Anaerobic microbes play crucial roles in environmental processes, industry, and human health. Traditional methods for monitoring the growth of anaerobes, including plate counts or subsampling broth cultures for optical density measurements, are time and resource intensive. The advent of microplate readers revolutionized bacterial growth studies by enabling high-throughput and real-time monitoring of microbial growth kinetics but their use in anaerobic microbiology has remained limited. Here, we present a workflow for using small-footprint microplate readers and the Growthcurver R package to analyze the kinetic growth metrics of anaerobic bacteria. We benchmarked the small-footprint Cerillo Stratus microplate reader against a BioTek Synergy HTX microplate reader in aerobic conditions using DSM 28618 cultures. The growth rates and carrying capacities obtained from the two readers were statistically indistinguishable. However, the area under the logistic curve was significantly higher in cultures monitored by the Stratus reader. We used the Stratus to quantify the growth responses of anaerobically grown and DSM 29485 to different doses of the toxin sodium arsenite. The growth of and was sensitive to arsenite doses of 1.3 μM and 0.4 μM, respectively. Complete inhibition of growth was achieved at 38 μM arsenite for , and 338 μM in . These results show that the Stratus performs similarly to a leading brand of microplate reader and can be reliably used in anaerobic conditions. We discuss the advantages of the small format microplate readers and our experiences with the Stratus.
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http://dx.doi.org/10.1101/2023.10.10.561742 | DOI Listing |
Virology
January 2025
The Wistar Institute of Anatomy and Biology, Philadelphia, PA, USA. Electronic address:
J-Lat cells are derivatives of the Jurkat CD4 T cell line that contain a non-infectious, inducible HIV provirus with a GFP tag. While these cells have substantially advanced our understanding of HIV latency, their use by many laboratories in low and middle-income countries is restricted by limited access to flow cytometry. To overcome this barrier, we describe a modified J-Lat assay using a standard microplate reader that detects HIV-GFP expression following treatment with latency-reversing agents (LRAs).
View Article and Find Full Text PDFBiol Trace Elem Res
January 2025
Department of Hematology, Affiliated Hospital of Guizhou Medical University, No. 4 Bei Jing Road, Yunyan District, Guiyang, 550004, Guizhou Province, China.
Chronic fluorosis is often accompanied by neurological symptoms, leading to attention, memory and learning ability decline and causing tension, anxiety, depression, and other mental symptoms. In the present study, we analyzed the molecular mechanisms of SIRT1-BDNF regulation of PI3K-AKT, MAPK, and FOXO1A in F-treated BV2 cells. The cytotoxic effect of sodium fluoride (NaF) on BV2 cells was assessed using Cell Counting Kit-8 (CCK-8), crystal violet, and 5-ethynyl-2'-deoxyuridine (EdU) staining.
View Article and Find Full Text PDFTransl Androl Urol
December 2024
Center for Reproductive Medicine, Affiliated Hospital of Nantong University, Nantong, China.
Background: Para-phenylenediamine (PPD) is a crystalline solid that belongs to the aromatic amine group, widely used in the manufacturing of various dyes. PPD exhibits toxic effects on female hormone stability, ovarian function, and embryo development. Although studies have shown that PPD exposure can damage oocyte quality in female mice, research on its effects on male reproductive capability, particularly on human sperm quality and function, is limited.
View Article and Find Full Text PDFBio Protoc
January 2025
Glycometabolic Biochemistry Laboratory, RIKEN Cluster for Pioneering Research, Riken, 2-1 Hirosawa, Wako Saitama, Japan.
Cytosolic peptide:-glycanase (PNGase/NGLY1 in mammals), an amidase classified under EC:3.5.1.
View Article and Find Full Text PDFBMC Cancer
January 2025
School of Life Science and Technology, Shandong Second Medical University, Weifang, Shandong, 261053, P.R. China.
Background: ABCB1 overexpression is a key factor in causing multidrug resistance (MDR). As a result, it is crucial to discover effective medications against ABCB1 to overcome MDR. Falnidamol, a tyrosine kinase inhibitor (TKI) targeting the epidermal growth factor receptor (EGFR), is currently in phase 1 clinical trials for the treatment of solid tumors.
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