Jove-Journal of Visualized Experiments最新文献_第9页

Massively Parallel Splicing Assay to Examine Splicing Errors Caused by Disease-Related Intronic Variants. 大规模平行剪接试验检查由疾病相关内含子变异引起的剪接错误。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-09-09 DOI: 10.3791/68984

Hung-Lun Chiang, Ang-Chu Huang, Shao-Wei Pan, Jia-Ying Su, Yen-Tsung Huang, Chien-Ling Lin

{"title":"Massively Parallel Splicing Assay to Examine Splicing Errors Caused by Disease-Related Intronic Variants.","authors":"Hung-Lun Chiang, Ang-Chu Huang, Shao-Wei Pan, Jia-Ying Su, Yen-Tsung Huang, Chien-Ling Lin","doi":"10.3791/68984","DOIUrl":"https://doi.org/10.3791/68984","url":null,"abstract":"Splicing errors represent 10-30% of the pathogenic mutations responsible for rare genetic disorders. RNA splicing ensures proper gene expression by selectively joining exons and removing introns, with key regulatory sequences being located within the introns. The 5' splice site and branch site interact with small nuclear RNAs to form the spliceosome's recognition complex, while elements such as the polypyrimidine tract and splicing enhancers/silencers recruit proteins to regulate spliceosome assembly. Predicting splicing disruptions from intronic variants is challenging due to the complexity of these interactions. Intronic variants, comprising 90% of natural human gene variations, may disrupt canonical splicing and give rise to disease. To investigate this possibility, we developed a massively parallel splicing assay (MaPSy) to assess patient-identified intronic variants. Synthesized oligonucleotides with reference or variant sequences were ligated into splicing minigenes containing promoter and polyadenylation signals. Each construct included two constant exons flanking a middle exon that harbored the variable intron-exon junction sequence of interest. The cellular splicing efficiency of the variant sequences was compared to reference counterparts, allowing us to identify significant disruptions as splicing variants. The results of the MaPSy can be validated through additional approaches, such as minigene assays or CRISPR-mediated genome editing in vivo. Furthermore, aggregate analysis of the disrupted junctions can provide deeper insights into splicing mechanisms and the molecular basis of diseases associated with splicing errors.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145193642","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Advancing Surgical Pathology Reporting in Head and Neck Cancer through 3D Visualization, Dynamic Documentation, and Optimized Communication. 通过3D可视化、动态文档和优化沟通推进头颈癌手术病理报告。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-09-09 DOI: 10.3791/68818

Sabrina R Comess, Justin K Joseph, Luke Stanisce, Margaret Brandwein-Weber, Michael Karasick, Mark L Urken

{"title":"Advancing Surgical Pathology Reporting in Head and Neck Cancer through 3D Visualization, Dynamic Documentation, and Optimized Communication.","authors":"Sabrina R Comess, Justin K Joseph, Luke Stanisce, Margaret Brandwein-Weber, Michael Karasick, Mark L Urken","doi":"10.3791/68818","DOIUrl":"https://doi.org/10.3791/68818","url":null,"abstract":"The successful surgical management of head and neck cancer relies on intraoperative communication between surgical and pathological teams to achieve oncologic clearance. Precise documentation of the actions taken and pathological information obtained during surgery is crucial for planning adjuvant treatment and to meaningfully interpret surveillance imaging. Achieving oncologic clearance of the cancer has significant implications with respect to reducing the risk of recurrence, reducing the need for multimodality adjuvant therapy, and improving quality of life. The surgical pathology report guides postoperative care, and yet it falls short of conveying the comprehensive body of information obtained at the time of surgery. We report the use of 3D scanning technology, principles of standardization, and software developments to improve the current surgical workflow and final pathology report. The approach described here integrates dynamic three-dimensional (3D) visuals, a series of intraoperative timeouts, annotated radiographs, and a novel surgical pathology reporting software to improve both intraoperative communication as well as postoperative understanding by medical and radiation oncologists. This unique methodology addresses shortcomings in the current and stagnant standard of care for pathologic documentation, paves the way for significant innovations in surgical pathology reporting, and holds the promise of improving patient outcomes.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145193652","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

White and Brown Adipose Grafts: An Approach to Correct Reproductive, Metabolic, and Renal Deficits in Black and Tan Brachyury (BTBR) Obese Mice. 白色和棕色脂肪移植：一种纠正黑色和棕色短腹（BTBR）肥胖小鼠生殖、代谢和肾脏缺陷的方法

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-09-09 DOI: 10.3791/68759

Melise Oliveira Mariano, Maria Theresa Araújo Balby-Rocha, Maria Clara Soares Klein, Bruno Serrano Barbosa, Marcella Liciani Franco, Andrei Furlan da Silva, Jaqueline de Carvalho Arjona, Gleice Regina Josefi-Rocha, Stephany Beyerstedt, Tuany Marques Reiter Alves, Érika Bevilaqua Rangel

{"title":"White and Brown Adipose Grafts: An Approach to Correct Reproductive, Metabolic, and Renal Deficits in Black and Tan Brachyury (BTBR) Obese Mice.","authors":"Melise Oliveira Mariano, Maria Theresa Araújo Balby-Rocha, Maria Clara Soares Klein, Bruno Serrano Barbosa, Marcella Liciani Franco, Andrei Furlan da Silva, Jaqueline de Carvalho Arjona, Gleice Regina Josefi-Rocha, Stephany Beyerstedt, Tuany Marques Reiter Alves, Érika Bevilaqua Rangel","doi":"10.3791/68759","DOIUrl":"https://doi.org/10.3791/68759","url":null,"abstract":"Transplantation of white adipose tissue (WAT) and brown adipose tissue (BAT) has emerged as a promising therapeutic strategy for reversing metabolic disorders without the need for sustained administration of exogenous agents. WAT functions as a key endocrine organ involved in energy homeostasis, while BAT is a thermogenic tissue critical for thermoregulation. The Black and Tan Brachyuric (BTBR) obese mouse model closely mimics key features of obesity, type 2 diabetes mellitus (T2DM), and diabetic kidney disease (DKD), including hyperglycemia, insulin resistance, hypometabolism, and subfertility. Using the protocol presented below, combined transplantation of WAT and BAT derived from BTBR wild-type and heterozygous donors into BTBR obese recipients resulted in the reversal of subfertility, attenuation of weight gain, normalization of glycemic control, and improvement of renal parameters. Notably, renal hypertrophy and apoptosis were reduced, while podocyte numbers were preserved. These findings suggest that combined WAT and BAT transplantation may represent an alternative therapeutic strategy for the treatment of T2DM, with concurrent benefits in mitigating DKD and obesity. Furthermore, this approach may aid in increasing the number of viable BTBR obese mice within vivarium colonies. Further investigation into the underlying mechanisms may facilitate the clinical translation of this intervention.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145193247","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

MTT-Based Normalization of Glycerol Release for Accurate Quantification of Lipolysis in Primary Rat Epididymal Adipocytes. 基于mtt的甘油释放归一化法准确定量大鼠附睾脂肪细胞脂肪溶解。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-09-09 DOI: 10.3791/68853

Leo Tsui, Peiying Ye, Shiling Xu, Bin Chen, Yuan Lin

{"title":"MTT-Based Normalization of Glycerol Release for Accurate Quantification of Lipolysis in Primary Rat Epididymal Adipocytes.","authors":"Leo Tsui, Peiying Ye, Shiling Xu, Bin Chen, Yuan Lin","doi":"10.3791/68853","DOIUrl":"https://doi.org/10.3791/68853","url":null,"abstract":"Lipolysis, a critical metabolic pathway, involves the hydrolysis of stored triglycerides into free fatty acids and glycerol. However, accurate quantification of glycerol output in lipid-rich primary adipocytes is often confounded by experimental artifacts, such as variability in cell viability and assay interference. To address these limitations, a straightforward method was established to normalize glycerol release measurements using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-based cell viability assay. This protocol leverages the difference in specific gravity between adipocytes and aqueous solutions, enabling simultaneous quantification of lipolytic output and cell viability. In drug screening experiments, pravastatin and simvastatin significantly enhanced glycerol release from abdominal adipocytes, whereas atorvastatin and lovastatin did not significantly alter glycerol output. These findings suggest that MTT-based normalization offers a robust and convenient approach for accurately quantifying lipolysis, with potential applications in drug discovery targeting obesity and metabolic syndrome. This study underscores the need to re-evaluate conventional lipolysis assays to distinguish genuine metabolic responses from nonspecific membrane leakage.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145193606","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Effect of Eye-Tracking Technology-Based Visual Scanning Training on Unilateral Spatial Neglect after Stroke. 基于眼动追踪技术的视觉扫描训练对脑卒中后单侧空间忽视的影响。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-09-09 DOI: 10.3791/68331

JinQin Zhang, XinPing Wang, FuBiao Huang, WenShuai Wang, YuQi Li, Min Zhang, FeiRu Chen, Bin Gu, Hao Yang, Yu Wei, WanNing Yang

{"title":"Effect of Eye-Tracking Technology-Based Visual Scanning Training on Unilateral Spatial Neglect after Stroke.","authors":"JinQin Zhang, XinPing Wang, FuBiao Huang, WenShuai Wang, YuQi Li, Min Zhang, FeiRu Chen, Bin Gu, Hao Yang, Yu Wei, WanNing Yang","doi":"10.3791/68331","DOIUrl":"https://doi.org/10.3791/68331","url":null,"abstract":"This study aimed to explore the effect of eye-tracking technology-based visual scanning training on recovery from unilateral spatial neglect after stroke. Stroke patients with unilateral spatial neglect (n = 48) from Beijing Bo'ai Hospital were recruited and randomly divided into an eye-tracking technology-based visual scanning training group (n = 24) and a conventional visual scanning training group (n = 24). The training regimen was 30 min/session, 1 session/day, and 5 days/week. The experimental group received visual scanning training via eye-tracking technology for 15 min and conventional unilateral spatial neglect training for 15 min. The control group received conventional unilateral spatial neglect training for 30 min. Both groups received conventional drug therapy and underwent conventional occupational rehabilitation. The Behaviour Inattention Test-Conventional Group (BIT-C), the Catherine Bergego Scale (CBS), and the Modified Barthel Index (MBI) were used to assess recovery from unilateral spatial neglect and to evaluate activities of daily living (ADLs) before and after treatment. The Mini-Mental State Examination (MMSE) was used to assess cognitive function before and after treatment. The results suggested that eye-tracking technology-based visual scanning training is more effective than conventional training in terms of alleviating unilateral spatial neglect and reducing the severity of neglect in ADLs. However, compared with conventional training, eye-tracking technology-based visual scanning training did not significantly increase ADL or MMSE scores.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145193663","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Renal Artery Function and Histopathology Linked to Plasma and Fecal Metabolites in Heart Failure Patients. 心力衰竭患者的肾动脉功能和组织病理学与血浆和粪便代谢物相关。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-09-09 DOI: 10.3791/68583

Junxia Zhang, Jingli Yang, Xuya Kang, Zeyuan Wang, Yusi Chen, Xinying Wang, Lin Yao, Yan Zhang, Yahan Liu, Erdan Dong

{"title":"Renal Artery Function and Histopathology Linked to Plasma and Fecal Metabolites in Heart Failure Patients.","authors":"Junxia Zhang, Jingli Yang, Xuya Kang, Zeyuan Wang, Yusi Chen, Xinying Wang, Lin Yao, Yan Zhang, Yahan Liu, Erdan Dong","doi":"10.3791/68583","DOIUrl":"https://doi.org/10.3791/68583","url":null,"abstract":"The gut microbiota and its associated host-microbe co-metabolites are increasingly recognized as key regulators of systemic metabolic balance and the cardiorenal axis, particularly within the context of cardiorenal syndrome. Although clinical evidence indicates that cardiac dysfunction may initiate or exacerbate renal pathological alterations, the key metabolic signaling molecules mediating the cardiorenal axis and their underlying mechanisms remain elusive. This study establishes a novel systematic research platform integrating cardiometabolic signatures with renal vascular function analysis, comprising: (1) Isolation and extraction of plasma/fecal metabolites from heart failure patients; (2) Ex vivo renal vascular isolation and primary culture techniques; (3) A multimodal evaluation framework for cardiorenal metabolic interactions, incorporating vascular functional assays, molecular biochemical tests for renal vascular injury markers, and histopathological analyses. Compared with healthy controls, metabolites from heart failure patients impaired renal vascular function and induced inflammatory responses, highlighting their potential as functional biomarkers in cardiorenal syndrome. This study does not focus on a specific metabolite; therefore, further identification and validation of the specific types of metabolites that exert pathogenic effects will be required in future studies using analytical techniques such as LC-MS/MS and NMR. Application of this platform revealed that cardiac disease-associated metabolites impair renal vascular function and homeostasis. These findings provide mechanistic insight into the metabolic drivers of cardiorenal interactions and offer a translational tool for identifying novel biomarkers and therapeutic targets.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145193704","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Personalized 3D-printed Headgear for Multi-electrode Transcranial Electrical Stimulation. 用于多电极经颅电刺激的个性化3d打印头套。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-09-09 DOI: 10.3791/68815

Benjamin M Hampstead, Annalise Rahman-Filipiak, Kayla Rinna, Stephen Schlaefflin, Kenneth Petscavage, Alexander R Guillen, Kamran Nazim, Jeffrey Moreno, Abhishek Datta, Marom Bikson, Alexandru D Iordan

{"title":"Personalized 3D-printed Headgear for Multi-electrode Transcranial Electrical Stimulation.","authors":"Benjamin M Hampstead, Annalise Rahman-Filipiak, Kayla Rinna, Stephen Schlaefflin, Kenneth Petscavage, Alexander R Guillen, Kamran Nazim, Jeffrey Moreno, Abhishek Datta, Marom Bikson, Alexandru D Iordan","doi":"10.3791/68815","DOIUrl":"https://doi.org/10.3791/68815","url":null,"abstract":"We present an overview of a novel, patented, approach for creating personalized 3D-printed headgear that enables multi-electrode transcranial electrical stimulation, even in the home. A structural magnetic resonance imaging (MRI) scan is used to optimize a montage that targets key brain areas/networks and is then uploaded to a secure server to design customized headgear that is ultimately 3D-printed. The headgear allows rapid multi-electrode placement (<1 min per electrode) with comparable accuracy as manual 10-10 measurements. We performed over 1,400 sessions across 63 older adults with neurological injury or disease. We describe an efficient training program for study partners (e.g., spouses) that enabled a 100% success rate and over 900 remotely administered sessions. Study partners and participants reported high levels of confidence and satisfaction with the experience. Because the electrode locations are based on personalized computational modeling, the headgear is ideal for techniques requiring precision (e.g., temporal interference stimulation, transcranial focused ultrasound, transcranial magnetic stimulation).","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145193746","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Erratum: A Modified Trier Social Stress Test for Vulnerable Mexican American Adolescents. 修正的特里尔社会压力测试对脆弱的墨西哥裔美国青少年。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-09-08 DOI: 10.3791/6640

Megan M Johnson, Julianna Deardorff, Kimberly Parra, Abbey Alkon, Brenda Eskenazi, Elizabeth Shirtcliff

引用次数: 0

Erratum: Optimizing and Evaluating Hand-drawing and Wet-spinning for Recombinant Spider Silk Fiber Production. 修正：优化和评价重组蜘蛛丝纤维生产的手绘和湿纺工艺。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-09-08 DOI: 10.3791/6649

Anupama Ghimire, Hina Batool, Sara Evans, Suad Rashid, Donovan L Rainey, Andrew A Y Chen, Xiang-Qin Liu, Jan K Rainey

引用次数: 0

Multi-Stage Vibration Control Mechanism and Experimental Effectiveness of Viscous Damper with Overload Protector. 带过载保护器的粘性阻尼器多级振动控制机理及实验效果。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2025-09-05 DOI: 10.3791/68410

Shiming Zhang, Yuan Wu, Kai Yang, Jiashuo Wang

{"title":"Multi-Stage Vibration Control Mechanism and Experimental Effectiveness of Viscous Damper with Overload Protector.","authors":"Shiming Zhang, Yuan Wu, Kai Yang, Jiashuo Wang","doi":"10.3791/68410","DOIUrl":"https://doi.org/10.3791/68410","url":null,"abstract":"This research presents an innovative hybrid seismic isolation system that integrates a viscous damper with overload protection (VD-OP) to improve seismic performance and overcome the limitations of traditional isolation methods. The VD-OP incorporates a displacement-dependent nonlinear damping mechanism and a force-limiting feature, effectively controlling excessive damping forces during high-velocity movements caused by varying seismic intensities. A mechanical model, calibrated using experimental data, is developed to replicate the nonlinear damping and overload protection characteristics of the VD-OP. Numerical simulations are validated through experimental testing, providing a solid foundation for optimizing parameters and evaluating performance. The findings show that the force-limiting function plays a critical role in reducing excessive forces on the isolation layer, improving isolation efficiency, and controlling acceleration responses in the superstructure. Parametric studies and case analyses across different seismic scenarios confirm the system's strong energy dissipation capabilities and adaptability, ensuring effective isolation during frequent earthquakes, controlled deformation under design-level events, and structural protection during severe earthquakes. The performance-based design approach proposed here offers practical guidance for the implementation of the VD-OP system, presenting a reliable solution to enhance the seismic resilience of engineering structures.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145126247","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0