Integrated optical Mach-Zehnder interferometer as simazine immunoprobe

Immunoassay has become a versatile tool in several fields of analytical chemistry. We describe the characterization and the application of different integrated optical channel waveguide Mach-Zehnder interferometers (MZIs) as label-free immunoprobes. The performance of the classical MZI is compared with that of a modified structure which incorporates a 3x3 coupler. Characterization of the devices demonstrates a dramatic improvement gained by using the 3x3 coupler. Two main advantages are achieved by the modified device. First, the possibility of referencing the output signal allows the elimination of signal fluctuations due to coupling and light-source instabilities. An increase of the signal-to-noise ratio by a factor of up to 10 is achieved. Secondly, the phase shift between the three outputs allows unambiguous detection with optimum sensitivity. For the detection of the herbicide simazine, the functional properties of the transducer surface are optimized by an appropriate chemical modification. Using this improved device, a simazine immunoassay has been carried out with a test midpoint of 0.3 ppb and a detection limit of approximately 0.1 ppb. The excellent performance, established manufacturing techniques and the potential for simplification and parallelization make the device attractive for further development

ILC2-modulated T cell-to-MDSC balance is associated with bladder cancer recurrence.

Non-muscle-invasive bladder cancer (NMIBC) is a highly recurrent tumor despite intravesical immunotherapy instillation with the bacillus Calmette-Guérin (BCG) vaccine. In a prospective longitudinal study, we took advantage of BCG instillations, which increase local immune infiltration, to characterize immune cell populations in the urine of patients with NMIBC as a surrogate for the bladder tumor microenvironment. We observed an infiltration of neutrophils, T cells, monocytic myeloid-derived suppressor cells (M-MDSCs), and group 2 innate lymphoid cells (ILC2). Notably, patients with a T cell-to-MDSC ratio of less than 1 showed dramatically lower recurrence-free survival than did patients with a ratio of greater than 1. Analysis of early and later time points indicated that this patient dichotomy existed prior to BCG treatment. ILC2 frequency was associated with detectable IL-13 in the urine and correlated with the level of recruited M-MDSCs, which highly expressed IL-13 receptor α1. In vitro, ILC2 were increased and potently expressed IL-13 in the presence of BCG or tumor cells. IL-13 induced the preferential recruitment and suppressive function of monocytes. Thus, the T cell-to-MDSC balance, associated with a skewing toward type 2 immunity, may predict bladder tumor recurrence and influence the mortality of patients with muscle-invasive cancer. Moreover, these results underline the ILC2/IL-13 axis as a targetable pathway to curtail the M-MDSC compartment and improve bladder cancer treatment

Storm characteristics influence nitrogen removal in an urban estuarine environment

Sustaining water quality is an important component of coastal resilience. Floodwaters deliver reactive nitrogen (including NOx) to sensitive aquatic systems and can diminish water quality. Coastal habitats in flooded areas can be effective at removing reactive nitrogen through denitrification (DNF). However, less is known about this biogeochemical process in urbanized environments. This study assessed the nitrogen removal capabilities of flooded habitats along an urban estuarine coastline in the upper Neuse River estuary, NC, USA, under two nitrate concentrations (16.8 and 52.3 µM NOx, respectively). We also determined how storm characteristics (e.g., precipitation and wind) affect water column NOx concentrations and consequently DNF by flooded habitats. Continuous flow sediment core incubation experiments quantified gas and nutrient fluxes across the sediment–water interface in marsh, swamp forest, undeveloped open space, stormwater pond, and shallow subtidal sediments. All habitats exhibited net DNF. Additionally, all habitats increased DNF rates under elevated nitrate conditions compared to low nitrate. Structured habitats with high-sediment organic matter had higher nitrogen removal capacity than unstructured, low-sediment organic matter habitats. High-precipitation–high-wind-storm events produced NOx concentrations significantly lower than other types of storms (e.g., low-precipitation–high-wind, high-wind–low-precipitation, low-wind–low-precipitation), which likely results in relatively low DNF rates by flooded habitats and low removal percentages of total dissolved nitrogen loads. These results demonstrate the importance of natural systems to water quality in urbanized coastal areas subject to flooding.</p

Reversible Live-Cell Labeling with Retro-engineered HaloTags Enables Long-Term High- and Super-Resolution Imaging

Self-labeling enzymes (SLE) such as the HaloTag have emerged as powerful tools in high and super-resolution fluorescence microscopy. Newly developed fluorogenic SLE substrates enable imaging in the presence of excess dye. To exploit this feature for reversible labeling, we engineered two variants of HaloTag7 with restored dehalogenase activity. Kinetic studies in vitro showed different turnover kinetics for reHaloTagS (≈0.006 s−1) and reHaloTagF (≈0.055 s−1). Imaging by confocal and stimulated emission depletion microscopy yielded 3-5-time enhanced photostability of reHaloTag labeling. Prominently, single molecule imaging with reHaloTags enabled controlled and stable labeling density over extended time periods. By combination with structured illumination, simultaneous visualization of single molecule diffusion and organellar dynamics was achieved. These applications highlight the potential of reHaloTag labeling for pushing the limits of advanced fluorescence microscopy techniques

Living shorelines can enhance the nursery role of threatened estuarine habitats

Coastal ecosystems provide numerous services, such as nutrient cycling, climate change amelioration, and habitat provision for commercially valuable organisms. Ecosystem functions and processes are modified by human activities locally and globally, with degradation of coastal ecosystems by development and climate change occurring at unprecedented rates. The demand for coastal defense strategies against storms and sea-level rise has increased with human population growth and development along coastlines world-wide, even while that population growth has reduced natural buffering of shorelines. Shoreline hardening, a common coastal defense strategy that includes the use of seawalls and bulkheads (vertical walls constructed of concrete, wood, vinyl, or steel), is resulting in a "coastal squeeze" on estuarine habitats. In contrast to hardening, living shorelines, which range from vegetation plantings to a combination of hard structures and plantings, can be deployed to restore or enhance multiple ecosystem services normally delivered by naturally vegetated shores. Although hundreds of living shoreline projects have been implemented in the United States alone, few studies have evaluated their effectiveness in sustaining or enhancing ecosystem services relative to naturally vegetated shorelines and hardened shorelines. We quantified the effectiveness of (1) sills with landward marsh (a type of living shoreline that combines marsh plantings with an offshore low-profile breakwater), (2) natural salt marsh shorelines (control marshes), and (3) unvegetated bulkheaded shores in providing habitat for fish and crustaceans (nekton). Sills supported higher abundances and species diversity of fishes than unvegetated habitat adjacent to bulkheads, and even control marshes. Sills also supported higher cover of filter-feeding bivalves (a food resource and refuge habitat for nekton) than bulkheads or control marshes. These ecosystem-service enhancements were detected on shores with sills three or more years after construction, but not before. Sills provide added structure and may provide better refuges from predation and greater opportunity to use available food resources for nekton than unvegetated bulkheaded shores or control marshes. Our study shows that unlike shoreline hardening, living shorelines can enhance some ecosystem services provided by marshes, such as provision of nursery habitat