Research最新文献

{"title":"Microrobotic Swarms for Cancer Therapy.","authors":"Leiming Xie, Jinbo Liu, Zhen Yang, Hui Chen, Yibin Wang, Xingzhou Du, Yongping Fu, Peng Song, Jiangfan Yu","doi":"10.34133/research.0686","DOIUrl":"https://doi.org/10.34133/research.0686","url":null,"abstract":"<p><p>Microrobotic swarms hold great promise for the revolution of cancer treatment. The coordination of miniaturized microrobots offers a unique approach to treating cancers at the cellular level with enhanced delivery efficiency and environmental adaptability. Prior studies have summarized the design, functionalization, and biomedical applications of microrobotic swarms. The strategies for actuation and motion control of swarms have also been introduced. In this review, we first give a detailed introduction to microrobot swarming. We then explore the design of microrobots and microrobotic swarms specifically engineered for cancer therapy, with a focus on tumor targeting, infiltration, and therapeutic efficacy. Moreover, the latest developments in active delivery methods and imaging techniques that enhance the precision of these systems are discussed. Finally, we categorize and analyze the various cancer therapies facilitated by functional microrobotic swarms, highlighting their potential to revolutionize treatment strategies for different cancer types.</p>","PeriodicalId":21120,"journal":{"name":"Research","volume":"8 ","pages":"0686"},"PeriodicalIF":11.0,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12038165/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144009716","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Discovery of a Novel Selective PAK1/HDAC6/HDAC10 Inhibitor ZMF-25 that Induces Mitochondrial Metabolic Breakdown and Autophagy-Related Cell Death in Triple-Negative Breast Cancer.","authors":"Jin Zhang, Xiaoling Cheng, Gang Chen, Xiya Chen, Xi Zhao, Weiji Chen, Wei Du, Zhendan He, Xiaojun Yao, Bo Han, Dahong Yao","doi":"10.34133/research.0670","DOIUrl":"https://doi.org/10.34133/research.0670","url":null,"abstract":"<p><p>Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subtype, and addressing its intrinsic heterogeneity has emerged as a valuable avenue for novel clinical treatment strategy. Here, we put forward an innovative strategy for TNBC treatment by simultaneously suppressing both p21-activated kinase 1 (PAK1) and histone deacetylase (HDAC) class IIb (HDAC6/10). A series of pyrido [2,3-d]pyrimidin-7(8<i>H</i>)-one moiety derivatives was successfully designed and synthesized to target PAK1/HDAC6/HDAC10 by utilizing structure-based screening and pharmacophore integration. ZMF-25 demonstrates marked inhibitory activity against PAK1, HDAC6, and HDAC10 with respective IC₅₀ values of 33, 64, and 41 nM, remarkable selectivity over HDACs and PAKs, as well as prominent antiproliferative efficiency in MDA-MB-231 cells. Additionally, ZMF-25 effectively suppresses TNBC proliferation and migration by inhibiting PAK1/HDAC6/HDAC10. Moreover, it was found to impair glycolysis and trigger reactive oxygen species generation, resulting in autophagy-related cell death by inhibiting the AKT/mTOR/ULK1 signaling. Furthermore, ZMF-25 exhibits remarkable therapeutic potential with no obvious toxicity in vivo and good pharmacokinetics. In summary, these observations indicate that ZMF-25 is a novel and potent triple-targeting PAK1/HDAC6/HDAC10 inhibitor, which is expected to provide a novel and effective strategy for TNBC treatment.</p>","PeriodicalId":21120,"journal":{"name":"Research","volume":"8 ","pages":"0670"},"PeriodicalIF":11.0,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12038163/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144021492","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Marine N-3 Fatty Acids Mitigate Hyperglycemia in Prediabetes by Improving Muscular Glucose Transporter 4 Translocation and Glucose Homeostasis.","authors":"Haoyu Li, Pan Zhuang, Xiaohui Liu, Yin Li, Yang Ao, Yimei Tian, Wei Jia, Yu Zhang, Jingjing Jiao","doi":"10.34133/research.0683","DOIUrl":"https://doi.org/10.34133/research.0683","url":null,"abstract":"<p><p>Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) have been proposed to benefit cardiometabolic health. However, the relationship between the intake of DHA and EPA and type 2 diabetes (T2D) risk remains equivocal, and the effects of DHA and EPA on skeletal muscle, the primary organ for glucose metabolism, merit further investigation. Here, we show that habitual fish oil supplementation was associated with a 9% lower T2D risk and significantly interacted with variants at GLUT4 in a prospective cohort of 48,358 people with prediabetes. Muscular metabolome analysis in the animal study revealed that DHA and EPA altered branched-chain amino acids, creatine, and glucose oxidation-related metabolites, concurrently with elevated muscular glycogen synthase and pyruvate dehydrogenase contents that promoted glucose disposal. Further myotube investigation revealed that DHA and EPA promoted muscular GLUT4 translocation by elevating Rab GTPases and target-SNARE expression. Together, DHA and EPA supplementation provides a promising approach for T2D prevention through targeting muscular glucose homeostasis, including enhancing GLUT4 translocation, glycogen synthesis, and aerobic glycolysis.</p>","PeriodicalId":21120,"journal":{"name":"Research","volume":"8 ","pages":"0683"},"PeriodicalIF":11.0,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12038161/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144009331","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Three-Dimensional Printable Color-Modulation and Shape-Programmable Structures: An Encryption Key for Image Recognition Electronic Locks.","authors":"Beibei Du, Xiayu Zhang, Teng Wang, Yunfei He, Mingyao Shen, Tao Yu","doi":"10.34133/research.0666","DOIUrl":"https://doi.org/10.34133/research.0666","url":null,"abstract":"<p><p>Stimuli-responsive materials have shown promising applications in the areas of sensing, bioimaging, information encryption, and bioinspired camouflage. In particular, multi-stimuli-responsive materials represent a hot topic due to their modulated properties under multiple stimuli. Herein, we successfully developed multi-stimuli-responsive inks and a series of complex multi-stimuli-responsive 3-dimensional (3D) structures were fabricated via digital light processing 3D-printing technology. Notably, these complex 3D structures show shape memory, fast-response photochromic and thermochromic behavior, and excellent repeatability due to the combination of photochromic molecules (4-(2,2-bis(4-fluorophenyl)vinyl) benzyl methacrylate) and thermochromic pigments. Furthermore, a programmable encrypted box that changes colors and morphology by controlling temperature and ultraviolet irradiation was designed and printed, and this encrypted box exhibits strong security using OpenCV-based image recognition technology. This strategy provides a promising approach for the design of multi-stimuli-responsive materials and complex encryption systems in the future.</p>","PeriodicalId":21120,"journal":{"name":"Research","volume":"8 ","pages":"0666"},"PeriodicalIF":11.0,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12022503/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143996386","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Low-Intensity Pulsed Ultrasound Promotes Oligodendrocyte Maturation and Remyelination by Down-regulating the Interleukin-17A/Notch1 Signaling Pathway in Mice with Ischemic Stroke.","authors":"Jingjing Wang, Yuxiao Gao, Bin Wang, Cong Zhang, Yi Yuan, Renhao Xu, Hui Ji, Xiangjian Zhang","doi":"10.34133/research.0676","DOIUrl":"https://doi.org/10.34133/research.0676","url":null,"abstract":"<p><p>Increasing evidence indicates that oligodendrocyte (OL) numbers and myelin as a dynamic cellular compartment perform a key role in the maintenance of neuronal function. Inhibiting white matter (WM) demyelination or promoting remyelination has garnered interest for its potential therapeutic strategy against ischemic stroke. Our previous work has shown that low-intensity pulsed ultrasound (LIPUS) could improve stroke recovery. However, it is unclear whether LIPUS can maintain WM integrity early after stroke or promote late WM repair. This study evaluated the efficacy of LIPUS on WM repair and long-term neurologic recovery after stroke. Male adult C57BL/6 mice underwent a focal cerebral ischemia model and were randomized to receive ultrasound stimulation (30 min once daily for 14 days). The effect of LIPUS on sensorimotor function was assessed by modified neurological severity score, rotarod test, grip strength test, and gait analysis up to 28 days after stroke. We found that ischemic stroke-induced WM damage was severe on day 7 and partially recovered on day 28. LIPUS prevented neuronal and oligodendrocyte progenitor cell (OPC) death during the acute phase of stroke (d7), protected WM integrity, and reduced brain atrophy and tissue damage during the recovery phase (d28). To further confirm the effect of LIPUS on remyelination, we assessed the proliferation and differentiation of OPCs. We found that LIPUS did not increase the number of OPCs (PDGFRα⁺ or NG2⁺), but markedly increased the number of newly produced mature OLs (APC⁺) and myelin protein levels. Mechanistically, LIPUS may promote OL maturation and remyelination by down-regulating the interleukin-17A/Notch1 signaling pathway. In summary, LIPUS can protect OLs and neurons early after stroke and promote long-term WM repair and functional recovery. LIPUS will be a viable strategy for the treatment of ischemic stroke in the future.</p>","PeriodicalId":21120,"journal":{"name":"Research","volume":"8 ","pages":"0676"},"PeriodicalIF":11.0,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12022504/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144042447","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Alanine Derived from <i>Ruminococcus_E bovis</i> Alleviates Energy Metabolic Disorders during the Peripartum Period by Providing Glucogenic Precursors.","authors":"Fanlin Kong, Shuo Wang, Yijia Zhang, Chen Li, Dongwen Dai, Yajing Wang, Zhijun Cao, Hongjian Yang, Shengli Li, Wei Wang","doi":"10.34133/research.0682","DOIUrl":"https://doi.org/10.34133/research.0682","url":null,"abstract":"<p><p>Peripartum dairy cows commonly experience energy metabolism disorders, which lead to passive culling of postpartum cows and a decrease in milk quality. By using ketosis peripartum dairy cows as a model, this study aims to elucidate the metabolic mechanism of peripartum cows and provide a novel way for managing energy metabolic disorders. From a cohort of 211 cows, we integrated multi-omics data (metagenomics, metabolomics, and transcriptomics) to identify key microbes and then utilized an in vitro rumen fermentation simulation system and ketogenic hepatic cells to validate the potential mechanisms and the effects of postbiotics derived from key microbes. Postpartum cows with metabolic disorders compensate for glucose deficiency through mobilizing muscle proteins, which leads to marked decreases in milk protein content. Concurrently, these cows experience rumen microbiota disturbance, with marked decreases in the concentrations of volatile fatty acids and microbial protein, and the deficiency of alanine (Ala) in microbial protein is correlated with the metabolic disorder phenotype. Metagenomic binning and in vitro fermentation assays reveal that <i>Ruminococcus_E bovis</i> (MAG 189) is enriched in amino acid biosynthesis functions and responsible for Ala synthesis. Furthermore, transcriptomic and metabolomic analyses of the liver in metabolic disorder cows also show impaired amino acid metabolism. Supplementation with Ala can alleviate ketogenesis in liver cell models by activating the gluconeogenesis pathway. This study reveals that <i>Ruminococcus_E bovis</i> is associated with host energy metabolism homeostasis by supplying glucogenic precursors to the liver and suggests the use of Ala as a method for the treatment of energy metabolism disorders in peripartum cows.</p>","PeriodicalId":21120,"journal":{"name":"Research","volume":"8 ","pages":"0682"},"PeriodicalIF":11.0,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12022398/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144043909","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multifunctional Magnetic Catheter Robot with Triaxial Force Sensing Capability for Minimally Invasive Surgery.","authors":"Shixiong Fu, Shiyuan Dong, Haolan Shen, Zhiqiang Chen, Guoyao Ma, Mingxue Cai, Chenyang Huang, Qianbi Peng, Chenyao Bai, Yuming Dong, Huanhuan Liu, Tianyu Yang, Tiantian Xu","doi":"10.34133/research.0681","DOIUrl":"https://doi.org/10.34133/research.0681","url":null,"abstract":"<p><p>Magnetic continuum robots offer flexibility and controllability, making them promising for minimally invasive surgery (MIS). However, the clinical application of these robots is relatively limited due to the difficulty of integrating miniaturized triaxial force sensors and their single functionality. This paper proposes a multifunctional magnetic catheter robot with magnetic actuation steering and triaxial force-sensing capabilities. The robot features 3 channels at its tip that integrate multi-segmented magnets, a novel triaxial force sensor, and various functional instruments. The sensor is calibrated, demonstrating high sensitivity and accuracy. The steering characterization of the robot confirms that the catheter tip exhibits effective flexibility and force sensing. Palpation experiments involving various hard lumps are performed on porcine kidney, with results verifying that the robot can reliably detect abnormal hard lumps within tissues. Additionally, palpation experiments in bronchial phantom demonstrate the robot's imaging and palpation capabilities for lung nodules with an integrated endoscope. Further, the robot, equipped with biopsy forceps, successfully performs palpation and biopsy functions on simulated stomach polyps, demonstrating its capability for effective tissue manipulation. By leveraging force-sensing capabilities and integrating multifunctional instruments, the robot shows potential for expanded applications in MIS, paving the way for important advancements in clinical procedures.</p>","PeriodicalId":21120,"journal":{"name":"Research","volume":"8 ","pages":"0681"},"PeriodicalIF":11.0,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12018763/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144049832","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic Homeostasis of Immune Cells Modulates Cardiovascular Diseases.","authors":"Mohan Li, Xiaolei Sun, Linqi Zeng, Aijun Sun, Junbo Ge","doi":"10.34133/research.0679","DOIUrl":"https://doi.org/10.34133/research.0679","url":null,"abstract":"<p><p>Recent investigations into the mechanisms underlying inflammation have highlighted the pivotal role of immune cells in regulating cardiac pathophysiology. Notably, these immune cells modulate cardiac processes through alternations in intracellular metabolism, including glycolysis and oxidative phosphorylation, whereas the extracellular metabolic environment is changed during cardiovascular disease, influencing function of immune cells. This dynamic interaction between immune cells and their metabolic environment has given rise to the novel concept of \"immune metabolism\". Consequently, both the extracellular and intracellular metabolic environment modulate the equilibrium between anti- and pro-inflammatory responses. This regulatory mechanism subsequently influences the processes of myocardial ischemia, cardiac fibrosis, and cardiac remodeling, ultimately leading to a series of cardiovascular events. This review examines how local microenvironmental and systemic environmental changes induce metabolic reprogramming in immune cells and explores the subsequent effects of aberrant activation or polarization of immune cells in the progression of cardiovascular disease. Finally, we discuss potential therapeutic strategies targeting metabolism to counteract abnormal immune activation.</p>","PeriodicalId":21120,"journal":{"name":"Research","volume":"8 ","pages":"0679"},"PeriodicalIF":11.0,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12015101/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144021855","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Noninvasive Self-Powered Iontophoresis Mask Based on a Water-Driven Fiber Battery.","authors":"Yiwen Wang, Yalin Tang, Ming Li, Tong Xu, Xuyan Lu, Deteng Zhang, Ning Yu, Mingwei Tian","doi":"10.34133/research.0667","DOIUrl":"https://doi.org/10.34133/research.0667","url":null,"abstract":"<p><p>Facial masks are often used to treat skin problems, and the introduction of microcurrent ion penetration technology can improve drug penetration and help facial tissue repair. However, most microcurrent stimulation masks contain a direct current power supply and require external power sources, resulting in inconvenient portability and use. Herein, we provide a noninvasive self-powered iontophoresis mask with a water-driven power supply, which is continuously prepared by self-constructing equipment to continually construct a zinc-manganese fiber battery (Zn-Mn@FB) and then seamlessly integrated with a nonwoven cellulose-based superabsorbent fiber substrate. The mask can be activated by water and is simple and portable to use. Zn-Mn@FB demonstrated a capacitance retention of 65.22% (1,000 cycles) and a specific discharge capacity of 27.33 mAh/g (10 cm), which improved with an increase in battery length to up to 41 mAh/g (30 cm). The iontophoresis mask exhibited a stable current within the safe range of 0.09 to 0.59 mA (within 800 s) after water activation, and the drug penetration area increased by 102.64%. The platform is expected to become a practical device for enhanced transdermal drug delivery in the medical field, with the potential to integrate additional components for expanded functionality and productization in the future.</p>","PeriodicalId":21120,"journal":{"name":"Research","volume":"8 ","pages":"0667"},"PeriodicalIF":11.0,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12015099/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144042448","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gecko Toe Pad-Inspired Robotic Gripper with Rapidly and Precisely Tunable Adhesion.","authors":"Shuai Li, Hongmiao Tian, Xijing Zhu, Mingxin Liu, Xiangmeng Li, Jinyou Shao","doi":"10.34133/research.0687","DOIUrl":"https://doi.org/10.34133/research.0687","url":null,"abstract":"<p><p>Gecko-inspired dry adhesives have shown great potential in the field of robotics. However, there is still a large gap between current artificial adhesive-based grippers and natural geckos, especially in terms of precise and fast control of adhesion, which is an important capability for robotic gripper systems, since the targets to be gripped may vary in size and weight (including thin, fragile, soft, and deformable), and manipulation must be fast to meet high productivity requirements. Here, we propose a robotic gripper that is able to switch adhesion rapidly (in less than 0.5 s) to grasp and release objects of various sizes and weights (such as glass substrates, fragile silicon wafers, and deformable polyethylene terephthalate films) by mimicking the self-peeling behavior of gecko toe pads. The gripper retains the fast and stable manipulation of the conventional mechanical gripper, which is more reliable and has a higher load capacity than stimulus-responsive switchable adhesives. Systematic experimental and theoretical studies provide insights into the construction and analysis of the self-peeling model and mechanism to identify certain crucial parameters affecting the self-peeling behavior. Furthermore, a strategy for active adhesion control (i.e., precise adhesion modulation) is integrated by introducing a preset peeling angle <i>θ_B</i> , providing the gripper with a quantitative criterion for adjusting the adhesion strength (0 to 82.77 kPa) according to the requirements of practical applications. The gripper has great potential to be an alternative end-operating gripper for robotic systems, opening an avenue for the development of robotic manipulation.</p>","PeriodicalId":21120,"journal":{"name":"Research","volume":"8 ","pages":"0687"},"PeriodicalIF":11.0,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12015100/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144014485","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}