Net Ecosystem Carbon Balance in a North Carolina, USA, Salt Marsh

Salt marshes have among the highest carbon (C) burial rates of any ecosystem and often rely on C accumulation to gain elevation and persist in locations with accelerating sea level rise. Net ecosystem carbon balance (NECB), the accumulation or loss of C resulting from vertical CO2 and CH4 gas fluxes, lateral C fluxes, and sediment C inputs, varies across salt marshes; thus, extrapolation of NECB to an entire marsh is challenging. Anthropogenic nitrogen (N) inputs to salt marshes impact NECB by influencing each component of NECB, but differences in the impacts of fertilization between edge and interior marsh must be considered when scaling up. NECB was estimated for the 0.5 km2 Spartina alterniflora marsh area of Freeman Creek, NC, under control and fertilized conditions at both interior and edge berm sites. Annual CO2 fluxes were nearly balanced at control sites, but fertilization significantly increased net CO2 emissions at edge sites. Lateral C export, modeled using respiration rates, represented a significant C loss that increased with fertilization in both edge and interior marsh. Sediment C input was a significant C source in the interior, nearly doubling with fertilization, but represented a small source on the edge. When extrapolating C exchanges to the entire marsh, including edge which comprised 17% of the marsh area, the marsh displayed net loss of C despite a net C gain in the interior. Fertilization increased net C loss fivefold. Extrapolation of NECB to whole marshes requires inclusion of C fluxes for both edge and interior marsh

Reliable solid-state circuits Semiannual report no. 2, Jun. 1 - Nov. 30, 1965

Pulse width modulator and other microminiaturized electronic equipment for space age application

A method for using shoreline morphology to predict suspended sediment concentration in tidal creeks

Improving mechanistic prediction of shoreline response to sea level rise is currently limited by 1) morphologic complexity of tidal creek shorelines that confounds application of mechanistic models, and 2) availability of suspended sediment measurements to parameterize mechanistic models. To address these challenges we developed a metric to distinguish two morphodynamic classes of tidal creek and tested whether this metric could be used to predict suspended sediment concentration. We studied three small tidal creeks in North Carolina, U.S.A. We collected suspended sediment at one non-tidal and two tidal sites in each creek and measured the wetland and channel width using a geographic information system. In each creek, tidal harmonics were measured for one year, sediment accretion on the salt marsh was measured for three years, and shoreline erosion was measured from aerial photographs spanning 50�years. Additional total suspended solids measurements from seven creeks reported in a national database supplemented our analysis. Among the three intensively studied creeks, shoreline erosion was highest in the most embayed creek (having a wider channel than the width of adjoining wetlands) and lowest in the wetland-dominated creek (having a channel narrower than the width of adjoining wetlands). Wetland sediment accretion rate in the wetland-dominated creek was four times higher than the accretion in the embayed creek. The wetland-dominated tidal creek had over twice the suspended sediment as the most embayed creek. Based on these results, we conclude that our metric of embayed and contrasting wetland-dominated creek morphology provides a guide for choosing between two types of morphodynamic models that are widely used to predict wetland shoreline change. This metric also allowed us to parse the 10 tidal creeks studied into two groups with different suspended sediment concentrations. This relationship between suspended sediment concentration and creek morphology provides a method to estimate sediment concentration for individual tidal creek shorelines from spatial data alone, enabling more accurate parameterization of shoreline change models

Ultra-high throughput functional enrichment of large monoamine oxidase (MAO-N) libraries by fluorescence activated cell sorting

Directed evolution enables the improvement and optimisation of enzymes for particular applications and is a valuable tool for biotechnology and synthetic biology. However, studies are often limited in their scope by the inability to screen very large numbers of variants to identify improved enzymes. One class of enzyme for which a universal, operationally simple ultra-high throughput (>106 variants per day) assay is not available is flavin adenine dinucleotide (FAD) dependent oxidases. The current high throughput assay involves a visual, colourimetric, colony-based screen, however this is not suitable for very large libraries and does not enable quantification of the relative fitness of variants. To address this, we describe an optimised method for the sensitive detection of oxidase activity within single Escherichia coli (E. coli) cells, using the monoamine oxidase from Aspergillus niger, MAO-N, as a model system. In contrast to other methods for the screening of oxidase activity in vivo, this method does not require cell surface expression, emulsion formation or the addition of an extracellular peroxidase. Furthermore, we show that fluorescence activated cell sorting (FACS) of large libraries derived from MAO-N under the assay conditions can enrich the library in functional variants at much higher rates than via the colony-based method. We demonstrate its use for directed evolution by identifying a new mutant of MAO-N with improved activity towards a novel secondary amine substrate. This work demonstrates, for the first time, an ultra-high throughput screening methodology widely applicable for the directed evolution of FAD dependent oxidases in E. coli

Liver regulatory mechanisms of noncoding variants at lipid and metabolic trait loci

Genome-wide association studies (GWASs) have identified hundreds of risk loci for liver disease and lipid-related metabolic traits, although identifying their target genes and molecular mechanisms remains challenging. We predicted target genes at GWAS signals by integrating them with molecular quantitative trait loci for liver gene expression (eQTL) and liver chromatin accessibility QTL (caQTL). We predicted specific regulatory caQTL variants at four GWAS signals located near EFHD1, LITAF, ZNF329, and GPR180. Using transcriptional reporter assays, we determined that caQTL variants rs13395911, rs11644920, rs34003091, and rs9556404 exhibit allelic differences in regulatory activity. We also performed a protein binding assay for rs13395911 and found that FOXA2 differentially interacts with the alleles of rs13395911. For variants rs13395911 and rs11644920 in putative enhancer regulatory elements, we used CRISPRi to demonstrate that repression of the enhancers altered the expression of the predicted target and/or nearby genes. Repression of the element at rs13395911 reduced the expression of EFHD1, and repression of the element at rs11644920 reduced the expression of LITAF, SNN, and TXNDC11. Finally, we showed that EFHD1 is a metabolically active gene in HepG2 cells. Together, these results provide key steps to connect genetic variants with cellular mechanisms and help elucidate the causes of liver disease

Young man with anorexia nervosa

non

Methodological Considerations in Scale Refinement with Diverse Populations: A Case Example Using the CES-D with a Community Sample of American Indian Women

Although many psychometric assessments are used extensively in population-based research to determine psychopathology, these tools have not been thoroughly validated or appropriately adapted for use in diverse populations. Indeed, depression measurement studies among American Indian and female populations are scarce, omitting key opportunities to tailor psychological measurement for this population. To build psychometric evidence of measures in this population, we used a procedural method to examine a standard psychological instrument-the Center for Epidemiological Studies Depression Scale (CES-D)-with a community sample of southeast American Indian women. Our results showed strong psychometric reliability of the 20-item CES-D. The "effort" item presented diminished validity, as demonstrated by a negative counter-intuitive item-to-total correlation (ITC) value. Dropping the "effort" item resulted in a 19-item scale with a better fit in the within-group examination of community-based American Indian women. Compared to the 20-item CES-D scale, the revised 19-item measure ("effort" item removed) resulted in minimal changes to women's depression categories. However, we did detect patterns in shifts such that the 19-item scale generally underestimated (i.e., placed women in a lower category) depressive symptoms compared to the 20-item scale. Depending on their study goals, researchers engaging in population-based research should carefully weigh the use of original scales that allow for consistency in reporting with refined scales that fit psychometrically. We present the outlined method as a tool that expands on current approaches in scale refinement, and aids researchers in making more informed decisions regarding refined scales with diverse populations

Sea Level Rise Explains Changing Carbon Accumulation Rates in a Salt Marsh Over the Past Two Millennia

High rates of carbon burial observed in wetland sediments have garnered attention as a potential “natural fix” to reduce the concentration of carbon dioxide (CO2) in Earth's atmosphere. A carbon accumulation rate (CAR) can be determined through various methods that integrate a carbon stock over different time periods, ranging from decades to millennia. Our goal was to assess how CAR changed over the lifespan of a salt marsh. We applied a geochronology to a series of salt marsh cores using both 14C and 210Pb markers to calculate CARs that were integrated between 35 and 2,460 years before present. CAR was 39 g C·m−2·year−1 when integrated over millennia but was upward of 148 g C·m−2·year−1 for the past century. We present additional evidence to account for this variability by linking it to changes in relative sea level rise (RSLR), where higher rates of RSLR were associated with higher CARs. Thus, the CAR calculated for a wetland should integrate timescales that capture the influence of contemporary RSLR. Therefore, caution should be exercised not to utilize a CAR calculated over inappropriately short or long timescales as a current assessment or forecasting tool for the climate change mitigation potential of a wetland