Methods in cell biology最新文献_第6页

Using C. elegans as a model for neurodegenerative diseases: Methodology and evaluation. 将优雅蛇作为神经退行性疾病的模型：方法与评估。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-04-25 DOI: 10.1016/bs.mcb.2024.03.004

Elena Caldero-Escudero, Silvia Romero-Sanz, Sergio De la Fuente

引用次数: 0

Assessment of translation rate in leukemic cells and immune cells of the microenvironment by OPP protein synthesis assay. 通过 OPP 蛋白合成试验评估白血病细胞和微环境免疫细胞的翻译率。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-07-02 DOI: 10.1016/bs.mcb.2024.06.006

Vanessa Klapp, Ozgu Gumustekin, Jerome Paggetti, Etienne Moussay, Anne Largeot

{"title":"Assessment of translation rate in leukemic cells and immune cells of the microenvironment by OPP protein synthesis assay.","authors":"Vanessa Klapp, Ozgu Gumustekin, Jerome Paggetti, Etienne Moussay, Anne Largeot","doi":"10.1016/bs.mcb.2024.06.006","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.06.006","url":null,"abstract":"Despite being tightly regulated, messenger RNA (mRNA) translation, a manner in which cells control expression of genes and rapidly respond to stimuli, is highly dysfunctional and plastic in pathologies including cancer. Conversely, the investigation of molecular mechanisms whereby mRNA translation becomes aberrant in cancer, as well as inhibition thereof, become critical in developing novel therapeutic approaches. More specifically, in malignancies such as chronic lymphocytic leukemia in which aberrant global and transcript specific translation has been linked with poorer patient outcomes, targeting translation is a relevant approach, with various translation inhibitors under development. Here we elaborate on a protein synthesis assay by flow cytometry, O-propargyl-puromycin, demonstrating global mRNA translation rate with a variety of different applications including cell lines, primary cells or co-culture systems in vitro. This method provides a comprehensive tool in quantifying the rate of global mRNA translation in cancer cells, as well as that of the tumor microenvironment cells, or in response to inhibitory therapeutic agents while offering the possibility to simultaneously assess other cellular markers.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142406523","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Enzyme-linked ImmunoSpot (ELISpot) assay to quantify peptide-specific IFN-γ production by splenocytes in a mouse tumor model after radiation therapy. 用酶联免疫斑点（ELISpot）测定法量化放疗后小鼠肿瘤模型脾细胞产生的肽特异性 IFN-γ。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-09-05 DOI: 10.1016/bs.mcb.2024.07.001

Benoît Lecoester, Yaoyao Xie, Amélie Marguier, Laura Boulerot, Marine Malfroy, Olivier Adotévi, Jihane Boustani

引用次数: 0

Animal models of neglected parasitic diseases: In vivo multimodal imaging of experimental trypanosomatid infections. 被忽视寄生虫病的动物模型：实验性锥虫感染的体内多模态成像。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-05-24 DOI: 10.1016/bs.mcb.2024.04.003

Jean Marc Ngoune Tsagmo, Brice Rotureau, Estefanía Calvo Alvarez

{"title":"Animal models of neglected parasitic diseases: In vivo multimodal imaging of experimental trypanosomatid infections.","authors":"Jean Marc Ngoune Tsagmo, Brice Rotureau, Estefanía Calvo Alvarez","doi":"10.1016/bs.mcb.2024.04.003","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.04.003","url":null,"abstract":"African trypanosomiases and leishmaniases are significant neglected tropical diseases (NTDs) that affect millions globally, with severe health and socio-economic consequences, especially in endemic regions. Understanding the pathogenesis and dissemination of Trypanosoma brucei and Leishmania spp. parasites within their hosts is pivotal for the development of effective interventions. Whole-body bioluminescence and fluorescence imaging systems (BLI and FLI, respectively), are powerful tools to visualize and quantify the progression and distribution of these parasites in real-time within live animal models. By combining this technology with the engineering of stable T. brucei and Leishmania spp. strains expressing luciferase and/or fluorescent proteins, crucial aspects of the infection process including the parasites' homing, the infection dynamics, the tissue tropism, or the efficacy of experimental treatments and vaccines can be deeply investigated. This methodology allows for enhanced sensitivity and resolution, elucidating previously unrecognized infection niches and dynamics. Importantly, whole-body in vivo imaging is non-invasive, enabling for longitudinal studies during the course of an infection in the same animal, thereby aligning with the \"3Rs\" principle of animal research. Here, we detail a protocol for the generation of dual-reporter T. brucei and L. major, and their use to infect mice and follow the spatiotemporal dynamics of infection by in vivo imaging systems. Additionally, 3D micro-computed tomography (μCT) coupled to BLI in T. brucei-infected animals is applied to gain insights into the anatomical parasite distribution. This Chapter underscores the potential of these bioimaging modalities as indispensable tools in parasitology, paving the way for novel therapeutic strategies and deeper insights into host-parasite interactions.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141331341","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Spinal nerve ligation: An experimental model to study neuropathic pain in rats and mice. 脊神经结扎：研究大鼠和小鼠神经性疼痛的实验模型

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-04-25 DOI: 10.1016/bs.mcb.2024.03.006

Erick J Rodríguez-Palma, Itzel I Ramos-Rodríguez, Saúl Huerta de la Cruz, Vinicio Granados-Soto, Maria Sancho

{"title":"Spinal nerve ligation: An experimental model to study neuropathic pain in rats and mice.","authors":"Erick J Rodríguez-Palma, Itzel I Ramos-Rodríguez, Saúl Huerta de la Cruz, Vinicio Granados-Soto, Maria Sancho","doi":"10.1016/bs.mcb.2024.03.006","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.03.006","url":null,"abstract":"Neuropathic pain, defined as the most terrible of all tortures, which a nerve wound may inflict, is a common chronic painful condition caused by gradual damage or dysfunction of the somatosensory nervous system. As with many chronic diseases, neuropathic pain has a profound economic and emotional impact worldwide and represents a major public health issue from a treatment standpoint. This condition involves multiple sensory symptoms including impaired transmission and perception of noxious stimuli, burning, shooting, spontaneous pain, mechanical or thermal allodynia and hyperalgesia. Current pharmacological options for the treatment of neuropathic pain are limited, ineffective and have unacceptable side effects. In this framework, a deeper understanding of the pathophysiology and molecular mechanisms associated with neuropathic pain is key to the development of promising new therapeutical approaches. For this purpose, a plethora of experimental models that mimic common clinical features of human neuropathic pain have been characterized in rodents, with the spinal nerve ligation (SNL) model being one of the most widely used. In this chapter, we provide a detailed surgical procedure of the SNL model used to induce neuropathic pain in rats and mice. We further describe the behavioral approaches used for stimulus-evoked and spontaneous pain assessment in rodents. Finally, we demonstrate that our SNL model induces multiple pain behaviors in rats and mice.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141331348","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Flow cytometric detection of CD11b⁺ Gr-1⁺ cells in nontumor-bearing mice: A propolis-elicited model. 非肿瘤小鼠 CD11b+ Gr-1+ 细胞的流式细胞术检测：蜂胶诱导模型

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-10-11 DOI: 10.1016/bs.mcb.2023.05.010

Hiroshi Kitamura

引用次数: 0

Image processing and supervised machine learning for retinal microglia characterization in senescence. 用于衰老期视网膜小胶质细胞特征描述的图像处理和监督机器学习。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-01-28 DOI: 10.1016/bs.mcb.2022.12.008

Soyoung Choi, Daniel Hill, Jonathan Young, Maria Francesca Cordeiro

{"title":"Image processing and supervised machine learning for retinal microglia characterization in senescence.","authors":"Soyoung Choi, Daniel Hill, Jonathan Young, Maria Francesca Cordeiro","doi":"10.1016/bs.mcb.2022.12.008","DOIUrl":"10.1016/bs.mcb.2022.12.008","url":null,"abstract":"The process of senescence impairs the function of cells and can ultimately be a key factor in the development of disease. With an aging population, senescence-related diseases are increasing in prevalence. Therefore, understanding the mechanisms of cellular senescence within the central nervous system (CNS), including the retina, may yield new therapeutic pathways to slow or even prevent the development of neuro- and retinal degenerative diseases. One method of probing the changing functions of senescent retinal cells is to observe retinal microglial cells. Their morphological structure may change in response to their surrounding cellular environment. In this chapter, we show how microglial cells in the retina, which are implicated in aging and diseases of the CNS, can be identified, quantified, and classified into five distinct morphotypes using image processing and supervised machine learning algorithms. The process involves dissecting, staining, and mounting mouse retinas, before image capture via fluorescence microscopy. The resulting images can then be classified by morphotype using a support vector machine (SVM) we have recently described showing high accuracy. This SVM model uses shape metrics found to correspond with qualitative descriptions of the shape of each morphotype taken from existing literature. We encourage more objective and widespread use of methods of quantification such as this. We believe automatic delineation of the population of microglial cells in the retina, could potentially lead to their use as retinal imaging biomarkers for disease prediction in the future.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139672174","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Quantitative analysis of nuclear deformations and DNA damage foci dynamics by live-cell imaging. 通过活细胞成像对核变形和 DNA 损伤灶动态进行定量分析。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-01-09 DOI: 10.1016/bs.mcb.2022.12.010

Elena Faustini, Andrea Panza, Matteo Longaretti, Francisca Lottersberger

{"title":"Quantitative analysis of nuclear deformations and DNA damage foci dynamics by live-cell imaging.","authors":"Elena Faustini, Andrea Panza, Matteo Longaretti, Francisca Lottersberger","doi":"10.1016/bs.mcb.2022.12.010","DOIUrl":"10.1016/bs.mcb.2022.12.010","url":null,"abstract":"The correct repair of DNA Double Strand Breaks (DSBs) is fundamental to prevent the loss of genetic information, mutations, and chromosome rearrangements. An emerging determinant of DNA repair is chromatin mobility. However, how chromatin mobility can influence DSBs repair is still poorly understood. While increased mobility is generally associated with the correct repair by Homologous Recombination (HR) of DSBs generated in heterochromatin, it promotes the mis-repair of multiple distal DSBs by Non-Homologous End Joining (NHEJ). Here we describe a method for detecting and quantifying DSBs mobility by live-cell imaging in the context of multiple DSBs prone to mis-repair by NHEJ. In addition, we discuss a set of parameters that can be used for quantitative and qualitative analysis of nuclear deformations and to discard nuclei where the deformation could affect the analysis of DSBs mobility. While this method is based on the visualization of DSBs with the mCherry-53BP1-2 fusion protein, we believe that it can also be used to analyze the mobility of nuclear foci formed by different fluorescent proteins.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139741412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Visualizing DNA damage and repair using single molecule super resolution microscopy. 利用单分子超分辨率显微镜观察 DNA 损伤和修复。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-08-21 DOI: 10.1016/bs.mcb.2023.02.004

Sophie T B Morgan, Donna R Whelan, Ashley M Rozario

引用次数: 0

In vitro osteoclastogenesis assessment using murine myeloid-derived suppressor cells. 利用小鼠髓源性抑制细胞进行体外破骨细胞生成评估。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-10-14 DOI: 10.1016/bs.mcb.2023.04.007

Kyu Hwan Kwack, Lixia Zhang, Keith L Kirkwood

引用次数: 0