Methods in molecular biology最新文献_第7页

Use of an Intramolecular Quenched Fluorescence (IQF) Cleavage Assay for Assessing Enzyme Kinetics of Gamma-Secretase in Human Skin Fibroblasts and Keratinocytes. 使用分子内淬灭荧光（IQF）切割试验评估人皮肤成纤维细胞和角质形成细胞中γ -分泌酶的酶动力学。

Methods in molecular biology Pub Date : 2025-01-09 DOI: 10.1007/7651_2024_587

Beita Badiei, Luis A Garza

引用次数: 0

Ring Magnet-Guided Magnetic Manipulation for Biofabrication of 3D Cellular Structures. 环形磁体引导的三维细胞结构生物制造的磁操作。

Methods in molecular biology Pub Date : 2025-01-09 DOI: 10.1007/7651_2024_597

Muge Anil-Inevi, Engin Ozcivici

引用次数: 0

3D Cell Culture Models as a Platform for Studying Tumor Progression, Testing Treatment Responses, and Discovering Biomarkers. 3D细胞培养模型作为研究肿瘤进展，测试治疗反应和发现生物标志物的平台。

Methods in molecular biology Pub Date : 2025-01-09 DOI: 10.1007/7651_2024_595

Peyda Korhan, Ezgi Bağırsakçı, Yasemin Öztemur Islakoğlu, Neşe Atabey

{"title":"3D Cell Culture Models as a Platform for Studying Tumor Progression, Testing Treatment Responses, and Discovering Biomarkers.","authors":"Peyda Korhan, Ezgi Bağırsakçı, Yasemin Öztemur Islakoğlu, Neşe Atabey","doi":"10.1007/7651_2024_595","DOIUrl":"https://doi.org/10.1007/7651_2024_595","url":null,"abstract":"In this chapter, we present a detailed protocol for establishing a three-dimensional (3D) multicellular tumor spheroids (MCTSs) model to simulate the tumor microenvironment (ME) associated with metabolic dysfunction-associated steatotic liver disease (MASLD) for the study of hepatocellular carcinoma (HCC) and colorectal cancer (CRC) cell aggressiveness, growth, and metastasis potential. The MASLD microenvironment (MASLD-ME) is recreated by embedding hepatic stellate cells in a collagen I matrix within a Boyden chamber system. The metabolic medium mimics MASLD conditions, enriched with high glucose, fructose, insulin, and fatty acids, to simulate metabolic stresses associated with the disease.In the protocol, cancer cells are loaded in the upper compartment to analyze their migration toward the MASLD-ME, thereby facilitating studies on cancer cell invasiveness and metastatic capacity. This method offers an adaptable, reproducible model to research disease progression and investigate therapeutic interventions, contributing to preclinical research on MASLD-related liver cancer pathophysiology and potential drug responses.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142951265","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Enzymatically Ligated Nucleic Acid Nanocapsules for the Delivery of Therapeutic Nucleic Acids and Small Molecule Drugs. 用于输送治疗性核酸和小分子药物的酶联核酸纳米胶囊。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4402-7_4

Jenna N Cannata, Jessica L Rouge

引用次数: 0

Precision Cell-Cell Assembly Through Light-Mediated DNA Interactions. 精确细胞-细胞组装通过光介导的DNA相互作用。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4402-7_11

Katelyn Mathis, Brian Meckes

引用次数: 0

Surface Functionalization of Elastomers with Biopolymers. 弹性体与生物聚合物的表面功能化。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4402-7_13

Emilie Morin, Elana Muzzy, Andrea S Carlini

引用次数: 0

Nonhematopoietic Stem Cell Identification and Sorting in Adult Retina. 成人视网膜非造血干细胞的鉴定与分选。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4386-0_5

Ana Flores, Laura Fernández-Sánchez, Oksana Kutsyr, Alberto Yáñez, María Luisa Gil, Daniel Gozalbo, Victoria Maneu, Pedro Lax

引用次数: 0

Subcellular Localization of Geminivirus Proteins by Laser Scanning Confocal Microscopy. 用激光扫描共聚焦显微镜研究双病毒蛋白的亚细胞定位。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4454-6_18

Christiane Eliza Motta Duarte, João Paulo Batista Machado, Bianca Gouveia-Mageste, Fredy Davi Albuquerque Silva, Elizabeth Pacheco Batista Fontes

{"title":"Subcellular Localization of Geminivirus Proteins by Laser Scanning Confocal Microscopy.","authors":"Christiane Eliza Motta Duarte, João Paulo Batista Machado, Bianca Gouveia-Mageste, Fredy Davi Albuquerque Silva, Elizabeth Pacheco Batista Fontes","doi":"10.1007/978-1-0716-4454-6_18","DOIUrl":"10.1007/978-1-0716-4454-6_18","url":null,"abstract":"In eukaryotic cells, the subcellular localization of proteins is inherently linked to their function. Since viruses rely on the host cellular machinery to complete their life cycle, viral proteins are expected to employ the host transport machinery to reach various compartments. Several factors, including the multifunctional nature of viral proteins, the stage of virus infection, and interactions with both viral and host proteins, influence the final destination of viral proteins. For instance, NSP (nuclear shuttle protein) from bipartite begomoviruses and CP (coat protein) from monopartite begomoviruses typically exhibit nuclear localization, yet their subcellular distribution can vary depending on coexpression partners and stage of infection. Virtually all viral proteins display dynamic subcellular distribution patterns that change under their specific functions at different stages of the virus life cycle. Thus, identifying the subcellular distribution of viral proteins is essential for comprehending their multiple roles during infection. This chapter outlines a protocol for efficiently determining the subcellular localization of viral proteins during infection or when expressed with protein partners. The protocol essentially consists of three steps: (i) cloning the viral protein and protein partners fused to fluorescent tags, (ii) transiently expressing the tagged proteins in N. benthamiana leaves, and (iii) determining the subcellular localization of the tagged proteins using confocal microscopy.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2912 ","pages":"205-226"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143597373","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Assessment of Bacterial Contamination in Semen. 精液中细菌污染的评估

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4406-5_39

Ferran Garriga, Sergi Bonet, Marc Yeste

{"title":"Assessment of Bacterial Contamination in Semen.","authors":"Ferran Garriga, Sergi Bonet, Marc Yeste","doi":"10.1007/978-1-0716-4406-5_39","DOIUrl":"10.1007/978-1-0716-4406-5_39","url":null,"abstract":"Semen contamination is one of the main issues of concern in livestock industry. While the presence of some bacteria in semen is considered as normal, high bacterial loads detrimentally affect sperm quality and fertilizing ability, thus having a negative repercussion on the efficiency of artificial insemination. For this reason, the present chapter focuses on the methods used for the assessment of semen contamination in farm animals. Routinely, bacterial culture is performed to assess the overall degree of contamination in sperm samples; in some cases, however, the identification of bacterial species is needed to determine the optimal antibiotic composition of semen preservation media and address whether those bacteria have developed antibiotic resistance. In addition, other techniques, such as scanning electron microscopy, can be used to evaluate sperm-bacteria interaction. This chapter also discusses the strategies to prevent bacteria contamination in semen. As the ejaculate is not a sterile fluid, media for preservation usually include, as aforementioned, antibiotics to avoid bacterial growth. Because of the mounting restriction in the usage of antibiotics, which is due to the increase of bacterial resistance, other approaches to prevent bacterial contamination of semen include preservation at low temperatures (5 °C) and the inclusion of alternative molecules such as antimicrobial peptides.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2897 ","pages":"591-600"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143811695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Live-Cell Single-Molecule Imaging of Influenza A Virus-Receptor Interaction. 甲型流感病毒与受体相互作用的活细胞单分子成像。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4326-6_4

Lukas Broich, Yang Fu, Christian Sieben

引用次数: 0