Frontiers in Bioengineering and Biotechnology最新文献

{"title":"sEMG-based gesture recognition using multi-stream adaptive CNNs with integrated residual modules.","authors":"Yutong Xia, Dawei Qiu, Cheng Zhang, Jing Liu","doi":"10.3389/fbioe.2025.1487020","DOIUrl":"10.3389/fbioe.2025.1487020","url":null,"abstract":"<p><strong>Introduction: </strong>Convolutional neural networks are widely used in gesture recognition research, which employs surface electromyography. However, when processing surface electromyography data, current deep learning models still face challenges, such as insufficient effective feature extraction, poor performance in multi-gesture recognition, and low accuracy in recognizing sparse surface electromyography.</p><p><strong>Methods: </strong>To address these issues, this study proposed a multi-stream adaptive convolutional neural networks with residual modules (MSACNN-RM) for surface electromyography gesture recognition, which integrates multiple streams of convolutional neural networks, adaptive convolutional neural networks, and residual modules to enhance the model's feature extraction and learning capabilities. This improves the model's ability to extract and understand complex data patterns.</p><p><strong>Results: </strong>The experimental results demonstrated that the model achieved recognition accuracies of 98.24%, 93.52%, and 92.27% respectively on the Ninapro DB1, Ninapro DB2, and Ninapro DB4 datasets. Compared with other deep learning models, MSACNN-RM achieves higher accuracy compared to existing models.</p><p><strong>Discussion: </strong>The proposed model explores features of sparse sEMG signals by leveraging multi-stream convolution, the combination of adaptive convolution modules and ResNet blocks enhances the model's ability of extracting crucial gesture features. In the future, in order to deal with differences in sEMG signals caused by variations among individuals, a universal multi-gesture recognition algorithm should be developed. Meanwhile, the model should focus on optimizing and streamlining the network to reduce computational load.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1487020"},"PeriodicalIF":4.3,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12069338/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143975854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Wireless skin sensors for electrocardiogram and heart rate monitoring in the neonatal intensive care unit: a prospective feasibility, safety, and accuracy study.","authors":"Eva Senechal, Daniel Radeschi, Shasha Lv, Emilie Jeanne, Ana Saveedra Ruiz, Lydia Tao, Brittany Dulmage, Wissam Shalish, Robert Edward Kearney, Guilherme Sant'Anna","doi":"10.3389/fbioe.2025.1555882","DOIUrl":"10.3389/fbioe.2025.1555882","url":null,"abstract":"<p><strong>Objectives: </strong>Assess feasibility, safety, and accuracy of electrocardiogram (ECG) and heart rate (HR) monitoring in neonates, using a new wireless skin sensor.</p><p><strong>Methods: </strong>Prospective observational study in infants of any gestational age admitted in the neonatal intensive care unit. ECG/HR signals were simultaneously recorded from a standard wired and new wireless system with bedside annotations. Feasibility was evaluated as signal coverage, gap numbers/durations, and sources of gaps. Safety was appraised by changes in skin condition and pain after/upon wireless sensor removal. Accuracy was measured using bias and 95% limits of agreement, and the coefficient of determination. The ability of the wireless sensors to detect normal and abnormal HR values was evaluated using a Clark Error Grid. Additionally, user satisfaction from parents and nurses were appraised using a short questionnaire.</p><p><strong>Results: </strong>25 infants had 757 h of recorded signals over 96 days. ECG coverage was 99.9% [IQR: 99.9%-99.95%] for the wired vs 97.8% [IQR: 81.6%-99.9%; p < 0.00] for the wireless system, while HR coverage was 99.4% [IQR: 98.6%-99.9%] vs 89.7% [IQR: 75.6%-97.6%; p < 0.00]. Wireless ECG gaps were <5 s in 97% of cases, and HR gaps <30 s in 85%. All ECG gaps and 57% of HR gaps were due to Bluetooth disconnection (BD). 78% of BD in wireless HR were during kangaroo care (78%). Of 192 skin photographs (96 pairs), 98% were taken, showing increased but low skin scores post-removal, with median pain scores also low. Accuracy metrics showed strong agreement, with the Clark Error Grid indicating 97% of paired signals led to the same clinical outcome. Among 23 nurse and 18 parent responses, satisfaction with the wireless system was high.</p><p><strong>Conclusion: </strong>ECG and HR monitoring using a new wireless skin sensor was feasible, safe, and accurate when compared to the wired standard. Future adjustments in the technology are needed to improve signal coverage during handling and KC and test the sensors in unstable and more immature patients. Limitations included challenges in recruiting unstable neonates, variability introduced by multiple raters completing pain assessments, and inability to apply safety metrics to the wired standard of care.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1555882"},"PeriodicalIF":4.3,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12069355/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143989023","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Scalability of spheroid-derived small extracellular vesicles production in stirred systems.","authors":"Thibaud Dauphin, Laurence de Beaurepaire, Apolline Salama, Quentin Pruvost, Clémentine Claire, Karine Haurogné, Sophie Sourice, Aurélien Dupont, Jean-Marie Bach, Julie Hervé, Eric Olmos, Steffi Bosch, Blandine Lieubeau, Mathilde Mosser","doi":"10.3389/fbioe.2025.1516482","DOIUrl":"10.3389/fbioe.2025.1516482","url":null,"abstract":"<p><strong>Introduction: </strong>Small extracellular vesicle (sEV)-based therapies have gained widespread interest, but challenges persist to ensure standardization and high-scale production. Implementing upstream processes in a chemically defined media in stirred-tank bioreactors (STBr) is mandatory to closely control the cell environment, and to scale-up production, but it remains a significant challenge for anchorage-dependent cells.</p><p><strong>Methods: </strong>We used a human β cell line, grown as monolayer or in suspension as spheroid in stirred systems. We assessed the consequences of culturing these cells in 3D with, or without fetal bovine serum in a chemically defined medium, for cell growth, viability and metabolism. We next explored how different scale-up strategies might influence cell and spheroid formation in spinner flask, with the aim to transfer the process in instrumented Ambr®250 STBr. Lastly, we analyzed and characterized sEV production in monolayer, spinner flask and STBr.</p><p><strong>Results and discussion: </strong>Generation of spheroids in a chemically defined medium allowed the culture of highly viable cells in suspension in stirred systems. Spheroid size depended on the system's volumetric power input (P/V), and maintaining this parameter constant during scale-up proved to be the optimal strategy for standardizing the process. However, transferring the spinner flask (SpF) process to the Ambr®250 STBr at constant P/V modified spheroid size, due to important geometric differences and impeller design. Compared to a monolayer reference process, sEV yield decreased two-fold in SpF, but increased two-fold in STBr. Additionally, a lower expression of the CD63 tetraspanin was observed in sEV produced in both stirred systems, suggesting a reduced release of exosomes compared to ectosomes. This study addresses the main issues encountered in spheroid culture scale-up in stirred systems, rather conducive for the production of ectosomes.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1516482"},"PeriodicalIF":4.3,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12069995/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143996765","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selective laser melting fabrication of functionally graded macro-porous Ti-6Al-4V scaffold for cavity bone defect reconstruction.","authors":"Zhuangzhuang Li, Yi Luo, Ruicheng Liu, Shanfang Zou, Yitian Wang, Taojun Gong, Xuanhong He, Yong Zhou, Minxun Lu, Li Min, Chongqi Tu","doi":"10.3389/fbioe.2025.1550309","DOIUrl":"https://doi.org/10.3389/fbioe.2025.1550309","url":null,"abstract":"<p><p>Reconstruction of cavitary bone defects poses significant challenges in orthopedic surgery due to the irregular shapes and compromised mechanical properties of surrounding bone. This study developed a functionally graded macro-porous scaffold (FGMPS) using selective laser melting (SLM) for cavitary bone defect reconstruction. The FGMPS featured a porosity gradient (74%-86%) and macropores ≥1,600 µm, mimicking the natural density gradient of cancellous bone. Micro-CT analysis confirmed high structural fidelity and interconnected porosity. Compression tests in two orientations revealed distinct stress-strain responses: vertically aligned gradients (FGMPS-V) exhibited sequential layer engagement, while horizontally aligned gradients (FGMPS-H) demonstrated higher stiffness and strength due to uniform load distribution. The elastic modulus ranged from 383 MPa (FGMPS-V) to 577 MPa (FGMPS-H), with yield strength of 22-40 MPa, aligning well with cancellous bone properties. These findings highlight the FGMPS's potential to offer a promising solution for cavitary bone defect repair.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1550309"},"PeriodicalIF":4.3,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12066661/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143961657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mechanical property changes of glial LC and RGC axons in response to high intraocular pressure.","authors":"Bochao Ma, Liu Liu, Yushu Liu, Jifeng Ren, Xiuqing Qian","doi":"10.3389/fbioe.2025.1574231","DOIUrl":"https://doi.org/10.3389/fbioe.2025.1574231","url":null,"abstract":"<p><strong>Introduction: </strong>Pathological high intraocular pressure (IOP) is an important risk factor for glaucoma. The lamina cribrosa (LC) area in the optic nerve head is the initial site of optic nerve injury for glaucoma. LC deformation caused by elevated IOP will compress the retinal ganglion cells (RGC) axons passing through it, thereby leading to the damage of the RGC axons. The deformation of LC is highly correlated with its mechanical properties. Therefore, changes in mechanical properties of LC with the duration of high IOP is of great significance.</p><p><strong>Methods: </strong>To investigate the impact of chronic high IOP on the mechanical properties of the LC, rat models were established by cauterizing the superior scleral vein and injecting 5-fluorouracil (5-FU) under the conjunctiva to maintain elevated IOP. The linear elastic properties of the glial LC and RGC axons in affected eyes were measured using atomic force microscopy (AFM) combined with image segmentation techniques. Morphological alterations of the glial LC were assessed using hematoxylin-eosin staining, immunofluorescence staining, and transmission electron microscopy (TEM).</p><p><strong>Results: </strong>Compared to the control group, the Young's modulus of the glial LC decreased by 35.5%, 74.2%, and 80.6% at 4, 8, and 12 weeks of elevated IOP, respectively. Similarly, the Young's modulus of RGC axons decreased by 45.6%, 70.9%, and 75.9% over the same time points. These findings demonstrate a time-dependent reduction in the mechanical stiffness of both glial LC and RGC axons under chronic high IOP conditions.</p><p><strong>Discussion: </strong>The progressive decrease in Young's modulus indicated that prolonged high IOP compromises the structural integrity and mechanical properties of the LC and RGC axons. This mechanical weakening likely contributes to the pathophysiological process of optic nerve injury in glaucoma. The present study offers important insights into the biomechanical mechanisms underlying glaucomatous damage, which may guide future research and therapeutic strategies.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1574231"},"PeriodicalIF":4.3,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12066477/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144009149","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Urine-derived stem cells: a sustainable resource for advancing personalized medicine and dental regeneration.","authors":"Gamal A Atia, Ahmed Abdal Dayem, Ehab S Taher, Wafaa Y Alghonemy, Ssang-Goo Cho, Ahmed A Aldarmahi, Md Azizul Haque, Abeer Alshambky, Noha Taymour, Ateya M Ibrahim, Donia E Zaghamir, Ekramy M Elmorsy, Helal F Hetta, Mohamed E Mohamed, Kasim S Abass, Shifan Khanday, Ahmed Abdeen","doi":"10.3389/fbioe.2025.1571066","DOIUrl":"https://doi.org/10.3389/fbioe.2025.1571066","url":null,"abstract":"<p><p>Urine-based therapy, an ancient practice, has been utilized across numerous civilizations to address a wide range of ailments. Urine was considered a priceless resource in numerous traditional therapeutic applications due to its reported medicinal capabilities. While the utilization of urine treatment is contentious and lacks significant support from modern healthcare, the discovery of urine-derived stem cells (UDSCs) has introduced a promising avenue for cell-based therapy. UDSCs offer a noninvasive and easily repeatable collection method, making them a practical and viable option for therapeutic applications. Research has shown that UDSCs contribute to organ preservation by promoting revascularization and decreasing inflammatory reactions in many diseases and conditions. This review will outline the contemporary status of UDSCs research and explore their potential applications in both fundamental science and medical practice.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1571066"},"PeriodicalIF":4.3,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12066649/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144009193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Structural and functional analysis of a homotrimeric collagen peptide.","authors":"Xinling Zhang, Kexin Li, Nan Lu, Takafumi Takebayashi, Boyu Zhou, Hongbin Xie, Yufan Li, Xingyun Long, Xingjiong Qin, Hongyi Zhao, Jiying Dong","doi":"10.3389/fbioe.2025.1575341","DOIUrl":"https://doi.org/10.3389/fbioe.2025.1575341","url":null,"abstract":"<p><strong>Objective: </strong>This study aimed to chemically synthesize a homotrimeric collagen peptide, evaluate its safety, and assess its effectiveness in promoting collagen synthesis.</p><p><strong>Methods: </strong>A homotrimeric collagen peptide was synthesized and structurally characterized using circular dichroism and infrared spectroscopy. Thermal stability was analyzed by TG-DSC, and molecular weight and amino acid composition were determined. <i>In vitro</i> cytotoxicity testing assessed safety, while UV-induced photoaging experiments evaluated its effects on collagen and elastin synthesis. <i>In vivo</i> studies in BALB/c mice examined its impact on collagen content, skin structure, and angiogenesis.</p><p><strong>Results: </strong>The synthesized collagen peptide exhibited high purity (99.1%) and an amino acid composition of glycine, proline, and hydroxyproline in a balanced ratio (15:17:13). Structural analysis confirmed a stable triple-helical conformation similar to type I collagen with excellent thermal stability (Tm = 326.15°C). Cytotoxicity testing showed no adverse effects on cell viability. <i>In vitro</i>, the peptide significantly enhanced collagen and elastin synthesis in fibroblasts. <i>In vivo</i>, intradermal and subcutaneous injection increased collagen content, improved skin structure, and enhanced microvessel density.</p><p><strong>Conclusion: </strong>This study presents a chemically synthesized homotrimeric collagen peptide with superior purity, structural stability, and biological efficacy in promoting collagen synthesis. Compared to previous studies, this biomimetic material exhibits exceptional thermal stability (Tm = 326.15°C) and a well-balanced amino acid composition, enabling applications in cosmetics and medical devices requiring heat sterilization (e.g., autoclaving), as validated by our patented method (China Patent No. ZL202410309842.9).</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1575341"},"PeriodicalIF":4.3,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12066645/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144006668","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Imrecoxib attenuates osteoarthritis by modulating synovial macrophage polarization through inactivating COX-2/PGE2 signaling pathway.","authors":"Peng Peng, Wanling Zheng, Yuchen Liu, Jingyuan Huang, Bin Zhang, Jiawei Shen, Jiangang Cao","doi":"10.3389/fbioe.2025.1526092","DOIUrl":"https://doi.org/10.3389/fbioe.2025.1526092","url":null,"abstract":"<p><strong>Introduction: </strong>Although biomaterials strategies have been regarded as a promising approach for the treatment of osteoarthritis (OA), identifying novel drugs to be delivered for modulate macrophage polarization is still unclear. As a commonly used non-steroidal anti-inflammatory drug for OA, Imrecoxib may be a novel drug to direct and sustain macrophage phenotype. However, the specific protective mechanism of Imrecoxib in OA remains unclear. This study aims to investigate whether Imrecoxib would treat OA by regulating synovial macrophage polarization.</p><p><strong>Methods: </strong>The research involves constructing mouse destabilization of medial meniscus (DMM) model to assess the changes in pain, bone destruction, cartilage degeneration, and synovial macrophage phenotypes following Imrecoxib treatment. Additionally, the effects of macrophage conditioned medium (CM) pretreated with Imrecoxib on the chondrocyte apoptosis, inflammation and degeneration-related factor expression were evaluated. The role of COX-2/PGE2 signaling pathway in the macrophage phenotype changes was further investigated.</p><p><strong>Results: </strong>We found that Imrecoxib alleviated pain, cartilage degeneration and synovitis, promoted polarization of M1 macrophages toward M2 phenotype <i>in vivo</i> and <i>in vitro</i>. <i>In vitro</i> experiments, Imrecoxib-CM protected chondrocyte by modulating macrophage polarization. Furthermore, Imrecoxib regulates macrophage polarization through the COX-2/PGE2 pathway.</p><p><strong>Conclusion: </strong>This study unravels that Imrecoxib protects joint cartilage and attenuates osteoarthritis by modulating synovial macrophage polarization through inactivating COX-2/PGE2 signaling pathway, providing new drug delivery strategy for the clinical treatment of OA.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1526092"},"PeriodicalIF":4.3,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12066670/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143994459","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Innovative modification strategies and emerging applications of natural hydrogel scaffolds for osteoporotic bone defect regeneration.","authors":"Yanan Chen, Qinghua Zhao","doi":"10.3389/fbioe.2025.1591896","DOIUrl":"https://doi.org/10.3389/fbioe.2025.1591896","url":null,"abstract":"<p><p>Osteoporosis, a prevalent systemic metabolic bone disease, is characterized by diminished bone mass, microarchitectural deterioration of bone tissue, and heightened bone fragility. In osteoporotic patients, chronic and progressive bone loss often leads to fractures and, in advanced cases, critical-sized bone defects. While traditional bone repair approaches are constrained by significant limitations, the advent of bioactive scaffolds has transformed the therapeutic paradigm for osteoporotic bone regeneration. Among these innovations, natural polymer-based hydrogel scaffolds have emerged as a particularly promising solution in bone tissue engineering, owing to their superior biocompatibility, tunable biodegradation properties, and exceptional ability to replicate the native extracellular matrix environment. This review systematically explores recent breakthroughs in modification techniques and therapeutic applications of natural hydrogel scaffolds for osteoporotic bone defect repair, while critically analyzing existing clinical challenges and proposing future research trajectories in this rapidly evolving field.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1591896"},"PeriodicalIF":4.3,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12066444/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144007617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A versatile genetic toolkit for engineering <i>Wickerhamomyces ciferrii</i> for tetraacetyl phytosphingosine production.","authors":"Seong-Rae Lee, Jun Su Kang, Pyung Cheon Lee","doi":"10.3389/fbioe.2025.1586218","DOIUrl":"https://doi.org/10.3389/fbioe.2025.1586218","url":null,"abstract":"<p><p><i>Wickerhamomyces ciferrii</i>: a non-conventional yeast with significant industrial potential for tetraacetyl phytosphingosine (TAPS), remains underutilized due to the lack of a comprehensive genetic toolbox. In this study, we developed a modular genetic system tailored for <i>Wickerhamomyces ciferrii</i> to enable strain engineering and metabolic pathway optimization. This toolkit includes episomal plasmids incorporating multiple selectable markers, replication origins, and fluorescent reporters. Systematic evaluation of four antibiotic resistance markers demonstrated that nourseothricin, geneticin, and zeocin effectively confer resistance, whereas hygromycin B did not support selection in this host. Among three tested replication origins, 2μ and CEN6/ARS4 enabled stable episomal maintenance, whereas panARS failed to replicate. Expression analysis of six fluorescent proteins under the endogenous <i>PGK1</i> promoter revealed significant variability in transcript levels, which correlated with codon adaptation index values, emphasizing the importance of codon optimization for heterologous expression. Additionally, characterization of the endogenous <i>TDH3, PGK1,</i> and <i>PDA1</i> promoters using two highly expressed fluorescent proteins confirmed that promoter strength is largely independent of the downstream coding sequence. To demonstrate the functional application of this toolkit, we overexpressed a phosphorylation-insensitive mutant of acetyl-CoA carboxylase (<i>ACC1</i> ^{<i>S26A-S1161A</i>} ), resulting in a 2.4-fold increase in TAPS production. Collectively, this study establishes a versatile genetic platform for <i>W. ciferrii</i>, providing a robust foundation for future synthetic biology and metabolic engineering applications.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1586218"},"PeriodicalIF":4.3,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12066694/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143991456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}