Quantifying the relationship between biofilm reduction and thermal tissue damage on metal implants exposed to alternating magnetic fields.

AIM: Metal implant infections are a devastating problem due to the formation of biofilm which impairs the effectiveness of antibiotics and leads to surgical replacement as definitive treatment. Biofilm on metal implants can be reduced using heat generated by alternating magnetic fields (AMF). In this study, the relationship between implant surface biofilm reduction and surrounding tissue thermal damage during AMF exposure is investigated through numerical simulations. METHODS: Mathematical models of biofilm reduction with heat were created based on in vitro experiments. Simulations were performed to predict the spatial and temporal heating on the implant surface and surrounding tissue when exposed to AMF. RESULTS: The modeling results show that intermittent and slow heating can achieve biofilm reduction with a narrow zone of tissue damage around an implant of less than 3 mm. The results also emphasize that uniformity of implant heating is an extremely important factor impacting the effectiveness of biofilm reduction. For a knee implant, using a target temperature of 75 °C, an intermittent treatment strategy of 15 exposures (10 s to target temperature followed by cooldown) achieved a bacterial CFU reduction of 6-log10 across 25% of the implant surface with less than 3 mm of tissue damage. Alternatively, a single 60 s heating exposure to same temperature achieved a bacterial reduction of 6-log10 across 85% of the implant surface, but with 4 mm of tissue damage. CONCLUSION: Overall, this study demonstrates that with uniform heating to temperatures above 70 °C, an implant surface can be largely reduced of biofilm, with only a few mm of surrounding tissue damage.

A Novel Luciferase Assay For Sensitively Monitoring Myocilin Variants in Cell Culture.

PURPOSE: Primary open angle glaucoma-associated mutations in myocilin (MYOC) cause protein "nonsecretion," rendering secreted MYOC difficult to detect using conventional techniques. This study focused on developing and using an assay that can quickly and easily detect mutant MYOC secretion. METHODS: We fused Gaussia luciferase (eGLuc2) to MYOC variants and expressed the constructs in HEK-293T and NTM-5 cells. Secreted and intracellular levels of MYOC eGLuc2 variants were evaluated by Western blotting and compared to untagged and FLAG-tagged MYOC constructs. Secreted and soluble intracellular MYOC eGLuc2 were measured by a GLuc assay. The secretion of nine additional MYOC mutants was assayed in conditioned media from transfected cells to test the applicability of the assay for monitoring other MYOC variants. RESULTS: Myocilin eGLuc2 behaved similarly to untagged and FLAG-tagged MYOC with respect to secretion, soluble intracellular levels, and in response to drug treatment. The GLuc assay could sensitively detect Y437H MYOC secretion 30 minutes after media change. Gaussia luciferase fused variants followed anticipated trends; nonpathogenic variants (D208E, G244V) were secreted at wild-type (WT) levels, whereas predicted disease-causing variants (C245Y, G246R, E300K, Y437H, I477N) demonstrated substantial secretion defects. Secretion defects caused by the C245Y, G246R, and Y437H mutations were partially rescued by permissive growth temperature. Interestingly, however, this increase in secretion was independent of newly synthesized protein. CONCLUSIONS: Fusion of eGLuc2 to MYOC does not significantly change the behavior of MYOC. This newly developed MYOC reporter system can be used to study engineered MYOC variants and potentially to identify modulators of MYOC secretion and function.

Rayleigh scattering measurements of several fluorocarbon gases.

Integrating nephelometers are commonly used to monitor airborne particulate matter. However, they must be calibrated prior to use. The Rayleigh scattering coefficients (b(RS), Mm(-1)), scattering cross sections (σ(RS), cm(2)), and Rayleigh multipliers for tetrafluoromethane (R-14), sulfur hexafluoride, pentafluoroethane (HFC-125), hexafluoropropene (HFC-216), 1,1,1,2,3,3,3,-heptafluoropropane (HFC-227ea), and octafluorocyclobutane (C-318) are reported from measurements made using a Radiance Research M903 integrating nephelometer operating at λ = 530 nm and calibration with gases of known scattering constants. Rayleigh multipliers (±90% conf. int.) were found to be 2.6 ± 0.5, 6.60 ± 0.07, 7.5 ± 1, 14.8 ± 0.9, 15.6 ± 0.5, and 22.3 ± 0.8 times that of air, respectively. To the best of our knowledge, these are the first reported values for R-14, HFC-216, HFC-125, and C-318. Experimental accuracy is supported through measurements of values for SF(6) and HFC-227ea which agree to within 3% of previous literature reports. In addition to documenting fundamental Rayleigh scattering data for the first time, the information presented within will find use for calibration of optical scattering sensors such as integrating nephelometers.