Heterogenous pattern of the inflammasome-related markers in the critically ill COVID-19 patients at the ICU admission.

Purpose: Development of targeted biological therapies to COVID-19 requires reliable biomarkers that could help to indicate the responding patients. Hyperactivation of the inflammasome by SARS-CoV2 virus is hypothesized to contribute to severe course of the COVID-19 disease. Therefore, we aimed to evaluate the prognostic value of several inflammasome-related cytokines and proteins at the admission to the intensive care unit. Patients and methods: Plasma samples were obtained from 45 critically-COVID-19 patients and from 10 patients with severe craniocerebral traumatic brain injury (TBI) at the admission to the ICU. The concentration of IL-1𝛼, IL-1𝛽, IL-18, IL-1RA, galectin-1, ASC, LDH, ferritin and gasdermin D were analyzed. A novel cell-free caspase-1 plasma assay was developed by inhibitor-based immunoprecipitation followed by Western Blot. Demographic and clinical characteristics were recorded. Results: The inflammasome-related biomarkers were in similar concentration in COVID-19 and TBI patients except for galectin-1 being lower in the former. None of the tested markers was related to the outcome, length of stay or development of secondary infections. Patients with SOFA score of >9 at admission who were at high risk of death had significantly higher galectin-1 but lower IL-1RA in comparison to low-risk patients. Weak but significant correlations were observed for IL-1𝛼 and IL-1𝛽 and platelets and also for ferritin and INR. Activated caspase-1 p35 was detectable in 12/22 COVID-19 patients and was related with higher fibrinogen and lower D-dimers. It was also significantly higher in patients with SOFA>9. Conclusion: Our results indicate that the activation of the inflammasome in critically ill COVID-19 patients is a heterogenous process and is not directly related with outcome. Therefore, potential interventions aimed at this pathway in this group of patients can be limited and should be biomarker-guided.

Cross-validation of a high-performance liquid chromatography nevirapine plasma assay in a resource-limited setting in Zimbabwe

An international HIV pharmacology specialty laboratory (PSL) was established at the University of Zimbabwe to increase bioanalytical and investigator capacities. Quantitation of plasma nevirapine in samples from the AIDS Clinical Trials Group protocol 5279 was compared between the University of Nebraska Medical Center PSL and the University of Zimbabwe PSL. Both PSLs employed internally developed methods utilising reverse-phase high-performance liquid chromatography with ultraviolet detection. Eighty-seven percent of the cross-validation results exhibited ± 20% difference.

Clinical application of volumetric absorptive microsampling to the gefapixant development program

In this paper we show the application of the Tasso OnDemand™, a novel automated sample collection device, in conjunction with volumetric absorptive microsampling (VAMS) for the development of gefapixant, a P2X3 receptor antagonist currently under clinical development for the treatment of refractory and unexplained chronic cough and endometriosis-related pain. A LC–MS/MS bioanalytical method was developed and validated using VAMS to support this development program. This method was utilized in a drug–drug interaction study to establish a mathematical bridging relationship with data obtained from a validated plasma assay used to support the program. The VAMS bioanalytical method and the predictability of the mathematical relationship is reported and discussed here.

Human Immunodeficiency Virus (HIV)-Antibody Repertoire Estimates Reservoir Size and Time of Antiretroviral Therapy Initiation in Virally Suppressed Perinatally HIV-Infected Children

Different specific antibody responses against 10 HIV-1 viral proteins detected by Western blot, plasma assay on a very small amount of plasma (20 μL) can estimate HIV-DNA size and timing of ART initiation in long-term virally suppressed children.