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GEN Protocols

Translational Medicine

Translational Medicine

Primer

Benefits of Using an Electronic Laboratory Notebook

Electronic laboratory notebooks (ELNs) were developed to keep track of experiments and data so that scientists could review their work in a single place and easily reproduce experiments. Many ELNs started out as electronic versions of paper lab notebooks. Now, the most advanced ELNs offer an extensi... More


Primer

Qualitative, Semi-quantitative, and Quantitative Immunodiagnostics for COVID-19

While qualitative reverse transcription polymerase chain reactions (RT-PCR) are used to diagnose COVID-19, serologic tests for antibodies specific to the SARS-CoV-2 virus are used to detect past infection. Most serologic assays are qualitative and semi-quantitative but quantitative serologic assays ... More


Primer

Saliva, Nasal Swabs, or Blood: Best Samples for Pathogen Detection

Most immunodiagnostic protocols and commercially available kits require serum, plasma, or blood samples. Collecting such samples requires the skills of a phlebotomist. For pathogen detection, particularly in largescale field studies, the collection of noninvasive samples such asnasal swabs, tears, o... More


Primer

Separating Blood into Cellular and Non-Cellular Components

A key diagnostic step in disease detection is analyzing blood components. Accurate separation of blood cells from plasma, which holds proteins and other constituents, is crucial in many assays.First introduced in the late 1980s, lateral flow devices, also referred to as lateral flow immunoassays, ar... More


Primer

Challenges of Organoid Culture

Complex patient models, including organoids and organ-on-a-chip devices, have been developing tremendously fast over the last decade.1 Organoids are 3D multicellular in vitro tissue constructs that functionally and genetically mimic portions of in vivo organs. Properly supplemented media allow the c... More


Primer

Ensuring Immunodiagnostic Efficacy with Pathogen Evolution

Viruses mutate frequently evolving into new variants and strains over time. Antigenic drift is the process of random genetic mutations of an infectious agent resulting in minor changes in proteins called antigens. These copying errors occur when the virus replicates. Over time, these mutations can r... More


Primer

Dried Blood Spot Testing: Advantages and Disadvantages

In vitro diagnostics, marketed directly to consumers without the involvement of a health care provider are called direct-to-consumer tests, or DTC, which are facilitated through easy-to-collect sample matrices such as dried blood spots (DBS), saliva, or swabs that allow the safe and stable transport... More


Primer

Immunodiagnostics in the Post-Covid Era

The benefits of globalization have ushered in the reality of fast-spreading infectious diseases. This has made the quick development of preventive vaccines and immune therapies imperative. Spread of pathogens through rapid and constant travel is compounded by the multipronged challenges ranging from... More


Primer

Planning Your Next Chemogenetic Experiment

Chemogenetic experiments have been used to study a wide variety of behaviors, such as triggers for substance abuse,eating disorders, obesity, obesity-associated metabolic abnormalities, long term memory, hypertension, locomotor ability, as well as pain, startle, and fear responses. For instance,Hori... More


Primer

Tips for Increasing Transfection Efficiency

Chemical transfection, which relies on the formation of a condensed complex of positively chargedliposomal or non-liposomal reagents and negatively charged nucleic acids,is the most common method for delivering foreign genetic material into cultured cells. Complexes are attracted to the negatively c... More


Protocol

Generating and maintaining uniform 3D spheroids in long term cultures

This instruction explains the basic operating procedure for generating and maintaining longterm three-dimensional spheroid cultures on PrimeSurface® 96 Slit-well plates. Instructions recommended here are the best operating protocols but depending upon your cell line and assay, some optimizations may... More


Protocol

Preparing a Brain-Mimetic 3D Matrix for Culturing Human Pluripotent Stem Cells

This is a protocol for culturing human pluripotent stem cells (hPSCs) using a 3D matrix. It provides instructions for preparing a brain-mimetic 3D scaffold that can support neuronal growth and for culturing hPSCs. This injectable scaffold can be potentially employed in the field of regenerative medi... More


Protocol

Detergent-Free Nuclei Isolation

A novel spin column-based approach to isolate intact single nuclei from tissues (fresh or frozen) and cells without using a detergent. The protocol is fast, easy, and reliable, producing high-quality single nuclei with minimized clumping and possible leakage of the nuclear envelope. The protocol is ... More


Protocol

Short Communication: Ischemic Stroke Subtypes Among Thai HIV-Infected Patients: A 12-Year Retrospective Study

Traditional vascular risk factors and HIV-related factors can contribute to higher prevalence of ischemic stroke (IS) among HIV-infected patients. Understanding the underlying mechanism of IS could lead to proper management and prognosis. We aimed to determine the prevalence of IS subtypes among HIV... More


Protocol

Short Communication: HIV Viral Load Trends During the Coronavirus Disease 2019 Pandemic in a Reference Center for HIV in Rome, Italy

Coronavirus disease 2019 (COVID-19) pandemic has reduced the access of HIV patients to reference centers. However, retention-in-care is critical to maintain adherence to therapy and viral suppression. During lockdown in Italy, our center implemented several measures to ensure HIV-care continuum. To ... More


Protocol

Short Communication: Communication Between Pregnant Women and Male Partners About HIV Testing in the United States

Male partner uptake of HIV testing during antenatal care is poor despite women's reported desire for partner testing. This qualitative study of HIV-negative pregnant women and their partners in a high HIV prevalence city in the United States assessed communication between partners about HIV testing.... More


Protocol

Short Communication: Efficacy and Safety of Dolutegravir Plus Lamivudine as a First-Line Regimen in Clinical Practice

The GEMINI trials have showed that the two drugs regimen of dolutegravir+lamivudine (DTG +3TC) was noninferior to a three-drug regimen as a first line regimen for treatment-naive people living with HIV. The aim of our study was to confirm, in a real-life setting, the efficacy of this regimen. We con... More


Protocol

Short Communication: Update in Natural Antiretroviral Resistance-Associated Mutations Among HIV Type 2 Variants and Discrepancies Across HIV Type 2 Resistance Interpretation Tools

HIV variants carry natural polymorphisms related to drug resistance (R-markers) fixed during viral evolution in the absence of antiretroviral therapy (ART) that may impact on drug susceptibility and resistance pathways. We aimed to identify the HIV type 2 (HIV-2) variant-specific R-markers at Pol in... More


Protocol

Short Communication: Coronavirus Disease 19 Among People Living with HIV in Western India: An Observational Cohort Study

A retrospective cohort study was conducted to assess clinical characteristics and outcomes of coronavirus disease-19 (COVID-19) among people living with HIV (PLHIV) in western India. Out of 86 PLHIV with COVID-19 illness, 19.7% had severe/critical illness and 6 (6.9%) individuals died. Median (inter... More


Protocol

Short Communication: Higher Tenofovir Concentrations in Hair Are Associated with Decreases in Viral Load and Not Self-Reported Adherence in HIV-Infected Adolescents with Second-Line Virological Treatment Failure

Objective methods of measuring antiretroviral adherence are limited. We assessed the relationship between tenofovir disoproxil fumarate (TDF) hair concentrations, self-reported adherence, and virological outcomes in HIV-infected adolescents in Harare, Zimbabwe. HIV-infected adolescents on atazanavir... More


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Primer

Two Critical Steps to Success in Multicolor Flow Cytometry

Flow cytometry is a powerful tool to analyze and characterize cells based on their protein expression, most often those expressed on the cell surface. Multicolor flow cytometry uses multiple fluorochromes as primary or secondary antibody conjugates to label cells. The capacity for modern flow cytome... More


Primer

Expressing, Extracting and Purifying Membrane Proteins

Membrane proteins are key biological players involved in myriad essential cellular processes from molecular transport, cellular communication, and signal transduction to maintaining structures within cells and tissues. Despite their importance, accounting for 25% of all proteins in the human genome ... More


Primer

Unwanted Fungi: Detecting Microbial Cell Culture Contamination

Coming into the lab and spotting an obvious fungal growth, or the tell-tale signs of cloudy media, can be a stressful experience for researchers. When carrying out cell culture in the lab, contamination from microbial agents such as bacteria and fungus can occur and have disastrous effects on ongoin... More


Primer

Source and Elimination of Common Mass Spectrometry Contaminants

Sensitivity is a key attribute of mass spectrometers to allow quantification of minute amounts of analytes. Yet this sensitivity can be a conundrum. Myriad sources can contribute contaminants that can impact background and confound interpretation of results. An increased background signal can make i... More


Primer

Assessing qRT-PCR COVID-19 Testing Results: What Can Go Wrong?

COVID-19 is commonly diagnosed by quantitative fluorescence real-time reverse transcriptase-polymerase chain reaction (qRT-PCR) (1, 2). The test is the sequential action of two enzymes--an RNA-dependent DNA polymerase, also known as a reverse transcriptase, to copy the SARS-CoV-2 RNA into complement... More


Primer

Pathogen Detection Assays: Advantages, Disadvantages and Alternatives

Viral infections are difficult to diagnose based on the patient’s clinical presentation alone. A reliable and accurate diagnosis requires laboratory-based testing for the pathogen of interest. Such laboratory testing involves either the direct detection of viral antigens, or detection of virus-speci... More


Primer

Benefits of Using an Electronic Laboratory Notebook

Electronic laboratory notebooks (ELNs) were developed to keep track of experiments and data so that scientists could review their work in a single place and easily reproduce experiments. Many ELNs started out as electronic versions of paper lab notebooks. Now, the most advanced ELNs offer an extensi... More


Primer

Qualitative, Semi-quantitative, and Quantitative Immunodiagnostics for COVID-19

While qualitative reverse transcription polymerase chain reactions (RT-PCR) are used to diagnose COVID-19, serologic tests for antibodies specific to the SARS-CoV-2 virus are used to detect past infection. Most serologic assays are qualitative and semi-quantitative but quantitative serologic assays ... More


Primer

Saliva, Nasal Swabs, or Blood: Best Samples for Pathogen Detection

Most immunodiagnostic protocols and commercially available kits require serum, plasma, or blood samples. Collecting such samples requires the skills of a phlebotomist. For pathogen detection, particularly in largescale field studies, the collection of noninvasive samples such asnasal swabs, tears, o... More


Primer

Separating Blood into Cellular and Non-Cellular Components

A key diagnostic step in disease detection is analyzing blood components. Accurate separation of blood cells from plasma, which holds proteins and other constituents, is crucial in many assays.First introduced in the late 1980s, lateral flow devices, also referred to as lateral flow immunoassays, ar... More


Mixer

Organoids for Airway and Alveolar Disease Modeling

In this Mixer, we will discuss the development of respiratory airway and alveolar organoids derived from human iPSCs and how these are used in modeling respiratory diseases such as cystic fibrosis. Airway organoid models can be used for testing the therapeutic efficacy of candidate drugs for respira... More


Video

GEN Protocols Expert Exchanges: Tools and Techniques to Develop Therapies for Neurological Disorders

The human brain is an exquisitely complex organ, consisting of nearly one hundred billion neurons, an equivalent number of non-neuronal cells and trillions of synapses. Understanding the factors that play a role in the development of the structure and function of the brain is an essential step to de... More


Video

GEN Protocols Expert Exchanges: Patient-Derived Pancreatic Cancer Organoids Predict Response to Therapy

Pancreatic cancer accounts forthree percentof all cancers in the United States and is responsible for seven percent of all cancer deaths. Miniature organoids cloned from pancreatic tumors in patients are being grown in laboratories to conduct assays that can predict the effectiveness of chemotherapy... More


Video

GEN Protocols Expert Exchanges: Critical ALS Biomarkers of Neuroinflammation & Oxidative Stress

Biomarkers are essential tools for the development of therapeutics, clinical trial design and patient stratification. Biomarkers also help evaluate the efficacy of therapeutic intervention on disease progression or relapse. Yet, biomarker driven patient stratification, a hallmark in oncology trials,... More


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