines overexpressing TaPUB2 and TaPUB3 and performed various plant physiological experiments. Overexpression of TaPUB2 and TaPUB3 increased tolerance to salinity stress in an ABA-dependent manner in transgenic plants, as evidenced by germination and survival rates, root length, stomatal aperture regulation, membrane peroxidation, photosynthetic activities, reactive oxygen species scavenging activities and expression of various ABA and salinity stress-related genes. These results demonstrate the functions of PUBs under ABA and salinity stress conditions and provide valuable information for the development of salinity stress-tolerant crop species., (© 2022 Federation of European Biochemical Societies.)

This study investigated the correlation between spontaneous pneumothorax (SP) and meteorological factors during different seasons. Patients who visited emergency rooms (ERs) in large cities in Korea and were discharged with SP from 2014 to 2016 were included in this study. Data on temperature, air pressure, and wind speed for each region were collected to obtain each factor's daily maximum, minimum, average, and changes. Days with more than 1 case of SP per million were referred to as pneumothorax days (PD) and those with less than 1 case of SP per million were referred to as non-pneumothorax days (NPD). The environmental factors were assessed on the same day (Day 0), 1 day prior (Day-1), and 2 days prior (Day-2) to PD and NPD per season. A total of 17,846 patients were included in this study. During winter, 4080 patients with SP visited the ERs of large cities with low population densities. The maximum temperature (0.16°C vs 0.76°C, 0.04°C vs 0.87°C, and 0.09°C vs 0.91°C), change in temperature (0.24°C vs 0.90°C, 0.38°C vs 0.81°C, and 0.41°C vs 0.83°C), average atmospheric pressure (0.16 vs 0.52 hPa, 0.25 vs 0.42 hPa, 0.34 vs 0.40 hPa), and maximum atmospheric pressure (0.15 vs 0.53 hPa, 0.28 vs 0.49 hPa, 0.33 vs 0.71 hPa) were greater for Day 0, Day-1, and Day-2, respectively, in PD than in NPD. Meanwhile, the average (0.31 vs 0.48 m/s, 0.28 vs 0.46 m/s, 0.20 vs 0.40 m/s), minimum (0.20 vs 0.31 m/s, 0.18 vs 0.25 m/s, 0.16 vs 0.25 m/s), and maximum (0.44 vs 0.67 m/s, 0.36 vs 0.71 m/s, 0.26 vs 0.58 m/s) wind speeds were slower, and the changes in wind speed (0.44 vs 0.67 m/s, 0.36 vs 0.71 m/s, 0.16 vs 0.25 m/s) were lower for all 3 days in PD than in NPD. High average and change in temperature, slow and unchanging wind speed, and high average and maximum atmospheric pressure were associated with SP. Since many findings of this study were contradictory to previous studies, it is assumed that the interaction of various factors affects SP., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2022 the Author(s). Published by Wolters Kluwer Health, Inc.)

Rationale: Since tobacco contains numerous harmful substances, besides nicotine, which is addictive, smoking cessation products and tobacco alternatives, such as electronic (e-) cigarettes, nicotine chewing gums, and patches, are being widely used. Nicotine gums are consumed orally. The nicotine from the gum is absorbed at a slower rate than that from e-cigarettes, and the former remains in the bloodstream for a longer period. In addition, the maximum number of daily doses is high, and it can be purchased without a doctor's prescription in many countries., Patient Concerns: A 29-year-old male patient consumed 5 2-mg nicotine gums at a time, twice a day, for 4 days (total amount: 70 mg). However, he visited the emergency unit with the chief complaint of involuntary limb movements after consuming an additional 15 gums 3 hour before the visit. At admission, his consciousness was clear, although 2 hour later, he experienced sudden loss of consciousness with worsening hypoxia and respiratory acidosis., Diagnosis: The patient's vital signs were stable at the time of admission, and blood test results showed no specific findings other than a white blood cell count of 14,800/µL, lactate level of 6.4 mmol/L, and prolactin level of 119.02 ng/mL. In addition, chest radiography and head computed tomography scans showed no acute phase abnormalities. Two hours later, he experienced loss of consciousness and respiratory failure, and the results of blood tests performed at this time showed that his blood cotinine level was 3491 ng/mL., Interventions: Supportive treatment, including endotracheal intubation followed by mechanical ventilation, was provided., Outcome: The patient's vital signs stabilized 3 days after treatment, and his consciousness and respiratory status had improved; therefore, mechanical ventilation was stopped. His condition was stable for the next 2 days, and he was discharged on the fifth day., Lessons: Acute respiratory exacerbation due to nicotine poisoning (from levels exceeding the lethal dose of 30-60 mg) was observed, although the gums were consumed over several days. Patients with nicotine poisoning may show acute respiratory failure and should be monitored carefully. Further studies are required to determine the toxic effects of nicotine replacement therapies., Competing Interests: The authors have no funding and conflicts of interest to disclose., (Copyright © 2022 the Author(s). Published by Wolters Kluwer Health, Inc.)

Although many concerns have been raised on increased self-harm or suicide attempts since the emergence of the coronavirus disease 2019 (COVID-19) pandemic, the numbers of studies reported no consistent increase. This study aimed to analyze the data on the request for emergency medical service (EMS) in Daegu Metropolitan City in Korea to investigate the effects of the COVID-19 pandemic on the incidence and types of suicidal patients. Data of 4480 cases requesting EMS related to self-harm or suicide 1 year before and after the COVID-19 pandemic were retrospectively comparatively analyzed (February 19, 2019-February 18, 2021). The number of EMS requests for self-harm and suicide increased after the pandemic compared to that before the pandemic (daily mean request 5.83 [±2.597] vs 6.43 [±2.918]). In particular, the number of female patients increased per day on average (2.61 [±1.717] vs 3.17 [±1.893]). With respect to the reasons for the request, committed self-harm and attempts to commit self-harm increased, whereas the presumption against suicide decreased. With respect to consciousness levels, the number of alert patients increased, whereas the number of transport cases decreased. For the method of the attempt, hanging and carbon monoxide/gas poisoning decreased, whereas jumping from a height and drowning increased. The number of patients with psychiatric history and those with other chronic illnesses increased. In multivariate regression analysis, women (OR 1.227, 95% CI = 1.072-1.405, P = .003), patients with psychiatric diseases (OR 1.223, 95% CI = 1.031-1.450, P = .021), patients with other chronic illnesses (OR 1.510, 95% CI = 1.127-2.023, P = .006), and CO or gas poisoning (not attempted) (OR 1.349, 95% CI = 1.038-1.753, P = .025) showed statistically significant differences. Among the request for EMS, requests for committed self-harm and attempts to commit suicide increased. Medical support and measures for mental health and emergency medical systems should be established for female patients and patients with psychiatric or other chronic diseases., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2022 the Author(s). Published by Wolters Kluwer Health, Inc.)

Cochlear afferent nerve fibers (ANF) are the first neurons in the ascending auditory pathway. We investigated the low-voltage activating K + channels expressed in ANF dendrites using isolated rat cochlear segments. Whole cell patch clamp recordings were made from the dendritic terminals of ANFs. Outward currents activating at membrane potentials as low as -64 mV were observed in all dendrites studied. These currents were inhibited by 4-aminopyridine (4-AP), a blocker known to preferentially inhibit low-voltage activating K + currents (I KL ) in CNS auditory neurons and spiral ganglion neurons. When the dendritic I KL was blocked by 4-AP, the EPSP decay time was significantly prolonged, suggesting that dendritic I KL speeds up the decay of EPSPs and likely modulates action potentials of ANFs. To reveal molecular subtype of dendritic I KL , α-dendrotoxin (α-DTX), a selective inhibitor for K v 1.1, K v 1.2, and K v 1.6 containing channels, was tested. α-DTX inhibited 23±9% of dendritic I KL . To identify the α-DTXsensitive and α-DTX-insensitive components of I KL , immunofluorescence labeling was performed. Strong K v 1.1- and K v 1.2-immunoreactivity was found at unmyelinated dendritic segments, nodes of Ranvier, and cell bodies of most ANFs. A small fraction of ANF dendrites showed K v 7.2- immunoreactivity. These data suggest that dendritic I KL is conducted through K v 1.1and K v 1.2 channels, with a minor contribution from K v 7.2 and other as yet unidentified channels.

Mott threshold switching, which is observed in quantum materials featuring an electrically fired insulator-to-metal transition, calls for delicate control of the percolative dynamics of electrically switchable domains on a nanoscale. Here, we demonstrate that embedded metallic nanoparticles (NP) dramatically promote metastability of switchable metallic domains in single-crystal-like VO 2 Mott switches. Using a model system of Pt-NP-VO 2 single-crystal-like films, interestingly, the embedded Pt NPs provide 33.3 times longer 'memory' of previous threshold metallic conduction by serving as pre-formed 'stepping-stones' in the switchable VO 2 matrix by consecutive electical pulse measurement; persistent memory of previous firing during the application of sub-threshold pulses was achieved on a six orders of magnitude longer timescale than the single-pulse recovery time of the insulating resistance in Pt-NP-VO 2 Mott switches. This discovery offers a fundamental strategy to exploit the geometric evolution of switchable domains in electrically fired transition and potential applications for non-Boolean computing using quantum materials., (© 2022. The Author(s).)

Competitive college admissions and academic pressure have continuously increased the psychopathological burden of Korean high school students. Seeking help is one of the primary means of managing mental health, and more attention is required. This study aimed to explore the psychopathological and cognitive factors related to the help-seeking intentions of Korean high school students. This cross-sectional study was conducted between July and August 2020 using the General Help-Seeking Questionnaire, Symptom Checklist-90-R, and Mental Health Literacy Scale. Four hundred and twenty-one Korean high school students (275 males, 146 females; average age 17.44 years [standard deviation = 0.651]) completed self-report questionnaires. We performed analysis of variance, Spearman's correlation analysis, and stepwise regression analysis to explore the factors related to help-seeking intentions. The final model showed an explanatory power of 23.6% for the overall variance in help-seeking intentions. Somatization (β = -0.200; P = .001) and hostility (β = -0.203; P = .001) had a negative effect on help-seeking intentions. Further, knowledge of where to seek information (β = 0.230; P < .001) and attitudes promoting recognition and help-seeking behavior (β = 0.095; P = .030) had a positive effect. Students responded to society's negative awareness of mental illness by converting psychopathology into socially acceptable symptoms. Educational support can improve mental health literacy. This study is expected to help improve mental illness awareness and increase adolescents' access to public services., Competing Interests: The authors have no funding and conflicts of interest to disclose., (Copyright © 2022 the Author(s). Published by Wolters Kluwer Health, Inc.)

Background: Biphasic calcium phosphate (BCP) is the most frequently used synthetic bone substitutes, which comprises a combination of hydroxyapatite (HA) and beta-tricalcium phosphate (b-TCP). Thanks to the recent advances in digital dentistry and three-dimensional (3D) printing technology, synthetic block bone substitutes can be customized to fit individual defect morphologies. The diameter of the pores can influence the rate of bone formation and material resorption. The aim of this study was to compare three-dimensionally printed biphasic calcium phosphate (BCP) block bone substitutes with different pore diameters (0.8-, 1.0-, and 1.2- mm) for use in the regeneration of rabbit calvarial defects., Methods: Four circular defects were formed on the calvaria of ten rabbits. Each defect was randomly allocated to one of the following study groups: (i) control group, (ii) 0.8-mm group, (iii) 1.0-mm group, and (iv) 1.2-mm group. All specimens were postoperatively harvested at 2 and 8 weeks, and radiographic and histomorphometric analyses were performed on the samples., Results: Histologically, the BCP blocks remained unresorbed up to 8 weeks, and new bone formation occurred within the porous structures of the blocks. After the short healing period of 2 weeks, histomorphometric analysis indicated that new bone formation was significantly greater in the BCP groups compared with the control (p < 0.05). However, there were no significant differences between the groups with different pore diameters (p > 0.05). At 8 weeks, only the 1.0-mm group (3.42 ± 0.48 mm 2 , mean ± standard deviation) presented a significantly larger area of new bone compared with the control (2.26 ± 0.59 mm 2 ) (p < 0.05). Among the BCP groups, the 1.0- and 1.2-mm groups exhibited significantly larger areas of new bone compared with the 0.8-mm group (3.42 ± 0.48 and 3.04 ± 0.66 vs 1.60 ± 0.70 mm 2 , respectively)., Conclusions: Within the limitations of this study, the BCP block bone substitutes can be applied to bone defects for successful bone regeneration. Future studies should investigate more-challenging defect configurations prior to considering clinical applications., (© 2022. The Author(s).)

Plants accumulate key metabolites as a response of biotic/abiotic stress conditions. In seed coats, anthocyanins, carotenoids, and chlorophylls can be found. They have been associated as important antioxidants that affect germination. In wheat, anthocyanins can impart the seed coat color which have been recognized as health-promoting nutrients. Transcription factors act as master regulators of cellular processes. Transcription complexes such as MYB-bHLH-WD40 (MBW) regulate the expression of multiple target genes in various plant species. In this study, the spatiotemporal accumulation of seed coat pigments in different developmental stages (10, 20, 30, and 40 days after pollination) was analyzed using cryo-cuts. Moreover, the accumulation of phenolic, anthocyanin, and chlorophyll contents was quantified, and the expression of flavonoid biosynthetic genes was evaluated. Finally, transcriptome analysis was performed to analyze putative MYB genes related to seed coat color, followed by further characterization of putative genes. TaTCL2 , an MYB gene, was cloned and sequenced. It was determined that TaTCL2 contains a SANT domain, which is often present in proteins participating in the response to anthocyanin accumulation. Moreover, TaTCL2 transcript levels were shown to be influenced by anthocyanin accumulation during grain development. Interaction network analysis showed interactions with GL2 (HD-ZIP IV), EGL3 (bHLH), and TTG1 (WD40). The findings of this study elucidate the mechanisms underlying color formation in Triticum aestivum L. seed coats., Competing Interests: Declaration of Conflicting Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2022.)

Wheat ( Triticum aestivum ) has diverse uses in the food industry, and different cultivars have unique properties; therefore, it is important to select the optimal cultivar for the intended end use. Here, to establish an identification system for Korean wheat cultivars, we obtained the complete plastome sequences of seven major Korean cultivars. Additionally, the open access database CerealsDB was queried to discover single-copy genomic single-nucleotide polymorphisms (SNPs) in the hexaploid wheat genome. Ten SNPs were developed into allele-specific PCR (ASP) markers, and eight of the SNPs used for ASP markers were converted into TaqMan high-throughput genotyping markers. Phylogenetic analysis using SNP genotypes revealed the genetic diversity and relationships among 137 wheat lines from around the world, including 35 Korean cultivars. This research thus presents a high-throughput authentication system for Korean wheat cultivars that may promote food industry uses of Korean wheat., Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01043-w., Competing Interests: Conflict of interestThe researchers claim no conflicts of interest., (© The Korean Society of Food Science and Technology 2022.)

In wheat (Triticum aestivum L.), the floret development stage is an important step in determining grain yield per spike; however, the molecular mechanisms underlying floret development remain unclear. In this study, we elucidated the role of TaF-box2, a member of the F-box-containing E3 ubiquitin protein ligases, which is involved in floret development and anthesis of wheat. TaF-box2 was transiently expressed in the plasma membrane and cytoplasm of both tobacco and wheat. We also found that the SCF F-box2 (Skp1-Cul1-Rbx1-TaF-box2) ubiquitin ligase complex mediated self-ubiquitination activity. Transgenic Arabidopsis plants that constitutively overexpressed TaF-box2 showed markedly greater hypocotyl and root length than wild-type plants, and produced early flowering phenotypes. Flowering-related genes were significantly upregulated in TaF-box2-overexpressing Arabidopsis plants. Further protein interaction analyses such as yeast two-hybrid, in vitro pull-down, and bimolecular fluorescence complementation assays confirmed that TaF-box2 physically interacted with TaCYCL1 (Triticum aestivum cyclin-L1-1). Ubiquitination and degradation assays demonstrated that TaCYCL1 was ubiquitinated by SCF F-box2 and degraded through the 26S proteasome complex. The physiological functions of the TaF-box2 protein remain unclear; however, we discuss several potential routes of involvement in various physiological mechanisms which counteract flowering in transgenic Arabidopsis plants., (© 2022 Scandinavian Plant Physiology Society.)

Plant U-box E3 ubiquitin ligase (PUB) is involved in various environmental stress conditions. However, the molecular mechanism of U-box proteins in response to abiotic stress in wheat remains unknown. In this study, two U-box E3 ligase genes ( TaPUB2 and TaPUB3 ), which are highly expressed in response to adverse abiotic stresses, were isolated from common wheat, and their cellular functions were characterized under drought stress. Transient expression assay revealed that TaPUB2 was localized in the cytoplasm and Golgi apparatus, whereas TaPUB3 was expressed only in the Golgi apparatus in wheat protoplasts. Additionally, TaPUB2 and TaPUB3 underwent self-ubiquitination. Moreover, TaPUB2/TaPUB3 heterodimer was identified in yeast and the cytoplasm of wheat protoplasts using a pull-down assay and bimolecular fluorescence complementation analysis. Heterogeneous overexpression of TaPUB2 and TaPUB3 conferred tolerance to drought stress. Taken together, these results implied that the heterodimeric form of U-box E3 ubiquitin ligases (TaPUB2/TaPUB3) responded to abiotic stress and roles as a positive regulator of drought stress tolerance.

The study aimed to decipher the impact of multiple drought stress on wheat. To that effect, Geumgangmil, PL 337 (1AL.1RS), PL 371 (1BL.1RS), and PL 257 (1DL.1RS) seedlings were subjected to four treatments: G1 (control), G2 (stressed thrice with rewatering), G3 (stressed twice with rewatering), and G4 (single stressful event). The findings provided a comprehensive framework of drought-hardening effect at physiological, biochemical, and gene expression levels of drought-stressed wheat genotypes. The treatments resulted in differentially higher levels of malondialdehyde (MDA), hydrogen peroxide (H 2 O 2 ), soluble sugar, and proline accumulation, and reduced relative water content (RWC) in wheat plants. Photosynthetic pigment (chlorophyll and carotenoid) levels, the membrane stability index (MSI), and shoot biomass decreased dramatically and differently across genotypes, particularly in G3 and G4 compared to G2. The activity of antioxidant enzymes [ascorbate peroxidase (APX), superoxide dismutase (SOD), and catalase (CAT)] increased with the duration and severity of drought treatment. Furthermore, the relative expression of DREB , LEA , HSP , P5CS , SOD1 , CAT1 , APX1 , RBCL , and CCD1 genes was higher in G2 than in other treatments. Drought hardening increased drought tolerance and adaptability in plants under G2 by enhancing growth and activating defensive mechanisms at the physio-biochemical and molecular levels. The findings of the study indicated that early drought stress exposure-induced acclimation (hardening), which enhanced tolerance to subsequent drought stress in wheat seedlings. The findings of this study will be useful in initiating a breeding program to develop wheat cultivars with improved drought tolerance., Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-021-02991-6., Competing Interests: Conflict of interestNo potential conflict of interest is reported by the authors., (© King Abdulaziz City for Science and Technology 2021.)

Background: Bread wheat (Triticum aestivum L.) is one of the most widely consumed cereal crops, but its complex genome makes it difficult to investigate the genetic effect on important agronomic traits. Genome-wide association (GWA) analysis is a useful method to identify genetic loci controlling complex phenotypic traits. With the RNA-sequencing based gene expression analysis, putative candidate genes governing important agronomic trait can be suggested and also molecular markers can be developed., Results: We observed major quantitative agronomic traits of wheat; the winter survival rate (WSR), days to heading (DTH), days to maturity (DTM), stem length (SL), spike length (SPL), awn length (AL), liter weight (LW), thousand kernel weight (TKW), and the number of seeds per spike (SPS), of 287 wheat accessions from diverse country origins. A significant correlation was observed between the observed traits, and the wheat genotypes were divided into three subpopulations according to the population structure analysis. The best linear unbiased prediction (BLUP) values of the genotypic effect for each trait under different environments were predicted, and these were used for GWA analysis based on a mixed linear model (MLM). A total of 254 highly significant marker-trait associations (MTAs) were identified, and 28 candidate genes closely located to the significant markers were predicted by searching the wheat reference genome and RNAseq data. Further, it was shown that the phenotypic traits were significantly affected by the accumulation of favorable or unfavorable alleles., Conclusions: From this study, newly identified MTA and putative agronomically useful genes will help to study molecular mechanism of each phenotypic trait. Further, the agronomically favorable alleles found in this study can be used to develop wheats with superior agronomic traits., (© 2021. The Author(s).)

F-box proteins are substrate recognition components of the Skp1-Cullin-F-box (SCF) complex, which performs many important biological functions including the degradation of numerous proteins via the ubiquitin-26S proteasome system. In this study, we isolated the gene encoding the F-box/LRR-repeat (FBXL) protein from wheat ( Triticum aestivum L.) seedlings and validated that the TaFBXL protein is a component of the SCF complex. Yeast two-hybrid assays revealed that TaFBXL interacts with the wheat glycosylphosphatidylinositol-anchored protein (TaGPI-AP). The green fluorescent protein (GFP) fusion protein of TaFBXL was detected in the nucleus and plasma membrane, whereas that of TaGPI-AP was observed in the cytosol and probably also plasma membrane. yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays revealed that TaFBXL specifically interacts with TaGPI-AP in the nucleus and plasma membrane, and TaGPI-AP is targeted by TaFBXL for degradation via the 26S proteasome system. In addition, TaFBXL and TaGPI-AP showed antagonistic expression patterns upon treatment with indole-3-acetic acid (IAA), and the level of TaGPI-AP was higher in tobacco leaves treated with both MG132 (proteasome inhibitor) and IAA than in leaves treated with either MG132 or IAA. Taken together, our data suggest that TaFBXL regulates the TaGPI-AP protein level in response to exogenous auxin application.

Abstract: Emergency departments (EDs) are on the frontline of the coronavirus disease (COVID-19) outbreak. To resolve the abrupt overloading of COVID-19-suspected patients in a community, each ED needs to respond in various ways. In our hospital, we increased the isolation beds through temporary remodeling and by performing in-hospital COVID-19 polymerase chain reaction testing rather than outsourcing them. The aim of this study was to verify the effects of our response to the newly developed viral outbreak.The medical records of patients who presented to an ED were analyzed retrospectively. We divided the study period into 3: pre-COVID-19, transition period of response (the period before fully implementing the response measures), and post-response (the period after complete response). We compared the parameters of the National Emergency Department Information System and information about isolation and COVID-19.The number of daily ED patients was 86.8 ± 15.4 in the pre-COVID-19, 36.3 ± 13.6 in the transition period, and 67.2 ± 10.0 in the post-response period (P < .001). The lengths of stay in the ED were significantly higher in transition period than in the other periods [pre-COVID-19 period, 219.0 (121.0-378.0) min; transition period, 301 (150.0-766.5) min; post-response period, 281.0 (114.0-575.0) min; P < .001]. The ratios of use of an isolation room and fever (≥37.5°C) were highest in the post-response period [use of isolation room: pre-COVID-19 period, 0.6 (0.7%); transition period, 1.2 (3.3%); post-response period, 16.1 (24.0%); P < .001; fever: pre-COVID-19 period, 14.8(17.3%); transition period, 6.8 (19.1%); post-response period, 14.5 (21.9%), P < .001].During an outbreak of a novel infectious disease, increasing the number of isolation rooms in the ED and applying a rapid confirmation test would enable the accommodation of more suspected patients, which could help reduce the risk posed to the community and thus prevent strain on the local emergency medical system., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2021 the Author(s). Published by Wolters Kluwer Health, Inc.)

Late stages of floret development, such as booting, heading, and anthesis stages, are important steps for determining grain setting and for filling in wheat. Herein, we report the molecular function of Triticum aestivum ELF7 encoding RNA polymerase II-associated factor 1 (PAF1), which may act as a negative regulator in floret development and anthesis stages. Among the six TaELF7-like genes isolated from wheat, TaELF7 like1-A and TaELF7 like2-B showed contrasting expression levels during the late stage of floret development stages, with observation of decreased expression level of TaELF7 like1-A compared to that of TaELF7 like2-B. The full-length TaELF7 like1-A has a 1038-bp open reading frame that contains a proline-rich domain in the N-terminal region and a nuclear localization signal domain in the C-terminal region. TaELF7 like1-A was found to be localized in the nucleus in both tobacco and wheat. Direct interaction of TaELF7 with the RING-type E3 ligase TaHUB2 was confirmed using a yeast two-hybrid system, an in vitro pull-down assay, and a bimolecular fluorescence complementation assay. The flowering time was delayed in TaELF7-overexpressing plants compared to that in the control plants. Expression levels of few floral repressor genes were markedly increased in TaELF7-overexpressing Arabidopsis plants., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Background: Flavonoids can protect plants against extreme temperatures and ROS due to their antioxidant activities. We found that deep-purple seed coat color was controlled by two gene interaction (12:3:1) from the cross between yellow and deep-purple seed coat colored inbreds. F 2:3 seeds were grouped in 3 by seed coat color and germinated under chilling (4 °C) and non-acclimated conditions (18 °C) for a week, followed by normal conditions (18 °C) for three weeks and a subsequent chilling stress (4 °C) induction. We analyzed mean daily germination in each group. Additionally, to study the acclimation in relationship to the different seed coat colors on the germination ability and seedling performances under the cold temperatures, we measured the chlorophyll content, ROS scavenging activity, and expression levels of genes involved in ROS scavenging, flavonoid biosynthetic pathway, and cold response in seedlings., Results: The results of seed color segregation between yellow and deep purple suggested a two-gene model. In the germination study, normal environmental conditions induced the germination of yellow-seed, while under chilling conditions, the germination ratio of deep purple-seed was higher than that of yellow-colored seeds. We also found that the darker seed coat colors were highly responsive to cold acclimation based on the ROS scavenging enzymes activity and gene expression of ROS scavenging enzymes, flavonoid biosynthetic pathway and cold responsive genes., Conclusions: We suggest that deep purple colored seed might be in a state of innate pre-acquired stress response state under normal conditions to counteract stresses in a more effective way. Whereas, after the acclimation, another stress should enhance the cold genes expression response, which might result in a more efficient chilling stress response in deep purple seed seedlings. Low temperature has a large impact on the yield of crops. Thus, understanding the benefit of seed coat color response to chilling stress and the identification of limiting factors are useful for developing breeding strategies in order to improve the yield of wheat under chilling stress.

High-temperature stress during the grain filling stage has a deleterious effect on grain yield and end-use quality. Plants undergo various transcriptional events of protein complexity as defensive responses to various stressors. The " Keumgang " wheat cultivar was subjected to high-temperature stress for 6 and 10 days beginning 9 days after anthesis, then two-dimensional gel electrophoresis (2DE) and peptide analyses were performed. Spots showing decreased contents in stressed plants were shown to have strong similarities with a high-molecular glutenin gene, TraesCS1D02G317301 ( TaHMW1D ). QRT-PCR results confirmed that TaHMW1D was expressed in its full form and in the form of four different transcript variants. These events always occurred between repetitive regions at specific deletion sites (5'-CAA (Glutamine) GG/TG (Glycine) or (Valine)-3', 5'-GGG (Glycine) CAA (Glutamine) -3') in an exonic region. Heat stress led to a significant increase in the expression of the transcript variants. This was most evident in the distal parts of the spike. Considering the importance of high-molecular weight glutenin subunits of seed storage proteins, stressed plants might choose shorter polypeptides while retaining glutenin function, thus maintaining the expression of glutenin motifs and conserved sites.

U-box E3 ligase genes play specific roles in protein degradation by post-translational modification in plant signaling pathways, developmental stages, and stress responses; however, little is known about U-box E3 genes in wheat. We identified 213 U-box E3 genes in wheat based on U-box and other functional domains in their genome sequences. The U-box E3 genes were distributed among 21 chromosomes and most showed high sequence homology with homoeologous U-box E3 genes. Synteny analysis of wheat U-box E3 genes was conducted with other plant species such as Brachypodium distachyon , barley, rice, Triricum uratu, and Aegilops tauschii . A total of 209 RNA-seq samples representing 22 tissue types, from grain, root, leaf, and spike samples across multiple time points, were analyzed for clustering of U-box E3 gene expression during developmental stages, and the genes responded differently in various tissues and developmental stages. In addition, expression analysis of U-box E3 genes under abiotic stress, including drought, heat, and both heat and drought, and cold conditions, was conducted to provide information on U-box E3 gene expression under specific stress conditions. This analysis of U-box E3 genes could provide valuable information to elucidate biological functions for a better understanding of U-box E3 genes in wheat.

Key Message: We identified a RING-type E3 ligase (TaBAH1) protein in winter wheat that targets TaSAHH1 for degradation and might be involved in primordia development by regulating targeted protein degradation. Grain yield per spike in wheat (Triticum aestivum), is mainly determined prior to flowering during mature primordia development; however, the genes involved in primordia development have yet to be characterized. In this study, we demonstrated that, after vernalization for 50 days at 4 °C, there was a rapid acceleration in primordia development to the mature stages in the winter wheat cultivars Keumgang and Yeongkwang compared with the Chinese Spring cultivar. Although Yeongkwang flowers later than Keumgang under normal condition, it has the same heading time and reaches the WS9 stage of floral development after vernalization for 50 days. Using RNA sequencing, we identified candidate genes associated with primordia development in cvs. Keumgang and Yeongkwang, that are differentially expressed during wheat reproductive stages. Among these, the RING-type E3 ligase TaBAH1 (TraesCS5B01G373000) was transcriptionally upregulated between the double-ridge (WS2.5) stage and later stages of floret primordia development (WS10) after vernalization. Transient expression analysis indicated that TaBAH1 was localized to the plasma membrane and nucleus and was characterized by self-ubiquitination activity. Furthermore, we found that TaBAH1 interacts with TaSAHH1 to mediate its polyubiquitination and degradation through a 26S proteasomal pathway. Collectively, the findings of this study indicate that TaBAH1 might play a prominent role in post-vernalization floret primordia development.

Background: Cardiopulmonary resuscitation (CPR) performance depends on individual ability and training. Well-trained or professional rescuers can maintain high-quality CPR for longer than laypeople. This study aimed to examine the effects of reducing resting intervals on CPR performance, physiological parameters, and hemodynamic parameters during prolonged CPR in well-trained providers., Methods: The study enrolled 90 volunteers from the paramedic students of our institution. They were randomly divided into 3 groups: 2 minutes, 1 minute 45 seconds, and 1 minute 30 seconds rest groups. Each participant performed 5 cycles of chest compression only CPR (2 min/cycle) with different resting intervals according to grouping. CPR quality, physiological variations, and hemodynamic variations were measured for each cycle and compared across the groups., Results: Of the 90 volunteers, 79 well-trained providers were finally included. The variation of the average chest compression depth across the 5 cycles showed significant differences between the 3 groups: from cycle 1 to 2: 1.2 (3.1) mm, -0.8 (2.0) mm, and -2.0 (3.0) mm in the 2 minutes, 1 minute 45 seconds, and 1 minute 30 seconds groups, respectively (P < .001); from cycle 1 to 3: 0.0 (3.0) mm, -0.7 (3.2) mm, and -2.6 (3.9) mm, respectively (P = .030). However, all 3 groups maintained the recommended rate and chest compression depth for all 5 cycles. Physiological and hemodynamic parameters showed no significant differences between the groups., Conclusions: Well-trained providers were able to maintain high-quality CPR despite reducing rest intervals. Adjusting the rest interval may help maintain overall CPR quality in special situations or where layperson rescuers are involved., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2021 the Author(s). Published by Wolters Kluwer Health, Inc.)

Abscisic acid-, stress-, and ripening-induced (ASR) proteins play an important role in protecting plants against adverse environmental conditions. Here, we identified 24 ASR genes in the wheat genome and analyzed their characteristics. Among these, five ASR genes highly induced by abscisic acid (ABA) and polyethylene glycol were cloned and further characterized. The TaASR genes were expressed in response to different abiotic stresses and ABA and were found to be localized in the nucleus and plasma membrane of transformed tobacco cells. Brachypodium distachyon transgenic plants overexpressing TaASR2D showed enhanced drought tolerance by regulating leaf transpiration. The expression levels of stress-related and ABA-responsive genes were higher in transgenic plants than in wild-type plants under drought stress conditions. Moreover, overexpression of TaASR2D increased the levels of both endogenous ABA and hydrogen peroxide in response to drought stress, and these plants showed hypersensitivity to exogenous ABA at the germination stage. Furthermore, plants overexpressing TaASR2D showed increased stomatal closure. Further analysis revealed that TaASR2D interacts with ABA biosynthesis and stress-related proteins in yeast and tobacco plants. Collectively, these findings indicate that TaASR2D plays an important role in the response of plants to drought stress by regulating the ABA biosynthesis pathway and redox homeostasis system., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)