Immunization of MERS-CoV-2-infected mice with a sublethal dose of MERS-CoV or VRP-MERS-S.

Here, we detail the immunization of mice with a sublethal dose of MERS-CoV or two doses of replication-incompetent alphavirus replicon particles expressing MERS-CoV spike protein. We then describe steps to determine the outcome of immunization by challenging immunized mice with a lethal dose of MERS-CoV, as well as by detecting virus-specific neutralizing antibody and virus-specific T cell response via neutralization assay and flow cytometry, respectively. This protocol can be used to evaluate other CoV infections or vaccine-induced immune responses. For complete details on the use and execution of this protocol, please refer to Zheng et al. (2021).1.

Interdisciplinary approach to biomedical research: a panacea to efficient research output during the global pandemic

Biomedical research is rapidly growing due to inventions and developments in science and technology. Several interdisciplinary fields should be combined to find the remedy of diseases including pandemics. To accomplish this, interdisciplinary research is a prerequisite. Using improved techniques in microscopy and genetic engineering, the systemic perspective of the human body and related diseases can be found. Recent genetic-based inheritance studies of diseases, understanding various omics, stem cell systems, and gene editing tools including CRISPR relevant to biomedical research require multidisciplinary approach. Improvements in the field of bioinformatics and efficient use of model organisms in clinical testing including drug assessment are important disciplines common in different researches. The merging of different closely related areas of medical research will produce suitable changes in diagnosis and treatment. In the present scenario of increased global pandemic hits like COVID-19, an understanding on the interdisciplinary approach is needed for controlling the spread and finding vaccines. © 2022 Elsevier Inc. All rights reserved.

When should researchers cite study differences in response to a failure to replicate?

Scientists often respond to failures to replicate by citing differences between the experimental components of an original study and those of its attempted replication. In this paper, we investigate these purported mismatch explanations. We assess a body of failures to replicate in neuroscience studies on spinal cord injury. We argue that a defensible mismatch explanation is one where (1) a mismatch of components is a difference maker for a mismatch of outcomes, and (2) the components are relevantly different in the follow-up study, given the scope of the original study. With this account, we argue that not all differences between studies are meaningful, even if they are difference makers. As our examples show, focusing only on these differences results in disregarding the representativeness of the original experiment's components and the scope of its outcomes, undercutting other epistemic aims, such as translation, in the process.

Deep Metabolic Profiling Assessment of Tissue Extraction Protocols for Three Model Organisms.

Metabolic profiling harbors the potential to better understand various disease entities such as cancer, diabetes, Alzheimer's, Parkinson's disease or COVID-19. To better understand such diseases and their intricate metabolic pathways in human studies, model animals are regularly used. There, standardized rearing conditions and uniform sampling strategies are prerequisites towards a successful metabolomic study that can be achieved through model organisms. Although metabolomic approaches have been employed on model organisms before, no systematic assessment of different conditions to optimize metabolite extraction across several organisms and sample types has been conducted. We address this issue using a highly standardized metabolic profiling assay analyzing 630 metabolites across three commonly used model organisms (Drosophila, mouse, and zebrafish) to find an optimal extraction protocol for various matrices. Focusing on parameters such as metabolite coverage, concentration and variance between replicates we compared seven extraction protocols. We found that the application of a combination of 75% ethanol and methyl tertiary-butyl ether (MTBE), while not producing the broadest coverage and highest concentrations, was the most reproducible extraction protocol. We were able to determine up to 530 metabolites in mouse kidney samples, 509 in mouse liver, 422 in zebrafish and 388 in Drosophila and discovered a core overlap of 261 metabolites in these four matrices. To enable other scientists to search for the most suitable extraction protocol in their experimental context and interact with this comprehensive data, we have integrated our data set in the open-source shiny app "MetaboExtract". Hereby, scientists can search for metabolites or compound classes of interest, compare them across the different tested extraction protocols and sample types as well as find reference concentration values.

Staphylococcus massiliensis isolated from human blood cultures, Germany, 2017-2020.

Clinical and laboratory data on newly described staphylococcal species is rare, which hampers decision-making when such pathogens are detected in clinical specimens. Here, we describe Staphylococcus massiliensis detected in three patients at a university hospital in southwest Germany. We report the discrepancy of microbiological findings between matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, 16S-rRNA polymerase chain reaction, and whole-genome sequencing for all three isolates. Our findings highlight the diagnostic pitfalls pertinent to novel and non-model organisms in daily microbiological practice, in whom the correct identification is dependent on database accuracy.

Optimized immunofluorescence staining protocol for imaging germinal centers in secondary lymphoid tissues of vaccinated mice.

Location of immune cells that form the germinal center reaction within secondary lymphoid tissues can be characterized using confocal microscopy. Here, we present an optimized immunofluorescence staining protocol to image germinal center structures in fixed/frozen spleen sections from ChAdOx1 nCoV-19 immunized mice. This protocol can be adapted to identify other cell types within secondary lymphoid tissues. For complete information on the generation and use of this protocol to examine immune responses to the COVID vaccine ChAdOx1 nCoV-19, please refer to Silva-Cayetano et al. (2020).

A qualitative IgG ELISA for detection of SARS-CoV-2-specific antibodies in Syrian hamster serum samples.

This protocol describes an indirect enzyme-linked immunosorbent assay for qualitative detection of IgG antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in Syrian hamster serum samples. We describe the preparation of inactivated virus antigens and the negative control antigen and the use of antigen-coated microtiter plates to detect SARS-CoV-2-specific antibodies from SARS-CoV-2-infected hamsters, including the criteria for differentiating positive versus negative reaction. The limited batch-to-batch variability of this assay has been verified with two batches of independently prepared antigens. For complete details on the use and execution of this protocol, please refer to Mohandas et al. (2021).

Medical toolkit organisms and Covid-19.

The Covid-19 pandemic has intensified interest in animals with superior antiviral defences. I argue that the role of such animals in biomedical research contrasts with the role of disease models.

Sensitization of Non-permissive Laboratory Mice to SARS-CoV-2 with a Replication-Deficient Adenovirus Expressing Human ACE2.

Common laboratory mice such as BALB/c and C57BL/6 mice are not permissive to SARS-CoV2 infection. Sensitization of laboratory mice with Adenovirus expressing human ACE2 (Ad5-hACE2) provides a rapid model for testing viral intervention in vivo. Despite the lack of lethal outcome, Ad5-hACE2-sensitized mice show 20% weight loss on average upon viral challenge with infectious virus being detected at the site of sensitization. This protocol describes the sensitization and subsequent infection of common laboratory mice for use in testing anti-viral interventions. For complete details on the use and execution of this protocol, please refer to Sun et al. (2020).