Paper Session I-A - Planning for Operations On-Board the International Space Station

With the launch of the first element of the International Space Station (ISS) in late 1997, scientists and engineers from around the world will have greater access to the space environment for research and commercial exploration. The complexity and flexibility of the International Space Station offers opportunities as well as challenges in planning on-orbit operations. In order to make effective use of limited resources (e.g. crew time, power, and data), while maximizing the results to the scientific, commercial, military and educational communities, the ISS operations planning community must balance many constraints and criteria. The four disciplines must be aware of these constraints as well as the planning process to plan and conduct their activities on-board ISS

Defining the current distribution of the imperiled Black-spotted Newt across south Texas, USA

The Black-spotted Newt (Notophthalmus meridionalis) is a chronically understudied salamander species, with many aspects of its natural history, ecology, and distribution poorly known. Previous studies using traditional methodologies have had limited success documenting N. meridionalis on the landscape, detecting individuals at 6% (7 of 114) and 1% (2 of 221) of sites surveyed. A novel environmental DNA (eDNA) assay was designed and implemented with the goals of assessing the current distribution of N. meridionalis across south Texas, USA, and better understanding the conditions for positive eDNA detections. We conducted eDNA sampling and traditional surveys at 80 sites throughout south Texas. Notophthalmus meridionalis was detected at 12 localities in total: four localities using eDNA surveys, four localities using traditional methods, and four localities with both methodologies. eDNA detections were obtained from five counties, including one where N. meridionalis has never been reported and another where N. meridionalis has not been observed since the 1930s. eDNA detections were obtained in all four seasons, generally following moderate to heavy rainfall events. Our results support the increased use of eDNA surveys to detect rare and cryptic amphibians and to better understand the current distribution of this imperiled species

Measurement of Calcium Dissociation Rates from Troponin C in Rigor Skeletal Myofibrils

Ca2+ dissociation from the regulatory domain of troponin C may influence the rate of striated muscle relaxation. However, Ca2+ dissociation from troponin C has not been measured within the geometric and stoichiometric constraints of the muscle fiber. Here we report the rates of Ca2+ dissociation from the N-terminal regulatory and C-terminal structural domains of fluorescent troponin C constructs reconstituted into rabbit rigor psoas myofibrils using stopped-flow technology. Chicken skeletal troponin C fluorescently labeled at Cys 101, troponin CIAEDANS, reported Ca2+ dissociation exclusively from the structural domain of troponin C at ∼0.37, 0.06, and 0.07/s in isolation, in the presence of troponin I and in myofibrils at 15°C, respectively. Ca2+ dissociation from the regulatory domain was observed utilizing fluorescently labeled troponin C containing the T54C and C101S mutations. Troponin CMIANST54C,C101S reported Ca2+ dissociation exclusively from the regulatory domain of troponin C at >1000, 8.8, and 15/s in isolation, in the presence of troponin I and in myofibrils at 15°C, respectively. Interestingly, troponin CIAANST54C,C101S reported a biphasic fluorescence change upon Ca2+ dissociation from the N- and C-terminal domains of troponin C with rates that were similar to those reported by troponin CMIANST54C,C101S and troponin CIAEDANS at all levels of the troponin C systems. Furthermore, the rate of Ca2+ dissociation from troponin C in the myofibrils was similar to the rate of Ca2+ dissociation measured from the troponin C-troponin I complexes. Since the rate of Ca2+ dissociation from the regulatory domain of TnC in myofibrils is similar to the rate of skeletal muscle relaxation, Ca2+ dissociation from troponin C may influence relaxation

The Effect of Matrix Stiffness of Biomimetic Gelatin Nanofibrous Scaffolds on Human Cardiac Pericyte Behaviour

Congenital heart disease (CHD) is the most common and deadly congenital anomaly, accounting for up to 7.5% of all infant deaths. Survival in children born with CHD has improved dramatically over the past several decades (this positive trend being counterbalanced by the fact that more patients develop heart failure). Seminal data indicate an alteration of the extracellular matrix occurs with time in these hearts due to diffuse and abundant interstitial fibrosis. This results in an escalation in the stiffness of the local myocardial microenvironment. However, the influence of matrix stiffness in regulating the function of resident human stromal cells has not been reported. The objective of this study was to determine the impact of scaffold stiffness on the antigenic and functional profile of cardiac pericytes (CPs) isolated from patients with CHD. To this end, we have first manufactured gelatin nanofibrous scaffolds with varying degrees of stiffness using an in situ cross-linking electrospinning technique in a pure water solvent system. We assessed Young’s modulus and performed a comprehensive physicochemical characterization of the scaffolds employing scanning electron microscopy and Fourier transform infrared spectroscopy. We next evaluated the changes induced by a different scaffold stiffness on CP morphology, antigenic profile, viability, proliferation, angiocrine activity, and induced differentiation. Results indicate that soft matrixes with a fiber diameter of ∼400 nm increase CP proliferation, secretion of angiopoietin 2, and F-actin stress fiber formation, without affecting the antigenic profile, viability, or differentiation. These data indicate for the first time that human CPs can be functionally influenced by slight changes in matrix stiffness. The study elucidates the importance of mechanical/morphological cues in modulating the behavior of stromal cells isolated from patients with CHD

Whole-Genome Sequencing and Concordance Between Antimicrobial Susceptibility Genotypes and Phenotypes of Bacterial Isolates Associated with Bovine Respiratory Disease.

Extended laboratory culture and antimicrobial susceptibility testing timelines hinder rapid species identification and susceptibility profiling of bacterial pathogens associated with bovine respiratory disease, the most prevalent cause of cattle mortality in the United States. Whole-genome sequencing offers a culture-independent alternative to current bacterial identification methods, but requires a library of bacterial reference genomes for comparison. To contribute new bacterial genome assemblies and evaluate genetic diversity and variation in antimicrobial resistance genotypes, whole-genome sequencing was performed on bovine respiratory disease-associated bacterial isolates (Histophilus somni, Mycoplasma bovis, Mannheimia haemolytica, and Pasteurella multocida) from dairy and beef cattle. One hundred genomically distinct assemblies were added to the NCBI database, doubling the available genomic sequences for these four species. Computer-based methods identified 11 predicted antimicrobial resistance genes in three species, with none being detected in M. bovis While computer-based analysis can identify antibiotic resistance genes within whole-genome sequences (genotype), it may not predict the actual antimicrobial resistance observed in a living organism (phenotype). Antimicrobial susceptibility testing on 64 H. somni, M. haemolytica, and P. multocida isolates had an overall concordance rate between genotype and phenotypic resistance to the associated class of antimicrobials of 72.7% (P < 0.001), showing substantial discordance. Concordance rates varied greatly among different antimicrobial, antibiotic resistance gene, and bacterial species combinations. This suggests that antimicrobial susceptibility phenotypes are needed to complement genomically predicted antibiotic resistance gene genotypes to better understand how the presence of antibiotic resistance genes within a given bacterial species could potentially impact optimal bovine respiratory disease treatment and morbidity/mortality outcomes

Core Outcome Set-STAndards for Development: The COS-STAD recommendations

Background The use of core outcome sets (COS) ensures that researchers measure and report those outcomes that are most likely to be relevant to users of their research. Several hundred COS projects have been systematically identified to date, but there has been no formal quality assessment of these studies. The Core Outcome Set-STAndards for Development (COS-STAD) project aimed to identify minimum standards for the design of a COS study agreed upon by an international group, while other specific guidance exists for the final reporting of COS development studies (Core Outcome Set-STAndards for Reporting [COS-STAR]). Methods and findings An international group of experienced COS developers, methodologists, journal editors, potential users of COS (clinical trialists, systematic reviewers, and clinical guideline developers), and patient representatives produced the COS-STAD recommendations to help improve the quality of COS development and support the assessment of whether a COS had been developed using a reasonable approach. An open survey of experts generated an initial list of items, which was refined by a 2-round Delphi survey involving nearly 250 participants representing key stakeholder groups. Participants assigned importance ratings for each item using a 1–9 scale. Consensus that an item should be included in the set of minimum standards was defined as at least 70% of the voting participants from each stakeholder group providing a score between 7 and 9. The Delphi survey was followed by a consensus discussion with the study management group representing multiple stakeholder groups. COS-STAD contains 11 minimum standards that are the minimum design recommendations for all COS development projects. The recommendations focus on 3 key domains: the scope, the stakeholders, and the consensus process. Conclusions The COS-STAD project has established 11 minimum standards to be followed by COS developers when planning their projects and by users when deciding whether a COS has been developed using reasonable methods

Using 3-D mapping to understand an Upper Ordovician buildup and facies complex in the upper Lexington Limestone, central Kentucky, USA

The upper parts of the Upper Ordovician Lexington Limestone in central Kentucky, USA, are interpreted to reflect a structurally controlled carbonate buildup, represented by a facies mosaic of shoal complexes and interbedded shale units. Facies intertonguing is complex and two-dimensional (2-D) mapping has been difficult. In this project, we converted 2-D maps to 3-D maps to show the extent of various facies and the complex nature of intertonguing. The resulting 3-D maps can be viewed from various vantage points and show the likely influence of basement structures as well as the results of post-depositional structural activity

SRAdb: query and use public next-generation sequencing data from within R

Abstract Background The Sequence Read Archive (SRA) is the largest public repository of sequencing data from the next generation of sequencing platforms including Illumina (Genome Analyzer, HiSeq, MiSeq, .etc), Roche 454 GS System, Applied Biosystems SOLiD System, Helicos Heliscope, PacBio RS, and others. Results SRAdb is an attempt to make queries of the metadata associated with SRA submission, study, sample, experiment and run more robust and precise, and make access to sequencing data in the SRA easier. We have parsed all the SRA metadata into a SQLite database that is routinely updated and can be easily distributed. The SRAdb R/Bioconductor package then utilizes this SQLite database for querying and accessing metadata. Full text search functionality makes querying metadata very flexible and powerful. Fastq files associated with query results can be downloaded easily for local analysis. The package also includes an interface from R to a popular genome browser, the Integrated Genomics Viewer. Conclusions SRAdb Bioconductor package provides a convenient and integrated framework to query and access SRA metadata quickly and powerfully from within R.</p