Methods in molecular biology最新文献_第10页

Dissecting the Heterogeneity of Senescent Cells Through CITE-seq Integration. 通过CITE-seq整合分析衰老细胞的异质性。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4434-8_7

Kotb Abdelmohsen, Krystyna Mazan-Mamczarz, Dimitrios Tsitsipatis, Martina Rossi, Rachel B Munk, Myriam Gorospe

{"title":"Dissecting the Heterogeneity of Senescent Cells Through CITE-seq Integration.","authors":"Kotb Abdelmohsen, Krystyna Mazan-Mamczarz, Dimitrios Tsitsipatis, Martina Rossi, Rachel B Munk, Myriam Gorospe","doi":"10.1007/978-1-0716-4434-8_7","DOIUrl":"https://doi.org/10.1007/978-1-0716-4434-8_7","url":null,"abstract":"Cellular senescence, a state of persistent growth arrest following cell damage, is associated with aging and age-related diseases. Understanding cell heterogeneity within senescent populations is crucial for developing therapies to mitigate senescence-associated pathologies. The protocol described here outlines an integrated approach to exploit the presence of cell surface proteins on subsets of senescent cells to study their heterogeneity at the single-cell level. After identifying senescence-associated surface proteins by mass spectrometry (MS) and then performing cellular indexing of transcriptomes and epitopes sequencing (CITE-seq) single-cell analysis, we were able to identify unique transcriptomic programs associated with specific surface protein markers expressed in some senescent cells but not in others. We illustrate the utility of this approach by investigating the complex heterogeneity of senescent cell populations. However, this methodology can be applied to other biological scenarios where cells with unique transcriptomic profiles can be studied individually, thanks to the presence of specific cell surface proteins that distinguish them from other cells within the same population.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2908 ","pages":"99-109"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143990074","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

Nanoparticles-Assisted Peptide Cancer Vaccine Delivery. 纳米颗粒辅助多肽癌症疫苗递送。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4542-0_10

Lanqing Luo, Rui Kuai

引用次数: 0

Micro-Focus Reduction Neutralization Tests (mFRNT) for Neutralizing Antibody (Nab) Quantification Against Yellow Fever Virus. 测定黄热病病毒中和抗体（Nab）的微焦点减少中和试验（mFRNT）

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4458-4_17

Adriana de Souza Azevedo, Nathalia Dos Santos Alves

引用次数: 0

A Method for H2A.Z/H2B Dimer Exchange Within Nucleosomes In Vitro. A H2A的方法。核小体内Z/H2B二聚体的体外交换。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4486-7_3

Benoit Guillemette, Liette Laflamme, Luc Gaudreau

引用次数: 0

Standardized Outcome Measures for the Clinical Application of Tissue Engineered Products. 组织工程产品临床应用的标准化结果测量。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4510-9_26

Anna S van den Bosch, Melinda Farkas, Frederique M Kemme, Matthea M Stoop, Paul P M van Zuijlen, Annebeth Meij-de Vries, Esther Middelkoop, Anouk Pijpe

{"title":"Standardized Outcome Measures for the Clinical Application of Tissue Engineered Products.","authors":"Anna S van den Bosch, Melinda Farkas, Frederique M Kemme, Matthea M Stoop, Paul P M van Zuijlen, Annebeth Meij-de Vries, Esther Middelkoop, Anouk Pijpe","doi":"10.1007/978-1-0716-4510-9_26","DOIUrl":"https://doi.org/10.1007/978-1-0716-4510-9_26","url":null,"abstract":"The clinical application of tissue engineered products aims to regenerate skin and enhance the appearance, texture, sensation, and functionality of affected skin or scars. To evaluate the efficacy and effectiveness of novel products, level 1 randomized controlled trials are required. However, currently a significant challenge in this field is the clinical and statistical heterogeneity of trial designs and outcome measures for tissue engineered products. To address this challenge and improve clinical outcomes, it is essential to standardize high-quality outcome measures. These measures are clinimetrically developed tools designed to reliably and validly assess specific health outcomes across diverse patient populations, clinical settings, and studies. This chapter proposes a core set of outcome measurements designed to evaluate the most relevant, feasible, reliable, and valid parameters in the clinical application of tissue engineered products. For routine clinical practice, we recommend the inclusion of photography, graft take, re-epithelialization, and scar quality assessment using the Patient and Observer Scar Assessment Scale. For clinical trials, additional measures such as colorimetry and elasticity assessment are advised. Detailed protocols for these methods are provided, resulting in a standardized core outcome set. High-quality outcome assessment also necessitates specialized training and the presence of trained personal at all assessment points. The chapter further addresses specific considerations for outcome assessment in pediatric patients, remote assessment strategies, and additional recommendations for optimizing clinical and research practices in this field. The implementation of standardized outcome measurement is essential for improving the outcomes of patients treated with tissue engineered products.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2922 ","pages":"335-354"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144001497","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

Flow Cytometry as Immunoassay Tool for Research on Yellow Fever Virus. 流式细胞术作为研究黄热病病毒的免疫分析工具。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4458-4_1

Raquel Curtinhas de Lima, Nieli Rodrigues da Costa Faria, Amanda Torrentes de Carvalho

{"title":"Flow Cytometry as Immunoassay Tool for Research on Yellow Fever Virus.","authors":"Raquel Curtinhas de Lima, Nieli Rodrigues da Costa Faria, Amanda Torrentes de Carvalho","doi":"10.1007/978-1-0716-4458-4_1","DOIUrl":"https://doi.org/10.1007/978-1-0716-4458-4_1","url":null,"abstract":"Flow cytometry is a sensitive and practical technique that can be applied in both basic and clinical research. It allows extracting quantitative and multiparametric valuable information to assist in the study of cell immunophenotyping, morphological complexity, location and/or expression of extra and intracellular molecules involved in metabolic and proliferative pathways, inflammation, viability, and cell death, among others. The parameters are analyzed by labeling antigens of yellow fever virus and/or cells with fluorescent specific monoclonal antibodies or dyes for immunophenotyping and data acquisition using a flow cytometer. In translational research, flow cytometry is a useful tool in tracking and monitoring the etiology, evolution, and outcome of several infectious diseases, such as yellow fever (YF), with the aim of benefiting human health through detection of intracellular viral antigen and vaccine efficacy trials, as well as characterization, quantification, and monitoring of immune cells subpopulations and their biological functions quality. In the last 10 years we have been facing the re-emergence of YF, considered an endemic disease caused by arbovirus in continents including South America. Unfortunately, severe forms of the disease are still associated with increased mortality. Even with the availability of effective vaccines, gaps about understanding the immune pathophysiology and clinical management are still considered a huge challenge for the scientific community. Therefore, such tool has the potential to aggregate in flavivirus setting, being effective in tracking infection in different biological culture systems, animal and human models, as well as in searching for new antiviral drugs and vaccine efficacy.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2913 ","pages":"1-17"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144044066","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

Analysis of Enzymatic Activity of Matrix Metalloproteinases by Collagen Zymography Utilizing Extracted Collagen from Bovine Muscle Tissue. 利用牛肌肉组织胶原提取，用胶原酶谱法分析基质金属蛋白酶的酶活性。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4478-2_9

Larissa A Koulicoff, Michael D Chao

引用次数: 0

Purification and Ultramicroscopic Observation of the Influenza A Virus Ribonucleoprotein Complex. 甲型流感病毒核糖核蛋白复合物的纯化及超微观察。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4326-6_7

Masahiro Nakano, Takeshi Noda

引用次数: 0

Techniques to Determine Mammalian Sperm Capacitation. 测定哺乳动物精子获能的技术。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4406-5_31

Joan E Rodríguez Gil, Olga Blanco-Prieto

{"title":"Techniques to Determine Mammalian Sperm Capacitation.","authors":"Joan E Rodríguez Gil, Olga Blanco-Prieto","doi":"10.1007/978-1-0716-4406-5_31","DOIUrl":"10.1007/978-1-0716-4406-5_31","url":null,"abstract":"The detection of the achievement of the capacitation status in a sperm sample is a very important asset for optimizing most reproductive techniques centered on semen, from freezing to \"in vitro\" fertilization. However, there is not a single, simple test that can determine the precise capacitation of a sample. This implies that a combined panel of separate tests focused on separate aspects of sperm function must be carried out to obtain a precise knowledge of the functional status of the sample. This work deals with a brief explanation of the most important techniques applied at these moments to determine sperm capacitation, with an emphasis not on the description of each technique, but on the advantages, disadvantages, and main purposes taking into account practical aspects such as the precise target by which a laboratory wants to determine capacitation. In this way, the main aim of this work is to give a practical guide for practitioners of laboratories from separate objectives, from standard semen quality analysis to molecular and/or mechanistic studies of sperm function, for choosing the most adequate tests to determine capacitation basing on the intended precise targets chosen in each case.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2897 ","pages":"463-495"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143811997","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

Quality Assurance in Metabolomics and Metabolic Profiling. 代谢组学和代谢谱的质量保证。

Methods in molecular biology Pub Date : 2025-01-01 DOI: 10.1007/978-1-0716-4334-1_2

Jennifer A Kirwan, Ulrike Bruning, Jonathan D Mosley

引用次数: 0