Selective Optical Imaging for Detection of Bacterial Biofilms in Tissues.

SIGNIFICANCE: The development of an imaging technique to accurately identify biofilm regions on tissues and in wounds is crucial for the implementation of precise surface-based treatments, leading to better patient outcomes and reduced chances of infection. AIM: The goal of this study was to develop an imaging technique that relies on selective trypan blue (TB) staining of dead cells, necrotic tissues, and bacterial biofilms, to identify biofilm regions on tissues and wounds. APPROACH: The study explored combinations of ambient multi-colored LED lights to obtain maximum differentiation between stained biofilm regions and the underlying chicken tissue or glass substrate during image acquisition. The TB imaging results were then visually and statistically compared to fluorescence images using a shape similarity measure. RESULTS: The comparisons between the proposed TB staining method and the fluorescence standard used to detect biofilms on tissues and glass substrates showed up to 97 percent similarity, suggesting that the TB staining method is a promising technique for identifying biofilm regions. CONCLUSIONS: The TB staining method demonstrates significant potential as an effective imaging technique for the identification of fluorescing and non-fluorescing biofilms on tissues and in wounds. This approach could lead to improved precision in surface-based treatments and better patient outcomes.

Cold Atmospheric Pressure Plasma Device Exhibits Etching Effects on Bacterial Biofilms.

Cold atmospheric pressure plasma (CAP) treatment has been shown to kill bacteria and remove bacterial biofilms from surfaces. Here we report the etch capacity of a linear discharge CAP device on Pseudomonas fluorescens biofilms. A 21 kHz, 1.4 kV RMS AC voltage applied to the CAP electrodes generated a hydrated Ar plasma between the plates, with the gas flow directing the plasma species toward the biological sample, causing both bacterial killing and etching of the biofilm. Typical discharge currents for a 2.4 cm long, 0.6 mm wide linear discharge device were 1-4.4 mA. Hydrated Ar flow gas was critical for removal of biofilm from a stainless steel substrate, while both hydrated and dry Ar + O2, Ar + air, O2 only, and air only flow gas mixtures did not cause etching at equivalent or greater discharge current intensities. A biofilm etch rate of > 2 µm/min was achieved, provided the plasma discharge was within 1-2 mm of the substrate surface and used a hydrated Ar gas flow of at least 5 LPM.

Fabrication and Performance of a Multi-Discharge Cold Atmospheric Pressure Plasma Array.

Cold atmospheric pressure plasma (CAP) has been shown to kill bacteria and remove biofilms. Here we report the development of a unique CAP array device consisting of a parallel stack of eight linear-discharge plasma elements that create a ~ 5 cm2 (2.4 cm × 2 cm) treatment area. The CAP device is fabricated from Low Temperature Co-fired Ceramic (LTCC) layers to create 24 mm long linear-discharge channels (500 µm gap) with embedded opposing silver metal electrodes. A 20 kHz AC voltage (0.5-5 kV) applied to the electrodes generates an Ar/O2 plasma between the plates, with the gas flow directing the reactive species toward the biological sample (biofilms, etc.) to affect the antimicrobial treatment. External ballast resistors were used to study discharge uniformity in the stacked array elements and internal thick film ballast resistors (≈150 kΩ) were developed to create a fully integrated device. Typical element discharge currents were 1-2.5 mA with the total array current tested at 20 mA to provide optimal device uniformity. The plasma discharge was further shown to produce reactive hydrogen peroxide and exert antimicrobial effects on Pseudomonas biofilms and Salmonella contaminated eggshell samples, with >99% of the bacterial cells killed with less than 60 seconds of plasma exposure.

Center of Biomedical Research Excellence in Matrix Biology: Building Research Infrastructure, Supporting Young Researchers, and Fostering Collaboration.

The Center of Biomedical Research Excellence in Matrix Biology strives to improve our understanding of extracellular matrix at molecular, cellular, tissue, and organismal levels to generate new knowledge about pathophysiology, normal development, and regenerative medicine. The primary goals of the Center are to i) support junior investigators, ii) enhance the productivity of established scientists, iii) facilitate collaboration between both junior and established researchers, and iv) build biomedical research infrastructure that will support research relevant to cell-matrix interactions in disease progression, tissue repair and regeneration, and v) provide access to instrumentation and technical support. A Pilot Project program provides funding to investigators who propose applying their expertise to matrix biology questions. Support from the National Institute of General Medical Sciences at the National Institutes of Health that established the Center of Biomedical Research Excellence in Matrix Biology has significantly enhanced the infrastructure and the capabilities of researchers at Boise State University, leading to new approaches that address disease diagnosis, prevention, and treatment. New multidisciplinary collaborations have been formed with investigators who may not have previously considered how their biomedical research programs addressed fundamental and applied questions involving the extracellular matrix. Collaborations with the broader matrix biology community are encouraged.

A Cold Atmospheric Pressure Plasma Discharge Device Exerts Antimicrobial Effects.

A cold atmospheric pressure plasma device was developed using two parallel plates of Low Temperature Co-fired Ceramic with embedded electrodes. The 2.4 cm wide by 1 mm deep plasma discharge operates at 20 kHz with a 2-5 kV AC drive signal across a 0.25 mm gap. Mixed Argon/oxygen plasmas were directed between the plates to flow toward a bacterial biofilm sample for treatment. Results showed that at 4-5 kV the plasma etched away a bacterial biofilm on glass in 10 minutes. In addition, we showed that short plasma treatments rapidly killed biofilm resident bacteria with ED90 values of <15 s.

Electromagnetic waves' effect on airflow during air sparging.

Air sparging is a popular, yet slow, remediation technology for soil and groundwater contaminated with volatile organic compounds (VOCs). This paper theoretically and experimentally studies the effect of electromagnetic (EM) waves on air-channel formation within a glass-bead medium-used as an analogy to soil-during air-sparging experiments. The impact of EM waves on cleanup is not the focus of this paper, and the impact on airflow may or may not positively impact resulting cleanup process using air sparging to remove VOCs through volatilization. The hypothesis is that dielectrophoretic forces by EM waves can be used to alter airflow. Air injection was performed at different pressures, in the presence of EM waves (referred to as EM-stimulated) of various power and frequencies and the absence of EM waves (referred to as unstimulated). Digital images of the airflow patterns were collected, processed, and analyzed for all tests. The shape of the zone of influence (ZOI) was observed, and the radius of the zone of influence (ROI) was measured, which showed a 16% increase in ROI due to EM stimulation. An experimentally validated numerical simulation of the electric-field component of EM waves was developed. The correlation between EM-wave and air sparging characteristics were then studied using the numerical simulation and acquired digital images of the airflow to investigate and validate that the dielectrophoretic mechanism is behind the EM effect on airflow.

Aggressive prostate cancer is prevented in ERαKO mice and stimulated in ERßKO TRAMP mice.

Previous evidence suggests soy genistein may be protective against prostate cancer, but whether this protection involves an estrogen receptor (ER)-dependent mechanism is unknown. To test the hypothesis that phytoestrogens may act through ERα or ERß to play a protective role against prostate cancer, we bred transgenic mice lacking functional ERα or ERß with transgenic adenocarcinoma of mouse prostate (TRAMP) mice. Dietary genistein reduced the incidence of cancer in ER wild-type (WT)/transgenic adenocarcinoma of mouse prostate mice but not in ERα knockout (KO) or ERßKO mice. Cancer incidence was 70% in ERWT mice fed the control diet compared with 47% in ERWT mice fed low-dose genistein (300 mg/kg) and 32% on the high-dose genistein (750 mg/kg). Surprisingly, genistein only affected the well differentiated carcinoma (WDC) incidence but had no effect on poorly differentiated carcinoma (PDC). No dietary effects have been observed in either of the ERKO animals. We observed a very strong genotypic influence on PDC incidence, a protective effect in ERαKO (only 5% developed PDC), compared with 19% in the ERWT, and an increase in the incidence of PDC in ERßKO mice to 41%. Interestingly, immunohistochemical analysis showed ERα expression changing from nonnuclear in WDC to nuclear in PDC, with little change in ERß location or expression. In conclusion, genistein is able to inhibit WDC in the presence of both ERs, but the effect of estrogen signaling on PDC is dominant over any dietary treatment, suggesting that improved differential targeting of ERα vs. ERß would result in prevention of advanced prostate cancer.

PS-b-P3HT copolymers as P3HT/PCBM interfacial compatibilizers for high efficiency photovoltaics.

A conducting diblock copolymer of PS-b-P3HT was added to serve as a compatibilizer in a P3HT/PCBM blend, which improved the power-conversion efficiency from 3.3% to 4.1% due to the enhanced crystallinity, morphology, interface interaction, and depth profile of PCBM.

Depletion at solid/liquid interfaces: flowing hexadecane on functionalized surfaces.

We present a neutron reflectivity study on interfaces in contact with flowing hexadecane, which is known to exhibit surface slip on functionalized solid surfaces. The single crystalline silicon substrates were either chemically cleaned Si(100) or Si(100) coated by octadecyl-trichlorosilane (OTS), which resulted in different interfacial energies. The liquid was sheared in situ and changes in reflectivity profiles were compared to the static case. For the OTS surface, the temperature dependence was also recorded. For both types of interfaces, density depletion of the liquid at the interface was observed. In the case of the bare Si substrate, shear load altered the structure of the depletion layer, whereas for the OTS covered surface no effect of shear was observed. Possible links between the depletion layer and surface slip are discussed. The results show that, in contrast to water, for hexadecane the enhancement of the depletion layer with temperature and interfacial energy reduces the amount of slip. Thus a density depletion cannot be the origin of surface slip in this system.

Zinc deficiency increases plasma lipids and atherosclerotic markers in LDL-receptor-deficient mice.

Low zinc concentration can be associated with an increased risk of cardiovascular diseases. In the current study, we hypothesize that zinc deficiency can increase and zinc supplementation can decrease proatherosclerotic events in LDL receptor knock-out (LDL-R-/-) mice fed a moderate-fat diet. Mice were fed either a zinc-deficient (0 micromol Zn/g), a control (0.45 micromol Zn/g), or a zinc-supplemented (1.529 micromol Zn/g) diet for 4 wk. Mice fed the zinc-deficient diet had significantly increased concentrations of cholesterol and triacylglycerides in the VLDL and HDL fractions. Zinc supplementation decreased these lipid variables compared with control mice. We detected significantly higher concentrations of glutathione reductase mRNA in the thoracic aortae of zinc-deficient mice. Furthermore, inflammatory markers, such as nuclear factor-kappaB and vascular cell adhesion molecule-1, were significantly increased in zinc-deficient mice compared with mice of the control or supplemented groups. In addition, zinc deficiency significantly reduced the DNA binding activity of peroxisome proliferator activate receptors (PPARs) in liver extracts. Interestingly, mRNA expression levels of PPARgamma were significantly increased in thoracic aortae of zinc-deficient mice, indicating an adaptation process to decreased PPAR signaling. These data provide in vivo evidence of zinc deficiency inducing proinflammatory events in an atherogenic mouse model. These data also suggest that adequate zinc may be a critical component in protective PPAR signaling during atherosclerosis.