Kenneth A. Cornell

The study evaluates the efficacy of an image-guided CAP treatment method with a plasma device capable of rapid biofilm removal from chicken tissue. The plasma treatment operating configuration includes a gas mixture of Argon and H₂O at a flowrate of 1.5 lpm. An X-Y stage was used to move the chicken sample below the stationary plasma scalpel at a speed of 0.1 mm/s. The discharge voltage and current were maintained between 3.2 and 3.7 kV (AC 20 kHz), and at 3 mA, respectively. The electrode gap and sample distance were set to 0.6 mm and 4 mm. This configuration facilitated effective biofilm removal, as confirmed by CFU analysis and 3D microscopic analysis showing a >99% reduction in biofilm post treatment with an etch rate of 2.2–5.8 µm/s and an impact width of up to 300 µm. The plasma scalpel electrode temperature reached 94.7 °C, while the targeted biofilm area was heated to 36.3 °C, suggesting non-thermal biofilm disruption. Three-dimensional microscopic analysis revealed biofilm thickness on chicken tissues ranging from 20 to 180 µm, comparable to biofilm loads on mammalian tissues. In conclusion, the study highlights the potential of CAP devices as a promising solution for biofilm debridement.

This paper describes the principles behind the radio-frequency (RF) sensing of bacterial biofilms in pipes and heat exchangers in a dairy processing plant using an electromagnetic simulation. Biofilm formation in dairy processing plants is a common issue where the absence of timely detection and subsequent cleaning can cause serious illness. Biofilms are known for causing health issues and cleaning requires a large volume of water and harsh chemicals. In this work, milk transportation pipes are considered circular waveguides, and pasteurizers/heat exchangers are considered resonant cavities. Simulations were carried out using the CST studio suite high-frequency solver to determine the effectiveness of the real-time RF sensing. The respective dielectric constants and loss tangents were applied to milk and biofilm. In our simulation, it was observed that a 1 µm thick layer of biofilm in a milk-filled pipe shifted the reflection coefficient of a 10.16 cm diameter stainless steel circular waveguide from 0.229 GHz to 0.19 GHz. Further sensitivity analysis revealed a shift in frequency from 0.8 GHz to 1.2 GHz for a film thickness of 5 µm to 10 µm with the highest wave reflection (S11) peak of ≈−120 dB for a 6 µm thick biofilm. A dielectric patch antenna to launch the waves into the waveguide through a dielectric window was also designed and simulated. Simulation using the antenna demonstrated a similar S11 response, where a shift in reflection coefficient from 0.229 GHz to 0.19 GHz was observed for a 1 µm thick biofilm. For the case of the resonant cavity, the same antenna approach was used to excite the modes in a 0.751 m × 0.321 m × 170 m rectangular cavity with heat exchange fins and filled with milk and biofilm. The simulated resonance frequency shifted from 1.52 GHz to 1.54 GHz, for a film thickness varying from 1 µm to 10 µm. This result demonstrated the sensitivity of the microwave detection method. Overall, these results suggest that microwave sensing has promise in the rapid, non-invasive, and real-time detection of biofilm formation in dairy processing plants.

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 (CAP) planar and radial arrays have been developed to both inactivate and remove biofilms from surfaces [1] , [2] . The planar arrays are made from Low Temperature Co-fired Ceramic (LTCC), consisting of 8 linear discharge elements 2.4 cm long with a 0.75 mm gap between each element, and operate at 1-3 kV, 20 kHz AC. Typical gas flows use Ar at 13 SLM hydrated with water. Previous results show generation of hydrogen peroxide and hydroxyl radicals which increase linearly with plasma exposure times from the arrays and >99% Colony Forming Unit (CFU) reduction in <60 s for bacterial biofilms on a range of substrates (stainless steel, glass, plastic) [1] . Here we apply these arrays on a much larger surface area (10× 10 cm) by scanning the active array across a surface. The CFU count, chemical assays, and etch rates for this experiment will be presented. The radial arrays consist of four LTCC elements ≈3 cm in diameter with ≈0.75 cm gaps between each element. These radial arrays are designed for use in pipes. Imaging of biofilm removal is also under development.

The first line therapy for Lyme disease is treatment with doxycycline, amoxicillin, or cefuroxime. In endemic regions, the persistence of symptoms in many patients after completion of antibiotic treatment remains a major healthcare concern. The causative agent of Lyme disease is a spirochete, Borrelia burgdorferi, an extreme auxotroph that cannot exist under free-living conditions and depends upon the tick vector and mammalian hosts to fulfill its nutritional needs. Despite lacking all major biosynthetic pathways, B. burgdorferi uniquely possesses three homologous and functional methylthioadenosine/S-adenosylhomocysteine nucleosidases (MTANs: Bgp, MtnN, and Pfs) involved in methionine and purine salvage, underscoring the critical role these enzymes play in the life cycle of the spirochete. At least one MTAN, Bgp, is exceptional in its presence on the surface of Lyme spirochetes and its dual functionality in nutrient salvage and glycosaminoglycan binding involved in host-cell adherence. Thus, MTANs offer highly promising targets for discovery of new antimicrobials. Here we report on our studies to evaluate five nucleoside analogs for MTAN inhibitory activity, and cytotoxic or cytostatic effects on a bioluminescently engineered strain of B. burgdorferi. All five compounds were either alternate substrates and/or inhibitors of MTAN activity, and reduced B. burgdorferi growth. Two inhibitors: 5′-deoxy-5′-iodoadenosine (IADO) and 5′-deoxy-5′-ethyl-immucillin A (dEt-ImmA) showed bactericidal activity. Thus, these inhibitors exhibit high promise and form the foundation for development of novel and effective antimicrobials to treat Lyme disease.

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.

West Nile virus (WNV) causes potentially fatal neuroinvasive disease and persists at endemic levels in many parts of the world. Despite advances in our understanding of WNV pathogenesis, there remains a significant need for a human vaccine. The domain III (DIII) region of the WNV envelope protein contains epitopes that are the target of neutralizing antibodies. We have constructed a chimeric fusion of the non-toxic cholera toxin (CT) CTA₂/B domains to DIII for investigation as a novel mucosally-delivered WNV vaccine. Purification and assembly of the chimera, as well as receptor-binding and antigen delivery, were verified by western blot, GM1 ELISA and confocal microscopy. Groups of BALB/c mice were immunized intranasally with DIII-CTA₂/B, DIII, DIII mixed with CTA₂/B, or CTA₂/B control, and boosted at 10 days. Analysis of serum IgG after 14 and 45 days revealed that mucosal immunization with DIII-CTA₂/B induced significant DIII-specific humoral immunity and drove isotype switching to IgG2a. The DIII-CTA₂/B chimera also induced antigen-specific IgM and IgA responses. Bactericidal assays indicate that the DIII-CTA₂/B immunized mice produced DIII-specific antibodies that can trigger complement-mediated killing. A dose escalation resulted in increased DIII-specific serum IgG titers on day 45. DIII antigen alone, in the absence of adjuvant, also induced significant systemic responses after intranasal delivery. Our results indicate that the DIII-CTA₂/B chimera is immunogenic after intranasal delivery and merits further investigation as a novel WNV vaccine candidate.

A recombinant form of Escherichia coli 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (E.C. 3.2.2.9) has been purified to homogeneity and crystallized using the hanging-drop vapour-diffusion technique. While several different crystallization conditions were obtained, only one set of conditions yielded crystals suitable for X-ray diffraction analysis. These crystals grow as diamond-shaped wedges, with unit-cell parameters a = 50.92, b = 133.99, c = 70.88 A, alpha = beta = gamma = 90 degrees. The crystals belong to space group P2(1)2(1)2 and diffract to a minimum d spacing of 2.3 A on a MAR345 image plate with a Rigaku RU-200 rotating-anode X-ray generator. On the basis of density calculations, two monomers are predicted per asymmetric unit (Matthews coefficient, V(M) = 2.37 A(3) Da(-1)), with a solvent content of 48%.