Reactivation of M. tuberculosis Infection in Trans-Membrane Tumour Necrosis Factor Mice

Of those individuals who are infected with M. tuberculosis, 90% do not develop active disease and represents a large reservoir of M. tuberculosis with the potential for reactivation of infection. Sustained TNF expression is required for containment of persistent infection and TNF neutralization leads to tuberculosis reactivation. In this study, we investigated the contribution of soluble TNF (solTNF) and transmembrane TNF (Tm-TNF) in immune responses generated against reactivating tuberculosis. In a chemotherapy induced tuberculosis reactivation model, mice were challenged by aerosol inhalation infection with low dose M. tuberculosis for three weeks to establish infection followed chemotherapeutic treatment for six weeks, after which therapy was terminated and tuberculosis reactivation investigated. We demonstrate that complete absence of TNF results in host susceptibility to M. tuberculosis reactivation in the presence of established mycobacteria-specific adaptive immunity with mice displaying unrestricted bacilli growth and diffused granuloma structures compared to WT control mice. Interestingly, bacterial re-emergence is contained in Tm-TNF mice during the initial phases of tuberculosis reactivation, indicating that Tm-TNF sustains immune pressure as in WT mice. However, Tm-TNF mice show susceptibility to long term M. tuberculosis reactivation associated with uncontrolled influx of leukocytes in the lungs and reduced IL-12p70, IFNγ and IL-10, enlarged granuloma structures, and failure to contain mycobacterial replication relative to WT mice. In conclusion, we demonstrate that both solTNF and Tm-TNF are required for maintaining immune pressure to contain reactivating M. tuberculosis bacilli even after mycobacteria-specific immunity has been established

PhotoATRP Induced Self-Assembly (PhotoATR-PISA) Enables Simplified Synthesis of Responsive Polymer Nanoparticles in One-Pot

ABSTRACT: Photo-controlled atom transfer radical polymerization (PhotoATRP) was implemented, for the first time, to accomplish polymerization induced self-assembly (PISA) mediated by UV light (λ = 365 nm) using ppm levels (ca. < 20 ppm) of copper catalyst at ambient temperature. Using CuIIBr2/tris(pyridin-2-ylmethyl)amine (TPMA) catalyst systems, PISA was per-formed all in one-pot starting from synthesis of solvophilic poly(oligo(ethylene oxide) methyl ether methacrylate) (POEGMA) blocks to core-crosslinked nanoparticles (NPs) utilizing poly(glycidyl methacrylate) (PGMA) and N,N-cystamine bismethacrylamide (CBMA) as the solvophobic copolymer and crosslinking agent, respectively. Sequential chain-extensions were performed for PGMA demonstrating capabilities for accessing multi-block copolymers with temporal control via switching the UV light on and off. Further, core-crosslinking of PISA nanoparticles was performed via the slow incorporation of the CBMA enabling one-pot crosslinking during the PISA process. Finally, the disulfide installed in the CBMA core-crosslinks allowed for the stimuli-triggered dissociation of nanoparticles using DL-dithiothreitol at acidic pH.</p

Asymmetric Polymerization-Induced Crystallization-Driven Self-Assembly of Helical, Rod-Coil Poly(Aryl Isocyanide) Block Copolymers

The helical motif is ubiquitous in naturally occurring chemical systems where it confers numerous useful and distinct material properties. Accordingly, a great many synthetic materials have been produced in efforts to mimic biological systems. As an inherently chiral structural feature, the helix provides a modular platform for the creation of chiral nanomaterials exhibiting a diverse range of capabilities. Nonetheless, the development of rapid, scalable methods for production of chiral nanomaterials presents a formidable challenge. However, recent advances in both amphiphilic block copolymer (BCP) synthesis and self-assembly provide tractable means to do just that. In this work, polymerization induced crystallization-driven self-assembly (PI-CDSA) is combined, for the first time, with helical, rod-coil BCP self-assembly to enable scalable and controllable in situ synthesis of chiral nanostructures with variable shape, size, and dimensionality. Herein, we detail the use of newly developed Asymmetric PI-CDSA methodologies in the synthesis and in situ self-assembly of chiral, rod-coil BCPs comprised of poly(aryl isocyanide) (PAIC) rigid-rod and poly(ethylene glycol) (PEG) random-coil components. Using PEG-based Ni(II) macroinitiators, chiral PAIC-BCP nanostructures are constructed in a controlled, scalable fashion forming variable chiral morphologies including 1D twisted nanofibers, 2D hexagonal nanosheets and 3D twisted spirangles (i.e., spirally arranged hexagonal nanosheet stacks). Using seeded, living asymmetric PI-CDSA, the lengths and heights of 1D nanofiber and 3D spirangle nanostructures, respectively, can be selectively tuned via alterations in unimer-to-seed ratios. The formation of these nanostructures is dictated by the liquid crystalline nature of PAIC blocks and the hierarchical assembly of these BCPs, with chirality translated across length scales and in multiple dimensions (i.e., spirangles), led to large amplifications in chiroptical activity with high Kuhn’s dissymmetry factors reaching 0.029

A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds

Historically, microbes from the environment have been a reliable source for novel bio-active compounds. Cloning and expression of metagenomic DNA in heterologous strains of bacteria has broadened the range of potential compounds accessible. However, such metagenomic libraries have been under-exploited for applications in mammalian cells because of a lack of integrated methods. We present an innovative platform to systematically mine natural resources for pro-apoptotic compounds that relies on the combination of bacterial delivery and droplet microfluidics. Using the violacein operon from C. violaceum as a model, we demonstrate that E. coli modified to be invasive can serve as an efficient delivery vehicle of natural compounds. This approach permits the seamless screening of metagenomic libraries with mammalian cell assays and alleviates the need for laborious extraction of natural compounds. In addition, we leverage the unique properties of droplet microfluidics to amplify bacterial clones and perform clonal screening at high-throughput in place of one-compound-per-well assays in multi-well format. We also use droplet microfluidics to establish a cell aggregate strategy that overcomes the issue of background apoptosis. Altogether, this work forms the foundation of a versatile platform to efficiently mine the metagenome for compounds with therapeutic potential

A droplet microfluidics based platform for mining metagenomic libraries for natural compounds

Historically, microbes from the environment have been a reliable source for novel bio-active compounds. Cloning and expression of metagenomic DNA in heterologous strains of bacteria has broadened the range of potential compounds accessible. However, such metagenomic libraries have been under-exploited for applications in mammalian cells because of a lack of integrated methods. We present an innovative platform to systematically mine natural resources for pro-apoptotic compounds that relies on the combination of bacterial delivery and droplet microfluidics. Using the violacein operon from C. violaceum as a model, we demonstrate that E. coli modified to be invasive can serve as an efficient delivery vehicle of natural compounds. This approach permits the seamless screening of metagenomic libraries with mammalian cell assays and alleviates the need for laborious extraction of natural compounds. In addition, we leverage the unique properties of droplet microfluidics to amplify bacterial clones and perform clonal screening at high-throughput in place of one-compound-per-well assays in multi-well format. We also use droplet microfluidics to establish a cell aggregate strategy that overcomes the issue of background apoptosis. Altogether, this work forms the foundation of a versatile platform to efficiently mine the metagenome for compounds with therapeutic potential. ©2017 Keywords: metagenomic screening; droplet microfluidics; high-throughput screening; natural compoundNational Institute of Health (grat no. NCI-1R43GM95227

Dynamic Worm-Gel Materials as Tunable, Regenerable Adsorbents for Water Treatment

Introducing facile regenerability into adsorbent materials can potentially increase sustainability in water treatment systems enabled by extended use. Herein, we detail our recent syntheses of dynamic, nanostructured worm-gel materials and their implementation as regenerable adsorbents for water treatment. Photo-controlled atom transfer radical polymerization-induced self-assembly (PhotoATR-PISA) was employed to synthesize various polymer nanstructures, including dispersed spheres, worms, and vesicles, and nanostructured worm-gels, via the synthesis and simultaneous, in situ assembly of BAB triblock copolymers. Two dynamic, disulfide-functionalized macroinitiators (SS-MI-1 and 2) with different degree of polymerization and one non-dynamic macroinitiator (CC-MI) were synthesized via polymerization of oligo(ethylene glycol methyl ether methacrylate) (OEGMA). PhotoATR-PISA was then implemented via the chain extension from SS-MI-1, 2 and CC-MI with glycidyl methacrylate (GMA) or benzyl methacrylate (BMA) forming BAB-type triblock copolymer nanoparticles in situ. The final morphology in PhotoATR-PISA was influenced not only by conventional factors such as solids content and block DP but also by unimer exchange rates yielding arrested, nanostructured worm-gels in many instances and arrested vesicle-gels in one instance. These PISA-gel materials were implemented as adsorbents for phenanthrene, a model compound registered as a priority pollutant by US EPA, from aqueous solutions. The chemical tunability of these materials enabled enhanced, targeted removal of phenanthrene facilitated by π-π interactions as evidenced by the increased adsorption capacities of PBMA-based PISA-gels when compared to PGMA. Furthermore, the dynamicity of disulfide worm-gels (SS-WG) enabled disulfide exchange-induced regeneration stimulated by UV light. This UV-responsive exchange was investigated for POEGMA macroinitiators as well as dissolved triblock copolymers, dispersed nanoparticles and SS-WG materials. Finally, the regenerability of the PNT-saturated SS-WG adsorbents induced by UV irradiation (λ = 365 nm) was examined and compared to control worm-gels without disulfides demonstrating enhanced recovery of adsorption capacity under mild irradiation conditions

Are rare species useful species? Obstacles to the conservation of tree diversity in the dry forest zone agro-ecosystems of Mesoamerica

Aim To test the potential to conserve rare dry forest tree and shrub species circa situm.Location Oaxaca, Mexico and Southern Honduras.Methods Local uses (timber, posts and firewood) of species were determined principally through semistructured interviews with 20 rural householders in each of four communities in Honduras and four in Oaxaca. Tree and shrub diversity inventories were carried out in a total of 227 forest patches and parcels of farmland in those eight communities. Species’ conservation priorities were determined using the star system of Hawthorne (1996) and IUCN listings.Results Despite a large number of useful species, remarkably few were also conservation priorities. Useful species were found to be substitutable as is illustrated by Bombacopsis quinata, Cordia alliodora, Guaiacum sanctum and G. coulteri.Conclusions In these areas, circa situm conservation is inhibited by the lack of species that are both rare and useful. Usefulness must be interpreted as a function of substitutability. Natural regeneration provides an abundance of diversity, farmers are unlikely to invest in the management of a species when suitable substitutes are freely available. The key to conserving rare species may be in maintaining or enhancing the value of the landscape elements in which they are found

BioTIME:a database of biodiversity time series for the Anthropocene

Abstract Motivation: The BioTIME database contains raw data on species identities and abundances in ecological assemblages through time. These data enable users to calculate temporal trends in biodiversity within and amongst assemblages using a broad range of metrics. BioTIME is being developed as a community‐led open‐source database of biodiversity time series. Our goal is to accelerate and facilitate quantitative analysis of temporal patterns of biodiversity in the Anthropocene. Main types of variables included: The database contains 8,777,413 species abundance records, from assemblages consistently sampled for a minimum of 2 years, which need not necessarily be consecutive. In addition, the database contains metadata relating to sampling methodology and contextual information about each record. Spatial location and grain: BioTIME is a global database of 547,161 unique sampling locations spanning the marine, freshwater and terrestrial realms. Grain size varies across datasets from 0.0000000158 km² (158 cm²) to 100 km² (1,000,000,000,000 cm²). Time period and grain: BioTIME records span from 1874 to 2016. The minimal temporal grain across all datasets in BioTIME is a year. Major taxa and level of measurement: BioTIME includes data from 44,440 species across the plant and animal kingdoms, ranging from plants, plankton and terrestrial invertebrates to small and large vertebrates. Software format: .csv and .SQL