Candida auris Discovery through Community Wastewater Surveillance during Healthcare Outbreak, Nevada, USA, 2022.

Candida auris transmission is steadily increasing across the United States. We report culture-based detection of C. auris in wastewater and the epidemiologic link between isolated strains and southern Nevada, USA, hospitals within the sampled sewershed. Our results illustrate the potential of wastewater surveillance for containing C. auris.

High efficiency Fresnel lens design and fabrication in a two-stage photopolymer.

We show the design and fabrication of high diffraction efficiency, optically recorded gradient-index Fresnel lenses in a two-stage photopolymer. A design analysis reveals that lens f/# is limited by the material refractive index contrast, motivating use of recent high-contrast polymers. The number of pixels required for the optical exposure is typically well beyond available spatial light-modulator resolutions, motivating the use of a photolithographic mask. We use a dithered binary chrome mask with 9000×9000 pixels of 2.5 µm diameter to write lenses up to 23 mm in diameter. Lenses down to f/44 with 76% diffraction efficiency and f/79 with 83% diffraction efficiency are demonstrated.

Controlled Mechanical Property Gradients Within a Digital Light Processing Printed Hydrogel-Composite Osteochondral Scaffold.

Tissue engineered scaffolds are needed to support physiological loads and emulate the micrometer-scale strain gradients within tissues that guide cell mechanobiological responses. We designed and fabricated micro-truss structures to possess spatially varying geometry and controlled stiffness gradients. Using a custom projection microstereolithography (µSLA) system, using digital light projection (DLP), and photopolymerizable poly(ethylene glycol) diacrylate (PEGDA) hydrogel monomers, three designs with feature sizes < 200 µm were formed: (1) uniform structure with 1 MPa structural modulus ( E ) designed to match equilibrium modulus of healthy articular cartilage, (2) E = 1 MPa gradient structure designed to vary strain with depth, and (3) osteochondral bilayer with distinct cartilage ( E = 1 MPa) and bone ( E = 7 MPa) layers. Finite element models (FEM) guided design and predicted the local mechanical environment. Empty trusses and poly(ethylene glycol) norbornene hydrogel-infilled composite trusses were compressed during X-ray microscopy (XRM) imaging to evaluate regional stiffnesses. Our designs achieved target moduli for cartilage and bone while maintaining 68-81% porosity. Combined XRM imaging and compression of empty and hydrogel-infilled micro-truss structures revealed regional stiffnesses that were accurately predicted by FEM. In the infilling hydrogel, FEM demonstrated the stress-shielding effect of reinforcing structures while predicting strain distributions. Composite scaffolds made from stiff µSLA-printed polymers support physiological load levels and enable controlled mechanical property gradients which may improve in vivo outcomes for osteochondral defect tissue regeneration. Advanced 3D imaging and FE analysis provide insights into the local mechanical environment surrounding cells in composite scaffolds.

Microscale Photopatterning of Through-thickness Modulus in a Monolithic and Functionally Graded 3D Printed Part.

3D printing is transforming traditional processing methods for applications ranging from tissue engineering to optics. To fulfill its maximum potential, 3D printing requires a robust technique for producing structures with precise three-dimensional (x, y and z) control of mechanical properties. Previous efforts to realize such spatial control of modulus within 3D printed parts have largely focused on low-resolution (mm to cm scale) multi-material processes and grayscale approaches that spatially vary the modulus in the x-y plane and energy dose-based (E = I 0 t exp) models that do not account for the resin's sub-linear response to irradiation intensity. Here, we demonstrate a novel approach for through-thickness (z) voxelated control of mechanical properties within a single-material, monolithic part. Control over the local modulus is enabled by a predictive model that incorporates the observed non-reciprocal dose response of the material. The model is validated by an application of atomic force microscopy to map the through-thickness modulus on multi-layered 3D parts. Overall, both smooth gradations (30 MPa change over ≈75 µm) and sharp step-changes (30 MPa change over ≈5 µm) in modulus are realized in poly(ethylene glycol) diacrylate based 3D constructs, paving the way for advancements in tissue engineering, stimuli-responsive 4D printing and graded metamaterials.

3D printing of sacrificial thioester elastomers using digital light processing for templating 3D organoid structures in soft biomatrices.

Biofabrication allows for the templating of structural features in materials on cellularly-relevant size scales, enabling the generation of tissue-like structures with controlled form and function. This is particularly relevant for growing organoids, where the application of biochemical and biomechanical stimuli can be used to guide the assembly and differentiation of stem cells and form architectures similar to the parent tissue or organ. Recently, ablative laser-scanning techniques was used to create 3D overhang features in collagen hydrogels at size scales of 10-100µm and supported the crypt-villus architecture in intestinal organoids. As a complementary method, providing advantages for high-throughput patterning, we printed thioester functionalized poly(ethylene glycol) (PEG) elastomers using digital light processing (DLP) and created sacrificial, 3D shapes that could be molded into soft (G' < 1000 Pa) hydrogel substrates. Specifically, three-arm 1.3 kDa PEG thiol and three-arm 1.6 kDa PEG norbornene, containing internal thioester groups, were photopolymerized to yield degradable elastomers. When incubated in a solution of 300 mM 2-mercaptoethanol (pH 9.0), 1 mm thick 10 mm diameter elastomer discs degraded in <2 h. Using DLP, arrays of features with critical dimensions of 37 ± 4µm, resolutions of 22 ± 5µm, and overhang structures as small as 50µm, were printed on the order of minutes. These sacrificial thioester molds with physiologically relevant features were cast-molded into Matrigel and subsequently degraded to create patterned void spaces with high fidelity. Intestinal stem cells (ISCs) cultured on the patterned Matrigel matrices formed confluent monolayers that conformed to the underlying pattern. DLP printed sacrificial thioester elastomer constructs provide a robust and rapid method to fabricate arrays of 3D organoid-sized features in soft tissue culture substrates and should enable investigations into the effect of epithelial geometry and spacing on the growth and differentiation of ISCs.

Detection of early stages of Myxobolus cerebralis in fin clips from rainbow trout (Orynchus mykiss).

A nested polymerase chain reaction (PCR) assay was used to detect early stages of Myxobolus cerebralis in caudal and adipose fin samples from rainbow trout (RT). To determine sensitivity, groups of 10 RT were exposed to 2,000 M. cerebralis triactinomyxons/fish for 1 hour at 15 degrees C and subsequently moved to clean recirculating water. Fish were held for 2 and 6 hours and 1, 2, 3, 5, 7, 10, 30, and 60 days before sampling by nonlethal fin biopsy. Nested PCR performed on fin clips showed that M. cerebralis DNA was detected in caudal fin tissue in 100% of fish up to 5 days postexposure. At days 7 and 10 postexposure, 80% of fish were positive, and at 60 days postexposure, 60% of fish were positive using this technique. Conversely, testing on adipose fin clips proved less sensitive, as positive fish dropped from 80% at day 7 to below 20% at day 10 postinfection. Since detection of M. cerebralis infection using caudal fin samples coupled with nested PCR is an effective method for detection of early parasite stages, use of this technique provides for accurate, nonlethal testing.