The Production and characterization of Polyclonal Antibodies Against Interferon Alpha in Mice

The polyclonal antibodies are used extensively for research purposes in many areas of biology, such as immunoprecipitation, histochemistry, enzyme linked immunosorbent assays (ELISA), diagnosis of disease and western blots. Typically, an animal’s immune system will generate a large group of antibodies that recognize several epitopes of a particular antigen. Interferon alpha plays an important role in immune response activation and therefore is of interest in studies related to autoimmune diseases. In this paper the production of antibodies against interferon was studied in order to quantify interferon production to analyze interferon levels in autoimmune disorders in the future. For the antibody production, one month old laboratory grade mice were injected with interferon alpha in combination with a Freund’s complete adjuvant for a course of five weeks after which the antibodies were obtained in mouse serum. Confirmation of the production of anti-interferon alpha antibodies was carried out by the Elisa, immune dot blot and western blot analysis. An interferon alpha of approximately 20.5-21.5 KDa was detected in immunedot blot test. These antibodies may be produced in these mouse models commercially and could be used in future for treatment of autoimmune diseases by managing the interferon levels in the patients. Copyright(c) The Authors

How the Innate Immune System Fights for Your Health

When your body encounters intruders like viruses, bacteria, fungi, or parasites, this invasion triggers a complex and amazing process called the immune response. Activation of the body’s immune system is necessary to fight off these intruders, but it must also distinguish them from the body’s own healthy tissues. The goal of the immune response is to keep the body healthy. The earliest responses that occur to protect the body from invading organisms is called the innate immune response. In this article, we explain the components of the innate immune system and how this system helps to keep the body safe from dangerous invaders.

Impact of Different Waves of COVID-19 on Emergency Medical Services and Out-of-hospital Cardiopulmonary Arrest in Madrid, Spain

Background: COVID-19 has led to decreased survival of out-of-hospital cardiorespiratory arrest (OHCA). We analysed the impact of the first COVID-19 pandemic year on emergency medical services and OHCA care compared with the previous year.Methods: Data for this observational study were collected for OHCAs attended by the SUMMA 112 emergency service during March 2019 to March 2021. We compared data covering 15 March 2020–14 March 2021 (pandemic year) to retrospective data covering 15 March 2019–14 March 2020 (non-pandemic year). Results: During the pandemic period, 1743 OHCA patients were attended, compared to 1781 during the non-pandemic year. Median patient age during the pandemic period was lower than in the non-pandemic period (71 vs 72, p=0·037). Emergency services response activation time increased during the pandemic year, to 3 minutes, 16 seconds from 2 minutes, 48 seconds in the non-pandemic period (p=0·001). Time to arrival at the scene also increased during the pandemic (12 minutes vs 11 minutes, 25 seconds before the pandemic; p=0·001). The percentage of OHCAs in which resuscitation was attempted was lower during the pandemic (59·4% vs 62·9%, p=0·034), as were survival on hospital arrival (30·3% vs 34·6%, p=0·04). Differences in response activation time (p=0·003) and scene arrival times (p=0·003) were greater during the first pandemic wave compared with the later phases. Conclusions: The different phases of the pandemic variably affected OHCA care. The first wave led to longer resource activation, increased home events and scene arrival times, as well as lower patient survival.

Transcriptomic mapping of the inter-individual variability of cellular stress response activation in primary human hepatocytes

Background & Aims: One of the early key events of drug-induced liver injury (DILI) is the activation of adaptive stress responses, a cellular mechanism to overcome stress. Given the diversity of DILI outcomes and lack in understanding of population variability, we mapped the inter-individual variability in stress response activation to improve DILI prediction. Approach & Results: High-throughput transcriptome analysis of over 8,000 samples was performed in primary human hepatocytes of 50 individuals upon 8 to 24 h exposure to broad concentration ranges of stress inducers: tunicamycin to induce the unfolded protein response (UPR), diethyl maleate for the oxidative stress response, cisplatin for the DNA damage response and TNFα for NF-κB signalling. This allowed investigation of the inter-individual variability in concentration-dependent stress response activation, where the average of benchmark concentrations (BMCs) had a maximum difference of 864, 13, 13 and 259-fold between different hepatocytes for UPR, oxidative stress, DNA damage and NF-κB signalling-related genes, respectively. Hepatocytes from patients with liver disease resulted in less stress response activation. Using a population mixed-effect framework, the distribution of BMCs and maximum fold change were modelled, allowing simulation of smaller or larger PHH panel sizes. Small panel sizes systematically under-estimated the variance and resulted in low probabilities in estimating the correct variance for the human population. Moreover, estimated toxicodynamic variability factors were up to 2-fold higher than the standard uncertainty factor of 101/2 to account for population variability during risk assessment, exemplifying the need of data-driven variability factors. Conclusions: Overall, by combining high-throughput transcriptome analysis and population modelling, improved understanding of variability in stress response activation across the human population could be established, thereby contributing towards improved prediction of DILI.

NAD+ Degrading Enzymes, Evidence for Roles During Infection

Declines in cellular nicotinamide adenine dinucleotide (NAD) contribute to metabolic dysfunction, increase susceptibility to disease, and occur as a result of pathogenic infection. The enzymatic cleavage of NAD+ transfers ADP-ribose (ADPr) to substrate proteins generating mono-ADP-ribose (MAR), poly-ADP-ribose (PAR) or O-acetyl-ADP-ribose (OAADPr). These important post-translational modifications have roles in both immune response activation and the advancement of infection. In particular, emergent data show viral infection stimulates activation of poly (ADP-ribose) polymerase (PARP) mediated NAD+ depletion and stimulates hydrolysis of existing ADP-ribosylation modifications. These studies are important for us to better understand the value of NAD+ maintenance upon the biology of infection. This review focuses specifically upon the NAD+ utilising enzymes, discusses existing knowledge surrounding their roles in infection, their NAD+ depletion capability and their influence within pathogenic infection.