The ‘Hestia Project’ uses a bottom-up approach to quantify fossil fuel CO₂(FFCO₂) emissions spatially at the building/street level and temporally at the hourly level. Hestia FFCO₂ emissions are provided in the form of a group of sector-specific vector layers with point, line, and polygon sources to support carbon cycle science and climate policy. Application to carbon cycle science, in particular, requires regular gridded data in order to link surface carbon fluxes to atmospheric transport models. However, the heterogeneity and complexity of FFCO₂ sources within regular grids is sensitive to spatial resolution. From the perspective of a data provider, we need to find a balance between resolution and data volume so that the gridded data product retains the maximum amount of information content while maintaining an efficient data volume.

The Shannon entropy determines the minimum bits that are needed to encode an information source and can serve as a metric for the effective information content. In this paper, we present an analysis of the Shannon entropy of gridded FFCO₂ emissions with varying resolutions in four Hestia study areas, and find: (1) the Shannon entropy increases with smaller grid resolution until it reaches a maximum value (the max-entropy resolution); (2) total emissions (the sum of several sector-specific emission fields) show a finer max-entropy resolution than each of the sector-specific fields; (3) the residential emissions show a finer max-entropy resolution than the commercial emissions; (4) the max-entropy resolution of the onroad emissions grid is closely correlated to the density of the road network. These findings suggest that the Shannon entropy can detect the information effectiveness of the spatial resolution of gridded FFCO₂ emissions. Hence, the resolution-entropy relationship can be used to assist in determining an appropriate spatial resolution for urban CO₂ flux studies. We conclude that the optimal spatial resolution for providing Hestia total FFCO₂ emissions products is centered around 100 m, at which the FFCO₂ emissions data can not only fully meet the requirement of urban flux integration, but also be effectively used in understanding the relationships between FFCO₂ emissions and various social-economic variables at the U.S. census block group level.

Atmospheric CO₂ inversions estimate surface carbon fluxes from an optimal fit to atmospheric CO₂ measurements, usually including prior constraints on the flux estimates. Eleven sets of carbon flux estimates are compared, generated by different inversions systems that vary in their inversions methods, choice of atmospheric data, transport model and prior information. The inversions were run for at least 5 yr in the period between 1990 and 2010. Mean fluxes for 2001–2004, seasonal cycles, interannual variability and trends are compared for the tropics and northern and southern extra-tropics, and separately for land and ocean. Some continental/basin-scale subdivisions are also considered where the atmospheric network is denser. Four-year mean fluxes are reasonably consistent across inversions at global/latitudinal scale, with a large total (land plus ocean) carbon uptake in the north (−3.4 Pg C yr^-1 (±0.5 Pg C yr^-1 standard deviation), with slightly more uptake over land than over ocean), a significant although more variable source over the tropics (1.6 ± 0.9 Pg C yr^-1) and a compensatory sink of similar magnitude in the south (−1.4 ± 0.5 Pg C yr^-1) corresponding mainly to an ocean sink. Largest differences across inversions occur in the balance between tropical land sources and southern land sinks. Interannual variability (IAV) in carbon fluxes is larger for land than ocean regions (standard deviation around 1.06 versus 0.33 Pg C yr[superscript −1] for the 1996–2007 period), with much higher consistency among the inversions for the land. While the tropical land explains most of the IAV (standard deviation ~ 0.65 Pg C yr^-1), the northern and southern land also contribute (standard deviation ~ 0.39 Pg C yr^-1). Most inversions tend to indicate an increase of the northern land carbon uptake from late 1990s to 2008 (around 0.1 Pg C yr^-1, predominantly in North Asia. The mean seasonal cycle appears to be well constrained by the atmospheric data over the northern land (at the continental scale), but still highly dependent on the prior flux seasonality over the ocean. Finally we provide recommendations to interpret the regional fluxes, along with the uncertainty estimates.

Quantifying greenhouse gas (GHG) emissions from cities is a key challenge towards effective emissions management. An inversion analysis from the INdianapolis FLUX experiment (INFLUX) project, as the first of its kind, has achieved a top-down emission estimate for a single city using CO₂ data collected by the dense tower network deployed across the city. However, city-level emission data, used as a priori emissions, are also a key component in the atmospheric inversion framework. Currently, fine-grained emission inventories (EIs) able to resolve GHG city emissions at high spatial resolution, are only available for few major cities across the globe. Following the INFLUX inversion case with a global 1.1 km ODIAC fossil fuel CO₂ emission dataset, we further improved the ODIAC emission field and examined its utility as a prior for the city scale inversion. We disaggregated the 1.1 km ODIAC non-point source emissions using geospatial datasets such as the global road network data and satellite-data driven surface imperviousness data to a 30 . 30 m resolution. We assessed the impact of the improved emission field on the inversion result, relative to priors in previous studies (Hestia and ODIAC). The posterior total emission estimate (5.1 MtC/yr) remains statistically similar to the previous estimate with ODIAC (5.3 MtC/yr). However, the distribution of the flux corrections was very close to those of Hestia inversion and the model-observation mismatches were significantly reduced both in forward and inverse runs, even without hourly temporal changes in emissions. EIs reported by cities often do not have estimates of spatial extents. Thus, emission disaggregation is a required step when verifying those reported emissions using atmospheric models. Our approach offers gridded emission estimates for global cities that could serves as a prior for inversion, even without locally reported EIs in a systematic way to support city-level Measuring, Reporting and Verification (MRV) practice implementation.

Low fluid shear force, including that encountered in microgravity models, induces bacterial responses, but the range of bacteria capable of responding to this signal remains poorly characterized. We systematically analyzed a range of Gram negative Enterobacteriaceae for conservation of the low-shear modeled microgravity (LSMMG) response using phenotypic assays, qPCR, and targeted mutations. Our results indicate LSMMG response conservation across Enterobacteriacae with potential variance in up- or down-regulation of a given response depending on genus. Based on the data, we analyzed the role of the trp operon genes and the TrpR regulator in the LSMMG response using targeted mutations in these genes in S. Typhimurium and E. coli. We found no alteration of the LSMMG response compared to WT in these mutant strains under the conditions tested here. To our knowledge, this study is first-of-kind for Citrobacter, Enterobacter, and Serratia, presents novel data for Escherichia, and provides the first analysis of trp genes in LSMMG responses. This impacts our understanding of how LSMMG affects bacteria and our ability to modify bacteria with this condition in the future.

This paper presents an analysis of methane emissions from the Los Angeles Basin at monthly timescales across a 4-year time period – from September 2011 to August 2015. Using observations acquired by a ground-based near-infrared remote sensing instrument on Mount Wilson, California, combined with atmospheric CH₄–CO₂ tracer–tracer correlations, we observed −18 to +22 % monthly variability in CH₄ : CO₂ from the annual mean in the Los Angeles Basin. Top-down estimates of methane emissions for the basin also exhibit significant monthly variability (−19 to +31 % from annual mean and a maximum month-to-month change of 47 %). During this period, methane emissions consistently peaked in the late summer/early fall and winter. The estimated annual methane emissions did not show a statistically significant trend over the 2011 to 2015 time period.

A distinct pathovar of Salmonella enterica serovar Typhimurium, ST313, has emerged in sub-Saharan Africa as a major cause of fatal bacteremia in young children and HIV-infected adults. D23580, a multidrug resistant clinical isolate of ST313, was previously shown to have undergone genome reduction in a manner that resembles that of the more human-restricted pathogen, Salmonella enterica serovar Typhi. It has since been shown through tissue distribution studies that D23580 is able to establish an invasive infection in chickens. However, it remains unclear whether ST313 can cause lethal disease in a non-human host following a natural course of infection. Herein we report that D23580 causes lethal and invasive disease in a murine model of infection following peroral challenge. The LD⁵⁰ of D23580 in female BALB/c mice was 4.7 x 10⁵ CFU. Tissue distribution studies performed 3 and 5 days post-infection confirmed that D23580 was able to more rapidly colonize the spleen, mesenteric lymph nodes and gall bladder in mice when compared to the well-characterized S. Typhimurium strain SL1344. D23580 exhibited enhanced resistance to acid stress relative to SL1344, which may lend towards increased capability to survive passage through the gastrointestinal tract as well as during its intracellular lifecycle. Interestingly, D23580 also displayed higher swimming motility relative to SL1344, S. Typhi strain Ty2, and the ST313 strain A130. Biochemical tests revealed that D23580 shares many similar metabolic features with SL1344, with several notable differences in the Voges-Proskauer and catalase tests, as well alterations in melibiose, and inositol utilization. These results represent the first full duration infection study using an ST313 strain following the entire natural course of disease progression, and serve as a benchmark for ongoing and future studies into the pathogenesis of D23580.

In vitro models that mimic in vivo host-pathogen interactions are needed to evaluate candidate drugs that inhibit bacterial virulence traits. We established a new approach to study Pseudomonas aeruginosa biofilm susceptibility on biotic surfaces, using a three-dimensional (3-D) lung epithelial cell model. P. aeruginosa formed antibiotic resistant biofilms on 3-D cells without affecting cell viability. The biofilm-inhibitory activity of antibiotics and/or the anti-biofilm peptide DJK-5 were evaluated on 3-D cells compared to a plastic surface, in medium with and without fetal bovine serum (FBS). In both media, aminoglycosides were more efficacious in the 3-D cell model. In serum-free medium, most antibiotics (except polymyxins) showed enhanced efficacy when 3-D cells were present. In medium with FBS, colistin was less efficacious in the 3-D cell model. DJK-5 exerted potent inhibition of P. aeruginosa association with both substrates, only in serum-free medium. DJK-5 showed stronger inhibitory activity against P. aeruginosa associated with plastic compared to 3-D cells. The combined addition of tobramycin and DJK-5 exhibited more potent ability to inhibit P. aeruginosa association with both substrates. In conclusion, lung epithelial cells influence the efficacy of most antimicrobials against P. aeruginosa biofilm formation, which in turn depends on the presence or absence of FBS.

Despite serving as the primary entry portal for coxsackievirus B (CVB), little is known about CVB infection of the intestinal epithelium, owing at least in part to the lack of suitable in vivo models and the inability of cultured cells to recapitulate the complexity and structure associated with the gastrointestinal (GI) tract. Here, we report on the development of a three-dimensional (3-D) organotypic cell culture model of Caco-2 cells to model CVB infection of the gastrointestinal epithelium. We show that Caco-2 cells grown in 3-D using the rotating wall vessel (RWV) bioreactor recapitulate many of the properties of the intestinal epithelium, including the formation of well-developed tight junctions, apical-basolateral polarity, brush borders, and multicellular complexity. In addition, transcriptome analyses using transcriptome sequencing (RNA-Seq) revealed the induction of a number of genes associated with intestinal epithelial differentiation and/or intestinal processes in vivo when Caco-2 cells were cultured in 3-D. Applying this model to CVB infection, we found that although the levels of intracellular virus production were similar in two-dimensional (2-D) and 3-D Caco-2 cell cultures, the release of infectious CVB was enhanced in 3-D cultures at early stages of infection. Unlike CVB, the replication of poliovirus (PV) was significantly reduced in 3-D Caco-2 cell cultures. Collectively, our studies show that Caco-2 cells grown in 3-D using the RWV bioreactor provide a cell culture model that structurally and transcriptionally represents key aspects of cells in the human GI tract and can thus be used to expand our understanding of enterovirus-host interactions in intestinal epithelial cells.

In eutherians, the placenta acts as a barrier and conduit at the maternal-fetal interface. Syncytiotrophoblasts, the multinucleated cells that cover the placental villous tree surfaces of the human placenta, are directly bathed in maternal blood and are formed by the fusion of progenitor cytotrophoblasts that underlie them. Despite their crucial role in fetal protection, many of the events that govern trophoblast fusion and protection from microbial infection are unknown. We describe a three-dimensional (3D)–based culture model using human JEG-3 trophoblast cells that develop syncytiotrophoblast phenotypes when cocultured with human microvascular endothelial cells. JEG-3 cells cultured in this system exhibit enhanced fusogenic activity and morphological and secretory activities strikingly similar to those of primary human syncytiotrophoblasts. RNASeq analyses extend the observed functional similarities to the transcriptome, where we observed significant overlap between syncytiotrophoblast-specific genes and 3D JEG-3 cultures. Furthermore, JEG-3 cells cultured in 3D are resistant to infection by viruses and Toxoplasma gondii, which mimics the high resistance of syncytiotrophoblasts to microbial infections in vivo. Given that this system is genetically manipulatable, it provides a new platform to dissect the mechanisms involved in syncytiotrophoblast development and microbial resistance.

Attributing observed CO2 variations to human or natural cause is critical to deducing and tracking emissions from observations. We have used in situ CO2, CO, and planetary boundary layer height (PBLH) measurements recorded during the CalNex-LA (CARB et al., 2008) ground campaign of 15 May-15 June 2010, in Pasadena, CA, to deduce the diurnally varying anthropogenic component of observed CO2 in the megacity of Los Angeles (LA). This affordable and simple technique, validated by carbon isotope observations and WRF-STILT (Weather Research and Forecasting model - Stochastic Time-Inverted Lagrangian Transport model) predictions, is shown to robustly attribute observed CO2 variation to anthropogenic or biogenic origin over the entire diurnal cycle. During CalNex-LA, local fossil fuel combustion contributed up to similar to 50% of the observed CO2 enhancement overnight, and similar to 100% of the enhancement near midday. This suggests that sufficiently accurate total column CO2 observations recorded near midday, such as those from the GOSAT or OCO-2 satellites, can potentially be used to track anthropogenic emissions from the LA megacity.