Methods in molecular biology最新文献_第10页

Small Cell Lung Cancer: Screening of Kinase Inhibitors. 小细胞肺癌：激酶抑制剂的筛选。

Methods in molecular biology Pub Date : 2026-01-01 DOI: 10.1007/978-1-0716-5050-9_14

Hamilton Gerhard, Marie-Therese Eggerstorfer, Sandra Stickler

{"title":"Small Cell Lung Cancer: Screening of Kinase Inhibitors.","authors":"Hamilton Gerhard, Marie-Therese Eggerstorfer, Sandra Stickler","doi":"10.1007/978-1-0716-5050-9_14","DOIUrl":"10.1007/978-1-0716-5050-9_14","url":null,"abstract":"Small cell lung cancer (SCLC) exhibits extreme numbers of circulating tumor cells (CTCs) that are operative in tumor dissemination and possibly drug resistance, ultimately resulting in a dismal prognosis. Tumor cells depend on kinases, which are triggered by a range of growth factors for their malignant proliferation. Insulin-like growth factor-I receptor (IGF-1) and vascular endothelial growth factor receptor (VEGFR), c-Kit, epidermal growth factor receptor (EGFR), as well as downstream signaling transmitters such as phosphoinositide 3-kinase (PI3K), Akt and the mammalian target of rapamycin (mTOR) and a range of other kinases were investigated as potential anti-tumor targets in SCLC. Despite the expression of rational targets, the studies with tyrosine kinase inhibitors (TKIs) in SCLC have been very disappointing in the clinic. The preclinical activity of kinase inhibitors against native and resistant SCLC cell lines failed to predict efficacy in patients. Screening of a kinase inhibitor library showed activity of most compounds against single SCLC CTC cells but high chemoresistance of BHGc10 spheroids (termed tumorospheres). Only a few hydrophobic multitarget inhibitors proved active against the spheroids, and drugs that failed in clinical trials targeting IGF-1R, AURORA kinase and Polo kinases showed low activity against CTC tumorospheres. These results demonstrate that for SCLC, kinase inhibitors should be tested against spheroids to obtain a better prediction of their potential clinical activity.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2999 ","pages":"171-181"},"PeriodicalIF":0.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148422451","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

Allosteric Network Analysis Toolkit for Single-Domain Phosphoproteins. 单域磷酸化蛋白变构网络分析工具包。

Methods in molecular biology Pub Date : 2026-01-01 DOI: 10.1007/978-1-0716-4828-5_9

Maham Hamid, Safee Ullah Chaudhary, Alessandro Pandini, Shahid M Khan

引用次数: 0

The Genus Boechera as a Model for Apomixis Research. 作为无融合研究的一个模式的布氏属。

Methods in molecular biology Pub Date : 2026-01-01 DOI: 10.1007/978-1-0716-5001-1_15

Terezie Mandáková

{"title":"The Genus Boechera as a Model for Apomixis Research.","authors":"Terezie Mandáková","doi":"10.1007/978-1-0716-5001-1_15","DOIUrl":"10.1007/978-1-0716-5001-1_15","url":null,"abstract":"The genus Boechera, a prominent member of the tribe Boechereae within the family Brassicaceae, has emerged as an exceptional model for apomixis research, owing to its unique evolutionary, reproductive, and genomic characteristics. With over 480 genetically distinct taxa and a distribution spanning North America, Greenland, and parts of Asia, Boechera exemplifies remarkable ecological and genetic diversity. A hallmark of the genus is its high frequency of gametophytic apomixis, including diplospory and apospory, which, combined with hybridization, drive genetic innovation and environmental adaptability. Moreover, Boechera is notable for its diploid expression of apomixis, independent of polyploidy, providing an unparalleled system for dissecting asexual reproduction. Genomic adaptations, such as heterochromatic supernumerary chromosomes, further highlight its evolutionary complexity. Recent advances, including the discovery of critical regulatory genes like APOLLO and UPGRADE2, have deepened our understanding of the genetic basis of apomixis. These insights position Boechera as a cornerstone for elucidating apomictic pathways and leveraging these mechanisms for crop improvement. The genus not only enhances our understanding of apomixis but also offers transformative potential for agriculture.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2987 ","pages":"221-229"},"PeriodicalIF":0.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145409475","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

Analyzing Endolysosome-Mitochondria Membrane Contact Sites by Thin Section TEM of Human Fibroblasts. 人成纤维细胞内溶酶体-线粒体膜接触部位的电镜分析。

Methods in molecular biology Pub Date : 2026-01-01 DOI: 10.1007/978-1-0716-4844-5_10

Claudia Parra-Ruiz, Carlos Enrich, Silvana Zanlungo

引用次数: 0

Modeling Lysosomal Disease in Dictyostelium discoideum: Examining the Trafficking and Secretion of Lysosomal Enzymes. 盘状盘基网线菌溶酶体疾病的建模：检测溶酶体酶的运输和分泌。

Methods in molecular biology Pub Date : 2026-01-01 DOI: 10.1007/978-1-0716-4844-5_14

William D Kim, Robert J Huber

引用次数: 0

Label-Free Imaging of Lysosomes in Living Cells by Photothermal Microscopy. 光热显微镜下活细胞中溶酶体的无标记成像。

Methods in molecular biology Pub Date : 2026-01-01 DOI: 10.1007/978-1-0716-4844-5_4

Jun Miyazaki, Ryotaro Wagatsuma

引用次数: 0

Lysosome Enrichment Using Superparamagnetic Iron Oxide Nanoparticles (SPIONs). 利用超顺磁性氧化铁纳米颗粒富集溶酶体。

Methods in molecular biology Pub Date : 2026-01-01 DOI: 10.1007/978-1-0716-4844-5_7

Pathma Muthukottiappan, Sara Bonini, Dominic Winter

引用次数: 0

Adapting Komagataella phaffii Cultivation Media to REACH Regulations. 法菲小松菌培养基适应REACH法规。

Methods in molecular biology Pub Date : 2026-01-01 DOI: 10.1007/978-1-0716-4779-0_30

Julian Kopp, Alexander Pekarsky, Oliver Spadiut

引用次数: 0

Auxin-Induced Degradation of Target Proteins in Pichia pastoris. 毕赤酵母中生长素诱导的靶蛋白降解。

Methods in molecular biology Pub Date : 2026-01-01 DOI: 10.1007/978-1-0716-4779-0_6

Leonie Lehmayer, Lukas Bernauer, Anita Emmerstorfer-Augustin

{"title":"Auxin-Induced Degradation of Target Proteins in Pichia pastoris.","authors":"Leonie Lehmayer, Lukas Bernauer, Anita Emmerstorfer-Augustin","doi":"10.1007/978-1-0716-4779-0_6","DOIUrl":"10.1007/978-1-0716-4779-0_6","url":null,"abstract":"The disruption of protein production is a major approach used for investigating protein function in a cell. In yeasts, this is mostly achieved by introducing an insertion or deletion (\"indel\") into a certain gene or by disrupting the whole reading frame. However, these methods have noteworthy limitations. For example, whole gene deletions may influence promoter or terminator regions of genes located up- and downstream of this particular gene; essential genes cannot simply be knocked out in order to study their function; and the overall absence of a protein can allow time for compensatory mechanisms that confound interpretation of results. To overcome these limitations, this chapter presents the application of the auxin-inducible degron (AID) technology for fast and efficient degradation of a target protein in Komagataella phaffii. The AID system is based on the formation of a chimeric SCF complex with the plant F-box protein TIR1, which, in the presence of auxin, leads to the polyubiquitination of the target sequence, the so-called AID tag. If the AID tag is fused to a protein of interest, this protein is immediately subjected to ubiquitin-mediated proteasome degradation upon incubation with auxin. In this work, the usability and efficiency of auxin-inducible degradation of Gut1, a well-studied glycerol kinase with an important role in glycerol metabolism, is demonstrated. Important parameters for experimental design for the use of this system in K. phaffii are being evaluated. The results prove that this toolkit can be expanded to all areas of K. phaffii-related research.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2697 ","pages":"101-118"},"PeriodicalIF":0.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145199985","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

Direct Reprogramming of Microglia into Neurons. 小胶质细胞直接重编程成神经元。

Methods in molecular biology Pub Date : 2026-01-01 DOI: 10.1007/978-1-0716-4807-0_13

Kanae Matsuda-Ito, Eriko Hatai, Takashi Irie, Taito Matsuda, Kinichi Nakashima

{"title":"Direct Reprogramming of Microglia into Neurons.","authors":"Kanae Matsuda-Ito, Eriko Hatai, Takashi Irie, Taito Matsuda, Kinichi Nakashima","doi":"10.1007/978-1-0716-4807-0_13","DOIUrl":"10.1007/978-1-0716-4807-0_13","url":null,"abstract":"In recent years, direct reprogramming methods have attracted attention as they can bypass the immature stem cell state and directly induce the desired cell type by introducing key transcription factors into somatic cells. This technique offers advantages such as shorter induction times compared to inducing the desired cells from induced pluripotent stem cells. Furthermore, this approach enables the provision of the requisite cell types to patients requiring treatment without eliciting an immune rejection response. Microglia, the resident immune cells of the central nervous system, exhibit high proliferative and regenerative capacities, allowing them to repopulate even after almost complete depletion. In response to injury, microglia accumulate in damaged brain regions and become the primary cell type that forms the glial scar. Given these properties, microglia are promising candidates for direct reprogramming into neurons in the injured area without depleting cell sources. This chapter outlines a protocol for directly reprogramming cortical microglia into neurons, utilizing primary culture and lentiviral-mediated gene transfer techniques.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2974 ","pages":"153-161"},"PeriodicalIF":0.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145582451","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