Bondehoffmann5605

Machine learning is being employed for the development of diagnostic methods for several diseases, but prognostic techniques are still poorly explored. The development of such approaches is essential to assist healthcare workers to ensure the most appropriate treatment for patients. In this chapter, we demonstrate a detailed protocol for the application of machine learning to MALDI-TOF MS spectra of COVID-19-infected plasma samples for risk classification and biomarker identification.The use of in vitro methods of infecting cell lines to test new treatments for SARS-CoV-2 does not always recapitulate the real context of the infection, and mouse models for SARS-CoV-2 infection are limited. Here we describe a novel ex vivo approach by collecting, isolating, and culturing nasal epithelial cells obtained from patients with COVID-19. This technique allows us to study immune responses and test new treatments directly on cells from patients naturally infected with SARS-CoV-2.Coenzyme Q10 (CoQ10) plays an essential electron carrier role in the mitochondrial electron transfer chain (ETC) as well as being a potent antioxidant and influencing inflammatory mediators. In view of these functions, the reason why certain individuals may be more susceptible to the severe disease or long-term complications (long COVID) of COVID-19 infection may be associated with an underlying deficit in cellular CoQ10 status. Thus, our group has outlined an analytical method for the determination of cellular CoQ10 status using HPLC linked UV detection at 275 nm. This method has been utilized in patient tissue samples to investigate evidence of a CoQ10 deficiency and thus may have potential in determining the possible susceptibility of individuals to severe disease associated with COVID-19 infection or to long COVID.The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent driving the current COVID-19 pandemic. The acute respiratory distress that occurs in some severe COVID-19 cases has been linked with hypercoagulation or thrombotic events as well as a worse prognosis and increased risk of death. Thus, point-of-care devices that can be used for early detection of coagulation abnormalities would assist in COVID-19 management. This chapter describes the use of the Roche Diagnostics CoaguChek® XS test kit for potential use in COVID-19 personalized medicine approaches.Infection with SARS-CoV-2, the causative agent of COVID-19, causes numerous cellular dysfunctions. The virus enters the host cells and hijacks the cell machinery for its replication, resulting in disturbances of the oxidative, reductive balance, increased production of damaging reactive oxygen species (ROS), and mitochondrial dysfunction. This damaging cycle can make cells less resistant to infection and make the host more likely to experience a severe disease course. Treatment with antioxidants has been tested as a potential approach to reduce the effects of this disorder. Here, we present a protocol to assess the impact of treatment with a mixture of curcuminoids on physiological and molecular biomarkers, focusing on determining total antioxidant capacity. We used a cohort of diabetes patients with an imbalance in redox mechanisms as such patients are more likely to become severely ill from COVID-19 than healthy persons.Inflammatory diseases caused by infectious agents such as the SARS-CoV-2 virus can lead to impaired reductive-oxidative (REDOX) balance and disrupted mitochondrial function. Peripheral blood mononuclear cells (PBMCs) provide a useful model for studying the effects of inflammatory diseases on mitochondrial function but can be limited by the need to store these cells by cryopreservation prior to assay. Here, we describe a method for improving and determining PBMC viability with normalization of values to number of living cells. The approach can be applied not only to PBMC samples derived from patients with diseases marked by an altered inflammatory response such as viral infections.Severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) has been identified as the causative agent of COVID-19. Accurate detection of SARS-CoV-2 infection is not only important for management of infected individuals but also to break the chain of transmission. Although the polymerase chain reaction (PCR) is the gold standard for diagnosis of acute SARS-CoV-2 infection, there are a number of limitations of these assays, which include the inability to detect past infection and decline in sensitivity 14 days post-symptom onset. There are several serology tests developed for the detection of SARS-CoV-2 antibodies including high-throughput serology platforms and lateral flow immunoassays. These tests should be evaluated for their performance to meet local regulations acceptance criteria. To optimize the diagnostic algorithm for SARS-CoV-2, this protocol describes the evaluation of serological antibody testing using various automated serology platforms and lateral flow immunoassays. This protocol was evaluated in both serum and plasma samples. The sample preparation, procedure, and data analysis are described. The protocol can be adapted for any serological testing.Approximately one in three people infected with the SARS-CoV-2 virus have mild symptoms or are asymptomatic. However, these individuals can still spread the virus. Regular self-testing can help to detect these individuals and thereby slow the spread and protect the more vulnerable members of society. Here, we present a protocol for use of the COVID-19 rapid antigen test which was made freely available to residents of the United Kingdom in April of this year. This using the lateral flow technique for detection of antigens and is amenable to multiplexing.SARS-CoV-2 can stimulate the expression of various inflammatory cytokines and induce the cytokine storm in COVID-19 patients leading to multiple organ failure and death. Curcumin as a polyphenolic compound has been shown to have anti-inflammatory properties and inhibit the release of numerous pro-inflammatory cytokines. We present multiplex analysis using the Evidence Investigator biochip system to determine the effect of curcumin on serum level of cytokines which are typically elevated in cytokine storm events, including tumor necrosis factor (TNF-α), interleukin 6 (IL-6), and IL-10.Statins have pleiotropic effects on inflammatory responses in addition to their lipid-lowering action, which contributes to their favorable effect on cardiovascular disorders. Statins affect adhesion, migration, antigen presentation, and cytokine generation of immune cells. Pre-clinical and clinical studies suggest that statin intervention targeted early in the infection might help COVID-19 patients to reduce the effects of acute respiratory distress syndrome (ARDS), the cytokine storm, and vascular collapse by modulating harmful pathogenic mechanisms. This chapter presents a protocol for measuring blood-based biomarkers predictive of these responses in COVID-19 patients using two specific multiplex immunoassays that target proteins that differ widely in concentration.Serological assays have been a useful tool for detection of antibodies to SARS-CoV-2 during the COVID-19 pandemic. These assays are used for epidemiology and serosurveillance to monitor the progression of the pandemic, to identify and differentiate individuals who have developed antibodies from natural infection versus vaccine-induced immunity, and to identify potential donors of convalescent plasma for therapeutic purposes. In this chapter, we describe a commercially available bead-based serological assay, the Luminex® xMAP® SARS-CoV-2 Multi-Antigen IgG Assay, that detects and identifies antibodies against three SARS-CoV-2 antigens. In addition to the assay principle and workflow, we describe modifications that may be used to evaluate alternate sample types, antibody isotypes, and potential neutralizing antibody responses.Severe cases of SARS-CoV-2 and other pathogenic virus infections are often associated with the uncontrolled release of proinflammatory cytokines, known as a "cytokine storm." We present a protocol for multiplex analysis of three cytokines, tumor necrosis factor-alpha (TNF-a), interleukin 6 (IL-6), and IL-10, which are typically elevated in cytokine storm events and may be used as a predictive biomarker profile of disease severity or disease course.Most people infected by the SARS-CoV-2 virus which causes COVID-19 disease experience mild or no symptoms. Severe forms of the disease are often marked by a hyper-inflammatory response known as a cytokine storm. Thus, biomarker tests which can identify these patients and place them on the appropriate treatment regime at the earliest possible phase would help to improve outcomes. Here we describe an automated microarray-based immunoassay using the Fraunhofer lab-on-a-chip platform for analysis of C-reactive protein due to its role in the hyper-inflammatory response.Since the emergence of COVID-19, concerted worldwide efforts have taken place to minimize global spread of the contagion. Its widespread effects have also facilitated evolution of new strains, such as the delta and omicron variants, which emerged toward the end of 2020 and 2021, respectively. While these variants appear to be no more deadly than the previous alpha, beta, and gamma strains, and widespread population vaccinations notwithstanding, greater virulence makes the challenge of minimizing spread even greater. One of the peculiarities of this virus is the extreme heath impacts, with the great majority of individuals minimally affected, even sometimes unaware of infection, while for a small minority, it is deadly or produces diverse long-term effects. Apart from vaccination, another approach has been an attempt to identify treatments, for those individuals for whom the virus represents a threat of particularly severe health impact(s). These treatments include anti-SARS-CoV-2 monoclonal antibodies, anticoagulant therapies, interleukin inhibitors, and anti-viral agents such as remdesivir. Nutritional factors are also under consideration, and a variety of clinical trials are showing promise for the use of specific fatty acids, or related compounds such as fat-soluble steroid vitamin D, to mitigate the more lethal aspects of COVID-19 by modulating inflammation and by anti-viral effects. Here we explore the potential protective role of fatty acids as a potential prophylactic as well as remedial treatment during viral infections, particularly COVID-19. We present a multiplexed method for the analysis of free and phospholipid bound fatty acids, which may facilitate research into the role of fatty acids as plasma biomarkers and therapeutic agents in minimizing pre- and post-infection health impacts.COVID-19 disease is caused by infection with the SARS-CoV-2 virus and is associated with a cytokine storm effect in some patients. This can lead to decreased ability of the host to cope with the infection and result in severe disease outcomes. Here, we present a protocol for isolation of peripheral blood mononuclear cells (PBMCs) from COVID-19 patients followed by liquid chromatography-mass spectrometry (LC-MS) profiling to identify the affected molecules and molecular pathways. It is hoped that this will lead to the identification of potential biomarkers for monitoring the disease as well as treatment responses. Selleck SCH-527123 This approach could also be used in the study of other respiratory viruses.