PMC

The hasABC genes were expressed in trans under the rofA promoter on pHasABC and transferred to S. pyogenes HSC5 strains whose expression of chromosomal capsule genes was blocked by disrupting the first transcribed capsule gene, hasA. Those complemented strains with pHasABC exhibited the same phenotype of capsule thermoregulation as the strains whose capsule genes were expressed under the native promoter on the chromosome. The following strains were used: CovR⁻ strain (CovR⁻), CovRIFD; chromosomal capsule gene knock-out strain (HasABC⁻), ΩHasA; in trans capsule gene complemented strain (HasABC⁻(pHasABC)), ΩHasA(pHasABC); CovR⁻ HasABC⁻ strain (CovR⁻ HasABC⁻); CovRIFD:ΩHasA; CovR⁻ HasABC⁻ strain complemented with pHasABC (CovR⁻ HasABC⁻ (pHasABC)), CovRIFD:ΩHasA(pHasABC); CovR⁻ CvfA⁻ HasABC⁻ strain (CvfA⁻ CovR⁻ HasABC⁻), CovRIFD:TnCvfA1:ΩHasA; CovR⁻ CvfA⁻ HasABC⁻ strain complemented with pHasABC (CvfA⁻ CovR⁻ HasABC⁻ (pHasABC)), CovRIFD:TnCvfA1:ΩHasA(pHasABC).

A. rim101Δ mutants have a capsule defect. Cells were incubated in capsule inducing conditions for 3 days. Capsule was assessed by staining with India ink and visualizing the zone of exclusion at 63× magnification (scale bar = 10 m). B. The rim101Δ mutant sheds equivalent capsule to wild-type. Electrophoretic mobility and quantity of shed polysaccharide was assessed by a blotting technique of culture medium filtrate, using an anti-GXM antibody to probe for capsule as previously described . Cells were incubated in Dulbecco's modified Eagle's medium or low iron medium for 1 week before filtering. Arrow indicates direction of electrophoresis. C. Gfp-tagged Rim101 is functional. Cells were incubated in capsule inducing conditions for 3 days. Capsule was assessed by staining with India ink and visualizing the zone of exclusion at 63× magnification (scale bar = 10 m). D. Rim101-S773A does not complement capsule. Cells were incubated in capsule inducing conditions for 3 days. Capsule was assessed by staining with India ink and visualizing the zone of exclusion at 63× magnification (scale bar = 10 m).

Comparison of Thy1 in tissue samples taken from normal skin, radiation-naïve capsule (NIR capsule), and irradiated capsule (IR Capsule). A) NIR capsule and IR capsule THY1 mRNA is significantly elevated above skin. B) Elevated mRNA expression translated into increased Thy1 protein, with IR capsule significantly elevated above both NIR capsule and skin. Finally, while NIR capsule Thy1 trended higher than skin, this did not reach significance. * = p<0.05, ** = p<0.01. *** = p<0.001, **** = p<0.0001

Capsule endoscopy publications per type of capsule, 2000 to 2021. Note: Robotic Capsule (RC), Magnetic Capsule (MCE), Colon Capsule (CCE), Esophagus Capsule (ECE), Non-Imaging Capsule (NIC), Small-Bowel Capsule (SBCE).