Building a Robust Commercial Microgravity Economy in Earth's Orbit: Economic Readiness Considerations

The reduced gravity environment of space provides a unique opportunity to further our understanding of various materials phenomena involving the molten, fluidic and gaseous states as well as life science applications where, contrary to earlier beliefs, microgravity induces changes in single cells and simple organisms; not only in large organisms with a complex overall response to gravity (or lack thereof). The potential breadth of commercial opportunities in microgravity thus spans over many verticals of the private sector with applications ranging from fiber optics, high-resolution crystals, microencapsulation, 3D organs to perfume and color dyes. Overall, products manufactured in microgravity hold the promise to have key properties surpassing their best terrestrial counterparts. Commercialization, also known as taking a new technology to market, is a journey in itself where the business, economic, market and technological components must align to generate a successful outcome. A business perspective is very different than technology maturation. In order for a technology to be ready for commercialization, it must not only be mature, but it must also have a compelling business case, and the means to scale up production must be identified and practical. Creating a robust economy in Earths orbit (Fig 1) is especially challenging because of the complexity (high risks, lack of standardization) involved in predicting future growth. This complexity can easily overwhelm the fact that many of the products have an attractive touch of space which aids with branding and marketing.This paper reviews the types of added value that can be extracted from space, with an emphasis on the microgravity environment. In addition, lessons learned from past commercialization efforts will be reviewed. While past efforts have yielded some point successes, they have as a whole failed to precipitate a sustainable LEO based marke

MicroRNA regulation of bovine monocyte inflammatory and metabolic networks in an in vivo infection model.

peer-reviewedBovine mastitis is an inflammation-driven disease of the bovine mammary gland that costs the global dairy industry several billion dollars per annum. Because disease susceptibility is a multi-factorial complex phenotype, an integrative biology approach is required to dissect the molecular networks involved. Here, we report such an approach, using next generation sequencing combined with advanced network and pathway biology methods to simultaneously profile mRNA and miRNA expression at multiple time-points (0, 12, 24, 36 and 48h) in both milk and blood FACS-isolated CD14+ monocytes from animals infected in vivo with Streptococcus uberis. More than 3,700 differentially expressed (DE) genes were identified in milk-isolated monocytes (MIMs), a key immune cell recruited to the site of infection during mastitis. Up-regulated genes were significantly enriched for inflammatory pathways, while down-regulated genes were enriched for non-glycolytic metabolic pathways. Monocyte transcriptional changes in the blood, however, were more subtle but highlighted the impact of this infection systemically. Genes up-regulated in blood-isolated-monocytes (BIMs) showed a significant association with interferon and chemokine signalling. Furthermore, twenty-six miRNAs were differentially expressed in MIMs and three in BIMs. Pathway analysis revealed that predicted targets of down-regulated miRNAs were highly enriched for roles in innate immunity (FDR < 3.4E-8) in particular TLR signalling, while up-regulated miRNAs preferentially targeted genes involved in metabolism. We conclude that during S. uberis infection miRNAs are key amplifiers of monocyte inflammatory response networks and repressors of several metabolic pathways.This study was funded in part by Teagasc RMIS 6018 and United States Department of Agriculture ARS funding 3625-32000-102-00. NL is supported by a Teagasc Walsh Fellowship

Bioassays of Compounds with Potential Juvenoid Activity on \u3ci\u3eDrosophila melanogaster\u3c/i\u3e: Juvenile Hormone III, Bisepoxide Juvenile Hormone III, and Methyl Farnesoates

Metabolites of the 6,7,10,11 bisepoxide juvenile hormone III (JHB3), and other potential juvenoids, were tested for juvenile hormone activity using early instar or early stage pupae of Drosophila mela-nogaster. Importantly, methyl farnesoates were tested as they might have JH-like activity on Dipteran juveniles. Larvae were exposed to compounds in medium, or the compounds were applied to white puparia. In the assays employed in the present study, there was no indication for JH activity associ-ated with the metabolites of JHB3. The activity of methyl farnesoate (MF) was higher than that of JH III and far greater than bisepoxide JH III. As opposed to the two endogenous juvenile hormones, methyl farnesoate has weak activity in the white puparial bioassay. When fluorinated forms of me-thyl farnesoate, which is unlikely to be converted to JH, were applied to Drosophila medium to which fly eggs were introduced, there was a high degree of larval mortality, but no evidence of subsequent mortality at the pupal stage. One possible explanation for the results is that methyl farnesoate is active as a hormone in larval stages, but has little activity at the pupal stage where only juvenile hormone has a major effect

Bioassays of Compounds with Potential Juvenoid Activity on \u3ci\u3eDrosophila melanogaster\u3c/i\u3e: Juvenile Hormone III, Bisepoxide Juvenile Hormone III, and Methyl Farnesoates

Metabolites of the 6,7,10,11 bisepoxide juvenile hormone III (JHB3), and other potential juvenoids, were tested for juvenile hormone activity using early instar or early stage pupae of Drosophila mela-nogaster. Importantly, methyl farnesoates were tested as they might have JH-like activity on Dipteran juveniles. Larvae were exposed to compounds in medium, or the compounds were applied to white puparia. In the assays employed in the present study, there was no indication for JH activity associ-ated with the metabolites of JHB3. The activity of methyl farnesoate (MF) was higher than that of JH III and far greater than bisepoxide JH III. As opposed to the two endogenous juvenile hormones, methyl farnesoate has weak activity in the white puparial bioassay. When fluorinated forms of me-thyl farnesoate, which is unlikely to be converted to JH, were applied to Drosophila medium to which fly eggs were introduced, there was a high degree of larval mortality, but no evidence of subsequent mortality at the pupal stage. One possible explanation for the results is that methyl farnesoate is active as a hormone in larval stages, but has little activity at the pupal stage where only juvenile hormone has a major effect

Gene Dosage Effects at the Imprinted Gnas Cluster

Genomic imprinting results in parent-of-origin-dependent monoallelic gene expression. Early work showed that distal mouse chromosome 2 is imprinted, as maternal and paternal duplications of the region (with corresponding paternal and maternal deficiencies) give rise to different anomalous phenotypes with early postnatal lethalities. Newborns with maternal duplication (MatDp(dist2)) are long, thin and hypoactive whereas those with paternal duplication (PatDp(dist2)) are chunky, oedematous, and hyperactive. Here we focus on PatDp(dist2). Loss of expression of the maternally expressed Gnas transcript at the Gnas cluster has been thought to account for the PatDp(dist2) phenotype. But PatDp(dist2) also have two expressed doses of the paternally expressed Gnasxl transcript. Through the use of targeted mutations, we have generated PatDp(dist2) mice predicted to have 1 or 2 expressed doses of Gnasxl, and 0, 1 or 2 expressed doses of Gnas. We confirm that oedema is due to lack of expression of imprinted Gnas alone. We show that it is the combination of a double dose of Gnasxl, with no dose of imprinted Gnas, that gives rise to the characteristic hyperactive, chunky, oedematous, lethal PatDp(dist2) phenotype, which is also hypoglycaemic. However PatDp(dist2) mice in which the dosage of the Gnasxl and Gnas is balanced (either 2∶2 or 1∶1) are neither dysmorphic nor hyperactive, have normal glucose levels, and are fully viable. But PatDp(dist2) with biallelic expression of both Gnasxl and Gnas show a marked postnatal growth retardation. Our results show that most of the PatDp(dist2) phenotype is due to overexpression of Gnasxl combined with loss of expression of Gnas, and suggest that Gnasxl and Gnas may act antagonistically in a number of tissues and to cause a wide range of phenotypic effects. It can be concluded that monoallelic expression of both Gnasxl and Gnas is a requirement for normal postnatal growth and development

Commercial Application of In-Space Assembly

In-Space assembly (ISA) expands the opportunities for cost effective emplacement of systems in space. Currently, spacecraft are launched into space and deploy into their operational configuration through a carefully choreographed sequence of operations. The deployment operation dictates the arrangement of the primary systems on the spacecraft, limiting the ability to take full advantage of launch vehicles volume and mass capability. ISA enables vastly different spacecraft architectures and emplacement scenarios to be achieved, including optimal launch configurations ranging from single launch and assembly to on-orbit aggregation of multiple launches at different orbital locations and times. The spacecraft can be visited at different orbital locations and times to effect expansion and maintenance of an operational capability. To date, the primary application of ISA has been in large programs funded by government organizations, such as the International Space Station. Recently, Space Systems Loral (SSL) led a study funded by the Defense Advanced Research Projects Agency (DARPA), called Dragonfly, to investigate the commercial applicability and economic advantages of ISA. In the study, it was shown that ISA enables SSL to double the capability of a commercial satellite system by taking advantage of alternate packaging approaches for the reflectors. The study included an ultra-light-weight robotic system, derived from Mars manipulator designs, to complete assembly of portions of the antenna system using a tool derived from DARPA orbital express and National Aeronautics and Space Administration (NASA) automated structural assembly experience. The mechanical connector that enables robotic ISA takes advantage of decades of development by NASA from the 1970's to 1980's during the Space Station Freedom program, the precursor to the ISS. The mechanical connector was originally designed for rapid astronaut assembly while also providing a high quality structural connection with linear load deflection response. The paper will discuss the business case for ISA, the general approach taken to exploit on-orbit assembly in the GEO communication satellite market, and the concept of operations associated with the ISA approach, thus laying the foundation for ISA to become an accepted operational approach for commercial in-space operations

Non-Immunogenicity of Overlapping Gag Peptides Pulsed on Autologous Cells after Vaccination of HIV Infected Individuals

Background: HIV Gag-specific CD4+ and CD8+ T-cell responses are important for HIV immune control. Pulsing overlapping Gag peptides on autologous lymphocytes (OPAL) has proven immunogenic and effective in reducing viral loads in multiple pigtail macaque studies, warranting clinical evaluation. Methodology We performed a phase I, single centre, placebo-controlled, double-blinded and dose-escalating study to evaluate the safety and preliminary immunogenicity of a novel therapeutic vaccine approach ‘OPAL-HIV-Gag(c)’. This vaccine is comprised of 120 15mer peptides, overlapping by 11 amino acids, spanning the HIV Gag C clade sequence proteome, pulsed on white blood cells enriched from whole blood using a closed system, followed by intravenous reinfusion. Patients with undetectable HIV viral loads (<50 copies/ml plasma) on HAART received four administrations at week 0, 4, 8 and 12, and were followed up for 12 weeks post-treatment. Twenty-three people were enrolled in four groups: 12 mg (n = 6), 24 mg (n = 7), 48 mg (n = 2) or matching placebo (n = 8) with 18 immunologically evaluable. T-cell immunogenicity was assessed by IFNγ ELIspot and intracellular cytokine staining (ICS). Results: The OPAL-HIV-Gag(c) peptides were antigenic in vitro in 17/17 subjects. After vaccination with OPAL-HIV-Gag(c), 1/6 subjects at 12 mg and 1/6 subjects at 24 mg dose groups had a 2- and 3-fold increase in ELIspot magnitudes from baseline, respectively, of Gag-specific CD8+ T-cells at week 14, compared to 0/6 subjects in the placebo group. No Gag-specific CD4+ T-cell responses or overall change in Rev, Nef, Tat and CMV specific responses were detected. Marked, transient and self-limiting lymphopenia was observed immediately post-vaccination (4 hours) in OPAL-HIV-Gag(c) but not in placebo recipients, with median fall from 1.72 to 0.67 million lymphocytes/mL for active groups (P<0.001), compared to post-placebo from 1.70 to 1.56 lymphocytes/ml (P = 0.16). Conclusion/Significance Despite strong immunogenicity observed in several Macaca nemestrina studies using this approach, OPAL-HIV-Gag(c) was not significantly immunogenic in humans and improved methods of generating high-frequency Gag-specific T-cell responses are required. Name of Registry ClinicalTrials.gov, Registry number: NCT01123915, URL trial registry database: http://www.clinicaltrials.gov/ct2/results?term=OPAL-HIV-1001&Search=Searc