Technical assessment of Mir-1 life support hardware for the international space station

NASA has been progressively learning the design and performance of the Russian life support systems utilized in their Mir space station. In 1992, a plan was implemented to assess the benefits of the Mir-1 life support systems to the Freedom program. Three primary tasks focused on: evaluating the operational Mir-1 support technologies and understanding if specific Russian systems could be directly utilized on the American space station and if Russian technology design information could prove useful in improving the current design of the planned American life support equipment; evaluating the ongoing Russian life support technology development activities to determine areas of potential long-term application to the U.S. space station; and utilizing the expertise of their space station life support systems to evaluate the benefits to the current U.S. space station program which included the integration of the Russian Mir-1 designs with the U.S. designs to support a crew of six

Advancing the understanding of treponemal disease in the past and present

Syphilis was perceived to be a new disease in Europe in the late 15th century, igniting a debate about its origin that continues today in anthropological, historical, and medical circles. We move beyond this age-old debate using an interdisciplinary approach that tackles broader questions to advance the understanding of treponemal infection (syphilis, yaws, bejel, and pinta). How did the causative organism(s) and humans co-evolve? How did the related diseases caused by Treponema pallidum emerge in different parts of the world and affect people across both time and space? How are T. pallidum subspecies related to the treponeme causing pinta? The current state of scholarship in specific areas is reviewed with recommendations made to stimulate future work. Understanding treponemal biology, genetic relationships, epidemiology, and clinical manifestations is crucial for vaccine development today and for investigating the distribution of infection in both modern and past populations. Paleopathologists must improve diagnostic criteria and use a standard approach for recording skeletal lesions on archaeological human remains. Adequate contextualization of cultural and environmental conditions is necessary, including site dating and justification for any corrections made for marine or freshwater reservoir effects. Biogeochemical analyses may assess aquatic contributions to diet, physiological changes arising from treponemal disease and its treatments (e.g., mercury), or residential mobility of those affected. Shifting the focus from point of origin to investigating who is affected (e.g., by age/sex or socioeconomic status) and disease distribution (e.g., coastal/ inland, rural/urban) will advance our understanding of the treponemal disease and its impact on people through time

Mineral Type and Solution Chemistry Affect the Structure and Composition of Actively Growing Bacterial Communities as Revealed by Bromodeoxyuridine Immunocapture and 16S rRNA Pyrosequencing

© 2016, Springer Science+Business Media New York. Understanding how minerals affect bacterial communities and their in situ activities in relation to environmental conditions are central issues in soil microbial ecology, as minerals represent essential reservoirs of inorganic nutrients for the biosphere. To determine the impact of mineral type and solution chemistry on soil bacterial communities, we compared the diversity, composition, and functional abilities of a soil bacterial community incubated in presence/absence of different mineral types (apatite, biotite, obsidian). Microcosms were prepared containing different liquid culture media devoid of particular essential nutrients, the nutrients provided only in the introduced minerals and therefore only available to the microbial community through mineral dissolution by biotic and/or abiotic processes. By combining functional screening of bacterial isolates and community analysis by bromodeoxyuridine DNA immunocapture and 16S rRNA gene pyrosequencing, we demonstrated that bacterial communities were mainly impacted by the solution chemistry at the taxonomic level and by the mineral type at the functional level. Metabolically active bacterial communities varied with solution chemistry and mineral type. Burkholderia were significantly enriched in the obsidian treatment compared to the biotite treatment and were the most effective isolates at solubilizing phosphorous or mobilizing iron, in all the treatments. A detailed analysis revealed that the 16S rRNA gene sequences of the OTUs or isolated strains assigned as Burkholderia in our study showed high homology with effective mineral-weathering bacteria previously recovered from the same experimental site

Age-Dependent TLR3 Expression of the Intestinal Epithelium Contributes to Rotavirus Susceptibility

Rotavirus is a major cause of diarrhea worldwide and exhibits a pronounced small intestinal epithelial cell (IEC) tropism. Both human infants and neonatal mice are highly susceptible, whereas adult individuals remain asymptomatic and shed only low numbers of viral particles. Here we investigated age-dependent mechanisms of the intestinal epithelial innate immune response to rotavirus infection in an oral mouse infection model. Expression of the innate immune receptor for viral dsRNA, Toll-like receptor (Tlr) 3 was low in the epithelium of suckling mice but strongly increased during the postnatal period inversely correlating with rotavirus susceptibility, viral shedding and histological damage. Adult mice deficient in Tlr3 (Tlr3−/−) or the adaptor molecule Trif (TrifLps2/Lps2) exerted significantly higher viral shedding and decreased epithelial expression of proinflammatory and antiviral genes as compared to wild-type animals. In contrast, neonatal mice deficient in Tlr3 or Trif did not display impaired cell stimulation or enhanced rotavirus susceptibility. Using chimeric mice, a major contribution of the non-hematopoietic cell compartment in the Trif-mediated antiviral host response was detected in adult animals. Finally, a significant age-dependent increase of TLR3 expression was also detected in human small intestinal biopsies. Thus, upregulation of epithelial TLR3 expression during infancy might contribute to the age-dependent susceptibility to rotavirus infection

Innate Immune Sensing of Modified Vaccinia Virus Ankara (MVA) Is Mediated by TLR2-TLR6, MDA-5 and the NALP3 Inflammasome

Modified vaccinia virus Ankara (MVA) is an attenuated double-stranded DNA poxvirus currently developed as a vaccine vector against HIV/AIDS. Profiling of the innate immune responses induced by MVA is essential for the design of vaccine vectors and for anticipating potential adverse interactions between naturally acquired and vaccine-induced immune responses. Here we report on innate immune sensing of MVA and cytokine responses in human THP-1 cells, primary human macrophages and mouse bone marrow-derived macrophages (BMDMs). The innate immune responses elicited by MVA in human macrophages were characterized by a robust chemokine production and a fairly weak pro-inflammatory cytokine response. Analyses of the cytokine production profile of macrophages isolated from knockout mice deficient in Toll-like receptors (TLRs) or in the adapter molecules MyD88 and TRIF revealed a critical role for TLR2, TLR6 and MyD88 in the production of IFNβ-independent chemokines. MVA induced a marked up-regulation of the expression of RIG-I like receptors (RLR) and the IPS-1 adapter (also known as Cardif, MAVS or VISA). Reduced expression of RIG-I, MDA-5 and IPS-1 by shRNAs indicated that sensing of MVA by RLR and production of IFNβ and IFNβ-dependent chemokines was controlled by the MDA-5 and IPS-1 pathway in the macrophage. Crosstalk between TLR2-MyD88 and the NALP3 inflammasome was essential for expression and processing of IL-1β. Transcription of the Il1b gene was markedly impaired in TLR2−/− and MyD88−/− BMDM, whereas mature and secreted IL-1β was massively reduced in NALP3−/− BMDMs or in human THP-1 macrophages with reduced expression of NALP3, ASC or caspase-1 by shRNAs. Innate immune sensing of MVA and production of chemokines, IFNβ and IL-1β by macrophages is mediated by the TLR2-TLR6-MyD88, MDA-5-IPS-1 and NALP3 inflammasome pathways. Delineation of the host response induced by MVA is critical for improving our understanding of poxvirus antiviral escape mechanisms and for designing new MVA vaccine vectors with improved immunogenicity

High-Affinity Capture of Proteins by Diamond Nanoparticles for Mass Spectrometric Analysis

Carboxylated/oxidized diamond nanoparticles (nominal size 100 nm) exhibit exceptionally high affinity for proteins through both hydrophilic and hydrophobic forces. The affinity is so high that proteins in dilute solution can be easily captured by diamonds, simply separated by centrifugation, and directly analyzed by matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS). No preseparation of the adsorbed molecules from diamonds is required for the mass spectrometric analysis. Compared to conventional MALDI-TOF-MS, an enhancement in detection sensitivity by more than 2 orders of magnitude is achieved for dilute solution containing cytochrome c, myoglobin, and albumin because of preconcentration of the probed molecules. The lowest concentration detectable is 100 pM for a 1-mL solution. Aside from the enhanced sensitivity, the overall performance of this technique does not show any sign of deterioration for highly contaminated protein solutions, and furthermore, no significant peak broadening and band shift were observed in the mass spectra. The promise of this new method for clinical proteomics research is demonstrated with an application to human blood serum. Matrix-assisted laser desorption/ionization (MALDI) 1 time-offlight (TOF) mass spectrometry (MS) is a mainstream tool in current high-throughput mass analysis of biopolymers. 2 The MALDI technique, however, suffers from the shortcoming that it lacks sample specificity and its performance deteriorates markedly for samples containing multiple components and excessive amounts of salts or surfactants. 3 Surface-enhanced laser desorption/ ionization (SELDI) is one of the techniques 4-10 developed to circumvent these problems. In this method, 4 micrometer-sized (typically 80-300 µm in diameter) agarose beads made for affinity chromatography columns were used to capture proteins of interest in crude sample solutions. The microbeads were then recovered, washed, placed on the LDI probe tip, and analyzed with regular MALDI-TOF-MS. Unfortunately, direct analysis of the surfacebound proteins is often accompanied with undesired decrease in mass resolution as well as mass accuracy ascribed to the interference from the beads in ion formation and extraction. One solution to this problem is to directly immobilize proteins onto the surface of the LDI probe without use of the microbeads. 7 The approach again suffers from the shortcoming that the number of binding sites is quite limited, ∼1 × 10 13 molecules/cm 2 or ∼160 fmol/mm 2 for a single layer of proteins on the probe surface. The obstacle was later removed by immobilization of the proteins to high molecular weight dextrans precoated covalently on the LDI probe. 8 An approximate 500 times more sample could be loaded, although the dextran immobilization process is rather timeconsuming. We have previously shown 11 that diamond is an exceptional platform for protein adsorption and immobilization. The optical transparency, chemical inertness, and biological compatibility of the material endow diamond nanoparticles with novel and promising biotechnological applications. Preliminary tests with cytochrome c physisorbed to carboxylated/oxidized diamond particles of 5 and 100 nm in size indicate that the specially prepared diamond surfaces exhibit remarkably high affinity for proteins containing amino acid residues with basic side chains. This unique feature along with the fact that diamond is optically transparent up to the UV region motivated us to explore the possibility of using diamond nanoparticles for SELDI-TOF-MS. The advantage of using nanoparticles over microbeads is manyfold. First, nanoparticles have a much larger surface area-to-mass ratio, nearly 3 orders of magnitude higher than that of microbeads; second, the extent to which nanoparticles interfere with the laser desorption/ ionization process is diminished because of the smallness of the particles; third, nanoparticles can be embedded more firmly in the LDI matrix crystals than microbeads, thereby reducing material loss during sample preparation and analysis. There have been several applications of metallic, semiconducting as well as polymeric nanoparticles for mass spectrometric analysis of biopoly