Methods in molecular biology
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Coronaviruses (CoVs), enveloped positive-sense RNA viruses, are characterized by club-like spikes that project from their surface, an unusually large RNA genome, and a unique replication strategy. CoVs cause a variety of diseases in mammals and birds ranging from enteritis in cows and pigs, and upper respiratory tract and kidney disease in chickens to lethal human respiratory infections. Most recently, the novel coronavirus, SARS-CoV-2, which was first identified in Wuhan, China in December 2019, is the cause of a catastrophic pandemic, COVID-19, with more than 8 million infections diagnosed worldwide by mid-June 2020. Here we provide a brief introduction to CoVs discussing their replication, pathogenicity, and current prevention and treatment strategies. We will also discuss the outbreaks of the highly pathogenic Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) and Middle Eastern Respiratory Syndrome Coronavirus (MERS-CoV), which are relevant for understanding COVID-19.
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Measuring the Replicative Lifespan of Saccharomyces cerevisiae Using the HYAA Microfluidic Platform.
The replicative aging of the budding yeast, Saccharomyces cerevisiae, has been a useful model for dissecting the molecular mechanisms of the aging process. Traditionally, the replicative lifespan (RLS) is measured by manually dissecting mother cells from daughter cells, which is a very tedious process. Since 2012, several microfluidic systems have been developed to automate the dissection process, significantly accelerating RLS determination. Here, we describe a detailed protocol of RLS measurement using a ommercially available microfluidic system based on the HYAA chip design, which enables data collection of up to 8000 cells in a single experiment.
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Esophageal squamous cell carcinoma (ESCC) is an aggressive disease. Many patients have locally advanced disease or already have distant metastasis at presentation. ⋯ Definitive radiotherapy (RT) with or without chemotherapy is used in patients who decline surgery or are medically inoperable. Palliative radiotherapy using external beam radiotherapy or intraluminal brachytherapy is effective for dysphagia and pain control.d.
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Methylation of cytosines in DNA is the most stable type of epigenetic modification that is established and maintained by different enzymes. In plants, DNA methylation is inherited from one generation to another leaving an epigenetic mark as a memory of previous state, which may include encounter with stress or pathogen. ⋯ Though the sequencing technologies have evolved drastically, analysis of WGBS data still remains challenging. Here, we provide a methodology for performing WGBS data analysis along with critical steps for identification of methylation marks in plant genomes including legumes.
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Biobanking is important and fundamental for research and personalized medicine in patients with esophageal squamous cell carcinoma. The process often involves prospective collection of surgically obtained tissues (tissue banking) as well as serial blood samples (liquid biopsies) from the patients with esophageal squamous cell carcinoma. Apart from frozen tissues, formalin-fixed paraffin-embedded tissues are important sources of translational research. ⋯ The success and sustainability of the process needs proper infrastructure, advanced planning, funding, and multidisciplinary collaborations. The understanding of the principles and issues are detrimental for the success of biobanking. The technical procedures involved are standardized, complex, and time-consuming and needs coordinated taskforce.