Biology subjects

Edwards, E. E.

Publications and source records attributed to Edwards, E. E..

2 recordsLinked to original sources

bioRxiv2024

From birds to mammals: spillover of highly pathogenic avian influenza H5N1 virus to dairy cattle led to efficient intra- and interspecies transmission

Infections with the highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.4.4b virus have resulted in the death of millions of domestic birds and thousands of wild birds in the U.S. since January, 20221-4 Throughout this outbreak, spillovers of the virus to mammals have been frequently documented5-12. Here, we report the detection of HPAI H5N1 virus in dairy cattle herds across several states in the U.S. The affected cows displayed clinical signs encompassing decreased feed intake, altered fecal consistency, respiratory distress, and decreased milk production with abnormal milk. Infectious virus and RNA were consistently detected in milk collected from affected cows. Viral staining in tissues revealed a distinct tropism of the virus for the epithelial cells lining the alveoli of the mammary gland in cows. Analysis of whole genome sequences obtained from dairy cows, birds, domestic cats, and a racoon from affected farms indicated multidirectional interspecies transmissions. Epidemiologic and genomic data revealed efficient cow-to-cow transmission after healthy cows from an affected farm were transported to a premise in a different state. These results demonstrate the transmission of HPAI H5N1 clade 2.3.4.4b virus at a non-traditional interface and to a new and highly relevant livestock species, underscoring the ability of the virus to cross species barriers.

microbiology↗

bioRxiv2022

A Scalable 3D High-Content Imaging Protocol for Measuring a Drug Induced DNA Damage Response Using Immunofluorescent Sub-nuclear γH2AX Spots in Patient Derived Ovarian Cancer Organoids

The high morbidity rate of ovarian cancer has remained unchanged during the past four decades, partly due to lack of understanding of disease mechanisms and difficulties in developing new targeted therapies. Defective DNA damage detection and repair is one of the hallmarks of cancer cells and is a defining characteristic of ovarian cancer. Most in vitro studies to date, involve viability measurements at scale using relevant cancer cell lines, however, the translation to clinic is often lacking. The use of patient derived organoids is closing that translational gap yet the 3D nature of organoid cultures present challenges for assay measurements beyond viability measurements. In particular, high-content imaging has the potential for screening at scale providing a better understanding of mechanism of action of drugs or genetic perturbagens. In this study we report a semi-automated and scalable immunofluorescence imaging assay utilising the development of a 384-well plate based subnuclear staining and clearing protocol and optimisation of 3D confocal image analysis for studying DNA damage dose response in human ovarian cancer organoids. The assay was validated in four organoid models and demonstrated a predictable response to Etoposide drug treatment with lowest efficacy observed in the clinically most resistant model. This imaging and analysis method can be applied to other 3D organoid and spheroid models for use in high content screening.

bioinformatics↗