The Staphylococcus aureus cidABC and lrgAB operons have been shown to regulate murein hydrolase activity and affect antibiotic tolerance. The cid operon enhances murein hydrolase activity and antibiotic sensitivity, whereas the lrg operon inhibits these processes. Based on these findings and the structural similarities of the cidA and lrgA gene products to the bacteriophage holin family of proteins, we have proposed that the cid and lrg operons encode holin-and antiholin-like proteins, respectively, that function to control the murein hydrolase activity produced by the bacteria. Analysis of cid operon transcription revealed the presence of two transcripts, one spanning all three cid genes and whose expression is induced by growth in the presence of acetic acid and the other spanning cidB and cidC only that is produced in a sigma B-dependent manner. The cidABC operon lies immediately downstream from the cidR gene, encoding a potential LysR-type transcriptional regulator. In this study, we demonstrate that cidR is involved in the regulation of cidABC expression. Northern blot analyses revealed that the cidR gene product positively regulates cidABC expression by increasing transcription in the presence of acetic acid produced as a result of the metabolism of glucose. As expected for an operon that encodes a positive effector of murein hydrolase activity, the upregulation of cidABC expression resulted in increased murein hydrolase activity produced by these cells. Furthermore, it was demonstrated that antibiotic tolerance and stationary-phase survival of S. aureus are affected by the cidR gene. Taken together, these results demonstrate that the cidR gene product functions as a transcriptional activator of cidABC transcription in response to acetic acid accumulation in the growth medium.

See next page for additional authorsThis is an author-produced, peer-reviewed version of this article. © 2009, Elsevier. Licensed under the Creative Commons AttributionNonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/). The final, definitive version of this document can be found online at Neurobiology of Disease, doi: 10.1016Disease, doi: 10. /j.nbd.2010

The mechanical behavior of soft connective tissue is governed by a dense network of fibrillar proteins in the extracellular matrix. Characterization of this fibrous network requires the accurate extraction of descriptive structural parameters from imaging data, including fiber dispersion and mean fiber orientation. Common methods to quantify fiber parameters include fast Fourier transforms (FFT) and structure tensors, however, information is limited on the accuracy of these methods. In this study, we compared these two methods using test images of fiber networks with varying topology. The FFT method with a band-pass filter was the most accurate, with an error of 0.71 ± 0.43 degrees in measuring mean fiber orientation and an error of 7.4 ± 3.0% in measuring fiber dispersion in the test images. The accuracy of the structure tensor method was approximately 4 times worse than the FFT band-pass method when measuring fiber dispersion. A free software application, FiberFit, was then developed that utilizes an FFT band-pass filter to fit fiber orientations to a semicircular von Mises distribution. FiberFit was used to measure collagen fibril organization in confocal images of bovine ligament at magnifications of 63x and 20x. Grayscale conversion prior to FFT analysis gave the most accurate results, with errors of 3.3 ± 3.1 degrees for mean fiber orientation and 13.3 ± 8.2% for fiber dispersion when measuring confocal images at 63x. By developing and validating a software application that facilitates the automated analysis of fiber organization, this study can help advance a mechanistic understanding of collagen networks and help clarify the mechanobiology of soft tissue remodeling and repair.

This study demonstrates the growth and differentiation of C2C12 myoblasts into functional myotubes on 3-dimensional graphene foam bioscaffolds. Specifically, we establish both bare and laminin coated graphene foam as a biocompatible platform for muscle cells and identify that electrical coupling stimulates cell activity. Cell differentiation and functionality is determined by the expression of myotube heavy chain protein and Ca2+ fluorescence, respectively. Further, our data show that the application of a pulsed electrical stimulus to the graphene foam initiates myotube contraction and subsequent localized substrate movement of over 100 micrometers. These findings will further the development of advanced 3-dimensional graphene platforms for therapeutic applications and tissue engineering.