The probiotic effects of Lactobacillus reuteri have been speculated to partly depend on its capacity to produce the antimicrobial substance reuterin during the reduction of glycerol in the gut. In this study, the potential of this process to protect human intestinal epithelial cells against infection with Salmonella enterica serovar Typhimurium was investigated. We used a three-dimensional (3-D) organotypic model of human colonic epithelium that was previously validated and applied to study interactions between S. Typhimurium and the intestinal epithelium that lead to enteric salmonellosis. Using this model system, we show that L. reuteri protects the intestinal cells against the early stages of Salmonella infection and that this effect is significantly increased when L. reuteri is stimulated to produce reuterin from glycerol. More specifically, the reuterin-containing ferment of L. reuteri caused a reduction in Salmonella adherence and invasion (1 log unit), and intracellular survival (2 log units). In contrast, the L. reuteri ferment without reuterin stimulated growth of the intracellular Salmonella population with 1 log unit. The short-term exposure to reuterin or the reuterin-containing ferment had no observed negative impact on intestinal epithelial cell health. However, long-term exposure (24 h) induced a complete loss of cell-cell contact within the epithelial aggregates and compromised cell viability. Collectively, these results shed light on a potential role for reuterin in inhibiting Salmonella-induced intestinal infections and may support the combined application of glycerol and L. reuteri. While future in vitro and in vivo studies of reuterin on intestinal health should fine-tune our understanding of the mechanistic effects, in particular in the presence of a complex gut microbiota, this the first report of a reuterin effect on the enteric infection process in any mammalian cell type.

Background: Buruli ulcer, caused by Mycobacterium ulcerans, is a localized skin lesion that can progress to extensive ulceration and necrosis if left untreated. Unpublished studies of hydrated clays for therapeutic, topical treatment of Buruli ulcer suggest that specific clay mineral products may have beneficial effects on wound healing. In this study, we evaluated the in vitro antibacterial activity of a panel of clay mixtures and their derivative leachates against M. ulcerans and assessed the in vivo efficacy of topically-applied, hydrated clays on Buruli ulcer progression in mice infected with M. ulcerans.

Methods: M. ulcerans 1615 was incubated with 10 % suspensions of CB07, CB08, CB09, CB10, and BY07 clay mixtures, and survival was determined over 28 days. For animal experiments, we examined the effect of topical hydrated clay therapy on Buruli ulcer progression in vivo in mouse tails subcutaneously infected with M. ulcerans 1615.

Results: The CB07, CB08, and CB09 clays exhibited bactericidal activity against M. ulcerans after 7, 14, 21, and 28 days of incubation. In contrast, clay leachates exhibited inhibitory, bacteriostatic effects on M. ulcerans growth in vitro. After establishing an ulcerative M. ulcerans infection for three months, ulcerated regions of the tails were treated once daily (five consecutive days per week) for 22 days with hydrated CB09 clay poultices. Mice in the clay treatment group exhibited healing as assessed by gross morphological changes and a reduction in M. ulcerans present in the wounds.

Conclusions: These data reveal that specific clays exhibit in vitro bactericidal activity against M. ulcerans and that hydrated clay poultices may offer a complementary and integrative strategy for topically treating Buruli ulcer disease.

Background: Salmonella has been employed to deliver therapeutic molecules against cancer and infectious diseases. As the carrier for target gene(s), the cargo plasmid should be stable in the bacterial vector. Plasmid recombination has been reduced in E. coli by mutating several genes including the recA, recE, recF and recJ. However, to our knowledge, there have been no published studies of the effect of these or any other genes that play a role in plasmid recombination in Salmonella enterica.

Results: The effect of recA, recF and recJ deletions on DNA recombination was examined in three serotypes of Salmonella enterica. We found that (1) intraplasmid recombination between direct duplications was RecF-independent in Typhimurium and Paratyphi A, but could be significantly reduced in Typhi by a ΔrecA or ΔrecF mutation; (2) in all three Salmonella serotypes, both ΔrecA and ΔrecF mutations reduced intraplasmid recombination when a 1041 bp intervening sequence was present between the duplications; (3) ΔrecA and ΔrecF mutations resulted in lower frequencies of interplasmid recombination in Typhimurium and Paratyphi A, but not in Typhi; (4) in some cases, a ΔrecJ mutation could reduce plasmid recombination but was less effective than ΔrecA and ΔrecF mutations. We also examined chromosome-related recombination. The frequencies of intrachromosomal recombination and plasmid integration into the chromosome were 2 and 3 logs lower than plasmid recombination frequencies in Rec[superscript +] strains. A ΔrecA mutation reduced both intrachromosomal recombination and plasmid integration frequencies.

Conclusions: The ΔrecA and ΔrecF mutations can reduce plasmid recombination frequencies in Salmonella enterica, but the effect can vary between serovars. This information will be useful for developing Salmonella delivery vectors able to stably maintain plasmid cargoes for vaccine development and gene therapy.

In mammals, transit through the epididymis, which involves the acquisition, loss and modification of proteins, is required to confer motility and fertilization competency to sperm. The overall dynamics of maturation is poorly understood, and a systems level understanding of the complex maturation process will provide valuable new information about changes occurring during epididymal transport. We report the proteomes of sperm collected from the caput, corpus and cauda segments of the mouse epididymis, identifying 1536, 1720 and 1234 proteins respectively. This study identified 765 proteins that are present in sperm obtained from all three segments. We identified 1766 proteins that are potentially added (732) or removed (1034) from sperm during epididymal transit. Phenotypic analyses of the caput, corpus and cauda sperm proteomes identified 60 proteins that have known sperm phenotypes when mutated, or absent from sperm. Our analysis indicates that as much as one-third of proteins with known sperm phenotypes are added to sperm during epididymal transit. GO analyses revealed that cauda sperm are enriched for specific functions including sperm-egg recognition and motility, consistent with the observation that sperm acquire motility and fertilization competency during transit through the epididymis. In addition, GO analyses revealed that the immunity protein profile of sperm changes during sperm maturation. Finally, we identified components of the 26S proteasome, the immunoproteasome, and a proteasome activator in mature sperm.

This study presents the first global transcriptional profiling and phenotypic characterization of the major human opportunistic fungal pathogen, Candida albicans, grown in spaceflight conditions. Microarray analysis revealed that C. albicans subjected to short-term spaceflight culture differentially regulated 452 genes compared to synchronous ground controls, which represented 8.3% of the analyzed ORFs. Spaceflight-cultured C. albicans–induced genes involved in cell aggregation (similar to flocculation), which was validated by microscopic and flow cytometry analysis. We also observed enhanced random budding of spaceflight-cultured cells as opposed to bipolar budding patterns for ground samples, in accordance with the gene expression data. Furthermore, genes involved in antifungal agent and stress resistance were differentially regulated in spaceflight, including induction of ABC transporters and members of the major facilitator family, downregulation of ergosterol-encoding genes, and upregulation of genes involved in oxidative stress resistance.

Finally, downregulation of genes involved in actin cytoskeleton was observed. Interestingly, the transcriptional regulator Cap1 and over 30% of the Cap1 regulon was differentially expressed in spaceflight-cultured C. albicans. A potential role for Cap1 in the spaceflight response of C. albicans is suggested, as this regulator is involved in random budding, cell aggregation, and oxidative stress resistance; all related to observed spaceflight-associated changes of C. albicans. While culture of C. albicans in microgravity potentiates a global change in gene expression that could induce a virulence-related phenotype, no increased virulence in a murine intraperitoneal (i.p.) infection model was observed under the conditions of this study. Collectively, our data represent an important basis for the assessment of the risk that commensal flora could play during human spaceflight missions. Furthermore, since the low fluid-shear environment of microgravity is relevant to physical forces encountered by pathogens during the infection process, insights gained from this study could identify novel infectious disease mechanisms, with downstream benefits for the general public.

The sensitivity of Earth’s wetlands to observed shifts in global precipitation and temperature patterns and their ability to produce large quantities of methane gas are key global change questions. We present a microwave satellite-based approach for mapping fractional surface water (FW) globally at 25-km resolution. The approach employs a land cover-supported, atmospherically-corrected dynamic mixture model applied to 20+ years (1992–2013) of combined, daily, passive/active microwave remote sensing data. The resulting product, known as Surface Water Microwave Product Series (SWAMPS), shows strong microwave sensitivity to sub-grid scale open water and inundated wetlands comprising open plant canopies. SWAMPS’ FW compares favorably (R² = 91%–94%) with higher-resolution, global-scale maps of open water from MODIS and SRTM-MOD44W. Correspondence of SWAMPS with open water and wetland products from satellite SAR in Alaska and the Amazon deteriorates when exposed wetlands or inundated forests captured by the SAR products were added to the open water fraction reflecting SWAMPS’ inability to detect water underneath the soil surface or beneath closed forest canopies. Except for a brief period of drying during the first 4 years of observation, the inundation extent for the global domain excluding the coast was largely stable. Regionally, inundation in North America is advancing while inundation is on the retreat in Tropical Africa and North Eurasia. SWAMPS provides a consistent and long-term global record of daily FW dynamics, with documented accuracies suitable for hydrologic assessment and global change-related investigations.

Many recent studies observe the increasing importance, influence, and analysis of resilience as a concept to understand the capacity of a system or individual to respond to change. The term has achieved prominence in diverse scientific fields, as well as public discourse and policy arenas. As a result, resilience has been referred to as a boundary object or a bridging concept that is able to facilitate communication and understanding across disciplines, coordinate groups of actors or stakeholders, and build consensus around particular policy issues. We present a network analysis of bibliometric data to understand the extent to which resilience can be considered as a boundary object or a bridging concept in terms of its links across disciplines and scientific fields. We analyzed 994 papers and 35,952 citations between them to reveal the connectedness and links between and within fields. We analyzed the network according to different fields, modules, and sub-fields, showing a highly clustered citation network. Analyzing betweenness allowed us to identify how particular papers bridge across fields and how different fields are linked. With the exception of a few specific papers, most papers cite exclusively within their own field. We conclude that resilience is to an extent a boundary object because there are shared understandings across diverse disciplines and fields. However, it is more limited as a bridging concept because the citations across fields are concentrated among particular disciplines and papers, so the distinct fields do not widely or routinely refer to each other. There are some signs of resilience being used as an interdisciplinary concept to bridge scientific fields, particularly in social-ecological systems, which may itself constitute an emerging sub-field.

A warming climate is altering land-atmosphere exchanges of carbon, with a potential for increased vegetation productivity as well as the mobilization of permafrost soil carbon stores. Here we investigate land-atmosphere carbon dioxide (CO₂) cycling through analysis of net ecosystem productivity (NEP) and its component fluxes of gross primary productivity (GPP) and ecosystem respiration (ER) and soil carbon residence time, simulated by a set of land surface models (LSMs) over a region spanning the drainage basin of Northern Eurasia. The retrospective simulations cover the period 1960–2009 at 0.5° resolution, which is a scale common among many global carbon and climate model simulations. Model performance benchmarks were drawn from comparisons against both observed CO₂ fluxes derived from site-based eddy covariance measurements as well as regional-scale GPP estimates based on satellite remote-sensing data.

The site-based comparisons depict a tendency for overestimates in GPP and ER for several of the models, particularly at the two sites to the south. For several models the spatial pattern in GPP explains less than half the variance in the MODIS MOD17 GPP product. Across the models NEP increases by as little as 0.01 to as much as 0.79 g C m^-2 yr^-2, equivalent to 3 to 340 % of the respective model means, over the analysis period. For the multimodel average the increase is 135 % of the mean from the first to last 10 years of record (1960–1969 vs. 2000–2009), with a weakening CO₂ sink over the latter decades. Vegetation net primary productivity increased by 8 to 30 % from the first to last 10 years, contributing to soil carbon storage gains. The range in regional mean NEP among the group is twice the multimodel mean, indicative of the uncertainty in CO₂ sink strength.

The models simulate that inputs to the soil carbon pool exceeded losses, resulting in a net soil carbon gain amid a decrease in residence time. Our analysis points to improvements in model elements controlling vegetation productivity and soil respiration as being needed for reducing uncertainty in land-atmosphere CO₂ exchange. These advances will require collection of new field data on vegetation and soil dynamics, the development of benchmarking data sets from measurements and remote-sensing observations, and investments in future model development and intercomparison studies.

Astronauts are exposed to a unique combination of stressors during spaceflight, which leads to alterations in their physiology and potentially increases their susceptibility to disease, including infectious diseases. To evaluate the potential impact of the spaceflight environment on the regulation of molecular pathways mediating cellular stress responses, we performed a first-of-its-kind pilot study to assess spaceflight-related gene-expression changes in the whole blood of astronauts. Using an array comprised of 234 well-characterized stress-response genes, we profiled transcriptomic changes in six astronauts (four men and two women) from blood preserved before and immediately following the spaceflight. Differentially regulated transcripts included those important for DNA repair, oxidative stress, and protein folding/degradation, including HSP90AB1, HSP27, GPX1, XRCC1, BAG-1, HHR23A, FAP48, and C-FOS. No gender-specific differences or relationship to number of missions flown was observed. This study provides a first assessment of transcriptomic changes occurring in the whole blood of astronauts in response to spaceflight.

Non-typhoidal Salmonella are associated with gastrointestinal disease worldwide and invasive disease in Africa. We constructed novel bivalent vaccines through the recombinant expression of heterologous O-antigens from Salmonella Choleraesuis in Salmonella Typhimurium. A recombinant Asd⁺ plasmid pCZ1 with the cloned Salmonella Choleraesuis O-antigen gene cluster was introduced into three constructed Salmonella Typhimurium Δasd mutants: SLT11 (ΔrfbP), SLT12 (ΔrmlB-rfbP) and SLT16 (ΔrfbP ∆pagL::TT araCP_BADrfbP). Immunoblotting demonstrated that SLT11 (pCZ1) and SLT12 (pCZ1) efficiently expressed the heterologous O-antigen. In the presence of arabinose, SLT16 (pCZ1) expressed both the homologous and heterologous O-antigens, whereas in the absence of arabinose, SLT16 (pCZ1) mainly expressed the heterologous O-antigen. We deleted the crp/cya genes in SLT12 (pCZ1) and SLT16 (pCZ1) for attenuation purposes, generating the recombinant vaccine strains SLT17 (pCZ1) and SLT18 (pCZ1). Immunization with either SLT17 (pCZ1) or SLT18 (pCZ1) induced specific IgG against the heterologous O-antigen, which mediated significant killing of Salmonella Choleraesuis and provided full protection against a lethal homologous challenge in mice. Furthermore, SLT17 (pCZ1) or SLT18 (pCZ1) immunization resulted in 83% or 50% heterologous protection against Salmonella Choleraesuis challenge, respectively. Our study demonstrates that heterologous O-antigen expression is a promising strategy for the development of multivalent Salmonella vaccines.