Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Evidence for natural recombination between mink enteritis virus and canine parvovirus

Abstract A virus was isolated from mink showing clinical and pathological signs of enteritis in China. This virus, designated MEV/LN-10, was identified as mink enteritis virus (MEV) based on its cytopathic effect in the feline F81 cell line, the hemagglutination (HA) and hemagglutination inhibition (HI) assay, electron microscopy (EM) and animal infection experiments. The complete viral genome was cloned and sequenced. Phylogenetic and recombination analyses on the complete MEV/LN-10 genome showed evidence of recombination between MEV and canine parvovirus (CPV). The genome was composed of the NS1 gene originating from CPV while the VP1 gene was of MEV origin. This is the first demonstration of recombination between a CPV and MEV in nature. Our findings not only provide valuable evidence indicating that recombination is an important genetic mechanism contributing to the variation and evolution of MEV, but also that heterogeneous recombination can occur in the feline parvovirus subspecies.</p

Evaluation of the Glaucomatous Macular Damage by Chromatic Pupillometry

Abstract Introduction This study aimed to examine the performance of binocular chromatic pupillometry for the objective and rapid detection of primary open-angle glaucoma (POAG), and to explore the association between pupillary light response (PLR) features and structural glaucomatous macular damage. Methods Forty-six patients (mean age 41.00 ± 13.03 years) with POAG and 23 healthy controls (mean age 42.00 ± 11.08 years) were enrolled. All participants underwent sequenced PLR tests of full-field, superior/inferior quadrant-field chromatic stimuli using a binocular head-mounted pupillometer. The constricting amplitude, velocity, and time to max constriction/dilation, and the post-illumination pupil response (PIPR) were analyzed. The inner retina thickness and volume measurements were determined by spectral domain optical coherence tomography. Results In the full-field stimulus experiment, time to pupil dilation was inversely correlated with perifoveal thickness (r = − 0.429, P < 0.001) and perifoveal volume (r = − 0.364, P < 0.001). Dilation time (AUC 0.833) showed good diagnostic performance, followed by the constriction amplitude (AUC 0.681) and PIPR (AUC 0.620). In the superior quadrant-field stimulus experiment, time of pupil dilation negatively correlated with inferior perifoveal thickness (r = − 0.451, P < 0.001) and inferior perifoveal volume (r = − 0.417, P < 0.001). The dilation time in response to the superior quadrant-field stimulus showed the best diagnostic performance (AUC 0.909). In the inferior quadrant-field stimulus experiment, time to pupil dilation (P < 0.001) correlated well with superior perifoveal thickness (r = − 0.299, P < 0.001) and superior perifoveal volume (r = − 0.304, P < 0.001). Conclusion The use of chromatic pupillometry offers a patient-friendly and objective approach to detect POAG, while the impairment of PLR features may serve as a potential indicator of structural macular damage