Methods in molecular biology最新文献_第6页

Principles of Indirect Co-culture Method Using Transwell. 使用 Transwell 间接共培养法的原理

Methods in molecular biology Pub Date : 2024-03-20 DOI: 10.1007/7651_2024_537

Mohammad Rasouli, Fatemeh Safari

引用次数: 0

Evaluation of Autophagy in Conjunctival Fibroblasts. 评估结膜成纤维细胞的自噬作用

Methods in molecular biology Pub Date : 2024-03-19 DOI: 10.1007/7651_2024_523

Parvaneh Mehrbod, Paola Brun, Umberto Rosani, Andrea Leonardi, Saeid Ghavami

引用次数: 0

Measurement of Autophagic Activity in Cancer Cells with Flow Cytometric Analysis Using Cyto-ID Staining. 利用 Cyto-ID 染色流式细胞分析法测量癌细胞的自噬活性

Methods in molecular biology Pub Date : 2024-03-07 DOI: 10.1007/7651_2024_526

Merve Şansaçar, Emel Başak Gencer Akçok

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A Lipidomics Approach to Determine the Role of Lipids and Its Crosstalk with Autophagy in Lung Cancer Metastasis. 用脂质组学方法确定脂质在肺癌转移中的作用及其与自噬的相互关系

Methods in molecular biology Pub Date : 2024-03-06 DOI: 10.1007/7651_2024_524

Simone C da Silva Rosa, Javad Alizadeh, Rui Vitorino, Arun Surendran, Amir Ravandi, Biniam Kidane, Saeid Ghavami

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Involvement of Autophagic Machinery in Neuropathogenesis: Targeting and Relevant Methods of Detection. 自噬机制参与神经发病：靶向和相关检测方法。

Methods in molecular biology Pub Date : 2024-03-06 DOI: 10.1007/7651_2024_516

Nourhan Sayed, Alaa Emam Ali, Doaa Mokhtar Elsherbiny, Samar S Azab

{"title":"Involvement of Autophagic Machinery in Neuropathogenesis: Targeting and Relevant Methods of Detection.","authors":"Nourhan Sayed, Alaa Emam Ali, Doaa Mokhtar Elsherbiny, Samar S Azab","doi":"10.1007/7651_2024_516","DOIUrl":"https://doi.org/10.1007/7651_2024_516","url":null,"abstract":"The exquisite balance between cellular prosurvival and death pathways is extremely necessary for homeostasis. Different forms of programmed cell death have been widely studied and reported such as apoptosis, necroptosis, pyroptosis, and autophagy. Autophagy is a catabolic process important for normal cellular functioning. The main aim of this machinery is to degrade the misfolded or damaged proteins, unuseful organelles, and pathogens, which invade the cells, thereby maintaining cellular homeostasis and assuring the regular renewal of cell components. This prosurvival function of autophagy highlights its importance in many human diseases, as the disturbance of this tightly organized process ultimately causes detrimental effects. Interestingly, neurons are particularly susceptible to damage upon the presence of any alteration in the basal level of the autophagic activity; this could be due to their high metabolic demand, post-mitotic nature, and the contribution of autophagy in the different fundamental functions of neurons. Herein, we have reported the role of autophagy in different CNS disorders such as Parkinson's disease, Alzheimer's disease, Huntington's disease, and epilepsy, besides the pharmacological agents targeting autophagy. Due to the significant contribution of autophagy in the pathogenesis of many diseases, it is crucial to develop effective methods to detect this dynamic process. In this chapter, we have summarized the most frequently employed techniques in studying and detecting autophagy including electron microscopy, fluorescence microscopy, Western blotting, intracellular protein degradation, and sequestration assay.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140028358","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

Investigating Protein-Protein Interactions of Autophagy-Involved TNIP1. 调查自噬参与的 TNIP1 蛋白质与蛋白质之间的相互作用

Methods in molecular biology Pub Date : 2024-03-06 DOI: 10.1007/7651_2024_525

Michael L Samulevich, Liam E Carman, Brian J Aneskievich

引用次数: 0

Assessment of Autophagy in Leishmania Parasites. 评估利什曼寄生虫的自噬作用

Methods in molecular biology Pub Date : 2024-03-06 DOI: 10.1007/7651_2024_517

Somtochukwu S Onwah, Jude E Uzonna, Saeid Ghavami

{"title":"Assessment of Autophagy in Leishmania Parasites.","authors":"Somtochukwu S Onwah, Jude E Uzonna, Saeid Ghavami","doi":"10.1007/7651_2024_517","DOIUrl":"https://doi.org/10.1007/7651_2024_517","url":null,"abstract":"Leishmaniasis is a neglected tropical disease caused by numerous species of Leishmania parasites, including Leishmania major. The parasite is transmitted by several species of sandfly vectors and infects myeloid cells leading to a myriad of inflammatory responses, immune dysregulations, and disease manifestations. Every cell undergoes autophagy, a self-regulated degradative process that permits the cells to recycle damaged or worn-out organelles in order to maintain cellular health and homeostasis. Studies have shown that Leishmania modulates their host cell autophagic machinery and there are indications that the parasite-specific autophagic processes may be valuable for parasite virulence and survival. However, the role of autophagy in Leishmania is inconclusive because of the limited tools available to study the Leishmania-specific autophagic machinery. Here, we describe methods to study and definitively confirm autophagy in Leishmania major. Transmission electron microscopy (TEM) allowed us to visualize Leishmania autophagosomes, especially those containing damaged mitochondrial content, as well as dividing mitochondria with ongoing fusion/fission processes. Flow cytometry enabled us to identify the amount of acridine orange dye accumulating in the acidic vacuolar compartments in Leishmania major by detecting fluorescence in the red laser when autophagic inhibitors or enhancers were included. These methods will advance studies that aim to understand autophagic regulation in Leishmania parasites that could provide insights into developing improved therapeutic targets against leishmaniasis.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140028356","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

Principles of Hanging Drop Method (Spheroid Formation) in Cell Culture. 细胞培养中的悬滴法（球体形成）原理。

Methods in molecular biology Pub Date : 2024-02-28 DOI: 10.1007/7651_2024_527

Mohammad Rasouli, Fatemeh Safari, Mohammad Hossein Kanani, Hiva Ahvati

{"title":"Principles of Hanging Drop Method (Spheroid Formation) in Cell Culture.","authors":"Mohammad Rasouli, Fatemeh Safari, Mohammad Hossein Kanani, Hiva Ahvati","doi":"10.1007/7651_2024_527","DOIUrl":"https://doi.org/10.1007/7651_2024_527","url":null,"abstract":"A type of three-dimensional (3D) cell culture models which is simple and easy is hanging drop method. The hanging drop method emerges as a pivotal technique with diverse applications in cancer research and cell biology. This method facilitates the formation of multicellular spheroids, providing a unique environment for studying cell behavior dynamics. The hanging drop method's theoretical underpinning relies on gravity-enforced self-assembly, allowing for cost-effective, reproducible 3D cell cultures with controlled spheroid sizes. The advantages of this approach include its efficiency in producing cellular heterogeneity, particularly in non-adherent 3D cultures, and its ability to create hypoxic spheroids, making it a suitable model for studying cancer. Moreover, the hanging drop method has proven valuable in investigating various aspects such as tissue structure, signaling pathways, immune activation of cancer cells, and notably, cell proliferation. Researchers have utilized the hanging drop method to explore the dynamics of cell proliferation, studying the effects of mesenchymal stem cells (MSC) secretome on cancer cells. The method's application involves co-culturing different cell lines, assessing spheroid formations, and quantifying their sizes over time. These studies have unveiled intricate cell behavior dynamics, demonstrating how the MSC secretome influences cancer cell growth and viability within a three-dimensional co-culture paradigm.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139972612","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

Visualizing Lipophagy as a New Mechanism of the Synthesis of Sex Steroids in Human Ovary and Testis Using Immunofluorescence Staining Method. 用免疫荧光染色法观察嗜脂性是人类卵巢和睾丸合成性激素的新机制

Methods in molecular biology Pub Date : 2024-02-28 DOI: 10.1007/7651_2024_520

Yashar Esmaeilian, Sevgi Yusufoglu, Ece Iltumur, Gamze Bildik, Ozgur Oktem

引用次数: 0

Real-Time Visualization of Cholesterol Trafficking in Human Granulosa Cells Using Confocal Live Cell Microscopy as a Tool to Study the Novel Role of Autophagy in Sex Steroid Synthesis. 利用共聚焦活细胞显微镜实时观察人类颗粒细胞中胆固醇的迁移，研究自噬在性类固醇合成中的新作用。

Methods in molecular biology Pub Date : 2024-02-28 DOI: 10.1007/7651_2024_521

Yashar Esmaeilian, Ozgur Oktem

引用次数: 0