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Sample records for humans preliminary analyses

  1. SARDA HITL Preliminary Human Factors Measures and Analyses

    NASA Technical Reports Server (NTRS)

    Hyashi, Miwa; Dulchinos, Victoria

    2012-01-01

    Human factors data collected during the SARDA HITL Simulation Experiment include a variety of subjective measures, including the NASA TLX, questionnaire questions regarding situational awareness, advisory usefulness, UI usability, and controller trust. Preliminary analysis of the TLX data indicate that workload may not be adversely affected by use of the advisories, additionally, the controller's subjective ratings of the advisories may suggest acceptance of the tool.

  2. Preliminary analyses of scenarios for potential human interference for repositories in three salt formations

    SciTech Connect

    Not Available

    1985-10-01

    Preliminary analyses of scenarios for human interference with the performance of a radioactive waste repository in a deep salt formation are presented. The following scenarios are analyzed: (1) the U-Tube Connection Scenario involving multiple connections between the repository and the overlying aquifer system; (2) the Single Borehole Intrusion Scenario involving penetration of the repository by an exploratory borehole that simultaneously connects the repository with overlying and underlying aquifers; and (3) the Pressure Release Scenario involving inflow of water to saturate any void space in the repository prior to creep closure with subsequent release under near lithostatic pressures following creep closure. The methodology to evaluate repository performance in these scenarios is described and this methodology is applied to reference systems in three candidate formations: bedded salt in the Palo Duro Basin, Texas; bedded salt in the Paradox Basin, Utah; and the Richton Salt Dome, Mississippi, of the Gulf Coast Salt Dome Basin.

  3. Stereological analyses of the whole human pancreas

    PubMed Central

    Poudel, Ananta; Fowler, Jonas L.; Zielinski, Mark C.; Kilimnik, German; Hara, Manami

    2016-01-01

    The large size of human tissues requires a practical stereological approach to perform a comprehensive analysis of the whole organ. We have developed a method to quantitatively analyze the whole human pancreas, as one of the challenging organs to study, in which endocrine cells form various sizes of islets that are scattered unevenly throughout the exocrine pancreas. Furthermore, the human pancreas possesses intrinsic characteristics of intra-individual variability, i.e. regional differences in endocrine cell/islet distribution, and marked inter-individual heterogeneity regardless of age, sex and disease conditions including obesity and diabetes. The method is built based on large-scale image capture, computer-assisted unbiased image analysis and quantification, and further mathematical analyses, using widely-used software such as Fiji/ImageJ and MATLAB. The present study includes detailed protocols of every procedure as well as all the custom-written computer scripts, which can be modified according to specific experimental plans and specimens of interest. PMID:27658965

  4. Preliminary Framework for Human-Automation Collaboration

    SciTech Connect

    Oxstrand, Johanna Helene; Le Blanc, Katya Lee; Spielman, Zachary Alexander

    2015-09-01

    The Department of Energy’s Advanced Reactor Technologies Program sponsors research, development and deployment activities through its Next Generation Nuclear Plant, Advanced Reactor Concepts, and Advanced Small Modular Reactor (aSMR) Programs to promote safety, technical, economical, and environmental advancements of innovative Generation IV nuclear energy technologies. The Human Automation Collaboration (HAC) Research Project is located under the aSMR Program, which identifies developing advanced instrumentation and controls and human-machine interfaces as one of four key research areas. It is expected that the new nuclear power plant designs will employ technology significantly more advanced than the analog systems in the existing reactor fleet as well as utilizing automation to a greater extent. Moving towards more advanced technology and more automation does not necessary imply more efficient and safer operation of the plant. Instead, a number of concerns about how these technologies will affect human performance and the overall safety of the plant need to be addressed. More specifically, it is important to investigate how the operator and the automation work as a team to ensure effective and safe plant operation, also known as the human-automation collaboration (HAC). The focus of the HAC research is to understand how various characteristics of automation (such as its reliability, processes, and modes) effect an operator’s use and awareness of plant conditions. In other words, the research team investigates how to best design the collaboration between the operators and the automated systems in a manner that has the greatest positive impact on overall plant performance and reliability. This report addresses the Department of Energy milestone M4AT-15IN2302054, Complete Preliminary Framework for Human-Automation Collaboration, by discussing the two phased development of a preliminary HAC framework. The framework developed in the first phase was used as the

  5. 76 FR 52945 - Chlorpyrifos Registration Review; Preliminary Human Health Risk Assessment; Extension of Comment...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-24

    ... AGENCY Chlorpyrifos Registration Review; Preliminary Human Health Risk Assessment; Extension of Comment... availability of the chlorpyrifos registration review; preliminary human health risk assessment. This document... for the chlorpyrifos reregistration review, preliminary human health risk assessment, established...

  6. Human-Robot Emergency Response - Experimental Platform and Preliminary Dataset

    DTIC Science & Technology

    2014-07-28

    Human -Robot Emergency Response - Experimental Platform and Preliminary Dataset Technical Report #UM-CS-2014-006 Hee-Tae Jung, Takeshi Takahashi,and...2014 Abstract This paper presents progress towards a research infrastructure for studying human -robot performance in laboratory emergency response...scenarios and a preliminary dataset. It incorporates an emergency response team that is composed of a human participant, n ≤ 4 vision sensors in a

  7. Updated Human Health Risk Analyses for Chlorpyrifos

    EPA Pesticide Factsheets

    EPA has revised the human health hazard assessment and drinking water exposure assessment for chlorpyrifos that supported our October 2015 proposal to revoke all food residue tolerances for chlorpyrifos.

  8. Kinematic and Moisture Environments of Convective Systems During TRMM-LBA: Preliminary Sounding Analyses

    NASA Technical Reports Server (NTRS)

    Halverson, J. B.; Rickenbach, T.; Pierce, H.; Roy, B.; Ferreira, R. N.; Fisch, G.; Einaudi, Franco (Technical Monitor)

    2000-01-01

    Preliminary radiosonde data are analyzed from a four station observation network that operated during TRMM-LBA. These data, which are undergoing quality control, are used to construct mean vertical profiles and time-height sections of u- and v- wind components, and also filtered time series analyses of layer mean relative humidity. Trends are identified in the humidity data which appear similar at all sites, and correlate well with multi-week changes in wind regime identified by Rickenbach et al. Higher-frequency modes of variation (3-5 day) also occur in the humidity and upper tropospheric winds and are spatially coherent among the four locations. The causes of these variations are explored, including interactions among upper tropospheric synoptic features. Finally, an attempt is made to relate the general morphology of convective systems to the vertical shear structure and thermodynamic changes that accompany contrasting wind regimes.

  9. Preliminary Results of Ancillary Safety Analyses Supporting TREAT LEU Conversion Activities

    SciTech Connect

    Brunett, A. J.; Fei, T.; Strons, P. S.; Papadias, D. D.; Hoffman, E. A.; Kontogeorgakos, D. C.; Connaway, H. M.; Wright, A. E.

    2015-10-01

    Report (FSAR) [3]. Depending on the availability of historical data derived from HEU TREAT operation, results calculated for the LEU core are compared to measurements obtained from HEU TREAT operation. While all analyses in this report are largely considered complete and have been reviewed for technical content, it is important to note that all topics will be revisited once the LEU design approaches its final stages of maturity. For most safety significant issues, it is expected that the analyses presented here will be bounding, but additional calculations will be performed as necessary to support safety analyses and safety documentation. It should also be noted that these analyses were completed as the LEU design evolved, and therefore utilized different LEU reference designs. Preliminary shielding, neutronic, and thermal hydraulic analyses have been completed and have generally demonstrated that the various LEU core designs will satisfy existing safety limits and standards also satisfied by the existing HEU core. These analyses include the assessment of the dose rate in the hodoscope room, near a loaded fuel transfer cask, above the fuel storage area, and near the HEPA filters. The potential change in the concentration of tramp uranium and change in neutron flux reaching instrumentation has also been assessed. Safety-significant thermal hydraulic items addressed in this report include thermally-induced mechanical distortion of the grid plate, and heating in the radial reflector.

  10. Human Health Effects, Task Force Assessment, Preliminary Report.

    ERIC Educational Resources Information Center

    Aronow, Wilbert S.; And Others

    Presented in this preliminary report is one of seven assessments conducted by a special task force of Project Clean Air, the Human Health Effects Task Force. The reports summarize assessments of the state of knowledge on various air pollution problems, particularly in California, and make tentative recommendations as to what the University of…

  11. A Preliminary Classification of Human Functional Sexual Disorders

    ERIC Educational Resources Information Center

    Sharpe, Lawrence; And Others

    1976-01-01

    A preliminary classification is presented for functional human sexual disorders. This system is based on objective behavior and reports of distress. Five categories of sexual disorders are proposed, including the behavioral, psychological and informational components of sexual functioning in the individual and the couple. (Author)

  12. Template for Performing Human Reliability Analyses, Lesson Plans

    SciTech Connect

    Hunnaman, W., I. B. Wall

    2002-06-30

    Probabilistic Safety Analyses incorporate Human Reliability Assessment (HRA) to account for possible errors by a nuclear power plant operating crew both prior to and during postulated accidents. Studies have shown that human errors are large contributors to the likelihood of such accidents. A cadre of experts has developed HRA technology by applying it in many risk studies. The report provides insights to non-experts for application of HRA concepts in to specific nuclear plant PSAs.

  13. Human excimer laser corneal surgery: preliminary report.

    PubMed Central

    L'Esperance, F A; Taylor, D M; Del Pero, R A; Roberts, A; Gigstad, J; Stokes, M T; Warner, J W; Telfair, W B; Martin, C A; Yoder, P R

    1988-01-01

    The first human trial utilizing the argon fluoride excimer laser at 193 nm to produce a superficial keratectomy in ten human eyes has been described with the histopathological evaluation of four eyes and the longer gross appearance of six eyes at intervals extending to 10 months post-excimer laser treatment. The process of laser superficial keratectomy has proved to be one of the promising areas of surgical intervention for reconstructive or refractive keratoplasty in the future. Intensive investigations need to be undertaken on the corneal wound healing process following laser ablation as well as the nature, and long-term stability of the corneal excisions or induced refractive corrections. It is essential that the optimal laser parameters be established for the various refractive corrections and other corneal surgical techniques, and that pathophysiologic and histopathologic changes that have been induced by the excimer laser-corneal tissue interaction in animals and humans be critically and extensively analyzed. Images FIGURE 1 FIGURE 19 A FIGURE 19 B FIGURE 20 A FIGURE 20 B FIGURE 21 A FIGURE 21 B FIGURE 22 A FIGURE 22 B FIGURE 23 FIGURE 24 FIGURE 25 FIGURE 26 FIGURE 27 FIGURE 28 FIGURE 29 A FIGURE 29 B FIGURE 29 C FIGURE 29 D FIGURE 30 A FIGURE 30 B FIGURE 31 A FIGURE 31 B FIGURE 32 FIGURE 33 FIGURE 34 FIGURE 35 FIGURE 36 FIGURE 37 A FIGURE 37 B FIGURE 37 C FIGURE 38 A FIGURE 38 B FIGURE 39 A FIGURE 39 B FIGURE 39 C FIGURE 40 A FIGURE 40 B PMID:2979049

  14. Multitemporal satellite data analyses for archaeological mark detection: preliminary results in Italy and Argentina

    NASA Astrophysics Data System (ADS)

    Lasaponara, Rosa; Masini, Nicola

    2014-05-01

    within Basilicata and Puglia Region, southern Patagonia and Payunia-Campo Volcanicos Liancanelo e PayunMatru respectively, in Italy and Argentina. We focused our attention on diverse surfaces and soil types in different periods of the year in order to assess the capabilities of both optical and radar data to detect archaeological marks in different ecosystems and seasons. We investigated not only crop culture during the "favourable vegetative period" to enhance the presence of subsurface remains but also the "spectral response" of spontaneous, sparse herbaceous covers during periods considered and expected to be less favourable (as for example summer and winter) for this type of investigation. The main interesting results were the capability of radar (cosmoskymed) and multispectral optical data satellite data (Pleiades, Quickbird, Geoeye) to highlight the presence of structures below the surface even (i) in during period of years generally considered not "suitable for crop mark investigations" and even (ii) in areas only covered by sparse, spontaneous herbaceous plants in several test sites investigate din both Argentine and Italian areas of interest. Preliminary results conducted in both Italian and Argentina sites pointed out that Earth Observation (EO) technology can be successfully used for extracting useful information on traces the past human activities still fossilized in the modern landscape in different ecosystems and seasons. Moreover the multitemporal analyses of satellite data can fruitfully applied to: (i) improve knowledge, (ii) support monitoring of natural and cultural site, (iii) assess natural and man-made risks including emerging threats to the heritage sites. References Lasaponara R, N Masini 2009 Full-waveform Airborne Laser Scanning for the detection of medieval archaeological microtopographic relief Journal of Cultural Heritage 10, e78-e82 Ciminale M, D Gallo, R Lasaponara, N Masini 2009 A multiscale approach for reconstructing archaeological

  15. 9.4T Human MRI: Preliminary Results

    PubMed Central

    Vaughan, Thomas; DelaBarre, Lance; Snyder, Carl; Tian, Jinfeng; Akgun, Can; Shrivastava, Devashish; Liu, Wanzahn; Olson, Chris; Adriany, Gregor; Strupp, John; Andersen, Peter; Gopinath, Anand; van de Moortele, Pierre-Francois; Garwood, Michael; Ugurbil, Kamil

    2014-01-01

    This work reports the preliminary results of the first human images at the new high-field benchmark of 9.4T. A 65-cm-diameter bore magnet was used together with an asymmetric 40-cm-diameter head gradient and shim set. A multichannel transmission line (transverse electromagnetic (TEM)) head coil was driven by a programmable parallel transceiver to control the relative phase and magnitude of each channel independently. These new RF field control methods facilitated compensation for RF artifacts attributed to destructive interference patterns, in order to achieve homogeneous 9.4T head images or localize anatomic targets. Prior to FDA investigational device exemptions (IDEs) and internal review board (IRB)-approved human studies, preliminary RF safety studies were performed on porcine models. These data are reported together with exit interview results from the first 44 human volunteers. Although several points for improvement are discussed, the preliminary results demonstrate the feasibility of safe and successful human imaging at 9.4T. PMID:17075852

  16. Cartography of human diaphragmatic innervation: preliminary data.

    PubMed

    Verin, Eric; Marie, Jean-Paul; Similowski, Thomas

    2011-04-30

    In humans, anatomy indicates that the phrenic nerve mainly arises from the C4 cervical root, with variable C3 and C5 contributions. How this translates into functional innervation is unknown. The diaphragm response to electrical stimulation of C3, C4 and C5 was described in three patients undergoing surgical laryngeal reinnervation with an upper phrenic root (surface chest electrodes at anterior, lateral and posterior sites; oesophageal and gastric pressures (Pes and Pga) to derive transdiaphragmatic pressure (Pdi)). Anatomically, the phrenic nerve predominantly originated from C4. Phrenic stimulation elicited motor responses at the three sites in the three patients, as did C4 stimulation. It produced Pdi values of 9, 11, and 14cmH(2)O in the three patients, respectively, vs. 9, 9, and 7cmH(2)O for C4. C3 stimulation produced modest Pdi responses, whereas C5 stimulation could produce Pdi responses close to those observed with C4 stimulation. These singular observations confirm the dominance of C4 in diaphragm innervation but suggest than C5 can be of importance.

  17. Preliminary results of receiver function analyses at three sites across the Bransfield Strait, Antarctica

    NASA Astrophysics Data System (ADS)

    Martínez Arévalo, C.; Mancilla, F.; Almendros, J.; Aznarte, J. L.; Alguacil, G.

    2009-04-01

    In February 2008, in the framework of the International Polar Year 2008-2009 under grant POL2006-08663 of the spanish Ministry of Education, we deployed three broadband seismometers at points forming a N-S profile across the Bransfield Strait, Antarctica, in the region between the South Shetland Islands and the Antarctic Peninsula. This region is very interesting from a tectonic point of view, due to the opening of the rift of the Bransfield and the presence of the South Shetland microplate. For logistic reasons, our instruments where located in the vicinity of the Antarctic bases Juan Carlos I (Livingston Island), Gabriel de Castilla (Deception Island), and Primavera (Antarctic Peninsula). Each seismic station consisted of a broad-band, three-component electrolitic seismometer equiped with a 24-bit data acquisition system. The energy was provided by wind generators and solar panels connected to a battery bank, a combination that has been able to provide enough power at all sites, even during the Antarctic winter. All components were designed to function under the extreme conditions of the Antarctic weather. The main objective of this experiment was to use receiver function techniques on teleseism data to investigate the structure of the crust under the sites, in order to compare with other studies and shed light on the structure and tectonics of the region. During this past year, we have recorded several tens of teleseisms at distances appropriate for receiver function analyses. Preliminary results will add information on critical issues regarding the structure of the Bransfield Strait region. Although the Earth models obtained will contain extra information, we are specially interested in the determination of the depth of the Moho at each site, a controversial point in this area. These results will be compared with estimates obtained by different techniques. We are also interested in the determination of the vertical extent of the magma chamber recently imaged by

  18. HCV synthesis project: preliminary analyses of HCV prevalence in relation to age and duration of injection.

    PubMed

    Hagan, Holly; Des Jarlais, Don C; Stern, Rebecca; Lelutiu-Weinberger, Corina; Scheinmann, Roberta; Strauss, Shiela; Flom, Peter L

    2007-10-01

    Early acquisition of hepatitis C virus (HCV) infection appears to affect a substantial proportion of injection drug users (IDUs)--between 20 percent and 90 percent. Analysing the range of HCV prevalence estimates in new injectors may help identify factors that can be modified to reduce HCV transmission. The HCV Synthesis Project is a meta-analysis of studies of HCV epidemiology and prevention in drug users worldwide. In this preliminary analysis, we examined data from 127 studies of IDUs that reported HCV prevalence in relation to age or year since onset of drug injection, analysing heterogeneity and calculating summary statistics where appropriate. Six studies reported gender-specific HCV prevalence rates among young or new injectors; the group mean prevalence was 47 percent for men and 44 percent for women (NS). Group mean age for HCV-negatives was 24.7 years (range 24-28) and 26.1 years (range 21-31) for HCV-positives (n=8 studies). Data were examined from 13 studies that compared HCV prevalence among young injectors to older injectors using 5-year age categories; substantial variation was present within these categories such that measures of central tendency were not calculated. Similarly, among studies reporting HCV prevalence among IDUs in relation to 1-year intervals of duration of injection (<1 year, <2 years, and <3 years), considerable variability was observed. Notably, there were studies in each category that reported prevalence of 70 percent or higher among recent-onset drug injectors. Our findings confirm previous studies reporting high risk of acquiring HCV shortly after onset of injection; thus, HCV prevention programmes must emphasize methods to reach new injectors. Future research should (1) report data on time to infection in depth, (2) provide detailed information on study methodology, and (3) characterize the research setting with respect to underlying factors that affect injection practices and networks. This will permit synthesis of a greater

  19. An Illumination Modeling System for Human Factors Analyses

    NASA Technical Reports Server (NTRS)

    Huynh, Thong; Maida, James C.; Bond, Robert L. (Technical Monitor)

    2002-01-01

    Seeing is critical to human performance. Lighting is critical for seeing. Therefore, lighting is critical to human performance. This is common sense, and here on earth, it is easily taken for granted. However, on orbit, because the sun will rise or set every 45 minutes on average, humans working in space must cope with extremely dynamic lighting conditions. Contrast conditions of harsh shadowing and glare is also severe. The prediction of lighting conditions for critical operations is essential. Crew training can factor lighting into the lesson plans when necessary. Mission planners can determine whether low-light video cameras are required or whether additional luminaires need to be flown. The optimization of the quantity and quality of light is needed because of the effects on crew safety, on electrical power and on equipment maintainability. To address all of these issues, an illumination modeling system has been developed by the Graphics Research and Analyses Facility (GRAF) and Lighting Environment Test Facility (LETF) in the Space Human Factors Laboratory at NASA Johnson Space Center. The system uses physically based ray tracing software (Radiance) developed at Lawrence Berkeley Laboratories, a human factors oriented geometric modeling system (PLAID) and an extensive database of humans and environments. Material reflectivity properties of major surfaces and critical surfaces are measured using a gonio-reflectometer. Luminaires (lights) are measured for beam spread distribution, color and intensity. Video camera performances are measured for color and light sensitivity. 3D geometric models of humans and the environment are combined with the material and light models to form a system capable of predicting lighting conditions and visibility conditions in space.

  20. Uncertainty quantification for personalized analyses of human proximal femurs.

    PubMed

    Wille, Hagen; Ruess, Martin; Rank, Ernst; Yosibash, Zohar

    2016-02-29

    Computational models for the personalized analysis of human femurs contain uncertainties in bone material properties and loads, which affect the simulation results. To quantify the influence we developed a probabilistic framework based on polynomial chaos (PC) that propagates stochastic input variables through any computational model. We considered a stochastic E-ρ relationship and a stochastic hip contact force, representing realistic variability of experimental data. Their influence on the prediction of principal strains (ϵ1 and ϵ3) was quantified for one human proximal femur, including sensitivity and reliability analysis. Large variabilities in the principal strain predictions were found in the cortical shell of the femoral neck, with coefficients of variation of ≈40%. Between 60 and 80% of the variance in ϵ1 and ϵ3 are attributable to the uncertainty in the E-ρ relationship, while ≈10% are caused by the load magnitude and 5-30% by the load direction. Principal strain directions were unaffected by material and loading uncertainties. The antero-superior and medial inferior sides of the neck exhibited the largest probabilities for tensile and compression failure, however all were very small (pf<0.001). In summary, uncertainty quantification with PC has been demonstrated to efficiently and accurately describe the influence of very different stochastic inputs, which increases the credibility and explanatory power of personalized analyses of human proximal femurs.

  1. Crystallization and preliminary crystallographic analysis of recombinant human galectin-1

    SciTech Connect

    Scott, Stacy A.; Scott, Ken; Blanchard, Helen

    2007-11-01

    Human galectin-1 has been cloned, expressed in E. coli, purified and crystallized in the presence of both lactose (ligand) and β-mercaptoethanol under six different conditions. The X-ray diffraction data obtained have enabled the assignment of unit-cell parameters for two novel crystal forms of human galectin-1. Galectin-1 is considered to be a regulator protein as it is ubiquitously expressed throughout the adult body and is responsible for a broad range of cellular regulatory functions. Interest in galectin-1 from a drug-design perspective is founded on evidence of its overexpression by many cancers and its immunomodulatory properties. The development of galectin-1-specific inhibitors is a rational approach to the fight against cancer because although galectin-1 induces a plethora of effects, null mice appear normal. X-ray crystallographic structure determination will aid the structure-based design of galectin-1 inhibitors. Here, the crystallization and preliminary diffraction analysis of human galectin-1 crystals generated under six different conditions is reported. X-ray diffraction data enabled the assignment of unit-cell parameters for crystals grown under two conditions, one belongs to a tetragonal crystal system and the other was determined as monoclinic P2{sub 1}, representing two new crystal forms of human galectin-1.

  2. Analyses of volatile organic compounds from human skin

    PubMed Central

    Gallagher, M.; Wysocki, C.J.; Leyden, J.J.; Spielman, A.I.; Sun, X.; Preti, G.

    2008-01-01

    Summary Background Human skin emits a variety of volatile metabolites, many of them odorous. Much previous work has focused upon chemical structure and biogenesis of metabolites produced in the axillae (underarms), which are a primary source of human body odour. Nonaxillary skin also harbours volatile metabolites, possibly with different biological origins than axillary odorants. Objectives To take inventory of the volatile organic compounds (VOCs) from the upper back and forearm skin, and assess their relative quantitative variation across 25 healthy subjects. Methods Two complementary sampling techniques were used to obtain comprehensive VOC profiles, viz., solid-phase micro extraction and solvent extraction. Analyses were performed using both gas chromatography/mass spectrometry and gas chromatography with flame photometric detection. Results Nearly 100 compounds were identified, some of which varied with age. The VOC profiles of the upper back and forearm within a subject were, for the most part, similar, although there were notable differences. Conclusions The natural variation in nonaxillary skin odorants described in this study provides a baseline of compounds we have identified from both endogenous and exogenous sources. Although complex, the profiles of volatile constituents suggest that the two body locations share a considerable number of compounds, but both quantitative and qualitative differences are present. In addition, quantitative changes due to ageing are also present. These data may provide future investigators of skin VOCs with a baseline against which any abnormalities can be viewed in searching for biomarkers of skin diseases. PMID:18637798

  3. Preliminary organic analyses of the DSDP /JOIDES/ cores - Legs V-IX.

    NASA Technical Reports Server (NTRS)

    Simoneit, B. R.; Burlingame, A. L.

    1972-01-01

    Descriptions of the methods used and results obtained in analyses of deep sea drilling cores. The analyses were performed in two phases (differing in degree of particularization) depending on the amount of core sample available. The results are presented in relation to the ages and to the fossil fauna and flora of the sediments.

  4. Low Order Modeling Tools for Preliminary Pressure Gain Combustion Benefits Analyses

    NASA Technical Reports Server (NTRS)

    Paxson, Daniel E.

    2012-01-01

    Pressure gain combustion (PGC) offers the promise of higher thermodynamic cycle efficiency and greater specific power in propulsion and power systems. This presentation describes a model, developed under a cooperative agreement between NASA and AFRL, for preliminarily assessing the performance enhancement and preliminary size requirements of PGC components either as stand-alone thrust producers or coupled with surrounding turbomachinery. The model is implemented in the Numerical Propulsion Simulation System (NPSS) environment allowing various configurations to be examined at numerous operating points. The validated model is simple, yet physics-based. It executes quickly in NPSS, yet produces realistic results.

  5. Crystallization and preliminary crystallographic analysis of recombinant human galectin-1

    PubMed Central

    Scott, Stacy A.; Scott, Ken; Blanchard, Helen

    2007-01-01

    Galectin-1 is considered to be a regulator protein as it is ubiquitously expressed throughout the adult body and is responsible for a broad range of cellular regulatory functions. Interest in galectin-1 from a drug-design perspective is founded on evidence of its overexpression by many cancers and its immunomodulatory properties. The development of galectin-1-specific inhibitors is a rational approach to the fight against cancer because although galectin-1 induces a plethora of effects, null mice appear normal. X-ray crystallographic structure determination will aid the structure-based design of galectin-1 inhibitors. Here, the crystallization and preliminary diffraction analysis of human galectin-1 crystals generated under six different conditions is reported. X-ray diffraction data enabled the assignment of unit-cell parameters for crystals grown under two conditions, one belongs to a tetragonal crystal system and the other was determined as monoclinic P21, representing two new crystal forms of human galectin-1. PMID:18007053

  6. On protection of Freedom's solar dynamic radiator from the orbital debris environment. Part 1: Preliminary analyses and testing

    NASA Technical Reports Server (NTRS)

    Rhatigan, Jennifer L.; Christiansen, Eric L.; Fleming, Michael L.

    1990-01-01

    A great deal of experimentation and analysis was performed to quantify penetration thresholds of components which will experience orbital debris impacts. Penetration was found to depend upon mission specific parameters such as orbital altitude, inclination, and orientation of the component; and upon component specific parameters such as material, density and the geometry particular to its shielding. Experimental results are highly dependent upon shield configuration and cannot be extrapolated with confidence to alternate shield configurations. Also, current experimental capabilities are limited to velocities which only approach the lower limit of predicted orbital debris velocities. Therefore, prediction of the penetrating particle size for a particular component having a complex geometry remains highly uncertain. An approach is described which was developed to assess on-orbit survivability of the solar dynamic radiator due to micrometeoroid and space debris impacts. Preliminary analyses are presented to quantify the solar dynamic radiator survivability, and include the type of particle and particle population expected to defeat the radiator bumpering (i.e., penetrate a fluid flow tube). Results of preliminary hypervelocity impact testing performed on radiator panel samples (in the 6 to 7 km/sec velocity range) are also presented. Plans for further analyses and testing are discussed. These efforts are expected to lead to a radiator design which will perform to requirements over the expected lifetime.

  7. On protection of Freedom's solar dynamic radiator from the orbital debris environment. Part 1. Preliminary analyses and testing

    SciTech Connect

    Rhatigan, J.L.; Christiansen, E.L.; Fleming, M.L.

    1990-01-01

    A great deal of experimentation and analysis was performed to quantify penetration thresholds of components which will experience orbital debris impacts. Penetration was found to depend upon mission specific parameters such as orbital altitude, inclination, and orientation of the component; and upon component specific parameters such as material, density and the geometry particular to its shielding. Experimental results are highly dependent upon shield configuration and cannot be extrapolated with confidence to alternate shield configurations. Also, current experimental capabilities are limited to velocities which only approach the lower limit of predicted orbital debris velocities. Therefore, prediction of the penetrating particle size for a particular component having a complex geometry remains highly uncertain. An approach is described which was developed to assess on-orbit survivability of the solar dynamic radiator due to micrometeoroid and space debris impacts. Preliminary analyses are presented to quantify the solar dynamic radiator survivability, and include the type of particle and particle population expected to defeat the radiator bumpering (i.e., penetrate a fluid flow tube). Results of preliminary hypervelocity impact testing performed on radiator panel samples (in the 6 to 7 km/sec velocity range) are also presented. Plans for further analyses and testing are discussed. These efforts are expected to lead to a radiator design which will perform to requirements over the expected lifetime.

  8. Operator Performance on Two Office Data Entry System Testbeds: Preliminary Analyses.

    DTIC Science & Technology

    1982-02-01

    er Navy Personnel Research and Development Center San Diego, California 92152 UNCLASSIFIED SICu111Tv CLASIFICATION OF T16 PAGE 1bm 0-" 8100-40...and Usability of Automated Personnel Information Systems. The objective of this subproject is to resolve fundamental human engineering design issues...R. C. Metric for evaluation of human -computer interaction in a personnel records task. In IEE Proceedings of the International Conference on

  9. Preliminary analyses for perchlorate in selected natural materials and their derivative products

    USGS Publications Warehouse

    Orris, G.J.; Harvey, G.J.; Tsui, D.T.; Eldrige, J.E.

    2003-01-01

    Increasing concern about sources of perchlorate contamination in ground and surface waters has led to interest in identifying potential sources of natural perchlorate and products derived from these natural sources. To date, most perchlorate found in ground and surface waters has been attributed to its major uses as an oxidizer in solid propellants for rockets, in fireworks and other explosives, and a variety of other uses of man-made perchlorate salts. However, perchlorate found in the soils, surface water, and ground water of some locations cannot be linked to an anthropogenic source. This paper contains preliminary data on the detection and non-detection of perchlorate in a variety of natural materials and their products, including some fertilizer materials. These data were previously presented at two conferences; once in poster session and once orally (Harvey and others, 1999; Orris and others, 2000). Although the results presented here are included in a journal article awaiting publication, the lack of public information on this topic has led to repeated requests for the data used as the basis for our presentations in 1999 and 2000.

  10. Lists and analyses of the mineral springs of the United States: A preliminary study

    USGS Publications Warehouse

    Peale, Albert C.

    1886-01-01

    In attempting the collection of data for the statement of the commercial value of the mineral waters of the country for publication in the report on the Mineral Resources of the United States, 1883 and 1884, it was necessary as a prerequisite to have a list of the springs from which these waters are derived. An examination of the few general works on the subject very soon showed that all existing lists were incomplete. The tables given in this paper were therefore compiled, as the first step in the preparation of the mineral spring statistics of the. United States, They were omitted from the paper published in Mr. Williams's report, for want of space. Since the appearance of that report they have been revised and, with the addition of such analyses as could be obtained, prepared for publication as a bulletin of the Survey.

  11. Preliminary Performance Analyses of the Constellation Program ARES 1 Crew Launch Vehicle

    NASA Technical Reports Server (NTRS)

    Phillips, Mark; Hanson, John; Shmitt, Terri; Dukemand, Greg; Hays, Jim; Hill, Ashley; Garcia, Jessica

    2007-01-01

    By the time NASA's Exploration Systems Architecture Study (ESAS) report had been released to the public in December 2005, engineers at NASA's Marshall Space Flight Center had already initiated the first of a series of detailed design analysis cycles (DACs) for the Constellation Program Crew Launch Vehicle (CLV), which has been given the name Ares I. As a major component of the Constellation Architecture, the CLV's initial role will be to deliver crew and cargo aboard the newly conceived Crew Exploration Vehicle (CEV) to a staging orbit for eventual rendezvous with the International Space Station (ISS). However, the long-term goal and design focus of the CLV will be to provide launch services for a crewed CEV in support of lunar exploration missions. Key to the success of the CLV design effort and an integral part of each DAC is a detailed performance analysis tailored to assess nominal and dispersed performance of the vehicle, to determine performance sensitivities, and to generate design-driving dispersed trajectories. Results of these analyses provide valuable design information to the program for the current design as well as provide feedback to engineers on how to adjust the current design in order to maintain program goals. This paper presents a condensed subset of the CLV performance analyses performed during the CLV DAC-1 cycle. Deterministic studies include development of the CLV DAC-1 reference trajectories, identification of vehicle stage impact footprints, an assessment of launch window impacts to payload performance, and the computation of select CLV payload partials. Dispersion studies include definition of input uncertainties, Monte Carlo analysis of trajectory performance parameters based on input dispersions, assessment of CLV flight performance reserve (FPR), assessment of orbital insertion accuracy, and an assessment of bending load indicators due to dispersions in vehicle angle of attack and side slip angle. A short discussion of the various

  12. Stardust Interstellar Preliminary Examination V: XRF analyses of interstellar dust candidates at ESRF ID13

    NASA Astrophysics Data System (ADS)

    Brenker, Frank E.; Westphal, Andrew J.; Vincze, Laszlo; Burghammer, Manfred; Schmitz, Sylvia; Schoonjans, Tom; Silversmit, Geert; Vekemans, Bart; Allen, Carlton; Anderson, David; Ansari, Asna; Bajt, SašA.; Bastien, Ron K.; Bassim, Nabil; Bechtel, Hans A.; Borg, Janet; Bridges, John; Brownlee, Donald E.; Burchell, Mark; Butterworth, Anna L.; Changela, Hitesh; Cloetens, Peter; Davis, Andrew M.; Doll, Ryan; Floss, Christine; Flynn, George; Fougeray, Patrick; Frank, David R.; Gainsforth, Zack; Grün, Eberhard; Heck, Philipp R.; Hillier, Jon K.; Hoppe, Peter; Hudson, Bruce; Huth, Joachim; Hvide, Brit; Kearsley, Anton; King, Ashley J.; Lai, Barry; Leitner, Jan; Lemelle, Laurence; Leroux, Hugues; Leonard, Ariel; Lettieri, Robert; Marchant, William; Nittler, Larry R.; Ogliore, Ryan; Ong, Wei Ja; Postberg, Frank; Price, Mark C.; Sandford, Scott A.; Tresseras, Juan-Angel Sans; Simionovici, Alexandre S.; Solé, Vicente A.; Srama, Ralf; Stadermann, Frank; Stephan, Thomas; Sterken, Veerle J.; Stodolna, Julien; Stroud, Rhonda M.; Sutton, Steven; Trieloff, Mario; Tsou, Peter; Tsuchiyama, Akira; Tyliszczak, Tolek; Korff, Joshua; Wordsworth, Naomi; Zevin, Daniel; Zolensky, Michael E.

    2014-09-01

    Here, we report analyses by synchrotron X-ray fluorescence microscopy of the elemental composition of eight candidate impact features extracted from the Stardust Interstellar Dust Collector (SIDC). Six of the features were unambiguous tracks, and two were crater-like features. Five of the tracks are so-called "midnight" tracks—that is, they had trajectories consistent with an origin either in the interstellar dust stream or as secondaries from impacts on the Sample Return Capsule (SRC). In a companion paper reporting synchrotron X-ray diffraction analyses of ISPE candidates, we show that two of these particles contain natural crystalline materials: the terminal particle of track 30 contains olivine and spinel, and the terminal particle of track 34 contains olivine. Here, we show that the terminal particle of track 30, Orion, shows elemental abundances, normalized to Fe, that are close to CI values, and a complex, fine-grained structure. The terminal particle of track 34, Hylabrook, shows abundances that deviate strongly from CI, but shows little fine structure and is nearly homogenous. The terminal particles of other midnight tracks, 29 and 37, had heavy element abundances below detection threshold. A third, track 28, showed a composition inconsistent with an extraterrestrial origin, but also inconsistent with known spacecraft materials. A sixth track, with a trajectory consistent with secondary ejecta from an impact on one of the spacecraft solar panels, contains abundant Ce and Zn. This is consistent with the known composition of the glass covering the solar panel. Neither crater-like feature is likely to be associated with extraterrestrial materials. We also analyzed blank aerogel samples to characterize background and variability between aerogel tiles. We found significant differences in contamination levels and compositions, emphasizing the need for local background subtraction for accurate quantification.

  13. Stardust Interstellar Preliminary Examination V: XRF Analyses of Interstellar Dust Candidates at ESRF ID13

    NASA Technical Reports Server (NTRS)

    Brenker, Frank E.; Westphal, Andrew J.; Simionovici, Alexandre S.; Flynn, George J.; Gainsforth, Zack; Allen, Carlton C.; Sanford, Scott; Zolensky, Michael E.; Bastien, Ron K.; Frank, David R.

    2014-01-01

    Here, we report analyses by synchrotron X-ray fluorescence microscopy of the elemental composition of eight candidate impact features extracted from the Stardust Interstellar Dust Collector (SIDC). Six of the features were unambiguous tracks, and two were crater-like features. Five of the tracks are so-called midnight tracks that is, they had trajectories consistent with an origin either in the interstellar dust stream or as secondaries from impacts on the Sample Return Capsule (SRC). In a companion paper reporting synchrotron X-ray diffraction analyses of ISPE candidates, we show that two of these particles contain natural crystalline materials: the terminal particle of track 30contains olivine and spinel, and the terminal particle of track 34 contains olivine. Here, we show that the terminal particle of track 30, Orion, shows elemental abundances, normalized to Fe, that are close to CI values, and a complex, fine-grained structure. The terminal particle of track 34, Hylabrook, shows abundances that deviate strongly from CI, but shows little fine structure and is nearly homogenous. The terminal particles of other midnight tracks, 29 and 37, had heavy element abundances below detection threshold. A third, track28, showed a composition inconsistent with an extraterrestrial origin, but also inconsistent with known spacecraft materials. A sixth track, with a trajectory consistent with secondary ejecta from an impact on one of the spacecraft solar panels, contains abundant Ce and Zn. This is consistent with the known composition of the glass covering the solar panel. Neither crater-like feature is likely to be associated with extraterrestrial materials. We also analyzed blank aerogel samples to characterize background and variability between aerogel tiles. We found significant differences in contamination levels and compositions, emphasizing the need for local background subtraction for accurate quantification.

  14. Preliminary analyses of WL experiment No. 701, space environment effects on operating fiber optic systems

    NASA Technical Reports Server (NTRS)

    Taylor, E. W.; Berry, J. N.; Sanchez, A. D.; Padden, R. J.; Chapman, S. P.

    1992-01-01

    A brief overview of the analyses performed to date on WL Experiment-701 is presented. Four active digital fiber optic links were directly exposed to the space environment for a period of 2114 days. The links were situated aboard the Long Duration Exposure Facility (LDEF) with the cabled, single fiber windings atop an experimental tray containing instrumentation for exercising the experiment in orbit. Despite the unplanned and prolonged exposure to trapped and galactic radiation, wide temperature extremes, atomic oxygen interactions, and micro-meteorite and debris impacts, in most instances the optical data links performed well within the experimental limits. Analysis of the recorded orbital data clearly indicates that fiber optic applications in space will meet with success. Ongoing tests and analysis of the experiment at the Phillips Laboratory's Optoelectronics Laboratory will expand this premise, and establish the first known and extensive database of active fiber optic link performance during prolonged space exposure. WL Exp-701 was designed as a feasibility demonstration for fiber optic technology in space applications, and to study the performance of operating fiber systems exposed to space environmental factors such as galactic radiation, and wide temperature cycling. WL Exp-701 is widely acknowledged as a benchmark accomplishment that clearly demonstrates, for the first time, that fiber optic technology can be successfully used in a variety of space applications.

  15. Preliminary analyses of space radiation protection for lunar base surface systems

    NASA Technical Reports Server (NTRS)

    Nealy, John E.; Wilson, John W.; Townsend, Lawrence W.

    1989-01-01

    Radiation shielding analyses are performed for candidate lunar base habitation modules. The study primarily addresses potential hazards due to contributions from the galactic cosmic rays. The NASA Langley Research Center's high energy nucleon and heavy ion transport codes are used to compute propagation of radiation through conventional and regolith shield materials. Computed values of linear energy transfer are converted to biological dose-equivalent using quality factors established by the International Commision of Radiological Protection. Special fluxes of heavy charged particles and corresponding dosimetric quantities are computed for a series of thicknesses in various shield media and are used as an input data base for algorithms pertaining to specific shielded geometries. Dosimetric results are presented as isodose contour maps of shielded configuration interiors. The dose predictions indicate that shielding requirements are substantial, and an abbreviated uncertainty analysis shows that better definition of the space radiation environment as well as improvement in nuclear interaction cross-section data can greatly increase the accuracy of shield requirement predictions.

  16. Compositional diversity of the lunar North Pole: Preliminary analyses of Galileo SSI data

    NASA Technical Reports Server (NTRS)

    Pieters, C. M.; Belton, M.; Head, James W.; Greeley, R.; Mcewen, A.; Fischer, E. M.; Sunshine, J. M.; Klaasen, K.; Plutchak, J.; Neukum, G.

    1993-01-01

    In December 1992, the Galileo spacecraft passed through the Earth-Moon system for its final gravity assist to Jupiter. The SSI camera obtained several 6-color mosaics of the lunar north polar region and the sunlit nearside and eastern limb at approximately 1.3 km/pixel. Initial analyses have concentrated on the north polar areas to assess the composition of the crust in that region. Representative 6-color calibration SSI spectra (typically 5 x 5 pixels) are shown. Photometric corrections have not yet been applied, and all spectra are scaled to unity at 0.56 microns. The data were first calibrated relative to MS2, a standard are in Mare Serenitatis (18.7 deg N, 21.5 deg E), and the top four plots of highlands, highland craters, maria, and mare craters are displayed relative to MS2. SSI spectra of areas measured with telescopic data (mare MT1/MS2 and mare crater MSA/MS2) agree well with previous data, confirming that the calibration procedures and SSI data are spectrally accurate. The bottom three plots of craters/sun have been calibrated to reflectance using previously obtained telescopic spectra of Apollo 16/MS2 (shown with Highlands/MS2) and laboratory spectra of mature Apollo 16 soil (shown for reference with the Highland Craters/Sun). Although some variations in these spectra mimic previously observed spectra of lunar terrains, several characteristics are unusual. Familiar and unfamiliar properties are observed in these northern latitudes and both types merit further investigation in their geologic context.

  17. Venomics analyses of the skin secretion of Dermatonotus muelleri: Preliminary proteomic and metabolomic profiling.

    PubMed

    Cavalcante, Ingrid Duarte; Antoniazzi, Marta Maria; Jared, Carlos; Pires, Osmindo R; Sciani, Juliana Mozer; Pimenta, Daniel Carvalho

    2017-05-01

    Dermatonotus muelleri is the sole species of the Dermatonotus genus and inhabits Argentina, Bolivia, Brazil and Paraguay. This animal exhibits an explosive reproductive behavior during the Southern spring months, which lasts only for five days. Moreover, this animal displays specific adaptations to the habitat resulting in the energy conservation needed during either the intense reproduction period or times of estivation. During dry seasons and/or food shortages D. muelleri can survive because its food specialization and ability to dig an underground chamber for protection. Few literature is available on this amphibian and no biochemical characterization has ever been performed on the animal's skin secretion. This work, on the other hand, presents for the first time a venomic analysis of the major components present in the skin secretion of this microhylid. The crude skin secretion was obtained my mechanical stimulation and was analyzed according to one major criterion: >10 kDa or <10 kDa. The high molecular mass fraction was subjected to typical gel-based proteomic processing whereas the low molecular mass fraction was analyzed by liquid chromatography, mass spectrometry and nuclear magnetic resonance (NMR), yielding an overall 'venomics' approach. No classical/evident toxin was detected, but peptidases (metallo and serino) and structural proteins could be identified. In the low molecular mass fraction no peptides were detected, as well as no typical alkaloid or steroid. On the other hand, the amino acid tryptophan could be identified and a typical sugar spectrum was obtained in the NMR analyses. Altogether these findings point out to the fact that D. muelleri skin secretion is unique and the molecular arsenal present herein is yet to be explored; therefore, this venomics study is only the beginning.

  18. New Careers in Human Service: A Challenge to the Two-Year College. A Preliminary Report.

    ERIC Educational Resources Information Center

    Burns, Martha A.

    This preliminary report, conducted over a 1-year period, involves the identification and study of educational programs that are preparing paraprofessional personnel for human service occupations. The objectives are to describe the present status of human service occupational education, to anticipate the direction human service education will take…

  19. Ecogeographical associations between climate and human body composition: analyses based on anthropometry and skinfolds.

    PubMed

    Wells, Jonathan C K

    2012-02-01

    In the 19th century, two "ecogeographical rules" were proposed hypothesizing associations of climate with mammalian body size and proportions. Data on human body weight and relative leg length support these rules; however, it is unknown whether such associations are attributable to lean tissue (the heat-producing component) or fat (energy stores). Data on weight, height, and two skinfold thickness were obtained from the literature for 137 nonindustrialized populations, providing 145 male and 115 female individual samples. A variety of indices of adiposity and lean mass were analyzed. Preliminary analyses indicated secular increases in skinfolds in men but not women, and associations of age and height with lean mass in both sexes. Decreasing annual temperature was associated with increasing body mass index (BMI), and increasing triceps but not subscapular skinfold. After adjusting for skinfolds, decreasing temperature remained associated with increasing BMI. These results indicate that colder environments favor both greater peripheral energy stores, and greater lean mass. Contrasting results for triceps and subscapular skinfolds might be due to adaptive strategies either constraining central adiposity in cold environments to reduce cardiovascular risk, or favoring central adiposity in warmer environments to maintain energetic support of the immune system. Polynesian populations were analyzed separately and contradicted all of the climate trends, indicating support for the hypothesis that they are cold-adapted despite occupying a tropical region. It is unclear whether such associations emerge through natural selection or through trans-generational and life-course plasticity. These findings nevertheless aid understanding of the wide variability in human physique and adiposity.

  20. Crucial role of detailed function, task, timeline, link and human vulnerability analyses in HRA

    SciTech Connect

    Ryan, T.G.; Haney, L.N.; Ostrom, L.T.

    1992-10-01

    This paper addresses one major cause for large uncertainties in human reliability analysis (HRA) results, that is, an absence of detailed function, task, timeline, link and human vulnerability analyses. All too often this crucial step in the HRA process is done in a cursory fashion using word of mouth or written procedures which themselves may incompletely or inaccurately represent the human action sequences and human error vulnerabilities being analyzed. The paper examines the potential contributions these detailed analyses can make in achieving quantitative and qualitative HRA results which are: (1) creditable, that is, minimize uncertainty, (2) auditable, that is, systematically linking quantitative results and qualitative information from which the results are derived, (3) capable of supporting root cause analyses on human reliability factors determined to be major contributors to risk, and (4) capable of repeated measures and being combined with similar results from other analyses to examine HRA issues transcending individual systems and facilities. Based on experience analyzing test and commercial nuclear reactors, and medical applications of nuclear technology, an iterative process is suggested for doing detailed function, task, timeline, link and human vulnerability analyses using documentation reviews, open-ended and structured interviews, direct observations, and group techniques. Finally, the paper concludes that detailed analyses done in this manner by knowledgeable human factors practitioners, can contribute significantly to the credibility, auditability, causal factor analysis, and combining goals of the HRA.

  1. Crucial role of detailed function, task, timeline, link and human vulnerability analyses in HRA. [Human Reliability Analysis (HRA)

    SciTech Connect

    Ryan, T.G.; Haney, L.N.; Ostrom, L.T.

    1992-01-01

    This paper addresses one major cause for large uncertainties in human reliability analysis (HRA) results, that is, an absence of detailed function, task, timeline, link and human vulnerability analyses. All too often this crucial step in the HRA process is done in a cursory fashion using word of mouth or written procedures which themselves may incompletely or inaccurately represent the human action sequences and human error vulnerabilities being analyzed. The paper examines the potential contributions these detailed analyses can make in achieving quantitative and qualitative HRA results which are: (1) creditable, that is, minimize uncertainty, (2) auditable, that is, systematically linking quantitative results and qualitative information from which the results are derived, (3) capable of supporting root cause analyses on human reliability factors determined to be major contributors to risk, and (4) capable of repeated measures and being combined with similar results from other analyses to examine HRA issues transcending individual systems and facilities. Based on experience analyzing test and commercial nuclear reactors, and medical applications of nuclear technology, an iterative process is suggested for doing detailed function, task, timeline, link and human vulnerability analyses using documentation reviews, open-ended and structured interviews, direct observations, and group techniques. Finally, the paper concludes that detailed analyses done in this manner by knowledgeable human factors practitioners, can contribute significantly to the credibility, auditability, causal factor analysis, and combining goals of the HRA.

  2. Overview of a Preliminary Destination Mission Concept for a Human Orbital Mission to the Martial Moons

    NASA Technical Reports Server (NTRS)

    Mazanek, D. D.; Abell, P. A.; Antol, J.; Barbee, B. W.; Beaty, D. W.; Bass, D. S.; Castillo-Rogez, J. C.; Coan, D. A.; Colaprete, A.; Daugherty, K. J.; Drake, B. G.; Earle, K. D.; Graham, L. D.; Hembree, R. M.; Hoffman, S. J.; Jefferies, S. A.; Lupisella, M. L.; Reeves, David M.

    2012-01-01

    The National Aeronautics and Space Administration s Human Spaceflight Architecture Team (HAT) has been developing a preliminary Destination Mission Concept (DMC) to assess how a human orbital mission to one or both of the Martian moons, Phobos and Deimos, might be conducted as a follow-on to a human mission to a near-Earth asteroid (NEA) and as a possible preliminary step prior to a human landing on Mars. The HAT Mars-Phobos-Deimos (MPD) mission also permits the teleoperation of robotic systems by the crew while in the Mars system. The DMC development activity provides an initial effort to identify the science and exploration objectives and investigate the capabilities and operations concepts required for a human orbital mission to the Mars system. In addition, the MPD Team identified potential synergistic opportunities via prior exploration of other destinations currently under consideration.

  3. Crystallization And Preliminary Crystallographic Analysis of Recombinant Human Galectin-1

    SciTech Connect

    Scott, S.A.; Scott, K.; Blanchard, H.

    2009-06-04

    Human galectin-1 has been cloned, expressed in E. coli, purified and crystallized in the presence of both lactose (ligand) and {beta}-mercaptoethanol under six different conditions. The X-ray diffraction data obtained have enabled the assignment of unit-cell parameters for two novel crystal forms of human galectin-1.

  4. Preliminary Work Domain Analysis for Human Extravehicular Activity

    NASA Technical Reports Server (NTRS)

    McGuire, Kerry; Miller, Matthew; Feigh, Karen

    2015-01-01

    A work domain analysis (WDA) of human extravehicular activity (EVA) is presented in this study. A formative methodology such as Cognitive Work Analysis (CWA) offers a new perspective to the knowledge gained from the past 50 years of living and working in space for the development of future EVA support systems. EVA is a vital component of human spaceflight and provides a case study example of applying a work domain analysis (WDA) to a complex sociotechnical system. The WDA presented here illustrates how the physical characteristics of the environment, hardware, and life support systems of the domain guide the potential avenues and functional needs of future EVA decision support system development.

  5. Somatic Variants in the Human Lens Epithelium: A Preliminary Assessment

    PubMed Central

    Mesa, Rosana; Tyagi, Manoj; Harocopos, George; Vollman, David; Bassnett, Steven

    2016-01-01

    Purpose We hypothesize that somatic mutations accumulate in cells of the human lens and may contribute to the development of cortical or posterior sub-capsular cataracts. Here, we used a Next-generation sequencing (NGS) strategy to screen for low-allelic frequency variants in DNA extracted from human lens epithelial samples. Methods Next-Generation sequencing of 151 cancer-related genes (WUCaMP2 panel) was performed on DNA extracted from post-mortem or surgical specimens obtained from 24 individuals. Usually, pairwise comparisons were made between two or more ocular samples from the same individual, allowing putative somatic variants detected in lens samples to be differentiated from germline variants. Results Use of a targeted hybridization approach enabled high sequence coverage (>1000-fold) of the WUCaMP2 genes. In addition to high-frequency variants (corresponding to homozygous or heterozygous SNPs and Indels), somatic variants with allelic frequencies of 1-4% were detected in the lens epithelial samples. The presence of one such variant, a T > C point substitution at position 32907082 in BRCA2, was verified subsequently using droplet digital PCR. Conclusions Low-allelic fraction variants are present in the human lens epithelium, at frequencies consistent with the presence of millimeter-sized clones. PMID:27537255

  6. Crystallization and preliminary crystallographic analysis of human glycosylated haemoglobin

    SciTech Connect

    Syakhovich, Vitaly E.; Saraswathi, N. T.; Ruff, Marc; Bokut, Sergey B.; Moras, Dino

    2006-02-01

    Non enzymatic modification of haemoglobin by glucose plays an important role in diabetes pathogenesis. Here the purification, characterization and crystallization of human glycosylated haemoglobin are reported. Human glycosylated haemoglobin A{sub 1C} is a stable minor variant formed in vivo by post-translational modification of the main form of haemoglobin by glucose. Crystals of oxyHbA{sub 1C} were obtained using the hanging-drop vapour-diffusion method and PEG as precipitant. The diffraction pattern of the crystal extends to a resolution of 2.3 Å at 120 K. The crystals belong to space group C2, with unit-cell parameters a = 237.98, b = 59.27, c = 137.02 Å, α = 90.00, β = 125.40, γ = 90.00°. The presence of two and a half molecules per asymmetric unit gives a crystal volume per protein weight (V{sub M}) of 9.70 Å{sup 3} Da{sup −1} and a solvent content of 49%.

  7. Preliminary Exploration of Adaptive State Predictor Based Human Operator Modeling

    NASA Technical Reports Server (NTRS)

    Trujillo, Anna C.; Gregory, Irene M.

    2012-01-01

    Control-theoretic modeling of the human operator dynamic behavior in manual control tasks has a long and rich history. In the last two decades, there has been a renewed interest in modeling the human operator. There has also been significant work on techniques used to identify the pilot model of a given structure. The purpose of this research is to attempt to go beyond pilot identification based on collected experimental data and to develop a predictor of pilot behavior. An experiment was conducted to quantify the effects of changing aircraft dynamics on an operator s ability to track a signal in order to eventually model a pilot adapting to changing aircraft dynamics. A gradient descent estimator and a least squares estimator with exponential forgetting used these data to predict pilot stick input. The results indicate that individual pilot characteristics and vehicle dynamics did not affect the accuracy of either estimator method to estimate pilot stick input. These methods also were able to predict pilot stick input during changing aircraft dynamics and they may have the capability to detect a change in a subject due to workload, engagement, etc., or the effects of changes in vehicle dynamics on the pilot.

  8. Analysis of organochlorine pesticides in human milk: preliminary results.

    PubMed

    Campoy, C; Jiménez, M; Olea-Serrano, M F; Moreno-Frías, M; Cañabate, F; Olea, N; Bayés, R; Molina-Font, J A

    2001-11-01

    In the face of evidence of human milk contamination by organochlorine pesticides, an analysis was performed on samples of milk obtained from healthy lactating women in the provinces of Granada and Almeria in Southern Spain. The samples were obtained by the Neonate Section of the Department of Pediatrics of Granada University Hospital (Neonatology Division) and by the Neonatal Service of Poniente Hospital in El Ejido, Almería. A liquid-liquid extraction procedure was performed. The cleaning of the sample before gas chromatography-mass spectrometry (GC-MS) used silica Sep-Pak. Among other pesticides, aldrin, dieldrin, DDT and its metabolites, lindane, methoxychlor and endosulfan were identified. The presence of these products was confirmed by mass spectrometry. The identification and quantification of these organochlorine molecules is important because they have estrogenic effects.

  9. Mitochondrial Genome Analyses Suggest Multiple Trichuris Species in Humans, Baboons, and Pigs from Different Geographical Regions

    PubMed Central

    Hawash, Mohamed B. F.; Andersen, Lee O.; Gasser, Robin B.; Stensvold, Christen Rune; Nejsum, Peter

    2015-01-01

    Background The whipworms Trichuris trichiura and Trichuris suis are two parasitic nematodes of humans and pigs, respectively. Although whipworms in human and non-human primates historically have been referred to as T. trichiura, recent reports suggest that several Trichuris spp. are found in primates. Methods and Findings We sequenced and annotated complete mitochondrial genomes of Trichuris recovered from a human in Uganda, an olive baboon in the US, a hamadryas baboon in Denmark, and two pigs from Denmark and Uganda. Comparative analyses using other published mitochondrial genomes of Trichuris recovered from a human and a porcine host in China and from a françois’ leaf-monkey (China) were performed, including phylogenetic analyses and pairwise genetic and amino acid distances. Genetic and protein distances between human Trichuris in Uganda and China were high (~19% and 15%, respectively) suggesting that they represented different species. Trichuris from the olive baboon in US was genetically related to human Trichuris in China, while the other from the hamadryas baboon in Denmark was nearly identical to human Trichuris from Uganda. Baboon-derived Trichuris was genetically distinct from Trichuris from françois’ leaf monkey, suggesting multiple whipworm species circulating among non-human primates. The genetic and protein distances between pig Trichuris from Denmark and other regions were roughly 9% and 6%, respectively, while Chinese and Ugandan whipworms were more closely related. Conclusion and Significance Our results indicate that Trichuris species infecting humans and pigs are phylogenetically distinct across geographical regions, which might have important implications for the implementation of suitable and effective control strategies in different regions. Moreover, we provide support for the hypothesis that Trichuris infecting primates represents a complex of cryptic species with some species being able to infect both humans and non-human primates

  10. GENE EXPRESSION IN THE TESTES OF NORMOSPERMIC VERSUS TERATOSPERMIC DOMESTIC CATS USING HUMAN CDNA MICROARRAY ANALYSES

    EPA Science Inventory

    GENE EXPRESSION IN THE TESTES OF NORMOSPERMIC VERSUS TERATOSPERMIC DOMESTIC CATS USING HUMAN cDNA MICROARRAY ANALYSES

    B.S. Pukazhenthi1, J. C. Rockett2, M. Ouyang3, D.J. Dix2, J.G. Howard1, P. Georgopoulos4, W.J. J. Welsh3 and D. E. Wildt1

    1Department of Reproductiv...

  11. Preliminary study to characterize plastic polymers using elemental analyser/isotope ratio mass spectrometry (EA/IRMS).

    PubMed

    Berto, Daniela; Rampazzo, Federico; Gion, Claudia; Noventa, Seta; Ronchi, Francesca; Traldi, Umberto; Giorgi, Giordano; Cicero, Anna Maria; Giovanardi, Otello

    2017-06-01

    Plastic waste is a growing global environmental problem, particularly in the marine ecosystems, in consideration of its persistence. The monitoring of the plastic waste has become a global issue, as reported by several surveillance guidelines proposed by Regional Sea Conventions (OSPAR, UNEP) and appointed by the EU Marine Strategy Framework Directive. Policy responses to plastic waste vary at many levels, ranging from beach clean-up to bans on the commercialization of plastic bags and to Regional Plans for waste management and recycling. Moreover, in recent years, the production of plant-derived biodegradable plastic polymers has assumed increasing importance. This study reports the first preliminary characterization of carbon stable isotopes (δ(13)C) of different plastic polymers (petroleum- and plant-derived) in order to increase the dataset of isotopic values as a tool for further investigation in different fields of polymers research as well as in the marine environment surveillance. The δ(13)C values determined in different packaging for food uses reflect the plant origin of "BIO" materials, whereas the recycled plastic materials displayed a δ(13)C signatures between plant- and petroleum-derived polymers source. In a preliminary estimation, the different colours of plastic did not affect the variability of δ(13)C values, whereas the abiotic and biotic degradation processes that occurred in the plastic materials collected on beaches and in seawater, showed less negative δ(13)C values. A preliminary experimental field test confirmed these results. The advantages offered by isotope ratio mass spectrometry with respect to other analytical methods used to characterize the composition of plastic polymers are: high sensitivity, small amount of material required, rapidity of analysis, low cost and no limitation in black/dark samples compared with spectroscopic analysis.

  12. IMPROVING CONTROL ROOM DESIGN AND OPERATIONS BASED ON HUMAN FACTORS ANALYSES OR HOW MUCH HUMAN FACTORS UPGRADE IS ENOUGH ?

    SciTech Connect

    HIGGINS,J.C.; OHARA,J.M.; ALMEIDA,P.

    2002-09-19

    THE JOSE CABRERA NUCLEAR POWER PLANT IS A ONE LOOP WESTINGHOUSE PRESSURIZED WATER REACTOR. IN THE CONTROL ROOM, THE DISPLAYS AND CONTROLS USED BY OPERATORS FOR THE EMERGENCY OPERATING PROCEDURES ARE DISTRIBUTED ON FRONT AND BACK PANELS. THIS CONFIGURATION CONTRIBUTED TO RISK IN THE PROBABILISTIC SAFETY ASSESSMENT WHERE IMPORTANT OPERATOR ACTIONS ARE REQUIRED. THIS STUDY WAS UNDERTAKEN TO EVALUATE THE IMPACT OF THE DESIGN ON CREW PERFORMANCE AND PLANT SAFETY AND TO DEVELOP DESIGN IMPROVEMENTS.FIVE POTENTIAL EFFECTS WERE IDENTIFIED. THEN NUREG-0711 [1], PROGRAMMATIC, HUMAN FACTORS, ANALYSES WERE CONDUCTED TO SYSTEMATICALLY EVALUATE THE CR-LA YOUT TO DETERMINE IF THERE WAS EVIDENCE OF THE POTENTIAL EFFECTS. THESE ANALYSES INCLUDED OPERATING EXPERIENCE REVIEW, PSA REVIEW, TASK ANALYSES, AND WALKTHROUGH SIMULATIONS. BASED ON THE RESULTS OF THESE ANALYSES, A VARIETY OF CONTROL ROOM MODIFICATIONS WERE IDENTIFIED. FROM THE ALTERNATIVES, A SELECTION WAS MADE THAT PROVIDED A REASONABLEBALANCE BE TWEEN PERFORMANCE, RISK AND ECONOMICS, AND MODIFICATIONS WERE MADE TO THE PLANT.

  13. Incorporating the human gene annotations in different databases significantly improved transcriptomic and genetic analyses.

    PubMed

    Chen, Geng; Wang, Charles; Shi, Leming; Qu, Xiongfei; Chen, Jiwei; Yang, Jianmin; Shi, Caiping; Chen, Long; Zhou, Peiying; Ning, Baitang; Tong, Weida; Shi, Tieliu

    2013-04-01

    Human gene annotation is crucial for conducting transcriptomic and genetic studies; however, the impacts of human gene annotations in diverse databases on related studies have been less evaluated. To enable full use of various human annotation resources and better understand the human transcriptome, here we systematically compare the human annotations present in RefSeq, Ensembl (GENCODE), and AceView on diverse transcriptomic and genetic analyses. We found that the human gene annotations in the three databases are far from complete. Although Ensembl and AceView annotated more genes than RefSeq, more than 15,800 genes from Ensembl (or AceView) are within the intergenic and intronic regions of AceView (or Ensembl) annotation. The human transcriptome annotations in RefSeq, Ensembl, and AceView had distinct effects on short-read mapping, gene and isoform expression profiling, and differential expression calling. Furthermore, our findings indicate that the integrated annotation of these databases can obtain a more complete gene set and significantly enhance those transcriptomic analyses. We also observed that many more known SNPs were located within genes annotated in Ensembl and AceView than in RefSeq. In particular, 1033 of 3041 trait/disease-associated SNPs involved in about 200 human traits/diseases that were previously reported to be in RefSeq intergenic regions could be relocated within Ensembl and AceView genes. Our findings illustrate that a more complete transcriptome generated by incorporating human gene annotations in diverse databases can strikingly improve the overall results of transcriptomic and genetic studies.

  14. Preliminary thermal/thermomechanical analyses of the Site Characterization Plan's Conceptual Design for a repository containing horizontally emplaced waste packages at the Deaf Smith County site

    SciTech Connect

    Ghantous, N.Y.; Raines, G.E.

    1987-10-01

    This report presents thermal/thermomechanical analyses of the Site Characterization Plan Conceptual Design for horizontal package emplacement at the Deaf Smith County site, Texas. The repository was divided into three geometric regions. Then two-dimensional finite-element models were set up to approximate the three-dimensional nature of each region. Thermal and quasistatic thermomechanical finite-element analyses were performed to evaluate the thermal/thermomechanical responses of the three regions. The exponential-time creep law was used to represent the creep behavior of salt rock. The repository design was evaluated by comparing the thermal/thermomechanical responses obtained for the three regions with interim performance constraints. The preliminary results show that all the performance constraints are met except for those of the waste package. The following factors were considered in interpreting these results: (1) the qualitative description of the analytical responses; (2) the limitations of the analyses; and (3) either the conclusions based on overall evaluation of limitations and analytical results or the conclusions based on the fact that the repository design may be evaluated only after further analyses. Furthermore, a parametric analysis was performed to estimate the effect of material parameters on the predicted thermal/thermomechanical response. 23 refs., 34 figs., 9 tabs.

  15. Microgravity and Materials Processing Facility study (MMPF): Requirements and Analyses of Commercial Operations (RACO) preliminary data release

    NASA Technical Reports Server (NTRS)

    1988-01-01

    This requirements and analyses of commercial operations (RACO) study data release reflects the current status of research activities of the Microgravity and Materials Processing Facility under Modification No. 21 to NASA/MSFC Contract NAS8-36122. Section 1 includes 65 commercial space processing projects suitable for deployment aboard the Space Station. Section 2 contains reports of the R:BASE (TM) electronic data base being used in the study, synopses of the experiments, and a summary of data on the experimental facilities. Section 3 is a discussion of video and data compression techniques used as well as a mission timeline analysis.

  16. A preliminary analysis on the dependence of the human diseases on the relative number of sunspot.

    NASA Astrophysics Data System (ADS)

    Ma, Yuehua; Song, Yi

    1996-03-01

    On the basis of the solar-terrestrial relations point of view, the paper investigates the influences of solar activities upon the human race. According to the data of Nanjing Hospital for Infectious Diseases, both the curve of the occurrence of various diseases and the relative number of sunspots with time are drawn, and their related coefficients are calculated. The preliminary results show that the incidences of typhus and scarlet fever keep in step with the 11-year cycle of solar activities, they get the maximum at the same year, while other diseases are not definite.

  17. Phylogenomic analyses reveal convergent patterns of adaptive evolution in elephant and human ancestries.

    PubMed

    Goodman, Morris; Sterner, Kirstin N; Islam, Munirul; Uddin, Monica; Sherwood, Chet C; Hof, Patrick R; Hou, Zhuo-Cheng; Lipovich, Leonard; Jia, Hui; Grossman, Lawrence I; Wildman, Derek E

    2009-12-08

    Specific sets of brain-expressed genes, such as aerobic energy metabolism genes, evolved adaptively in the ancestry of humans and may have evolved adaptively in the ancestry of other large-brained mammals. The recent addition of genomes from two afrotherians (elephant and tenrec) to the expanding set of publically available sequenced mammalian genomes provided an opportunity to test this hypothesis. Elephants resemble humans by having large brains and long life spans; tenrecs, in contrast, have small brains and short life spans. Thus, we investigated whether the phylogenomic patterns of adaptive evolution are more similar between elephant and human than between either elephant and tenrec lineages or human and mouse lineages, and whether aerobic energy metabolism genes are especially well represented in the elephant and human patterns. Our analyses encompassed approximately 6,000 genes in each of these lineages with each gene yielding extensive coding sequence matches in interordinal comparisons. Each gene's nonsynonymous and synonymous nucleotide substitution rates and dN/dS ratios were determined. Then, from gene ontology information on genes with the higher dN/dS ratios, we identified the more prevalent sets of genes that belong to specific functional categories and that evolved adaptively. Elephant and human lineages showed much slower nucleotide substitution rates than tenrec and mouse lineages but more adaptively evolved genes. In correlation with absolute brain size and brain oxygen consumption being largest in elephants and next largest in humans, adaptively evolved aerobic energy metabolism genes were most evident in the elephant lineage and next most evident in the human lineage.

  18. Evidence for Balancing Selection from Nucleotide Sequence Analyses of Human G6PD

    PubMed Central

    Verrelli, Brian C.; McDonald, John H.; Argyropoulos, George; Destro-Bisol, Giovanni; Froment, Alain; Drousiotou, Anthi; Lefranc, Gerard; Helal, Ahmed N.; Loiselet, Jacques; Tishkoff, Sarah A.

    2002-01-01

    Glucose-6-phosphate dehydrogenase (G6PD) mutations that result in reduced enzyme activity have been implicated in malarial resistance and constitute one of the best examples of selection in the human genome. In the present study, we characterize the nucleotide diversity across a 5.2-kb region of G6PD in a sample of 160 Africans and 56 non-Africans, to determine how selection has shaped patterns of DNA variation at this gene. Our global sample of enzymatically normal B alleles and A, A−, and Med alleles with reduced enzyme activities reveals many previously uncharacterized silent-site polymorphisms. In comparison with the absence of amino acid divergence between human and chimpanzee G6PD sequences, we find that the number of G6PD amino acid polymorphisms in human populations is significantly high. Unlike many other G6PD-activity alleles with reduced activity, we find that the age of the A variant, which is common in Africa, may not be consistent with the recent emergence of severe malaria and therefore may have originally had a historically different adaptive function. Overall, our observations strongly support previous genotype-phenotype association studies that proposed that balancing selection maintains G6PD deficiencies within human populations. The present study demonstrates that nucleotide sequence analyses can reveal signatures of both historical and recent selection in the genome and may elucidate the impact that infectious disease has had during human evolution. PMID:12378426

  19. New genetic and linguistic analyses show ancient human influence on baobab evolution and distribution in Australia.

    PubMed

    Rangan, Haripriya; Bell, Karen L; Baum, David A; Fowler, Rachael; McConvell, Patrick; Saunders, Thomas; Spronck, Stef; Kull, Christian A; Murphy, Daniel J

    2015-01-01

    This study investigates the role of human agency in the gene flow and geographical distribution of the Australian baobab, Adansonia gregorii. The genus Adansonia is a charismatic tree endemic to Africa, Madagascar, and northwest Australia that has long been valued by humans for its multiple uses. The distribution of genetic variation in baobabs in Africa has been partially attributed to human-mediated dispersal over millennia, but this relationship has never been investigated for the Australian species. We combined genetic and linguistic data to analyse geographic patterns of gene flow and movement of word-forms for A. gregorii in the Aboriginal languages of northwest Australia. Comprehensive assessment of genetic diversity showed weak geographic structure and high gene flow. Of potential dispersal vectors, humans were identified as most likely to have enabled gene flow across biogeographic barriers in northwest Australia. Genetic-linguistic analysis demonstrated congruence of gene flow patterns and directional movement of Aboriginal loanwords for A. gregorii. These findings, along with previous archaeobotanical evidence from the Late Pleistocene and Holocene, suggest that ancient humans significantly influenced the geographic distribution of Adansonia in northwest Australia.

  20. New Genetic and Linguistic Analyses Show Ancient Human Influence on Baobab Evolution and Distribution in Australia

    PubMed Central

    Rangan, Haripriya; Bell, Karen L.; Baum, David A.; Fowler, Rachael; McConvell, Patrick; Saunders, Thomas; Spronck, Stef; Kull, Christian A.; Murphy, Daniel J.

    2015-01-01

    This study investigates the role of human agency in the gene flow and geographical distribution of the Australian baobab, Adansonia gregorii. The genus Adansonia is a charismatic tree endemic to Africa, Madagascar, and northwest Australia that has long been valued by humans for its multiple uses. The distribution of genetic variation in baobabs in Africa has been partially attributed to human-mediated dispersal over millennia, but this relationship has never been investigated for the Australian species. We combined genetic and linguistic data to analyse geographic patterns of gene flow and movement of word-forms for A. gregorii in the Aboriginal languages of northwest Australia. Comprehensive assessment of genetic diversity showed weak geographic structure and high gene flow. Of potential dispersal vectors, humans were identified as most likely to have enabled gene flow across biogeographic barriers in northwest Australia. Genetic-linguistic analysis demonstrated congruence of gene flow patterns and directional movement of Aboriginal loanwords for A. gregorii. These findings, along with previous archaeobotanical evidence from the Late Pleistocene and Holocene, suggest that ancient humans significantly influenced the geographic distribution of Adansonia in northwest Australia. PMID:25830225

  1. Phylogenetic Analyses Indicate an Atypical Nurse-to-Patient Transmission of Human Immunodeficiency Virus Type 1

    PubMed Central

    Goujon, Christophe P.; Schneider, Véronique M.; Grofti, Jaouad; Montigny, Joëlle; Jeantils, Vincent; Astagneau, Pascal; Rozenbaum, Willy; Lot, Florence; Frocrain-Herchkovitch, Claudie; Delphin, Nathalie; Le Gal, Frédéric; Nicolas, Jean-Claude; Milinkovitch, Michel C.; Dény, Paul

    2000-01-01

    A human immunodeficiency virus (HIV)-negative patient with no risk factor experienced HIV type 1 (HIV-1) primary infection 4 weeks after being hospitalized for surgery. Among the medical staff, only two night shift nurses were identified as HIV-1 seropositive. No exposure to blood was evidenced. To test the hypothesis of a possible nurse-to-patient transmission, phylogenetic analyses were conducted using two HIV-1 genomic regions (pol reverse transcriptase [RT] and env C2C4), each compared with reference strains and large local control sets (57 RT and 41 C2C4 local controls). Extensive analyses using multiple methodologies allowed us to test the robustness of phylogeny inference and to assess transmission hypotheses. Results allow us to unambiguously exclude one HIV-positive nurse and strongly suggest the other HIV-positive nurse as the source of infection of the patient. PMID:10684266

  2. Dimensions of Human-Work Domain Interaction: A Preliminary Analysis for the Design of a Corporate Digital Library.

    ERIC Educational Resources Information Center

    Xie, Hong

    2003-01-01

    Applies the cognitive system engineering approach to investigate human-work interaction at a corporate setting. Reports preliminary analysis of data collected from diary analysis and interview of 20 subjects. Results identify three dimensions for each of four interactive activities involved in human-work interaction and their relationships.…

  3. A preliminary investigation of the fish food web in the Gironde estuary, France, using dietary and stable isotope analyses

    NASA Astrophysics Data System (ADS)

    Pasquaud, S.; Elie, P.; Jeantet, C.; Billy, I.; Martinez, P.; Girardin, M.

    2008-06-01

    Carbon and Nitrogen stable isotopes and stomach contents analyses were used to investigate an estuarine fish food web and identify the contribution of these two methods to the knowledge and understanding of the food web's structure and its functioning. The nine most abundant fish species during the warm period in the Gironde estuary (southwest France, Europe) are examined. Observation of the stomach contents reflects a variety of feeding modes between fish species that consume a diverse assortment of prey, with limited dietary overlap. Nevertheless, when regarding the whole fish community, few prey species dominate the stomach contents. Nitrogen isotope ratios indicate a high intraspecific variability inducing an interspecific covering of the signatures. However, a tendency to δ 15N enrichment according to the trophic position of the species studied was observed. Fish assemblages show a trend towards enrichment of their carbon isotopic signatures from the upper estuary (-20.8 ± 1.8‰) towards the lower estuary (-18.3 ± 1.6‰). But whatever the capture zone considered, most of the individual δ 13C values for each fish analysed are comprised between -22 and -16‰. Only few specimens, belonging to migratory amphihaline species, have significantly lighter values. The stomach contents method of analysis has the advantage of giving an initial view of the ichthyological trophic structure of the system by describing the food relations between a fish species and its prey. From these results, hypotheses can be drawn about the network's functioning, suggesting a sharing of resources between species and a "wasp-waist" control of this estuarine food web. The stable isotope analysis method enables us to improve our structural knowledge by positioning the different species in a food web, with their position being determined by the number of energy transfers (analysis of δ 15N). Conversely, in environments as complex and changing as estuaries, it appears difficult to

  4. Detection and preliminary screening of the human gene expression profile for Hirschsprung's disease

    PubMed Central

    WANG, XIN; WANG, SHIQI; JIN, XIANQING; WANG, NING; LUO, YUANYUAN; TENG, YINPING

    2016-01-01

    The present study investigated a genome microarray of colorectal lesions (spasm segments) in children with Hirschsprung's disease (HSCR), and analyzed the results. In addition, the present study screened for differentially expressed genes in children with HSCR. Microarray technology was used to examine the human gene expression profiles of the colorectal lesions (spasm segments) of six children with HSCR, and three normal colon tissue samples. The data were analyzed be determining P-values of significance and absolute fold changes. Preliminary screening was performed to identify genes exhibiting significant differential expression in children with HSCR, and these target genes were analyzed in subsequent verification and analytical investigations. Of >20,000 detected human genes, the preliminary screenings demonstrated that 3,850 genes were differentially expressed and upregulated, with P<0.05 and >2-fold absolute changes in expression. In addition, 645 differentially expressed genes with P<0.05 and >2-fold absolute changes were downregulated. Of the upregulated genes, 118 were involved in classic signaling pathways, compared with 11 of the downregulated genes (P<0.001; absolute fold change >2-fold). HSCR etiology is complex and often involves multiple gene changes. Microarray technology can produce large quantities of gene expression data simultaneously, and analyzing this data using various techniques may provide a fast and efficient method for identifying novel gene targets and for investigating the mechanisms underlying HSCR pathogenesis. PMID:26648025

  5. Preliminary results of an assessment of FGGE 'special effort' data and its impact on GLAS model analyses and forecasts

    NASA Technical Reports Server (NTRS)

    Atlas, R.

    1981-01-01

    Attempts to improve the handling of satellite temperature and cloud-derived data during the FGGE experiment are compared with results gained during the trials. The satellite data were edited by assigning quality indicators to temperature soundings by means of the IR TV camera on the Tiros-N spacecraft, and to cloud-tracked wind data from GEO satellites. The data quality were assayed on the basis of horizontal, vertical, and temporal consistency of observed synoptic conditions. The data interpreter-operator retrieved all available data for an area, determined which regions were data-sparse, then used data-rich areas nearby to supply smoothed gradients into the zones with insufficient data. Results are presented for measurements taken during Jan.-Feb., and May 1979 over Alaska. It was found that enhanced and operational IR soundings were mostly equivalent. Some improvements were implemented in the 1000-300 mb layer thickness and intensification of atmospheric thermal gradients. Interactive processing was concluded to be an aid to processing Tiros-N data. The data-improvements treatment was found to significantly modify large scale analyses.

  6. Naturally induced secondary radiation in interplanetary space: Preliminary analyses for gamma radiation and radioisotope production from thermal neutron activation

    NASA Astrophysics Data System (ADS)

    Plaza-Rosado, Heriberto

    1991-09-01

    Thermal neutron activation analyses were carried out for various space systems components to determine gamma radiation dose rates and food radiation contamination levels. The space systems components selected were those for which previous radiation studies existed. These include manned space vehicle radiation shielding, liquid hydrogen propellant tanks for a Mars mission, and a food supply used as space vehicle radiation shielding. The computational method used is based on the fast neutron distribution generated by the BRYNTRN and HZETRN transport codes for Galactic Cosmic Rays (GCR) at solar minimum conditions and intense solar flares in space systems components. The gamma dose rates for soft tissue are calculated for water and aluminum space vehicle slab shields considering volumetric source self-attenuation and exponential buildup factors. In the case of the lunar habitat with regolith shielding, a completely exposed spherical habitat was assumed for mathematical convenience and conservative calculations. Activation analysis of the food supply used as radiation shielding is presented for four selected nutrients: potassium, calcium, sodium, and phosphorus. Radioactive isotopes that could represent a health hazard if ingested are identified and their concentrations are identified. For nutrients soluble in water, it was found that all induced radioactivity was below the accepted maximum permissible concentrations.

  7. Naturally induced secondary radiation in interplanetary space: Preliminary analyses for gamma radiation and radioisotope production from thermal neutron activation

    NASA Technical Reports Server (NTRS)

    Plaza-Rosado, Heriberto

    1991-01-01

    Thermal neutron activation analyses were carried out for various space systems components to determine gamma radiation dose rates and food radiation contamination levels. The space systems components selected were those for which previous radiation studies existed. These include manned space vehicle radiation shielding, liquid hydrogen propellant tanks for a Mars mission, and a food supply used as space vehicle radiation shielding. The computational method used is based on the fast neutron distribution generated by the BRYNTRN and HZETRN transport codes for Galactic Cosmic Rays (GCR) at solar minimum conditions and intense solar flares in space systems components. The gamma dose rates for soft tissue are calculated for water and aluminum space vehicle slab shields considering volumetric source self-attenuation and exponential buildup factors. In the case of the lunar habitat with regolith shielding, a completely exposed spherical habitat was assumed for mathematical convenience and conservative calculations. Activation analysis of the food supply used as radiation shielding is presented for four selected nutrients: potassium, calcium, sodium, and phosphorus. Radioactive isotopes that could represent a health hazard if ingested are identified and their concentrations are identified. For nutrients soluble in water, it was found that all induced radioactivity was below the accepted maximum permissible concentrations.

  8. Preliminary performance assessment for the Waste Isolation Pilot Plant, December 1992. Volume 5, Uncertainty and sensitivity analyses of gas and brine migration for undisturbed performance

    SciTech Connect

    Not Available

    1993-08-01

    Before disposing of transuranic radioactive waste in the Waste Isolation Pilot Plant (WIPP), the United States Department of Energy (DOE) must evaluate compliance with applicable long-term regulations of the United States Environmental Protection Agency (EPA). Sandia National Laboratories is conducting iterative performance assessments (PAs) of the WIPP for the DOE to provide interim guidance while preparing for a final compliance evaluation. This volume of the 1992 PA contains results of uncertainty and sensitivity analyses with respect to migration of gas and brine from the undisturbed repository. Additional information about the 1992 PA is provided in other volumes. Volume 1 contains an overview of WIPP PA and results of a preliminary comparison with 40 CFR 191, Subpart B. Volume 2 describes the technical basis for the performance assessment, including descriptions of the linked computational models used in the Monte Carlo analyses. Volume 3 contains the reference data base and values for input parameters used in consequence and probability modeling. Volume 4 contains uncertainty and sensitivity analyses with respect to the EPA`s Environmental Standards for the Management and Disposal of Spent Nuclear Fuel, High-Level and Transuranic Radioactive Wastes (40 CFR 191, Subpart B). Finally, guidance derived from the entire 1992 PA is presented in Volume 6. Results of the 1992 uncertainty and sensitivity analyses indicate that, conditional on the modeling assumptions and the assigned parameter-value distributions, the most important parameters for which uncertainty has the potential to affect gas and brine migration from the undisturbed repository are: initial liquid saturation in the waste, anhydrite permeability, biodegradation-reaction stoichiometry, gas-generation rates for both corrosion and biodegradation under inundated conditions, and the permeability of the long-term shaft seal.

  9. A preliminary report on noble gas isotope analyses using the Helix-MC multi-collector mass spectrometer

    NASA Astrophysics Data System (ADS)

    Honda, M.; Zhang, X.; Phillips, D.; Szczepanski, S.; Deerberg, M.; Hamilton, D.; Krummen, M.; Schwieters, J.

    2013-12-01

    Analyses of noble gas isotopes by multi-collector mass spectrometry substantially improve measurement precision and accuracy, with the potential to revolutionise applications to cosmo-geo-sciences. The Helix-MC noble gas mass spectrometer manufactured by Thermo-Fisher is a 350mm, 120 degree extended geometry, high resolution, multi-collector mass spectrometer for the simultaneous analysis of noble gas isotopes. The detector array includes a fixed axial (Ax) detector, 2 adjustable high mass (H1 and H2) detectors and 2 adjustable low mass (L1 and L2) detectors. Each detector is equipped with a Faraday/ion counting multiplier CFM (Combined Faraday and CDD Multiplier) detector. Mass resolution and mass resolving power on the H2, Ax and L2 detectors of the Helix-MC installed at the Australian National University (ANU) are approximately 1,800 and 8,000, respectively. The noble gas handling system on-line to the Helix-MC consists of: (1) a resistively-heated, double-vacuum, tantalum furnace system, (2) air actuated vacuum crusher, (3) Photon-Machines diode laser heating system, (4) Janis He cryogenic trap assembly, (5) gas purification system and (6) standard gas pipette tanks, which are totally automated and controlled by the Qtegra software platform developed by Thermo-Fisher. Eleven repeat measurements of atmospheric Ar using the H2 Faraday (1E11 ohm resistor) and L2 CDD collectors on the Helix-MC, yield a mean 40Ar/36Ar ratio of 322.09 +- 0.28 (0.089%) with a 4,700 fA 40Ar beam current. This result compares favourably with the precision achieved by the Argus VI at the University of Melbourne (318.12 +- 0.17; 0.052%; n = 10) with a similar beam size of 4,200 fA. The high mass resolution of the L2 collector permits complete separation of the 36Ar and interfering 3 x 12C (required mass resolution (MR) of 1,100) and partial separation of H35Cl (MR = 3,900). This capability enables evaluation of the significance of Ar isotopic interferences related to the correction of

  10. Joint analyses model for total cholesterol and triglyceride in human serum with near-infrared spectroscopy

    NASA Astrophysics Data System (ADS)

    Yao, Lijun; Lyu, Ning; Chen, Jiemei; Pan, Tao; Yu, Jing

    2016-04-01

    The development of a small, dedicated near-infrared (NIR) spectrometer has promising potential applications, such as for joint analyses of total cholesterol (TC) and triglyceride (TG) in human serum for preventing and treating hyperlipidemia of a large population. The appropriate wavelength selection is a key technology for developing such a spectrometer. For this reason, a novel wavelength selection method, named the equidistant combination partial least squares (EC-PLS), was applied to the wavelength selection for the NIR analyses of TC and TG in human serum. A rigorous process based on the various divisions of calibration and prediction sets was performed to achieve modeling optimization with stability. By applying EC-PLS, a model set was developed, which consists of various models that were equivalent to the optimal model. The joint analyses model of the two indicators was further selected with only 50 wavelengths. The random validation samples excluded from the modeling process were used to validate the selected model. The root-mean-square errors, correlation coefficients and ratio of performance to deviation for the prediction were 0.197 mmol L- 1, 0.985 and 5.6 for TC, and 0.101 mmol L- 1, 0.992 and 8.0 for TG, respectively. The sensitivity and specificity for hyperlipidemia were 96.2% and 98.0%. These findings indicate high prediction accuracy and low model complexity. The proposed wavelength selection provided valuable references for the designing of a small, dedicated spectrometer for hyperlipidemia. The methodological framework and optimization algorithm are universal, such that they can be applied to other fields.

  11. Preliminary perspectives on DNA collection in anti-human trafficking efforts.

    PubMed

    Katsanis, Sara H; Kim, Joyce; Minear, Mollie A; Chandrasekharan, Subhashini; Wagner, Jennifer K

    2014-01-01

    Forensic DNA methodologies have potential applications in the investigation of human trafficking cases. DNA and relationship testing may be useful for confirmation of biological relationship claims in immigration, identification of trafficked individuals who are missing persons, and family reunification of displaced individuals after mass disasters and conflicts. As these applications rely on the collection of DNA from non-criminals and potentially vulnerable individuals, questions arise as to how to address the ethical challenges of collection, security, and privacy of collected samples and DNA profiles. We administered a survey targeted to victims' advocates to gain preliminary understanding of perspectives regarding human trafficking definitions, DNA and sex workers, and perceived trust of authorities potentially involved in DNA collection. We asked respondents to consider the use of DNA for investigating adoption fraud, sex trafficking, and post-conflict child soldier cases. We found some key differences in perspectives on defining what qualifies as "trafficking." When we varied terminology between "sex worker" and "sex trafficking victim" we detected differences in perception on which authorities can be trusted. Respondents were supportive of the hypothetical models proposed to collect DNA. Most were favorable of DNA specimens being controlled by an authority outside of law enforcement. Participants voiced concerns focused on privacy, misuse of DNA samples and data, unintentional harms, data security, and infrastructure. These preliminary data indicate that while there is perceived value in programs to use DNA for investigating cases of human trafficking, these programs may need to consider levels of trust in authorities as their logistics are developed and implemented.

  12. Molecular analyses of neurogenic defects in a human pluripotent stem cell model of fragile X syndrome.

    PubMed

    Boland, Michael J; Nazor, Kristopher L; Tran, Ha T; Szücs, Attila; Lynch, Candace L; Paredes, Ryder; Tassone, Flora; Sanna, Pietro Paolo; Hagerman, Randi J; Loring, Jeanne F

    2017-03-01

    New research suggests that common pathways are altered in many neurodevelopmental disorders including autism spectrum disorder; however, little is known about early molecular events that contribute to the pathology of these diseases. The study of monogenic, neurodevelopmental disorders with a high incidence of autistic behaviours, such as fragile X syndrome, has the potential to identify genes and pathways that are dysregulated in autism spectrum disorder as well as fragile X syndrome. In vitro generation of human disease-relevant cell types provides the ability to investigate aspects of disease that are impossible to study in patients or animal models. Differentiation of human pluripotent stem cells recapitulates development of the neocortex, an area affected in both fragile X syndrome and autism spectrum disorder. We have generated induced human pluripotent stem cells from several individuals clinically diagnosed with fragile X syndrome and autism spectrum disorder. When differentiated to dorsal forebrain cell fates, our fragile X syndrome human pluripotent stem cell lines exhibited reproducible aberrant neurogenic phenotypes. Using global gene expression and DNA methylation profiling, we have analysed the early stages of neurogenesis in fragile X syndrome human pluripotent stem cells. We discovered aberrant DNA methylation patterns at specific genomic regions in fragile X syndrome cells, and identified dysregulated gene- and network-level correlates of fragile X syndrome that are associated with developmental signalling, cell migration, and neuronal maturation. Integration of our gene expression and epigenetic analysis identified altered epigenetic-mediated transcriptional regulation of a distinct set of genes in fragile X syndrome. These fragile X syndrome-aberrant networks are significantly enriched for genes associated with autism spectrum disorder, giving support to the idea that underlying similarities exist among these neurodevelopmental diseases.

  13. Full genomic analyses of human rotavirus strains possessing the rare P[8]b VP4 subtype.

    PubMed

    Ghosh, Souvik; Paul, Shyamal Kumar; Yamamoto, Dai; Nagashima, Shigeo; Kobayashi, Nobumichi

    2011-08-01

    Rotaviruses with the P[8] VP4 genotype are a major cause of acute infantile diarrhea. The P[8] genotype is classified into two genetically distinct subtypes, P[8]a and P[8]b. Most of the P[8] strains belong to subtype P[8]a, whilst P[8]b strains are rare. To date, the whole genomes of a few P[8]a strains have been analyzed, whilst there are no reports on full genomic analysis of the P[8]b strains. To determine the genetic makeup of the rare P[8]b strains and their overall genetic relatedness to the P[8]a strains, the present study analyzed the full genomes of a human G9P[8]b strain, MMC38, and a G1P[8]b strain, MMC71, detected in Bangladesh in 2005. By nucleotide sequence identities and phylogenetic analyses, strains MMC38 and MMC71 exhibited a human rotavirus Wa-like genotype constellation. Except for the VP4 gene, all the genes of strains MMC38 and MMC71 were closely related to cognate genes of the contemporary and more recent human Wa-like G1P[8]a, G9P[8]a, G11P[8]a, G11P[25], G12P[6] and G12P[8]a strains, including those from Bangladesh. Therefore, strains MMC38 and MMC71 possessed the genetically distinct P[8]b VP4 gene on a common human Wa-like genetic backbone, pointing towards their possible origin from reassortment events between common human Wa-like strains and unidentified rotavirus strains possessing the rare P[8]b-like VP4 gene. Since strains with this stable Wa-like genetic backbone can spread rapidly, and it is not certain as to whether the current rotavirus vaccines will be equally efficacious against the P[8]b strains as the P[8]a strains, proper detection of P[8]b strains and their whole genomic analyses might be of public health significance. To our knowledge, the present study is the first report on full genomic analysis of the rare P[8]b rotavirus strains.

  14. Multiple Skin Cancers in a Renal Transplant Recipient: A Patient Report with Analyses of Human Papillomavirus and Human Polyomavirus Infection

    PubMed Central

    Kaneda, Tokinobu; Matsushita, Michiko; Iwasaki, Takeshi; Ishiguro, Naoko; Koide, Takashi; Hayashi, Kazuhiko; Kitamura, Yukisato

    2015-01-01

    Skin cancer is an important complication in renal transplant recipients. Associations of transplant-related skin tumor with ultraviolet radiation, age at transplantation, type of immunosuppressant drug administered, and viral infection have been reported; however, the details remain unclear. We report a 61-year-old man who had underwent renal transplantation at 38 years of age and developed multiple skin tumors or squamous cell carcinomas (SCCs). Polymerase chain reaction (PCR) analyses of the patient’s 12 tumors for viral DNAs of cutaneous or mucosal human papillomavirus (HPV) and 6 human polyomaviruses (MCPyV, trichodysplasia spinulosa-associated, BK, JC, KI and WU polyomaviruses) only detected cutaneous HPV-DNA in only 5 of the tumors; no other viruses were detected. Real-time PCR showed high loads of cutaneous HPV in 3 SCCs and very low loads of MCPyV in 9. Immunohistochemistry revealed no tumor cell expression for MCPyV-large T-antigen or mucosal HPV. Our report not only reconfirmed the association of cutaneous HPV5 with skin cancer in renal transplant recipients in previous studies but also showed no relevant association of 6 human polyomaviruses and mucosal HPV with skin tumors. PMID:26538801

  15. Gene expression analyses of the spatio-temporal relationships of human medulloblastoma subgroups during early human neurogenesis.

    PubMed

    Hooper, Cornelia M; Hawes, Susan M; Kees, Ursula R; Gottardo, Nicholas G; Dallas, Peter B

    2014-01-01

    Medulloblastoma is the most common form of malignant paediatric brain tumour and is the leading cause of childhood cancer related mortality. The four molecular subgroups of medulloblastoma that have been identified - WNT, SHH, Group 3 and Group 4 - have molecular and topographical characteristics suggestive of different cells of origin. Definitive identification of the cell(s) of origin of the medulloblastoma subgroups, particularly the poorer prognosis Group 3 and Group 4 medulloblastoma, is critical to understand the pathogenesis of the disease, and ultimately for the development of more effective treatment options. To address this issue, the gene expression profiles of normal human neural tissues and cell types representing a broad neuro-developmental continuum, were compared to those of two independent cohorts of primary human medulloblastoma specimens. Clustering, co-expression network, and gene expression analyses revealed that WNT and SHH medulloblastoma may be derived from distinct neural stem cell populations during early embryonic development, while the transcriptional profiles of Group 3 and Group 4 medulloblastoma resemble cerebellar granule neuron precursors at weeks 10-15 and 20-30 of embryogenesis, respectively. Our data indicate that Group 3 medulloblastoma may arise through abnormal neuronal differentiation, whereas deregulation of synaptic pruning-associated apoptosis may be driving Group 4 tumorigenesis. Overall, these data provide significant new insight into the spatio-temporal relationships and molecular pathogenesis of the human medulloblastoma subgroups, and provide an important framework for the development of more refined model systems, and ultimately improved therapeutic strategies.

  16. Gene Expression Analyses of the Spatio-Temporal Relationships of Human Medulloblastoma Subgroups during Early Human Neurogenesis

    PubMed Central

    Hooper, Cornelia M.; Hawes, Susan M.; Kees, Ursula R.; Gottardo, Nicholas G.; Dallas, Peter B.

    2014-01-01

    Medulloblastoma is the most common form of malignant paediatric brain tumour and is the leading cause of childhood cancer related mortality. The four molecular subgroups of medulloblastoma that have been identified – WNT, SHH, Group 3 and Group 4 - have molecular and topographical characteristics suggestive of different cells of origin. Definitive identification of the cell(s) of origin of the medulloblastoma subgroups, particularly the poorer prognosis Group 3 and Group 4 medulloblastoma, is critical to understand the pathogenesis of the disease, and ultimately for the development of more effective treatment options. To address this issue, the gene expression profiles of normal human neural tissues and cell types representing a broad neuro-developmental continuum, were compared to those of two independent cohorts of primary human medulloblastoma specimens. Clustering, co-expression network, and gene expression analyses revealed that WNT and SHH medulloblastoma may be derived from distinct neural stem cell populations during early embryonic development, while the transcriptional profiles of Group 3 and Group 4 medulloblastoma resemble cerebellar granule neuron precursors at weeks 10–15 and 20–30 of embryogenesis, respectively. Our data indicate that Group 3 medulloblastoma may arise through abnormal neuronal differentiation, whereas deregulation of synaptic pruning-associated apoptosis may be driving Group 4 tumorigenesis. Overall, these data provide significant new insight into the spatio-temporal relationships and molecular pathogenesis of the human medulloblastoma subgroups, and provide an important framework for the development of more refined model systems, and ultimately improved therapeutic strategies. PMID:25412507

  17. Transcriptomic analyses of the anti-adipogenic effects of oleuropein in human mesenchymal stem cells.

    PubMed

    Casado-Díaz, Antonio; Anter, Jaouad; Müller, Sören; Winter, Peter; Quesada-Gómez, José Manuel; Dorado, Gabriel

    2017-03-22

    Extra virgin olive oil has positive effects on health. Oleuropein is a polyphenolic compound present in olive-tree leaves, fruits (olives) and olive oil. It is responsible for the relevant organoleptic and biological properties of olive oil, including antiadipogenic properties. Thus, the effects of oleuropein on the adipogenesis of human bone-marrow mesenchymal stem cells were studied by transcriptomics and differential gene-expression analyses. Oleuropein could upregulate expression of 60% of adipogenesis-repressed genes. Besides, it could activate signaling pathways such as Rho and β-catenin, maintaining cells at an undifferentiated stage. Our data suggest that mitochondrial activity is reduced by oleuropein, mostly during adipogenic differentiation. These results shed light on oleuropein activity on cells, with potential application as a "nutraceutical" for the prevention and treatment of diseases such as obesity and osteoporosis.

  18. Expression, purification, crystallization, and preliminary X-Ray analysis of the human UDP-glucose dehydrogenase.

    PubMed

    Huh, Jae Wan; Robinson, Robert Charles; Lee, Han Sam; Lee, Jae Il; Heo, Yong Seok; Kim, Hyun Tae; Lee, Hyun Ju; Cho, Sung Woo; Choe, Han

    2006-01-01

    UDP-glucose dehydrogenase (UGDH) catalyzes the synthesis of UDP-glucuronic acid from UDP-glucose resulting in the formation of proteoglycans that are involved in promoting normal cellular growth and migration. Overproduction of proteoglycans has been implicated in the progression of certain epithelial cancers. Here, human UGDH (hUGDH) was purified and crystallized from a solution of 0.2 M ammonium sulfate, 0.1 M Na cacodylate, pH 6.5, and 21% PEG 8000. Diffraction data were collected to a resolution of 2.8 A. The crystal belongs to the orthorhombic space group P2(1)2(1)2(1) with unit-cell parameters a = 173.25, b = 191.16, c = 225.94 A, and alpha = beta = gamma = 90.0 degrees. Based on preliminary analysis of the diffraction data, we propose that the biological unit of hUGDH is a tetramer.

  19. Emergence and evolutionary analysis of the human DDR network: implications in comparative genomics and downstream analyses.

    PubMed

    Arcas, Aida; Fernández-Capetillo, Oscar; Cases, Ildefonso; Rojas, Ana M

    2014-04-01

    The DNA damage response (DDR) is a crucial signaling network that preserves the integrity of the genome. This network is an ensemble of distinct but often overlapping subnetworks, where different components fulfill distinct functions in precise spatial and temporal scenarios. To understand how these elements have been assembled together in humans, we performed comparative genomic analyses in 47 selected species to trace back their emergence using systematic phylogenetic analyses and estimated gene ages. The emergence of the contribution of posttranslational modifications to the complex regulation of DDR was also investigated. This is the first time a systematic analysis has focused on the evolution of DDR subnetworks as a whole. Our results indicate that a DDR core, mostly constructed around metabolic activities, appeared soon after the emergence of eukaryotes, and that additional regulatory capacities appeared later through complex evolutionary process. Potential key posttranslational modifications were also in place then, with interacting pairs preferentially appearing at the same evolutionary time, although modifications often led to the subsequent acquisition of new targets afterwards. We also found extensive gene loss in essential modules of the regulatory network in fungi, plants, and arthropods, important for their validation as model organisms for DDR studies.

  20. Investigation of human exposure to triclocarban after showering and preliminary evaluation of its biological effects.

    PubMed

    Schebb, Nils Helge; Inceoglu, Bora; Ahn, Ki Chang; Morisseau, Christophe; Gee, Shirley J; Hammock, Bruce D

    2011-04-01

    The antibacterial soap additive triclocarban (TCC) is widely used in personal care products. TCC has a high environmental persistence. We developed and validated a sensitive online solid-phase extraction-LC-MS/MS method to rapidly analyze TCC and its major metabolites in urine and other biological samples to assess human exposure. We measured human urine concentrations 0-72 h after showering with a commercial bar soap containing 0.6% TCC. The major route of renal elimination was excretion as N-glucuronides. The absorption was estimated at 0.6% of the 70±15 mg of TCC in the soap used. The TCC-N-glucuronide urine concentration varied widely among the subjects, and continuous daily use of the soap led to steady state levels of excretion. In order to assess potential biological effects arising from this exposure, we screened TCC for the inhibition of human enzymes in vitro. We demonstrate that TCC is a potent inhibitor of the enzyme soluble epoxide hydrolase (sEH), whereas TCC's major metabolites lack strong inhibitory activity. Topical administration of TCC at similar levels to rats in a preliminary in vivo study, however, failed to alter plasma biomarkers of sEH activity. Overall the analytical strategy described here revealed that use of TCC soap causes exposure levels that warrant further evaluation.

  1. Building-related symptoms among U.S. office workers and risks factors for moisture and contamination: Preliminary analyses of U.S. EPA BASE Data

    SciTech Connect

    Mendell, Mark J.; Cozen, Myrna

    2002-09-01

    dirty cooling coils were associated with a nonsignificant increase in lower respiratory symptoms. These preliminary findings suggest that some factors that indicate risks for moisture or contamination in office buildings may have adverse effects on respiratory or neurologic health of office workers. More refined analyses are underway that will include these risk factors in simultaneous multivariate models along with additional risk factors that may be confounders, such as ventilation rate and indoor temperature. Future analyses will also use more refined metrics for both health outcomes and environmental risks, as well as assess risk in susceptible sub-groups.

  2. Computational analyses of mammalian lactate dehydrogenases: human, mouse, opossum and platypus LDHs.

    PubMed

    Holmes, Roger S; Goldberg, Erwin

    2009-10-01

    Computational methods were used to predict the amino acid sequences and gene locations for mammalian lactate dehydrogenase (LDH) genes and proteins using genome sequence databanks. Human LDHA, LDHC and LDH6A genes were located in tandem on chromosome 11, while LDH6B and LDH6C genes were on chromosomes 15 and 12, respectively. Opossum LDHC and LDH6B genes were located in tandem with the opossum LDHA gene on chromosome 5 and contained 7 (LDHA and LDHC) or 8 (LDH6B) exons. An amino acid sequence prediction for the opossum LDH6B subunit gave an extended N-terminal sequence, similar to the human and mouse LDH6B sequences, which may support the export of this enzyme into mitochondria. The platypus genome contained at least 3 LDH genes encoding LDHA, LDHB and LDH6B subunits. Phylogenetic studies and sequence analyses indicated that LDHA, LDHB and LDH6B genes are present in all mammalian genomes examined, including a monotreme species (platypus), whereas the LDHC gene may have arisen more recently in marsupial mammals.

  3. Molecular cytogenetic analysis of human blastocysts andcytotrophoblasts by multi-color FISH and Spectra Imaging analyses

    SciTech Connect

    Weier, Jingly F.; Ferlatte, Christy; Baumgartner, Adolf; Jung,Christine J.; Nguyen, Ha-Nam; Chu, Lisa W.; Pedersen, Roger A.; Fisher,Susan J.; Weier, Heinz-Ulrich G.

    2006-02-08

    Numerical chromosome aberrations in gametes typically lead to failed fertilization, spontaneous abortion or a chromosomally abnormal fetus. By means of preimplantation genetic diagnosis (PGD), we now can screen human embryos in vitro for aneuploidy before transferring the embryos to the uterus. PGD allows us to select unaffected embryos for transfer and increases the implantation rate in in vitro fertilization programs. Molecular cytogenetic analyses using multi-color fluorescence in situ hybridization (FISH) of blastomeres have become the major tool for preimplantation genetic screening of aneuploidy. However, current FISH technology can test for only a small number of chromosome abnormalities and hitherto failed to increase the pregnancy rates as expected. We are in the process of developing technologies to score all 24 chromosomes in single cells within a 3 day time limit, which we believe is vital to the clinical setting. Also, human placental cytotrophoblasts (CTBs) at the fetal-maternal interface acquire aneuploidies as they differentiate to an invasive phenotype. About 20-50% of invasive CTB cells from uncomplicated pregnancies were found aneuploidy, suggesting that the acquisition of aneuploidy is an important component of normal placentation, perhaps limiting the proliferative and invasive potential of CTBs. Since most invasive CTBs are interphase cells and possess extreme heterogeneity, we applied multi-color FISH and repeated hybridizations to investigate individual CTBs. In summary, this study demonstrates the strength of Spectral Imaging analysis and repeated hybridizations, which provides a basis for full karyotype analysis of single interphase cells.

  4. Do Lambs Perceive Regular Human Stroking as Pleasant? Behavior and Heart Rate Variability Analyses

    PubMed Central

    Coulon, Marjorie; Nowak, Raymond; Peyrat, Julie; Chandèze, Hervé; Boissy, Alain; Boivin, Xavier

    2015-01-01

    Stroking by humans is beneficial to the human-animal relationship and improves welfare in many species that express intraspecific allogrooming, but very few studies have looked at species like sheep that do not express such contact except around parturition. This study investigated the way lambs perceive regular human tactile contact using behavioral and physiological responses. Twenty-four lambs were reared and bucket-fed in groups of four. All were stroked daily by their familiar caregiver. At 8 weeks of age, the lambs were individually tested in their home pen but in a 1×1m open-barred pen after a 15h period of habituation to physical separation from peers while remaining in visual and auditory contact. Half of the lambs received stroking by their caregiver for 8min and half were exposed to their caregiver’s immobile presence. Heart rate and heart rate variability were recorded and analyzed by 2-min slots over the same interval based on three measures: mean heart rate value (HR), root mean square of successive differences (RMSSD) and standard deviation of all intervals measured between consecutive sinus beats (SDNN). Behavioral responses (ear postures of the lamb and time spent in contact with the familiar caregiver, on the knees of the familiar caregiver, and moving) were recorded throughout the test. Lamb HR decreased continuously while in the presence of their caregiver. Lambs being stroked showed slower HR and higher RMSSD which reflected positive emotional states compared to lambs left unstroked. All behavioral variables were highly correlated with the main component axis of the PCA analyses: the more the animals stayed in contact with their caregiver, the less they moved and the more their ears were hanging. This first component clearly differentiates lambs being stroked or not. Behavioral and physiological observations support the hypothesis that gentle physical contact with the caregiver is perceived positively by lambs. PMID:25714604

  5. Next Generation Munitions Handler: Human-Machine Interface and Preliminary Performance Evaluation

    SciTech Connect

    Draper, J.V.; Jansen, J.F.; Pin, F.G.; Rowe, J.C.

    1999-04-25

    The Next Generation Munitions Handler/Advanced Technology Demonstrator (NGMI-VATTD) is a technology demonstrator for the application of an advanced robotic device for re-arming U.S. Air Force (USAF) and U.S. Navy (USN) tactical fighters. It comprises two key hardware components: a heavy-lift dexterous manipulator (HDM) and a nonholonomic mobility platform. The NGMWATTD is capable of lifting weapons up to 4400 kg (2000 lb) and placing them on any weapons rack on existing fighters (including the F-22 Raptor). This report describes the NGMH mission with particular reference to human-machine interfaces. It also describes preliminary testing to garner feedback about the heavy-lift manipulator arm from experienced fighter load crewmen. The purpose of the testing was to provide preliminary information about control system parameters and to gather feed- back from users about manipulator arm functionality. To that end, the Air Force load crewmen interacted with the NGMWATTD in an informal testing session and provided feedback about the performance of the system. Certain con- trol system parameters were changed during the course of the testing and feedback from the participants was used to make a rough estimate of "good" initial operating parameters. Later, formal testing will concentrate within this range to identify optimal operating parameters. User reactions to the HDM were generally positive, All of the USAF personnel were favorably impressed with the capabilities of the system. Fine-tuning operating parameters created a system even more favorably regarded by the load crews. Further adjustment to control system parameters will result in a system that is operationally efficient, easy to use, and well accepted by users.

  6. Purification, crystallization and preliminary crystallographic analysis of human cystathionine β-synthase

    PubMed Central

    Oyenarte, Iker; Majtan, Tomas; Ereño, June; Corral-Rodríguez, María Angeles; Kraus, Jan P.; Martínez-Cruz, Luis Alfonso

    2012-01-01

    Human cystathionine β-synthase (CBS) is a pyridoxal-5′-phosphate-dependent hemeprotein, whose catalytic activity is regulated by S-adenosylmethionine. CBS catalyzes the β-replacement reaction of homocysteine (Hcy) with serine to yield cystathionine. CBS is a key regulator of plasma levels of the thrombogenic Hcy and deficiency in CBS is the single most common cause of homocystinuria, an inherited metabolic disorder of sulfur amino acids. The properties of CBS enzymes, such as domain organization, oligomerization degree or regulatory mechanisms, are not conserved across the eukaryotes. The current body of knowledge is insufficient to understand these differences and their impact on CBS function and physiology. To overcome this deficiency, we have addressed the crystallization and preliminary crystallographic analysis of a protein construct (hCBS516–525) that contains the full-length CBS from Homo sapiens (hCBS) and just lacks amino-acid residues 516–525, which are located in a disordered loop. The human enzyme yielded crystals belonging to space group I222, with unit-cell parameters a = 124.98, b = 136.33, c = 169.83 Å and diffracting X-rays to a resolution of 3.0 Å. The crystal structure appears to contain two molecules in the asymmetric unit which presumably correspond to a dimeric form of the enzyme. PMID:23143240

  7. Preliminary study of cytotoxic effects of photodynamic therapy and immunotherapy on human pancreatic cancer cells

    NASA Astrophysics Data System (ADS)

    Wang, Luowei; Liu, Bolin; Chen, Yang K.; Li, Zhaoshen; Hetzel, Fred W.; Huang, Zheng

    2009-02-01

    Pancreatic cancer is the fourth most common cause of cancer death in the western world. The disease is very resistant to radiotherapy and chemotherapy. One reason for that is the resistance of pancreatic cancer cells to apoptosis. Among the current investigational approaches, targeting human epidermal growth factor receptor (HER-1/EGFR) and interstitial photodynamic therapy (PDT) show promises. When used alone or together, these new approaches might provide an alternative modality to treat pancreatic cancer. This study examined and compared cytotoxic effects of antibody C225 (an anti-HER-1/EGFR monoclonal antibody) and Photofrin-mediated PDT on two human pancreatic cancer cell lines (BxPc-3, HPAF-II). Preliminary in vitro data indicated that these treatments could block various proliferation pathways of pancreatic cancer cells through different mechanisms. For instance, PDT could induce early apoptosis. C225 could induce G1 arrest. These findings might help to design new strategies such as the combination of PDT and immunotherapy for the treatment of pancreatic cancer.

  8. Analyses of the stability and core taxonomic memberships of the human microbiome.

    PubMed

    Li, Kelvin; Bihan, Monika; Methé, Barbara A

    2013-01-01

    Analyses of the taxonomic diversity associated with the human microbiome continue to be an area of great importance. The study of the nature and extent of the commonly shared taxa ("core"), versus those less prevalent, establishes a baseline for comparing healthy and diseased groups by quantifying the variation among people, across body habitats and over time. The National Institutes of Health (NIH) sponsored Human Microbiome Project (HMP) has provided an unprecedented opportunity to examine and better define what constitutes the taxonomic core within and across body habitats and individuals through pyrosequencing-based profiling of 16S rRNA gene sequences from oral, skin, distal gut (stool), and vaginal body habitats from over 200 healthy individuals. A two-parameter model is introduced to quantitatively identify the core taxonomic members of each body habitat's microbiota across the healthy cohort. Using only cutoffs for taxonomic ubiquity and abundance, core taxonomic members were identified for each of the 18 body habitats and also for the 4 higher-level body regions. Although many microbes were shared at low abundance, they exhibited a relatively continuous spread in both their abundance and ubiquity, as opposed to a more discretized separation. The numbers of core taxa members in the body regions are comparatively small and stable, reflecting the relatively high, but conserved, interpersonal variability within the cohort. Core sizes increased across the body regions in the order of: vagina, skin, stool, and oral cavity. A number of "minor" oral taxonomic core were also identified by their majority presence across the cohort, but with relatively low and stable abundances. A method for quantifying the difference between two cohorts was introduced and applied to samples collected on a second visit, revealing that over time, the oral, skin, and stool body regions tended to be more transient in their taxonomic structure than the vaginal body region.

  9. Transcriptomic Analyses of Adipocyte Differentiation From Human Mesenchymal Stromal-Cells (MSC).

    PubMed

    Casado-Díaz, Antonio; Anter, Jaouad; Müller, Sören; Winter, Peter; Quesada-Gómez, José Manuel; Dorado, Gabriel

    2017-04-01

    Adipogenesis is a physiological process required for fat-tissue development, mainly involved in regulating the organism energetic-state. Abnormal distribution-changes and dysfunctions in such tissue are associated to different pathologies. Adipocytes are generated from progenitor cells, via a complex differentiating process not yet well understood. Therefore, we investigated differential mRNA and miRNA expression patterns of human mesenchymal stromal-cells (MSC) induced and not induced to differentiate into adipocytes by next (second)-generation sequencing. A total of 2,866 differentially expressed genes (101 encoding miRNA) were identified, with 705 (46 encoding miRNA) being upregulated in adipogenesis. They were related to different pathways, including PPARG, lipid, carbohydrate and energy metabolism, redox, membrane-organelle biosynthesis, and endocrine system. Downregulated genes were related to extracellular matrix and cell migration, proliferation, and differentiation. Analyses of mRNA-miRNA interaction showed that repressed miRNA-encoding genes can act downregulating PPARG-related genes; mostly the PPARG activator (PPARGC1A). Induced miRNA-encoding genes regulate downregulated genes related to TGFB1. These results shed new light to understand adipose-tissue differentiation and physiology, increasing our knowledge about pathologies like obesity, type-2 diabetes and osteoporosis. J. Cell. Physiol. 232: 771-784, 2017. © 2016 Wiley Periodicals, Inc.

  10. Tissue gas and blood analyses of human subjects breathing 80% argon and 20% oxygen

    NASA Technical Reports Server (NTRS)

    Horrigan, D. J.; Wells, C. H.; Guest, M. M.; Hart, G. B.; Goodpasture, J. E.

    1979-01-01

    Eight human volunteers, individually studied in a hyperbaric chamber, breathed: (1) air at 1 ATA; (2) 80% argon and 20% oxygen at 1 ATA for 30 min; (3) air at 1 ATA for 30 min; (4) 100% O2 at 1 ATA for 30 min; (5) air at 1 ATA for 30 min; (6) 100% O2 at 2 ATA for 60 min; and (7) 80% argon and 20% oxygen at 1 ATA for 30 min. Oxygen, carbon dioxide, nitrogen, and argon tensions were measured in muscle and subcutaneous tissue by mass spectroscopic analyses. Venous blood obtained at regular intervals was analyzed for coagulation and fibrinolytic factors. Inert gas narcosis was not observed. After breathing argon for 30 min, muscle argon tensions were almost three times the subcutaneous tensions. Argon wash-in mirrored nitrogen wash-out. Argon wash-in and wash-out had no effect on tissue PO2 or PCO2. Coagulation and fibrinolytic changes usually associated with vascular bubbles were absent.

  11. A Preliminary Study of Biomonitoring for Bisphenol-A in Human Sweat.

    PubMed

    Porucznik, Christina A; Cox, Kyley J; Wilkins, Diana G; Anderson, David J; Bailey, Nicole M; Szczotka, Kathryn M; Stanford, Joseph B

    2015-09-01

    Measurement of human exposure to the endocrine disruptor bisphenol-A (BPA) is hampered by the ubiquitous but transient exposure for most individuals, coupled with a short metabolic half-life which leads to high inter- and intra-individual variability. We investigated the possibility of measuring multiday exposure to BPA in human sweat among volunteer participants with the goal of identifying an exposure assessment method less affected by temporal variability. We recruited 50 participants to wear a sweat collection patch (PharmChek(®)) for 7 days with concurrent collection of daily first-morning urine. Urines and sweat patch extracts were analyzed with quantitative LC-MS-MS using a method we previously validated. In addition, a human volunteer consumed one can of commercially available soup (16 oz, 473 cm(3)) daily for 3 days and collected urine. Sweat patches (n = 2, 1 per arm) were worn for the 3 days of the study. BPA was detected in quality control specimens prepared by fortification of BPA to sweat patches, but was only detected at 5× above average background on three participant patches. Although the highest measured urine BPA concentration was 195 ng/mL for an individual with deliberate exposure, no BPA was detected above background in the corresponding sweat patches. In this preliminary investigation, the use of sweat patches primarily worn on the upper-outer arm did not detect BPA exposures that were documented by urine monitoring. The absence of BPA in sweat patches may be due to several factors, including insufficient quantity of specimen per patch, or extremely low concentrations of BPA in naturally occurring sweat, among others.

  12. Investigation of Human Impacts on Surficial Geomorphology in Mountain Parks Using MASTER Data: Preliminary Results

    NASA Astrophysics Data System (ADS)

    Stefanov, W. L.

    2003-12-01

    The role of humans as active agents of geomorphic change is obvious at large scales (i.e. urbanization), but is also important at smaller temporal and spatial scales due to activities such as hiking and mountain biking. The effect of these activities on soil disturbance, sediment transport, surface hydrology, and vegetation growth are not well understood in arid regions where the majority of projected population growth is expected to occur. The Phoenix, AZ metropolitan area includes several public mountain parks comprising a wide range of bedrock, structure, soils, and geomorphology. The region includes ideal sites for study as there are parks completely surrounded by urbanization (with a high degree of constant use) as well as parks more removed from the metropolitan core that experience less overall human presence. A primarily field-based approach to this study is impractical due to the large total area of investigation (approximately 1000 square kilometers). Use of high spatial and spectral resolution remotely sensed data represents a useful alternative (or complementary) approach. Airborne MODIS/ASTER Simulator (MASTER) data were obtained at a ground resolution of 5 meters/pixel over Phoenix urban and peri-urban mountain parks during April 2003. This sensor acquires data in 50 bands in the visible through mid-infrared wavelengths and captures primary rock-forming mineral (quartz, feldspar, pyroxene, amphibole, etc.); weathering and pedogenetic mineral (clays, oxides, carbonates); and vegetation spectral features useful for compositional analysis. Preliminary results presented here use MASTER spectral data and band ratios to assess the degree of disturbance of surficial soils (as defined by presence of clay minerals and lack of surface crusts/pavements) associated with high- and low-use areas of mountain parks. These initial results are being used to assess the degree to which lithology, structure, vegetation, soil type, and landscape position modulate the

  13. Ultrasonic propulsion of kidney stones: preliminary results of human feasibility study

    PubMed Central

    Bailey, Michael; Cunitz, Bryan; Dunmire, Barbrina; Paun, Marla; Lee, Franklin; Ross, Susan; Lingeman, James; Coburn, Michael; Wessells, Hunter; Sorensen, Mathew; Harper, Jonathan

    2015-01-01

    One in 11 Americans has experienced kidney stones, with a 50% average recurrence rate within 5–10 years. Ultrasonic propulsion (UP) offers a potential method to expel small stones or residual fragments before they become a recurrent problem. Reported here are preliminary findings from the first investigational use of UP in humans. The device uses a Verasonics ultrasound engine and Philips HDI C5-2 probe to generate real-time B-mode imaging and targeted “push” pulses on demand. There are three arms of the study: de novo stones, post-lithotripsy fragments, and the preoperative setting. A pain questionnaire is completed prior to and following the study. Movement is classified based on extent. Patients are followed for 90 days. Ten subjects have been treated to date: three de novo, five post-lithotripsy, and two preoperative. None of the subjects reported pain associated with the treatment or a treatment related adverse event, beyond the normal discomfort of passing a stone. At least one stone was moved in all subjects. Three of five post-lithotripsy subjects passed a single or multiple stones within 1–2 weeks following treatment; one subject passed two (1–2 mm) fragments before leaving clinic. In the pre-operative studies we successfully moved 7 – 8 mm stones. In four subjects, UP revealed multiple stone fragments where the clinical image and initial ultrasound examination indicated a single large stone. PMID:26203347

  14. Volume analysis of heat-induced cracks in human molars: A preliminary study

    PubMed Central

    Sandholzer, Michael A.; Baron, Katharina; Heimel, Patrick; Metscher, Brian D.

    2014-01-01

    Context: Only a few methods have been published dealing with the visualization of heat-induced cracks inside bones and teeth. Aims: As a novel approach this study used nondestructive X-ray microtomography (micro-CT) for volume analysis of heat-induced cracks to observe the reaction of human molars to various levels of thermal stress. Materials and Methods: Eighteen clinically extracted third molars were rehydrated and burned under controlled temperatures (400, 650, and 800°C) using an electric furnace adjusted with a 25°C increase/min. The subsequent high-resolution scans (voxel-size 17.7 μm) were made with a compact micro-CT scanner (SkyScan 1174). In total, 14 scans were automatically segmented with Definiens XD Developer 1.2 and three-dimensional (3D) models were computed with Visage Imaging Amira 5.2.2. The results of the automated segmentation were analyzed with an analysis of variance (ANOVA) and uncorrected post hoc least significant difference (LSD) tests using Statistical Package for Social Sciences (SPSS) 17. A probability level of P < 0.05 was used as an index of statistical significance. Results: A temperature-dependent increase of heat-induced cracks was observed between the three temperature groups (P < 0.05, ANOVA post hoc LSD). In addition, the distributions and shape of the heat-induced changes could be classified using the computed 3D models. Conclusion: The macroscopic heat-induced changes observed in this preliminary study correspond with previous observations of unrestored human teeth, yet the current observations also take into account the entire microscopic 3D expansions of heat-induced cracks within the dental hard tissues. Using the same experimental conditions proposed in the literature, this study confirms previous results, adds new observations, and offers new perspectives in the investigation of forensic evidence. PMID:25125923

  15. Human factors evaluation of remote afterloading brachytherapy. Supporting analyses of human-system interfaces, procedures and practices, training and organizational practices and policies. Volume 3

    SciTech Connect

    Callan, J.R.; Kelly, R.T.; Quinn, M.L.

    1995-07-01

    A human factors project on the use of nuclear by-product material to treat cancer using remotely operated afterloaders was undertaken by the Nuclear Regulatory Commission. The purpose of the project was to identify factors that contribute to human error in the system for remote afterloading brachytherapy (RAB). This report documents the findings from the second, third, fourth, and fifth phases of the project, which involved detailed analyses of four major aspects of the RAB system linked to human error: human-system interfaces; procedures and practices; training practices and policies; and organizational practices and policies, respectively. Findings based on these analyses provided factual and conceptual support for the final phase of this project, which identified factors leading to human error in RAB. The impact of those factors on RAB performance was then evaluated and prioritized in terms of safety significance, and alternative approaches for resolving safety significant problems were identified and evaluated.

  16. Crystallization and preliminary crystallographic studies of a cysteine protease inhibitor from the human nematode parasite Ascaris lumbricoides.

    PubMed

    Liu, Sanling; Dong, Jianmei; Mei, Guoqiang; Liu, Guiyun; Xu, Wei; Su, Zhong; Liu, Jinsong

    2011-02-01

    The cysteine protease inhibitor from Ascaris lumbricoides, a roundworm that lives in the human intestine, may be involved in the suppression of human immune responses. Here, the molecular cloning, protein expression and purification, preliminary crystallization and crystallographic characterization of the cysteine protease inhibitor from A. lumbricoides are reported. The rod-shaped crystal belonged to space group C2, with unit-cell parameters a = 99.40, b = 37.52, c = 62.92 Å, β = 118.26°. The crystal diffracted to 2.1 Å resolution and contained two molecules in the asymmetric unit.

  17. Genetic contributions to regional variability in human brain structure: methods and preliminary results.

    PubMed

    Wright, I C; Sham, P; Murray, R M; Weinberger, D R; Bullmore, E T

    2002-09-01

    limbic areas loaded negatively. Bilateral insula, some frontal regions, and temporal neocortical regions functionally specialized for audition and language loaded strongly on the second PC. We conclude that large samples are required for powerful investigation of genetic effects in imaging data from twins. However, these preliminary re. sults suggest that genetic effects on structure of the human brain are regionally variable and predominantly symmetric in paralimbic structures and lateral temporal cortex.

  18. Comparative Methylome Analyses Identify Epigenetic Regulatory Loci of Human Brain Evolution.

    PubMed

    Mendizabal, Isabel; Shi, Lei; Keller, Thomas E; Konopka, Genevieve; Preuss, Todd M; Hsieh, Tzung-Fu; Hu, Enzhi; Zhang, Zhe; Su, Bing; Yi, Soojin V

    2016-11-01

    How do epigenetic modifications change across species and how do these modifications affect evolution? These are fundamental questions at the forefront of our evolutionary epigenomic understanding. Our previous work investigated human and chimpanzee brain methylomes, but it was limited by the lack of outgroup data which is critical for comparative (epi)genomic studies. Here, we compared whole genome DNA methylation maps from brains of humans, chimpanzees and also rhesus macaques (outgroup) to elucidate DNA methylation changes during human brain evolution. Moreover, we validated that our approach is highly robust by further examining 38 human-specific DMRs using targeted deep genomic and bisulfite sequencing in an independent panel of 37 individuals from five primate species. Our unbiased genome-scan identified human brain differentially methylated regions (DMRs), irrespective of their associations with annotated genes. Remarkably, over half of the newly identified DMRs locate in intergenic regions or gene bodies. Nevertheless, their regulatory potential is on par with those of promoter DMRs. An intriguing observation is that DMRs are enriched in active chromatin loops, suggesting human-specific evolutionary remodeling at a higher-order chromatin structure. These findings indicate that there is substantial reprogramming of epigenomic landscapes during human brain evolution involving noncoding regions.

  19. Comparative Methylome Analyses Identify Epigenetic Regulatory Loci of Human Brain Evolution

    PubMed Central

    Mendizabal, Isabel; Shi, Lei; Keller, Thomas E.; Konopka, Genevieve; Preuss, Todd M.; Hsieh, Tzung-Fu; Hu, Enzhi; Zhang, Zhe; Su, Bing; Yi, Soojin V.

    2016-01-01

    How do epigenetic modifications change across species and how do these modifications affect evolution? These are fundamental questions at the forefront of our evolutionary epigenomic understanding. Our previous work investigated human and chimpanzee brain methylomes, but it was limited by the lack of outgroup data which is critical for comparative (epi)genomic studies. Here, we compared whole genome DNA methylation maps from brains of humans, chimpanzees and also rhesus macaques (outgroup) to elucidate DNA methylation changes during human brain evolution. Moreover, we validated that our approach is highly robust by further examining 38 human-specific DMRs using targeted deep genomic and bisulfite sequencing in an independent panel of 37 individuals from five primate species. Our unbiased genome-scan identified human brain differentially methylated regions (DMRs), irrespective of their associations with annotated genes. Remarkably, over half of the newly identified DMRs locate in intergenic regions or gene bodies. Nevertheless, their regulatory potential is on par with those of promoter DMRs. An intriguing observation is that DMRs are enriched in active chromatin loops, suggesting human-specific evolutionary remodeling at a higher-order chromatin structure. These findings indicate that there is substantial reprogramming of epigenomic landscapes during human brain evolution involving noncoding regions. PMID:27563052

  20. Functional Analyses of Transcription Factor Binding Sites that Differ between Present-Day and Archaic Humans

    PubMed Central

    Weyer, Sven; Pääbo, Svante

    2016-01-01

    We analyze 25 previously identified transcription factor binding sites that carry DNA sequence changes that are present in all or nearly all present-day humans, yet occur in the ancestral state in Neandertals and Denisovans, the closest evolutionary relatives of humans. When the ancestral and derived forms of the transcription factor binding sites are tested using reporter constructs in 3 neuronal cell lines, the activity of 12 of the derived versions of transcription factor binding sites differ from the respective ancestral variants. This suggests that the majority of this class of evolutionary differences between modern humans and Neandertals may affect gene expression in at least some tissue or cell type. PMID:26454764

  1. Experiments on Analysing Voice Production: Excised (Human, Animal) and In Vivo (Animal) Approaches.

    PubMed

    Döllinger, Michael; Kobler, James; Berry, David A; Mehta, Daryush D; Luegmair, Georg; Bohr, Christopher

    Experiments on human and on animal excised specimens as well as in vivo animal preparations are so far the most realistic approaches to simulate the in vivo process of human phonation. These experiments do not have the disadvantage of limited space within the neck and enable studies of the actual organ necessary for phonation, i.e., the larynx. The studies additionally allow the analysis of flow, vocal fold dynamics, and resulting acoustics in relation to well-defined laryngeal alterations.

  2. U-series and radiocarbon analyses of human and faunal remains from Wajak, Indonesia.

    PubMed

    Storm, Paul; Wood, Rachel; Stringer, Chris; Bartsiokas, Antonis; de Vos, John; Aubert, Maxime; Kinsley, Les; Grün, Rainer

    2013-05-01

    Laser ablation U-series dating results on human and faunal bone fragments from Wajak, Indonesia, indicate a minimum age of between 37.4 and 28.5 ka (thousands of years ago) for the whole assemblage. These are significantly older than previously published radiocarbon estimates on bone carbonate, which suggested a Holocene age for a human bone fragment and a late Pleistocene age for a faunal bone. The analysis of the organic components in the faunal material show severe degradation and a positive δ(13)C ratio indicate a high degree of secondary carbonatisation. This may explain why the thermal release method used for the original age assessments yielded such young ages. While the older U-series ages are not in contradiction with the morphology of the Wajak human fossils or Javanese biostratigraphy, they will require a reassessment of the evolutionary relationships of modern human remains in Southeast Asia and Oceania. It can be expected that systematic direct dating of human fossils from this area will lead to further revisions of our understanding of modern human evolution.

  3. Molecular analyses of in vivo hprt mutations in human T-lymphocytes: IV. Studies in newborns

    SciTech Connect

    McGinniss, M.J.; Nicklas, J.A.; Albertini, R.J. )

    1989-01-01

    In order to characterize in vivo gene mutations that occur during fetal development, molecular analyses were undertaken of mutant 6-thioguanine resistant T-lymphocytes isolated from placental cord blood samples of 13 normal male newborns. These mutant T-cells were studied to define hypoxanthine-guanine phosphoribosyltransferase (hprt) gene structural alterations and to determine T-cell receptor (TCR) gene rearrangement patterns. Structural hprt alterations, as shown by Southern blot analyses, occurred in 85% of these mutant clones. These alterations consisted mostly of deletion of exons 2 and 3. These findings contrast with the 10-20% of gross structural alterations occurring randomly across the entire gene previously reported for T-cell mutants isolated from normal young adults. Iterative analyses of hprt structural alterations and TCR gene rearrangement patterns show that approximately one-third of the newborn derived mutants may have originated as pre- or intrathymic hprt mutations. This too contrasts with previous findings in adults where the background in vivo hprt mutations appeared to originate in postthymic T-lymphocytes.

  4. Preliminary genetic imaging study of the association between estrogen receptor-α gene polymorphisms and harsh human maternal parenting.

    PubMed

    Lahey, Benjamin B; Michalska, Kalina J; Liu, Chunyu; Chen, Qi; Hipwell, Alison E; Chronis-Tuscano, Andrea; Waldman, Irwin D; Decety, Jean

    2012-09-06

    A failure of neural changes initiated by the estrogen surge in late pregnancy to reverse the valence of infant stimuli from aversive to rewarding is associated with dysfunctional maternal behavior in nonhuman mammals. Estrogen receptor-α plays the crucial role in mediating these neural effects of estrogen priming. This preliminary study examines associations between estrogen receptor-α gene polymorphisms and human maternal behavior. Two polymorphisms were associated with human negative maternal parenting. Furthermore, hemodynamic responses in functional magnetic resonance imaging to child stimuli in neural regions associated with social cognition fully mediated the association between genetic variation and negative parenting. This suggests testable hypotheses regarding a biological pathway between genetic variants and dysfunctional human maternal parenting.

  5. Human Infections with Spirometra decipiens Plerocercoids Identified by Morphologic and Genetic Analyses in Korea.

    PubMed

    Jeon, Hyeong-Kyu; Park, Hansol; Lee, Dongmin; Choe, Seongjun; Kim, Kyu-Heon; Huh, Sun; Sohn, Woon-Mok; Chai, Jong-Yil; Eom, Keeseon S

    2015-06-01

    Tapeworms of the genus Spirometra are pseudophyllidean cestodes endemic in Korea. At present, it is unclear which Spirometra species are responsible for causing human infections, and little information is available on the epidemiological profiles of Spirometra species infecting humans in Korea. Between 1979 and 2009, a total of 50 spargana from human patients and 2 adult specimens obtained from experimentally infected carnivorous animals were analyzed according to genetic and taxonomic criteria and classified as Spirometra erinaceieuropaei or Spirometra decipiens depending on the morphology. Morphologically, S. erinaceieuropaei and S. decipiens are different in that the spirally coiled uterus in S. erinaceieuropaei has 5-7 complete coils, while in S. decipiens it has only 4.5 coils. In addition, there is a 9.3% (146/1,566) sequence different between S. erinaceieuropaei and S. decipiens in the cox1 gene. Partial cox1 sequences (390 bp) from 35 Korean isolates showed 99.4% (388/390) similarity with the reference sequence of S. erinaceieuropaei from Korea (G1724; GenBank KJ599680) and an additional 15 Korean isolates revealed 99.2% (387/390) similarity with the reference sequences of S. decipiens from Korea (G1657; GenBank KJ599679). Based on morphologic and molecular databases, the estimated population ratio of S. erinaceieuropaei to S. decipiens was 35: 15. Our results indicate that both S. erinaceieuropaei and S. decipiens found in Korea infect humans, with S. erinaceieuropaei being 2 times more prevalent than S. decipiens. This study is the first to report human sparganosis caused by S. decipiens in humans in Korea.

  6. Evolution of the human hand: approaches to acquiring, analysing and interpreting the anatomical evidence

    PubMed Central

    MARZKE, MARY W.; MARZKE, R. F.

    2000-01-01

    The discovery of fossil hand bones from an early human ancestor at Olduvai Gorge in 1960, at the same level as primitive stone tools, generated a debate about the role of tools in the evolution of the human hand that has raged to the present day. Could the Olduvai hand have made the tools? Did the human hand evolve as an adaptation to tool making and tool use? The debate has been fueled by anatomical studies comparing living and fossil human and nonhuman primate hands, and by experimental observations. These have assessed the relative abilities of apes and humans to manufacture the Oldowan tools, but consensus has been hampered by disagreements about how to translate experimental data from living species into quantitative models for predicting the performance of fossil hands. Such models are now beginning to take shape as new techniques are applied to the capture, management and analysis of data on kinetic and kinematic variables ranging from hand joint structure, muscle mechanics, and the distribution and density of bone to joint movements and muscle recruitment during manipulative behaviour. The systematic comparative studies are highlighting a functional complex of features in the human hand facilitating a distinctive repertoire of grips that are apparently more effective for stone tool making than grips characterising various nonhuman primate species. The new techniques are identifying skeletal variables whose form may provide clues to the potential of fossil hominid hands for one-handed firm precision grips and fine precision manoeuvering movements, both of which are essential for habitual and effective tool making and tool use. PMID:10999274

  7. Preference and consequences: A preliminary look at whether preference impacts oral processing in non-human primates.

    PubMed

    Vinyard, Christopher J; Thompson, Cynthia L; Doherty, Alison; Robl, Nicholas

    2016-09-01

    Non-human primates demonstrate food preferences much like humans. We have little insight, however, into how those preferences impact oral processing in primates. To begin describing this relationship, we conducted a preliminary analysis measuring food preference in two tufted capuchins (Cebus apella) and comparing ranked preference to physiological variables during chewing of these foods. Food preference was assessed for each monkey across 12 foods, including monkey biscuits and 11 foods consumed by humans (e.g., various fruits and nuts). Animals chose from randomized pairs of foods to generate a ranked scale across the 12 foods. Contemporaneous with preference testing, electromyographic (EMG) activity was measured for the jaw-closing muscles to assess oral physiology during chewing of these foods. As expected, these capuchins exhibited clear preferences among these 12 foods. Based on their preferences, we identified sets of preferred and non-preferred brittle (i.e., almond versus monkey chow) and ductile (i.e., dates and prunes versus apricots) foods for physiological comparisons that broadly control variation in food mechanical properties (FMPs). As expected, oral physiology varied with FMPs in each animal. Within brittle and ductile groupings, we observed several significant differences in chewing cycle length and relative muscle activation levels that are likely related to food preference. These differences tended to be complex and individual specific. The two capuchins chewed non-preferred apricots significantly faster than preferred dates and prunes. Effect sizes for preference were smaller than those for FMPs, supporting the previous focus on FMPs in primate dietary research. Although preliminary, these results suggest that food preference may influence oral physiology in non-human primates. The prospect that this relationship exists in monkeys raises the possibility that a link between food preference and oral processing in humans may be based on shared

  8. A preliminary 3D computed tomography study of the human maxillary sinus and nasal cavity.

    PubMed

    Butaric, Lauren N; McCarthy, Robert C; Broadfield, Douglas C

    2010-11-01

    Despite centuries of investigation, the function of the maxillary sinus (MS) and underlying patterns governing its form remain elusive. In this study, we articulate a methodology for collecting volumetric data for the MS and nasal cavity (NC) from computed tomography (CT) scans and report details for a small sample of 39 dried human crania of known ecogeographic provenience useful for assessing variation in MS size and shape. We use scaling analyses to preliminarily test the hypothesis that volumes of the nasal cavity (NCV) and maxillary sinus (MSV) are inversely correlated such that the NC covaries with size of the face, whereas the MS "fills in" the leftover space [proposed by Shea: Am J Phys Anthropol 47 (1977):289-300]. Against expectation, MSV is not significantly correlated with NCV or any cranial size variable. NCV, on the other hand, scales isometrically with facial size. The results of this pilot study suggest that NCV covaries with facial size, but that the MS does not simply fill in the leftover space in the face. The role, if any, of the MSs in midfacial function and architecture remains unclear. Larger sample sizes, additional environmental variables, and assessment of MS and NC shape are necessary to resolve this issue.

  9. Health and Human Services. Occupational Analyses. Worker Task Lists and Supplementary Information for Selected Occupations.

    ERIC Educational Resources Information Center

    Henrico County Public Schools, Glen Allen, VA. Virginia Vocational Curriculum and Resource Center.

    This publication contains worker task lists and supplementary information for eight occupations in the health and human services cluster: (1) criminal justice; (2) protective services; (3) dental assistant; (4) dental hygienist; (5) diagnostic medical sonographer; (6) medical office assistant; (7) fire medic; and (8) parks and recreation manager.…

  10. Human Rights in Social Science Textbooks: Cross-National Analyses, 1970-2008

    ERIC Educational Resources Information Center

    Meyer, John W.; Bromley, Patricia; Ramirez, Francisco O.

    2010-01-01

    In reaction to the disasters of the first half the 20th century and World War II, a dramatic world movement arose emphasizing the human rights of persons in global society. The contrast--celebrated in international treaties, intergovernmental and nongovernmental organizations, and much cultural discourse--was with narrower world emphases on the…

  11. UNCERTAINTY AND SENSITIVITY ANALYSES FOR INTEGRATED HUMAN HEALTH AND ECOLOGICAL RISK ASSESSMENT OF HAZARDOUS WASTE DISPOSAL

    EPA Science Inventory

    While there is a high potential for exposure of humans and ecosystems to chemicals released from hazardous waste sites, the degree to which this potential is realized is often uncertain. Conceptually divided among parameter, model, and modeler uncertainties imparted during simula...

  12. Integrated Genomic and Network-Based Analyses of Complex Diseases and Human Disease Network.

    PubMed

    Al-Harazi, Olfat; Al Insaif, Sadiq; Al-Ajlan, Monirah A; Kaya, Namik; Dzimiri, Nduna; Colak, Dilek

    2016-06-20

    A disease phenotype generally reflects various pathobiological processes that interact in a complex network. The highly interconnected nature of the human protein interaction network (interactome) indicates that, at the molecular level, it is difficult to consider diseases as being independent of one another. Recently, genome-wide molecular measurements, data mining and bioinformatics approaches have provided the means to explore human diseases from a molecular basis. The exploration of diseases and a system of disease relationships based on the integration of genome-wide molecular data with the human interactome could offer a powerful perspective for understanding the molecular architecture of diseases. Recently, subnetwork markers have proven to be more robust and reliable than individual biomarker genes selected based on gene expression profiles alone, and achieve higher accuracy in disease classification. We have applied one of these methodologies to idiopathic dilated cardiomyopathy (IDCM) data that we have generated using a microarray and identified significant subnetworks associated with the disease. In this paper, we review the recent endeavours in this direction, and summarize the existing methodologies and computational tools for network-based analysis of complex diseases and molecular relationships among apparently different disorders and human disease network. We also discuss the future research trends and topics of this promising field.

  13. Preliminary studies of fluorescence image-guided photothermal therapy of human oesophageal adenocarcinoma in vivo using multifunctional gold nanorods

    NASA Astrophysics Data System (ADS)

    Nabavi, Elham; Singh, Mohan; Zhou, Yu; Gallina, Maria Elena; Zhao, Hailin; Ma, Daqing; Cass, Anthony; Hanna, George; Elson, Daniel S.

    2016-03-01

    We present a preliminary in vivo study of fluorescence imaging and photothermal therapy (PTT) of human oesophageal adenocarcinoma using multi-functionalised gold nanorods (GNRs). After establishing tumour xenograft in mouse functionalised GNRs were administrated intravenously (IV). Fluorescence imaging was performed to detect the tumour area. The intensity of the fluorescence signal varied significantly across the tumour site and surrounding tissues. PTT was then performed using a 808 nm continuous wave diode laser to irradiate the tumour for 3 minutes, inducing a temperature rise of ~44°C, which photothermally ablated the tumour.

  14. Human-arm-and-hand-dynamic model with variability analyses for a stylus-based haptic interface.

    PubMed

    Fu, Michael J; Cavuşoğlu, M Cenk

    2012-12-01

    Haptic interface research benefits from accurate human arm models for control and system design. The literature contains many human arm dynamic models but lacks detailed variability analyses. Without accurate measurements, variability is modeled in a very conservative manner, leading to less than optimal controller and system designs. This paper not only presents models for human arm dynamics but also develops inter- and intrasubject variability models for a stylus-based haptic device. Data from 15 human subjects (nine male, six female, ages 20-32) were collected using a Phantom Premium 1.5a haptic device for system identification. In this paper, grip-force-dependent models were identified for 1-3-N grip forces in the three spatial axes. Also, variability due to human subjects and grip-force variation were modeled as both structured and unstructured uncertainties. For both forms of variability, the maximum variation, 95 %, and 67 % confidence interval limits were examined. All models were in the frequency domain with force as input and position as output. The identified models enable precise controllers targeted to a subset of possible human operator dynamics.

  15. Functional Analyses of Human DNA Repair Proteins Important for Aging and Genomic Stability Using Yeast Genetics

    PubMed Central

    Aggarwal, Monika; Brosh, Robert M.

    2012-01-01

    Model systems have been extremely useful for studying various theories of aging. Studies of yeast have been particularly helpful to explore the molecular mechanisms and pathways that affect aging at the cellular level in the simple eukaryote. Although genetic analysis has been useful to interrogate the aging process, there has been both interest and debate over how functionally conserved the mechanisms of aging are between yeast and higher eukaryotes, especially mammalian cells. One area of interest has been the importance of genomic stability for age-related processes, and the potential conservation of proteins and pathways between yeast and human. Translational genetics have been employed to examine the functional roles of mammalian proteins using yeast as a pliable model system. In the current review recent advancements made in this area are discussed, highlighting work which shows that the cellular functions of human proteins in DNA repair and maintenance of genomic stability can be elucidated by genetic rescue experiments performed in yeast. PMID:22349084

  16. Preliminary results on the tectonic activity of the Ovacık Fault (Malatya-Ovacık Fault Zone, Turkey): Implications of the morphometric analyses

    NASA Astrophysics Data System (ADS)

    Yazıcı, Müge; Zabci, Cengiz; Sançar, Taylan; Sunal, Gürsel; Natalin, Boris A.

    2016-04-01

    The Anatolian 'plate' is being extruded westward relative to the Eurasia along two major tectonic structures, the North Anatolian and the East Anatolian shear zones, respectively making its northern and eastern boundaries. Although the main deformation is localized along these two structures, there is remarkable intra-plate deformation within Anatolia, especially which are characterized by NE-striking sinistral and NW-striking dextral strike-slip faults (Şengör et al. 1985). The Malatya-Ovacık Fault Zone (MOFZ) and its northeastern member, the Ovacık Fault (OF), is a one of the NE-striking sinistral strike slip faults in the central 'ova' neotectonic province of Anatolia, located close to its eastern boundary. Although this fault zone is claimed to be an inactive structure in some studies, the recent GPS measurements (Aktuǧ et al., 2013) and microseismic activity (AFAD, 2013) strongly suggest the opposite. In order to understand rates and patterns of vertical ground motions along the OF, we studied the certain morphometric analyses such as hypsometric curves and integrals, longitudinal channel profiles, and asymmetry of drainage basins. The Karasu (Euphrates) and Munzur rivers form the main drainage systems of the study area. We extracted all drainage network from SRTM-based Digital Elevation Model with 30 m ground pixel resolution and totally identified 40 sub-drainage basins, which are inhomogeneously distributed to the north and to the south of the OF. Most of these basins show strong asymmetry, which are mainly tilted to SW. The asymmetry relatively decreases from NE to SW in general. The only exception is at the margins of the Ovacık Basin (OB), where almost the highest asymmetry values were calculated. On the other hand, the characteristics of hypsometric curves and the calculated hypsometric integrals do not show the similar systematic spatial pattern. The hypsometric curves with convex-shaped geometry, naturally indicating relatively young morphology

  17. Computational analyses of CO-rebreathing methods for estimating haemoglobin mass in humans.

    PubMed

    Chada, Kinnera E; Bruce, Eugene N

    2012-01-01

    Measurement of haemoglobin mass (M(Hb)) is used to quantify alterations in oxygen delivery during exercise training or acclimatization to altitude. Uptake of carbon monoxide by haemoglobin is the basis of the common non-radioactive methods to determine M(Hb) in humans. This study used a validated mathematical model to simulate CO uptake during rebreathing protocols and to determine sources of errors in estimation of M(Hb). Our previously published model was validated using experimentally measured carboxyhaemoglobin levels (%HbCO) from arterial, capillary and venous blood sites of human subjects during CO-rebreathing protocols. This model was then used to simulate various CO-rebreathing protocols in 24 human subjects with known M(Hb). Using variables generated by the model, M(Hb) was estimated on the basis of assumptions typically made for calculating the volume of CO bound to myoglobin, the volume of CO exhaled and the volume of CO in the rebreathing system. It was found that inaccurate estimation of the volume of CO bound to myoglobin was the major source of error in determination of M(Hb). Additionally, the size of the error was found to depend on the site of blood sampling because of differences in %HbCO. Regression equations were developed to improve the estimation of volume of CO bound to myoglobin, and a new protocol that is less dependent on the site of blood sampling is proposed.

  18. Multidimensional Genome-wide Analyses Show Accurate FVIII Integration by ZFN in Primary Human Cells

    PubMed Central

    Sivalingam, Jaichandran; Kenanov, Dimitar; Han, Hao; Nirmal, Ajit Johnson; Ng, Wai Har; Lee, Sze Sing; Masilamani, Jeyakumar; Phan, Toan Thang; Maurer-Stroh, Sebastian; Kon, Oi Lian

    2016-01-01

    Costly coagulation factor VIII (FVIII) replacement therapy is a barrier to optimal clinical management of hemophilia A. Therapy using FVIII-secreting autologous primary cells is potentially efficacious and more affordable. Zinc finger nucleases (ZFN) mediate transgene integration into the AAVS1 locus but comprehensive evaluation of off-target genome effects is currently lacking. In light of serious adverse effects in clinical trials which employed genome-integrating viral vectors, this study evaluated potential genotoxicity of ZFN-mediated transgenesis using different techniques. We employed deep sequencing of predicted off-target sites, copy number analysis, whole-genome sequencing, and RNA-seq in primary human umbilical cord-lining epithelial cells (CLECs) with AAVS1 ZFN-mediated FVIII transgene integration. We combined molecular features to enhance the accuracy and activity of ZFN-mediated transgenesis. Our data showed a low frequency of ZFN-associated indels, no detectable off-target transgene integrations or chromosomal rearrangements. ZFN-modified CLECs had very few dysregulated transcripts and no evidence of activated oncogenic pathways. We also showed AAVS1 ZFN activity and durable FVIII transgene secretion in primary human dermal fibroblasts, bone marrow- and adipose tissue-derived stromal cells. Our study suggests that, with close attention to the molecular design of genome-modifying constructs, AAVS1 ZFN-mediated FVIII integration in several primary human cell types may be safe and efficacious. PMID:26689265

  19. Molecular and immunochemical analyses of RB1 and cyclin D1 in human ductal pancreatic carcinomas and cell lines.

    PubMed

    Huang, L; Lang, D; Geradts, J; Obara, T; Klein-Szanto, A J; Lynch, H T; Ruggeri, B A

    1996-02-01

    Somatic mutations in the retinoblastoma-1 gene (RB1) and loss of RB1 protein function have been implicated in a number of human malignancies, but the role of RB1 gene and protein abnormalities in ductal pancreatic cancer (DPCA) is virtually unknown. We therefore analyzed expression of the RB1 protein immunohistochemically and/or by western blotting in a total of 54 sporadic and eight familial cases of archival and frozen DPCA and in 18 pancreatic carcinoma cell lines by using the antibodies RB-WL-1, 84-B3-1, and PMG3-245. Mutations in the RB1 promotor region and exons 13-21 of the RB1 gene were likewise examined by single-strand conformation polymorphism (SSCP) analyses and DNA sequencing of genomic DNA from 30 microdissected primary pancreatic tumors and the pancreatic carcinoma cell lines. Moreover, amplification and expression of a major regulatory component of RB1 function, cyclin D1, were assessed by southern and immunohistochemical analyses, respectively. The DPCAs were heterogeneous in both the intensity of RB1 nuclear staining and the percentage of immunoreactive cells. The tumors often had areas where RB1 staining was weak or absent adjacent to normal pancreatic tissue; however, only two of 32 archival cases and one of 30 frozen cases of DPCA completely lacked RB1 nuclear staining. Immunohistochemical and western blot analyses of 18 pancreatic carcinoma cell lines demonstrated the absence of RB1 expression in only two cell lines, Capan-1 and QGP-1. Analyses of the RB1 gene and promotor region by SSCP and DNA sequencing largely confirmed the immunochemical findings. Three of 30 primary carcinomas had abnormalities revealed by SSCP analyses. In one case a single base-pair deletion was confirmed in exon 18 and resulted in premature termination and the absence of detectable RB1 protein. A second case had TAC-->TTC missense mutation in exon 13. The third primary carcinoma could not be reliably sequenced because it had a low percentage of epithelial cells. The

  20. Genomic and in silico analyses of CRBN gene and thalidomide embryopathy in humans.

    PubMed

    Vianna, Fernanda Sales Luiz; Kowalski, Thayne Woycinck; Tovo-Rodrigues, Luciana; Tagliani-Ribeiro, Alice; Godoy, Bibiane Armiliato; Fraga, Lucas Rosa; Sanseverino, Maria Teresa Vieira; Hutz, Mara Helena; Schuler-Faccini, Lavínia

    2016-12-01

    Thalidomide causes Thalidomide Embryopathy (TE), but is largely used to treat several conditions. Investigations with Cereblon, a thalidomide target protein encoded by CRBN gene, have helped to understand thalidomide therapeutic and teratogenic properties. We sequenced CRBN-thalidomide binding region in 38 TE individuals and 136 Brazilians without congenital anomalies, and performed in silico analyses. Eight variants were identified, seven intronic and one in 3'UTR. TE individuals had rare variants in higher frequency than the non-affected group (p=0.04). The genotype rs1620675 CC was related to neurological anomalies in TE individuals (p=0.004). Bioinformatics analysis suggested this genotype leads to potential alterations in splicing sites and binding to transcription factors. Comparison of the Cereblon-thalidomide binding domains in mammals demonstrated that CRBN is highly conserved across species. All the variants require evaluation in functional assays in order to understand their role in Cereblon-thalidomide binding and complex interactions that lead to TE.

  1. Assessing Human Diet and Movement in the Tongan Maritime Chiefdom Using Isotopic Analyses

    PubMed Central

    Stantis, Christina; Kinaston, Rebecca L.; Richards, Michael P.; Davidson, Janet M.; Buckley, Hallie R.

    2015-01-01

    The rise of stratified societies fundamentally influences the interactions between status, movement, and food. Using isotopic analyses, we assess differences in diet and mobility of individuals excavated from two burial mounds located at the `Atele burial site on Tongatapu, the main island of the Kingdom of Tonga (c. 500 - 150 BP). The first burial mound (To-At-1) was classified by some archaeologists as a commoner’s mound while the second burial mound (To-At-2) was possibly used for interment of the chiefly class. In this study, stable isotope analyses of diet (δ13C, δ15N, and δ34S; n = 41) are used to asses paleodiet and 87Sr/86Sr ratios (n = 30) are analyzed to investigate individual mobility to test whether sex and social status affected these aspects of life. Our results show significant differences in diet between burial mounds and sexes. Those interred in To-At-2 displayed lower δ13C values, indicating they ate relatively more terrestrial plants (likely starchy vegetable staples) compared with To-At-1 individuals. Females displayed significantly lower δ15N values compared with males within the entire assemblage. No differences in δ34S values were observed between sexes or burial mound but it is possible that sea spray or volcanism may have affected these values. One individual displayed the strontium isotopic composition representative of a nonlocal immigrant (outside 2SD of the mean). This suggests the hegemonic control over interisland travel, may have prevented long-term access to the island by non-Tongans exemplifying the political and spiritual importance of the island of Tongatapu in the maritime chiefdom. PMID:25822619

  2. Specific binding of /sup 125/I-labeled human chorionic gonadotropin to gonadal tissue: comparison of limited-point saturation analyses to Scatchard analyses for determining binding capacities and factors affecting estimates of binding capacity

    SciTech Connect

    Spicer, L.J.; Ireland, J.J.

    1986-07-01

    Experiments were conducted to compare gonadotropin binding capacity calculated from limited-point saturation analyses to those obtained from Scatchard analyses, and to test the effects of membrane purity and source of gonadotropin receptors on determining the maximum percentage of radioiodinated hormone bound to receptors (maximum bindability). One- to four-point saturation analyses gave results comparable to results by Scatchard analyses when examining relative binding capacities of receptors. Crude testicular homogenates had lower estimates of maximum bindability of /sup 125/I-labeled human chorionic gonadotropin than more purified gonadotropin receptor preparations. Under similar preparation techniques, some gonadotropin receptor sources exhibited low maximum bindability.

  3. Experiments on Analysing Voice Production: Excised (Human, Animal) and In Vivo (Animal) Approaches

    PubMed Central

    Döllinger, Michael; Kobler, James; Berry, David A.; Mehta, Daryush D.; Luegmair, Georg; Bohr, Christopher

    2015-01-01

    Experiments on human and on animal excised specimens as well as in vivo animal preparations are so far the most realistic approaches to simulate the in vivo process of human phonation. These experiments do not have the disadvantage of limited space within the neck and enable studies of the actual organ necessary for phonation, i.e., the larynx. The studies additionally allow the analysis of flow, vocal fold dynamics, and resulting acoustics in relation to well-defined laryngeal alterations. Purpose of Review This paper provides an overview of the applications and usefulness of excised (human/animal) specimen and in vivo animal experiments in voice research. These experiments have enabled visualization and analysis of dehydration effects, vocal fold scarring, bifurcation and chaotic vibrations, three-dimensional vibrations, aerodynamic effects, and mucosal wave propagation along the medial surface. Quantitative data will be shown to give an overview of measured laryngeal parameter values. As yet, a full understanding of all existing interactions in voice production has not been achieved, and thus, where possible, we try to indicate areas needing further study. Recent Findings A further motivation behind this review is to highlight recent findings and technologies related to the study of vocal fold dynamics and its applications. For example, studies of interactions between vocal tract airflow and generation of acoustics have recently shown that airflow superior to the glottis is governed by not only vocal fold dynamics but also by subglottal and supraglottal structures. In addition, promising new methods to investigate kinematics and dynamics have been reported recently, including dynamic optical coherence tomography, X-ray stroboscopy and three-dimensional reconstruction with laser projection systems. Finally, we touch on the relevance of vocal fold dynamics to clinical laryngology and to clinically-oriented research. PMID:26581597

  4. Vasoactive intestinal polypeptide immunoreactivity in the human cerebellum: qualitative and quantitative analyses

    PubMed Central

    Benagiano, Vincenzo; Flace, Paolo; Lorusso, Loredana; Rizzi, Anna; Bosco, Lorenzo; Cagiano, Raffaele; Ambrosi, Glauco

    2009-01-01

    Although autoradiographic, reverse transcription-polymerase chain reaction and immunohistochemical studies have demonstrated receptors for vasoactive intestinal polypeptide (VIP) in the cerebellum of various species, immunohistochemistry has never shown immunoreactivity for VIP within cerebellar neuronal bodies and processes. The present study aimed to ascertain whether VIP immunoreactivity really does exist in the human cerebellum by making a systematic analysis of samples removed post-mortem from all of the cerebellar lobes. The study was carried out using light microscopy immunohistochemical techniques based on a set of four different antibodies (three polyclonal and one monoclonal) against VIP, carefully selected on the basis of control tests performed on human colon. All of the antibodies used showed VIP-immunoreactive neuronal bodies and processes distributed in the cerebellar cortex and subjacent white matter of all of the cerebellum lobes, having similar qualitative patterns of distribution. Immunoreactive neurons included subpopulations of the main neuron types of the cortex. Statistical analysis of the quantitative data on the VIP immunoreactivity revealed by the different antibodies in the different cerebellar lobes did not demonstrate any significant differences. In conclusion, using four different anti-VIP antibodies, the first evidence of VIP immunoreactivity is herein supplied in the human post-mortem cerebellum, with similar qualitative/quantitative patterns of distribution among the different cerebellum lobes. Owing to the function performed by VIP as a neurotransmitter/neuromodulator, it is a candidate for a role in intrinsic and extrinsic (projective) circuits of the cerebellum, in agreement with previous demonstrations of receptors for VIP in the cerebellar cortex and nuclei. As VIP signalling pathways are implicated in the regulation of cognitive and psychic functions, cerebral blood flow and metabolism, processes of histomorphogenesis

  5. Isolation and transcriptome analyses of human erythroid progenitors: BFU-E and CFU-E.

    PubMed

    Li, Jie; Hale, John; Bhagia, Pooja; Xue, Fumin; Chen, Lixiang; Jaffray, Julie; Yan, Hongxia; Lane, Joseph; Gallagher, Patrick G; Mohandas, Narla; Liu, Jing; An, Xiuli

    2014-12-04

    Burst-forming unit-erythroid (BFU-E) and colony-forming unit-erythroid (CFU-E) cells are erythroid progenitors traditionally defined by colony assays. We developed a flow cytometry-based strategy for isolating human BFU-E and CFU-E cells based on the changes in expression of cell surface markers during in vitro erythroid cell culture. BFU-E and CFU-E are characterized by CD45(+)GPA(-)IL-3R(-)CD34(+)CD36(-)CD71(low) and CD45(+)GPA(-)IL-3R(-)CD34(-)CD36(+)CD71(high) phenotypes, respectively. Colony assays validated phenotypic assignment giving rise to BFU-E and CFU-E colonies, both at a purity of ∼90%. The BFU-E colony forming ability of CD45(+)GPA(-)IL-3R(-)CD34(+)CD36(-)CD71(low) cells required stem cell factor and erythropoietin, while the CFU-E colony forming ability of CD45(+)GPA(-)IL-3R(-)CD34(-)CD36(+)CD71(high) cells required only erythropoietin. Bioinformatic analysis of the RNA-sequencing data revealed unique transcriptomes at each differentiation stage. The sorting strategy was validated in uncultured primary cells isolated from bone marrow, cord blood, and peripheral blood, indicating that marker expression is not an artifact of in vitro cell culture, but represents an in vivo characteristic of erythroid progenitor populations. The ability to isolate highly pure human BFU-E and CFU-E progenitors will enable detailed cellular and molecular characterization of these distinct progenitor populations and define their contribution to disordered erythropoiesis in inherited and acquired hematologic disease. Our data provides an important resource for future studies of human erythropoiesis.

  6. Comparative Proteome Analyses of Human Plasma Following in vivo Lipopolysaccharide Administration Using Multidimensional Separations Coupled with Tandem Mass Spectrometry

    PubMed Central

    Qian, Wei-Jun; Jacobs, Jon M.; Camp, David G.; Monroe, Matthew E.; Moore, Ronald J.; Gritsenko, Marina A.; Calvano, Steve E.; Lowry, Stephen F.; Xiao, Wenzhong; Moldawer, Lyle L.; Davis, Ronald W.; Tompkins, Ronald G.; Smith, Richard D.

    2007-01-01

    There is significant interest in characterization of the human plasma proteome due to its potential for providing biomarkers applicable to clinical diagnosis and treatment and for gaining a better understanding of human diseases. We describe here a strategy for the comparative proteome analyses of human plasma, which is applicable to biomarker identifications for various disease states. Multidimensional liquid chromatography-mass spectrometry has been applied to make comparative proteome analyses of plasma samples from an individual prior to and 9 h after lipopolysaccharide (LPS) administration. Peptide peak areas and the number of peptide identifications for each protein were used to evaluate the reproducibility of LC-MS/MS and to compare relative changes in protein concentration between the samples following LPS treatment. A total of 804 distinct plasma proteins (not including immunoglobulins) were confidently identified with 32 proteins observed to be significantly increased in concentration following LPS administration, including several known inflammatory response or acute-phase mediators such as C-reactive protein, serum amyloid A and A2, LPS-binding protein, LPS-responsive and beige-like anchor protein, hepatocyte growth factor activator, and von Willebrand factor, and thus constituting potential biomarkers for inflammatory response. PMID:15627965

  7. Comparative Genomic Analyses of the Human NPHP1 Locus Reveal Complex Genomic Architecture and Its Regional Evolution in Primates

    PubMed Central

    Yuan, Bo; Liu, Pengfei; Gupta, Aditya; Beck, Christine R.; Tejomurtula, Anusha; Campbell, Ian M.; Gambin, Tomasz; Simmons, Alexandra D.; Withers, Marjorie A.; Harris, R. Alan; Rogers, Jeffrey; Schwartz, David C.; Lupski, James R.

    2015-01-01

    Many loci in the human genome harbor complex genomic structures that can result in susceptibility to genomic rearrangements leading to various genomic disorders. Nephronophthisis 1 (NPHP1, MIM# 256100) is an autosomal recessive disorder that can be caused by defects of NPHP1; the gene maps within the human 2q13 region where low copy repeats (LCRs) are abundant. Loss of function of NPHP1 is responsible for approximately 85% of the NPHP1 cases—about 80% of such individuals carry a large recurrent homozygous NPHP1 deletion that occurs via nonallelic homologous recombination (NAHR) between two flanking directly oriented ~45 kb LCRs. Published data revealed a non-pathogenic inversion polymorphism involving the NPHP1 gene flanked by two inverted ~358 kb LCRs. Using optical mapping and array-comparative genomic hybridization, we identified three potential novel structural variant (SV) haplotypes at the NPHP1 locus that may protect a haploid genome from the NPHP1 deletion. Inter-species comparative genomic analyses among primate genomes revealed massive genomic changes during evolution. The aggregated data suggest that dynamic genomic rearrangements occurred historically within the NPHP1 locus and generated SV haplotypes observed in the human population today, which may confer differential susceptibility to genomic instability and the NPHP1 deletion within a personal genome. Our study documents diverse SV haplotypes at a complex LCR-laden human genomic region. Comparative analyses provide a model for how this complex region arose during primate evolution, and studies among humans suggest that intra-species polymorphism may potentially modulate an individual’s susceptibility to acquiring disease-associated alleles. PMID:26641089

  8. Global Geometric Morphometric Analyses of the Human Pelvis Reveal Substantial Neutral Population History Effects, Even across Sexes

    PubMed Central

    Betti, Lia; von Cramon-Taubadel, Noreen; Manica, Andrea; Lycett, Stephen J.

    2013-01-01

    Recent applications of population genetic models to human craniodental traits have revealed a strong neutral component to patterns of global variation. However, little work has been undertaken to determine whether neutral processes might also be influencing the postcranium, perhaps due to substantial evidence for selection and plastic environmental responses in these regions. Recent work has provided evidence for neutral effects in the pelvis, but has been limited in regard to shape data (small numbers of linear measurements) and restricted only to males. Here, we use geometric morphometric methods to examine population variation in the human os coxae (pelvic bone) in both males and females. Neutrality is examined via apportionment of variance patterns and fit to an Out-of-Africa serial founder effect model, which is known to structure neutral genetic patterns. Moreover, we compare males and females directly, and the true versus false pelvis, in order to examine potential obstetrical effects. Our results indicate evidence for substantial neutral population history effects on pelvic shape variation. They also reveal evidence for the effect of obstetrical constraints, but these affect males and females to equivalent extents. Our results do not deny an important role for selection in regard to specific aspects of human pelvic variation, especially in terms of features associated with body size and proportions. However, our analyses demonstrate that at a global level, the shape of the os coxae reveals substantial evidence for neutral variation. Our analyses thus indicate that population variation in the human pelvis might be used to address important questions concerning population history, just as the human cranium has done. PMID:23409086

  9. Nitrogen recovery from source-separated human urine using clinoptilolite and preliminary results of its use as fertilizer.

    PubMed

    Beler-Baykal, B; Allar, A D; Bayram, S

    2011-01-01

    The use of source separated human urine as fertilizer is one of the major suggestions of the new sanitation concept ECOSAN. Urine is rich in nitrogen, phosphorus and potassium which act as plant nutrients, however its salinity is high for agricultural and landscape purposes. Moreover, characteristics change significantly throughout storage where salinity increases to higher values as the predominant form of nitrogen shifts from urea to ammonium. Transferring nitrogen in human urine onto the natural zeolite clinoptilolite and using the subsequently recovered ammonium from the exhausted clinoptilolite for agricultural/landscape purposes is suggested as an indirect route of using urine in this work. Results reporting the outcome of the proposed process together with characterization of fresh and stored urine, and preliminary work on the application of the product on the landscape plant Ficus elastica are presented. Up to 97% of the ammonium in stored urine could be transferred onto clinoptilolite through ion exchange and about 88% could be recovered subsequently from exhausted clinoptilolite, giving an overall recovery of 86%. Another important merit of the suggested process was the successful elimination of salinity. Preliminary experiments with Ficus elastica had shown that the product, i.e. clinoptilolite exhausted with ammonium, was compatible with the synthetic fertilizer tested.

  10. Molecular analyses of human induced pluripotent stem cells and embryonic stem cells

    PubMed Central

    Chin, Mark H.; Pellegrini, Matteo; Plath, Kathrin; Lowry, William E.

    2012-01-01

    Recent work from our group and others has argued that human induced pluripotent stem cells (hiPSCs) generated by the introduction of four viruses bearing reprogramming factors differ from human embryonic stem cells (hESCs) at the level of gene expression. Many of the differences seen were common across independent labs and, at least to some extent, are thought to be a result of residual expression of donor cell-specific genes (Chin et al., 2009; Ghosh et al., 2010; Marchetto et al., 2009). Two new reports re-analyze similar expression datasets as those used in Chin et al., (Chin et al., 2009) and come to different conclusions (Newman et al., 2010, Guenther et al., 2010). Here, we compare various approaches to perform gene expression meta-analysis that all support our original conclusions and present new data to demonstrate that polycistronic delivery of the reprogramming factors and extended culture brings hiPSCs transcriptionally much closer to hESCs than older methods. PMID:20682452

  11. Structural and functional analyses of human cerebral cortex using a surface-based atlas

    NASA Technical Reports Server (NTRS)

    Van Essen, D. C.; Drury, H. A.

    1997-01-01

    We have analyzed the geometry, geography, and functional organization of human cerebral cortex using surface reconstructions and cortical flat maps of the left and right hemispheres generated from a digital atlas (the Visible Man). The total surface area of the reconstructed Visible Man neocortex is 1570 cm2 (both hemispheres), approximately 70% of which is buried in sulci. By linking the Visible Man cerebrum to the Talairach stereotaxic coordinate space, the locations of activation foci reported in neuroimaging studies can be readily visualized in relation to the cortical surface. The associated spatial uncertainty was empirically shown to have a radius in three dimensions of approximately 10 mm. Application of this approach to studies of visual cortex reveals the overall patterns of activation associated with different aspects of visual function and the relationship of these patterns to topographically organized visual areas. Our analysis supports a distinction between an anterior region in ventral occipito-temporal cortex that is selectively involved in form processing and a more posterior region (in or near areas VP and V4v) involved in both form and color processing. Foci associated with motion processing are mainly concentrated in a region along the occipito-temporal junction, the ventral portion of which overlaps with foci also implicated in form processing. Comparisons between flat maps of human and macaque monkey cerebral cortex indicate significant differences as well as many similarities in the relative sizes and positions of cortical regions known or suspected to be homologous in the two species.

  12. Meta-Analyses of Human Cell-Based Cardiac Regeneration Therapies: Controversies in Meta-Analyses Results on Cardiac Cell-Based Regenerative Studies.

    PubMed

    Gyöngyösi, Mariann; Wojakowski, Wojciech; Navarese, Eliano P; Moye, Lemuel À

    2016-04-15

    In contrast to multiple publication-based meta-analyses involving clinical cardiac regeneration therapy in patients with recent myocardial infarction, a recently published meta-analysis based on individual patient data reported no effect of cell therapy on left ventricular function or clinical outcome. A comprehensive review of the data collection, statistics, and the overall principles of meta-analyses provides further clarification and explanation for this controversy. The advantages and pitfalls of different types of meta-analyses are reviewed here. Each meta-analysis approach has a place when pivotal clinical trials are lacking and sheds light on the magnitude of the treatment in a complex healthcare field.

  13. The purification, crystallization and preliminary structural characterization of human MAWDBP, a member of the phenazine biosynthesis-like protein family

    SciTech Connect

    Herde, Petra; Blankenfeldt, Wulf

    2006-06-01

    The purification, crystallization and preliminary structural characterization of human MAWD-binding protein (MAWDBP) are described. MAWDBP is the only representative of the phenazine biosynthesis-like protein family in the human genome. Its expression is elevated in several disease processes, including insulin resistance, folate deficiency and hypotension, and it may also be involved in carcinogenesis. The exact molecular function of MAWDBP is unknown. Native and seleno-l-methionine-labelled MAWDBP were expressed in Escherichia coli and crystallized at room temperature from precipitants containing 10 mM KF, 14%(w/v) PEG 3350 and 0.1 M sodium citrate pH 5.4. Crystals belong to space group H32, with unit-cell parameters a = b = 187, c = 241 Å, indicative of three to five monomers per asymmetric unit. Crystals were cryoprotected with 15%(v/v) glycerol and data have been collected to 2.7 Å resolution.

  14. Prognostic significance of CD44 in human colon cancer and gastric cancer: Evidence from bioinformatic analyses

    PubMed Central

    Xia, Pu; Xu, Xiao-Yan

    2016-01-01

    CD44 is a well-recognized stem cell biomarker expressed in colon and gastric cancer. In order to identify whether CD44 mRNA could be used as a prognostic marker in colon and gastric cancer, bioinformatic analyses were used in this study. cBioPortal analysis and COSMIC analysis were used to explore the CD44 mutation. CD44 mRNA levels were evaluated by using SAGE Genie tools and Oncomine analysis. Kaplan-Meier Plotter was performed to identify the prognostic roles of CD44 mRNA in these two cancers. In this study, first, we found that low alteration frequency of CD44 mRNA in colon and gastric cancer. Second, the high CD44 mRNA level was found in colon and gastric cancer, and it correlated with a benign survival rate in gastric cancer. Third, CD4 and CD74 may be used as markers to predict the prognosis of colon and gastric cancer. However, the deep mechanism(s) of these results remains unclear, further studies have to be performed in the future. PMID:27323782

  15. Further analyses of human kidney cell populations separated on the Space Shuttle

    NASA Technical Reports Server (NTRS)

    Stewart, Robin M.; Todd, Paul; Cole, Kenneth D.; Morrison, Dennis R.

    1992-01-01

    Cultured human embryonic kidney cells were separated into electrophoretic subpopulations in laboratory experiments and in two separation experiments on the STS-8 (Challenger) Space Shuttle flight using the mid-deck Continuous Flow Electrophoretic Separator (CFES). Populations of cells from each fraction were cultured for the lifetime of the cells, and supernatant medium was withdrawn and replaced at 4-day intervals. Withdrawn medium was frozen at -120 C for subsequent analysis. Enzyme assays, antibodies and gel electrophoresis were used as analytical tools for the detection and quantization of plasminogen activators in these samples. These assays of frozen-culture supernatant fluids confirmed the electrophoretic separation of plasminogen-activator-producing cells from nonproducing cells, the isolation of cells capable of sustained production, and the separation of cells that produce different plasminogen activators from one other.

  16. Further analyses of human kidney cell populations separated on the space shuttle

    NASA Astrophysics Data System (ADS)

    Stewart, Robin M.; Todd, Paul; Cole, Kenneth D.; Morrison, Dennis R.

    Cultured human embryonic kidney cells were separated into electrophoretic subpopulations in laboratory experiments and in two separation experiments on the STS-8 (Challenger) Space Shuttle flight using the mid-deck Continuous Flow Electrophoretic Separator (CFES). Populations of cells from each fraction were cultured for the lifetime of the cells, and supernatant medium was withdrawn and replaced at 4-day intervals. Withdrawn medium was frozen at -120°C for subsequent analysis. Enzyme assays, antibodies and gel electrophoresis were used as analytical tools for the detection and quantitation of plasminogen activators in these samples. These assays of frozen culture supernatant fluids confirmed the electrophoretic separation of plasminogen-activator producing cells from non-producing cells, the isolation of cells capable of sustained production, and the separation of cells that produce different plasminogen activators from one another.

  17. Human skin permeation of neutral species and ionic species: extended linear free-energy relationship analyses.

    PubMed

    Zhang, Keda; Chen, Ming; Scriba, Gerhard K E; Abraham, Michael H; Fahr, Alfred; Liu, Xiangli

    2012-06-01

    The permeability, K(p), of some ionized solutes (including nine acids and nine bases) through human epidermis membrane was measured in this work. Combined with the experimental K(p) data set for neutral species created by Abraham and Martins and reliable K(p) data for ionic species from the literature, a linear free-energy relationship (LFER) analysis was conducted. The values of log K(p) for 118 compounds have been correlated with solute descriptors to yield an LFER equation that incorporates neutral species and ionic species, with R(2) = 0.861 and SD = 0.462 log units. The equation can be used to predict K(p) for neutral species and ionic species, as well as partly ionized solutes. Predicted values for the passive permeation of the sodium ion and the tetraethylammonium ion are in good accord with the experimental values. It was observed that neutral acids and bases are more permeable than their ionized forms, and that the ratio depends on the actual structure. The correlation between human skin permeation and water-organic solvent/artificial membrane partitions was investigated by comparison of the coefficients in the LFER equations. Partition into cerasome is a reasonable model for partition into skin, and using cerasome as a surrogate for the partitioning process, we separate permeation into partition and diffusion processes. We show that the poor permeability of ionic species is largely due to slow diffusion through the stratum corneum. This is especially marked for a number of protonated base cations.

  18. Fluorescent in situ hybridization analyses of human oocytes in trisomy 18 and 21

    SciTech Connect

    Cheng, E.Y.; Chen, Y.J.; Gartler, S.M.

    1994-09-01

    The commonly accepted view of synapsis is that only 2 homologues can synapse at any one site and that this restriction applies to polyploids as well. However, triple synapsis has been observed is some triploid plants and in triploid chicken. In humans, triple synapsis of the long arm of chromosome 21 was detected in sperm of a trisomic 21 individual. More recently, studies of oocytes from trisomic 21 and 18 fetuses also indicated extensive triple synapsis along the entire length of the chromosomes. To further investigate this question, we undertook an evaluation of trivalent synapsis in fetal oocytes from 2 trisomic 21 and 2 trisomic 18 fetuses using fluorescent in situ hybridization (FISH) with whole chromosome probes. Oocytes were hybridized with whole chromosome probes obtained from ONCOR, Inc. after fixation with methanol and acetic acid. Slides were scored for the distribution of prophase stages, hybridization efficiency, and hybridization characteristics of chromosomes 18 and 21 in the trisomic 18 and 21 fetuses respectively. Fifty-eight per cent (379/650) of pachytenes analyzed for chromosome 18 contained a conspicous trivalent and 319 (48%) of these nuclei contained a single, thick, continuous fluorescent signal consistent with complete triple synapsis along the entire length of all 3 chromosomes. Sixteen per cent (104/650) of pachytene contained 2 signals consistent with a bivalent and a univalent, and 9 cells contained 3 thin signals consistent with asynapsis of all 3 chromosomes. The remaining 158 pachytenes had unusual pairing configurations that we could not classify, but they most likely represent trivalents with partial pairing between different homologues. In the 2 trisomic 21 fetuses, the majority (143/232) of pachytenes also contained one signal while only 52 cells contained a bivalent and univalent. Five cells contained 3 separate signals. These results confirm the existence of triple synapsis in human meiosis.

  19. Analysing inter-relationships among water, governance, human development variables in developing countries

    NASA Astrophysics Data System (ADS)

    Dondeynaz, C.; Carmona Moreno, C.; Céspedes Lorente, J. J.

    2012-10-01

    The "Integrated Water Resources Management" principle was formally laid down at the International Conference on Water and Sustainable development in Dublin 1992. One of the main results of this conference is that improving Water and Sanitation Services (WSS), being a complex and interdisciplinary issue, passes through collaboration and coordination of different sectors (environment, health, economic activities, governance, and international cooperation). These sectors influence or are influenced by the access to WSS. The understanding of these interrelations appears as crucial for decision makers in the water sector. In this framework, the Joint Research Centre (JRC) of the European Commission (EC) has developed a new database (WatSan4Dev database) containing 42 indicators (called variables in this paper) from environmental, socio-economic, governance and financial aid flows data in developing countries. This paper describes the development of the WatSan4Dev dataset, the statistical processes needed to improve the data quality, and finally, the analysis to verify the database coherence is presented. Based on 25 relevant variables, the relationships between variables are described and organised into five factors (HDP - Human Development against Poverty, AP - Human Activity Pressure on water resources, WR - Water Resources, ODA - Official Development Aid, CEC - Country Environmental Concern). Linear regression methods are used to identify key variables having influence on water supply and sanitation. First analysis indicates that the informal urbanisation development is an important factor negatively influencing the percentage of the population having access to WSS. Health, and in particular children's health, benefits from the improvement of WSS. Irrigation is also enhancing Water Supply service thanks to multi-purpose infrastructure. Five country profiles are also created to deeper understand and synthetize the amount of information gathered. This new

  20. Comparative genomic analyses of the human fungal pathogens Coccidioides and their relatives

    PubMed Central

    Sharpton, Thomas J.; Stajich, Jason E.; Rounsley, Steven D.; Gardner, Malcolm J.; Wortman, Jennifer R.; Jordar, Vinita S.; Maiti, Rama; Kodira, Chinnappa D.; Neafsey, Daniel E.; Zeng, Qiandong; Hung, Chiung-Yu; McMahan, Cody; Muszewska, Anna; Grynberg, Marcin; Mandel, M. Alejandra; Kellner, Ellen M.; Barker, Bridget M.; Galgiani, John N.; Orbach, Marc J.; Kirkland, Theo N.; Cole, Garry T.; Henn, Matthew R.; Birren, Bruce W.; Taylor, John W.

    2009-01-01

    While most Ascomycetes tend to associate principally with plants, the dimorphic fungi Coccidioides immitis and Coccidioides posadasii are primary pathogens of immunocompetent mammals, including humans. Infection results from environmental exposure to Coccidiodies, which is believed to grow as a soil saprophyte in arid deserts. To investigate hypotheses about the life history and evolution of Coccidioides, the genomes of several Onygenales, including C. immitis and C. posadasii; a close, nonpathogenic relative, Uncinocarpus reesii; and a more diverged pathogenic fungus, Histoplasma capsulatum, were sequenced and compared with those of 13 more distantly related Ascomycetes. This analysis identified increases and decreases in gene family size associated with a host/substrate shift from plants to animals in the Onygenales. In addition, comparison among Onygenales genomes revealed evolutionary changes in Coccidioides that may underlie its infectious phenotype, the identification of which may facilitate improved treatment and prevention of coccidioidomycosis. Overall, the results suggest that Coccidioides species are not soil saprophytes, but that they have evolved to remain associated with their dead animal hosts in soil, and that Coccidioides metabolism genes, membrane-related proteins, and putatively antigenic compounds have evolved in response to interaction with an animal host. PMID:19717792

  1. Linear time delay methods and stability analyses of the human spine. Effects of neuromuscular reflex response.

    PubMed

    Franklin, Timothy C; Granata, Kevin P; Madigan, Michael L; Hendricks, Scott L

    2008-08-01

    Linear stability methods were applied to a biomechanical model of the human musculoskeletal spine to investigate effects of reflex gain and reflex delay on stability. Equations of motion represented a dynamic 18 degrees-of-freedom rigid-body model with time-delayed reflexes. Optimal muscle activation levels were identified by minimizing metabolic power with the constraints of equilibrium and stability with zero reflex time delay. Muscle activation levels and associated muscle forces were used to find the delay margin, i.e., the maximum reflex delay for which the system was stable. Results demonstrated that stiffness due to antagonistic co-contraction necessary for stability declined with increased proportional reflex gain. Reflex delay limited the maximum acceptable proportional reflex gain, i.e., long reflex delay required smaller maximum reflex gain to avoid instability. As differential reflex gain increased, there was a small increase in acceptable reflex delay. However, differential reflex gain with values near intrinsic damping caused the delay margin to approach zero. Forward-dynamic simulations of the fully nonlinear time-delayed system verified the linear results. The linear methods accurately found the delay margin below which the nonlinear system was asymptotically stable. These methods may aid future investigations in the role of reflexes in musculoskeletal stability.

  2. Analyses of the secondary particle radiation and the DNA damage it causes to human keratinocytes

    SciTech Connect

    Lebel E.; Rusek A.; Sivertz, M.; Yip, K.; Thompson, K.; Tafrov, S.

    2011-11-22

    High-energy protons, and high mass and energy ions, along with the secondary particles they produce, are the main contributors to the radiation hazard during space explorations. Skin, particularly the epidermis, consisting mainly of keratinocytes with potential for proliferation and malignant transformation, absorbs the majority of the radiation dose. Therefore, we used normal human keratinocytes to investigate and quantify the DNA damage caused by secondary radiation. Its manifestation depends on the presence of retinol in the serum-free media, and is regulated by phosphatidylinositol 3-kinases. We simulated the generation of secondary radiation after the impact of protons and iron ions on an aluminum shield. We also measured the intensity and the type of the resulting secondary particles at two sample locations; our findings agreed well with our predictions. We showed that secondary particles inflict DNA damage to different extents, depending on the type of primary radiation. Low-energy protons produce fewer secondary particles and cause less DNA damage than do high-energy protons. However, both generate fewer secondary particles and inflict less DNA damage than do high mass and energy ions. The majority of cells repaired the initial damage, as denoted by the presence of 53BPI foci, within the first 24 hours after exposure, but some cells maintained the 53BP1 foci longer.

  3. Analyses of the Secondary Particle Radiation and the DNA Damage it Causes to Human Keratinocytes

    SciTech Connect

    Lebel E. A.; Tafrov S.; Rusek, A.; Sivertz, M. B.; Yip, K.; Thompson, K. H.

    2011-11-01

    High-energy protons, and high mass and energy ions, along with the secondary particles they produce, are the main contributors to the radiation hazard during space explorations. Skin, particularly the epidermis, consisting mainly of keratinocytes with potential for proliferation and malignant transformation, absorbs the majority of the radiation dose. Therefore, we used normal human keratinocytes to investigate and quantify the DNA damage caused by secondary radiation. Its manifestation depends on the presence of retinol in the serum-free media, and is regulated by phosphatidylinositol 3-kinases. We simulated the generation of secondary radiation after the impact of protons and iron ions on an aluminum shield. We also measured the intensity and the type of the resulting secondary particles at two sample locations; our findings agreed well with our predictions. We showed that secondary particles inflict DNA damage to different extents, depending on the type of primary radiation. Low-energy protons produce fewer secondary particles and cause less DNA damage than do high-energy protons. However, both generate fewer secondary particles and inflict less DNA damage than do high mass and energy ions. The majority of cells repaired the initial damage, as denoted by the presence of 53BPI foci, within the first 24 hours after exposure, but some cells maintained the 53BP1 foci longer.

  4. Gene Sequence Analyses of the Healthy Oral Microbiome in Humans and Companion Animals.

    PubMed

    Davis, Eric M

    2016-06-01

    It has long been accepted that certain oral bacterial species are responsible for the development of periodontal disease. However, the focus of microbial and immunological research is shifting from studying the organisms associated with disease to examining the indigenous microbial inhabitants that are present in health. Microbiome refers to the aggregate genetic material of all microorganisms living in, or on, a defined habitat. Recent developments in gene sequence analysis have enabled detection and identification of bacteria from polymicrobial samples, including subgingival plaque. Diversity surveys utilizing this technology have demonstrated that bacterial culture techniques have vastly underestimated the richness and diversity of microorganisms in vivo, since only certain bacteria grow in vitro. Surveys using gene sequence analysis have demonstrated that the healthy oral microbiome is composed of an unexpectedly high number of diverse species, including putative pathogens. These findings support the view that coevolution microorganisms and macroscopic hosts has occurred in which certain microorganisms have adapted to survive in the oral cavity and host immune tolerance has allowed the establishment of a symbiotic relationship in which both parties receive benefits (mutualism). This review describes gene sequence analysis as an increasingly common, culture-independent tool for detecting bacteria in vivo and describes the results of recent oral microbiome diversity surveys of clinically healthy humans, dogs, and cats. Six bacterial phyla consistently dominated the healthy oral microbiome of all 3 host species. Previous hypotheses on etiology of periodontitis are reviewed in light of new scientific findings. Finally, the consideration that clinically relevant periodontal disease occurs when immune tolerance of the symbiotic oral microbiome is altered to a proinflammatory response will be discussed.

  5. Comparative Genetic Analyses of Human Rhinovirus C (HRV-C) Complete Genome from Malaysia.

    PubMed

    Khaw, Yam Sim; Chan, Yoke Fun; Jafar, Faizatul Lela; Othman, Norlijah; Chee, Hui Yee

    2016-01-01

    Human rhinovirus-C (HRV-C) has been implicated in more severe illnesses than HRV-A and HRV-B, however, the limited number of HRV-C complete genomes (complete 5' and 3' non-coding region and open reading frame sequences) has hindered the in-depth genetic study of this virus. This study aimed to sequence seven complete HRV-C genomes from Malaysia and compare their genetic characteristics with the 18 published HRV-Cs. Seven Malaysian HRV-C complete genomes were obtained with newly redesigned primers. The seven genomes were classified as HRV-C6, C12, C22, C23, C26, C42, and pat16 based on the VP4/VP2 and VP1 pairwise distance threshold classification. Five of the seven Malaysian isolates, namely, 3430-MY-10/C22, 8713-MY-10/C23, 8097-MY-11/C26, 1570-MY-10/C42, and 7383-MY-10/pat16 are the first newly sequenced complete HRV-C genomes. All seven Malaysian isolates genomes displayed nucleotide similarity of 63-81% among themselves and 63-96% with other HRV-Cs. Malaysian HRV-Cs had similar putative immunogenic sites, putative receptor utilization and potential antiviral sites as other HRV-Cs. The genomic features of Malaysian isolates were similar to those of other HRV-Cs. Negative selections were frequently detected in HRV-Cs complete coding sequences indicating that these sequences were under functional constraint. The present study showed that HRV-Cs from Malaysia have diverse genetic sequences but share conserved genomic features with other HRV-Cs. This genetic information could provide further aid in the understanding of HRV-C infection.

  6. Comparative Genetic Analyses of Human Rhinovirus C (HRV-C) Complete Genome from Malaysia

    PubMed Central

    Khaw, Yam Sim; Chan, Yoke Fun; Jafar, Faizatul Lela; Othman, Norlijah; Chee, Hui Yee

    2016-01-01

    Human rhinovirus-C (HRV-C) has been implicated in more severe illnesses than HRV-A and HRV-B, however, the limited number of HRV-C complete genomes (complete 5′ and 3′ non-coding region and open reading frame sequences) has hindered the in-depth genetic study of this virus. This study aimed to sequence seven complete HRV-C genomes from Malaysia and compare their genetic characteristics with the 18 published HRV-Cs. Seven Malaysian HRV-C complete genomes were obtained with newly redesigned primers. The seven genomes were classified as HRV-C6, C12, C22, C23, C26, C42, and pat16 based on the VP4/VP2 and VP1 pairwise distance threshold classification. Five of the seven Malaysian isolates, namely, 3430-MY-10/C22, 8713-MY-10/C23, 8097-MY-11/C26, 1570-MY-10/C42, and 7383-MY-10/pat16 are the first newly sequenced complete HRV-C genomes. All seven Malaysian isolates genomes displayed nucleotide similarity of 63–81% among themselves and 63–96% with other HRV-Cs. Malaysian HRV-Cs had similar putative immunogenic sites, putative receptor utilization and potential antiviral sites as other HRV-Cs. The genomic features of Malaysian isolates were similar to those of other HRV-Cs. Negative selections were frequently detected in HRV-Cs complete coding sequences indicating that these sequences were under functional constraint. The present study showed that HRV-Cs from Malaysia have diverse genetic sequences but share conserved genomic features with other HRV-Cs. This genetic information could provide further aid in the understanding of HRV-C infection. PMID:27199901

  7. Morphometric and colorimetric analyses of human tumor cell line growth and drug sensitivity in soft agar culture.

    PubMed

    Alley, M C; Pacula-Cox, C M; Hursey, M L; Rubinstein, L R; Boyd, M R

    1991-02-15

    Previous studies have demonstrated the suitability of image analysis of tetrazolium-stained colonies to assess growth and drug sensitivity of human tumor cells cultivated in soft agar culture. In the present study, the potential utility of colorimetric analysis to expedite experimental drug evaluations using human tumor cell lines was investigated. The same culture dishes were assessed by image analysis and by formazan colorimetry for purposes of comparing multiple methods of measuring growth as well as growth inhibition. Replicate cultures treated with 2-(p-iodonitrophenyl)-3-p-nitrophenyl-5-phenyltetrazolium chloride or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide exhibited nearly identical colony count and volume indices as well as excellent correlation in colorimetric end points. Colony-forming unit volume analysis versus colorimetric assessment of the same cultures following dimethyl sulfoxide extraction of protamine sulfate-rinsed, dried soft agar cultures exhibited excellent linear correlation for both growth (Pearson r ranging from 0.95 to 1.00) and drug sensitivity (Pearson r ranging from 0.90 to 0.99, and Spearman r ranging from 0.82 to 0.97) and similar drug sensitivity profiles. Results of the current investigation indicate that end points of soft agar culture remain stable for a period of at least 2 weeks following assay termination. In addition, a colorimetric detection range of 1.3-2.2 log units permits determinations of survival levels ranging from 100 to 5% of respective control levels. Colorimetric analysis is anticipated to expedite soft agar colony formation assay evaluations (a) by reducing the need to use the more rigorous and time-consuming image analysis procedures to measure activity in preliminary drug sensitivity assays and (b) by permitting the determination of effective concentration ranges of new experimental agents for subsequent, more detailed investigations.

  8. Genetic analyses of the human eye colours using a novel objective method for eye colour classification.

    PubMed

    Andersen, Jeppe D; Johansen, Peter; Harder, Stine; Christoffersen, Susanne R; Delgado, Mikaela C; Henriksen, Sarah T; Nielsen, Mette M; Sørensen, Erik; Ullum, Henrik; Hansen, Thomas; Dahl, Anders L; Paulsen, Rasmus R; Børsting, Claus; Morling, Niels

    2013-09-01

    In this study, we present a new objective method for measuring the eye colour on a continuous scale that allows researchers to associate genetic markers with different shades of eye colour. With the use of the custom designed software Digital Iris Analysis Tool (DIAT), the iris was automatically identified and extracted from high resolution digital images. DIAT was made user friendly with a graphical user interface. The software counted the number of blue and brown pixels in the iris image and calculated a Pixel Index of the Eye (PIE-score) that described the eye colour quantitatively. The PIE-score ranged from -1 to 1 (brown to blue). The software eliminated the need for user based interpretation and qualitative eye colour categories. In 94% (570) of 605 analyzed eye images, the iris region was successfully extracted and a PIE-score was calculated. A very high correlation between the PIE-score and the human perception of eye colour was observed. The correlations between the PIE-scores and the six IrisPlex SNPs (HERC2 rs12913832, OCA2 rs1800407, SLC24A4 rs12896399, TYR rs1393350, SLC45A2 rs16891982 and IRF4 rs12203592) were analyzed in 570 individuals. Significant differences (p<10(-6)) in the PIE-scores of the individuals typed as HERC2 rs12913832 G (PIE=0.99) and rs12913832 GA (PIE=-0.71) or A (PIE=-0.87) were observed. We adjusted for the effect of HERC2 rs12913832 and showed that the quantitative PIE-scores were significantly associated with SNPs with minor effects (OCA2 rs1800407, SLC24A4 rs12896399 and TYR rs1393350) on the eye colour. We evaluated the two published prediction models for eye colour (IrisPlex [1] and Snipper[2]) and compared the predictions with the PIE-scores. We found good concordance with the prediction from individuals typed as HERC2 rs12913832 G. However, both methods had difficulties in categorizing individuals typed as HERC2 rs12913832 GA because of the large variation in eye colour in HERC2 rs12913832 GA individuals. With the use of

  9. The Neogene Ogallala Formation in Southwestern Kansas and Northeastern New Mexico: Preliminary Magnetostratigraphic Analyses for the High Plains-Ogallala Drilling Program

    NASA Astrophysics Data System (ADS)

    Zeigler, K. E.; Petronis, M. S.; Smith, J. J.; Ludvigson, G. A.; Doveton, J.

    2012-12-01

    A better understanding of the Ogallala Formation is critical in terms of refining groundwater flow models and management policies for communities relying on aquifers in heterolithic sequences around the globe. The High Plains aquifer remains under increasing stress with the growth of both urban and agricultural areas and therefore, developing the best groundwater management policies will depend on the most accurate characterization of the aquifer, the aquifer materials and their stratigraphic and geochronologic framework. Although mammalian faunal assemblages and volcanic ash bed tephrochronology provide a basic geochronologic framework for the Ogallala Formation, better precision in terms of correlation is needed in order to understand formative processes and depositional histories for the primary water-bearing units in the High Plains aquifer and confining strata. The High Plains-Ogallala Drilling Program (HPODP) was developed to advance understanding of sedimentary facies, stratigraphic framework, and chronostratigraphy of the Ogallala Formation and overlying units that comprise the central High Plains aquifer. The drilling program began in the early summer of 2011 in Haskell Co., western Kansas. By early fall 2011; the drill crew was at 92 m with about 32 m to go until they expect to hit bedrock. The final 32 meters of core was extracted during the summer 2012. Here we report the preliminary magnetostratigraphic and rock magnetic data from the first section of core. We scanned the entire 92 meters of core using an ASC Core Analysis System with a Bartington Instruments MS2C magnetic susceptibility coil allowing for bulk susceptibility measurements to be obtained along the length of the core. In addition, we collected 40 sub samples for paleomagnetic and rock magnetic. Bulk susceptibility data reveal depth dependent changes in rock magnetic properties that we interpret to reflect either climatic driven variations impacting the depositional system or a change in

  10. Genetic Structure and Preliminary Findings of Cryptic Diversity of the Malaysian Mahseer (Tor tambroides Valenciennes: Cyprinidae) Inferred from Mitochondrial DNA and Microsatellite Analyses

    PubMed Central

    Abdul Rahim, Khairul Adha

    2013-01-01

    This study examines the population genetic structure of Tor tambroides, an important freshwater fish species in Malaysia, using fifteen polymorphic microsatellite loci and sequencing of 464 base pairs of the mitochondrial cytochrome c oxidase I (COI) gene. A total of 152 mahseer samples were collected from eight populations throughout the Malaysia river system. Microsatellites results found high levels of intrapopulation variations, but mitochondrial COI results found high levels of interpopulations differentiation. The possible reasons for their discrepancies might be the varying influence of genetic drift on each marker or the small sample sizes used in most of the populations. The Kelantan population showed very low levels of genetic variations using both mitochondrial and microsatellite analyses. Phylogenetic analysis of the COI gene found a unique haplotype (ER8∗), possibly representing a cryptic lineage of T. douronensis, from the Endau-Rompin population. Nevertheless, the inclusion of nuclear microsatellite analyses could not fully resolve the genetic identity of haplotype ER8∗ in the present study. Overall, the findings showed a serious need for more comprehensive and larger scale samplings, especially in remote river systems, in combination with molecular analyses using multiple markers, in order to discover more cryptic lineages or undescribed “genetic species” of mahseer. PMID:24455674

  11. Direct Dating and Physico-Chemical Analyses Cast Doubts on the Coexistence of Humans and Dwarf Hippos in Cyprus

    PubMed Central

    Zazzo, Antoine; Lebon, Matthieu; Quiles, Anita; Reiche, Ina; Vigne, Jean-Denis

    2015-01-01

    In the Mediterranean, the island dwarf megafaunas became extinct around the end of the Pleistocene, during a period of rapid and global climate change. In Cyprus, this coincided with the first human presence on the island, as attested by the rock shelter of Akrotiri-Aetokremnos where an Epipaleolithic anthropogenic layer (stratum 2) was found overlying a massive accumulation of pygmy hippopotamus (Phanourios minor (Desmarest, 1822)) [Boekschoten and Sondaar, 1972] bones (stratum 4). The relationship between the two layers is highly controversial and the role played by humans in hippo extinction remains fiercely debated. Here, we provide new, direct radiocarbon and physico-chemical analyses on calcined bones which elucidates the complex depositional history of the assemblage. Bone turquoise was identified using micro-PIXE analysis and depth-profiling together with Vis spectroscopy, demonstrating that these bones were not freshly burned. Bayesian modeling of the radiocarbon dates indicates that stratum 4 accumulated during the first half of the 13th mill cal BP and that calcination occurred several hundred years later. We conclude that accumulation occurred naturally during the beginning of the Younger Dryas and that Epipalaeolithic visitors subsequently used the bones as fuel, starting from the mid-13th mill cal BP. At that time, dwarf hippos were probably already extinct or at least highly endangered. Our results shed new light on the possible causes of hippo extinction, on the subsequent introduction of the wild boar and on the earliest occupation of the island by humans. PMID:26284623

  12. Direct Dating and Physico-Chemical Analyses Cast Doubts on the Coexistence of Humans and Dwarf Hippos in Cyprus.

    PubMed

    Zazzo, Antoine; Lebon, Matthieu; Quiles, Anita; Reiche, Ina; Vigne, Jean-Denis

    2015-01-01

    In the Mediterranean, the island dwarf megafaunas became extinct around the end of the Pleistocene, during a period of rapid and global climate change. In Cyprus, this coincided with the first human presence on the island, as attested by the rock shelter of Akrotiri-Aetokremnos where an Epipaleolithic anthropogenic layer (stratum 2) was found overlying a massive accumulation of pygmy hippopotamus (Phanourios minor (Desmarest, 1822)) [Boekschoten and Sondaar, 1972] bones (stratum 4). The relationship between the two layers is highly controversial and the role played by humans in hippo extinction remains fiercely debated. Here, we provide new, direct radiocarbon and physico-chemical analyses on calcined bones which elucidates the complex depositional history of the assemblage. Bone turquoise was identified using micro-PIXE analysis and depth-profiling together with Vis spectroscopy, demonstrating that these bones were not freshly burned. Bayesian modeling of the radiocarbon dates indicates that stratum 4 accumulated during the first half of the 13th mill cal BP and that calcination occurred several hundred years later. We conclude that accumulation occurred naturally during the beginning of the Younger Dryas and that Epipalaeolithic visitors subsequently used the bones as fuel, starting from the mid-13th mill cal BP. At that time, dwarf hippos were probably already extinct or at least highly endangered. Our results shed new light on the possible causes of hippo extinction, on the subsequent introduction of the wild boar and on the earliest occupation of the island by humans.

  13. Purification, crystallization and preliminary X-ray diffraction studies on human Ca2+-binding protein S100B.

    PubMed

    Ostendorp, Thorsten; Heizmann, Claus W; Kroneck, Peter M H; Fritz, Günter

    2005-07-01

    S100B, a Ca2+-binding protein, acts intracellularly as a Ca2+-signalling protein but is also secreted to the extracellular space, acting in a cytokine-like manner through its receptor RAGE. Recombinant human S100B has been purified and crystallized in the Ca2+-bound state. Size-exclusion chromatography indicates that S100B can exist as a dimer and as a multimer in solution. Crystals of S100B diffract to 1.9 A and belong to space group P2(1), with unit-cell parameters a = 63.4, b = 81.6, c = 71.5 A, alpha = 90, beta = 107, gamma = 90 degrees. Preliminary analysis of the X-ray data indicate that there are four homodimers per asymmetric unit.

  14. Purification, crystallization and preliminary X-ray analysis of the IgV domain of human nectin-4

    PubMed Central

    Xu, Xiang; Zhang, Xiaoai; Lu, Guangwen; Cai, Yongping

    2012-01-01

    Nectin-4 belongs to a family of immunoglobulin-like cell adhesion molecules and is highly expressed in cancer cells. Recently, nectin-4 was found to be a receptor of measles virus and the IgV domain sustains strong binding to measles virus H protein. In this study, the successful expression and purification of human nectin-4 V domain (nectin-4v) is reported. The purified protein was crystallized using the sitting-drop vapour-diffusion method. The crystals diffracted to 1.8 Å resolution and belonged to space group P21, with unit-cell parameters a = 33.1, b = 51.7, c = 56.9 Å, β = 94.7°. Preliminary analysis of the diffraction data was also performed. PMID:22869128

  15. Crystallization, preliminary X-ray analysis and Patterson search of a new aspartic protease isolated from human urine.

    PubMed

    Canduri, F; Teodoro, L G; Lorenzi, C C; Gomes, R A; Fontes, M R; Arni, R K; de Azevedo Júnior, W F

    1998-10-01

    Aspartic protease (EC 3.4.23) make up a widely distributed class of enzymes in animals, plants, microbes and, viruses. In animals these enzymes perform diverse functions, which range from digestion of food proteins to very specific regulatory roles. In contrast the information about the well-characterized aspartic proteases, very little is known about the corresponding enzyme in urine. A new aspartic protease isolated from human urine has been crystallized and X-ray diffraction data collected to 2.45 A resolution using a synchrotron radiation source. Crystals belong to the space group P2(1)2(1)2(1). The cell parameters obtained were a = 50.99, b = 75.56 and c = 89.90 A. Preliminary analysis revealed the presence of one molecule in the asymmetric unit. The structure was determined using the molecular replacement technique and is currently being refined using simulated annealing and conjugate gradient protocols.

  16. Control of oral human papillomavirus (HPV) by medicinal mushrooms, Trametes versicolor and Ganoderma lucidum: a preliminary clinical trial.

    PubMed

    Donatini, Bruno

    2014-01-01

    This preliminary randomized study investigated the efficacy of medicinal mushrooms, Trametes versicolor (TV), Ganoderma lucidum (GL), and Laetiporus sulphureus (LS), on the clearance of oral human papillomavirus (HPV, serotypes 16 and 18). Among 472 patients who underwent oral swabs for gingivitis, 61 patients were positive for HPV16 or HPV18. Twenty patients were included in group 1 (LS) and 41 patients were included in group 2 (TV+GL) for 2 months. Polymerase chain reaction (PCR) for HPV was performed at inclusion and after 2 months. In group 1, the clearance was equal to 5% after 2 months of treatment. In group 2, the clearance was equal to 88% (P<0.001). The detection of HPV16 or HPV18 could become relevant in routine since positivity is frequent and because a harmless and costless treatment may exist. The use of TV+GL for the clearance of oral HPV deserves further investigation.

  17. Estrogen binding and estrogen receptor activity in the human prostate: a preliminary report.

    PubMed

    Fondo, E Y; Menendez-Botet, C J; Schwartz, M K; Whitmore, W F

    1981-03-01

    Assay of estrogen receptor activity in prostates from patients who ranged in age from 22 to 78 years and had not received any previous hormonal therapy was carried out by incubation of cytosols with (3)H-estradiol in the presence and absence of excess, nonradioactive estradiol. Hyperplastic prostatic tissues were used in the study. The kinetics of each reaction were studied and analysis of the data revealed 3.4 to 35.7 femtomoles of receptor protein per mg of cytosol protein; the dissociation constants obtained from a Scatchard plot ranged from 1.1 × 10(-10) to 1.2 × 10(-8)M.The small number of patients prevents realistic quantitative assessment of the apparent estrogen binding activity demonstrated in these preliminary studies, but the qualitative identification of such activity provides possible grounds for further insight into the hormonal mechanisms in the pathophysiology of prostatic diseases and of their responses to endocrine therapy.

  18. THE NATIONAL HUMAN EXPOSURE ASSESSMENT SURVEY (NHEXAS) STUDY IN ARIZONA-INTRODUCTION AND PRELIMINARY RESULTS

    EPA Science Inventory

    The objective of the National Human Exposure Assessment Survey (NHEXAS) in Arizona is to determine the multimedia distribution of total human exposure to environmental pollutants in the classes of metals, pesticides, and volatile organic compounds (VOCs) for the population of Ari...

  19. A Preliminary Study of Peer-to-Peer Human-Robot Interaction

    NASA Technical Reports Server (NTRS)

    Fong, Terrence; Flueckiger, Lorenzo; Kunz, Clayton; Lees, David; Schreiner, John; Siegel, Michael; Hiatt, Laura M.; Nourbakhsh, Illah; Simmons, Reid; Ambrose, Robert

    2006-01-01

    The Peer-to-Peer Human-Robot Interaction (P2P-HRI) project is developing techniques to improve task coordination and collaboration between human and robot partners. Our work is motivated by the need to develop effective human-robot teams for space mission operations. A central element of our approach is creating dialogue and interaction tools that enable humans and robots to flexibly support one another. In order to understand how this approach can influence task performance, we recently conducted a series of tests simulating a lunar construction task with a human-robot team. In this paper, we describe the tests performed, discuss our initial results, and analyze the effect of intervention on task performance.

  20. Combination Of Static And Dynami,C Stereophotogrammetry For The Kinetic Analysis Of Human Locomotion: Preliminary Results

    NASA Astrophysics Data System (ADS)

    Schaer, Alex R.; Sheffer, Daniel B.; Jones, D.; Meier, G.; Baumann, Juerg U.

    1989-04-01

    For a deeper understanding of the complexity of human walking movement not only a kinematic analysis , but also a comprehensive three-dimensional biomechanical model of the human body is required to detail the kinetic activities. This research combined static stereophotogrammetric determination of body segment mass parameters with three-dimensional gait analysis by cinephotography, direct linear transformation and two force plates. A method of combining the two independent analyses by defining the anatomical axes of each segment is shown. Practical problems arising in dynamic and stereometric analysis are demonstrated. Power spectra of a normal and a matched subject with spastic diplegia were calculated for a proper design of the kinematic analysis.

  1. Is a Swine Model of Arteriovenous Malformation Suitable for Human Extracranial Arteriovenous Malformation? A Preliminary Study

    SciTech Connect

    Lv, Ming-ming; Fan, Xin-dong; Su, Li-xin

    2013-10-15

    Objective: A chronic arteriovenous malformation (AVM) model using the swine retia mirabilia (RMB) was developed and compared with the human extracranial AVM (EAVM) both in hemodynamics and pathology, to see if this brain AVM model can be used as an EAVM model. Methods: We created an arteriovenous fistula between the common carotid artery and the external jugular vein in eight animals by using end-to-end anastomosis. All animals were sacrificed 1 month after surgery, and the bilateral retia were obtained at autopsy and performed hematoxylin and eosin staining and immunohistochemistry. Pre- and postsurgical hemodynamic evaluations also were conducted. Then, the blood flow and histological changes of the animal model were compared with human EAVM. Results: The angiography after operation showed that the blood flow, like human EAVM, flowed from the feeding artery, via the nidus, drained to the draining vein. Microscopic examination showed dilated lumina and disrupted internal elastic lamina in both RMB of model and nidus of human EAVM, but the thickness of vessel wall had significant difference. Immunohistochemical reactivity for smooth muscle actin, angiopoietin 1, and angiopoietin 2 were similar in chronic model nidus microvessels and human EAVM, whereas vascular endothelial growth factor was significant difference between human EAVM and RMB of model. Conclusions: The AVM model described here is similar to human EAVM in hemodynamics and immunohistochemical features, but there are still some differences in anatomy and pathogenetic mechanism. Further study is needed to evaluate the applicability and efficacy of this model.

  2. Human-Centered Technologies and Procedures for Future Air Traffic Management: A Preliminary Overview of 1996 Studies and Results

    NASA Technical Reports Server (NTRS)

    Smith, Philip; McCoy, Elaine; Denning, Rebecca; Woods, David; Sarter, Nadine; Dekker, Sidney; Billings, Charles

    1996-01-01

    In this project, we have been exploring the use of a general methodology to predict the impact of future Air Traffic Management (ATM) concepts and technologies. In applying this methodology, our emphasis has been on the importance of modeling coordination and cooperation among the multiple agents within this system, and on understanding how the interactions among these agents will be influenced as new roles, responsibilities, procedures and technologies are introduced. To accomplish this, we have been collecting data on performance under the current air traffic management system, trying to identify critical problem areas and looking for exemplars suggestive of general approaches for solving such problems. Based on the results of these field studies, we have developed a set of scenarios centered around potential future system designs, and have conducted studies using these scenarios involving a total 40 controllers, dispatchers, pilots and traffic managers. The purpose of this report is to provide NASA with an early summary of the major recommendations that have resulted from our research under the AATT Program thus far. Recommendations 1-3 deal with general approaches that our findings suggest should be incorporated in future AATT Program activities, while Recommendations 4-11 identify some specific topics and technologies that merit research and development activities. Detailed technical reports containing supporting data, as well as the results of our still ongoing analyses, will be provided at a later date. The remainder of this report is organized as follows. Section 1 briefly describes the general design philosophy supported by our empirical studies. Section 2 presents the research methods we have used for identifying requirements for future system designs and for evaluating alternative design solutions. Section 3 discusses preliminary results from an initial set of investigations that we have conducted using these research methods. Section 4 then provides an

  3. Preliminary performance assessment for the Waste Isolation Pilot Plant, December 1992. Volume 4: Uncertainty and sensitivity analyses for 40 CFR 191, Subpart B

    SciTech Connect

    Not Available

    1993-08-01

    Before disposing of transuranic radioactive waste in the Waste Isolation Pilot Plant (WIPP), the United States Department of Energy (DOE) must evaluate compliance with applicable long-term regulations of the United States Environmental Protection Agency (EPA). Sandia National Laboratories is conducting iterative performance assessments (PAs) of the WIPP for the DOE to provide interim guidance while preparing for a final compliance evaluation. This volume of the 1992 PA contains results of uncertainty and sensitivity analyses with respect to the EPA`s Environmental Protection Standards for Management and Disposal of Spent Nuclear Fuel, High-Level and Transuranic Radioactive Wastes (40 CFR 191, Subpart B). Additional information about the 1992 PA is provided in other volumes. Results of the 1992 uncertainty and sensitivity analyses indicate that, conditional on the modeling assumptions, the choice of parameters selected for sampling, and the assigned parameter-value distributions, the most important parameters for which uncertainty has the potential to affect compliance with 40 CFR 191B are: drilling intensity, intrusion borehole permeability, halite and anhydrite permeabilities, radionuclide solubilities and distribution coefficients, fracture spacing in the Culebra Dolomite Member of the Rustler Formation, porosity of the Culebra, and spatial variability of Culebra transmissivity. Performance with respect to 40 CFR 191B is insensitive to uncertainty in other parameters; however, additional data are needed to confirm that reality lies within the assigned distributions.

  4. Crystallization and preliminary X-ray diffraction analysis of human phosphate-binding protein

    SciTech Connect

    Contreras-Martel, Carlos; Carpentier, Philippe; Morales, Renaud; Renault, Frédérique; Chesne-Seck, Marie-Laure; Rochu, Daniel; Masson, Patrick; Fontecilla-Camps, Juan Carlos; Chabrière, Eric

    2006-01-01

    The purification, detergent-exchange protocol and crystallization conditions that led to the discovery of HPBP are reported. HPBP is a new human apoprotein that is absent from the genomic database and is the first phosphate transporter characterized in human plasma. Human phosphate-binding protein (HPBP) was serendipitously discovered by crystallization and X-ray crystallography. HPBP belongs to a eukaryotic protein family named DING that is systematically absent from the genomic database. This apoprotein of 38 kDa copurifies with the HDL-associated apoprotein paraoxonase (PON1) and binds inorganic phosphate. HPBP is the first identified transporter capable of binding phosphate ions in human plasma. Thus, it may be regarded as a predictor of phosphate-related diseases such as atherosclerosis. In addition, HPBP may be a potential therapeutic protein for the treatment of such diseases. Here, the purification, detergent-exchange protocol and crystallization conditions that led to the discovery of HPBP are reported.

  5. Human events reference for ATHEANA (HERA) database description and preliminary user`s manual

    SciTech Connect

    Auflick, J.L.; Hahn, H.A.; Pond, D.J.

    1998-05-27

    The Technique for Human Error Analysis (ATHEANA) is a newly developed human reliability analysis (HRA) methodology that aims to facilitate better representation and integration of human performance into probabilistic risk assessment (PRA) modeling and quantification by analyzing risk-significant operating experience in the context of existing behavioral science models. The fundamental premise of ATHEANA is that error-forcing contexts (EFCs), which refer to combinations of equipment/material conditions and performance shaping factors (PSFs), set up or create the conditions under which unsafe actions (UAs) can occur. Because ATHEANA relies heavily on the analysis of operational events that have already occurred as a mechanism for generating creative thinking about possible EFCs, a database, called the Human Events Reference for ATHEANA (HERA), has been developed to support the methodology. This report documents the initial development efforts for HERA.

  6. Human Events Reference for ATHEANA (HERA) Database Description and Preliminary User's Manual

    SciTech Connect

    Auflick, J.L.

    1999-08-12

    The Technique for Human Error Analysis (ATHEANA) is a newly developed human reliability analysis (HRA) methodology that aims to facilitate better representation and integration of human performance into probabilistic risk assessment (PRA) modeling and quantification by analyzing risk-significant operating experience in the context of existing behavioral science models. The fundamental premise of ATHEANA is that error forcing contexts (EFCs), which refer to combinations of equipment/material conditions and performance shaping factors (PSFs), set up or create the conditions under which unsafe actions (UAs) can occur. Because ATHEANA relies heavily on the analysis of operational events that have already occurred as a mechanism for generating creative thinking about possible EFCs, a database (db) of analytical operational events, called the Human Events Reference for ATHEANA (HERA), has been developed to support the methodology. This report documents the initial development efforts for HERA.

  7. Exploratory Metabolomic Analyses Reveal Compounds Correlated with Lutein Concentration in Frontal Cortex, Hippocampus, and Occipital Cortex of Human Infant Brain

    PubMed Central

    Lieblein-Boff, Jacqueline C.; Johnson, Elizabeth J.; Kennedy, Adam D.; Lai, Chron-Si; Kuchan, Matthew J.

    2015-01-01

    Lutein is a dietary carotenoid well known for its role as an antioxidant in the macula, and recent reports implicate a role for lutein in cognitive function. Lutein is the dominant carotenoid in both pediatric and geriatric brain tissue. In addition, cognitive function in older adults correlated with macular and postmortem brain lutein concentrations. Furthermore, lutein was found to preferentially accumulate in the infant brain in comparison to other carotenoids that are predominant in diet. While lutein is consistently related to cognitive function, the mechanisms by which lutein may influence cognition are not clear. In an effort to identify potential mechanisms through which lutein might influence neurodevelopment, an exploratory study relating metabolite signatures and lutein was completed. Post-mortem metabolomic analyses were performed on human infant brain tissues in three regions important for learning and memory: the frontal cortex, hippocampus, and occipital cortex. Metabolomic profiles were compared to lutein concentration, and correlations were identified and reported here. A total of 1276 correlations were carried out across all brain regions. Of 427 metabolites analyzed, 257 were metabolites of known identity. Unidentified metabolite correlations (510) were excluded. In addition, moderate correlations with xenobiotic relationships (2) or those driven by single outliers (3) were excluded from further study. Lutein concentrations correlated with lipid pathway metabolites, energy pathway metabolites, brain osmolytes, amino acid neurotransmitters, and the antioxidant homocarnosine. These correlations were often brain region—specific. Revealing relationships between lutein and metabolic pathways may help identify potential candidates on which to complete further analyses and may shed light on important roles of lutein in the human brain during development. PMID:26317757

  8. Exploratory Metabolomic Analyses Reveal Compounds Correlated with Lutein Concentration in Frontal Cortex, Hippocampus, and Occipital Cortex of Human Infant Brain.

    PubMed

    Lieblein-Boff, Jacqueline C; Johnson, Elizabeth J; Kennedy, Adam D; Lai, Chron-Si; Kuchan, Matthew J

    2015-01-01

    Lutein is a dietary carotenoid well known for its role as an antioxidant in the macula, and recent reports implicate a role for lutein in cognitive function. Lutein is the dominant carotenoid in both pediatric and geriatric brain tissue. In addition, cognitive function in older adults correlated with macular and postmortem brain lutein concentrations. Furthermore, lutein was found to preferentially accumulate in the infant brain in comparison to other carotenoids that are predominant in diet. While lutein is consistently related to cognitive function, the mechanisms by which lutein may influence cognition are not clear. In an effort to identify potential mechanisms through which lutein might influence neurodevelopment, an exploratory study relating metabolite signatures and lutein was completed. Post-mortem metabolomic analyses were performed on human infant brain tissues in three regions important for learning and memory: the frontal cortex, hippocampus, and occipital cortex. Metabolomic profiles were compared to lutein concentration, and correlations were identified and reported here. A total of 1276 correlations were carried out across all brain regions. Of 427 metabolites analyzed, 257 were metabolites of known identity. Unidentified metabolite correlations (510) were excluded. In addition, moderate correlations with xenobiotic relationships (2) or those driven by single outliers (3) were excluded from further study. Lutein concentrations correlated with lipid pathway metabolites, energy pathway metabolites, brain osmolytes, amino acid neurotransmitters, and the antioxidant homocarnosine. These correlations were often brain region-specific. Revealing relationships between lutein and metabolic pathways may help identify potential candidates on which to complete further analyses and may shed light on important roles of lutein in the human brain during development.

  9. Significance of Poisson distribution theory in analysing the interaction between human spermatozoa and zona-free hamster oocytes.

    PubMed

    Aitken, R J; Elton, R A

    1984-11-01

    The value of Poisson distribution theory in describing and predicting the nature of sperm-egg interaction in vitro has been investigated using an interspecific in-vitro fertilization system, incorporating zona-free hamster oocytes and human spermatozoa. The frequency distribution of polyspermic oocyte penetrations in 72 experiments exhibited good agreement with the Poisson distribution at all levels of fertilization indicating that each oocyte must be of equal penetrability and that there can be no block to polyspermy in this interspecific system. Poisson distribution theory also accurately described the relationship between oocyte penetration and sperm motility in 50 out of 54 separate experiments spread across 10 serial dilution curves. For each dilution series the shape of the fitted curve was fixed but its location along the x-axis varied from donor to donor. The fixed nature of the relationship between sperm motility and egg penetration enables the results of such in-vitro fertilization experiments to be corrected for the number of motile spermatozoa in the incubation media. On the basis of these findings a protocol is described for assessing the results of the zona-free hamster oocyte penetration assay, which involves analysis of the degree of polyspermy followed by the application of Poisson distribution theory to correct the results to a standard concentration of motile spermatozoa. Changes in the penetrating ability of human spermatozoa after vasectomy and characterization of the degree of inter-ejaculate variation in penetrating potential are two clinical examples of such analyses given in the text. The statistical methods described in this paper should also be of general relevance to the study of fertilization mechanisms, in providing a rationale by which to analyse the quantitative nature of sperm-egg interaction in vitro.

  10. Gene therapy for retinitis pigmentosa caused by MFRP mutations: human phenotype and preliminary proof of concept.

    PubMed

    Dinculescu, Astra; Estreicher, Jackie; Zenteno, Juan C; Aleman, Tomas S; Schwartz, Sharon B; Huang, Wei Chieh; Roman, Alejandro J; Sumaroka, Alexander; Li, Qiuhong; Deng, Wen-Tao; Min, Seok-Hong; Chiodo, Vince A; Neeley, Andy; Liu, Xuan; Shu, Xinhua; Matias-Florentino, Margarita; Buentello-Volante, Beatriz; Boye, Sanford L; Cideciyan, Artur V; Hauswirth, William W; Jacobson, Samuel G

    2012-04-01

    Autosomal recessive retinitis pigmentosa (RP), a heterogeneous group of degenerations of the retina, can be due to mutations in the MFRP (membrane-type frizzled-related protein) gene. A patient with RP with MFRP mutations, one of which is novel and the first splice site mutation reported, was characterized by noninvasive retinal and visual studies. The phenotype, albeit complex, suggested that this retinal degeneration may be a candidate for gene-based therapy. Proof-of-concept studies were performed in the rd6 Mfrp mutant mouse model. The fast-acting tyrosine-capsid mutant AAV8 (Y733F) vector containing the small chicken β-actin promoter driving the wild-type mouse Mfrp gene was used. Subretinal vector delivery on postnatal day 14 prevented retinal degeneration. Treatment rescued rod and cone photoreceptors, as assessed by electroretinography and retinal histology at 2 months of age. This AAV-mediated gene delivery also resulted in robust MFRP expression predominantly in its normal location within the retinal pigment epithelium apical membrane and its microvilli. The clinical features of MFRP-RP and our preliminary data indicating a response to gene therapy in the rd6 mouse suggest that this form of RP is a potential target for gene-based therapy.

  11. Preclinical TSPO Ligand PET to Visualize Human Glioma Xenotransplants: A Preliminary Study

    PubMed Central

    Buck, Jason R.; McKinley, Eliot T.; Fu, Allie; Abel, Ty W.; Thompson, Reid C.; Chambless, Lola; Watchmaker, Jennifer M.; Harty, James P.; Cooper, Michael K.; Manning, H. Charles

    2015-01-01

    Current positron emission tomography (PET) imaging biomarkers for detection of infiltrating gliomas are limited. Translocator protein (TSPO) is a novel and promising biomarker for glioma PET imaging. To validate TSPO as a potential target for molecular imaging of glioma, TSPO expression was assayed in a tumor microarray containing 37 high-grade (III, IV) gliomas. TSPO staining was detected in all tumor specimens. Subsequently, PET imaging was performed with an aryloxyanilide-based TSPO ligand, [18F]PBR06, in primary orthotopic xenograft models of WHO grade III and IV gliomas. Selective uptake of [18F]PBR06 in engrafted tumor was measured. Furthermore, PET imaging with [18F]PBR06 demonstrated infiltrative glioma growth that was undetectable by traditional magnetic resonance imaging (MRI). Preliminary PET with [18F]PBR06 demonstrated a preferential tumor-to-normal background ratio in comparison to 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG). These results suggest that TSPO PET imaging with such high-affinity radiotracers may represent a novel strategy to characterize distinct molecular features of glioma growth, as well as better define the extent of glioma infiltration for therapeutic purposes. PMID:26517124

  12. Gene Therapy for Retinitis Pigmentosa Caused by MFRP Mutations: Human Phenotype and Preliminary Proof of Concept

    PubMed Central

    Dinculescu, Astra; Estreicher, Jackie; Zenteno, Juan C.; Aleman, Tomas S.; Schwartz, Sharon B.; Huang, Wei Chieh; Roman, Alejandro J.; Sumaroka, Alexander; Li, Qiuhong; Deng, Wen-Tao; Min, Seok-Hong; Chiodo, Vince A.; Neeley, Andy; Liu, Xuan; Shu, Xinhua; Matias-Florentino, Margarita; Buentello-Volante, Beatriz; Boye, Sanford L.; Cideciyan, Artur V.

    2011-01-01

    Abstract Autosomal recessive retinitis pigmentosa (RP), a heterogeneous group of degenerations of the retina, can be due to mutations in the MFRP (membrane-type frizzled-related protein) gene. A patient with RP with MFRP mutations, one of which is novel and the first splice site mutation reported, was characterized by noninvasive retinal and visual studies. The phenotype, albeit complex, suggested that this retinal degeneration may be a candidate for gene-based therapy. Proof-of-concept studies were performed in the rd6 Mfrp mutant mouse model. The fast-acting tyrosine-capsid mutant AAV8 (Y733F) vector containing the small chicken β-actin promoter driving the wild-type mouse Mfrp gene was used. Subretinal vector delivery on postnatal day 14 prevented retinal degeneration. Treatment rescued rod and cone photoreceptors, as assessed by electroretinography and retinal histology at 2 months of age. This AAV-mediated gene delivery also resulted in robust MFRP expression predominantly in its normal location within the retinal pigment epithelium apical membrane and its microvilli. The clinical features of MFRP-RP and our preliminary data indicating a response to gene therapy in the rd6 mouse suggest that this form of RP is a potential target for gene-based therapy. PMID:22142163

  13. Effects of refrigeration on the bactericidal activity of human milk: a preliminary study.

    PubMed

    Martínez-Costa, Cecilia; Silvestre, María Dolores; López, María Carmen; Plaza, Auxiliadora; Miranda, María; Guijarro, Raquel

    2007-08-01

    This study analyzed the bactericidal activity of human milk and how it is influenced by refrigerated storage. Nine samples of mature human milk were collected and divided into 3 aliquots. One was analyzed immediately, and the other 2 were refrigerated at 4 degrees C to 6 degrees C for 48 and 72 hours, respectively. All of the fresh samples exhibited bactericidal activity with an average value of 83.47% +/- 18.37%. Refrigeration for 48 hours did not cause significant modifications, whereas storage beyond 72 hours significantly lowered the degree of bacteriolysis versus fresh milk. In conclusion, human milk possesses bactericidal activity that remains stable during the first 48 hours of refrigerated storage, but it is significantly reduced beyond 72 hours.

  14. Expression, purification, crystallization and preliminary crystallographic analysis of human Rad GTPase

    SciTech Connect

    Yanuar, Arry; Sakurai, Shigeru; Kitano, Ken; Hakoshima, Toshio

    2005-11-01

    Human Rad has been crystallized. A diffraction data set was collected to a resolution of 1.8 Å. Human Rad is a new member of the Ras GTPase superfamily and is overexpressed in human skeletal muscle of individuals with type II diabetes. The GTPase core domain was overexpressed in Escherichia coli and purified for crystallization. Crystals were obtained at 293 K by vapour diffusion using a crystallization robot. The crystals were found to belong to space group P2{sub 1}, with unit-cell parameters a = 52.2, b = 58.6, c = 53.4 Å, β = 97.9°, and contained two Rad molecules in the crystallographic asymmetric unit. A diffraction data set was collected to a resolution of 1.8 Å using synchrotron radiation at SPring-8.

  15. What Happened, and Why: Toward an Understanding of Human Error Based on Automated Analyses of Incident Reports. Volume 1

    NASA Technical Reports Server (NTRS)

    Maille, Nicolas P.; Statler, Irving C.; Ferryman, Thomas A.; Rosenthal, Loren; Shafto, Michael G.; Statler, Irving C.

    2006-01-01

    The objective of the Aviation System Monitoring and Modeling (ASMM) project of NASA s Aviation Safety and Security Program was to develop technologies that will enable proactive management of safety risk, which entails identifying the precursor events and conditions that foreshadow most accidents. This presents a particular challenge in the aviation system where people are key components and human error is frequently cited as a major contributing factor or cause of incidents and accidents. In the aviation "world", information about what happened can be extracted from quantitative data sources, but the experiential account of the incident reporter is the best available source of information about why an incident happened. This report describes a conceptual model and an approach to automated analyses of textual data sources for the subjective perspective of the reporter of the incident to aid in understanding why an incident occurred. It explores a first-generation process for routinely searching large databases of textual reports of aviation incident or accidents, and reliably analyzing them for causal factors of human behavior (the why of an incident). We have defined a generic structure of information that is postulated to be a sound basis for defining similarities between aviation incidents. Based on this structure, we have introduced the simplifying structure, which we call the Scenario as a pragmatic guide for identifying similarities of what happened based on the objective parameters that define the Context and the Outcome of a Scenario. We believe that it will be possible to design an automated analysis process guided by the structure of the Scenario that will aid aviation-safety experts to understand the systemic issues that are conducive to human error.

  16. Guide to Paperback Translations in the Humanities: A Teacher's Handbook. Preliminary Edition.

    ERIC Educational Resources Information Center

    Raffel, Burton, Comp.

    This teacher's handbook is a guide to available paperback translations of primary material in the humanities field. The selections chosen to be included are recommended by the compiler as the best English translations available. The book is divided into sections chronologically: (1) ancient, to 450 A.D.; (2) medieval, 450-1500 A.D.; (3) early…

  17. Preliminary evidence for an association between LRP-1 genotype and body mass index in humans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The LDL receptor-related protein-1 gene (LRP-1) has been associated with obesity in animal models, but no such association has yet been reported in humans. As data suggest this increase in fat mass may be mediated through a mechanism involving the clearance of plasma triglyceride-rich lipoproteins (...

  18. Human Relations and Community Life in Rural New York State: A Preliminary Report.

    ERIC Educational Resources Information Center

    New York State Legislative Commission on Rural Resources, Albany.

    Trends, strengths and assets, weaknesses and problem areas, goals, and public policy questions in the area of human relations and community life in rural New York state are presented with supporting statistics. Trends considered include rural and elderly rural population increases; suicide, homicide, and domestic violence rate increases; demands…

  19. 76 FR 39399 - Chlorpyrifos Registration Review; Preliminary Human Health Risk Assessment; Notice of Availability

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-06

    ... registration review of chlorpyrifos pursuant to section 3(g) of the Federal Insecticide, Fungicide, and... unreasonable adverse effects on human health or the environment. Chlorpyrifos is an organophosphate insecticide... for the Southern District of New York in Natural Resources Defense Council, et al. v. EPA, No. 10...

  20. Evaluation of the interaction between calcifying nanoparticles and human dental pulp cells: a preliminary investigation.

    PubMed

    Yang, Fang; Zeng, Jinfeng; Zhang, Wei; Sun, Xi; Ling, Junqi

    2010-12-15

    Calcifying nanoparticles (CNPs, previously called nanobacteria) are self-propagating, cultivable macromolecular complexes. Their extraordinary characteristic is that they can aggregate carbonate apatite on their envelope from soluble calcium and phosphorus at physiologic concentrations and display cytotoxic effects on murine and human fibroblast cell lines. The question arises whether CNPs contribute to the degeneration of pulp tissue and thus result in clinically significant human dental pulp stones as nidies. This study evaluates CNPs' effects upon human dental pulp cells (HDPCs, the host cells in pulp tissue). We observed the ultrastructural variation of HDPCs attacked by CNPs. The spatial relationship of HDPCs and CNPs after coculture was also identified by immunofluorescence staining. Furthermore, it was verified by MTT viability assay that CNPs isolated from dental pulp stones exerted cytotoxic effect on HDPCs. Therefore, it could be concluded that the existence of CNPs might interfere with the normal physiologic function of the cells, and that might lead to dental pulp calcification. Elucidation of the cytotoxic characteristics of CNPs may offer a new perspective for understanding the etiology of human dental pulp stones.

  1. Stardust Interstellar Preliminary Examination IV: Scanning Transmission X-Ray Microscopy Analyses of Impact Features in the Stardust Interstellar Dust Collector

    NASA Technical Reports Server (NTRS)

    Butterworth, Anna L.; Westphal, Andrew J.; Frank, David R.; Allen, Carlton C.; Bechtel, Hans A.; Sandford, Scott A.; Tsou, Peter; Zolensky, Michael E.

    2014-01-01

    We report the quantitative characterization by synchrotron soft X-ray spectroscopy of 31 potential impact features in the aerogel capture medium of the Stardust Interstellar Dust Collector. Samples were analyzed in aerogel by acquiring high spatial resolution maps and high energy-resolution spectra of major rock-forming elements Mg, Al, Si, Fe, and others. We developed diagnostic screening tests to reject spacecraft secondary ejecta and terrestrial contaminants from further consideration as interstellar dust candidates. The results support an extraterrestrial origin for three interstellar candidates: I1043,1,30 (Orion) is a 3 pg particle with Mg-spinel, forsterite, and an iron-bearing phase. I1047,1,34 (Hylabrook) is a 4 pg particle comprising an olivine core surrounded by low-density, amorphous Mg-silicate and amorphous Fe, Cr, and Mn phases. I1003,1,40 (Sorok) has the track morphology of a high-speed impact, but contains no detectable residue that is convincingly distinguishable from the background aerogel. Twenty-two samples with an anthropogenic origin were rejected, including four secondary ejecta from impacts on the Stardust spacecraft aft solar panels, nine ejecta from secondary impacts on the Stardust Sample Return Capsule, and nine contaminants lacking evidence of an impact. Other samples in the collection included I1029,1,6, which contained surviving solar system impactor material. Four samples remained ambiguous: I1006,2,18, I1044,2,32, and I1092,2,38 were too dense for analysis, and we did not detect an intact projectile in I1044,3,33. We detected no radiation effects from the synchrotron soft X-ray analyses; however, we recorded the effects of synchrotron hard X-ray radiation on I1043,1,30 and I1047,1,34.

  2. Human-FES Cooperative Control for Wrist Movement: A Preliminary Study

    PubMed Central

    Gui, Kai; Yokoi, Hiroshi; Zhang, Dingguo

    2016-01-01

    Functional electrical stimulation (FES) sometimes applies to patients with partial paralysis, so human voluntary control and FES control both exist. Our study aims to build a cooperative controller to achieve human-FES cooperation. This cooperative controller is formed by a classical FES controller and an impedance controller. The FES controller consists of a back propagation (BP) neural network-based feedforward controller and a PID-based feedback controller. The function of impedance controller is to convert volitional force/torque, which is estimated from a three-stage filter based on EMG, into additional angle. The additional angle can reduce the FES intensity in our cooperative controller, comparing to that in classical FES controller. Some assessment experiments are designed to test the performance of the cooperative controller. PMID:27990243

  3. Enhanced Contaminated Human Remains Pouch: initial development and preliminary performance assessments

    SciTech Connect

    Iseli, A.M.; Kwen, H.D.; Ul-Alam, M.; Balasubramanian, M.; Rajagopalan, S.

    2011-11-07

    The objective is to produce a proof of concept prototype Enhanced Contaminated Human Remains Pouch (ECHRP) with self-decontamination capability to provide increased protection to emergency response personnel. The key objective was to decrease the concentration of toxic chemicals through the use of an absorbent and reactive nanocellulose liner. Additionally, nanomaterials with biocidal properties were developed and tested as a 'stand-alone' treatment. The setting was a private company research laboratory. The main outcome measures were production of a functional prototype. A functional prototype capable of mitigating the threats due to sulfur mustard, Soman, and a large variety of liquid and vapor toxic industrial chemicals was produced. Stand-alone biocidal treatment efficacy was validated. The ECHRP provides superior protection from both chemical and biological hazards to various emergency response personnel and human remains handlers.

  4. Implementing evidence-based practice in human service organizations: preliminary lessons from the frontlines.

    PubMed

    Austin, Michael J; Claassen, Jennette

    2008-01-01

    Evidence-based practice (EBP) involves the integration of the expertise of individual practitioners with the best available evidence within the context of values and expectations of clients. Little is known about the implementation of evidence-based practice in the human services. This article is based on a comprehensive search of the literature related to the organizational factors needed to introduce EBP into a human service agency, tools for assessing organizational readiness for EBP, and lessons learned from the current implementation efforts. Three approaches to implementing EBP are investigated: the micro (increasing worker skills), macro (strengthening systems and structures), and the combination (focusing on both aspects). Conclusions and recommendations are drawn from the literature review and framed in the form of a tool for assessing organizational readiness for EBP implementation.

  5. Preliminary neutron diffraction analysis of challenging human manganese superoxide dismutase crystals.

    PubMed

    Azadmanesh, Jahaun; Trickel, Scott R; Weiss, Kevin L; Coates, Leighton; Borgstahl, Gloria E O

    2017-04-01

    Superoxide dismutases (SODs) are enzymes that protect against oxidative stress by dismutation of superoxide into oxygen and hydrogen peroxide through cyclic reduction and oxidation of the active-site metal. The complete enzymatic mechanisms of SODs are unknown since data on the positions of hydrogen are limited. Here, methods are presented for large crystal growth and neutron data collection of human manganese SOD (MnSOD) using perdeuteration and the MaNDi beamline at Oak Ridge National Laboratory. The crystal from which the human MnSOD data set was obtained is the crystal with the largest unit-cell edge (240 Å) from which data have been collected via neutron diffraction to sufficient resolution (2.30 Å) where hydrogen positions can be observed.

  6. Human-FES Cooperative Control for Wrist Movement: A Preliminary Study.

    PubMed

    Gui, Kai; Yokoi, Hiroshi; Zhang, Dingguo

    2016-06-13

    Functional electrical stimulation (FES) sometimes applies to patients with partial paralysis, so human voluntary control and FES control both exist. Our study aims to build a cooperative controller to achieve human-FES cooperation. This cooperative controller is formed by a classical FES controller and an impedance controller. The FES controller consists of a back propagation (BP) neural network-based feedforward controller and a PID-based feedback controller. The function of impedance controller is to convert volitional force/torque, which is estimated from a three-stage filter based on EMG, into additional angle. The additional angle can reduce the FES intensity in our cooperative controller, comparing to that in classical FES controller. Some assessment experiments are designed to test the performance of the cooperative controller.

  7. Characterization of the Human Proteomic Response to Hydrocodone: A Preliminary Study

    DTIC Science & Technology

    2013-03-01

    hippocampus and striatum after exposure to morphine , lnt J Mol Med 18 (2006) 775-784. [50] Z.H. Wen, G.J. Wu, L.C. Hsu, W.F. Chen, J.Y. Chen, H.A...McMillin, F.M. Urry, Simultaneous determination of codeine, morphine , hydrocodone, hydromorphone, oxycodone, and 6-acetylmorphine in urine, serum...Simultaneous assay of morphine , morphine -3-glucuronide and morphine -6-glucuronide in human plasma using normal-phase liquid chromatography-tandem

  8. The impact of Body Worlds on adult visitors' knowledge on human anatomy: A preliminary study.

    PubMed

    Fonseca, Guilherme R B C; Finn, Gabrielle M

    2016-05-01

    Body Worlds is an anatomical exhibition that shows human remains to the public. It has been considered controversial since it raises ethical tensions and issues. However, organizers and supporters of Body Worlds have claimed the exhibition is intended to promote visitors' understanding over the human body. Despite these claims, no studies were found that support or refute the hypothesis that a visit to Body Worlds increases the public's objective knowledge on human anatomy. Consequently, the objective of this study was to determine the impact of Body Worlds on anatomical knowledge. We constructed and delivered a questionnaire to both a previsit random sample and a postvisit random sample of visitors of Body Worlds' event Facets of Life, in Berlin. The questionnaire was available in both English and German languages and contained (a) basic sociodemographic questions and (b) a valid and reliable anatomy quiz. The quiz consisted of 16 multiple-choice questions that assessed the ability to identify the location of major anatomical structures on the human body. Average scores achieved on the quiz by the postvisit sample (X¯= 9.08, s = 2.48, n = 164) were significantly higher (unpaired t = 3.3957, P = 0.0008) than those achieved by the previsit sample (X¯= 8.11, s = 2.69, n = 167). Our results suggest that a visit to Body Worlds' event Facets of Life may have a beneficial effect in anatomical knowledge. However, further studies with better empirical designs and fewer limitations are needed to confirm our results.

  9. [Preliminary establishment of transplanted human chronic myeloid leukemia model in nude mice].

    PubMed

    Li, Xian-Min; Ding, Xin; Zhang, Long-Zhen; Cen, Jian-Nong; Chen, Zi-Xing

    2011-12-01

    Chronic myeloid leukemia (CML) is a malignant clonal disease derived from hematopoietic stem cells. CML stem cells were thought to be the root which could lead disease development and ultimately rapid change. However, a stable animal model for studying the characteristics of CML stem cells is currently lacking. This study was aimed to establish a transplanted human CML nude-mice model to further explore the biological behavior of CML stem cells in vivo, and to enrich CML stem cells in nude mice by series transplantation. The 4 - 6 weeks old BALB/c nude mice pretreated by splenectomy (S), cytoxan intraperitoneal injection (C) and sublethal irradiation (I) were transplanted intravenously with (5 - 7) × 10(7) of bone marrow mononuclear cells from CML patients in chronic phase. Alternatively, 4 - 6 weeks old BALB/c nude mice pretreated by lethal irradiation were transplanted intravenously with 5 × 10(6) homologous bone marrow cells of BALB/c nude mice together with (5 - 7) × 10(7) of bone marrow mononuclear cells from CML patients in chronic phase simultaneously. The leukemic cells engrafted and infiltrated in organs and bone marrow of the mice were tracked by reverse transcription-polymerase chain reaction (RT-PCR), plastic-embedded biopsy and flow cytometry. The results of these two methods were compared. The results showed that human CML cells engrafted and infiltrating into the bone marrow of two nude mice pretreated with SCI could be detected. In spite of the low successful rate, results suggested the feasibility of this method by using BALB/c nude mice as a human CML animal model. In contrast, in nude mice pretreated by the lethal dose irradiation, CML cells in the bone marrow could not be found. It is concluded that human bone marrow CML cells can results in leukemia in nude mice pretreated by SCI. Thus this study provides a new strategy for establishment of CML animal models which deserves further elaboration.

  10. Normal human mitral valve proteome: A preliminary investigation by gel-based and gel-free proteomic approaches.

    PubMed

    Brioschi, Maura; Baetta, Roberta; Ghilardi, Stefania; Gianazza, Erica; Guarino, Anna; Parolari, Alessandro; Polvani, Gianluca; Tremoli, Elena; Banfi, Cristina

    2016-10-01

    The mitral valve is a highly complex structure which regulates blood flow from the left atrium to the left ventricle (LV) avoiding a significant forward gradient during diastole or regurgitation during systole. The integrity of the mitral valve is also essential for the maintenance of normal LV size, geometry, and function. Significant advances in the comprehension of the biological, functional, and mechanical behavior of the mitral valve have recently been made. However, current knowledge of protein components in the normal human mitral valve is still limited and complicated by the low cellularity of this tissue and the presence of high abundant proteins from the extracellular matrix. We employed here an integrated proteomic approach to analyse the protein composition of the normal human mitral valve and reported confident identification of 422 proteins, some of which have not been previously described in this tissue. In particular, we described the ability of pre-MS separation technique based on liquid-phase IEF and SDS-PAGE to identify the largest number of proteins. We also demonstrated that some of these proteins, e.g. αB-Crystallin, septin-11, four-and-a-half LIM domains protein 1, and dermatopontin, are synthesised by interstitial cells isolated from human mitral valves. These initial results provide a valuable basis for future studies aimed at analysing in depth the mitral valve protein composition and at investigating potential pathogenetic molecular mechanisms. Data are available via ProteomeXchange with identifier PXD004397.

  11. Facilitating myoelectric-control with transcranial direct current stimulation: a preliminary study in healthy humans

    PubMed Central

    2014-01-01

    during 'proportional EMG control’ when compared to M1 anodal and sham tDCS. Conclusions The preliminary results from healthy subjects showed specific, and at least partially antagonistic effects, of M1 and cerebellar anodal tDCS on motor performance during myoelectric control. These results are encouraging, but further studies are necessary to better define how tDCS over particular regions of the cerebellum may facilitate learning of myoelectric control for brain machine interfaces. PMID:24507410

  12. Modeling EEG fractal dimension changes in wake and drowsy states in humans--a preliminary study.

    PubMed

    Bojić, Tijana; Vuckovic, Aleksandra; Kalauzi, Aleksandar

    2010-01-21

    Aim of this preliminary study was to examine and compare topographic distribution of Higuchi's fractal dimension (FD, measure of signal complexity) of EEG signals between states of relaxed wakefulness and drowsiness, as well as their FD differences. The experiments were performed on 10 healthy individuals using a fourteen-channel montage. An explanation is offered on the causes of the detected FD changes. FD values of 60s records belonging to wake (Hori's stage 1) and drowsy (Hori's stages 2-4) states were calculated for each channel and each subject. In 136 out of 140 epochs an increase in FD was obtained. Relationship between signal FD and its relative alpha amplitude was mathematically modeled and we quantitatively demonstrated that the increase in FD was predominantly due to a reduction in alpha activity. The model was generalized to include other EEG oscillations. By averaging FD values for each channel across 10 subjects, four clusters (O2O1; T6P4T5P3; C3F3F4C4F8F7; T4T3) for the wake and two clusters (O2O1P3T6P4T5; C3C4F4F3F8T4T3F7) for the drowsy state were statistically verified. Topographic distribution of FD values in wakefulness showed a lateral symmetry and a partial fronto-occipital gradient. In drowsiness, a reduction in the number of clusters was detected, due to regrouping of channels T3, T4, O1 and O2. Topographic distribution of absolute FD differences revealed largest values at F7, O1 and F3. Reorganization of channel clusters showed that regionalized brain activity, specific for wakefulness, became more global by entering into drowsiness. Since the global increase in FD during wake-to-drowsy transition correlated with the decrease of alpha power, we inferred that increase of EEG complexity may not necessarily be an index of brain activation.

  13. Preliminary experiments on dynamic biology of micro-organisms to avoid any specific full-blown syndrome on humans

    NASA Astrophysics Data System (ADS)

    Meer, Sneer

    2002-06-01

    The aim of this paper is to apply an efficient system to detect, identify and quicken suppression of any dangerous micro-organism which threatens the health of the human body in any form. It is well known that some specimens of this kind of possess a specific energy related to their speed of division, toxin emissions and high-powered interaction with human and animal cells which have the capacity to provide certain deadly full-blown syndromes. Many problems relating to the above-mentioned properties have not been clarified to date, and it is vital to find a rapid and valid reply as soon as possible. Inter-disciplinary sciences directed us to start some experiments to solve such problems, considering that the human body is dotted with a multiple interactive system of energy release, a fact which can explain the source of the micro-organism's energy also, for their necessity to manifest their deadly pathology. From practical preliminary experiments with some micro-mechanical systems using light-microscopy, connected to video TV Recorder System, one obtains optical enlarged TV images of certain processes which indicated the right way towards our crucial target; ie: the preparation of safe vaccines and safe medicines. This will constitute a basic system to a void deadly manifestations of dangerous micro-organisms and/or even regular infections on earth and in space, a system which will probably be applied at the ISS Space Station and other future actions in space in long and very long flights. We look forward to applying this system of dynamic biology towards preparation of a real and valid vaccine(s) against HIV virus on AIDS diseases.

  14. Crystallization and preliminary X-ray diffraction analyses of the TIR domains of three TIR-NB-LRR proteins that are involved in disease resistance in Arabidopsis thaliana.

    PubMed

    Wan, Li; Zhang, Xiaoxiao; Williams, Simon J; Ve, Thomas; Bernoux, Maud; Sohn, Kee Hoon; Jones, Jonathan D G; Dodds, Peter N; Kobe, Bostjan

    2013-11-01

    The Toll/interleukin-1 receptor (TIR) domain is a protein-protein interaction domain that is found in both animal and plant immune receptors. The N-terminal TIR domain from the nucleotide-binding (NB)-leucine-rich repeat (LRR) class of plant disease-resistance (R) proteins has been shown to play an important role in defence signalling. Recently, the crystal structure of the TIR domain from flax R protein L6 was determined and this structure, combined with functional studies, demonstrated that TIR-domain homodimerization is a requirement for function of the R protein L6. To advance the molecular understanding of the function of TIR domains in R-protein signalling, the protein expression, purification, crystallization and X-ray diffraction analyses of the TIR domains of the Arabidopsis thaliana R proteins RPS4 (resistance to Pseudomonas syringae 4) and RRS1 (resistance to Ralstonia solanacearum 1) and the resistance-like protein SNC1 (suppressor of npr1-1, constitutive 1) are reported here. RPS4 and RRS1 function cooperatively as a dual resistance-protein system that prevents infection by three distinct pathogens. SNC1 is implicated in resistance pathways in Arabidopsis and is believed to be involved in transcriptional regulation through its interaction with the transcriptional corepressor TPR1 (Topless-related 1). The TIR domains of all three proteins have successfully been expressed and purified as soluble proteins in Escherichia coli. Plate-like crystals of the RPS4 TIR domain were obtained using PEG 3350 as a precipitant; they diffracted X-rays to 2.05 Å resolution, had the symmetry of space group P1 and analysis of the Matthews coefficient suggested that there were four molecules per asymmetric unit. Tetragonal crystals of the RRS1 TIR domain were obtained using ammonium sulfate as a precipitant; they diffracted X-rays to 1.75 Å resolution, had the symmetry of space group P4(1)2(1)2 or P4(3)2(1)2 and were most likely to contain one molecule per asymmetric

  15. Comparison of macronutrient contents in human milk measured using mid-infrared human milk analyser in a field study vs. chemical reference methods.

    PubMed

    Zhu, Mei; Yang, Zhenyu; Ren, Yiping; Duan, Yifan; Gao, Huiyu; Liu, Biao; Ye, Wenhui; Wang, Jie; Yin, Shian

    2017-01-01

    Macronutrient contents in human milk are the common basis for estimating these nutrient requirements for both infants and lactating women. A mid-infrared human milk analyser (HMA, Miris, Sweden) was recently developed for determining macronutrient levels. The purpose of the study is to compare the accuracy and precision of HMA method with fresh milk samples in the field studies with chemical methods with frozen samples in the lab. Full breast milk was collected using electric pumps and fresh milk was analyzed in the field studies using HMA. All human milk samples were thawed and analyzed with chemical reference methods in the lab. The protein, fat and total solid levels were significantly correlated between the two methods and the correlation coefficient was 0.88, 0.93 and 0.78, respectively (p  <  0.001). The mean protein content was significantly lower and the mean fat level was significantly greater when measured using HMA method (1.0 g 100 mL(-1) vs 1.2 g 100 mL(-1) and 3. 7 g 100 mL(-1) vs 3.2 g 100 mL(-1) , respectively, p  <  0.001). Thus, linear recalibration could be used to improve mean estimation for both protein and fat. There was no significant correlation for lactose between the two methods (p  >  0.05). There was no statistically significant difference in the mean total solid concentration (12.2 g 100 mL(-1) vs 12.3 g 100 mL(-1) , p  >  0.05). Overall, HMA might be used to analyze macronutrients in fresh human milk with acceptable accuracy and precision after recalibrating fat and protein levels of field samples.

  16. The application of traditional and geometric morphometric analyses for forensic quantification of sexual dimorphism: preliminary investigations in a Western Australian population.

    PubMed

    Franklin, Daniel; Cardini, Andrea; Flavel, Ambika; Kuliukas, Algis

    2012-07-01

    A current limitation of forensic practice in Western Australia is a lack of contemporary population-specific standards for biological profiling; this directly relates to the unavailability of documented human skeletal collections. With rapidly advancing technology, however, it is now possible to acquire accurate skeletal measurements from 3D scans contained in medical databases. The purpose of the present study, therefore, is to explore the accuracy of using cranial form to predict sex in adult Australians. Both traditional and geometric morphometric methods are applied to data derived from 3D landmarks acquired in CT-reconstructed crania. The sample comprises multi-detector computed tomography scans of 200 adult individuals; following 3D volume rendering, 46 anatomical landmarks are acquired using OsiriX (version 3.9). Centroid size and shape (first 20 PCs of the Procrustes coordinates) and the inter-landmark (ILD) distances between all possible pairs of landmarks are then calculated. Sex classification effectiveness of the 3D multivariate descriptors of size and shape and selected ILD measurements are assessed and compared; robustness of findings is explored using resampling statistics. Cranial shape and size and the ILD measurements are sexually dimorphic and explain 3.2 to 54.3 % of sample variance; sex classification accuracy is 83.5-88.0 %. Sex estimation using 3D shape appears to have some advantages compared to approaches using size measurements. We have, however, identified a simple and biologically meaningful single non-traditional linear measurement (glabella-zygion) that classifies Western Australian individuals according to sex with a high degree of expected accuracy (87.5-88 %).

  17. Mouse and Human Genetic Analyses Associate Kalirin with Ventral Striatal Activation during Impulsivity and with Alcohol Misuse

    PubMed Central

    Peña-Oliver, Yolanda; Carvalho, Fabiana M.; Sanchez-Roige, Sandra; Quinlan, Erin B.; Jia, Tianye; Walker-Tilley, Tom; Rulten, Stuart L.; Pearl, Frances M. G.; Banaschewski, Tobias; Barker, Gareth J.; Bokde, Arun L. W.; Büchel, Christian; Conrod, Patricia J.; Flor, Herta; Gallinat, Jürgen; Garavan, Hugh; Heinz, Andreas; Gowland, Penny; Paillere Martinot, Marie-Laure; Paus, Tomáš; Rietschel, Marcella; Robbins, Trevor W.; Smolka, Michael N.; Schumann, Gunter; Stephens, David N.

    2016-01-01

    Impulsivity is associated with a spectrum of psychiatric disorders including drug addiction. To investigate genetic associations with impulsivity and initiation of drug taking, we took a two-step approach. First, we identified genes whose expression level in prefrontal cortex, striatum and accumbens were associated with impulsive behavior in the 5-choice serial reaction time task across 10 BXD recombinant inbred (BXD RI) mouse strains and their progenitor C57BL/6J and DBA2/J strains. Behavioral data were correlated with regional gene expression using GeneNetwork (www.genenetwork.org), to identify 44 genes whose probability of association with impulsivity exceeded a false discovery rate of < 0.05. We then interrogated the IMAGEN database of 1423 adolescents for potential associations of SNPs in human homologs of those genes identified in the mouse study, with brain activation during impulsive performance in the Monetary Incentive Delay task, and with novelty seeking scores from the Temperament and Character Inventory, as well as alcohol experience. There was a significant overall association between the human homologs of impulsivity-related genes and percentage of premature responses in the MID task and with fMRI BOLD-response in ventral striatum (VS) during reward anticipation. In contrast, no significant association was found between the polygenic scores and anterior cingulate cortex activation. Univariate association analyses revealed that the G allele (major) of the intronic SNP rs6438839 in the KALRN gene was significantly associated with increased VS activation. Additionally, the A-allele (minor) of KALRN intronic SNP rs4634050, belonging to the same haplotype block, was associated with increased frequency of binge drinking. PMID:27092175

  18. Proteomic Analyses Reveal Common Promiscuous Patterns of Cell Surface Proteins on Human Embryonic Stem Cells and Sperms

    PubMed Central

    Gu, Bin; Zhang, Jiarong; Wu, Ying; Zhang, Xinzong; Tan, Zhou; Lin, Yuanji; Huang, Xiao; Chen, Liangbiao; Yao, Kangshou; Zhang, Ming

    2011-01-01

    Background It has long been proposed that early embryos and reproductive organs exhibit similar gene expression profiles. However, whether this similarity is propagated to the protein level remains largely unknown. We have previously characterised the promiscuous expression pattern of cell surface proteins on mouse embryonic stem (mES) cells. As cell surface proteins also play critical functions in human embryonic stem (hES) cells and germ cells, it is important to reveal whether a promiscuous pattern of cell surface proteins also exists for these cells. Methods and Principal Findings Surface proteins of hES cells and human mature sperms (hSperms) were purified by biotin labelling and subjected to proteomic analyses. More than 1000 transmembrane or secreted cell surface proteins were identified on the two cell types, respectively. Proteins from both cell types covered a large variety of functional categories including signal transduction, adhesion and transporting. Moreover, both cell types promiscuously expressed a wide variety of tissue specific surface proteins, and some surface proteins were heterogeneously expressed. Conclusions/Significance Our findings indicate that the promiscuous expression of functional and tissue specific cell surface proteins may be a common pattern in embryonic stem cells and germ cells. The conservation of gene expression patterns between early embryonic cells and reproductive cells is propagated to the protein level. These results have deep implications for the cell surface signature characterisation of pluripotent stem cells and germ cells and may lead the way to a new area of study, i.e., the functional significance of promiscuous gene expression in pluripotent and germ cells. PMID:21559292

  19. Hindlimb muscle architecture in non-human great apes and a comparison of methods for analysing inter-species variation

    PubMed Central

    Myatt, Julia P; Crompton, Robin H; Thorpe, Susannah K S

    2011-01-01

    By relating an animal's morphology to its functional role and the behaviours performed, we can further develop our understanding of the selective factors and constraints acting on the adaptations of great apes. Comparison of muscle architecture between different ape species, however, is difficult because only small sample sizes are ever available. Further, such samples are often comprised of different age–sex classes, so studies have to rely on scaling techniques to remove body mass differences. However, the reliability of such scaling techniques has been questioned. As datasets increase in size, more reliable statistical analysis may eventually become possible. Here we employ geometric and allometric scaling techniques, and ancovas (a form of general linear model, GLM) to highlight and explore the different methods available for comparing functional morphology in the non-human great apes. Our results underline the importance of regressing data against a suitable body size variable to ascertain the relationship (geometric or allometric) and of choosing appropriate exponents by which to scale data. ancova models, while likely to be more robust than scaling for species comparisons when sample sizes are high, suffer from reduced power when sample sizes are low. Therefore, until sample sizes are radically increased it is preferable to include scaling analyses along with ancovas in data exploration. Overall, the results obtained from the different methods show little significant variation, whether in muscle belly mass, fascicle length or physiological cross-sectional area between the different species. This may reflect relatively close evolutionary relationships of the non-human great apes; a universal influence on morphology of generalised orthograde locomotor behaviours or, quite likely, both. PMID:21507000

  20. Comparative analyses of the complete mitochondrial genomes of Ascaris lumbricoides and Ascaris suum from humans and pigs.

    PubMed

    Liu, Guo-Hua; Wu, Chang-Yi; Song, Hui-Qun; Wei, Shu-Jun; Xu, Min-Jun; Lin, Rui-Qing; Zhao, Guang-Hui; Huang, Si-Yang; Zhu, Xing-Quan

    2012-01-15

    Ascaris lumbricoides and Ascaris suum are parasitic nematodes living in the small intestine of humans and pigs, and can cause the disease ascariasis. For long, there has been controversy as to whether the two ascaridoid taxa represent the same species due to their significant resemblances in morphology. However, the complete mitochondrial (mt) genome data have been lacking for A. lumbricoides in spite of human and animal health significance and socio-economic impact globally of these parasites. In the present study, we sequenced the complete mt genomes of A. lumbricoides and A. suum (China isolate), which was 14,303 bp and 14,311 bp in size, respectively. The identity of the mt genomes was 98.1% between A. lumbricoides and A. suum (China isolate), and 98.5% between A. suum (China isolate) and A. suum (USA isolate). Both genomes are circular, and consist of 36 genes, including 12 genes for proteins, 2 genes for rRNA and 22 genes for tRNA, which are consistent with that of all other species of ascaridoid studied to date. All genes are transcribed in the same direction and have a nucleotide composition high in A and T (71.7% for A. lumbricoides and 71.8% for A. suum). The AT bias had a significant effect on both the codon usage pattern and amino acid composition of proteins. Phylogenetic analyses of A. lumbricoides and A. suum using concatenated amino acid sequences of 12 protein-coding genes, with three different computational algorithms (Bayesian analysis, maximum likelihood and maximum parsimony) all clustered in a clade with high statistical support, indicating that A. lumbricoides and A. suum was very closely related. These mt genome data and the results provide some additional genetic evidence that A. lumbricoides and A. suum may represent the same species. The mt genome data presented in this study are also useful novel markers for studying the molecular epidemiology and population genetics of Ascaris.

  1. NOTE: A preliminary study for non-invasive quantification of manganese in human hand bones

    NASA Astrophysics Data System (ADS)

    Aslam; Pejović-Milić, A.; Chettle, D. R.; McNeill, F. E.; Pysklywec, M. W.; Oudyk, J.

    2008-10-01

    Manganese (Mn) is a nutrient essential for regulating neurological and skeletal functions in the human body, but it is also toxic when humans are excessively exposed to Mn. Blood (or serum/plasma) and other body fluids reflect only the most recent exposure and rapidly return to within normal ranges, even when there has been a temporary excursion in response to exposure. In this context, we have been developing a non-invasive measurement of Mn stored in bone, using in vivo neutron activation analysis. Following feasibility studies, a first pilot study, using neutron activation analysis to measure Mn in the bones of the hand of ten healthy male human subjects, was conducted with the approval of the concerned research ethics boards. The participants of this study had no known history of exposure to Mn. Two volunteers were excluded from this study due to technical problems with their measurements. The inverse variance weighted mean value of Mn/Ca for the participants of this study is 0.12 ± 0.68 µg Mn/g Ca which is comparable within uncertainties with the estimated range of 0.16 0.78 µg Mn/g Ca and mean value of 0.63 ± 0.30 µg Mn/g Ca derived from cadaver data. It is recommended to investigate the use of the diagnostic technique for in vivo measurements of workers exposed occupationally to excessive amounts of Mn who could develop many-fold increased levels of Mn in bones as demonstrated through various animal studies. The technique needs further development to improve the precision of in vivo measurements in the non-exposed population.

  2. Characterization of the Human Proteomic Response to Hydrocodone: A Preliminary Study

    DTIC Science & Technology

    2012-03-01

    analysis of rat cerebral cortex, hippocampus and striatum after exposure to morphine , In! J Mol Med 18 (2006) 775-784. [50] Z.H. Wen, G.J. Wu, L.C...Coles, M.M. Kushnir, G.J. Nelson, G.A. McMillin, F.M. Urry, Simultaneous determination of codeine, morphine , hydrocodone, hydromorphone, oxycodone...Jiang, M. Wehling , J.D. Hulse, P.P. Lin, Simultaneous assay of morphine , morphine -3-glucuronide and morphine -6-glucuronide in human plasma using

  3. Characterization of the Human Proteomic Response to Hydrocodone: A Preliminary Study

    DTIC Science & Technology

    2014-05-01

    cortex, hippocampus and striatum after exposure to morphine , Int J Mol Med 18 (2006) 775-784. [50] Z.H. Wen, G.J. Wu, L.C. Hsu, W.F. Chen, J.Y. Chen...Nelson, G.A. McMillin, F.M. Urry, Simultaneous determination of codeine, morphine , hydrocodone, hydromorphone, oxycodone, and 6-acetylmorphine in...Hulse, P.P. Lin, Simultaneous assay of morphine , morphine -3-glucuronide and morphine -6-glucuronide in human plasma using normal-phase liquid

  4. Fukushima nuclear accident: preliminary assessment of the risks to non-human biota.

    PubMed

    Aliyu, Abubakar Sadiq; Ramli, Ahmad Termizi; Garba, Nuraddeen Nasiru; Saleh, Muneer Aziz; Gabdo, Hamman Tukur; Liman, Muhammad Sanusi

    2015-02-01

    This study assesses the 'radio-ecological' impacts of Fukushima nuclear accident on non-human biota using the ERICA Tool, which adopts an internationally verified methodology. The paper estimates the impacts of the accident on terrestrial and marine biota based on the environmental data reported in literature for Japan, China, South Korea and the USA. Discernible impacts have been detected in the marine biota around Fukushima Daiichi nuclear power plant. This study confirms that the Fukushima accident had caused heavier damage to marine bionts compared with terrestrial flora and fauna, in Japan.

  5. Technical note: 3D from standard digital photography of human crania-a preliminary assessment.

    PubMed

    Katz, David; Friess, Martin

    2014-05-01

    This study assessed three-dimensional (3D) photogrammetry as a tool for capturing and quantifying human skull morphology. While virtual reconstruction with 3D surface scanning technology has become an accepted part of the paleoanthropologist's tool kit, recent advances in 3D photogrammetry make it a potential alternative to dedicated surface scanners. The principal advantages of photogrammetry are more rapid raw data collection, simplicity and portability of setup, and reduced equipment costs. We tested the precision and repeatability of 3D photogrammetry by comparing digital models of human crania reconstructed from conventional, 2D digital photographs to those generated using a 3D surface scanner. Overall, the photogrammetry and scanner meshes showed low degrees of deviation from one another. Surface area estimates derived from photogrammetry models tended to be slightly larger. Landmark configurations generally did not cluster together based upon whether the reconstruction was created with photogrammetry or surface scanning technology. Average deviations of landmark coordinates recorded on photogrammetry models were within the generally allowable range of error in osteometry. Thus, while dependent upon the needs of the particular research project, 3D photogrammetry appears to be a suitable, lower-cost alternative to 3D imaging and scanning options.

  6. Crystallization and preliminary crystallographic analysis of human Atg4B–LC3 complex

    SciTech Connect

    Satoo, Kenji; Suzuki, Nobuo N.; Fujioka, Yuko; Mizushima, Noboru; Ohsumi, Yoshinori; Inagaki, Fuyuhiko

    2007-02-01

    Human Atg4B and LC3 were expressed, purified and crystallized as a complex. Diffraction data were collected to a resolution of 1.9 Å. The reversible modification of Atg8 with phosphatidylethanolamine (PE) is crucial for autophagy, the bulk degradation process of cytoplasmic components by the vacuolar/lysosomal system. Atg4 is a cysteine protease that is responsible for the processing and deconjugation of Atg8. Human Atg4B (HsAtg4B; a mammalian orthologue of yeast Atg4) and LC3 (a mammalian orthologue of yeast Atg8) were expressed and purified and two complexes, one consisting of HsAtg4B(1–354) and LC3(1–120) (complex I; the product complex) and the other consisting of HsAtg4B(1–354) and LC3(1–124) (complex II; the substrate complex), were crystallized using polyethylene glycol 3350 as a precipitant. In both complexes His280 of HsAtg4B was mutated to alanine. The crystals belong to the same space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 47.5, b = 91.8, c = 102.6 Å for complex I and a = 46.9, b = 90.9, c = 102.5 Å for complex II. Diffraction data were collected to a resolution of 1.9 Å from both crystals.

  7. Bacterial expression and preliminary crystallographic studies of a 149-residue fragment of human Caprin-1

    PubMed Central

    Wu, Yuhong; Zhu, Jiang; Huang, Xiaolan; Du, Zhihua

    2015-01-01

    Caprin-1 is an RNA-binding protein which plays critical roles in several important biological processes, including cellular proliferation, the interferon-mediated antiviral innate immune response, the maintenance of synaptic plasticity and the formation of RNA stress granules. Caprin-1 has been implicated in the pathogenesis of several human diseases, including osteo­sarcoma, breast cancer, viral infections, hearing loss and neurodegenerative disorders. Despite the emerging biological and physiopathological significance of Caprin-1, no structural information is available for this protein. Moreover, Caprin-1 does not have sequence similarity to any other protein with a known structure. It is therefore expected that structural studies will play a particularly crucial role in revealing the functional mechanisms of Caprin-1. Here, a protein fragment of human Caprin-1 consisting of residues 112–260 was expressed, purified and crystallized. Native and Se-SAD data sets were collected to resolutions to 2.05 and 2.65 Å, respectively, in different space groups. PMID:25760709

  8. Osteonic organization of limb bones in mammals, including humans, and birds: a preliminary study.

    PubMed

    Castrogiovanni, Paola; Imbesi, Rosa; Fisichella, Marco; Mazzone, Venera

    2011-01-01

    As it is well known, bone tissue is characterized by a calcified extracellular matrix which makes this tissue suitable to support the body and protect the inner organs. Lamellar bone tissue is organized in lamellae, 3-7 microm in thickness, and arranged concentrically around vascular channels: the basic structure in this type of organization is called Haversian system or osteon and the diameter of osteons depends on the number of lamellae. Shape and regional density of osteons are related to the bone segment and the specific functional requirements to meet. Aim of this study is to correlate the compact bone tissue microstructure in various classes of mammals, including humans, and birds in order to find an adequate identification key. The results of our study show that in bone tissue samples from various classes of mammals, including humans, and birds the osteonic structure shows peculiar features, often depending on the rate of bone remodelling, different in different animal species. We conclude that a careful microscopic analysis of bone tissue and the characterization of distinctive osteonic features could give a major contribution to forensic medicine to obtain a more reliable recognition of bone findings.

  9. Caffeine as an indicator of human fecal contamination in the Sinos River: a preliminary study.

    PubMed

    Linden, R; Antunes, M V; Heinzelmann, L S; Fleck, J D; Staggemeier, R; Fabres, R B; Vecchia, A D; Nascimento, C A; Spilki, F R

    2015-05-01

    The preservation of hydric resources is directly related to fecal contamination monitoring, in order to allow the development of strategies for the management of polluting sources. In the present study, twenty-five water samples from six water public supply collection sites were used for the evaluation of the presence of caffeine, total and fecal coliforms. Caffeine was detected in all samples, with concentrations ranging from 0.15 ng mL-1 to 16.72 ng mL-1. Total coliforms were detected in all samples, with concentrations in the range of 52 NMP/100 mL to higher than 24196 NMP/100 mL, whether the concentration range for fecal coliforms was in the range of below 1 NMP/100 mL to 7800 NMP/100 mL. No significant correlation was found between total coliforms and caffeine concentrations (rs = 0.35, p = 0.09). However, a moderate correlation between fecal coliforms and caffeine concentrations was found (rs = 0.412, p <0.05), probably indicating a human source for these bacteria. Caffeine determination in water may be a useful strategy to evaluate water contamination by human fecal waste.

  10. Purification, crystallization and preliminary X-ray characterization of the human GTP fucose pyrophosphorylase

    SciTech Connect

    Quirk, Stephen; Seley-Radtke, Katherine L.

    2006-04-01

    The human GTP fucose pyrophosphohydrolase protein has been crystallized via the hanging-drop technique over a reservoir of polyethylene glycol (MW 8000) and ethylene glycol. The orthorhombic crystals diffract to 2.8 Å resolution. The human nucleotide-sugar metabolizing enzyme GTP fucose pyrophosphorylase (GFPP) has been purified to homogeneity by an affinity chromatographic procedure that utilizes a novel nucleoside analog. This new purification regime results in a protein preparation that produces significantly better crystals than traditional purification methods. The purified 66.6 kDa monomeric protein has been crystallized via hanging-drop vapor diffusion at 293 K. Crystals of the native enzyme diffract to 2.8 Å and belong to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}. There is a single GFPP monomer in the asymmetric unit, giving a Matthews coefficient of 2.38 Å{sup 3} Da{sup −1} and a solvent content of 48.2%. A complete native data set has been collected as a first step in determining the three-dimensional structure of this enzyme.

  11. What Happened, and Why: Toward an Understanding of Human Error Based on Automated Analyses of Incident Reports. Volume 2

    NASA Technical Reports Server (NTRS)

    Ferryman, Thomas A.; Posse, Christian; Rosenthal, Loren J.; Srivastava, Ashok N.; Statler, Irving C.

    2006-01-01

    The objective of the Aviation System Monitoring and Modeling project of NASA's Aviation Safety and Security Program was to develop technologies to enable proactive management of safety risk, which entails identifying the precursor events and conditions that foreshadow most accidents. Information about what happened can be extracted from quantitative data sources, but the experiential account of the incident reporter is the best available source of information about why an incident happened. In Volume I, the concept of the Scenario was introduced as a pragmatic guide for identifying similarities of what happened based on the objective parameters that define the Context and the Outcome of a Scenario. In this Volume II, that study continues into the analyses of the free narratives to gain understanding as to why the incident occurred from the reporter s perspective. While this is just the first experiment, the results of our approach are encouraging and indicate that it will be possible to design an automated analysis process guided by the structure of the Scenario that can achieve the level of consistency and reliability of human analysis of narrative reports.

  12. Microarray Analyses Reveal Marked Differences in Growth Factor and Receptor Expression Between 8-Cell Human Embryos and Pluripotent Stem Cells.

    PubMed

    Vlismas, Antonis; Bletsa, Ritsa; Mavrogianni, Despina; Mamali, Georgina; Pergamali, Maria; Dinopoulou, Vasiliki; Partsinevelos, George; Drakakis, Peter; Loutradis, Dimitris; Kiessling, Ann A

    2016-01-15

    Previous microarray analyses of RNAs from 8-cell (8C) human embryos revealed a lack of cell cycle checkpoints and overexpression of core circadian oscillators and cell cycle drivers relative to pluripotent human stem cells [human embryonic stem cells/induced pluripotent stem (hES/iPS)] and fibroblasts, suggesting growth factor independence during early cleavage stages. To explore this possibility, we queried our combined microarray database for expression of 487 growth factors and receptors. Fifty-one gene elements were overdetected on the 8C arrays relative to hES/iPS cells, including 14 detected at least 80-fold higher, which annotated to multiple pathways: six cytokine family (CSF1R, IL2RG, IL3RA, IL4, IL17B, IL23R), four transforming growth factor beta (TGFB) family (BMP6, BMP15, GDF9, ENG), one fibroblast growth factor (FGF) family [FGF14(FH4)], one epidermal growth factor member (GAB1), plus CD36, and CLEC10A. 8C-specific gene elements were enriched (73%) for reported circadian-controlled genes in mouse tissues. High-level detection of CSF1R, ENG, IL23R, and IL3RA specifically on the 8C arrays suggests the embryo plays an active role in blocking immune rejection and is poised for trophectoderm development; robust detection of NRG1, GAB1, -2, GRB7, and FGF14(FHF4) indicates novel roles in early development in addition to their known roles in later development. Forty-four gene elements were underdetected on the 8C arrays, including 11 at least 80-fold under the pluripotent cells: two cytokines (IFITM1, TNFRSF8), five TGFBs (BMP7, LEFTY1, LEFTY2, TDGF1, TDGF3), two FGFs (FGF2, FGF receptor 1), plus ING5, and WNT6. The microarray detection patterns suggest that hES/iPS cells exhibit suppressed circadian competence, underexpression of early differentiation markers, and more robust expression of generic pluripotency genes, in keeping with an artificial state of continual uncommitted cell division. In contrast, gene expression patterns of the 8C embryo suggest that

  13. Microarray Analyses Reveal Marked Differences in Growth Factor and Receptor Expression Between 8-Cell Human Embryos and Pluripotent Stem Cells

    PubMed Central

    Vlismas, Antonis; Bletsa, Ritsa; Mavrogianni, Despina; Mamali, Georgina; Pergamali, Maria; Dinopoulou, Vasiliki; Partsinevelos, George; Drakakis, Peter; Loutradis, Dimitris

    2016-01-01

    Previous microarray analyses of RNAs from 8-cell (8C) human embryos revealed a lack of cell cycle checkpoints and overexpression of core circadian oscillators and cell cycle drivers relative to pluripotent human stem cells [human embryonic stem cells/induced pluripotent stem (hES/iPS)] and fibroblasts, suggesting growth factor independence during early cleavage stages. To explore this possibility, we queried our combined microarray database for expression of 487 growth factors and receptors. Fifty-one gene elements were overdetected on the 8C arrays relative to hES/iPS cells, including 14 detected at least 80-fold higher, which annotated to multiple pathways: six cytokine family (CSF1R, IL2RG, IL3RA, IL4, IL17B, IL23R), four transforming growth factor beta (TGFB) family (BMP6, BMP15, GDF9, ENG), one fibroblast growth factor (FGF) family [FGF14(FH4)], one epidermal growth factor member (GAB1), plus CD36, and CLEC10A. 8C-specific gene elements were enriched (73%) for reported circadian-controlled genes in mouse tissues. High-level detection of CSF1R, ENG, IL23R, and IL3RA specifically on the 8C arrays suggests the embryo plays an active role in blocking immune rejection and is poised for trophectoderm development; robust detection of NRG1, GAB1, -2, GRB7, and FGF14(FHF4) indicates novel roles in early development in addition to their known roles in later development. Forty-four gene elements were underdetected on the 8C arrays, including 11 at least 80-fold under the pluripotent cells: two cytokines (IFITM1, TNFRSF8), five TGFBs (BMP7, LEFTY1, LEFTY2, TDGF1, TDGF3), two FGFs (FGF2, FGF receptor 1), plus ING5, and WNT6. The microarray detection patterns suggest that hES/iPS cells exhibit suppressed circadian competence, underexpression of early differentiation markers, and more robust expression of generic pluripotency genes, in keeping with an artificial state of continual uncommitted cell division. In contrast, gene expression patterns of the 8C embryo suggest that

  14. Purification, crystallization and preliminary X-ray diffraction of human S100A15

    SciTech Connect

    Boeshans, Karen M.; Wolf, Ronald; Voscopoulos, Christopher; Gillette, William; Esposito, Dominic; Mueser, Timothy C.; Yuspa, Stuart H.; Ahvazi, Bijan

    2006-05-01

    S100 proteins are differentially expressed during epithelial cell maturation, tumorigenesis and inflammation. The novel human S100A15 protein has been cloned, expressed, purified and crystallized in two crystal forms, a triclinic and a monoclinic form, which diffract to 1.7 and 2.0 Å, respectively. Human S100A15 is a novel member of the S100 family of EF-hand calcium-binding proteins and was recently identified in psoriasis, where it is significantly upregulated in lesional skin. The protein is implicated as an effector in calcium-mediated signal transduction pathways. Although its biological function is unclear, the association of the 11.2 kDa S100A15 with psoriasis suggests that it contributes to the pathogenesis of the disease and could provide a molecular target for therapy. To provide insight into the function of S100A15, the protein was crystallized to visualize its structure and to further the understanding of how the many similar calcium-binding mediator proteins in the cell distinguish their cognate target molecules. The S100A15 protein has been cloned, expressed and purified to homogeneity and produced two crystal forms. Crystals of form I are triclinic, with unit-cell parameters a = 33.5, b = 44.3, c = 44.8 Å, α = 71.2, β = 68.1, γ = 67.8° and an estimated two molecules in the asymmetric unit, and diffract to 1.7 Å resolution. Crystals of form II are monoclinic, with unit-cell parameters a = 82.1, b = 33.6, c = 52.2 Å, β = 128.2° and an estimated one molecule in the asymmetric unit, and diffract to 2.0 Å resolution. This structural analysis of the human S100A15 will further aid in the phylogenic comparison between the other members of the S100 protein family, especially the highly homologous paralog S100A7.

  15. Crystallization and preliminary neutron diffraction experiment of human farnesyl pyrophosphate synthase complexed with risedronate

    PubMed Central

    Yokoyama, Takeshi; Ostermann, Andreas; Mizuguchi, Mineyuki; Niimura, Nobuo; Schrader, Tobias E.; Tanaka, Ichiro

    2014-01-01

    Nitrogen-containing bisphosphonates (N-BPs), such as risedronate and zoledronate, are currently used as a clinical drug for bone-resorption diseases and are potent inhibitors of farnesyl pyrophosphate synthase (FPPS). X-ray crystallographic analyses of FPPS with N-BPs have revealed that N-BPs bind to FPPS with three magnesium ions and several water molecules. To understand the structural characteristics of N-BPs bound to FPPS, including H atoms and hydration by water, neutron diffraction studies were initiated using BIODIFF at the Heinz Maier-Leibnitz Zentrum (MLZ). FPPS–risedronate complex crystals of approximate dimensions 2.8 × 2.5 × 1.5 mm (∼3.5 mm3) were obtained by repeated macro-seeding. Monochromatic neutron diffraction data were collected to 2.4 Å resolution with 98.4% overall completeness. Here, the first successful neutron data collection from FPPS in complex with N-BPs is reported. PMID:24699741

  16. Crystallization and preliminary neutron diffraction experiment of human farnesyl pyrophosphate synthase complexed with risedronate.

    PubMed

    Yokoyama, Takeshi; Ostermann, Andreas; Mizuguchi, Mineyuki; Niimura, Nobuo; Schrader, Tobias E; Tanaka, Ichiro

    2014-04-01

    Nitrogen-containing bisphosphonates (N-BPs), such as risedronate and zoledronate, are currently used as a clinical drug for bone-resorption diseases and are potent inhibitors of farnesyl pyrophosphate synthase (FPPS). X-ray crystallographic analyses of FPPS with N-BPs have revealed that N-BPs bind to FPPS with three magnesium ions and several water molecules. To understand the structural characteristics of N-BPs bound to FPPS, including H atoms and hydration by water, neutron diffraction studies were initiated using BIODIFF at the Heinz Maier-Leibnitz Zentrum (MLZ). FPPS-risedronate complex crystals of approximate dimensions 2.8 × 2.5 × 1.5 mm (∼3.5 mm(3)) were obtained by repeated macro-seeding. Monochromatic neutron diffraction data were collected to 2.4 Å resolution with 98.4% overall completeness. Here, the first successful neutron data collection from FPPS in complex with N-BPs is reported.

  17. Microarray data and pathway analyses for primary human activated hepatic stellate cells compared to HepG2 human hepatoma cells.

    PubMed

    Hetherington, Alexandra M; Sawyez, Cynthia G; Borradaile, Nica M

    2017-02-01

    As nonalcoholic fatty liver disease progresses to end-stage diseases, including fibrosis, cirrhosis and hepatocellular carcinoma, fibrotic activated hepatic stellate cells and cancerous epithelial cells can become abundant, changing the cellular composition of this organ. Despite potentially residing within the same diseased tissue, direct comparisons of global gene expression between activated hepatic stellate cells and hepatocellular carcinoma cells are lacking. Here we provide data collected using Affymetrix GeneChip microarrays to identify differential gene expression in cultured primary human activated hepatic stellate cells compared to HepG2 human hepatoma cells. The dataset includes many genes involved in intermediary metabolism which were investigated in greater depth in our associated article (A.M. Hetherington, C.G. Sawyez, E. Zilberman, A.M. Stoianov, D.L. Robson, J.M. Hughes-Large, et al., 2016) [1]. Pathway analyses of known protein coding genes down-regulated or up-regulated by greater than 2.0-fold are also provided.

  18. Crystallization and preliminary X-ray crystallographic analysis of human myotubularin-related protein 1

    PubMed Central

    Bong, Seoung Min; Yang, Seung Won; Choi, Ji-Woong; Kim, Seung Jun; Lee, Byung Il

    2015-01-01

    Myotubularin-related protein 1 is a phosphatase that dephosphorylates phospholipids such as phosphatidylinositol 3-phosphate or phosphatidylinositol 3,5-bisphosphate. In this study, human MTMR1 was overexpressed in Escherichia coli, purified and crystallized at 277 K using polyethylene glycol 20 000 as a precipitant. Diffraction data were collected to 2.0 Å resolution using synchrotron radiation. The crystals belonged to space group P1, with unit-cell parameters a = 67.219, b = 96.587, c = 97.581 Å, α = 87.597, β = 86.072, γ = 77.327°. Assuming the presence of four molecules in the asymmetric unit, the calculated Matthews coefficient value was 2.61 Å3 Da−1 and the corresponding solvent content was 52.9%. PMID:25760698

  19. Bodyworlds and the ethics of using human remains: a preliminary discussion.

    PubMed

    Barilan, Y Michael

    2006-09-01

    Accepting the claim that the living have some moral duties with regard to dead bodies, this paper explores those duties and how they bear on the popular travelling exhibition Bodyworlds. I argue that the concept of informed consent presupposes substantial duties to the dead, namely duties that reckon with the meaning of the act in question. An attitude of respect and not regarding human remains as mere raw material are non-alienable substantial duties. I found the ethos of Bodyworlds premature but full of promises such as public attitudes to organ donations. At the practical level I conclude that Bodyworlds should use only willed donations or unclaimed bodies for which dignified funerals are not available. In the case of live donations, Bodyworlds has a duty to participate in the medical care of needy donors. However, secrecy with regard to the source of cadavers seems to be the most troublesome aspect of Bodyworlds.

  20. Rimonabant effects on anxiety induced by simulated public speaking in healthy humans: a preliminary report

    PubMed Central

    Bergamaschi, Mateus M.; Queiroz, Regina H. C.; Chagas, Marcos H. N.; Linares, Ila M. P.; Arrais, Kátia C.; de Oliveira, Danielle C. G.; Queiroz, Maria E.; Nardi, Antonio E.; Huestis, Marilyn A.; Hallak, Jaime E. C.; Zuardi, Antonio W.; Moreira, Fabrício A.; Crippa, José A. S.

    2015-01-01

    Objective We investigated the hypothesis that rimonabant, a cannabinoid antagonist/inverse agonist, would increase anxiety in healthy subjects during a simulation of the public speaking test. Methods Participants were randomly allocated to receive oral placebo or 90 mg rimonabant in a double-blind design. Subjective effects were measured by Visual Analogue Mood Scale. Physiological parameters, namely arterial blood pressure and heart rate, also were monitored. Results Twelve participants received oral placebo and 12 received 90 mg rimonabant. Rimonabant increased self-reported anxiety levels during the anticipatory speech and performance phase compared with placebo. Interestingly, rimonabant did not modulate anxiety prestress and was not associated with sedation, cognitive impairment, discomfort, or blood pressure changes. Conclusions Cannabinoid-1 antagonism magnifies the responses to an anxiogenic stimulus without interfering with the prestress phase. These data suggest that the endocannabinoid system may work on-demand to counteract the consequences of anxiogenic stimuli in healthy humans. PMID:24424711

  1. Crystallization and Preliminary X-Ray Crystallographic Analysis of Human Plasma Platelet Activating Factor Acetylhydrolase

    SciTech Connect

    Samanta, U.; Wilder, C; Bahnson, B

    2009-01-01

    The plasma form of the human enzyme platelet activating factor acetylhydrolase (PAF-AH) has been crystallized, and X-ray diffraction data were collected at a synchrotron source to a resolution of 1.47 {angstrom}. The crystals belong to space group C2, with unit cell parameters of a = 116.18, b = 83.06, c = 96.71 {angstrom}, and {beta} = 115.09 and two molecules in the asymmetric unit. PAF-AH functions as a general anti-inflammatory scavenger by reducing the levels of the signaling molecule PAF. Additionally, the LDL bound enzyme has been linked to atherosclerosis due to its hydrolytic activities of pro-inflammatory agents, such as sn-2 oxidatively fragmented phospholipids.

  2. Preliminary System Analysis of In Situ Resource Utilization for Mars Human Exploration

    NASA Technical Reports Server (NTRS)

    Rapp, Donald; Andringa, Jason; Easter, Robert; Smith, Jeffrey H .; Wilson, Thomas; Clark, D. Larry; Payne, Kevin

    2005-01-01

    We carried out a system analysis of processes for utilization of Mars resources to support human exploration of Mars by production of propellants from indigenous resources. Seven ISRU processes were analyzed to determine mass. power and propellant storage volume requirements. The major elements of each process include C02 acquisition, chemical conversion, and storage of propellants. Based on a figure of merit (the ratio of the mass of propellants that must be brought from Earth in a non-ISRU mission to the mass of the ISRU system. tanks and feedstocks that must be brought from Earth for a ISRU mission) the most attractive process (by far); is one where indigenous Mars water is accessible and this is processed via Sabatier/Electrolysis to methane and oxygen. These processes are technically relatively mature. Other processes with positive leverage involve reverse water gas shift and solid oxide electrolysis.

  3. National Human Research Ethics: A Preliminary Comparative Case Study of Germany, Great Britain, Romania, and Sweden

    PubMed Central

    Gallagher, Bernard; Berman, Anne H.; Bieganski, Justyna; Jones, Adele D.; Foca, Liliana; Raikes, Ben; Schiratzki, Johanna; Urban, Mirjam; Ullman, Sara

    2016-01-01

    Although international research is increasing in volume and importance, there remains a dearth of knowledge on similarities and differences in “national human research ethics” (NHREs), that is, national ethical guidelines (NEGs), Institutional Review Boards (IRBs), and research stakeholder’ ethical attitudes and behaviors (EABs). We begin to address this situation by reporting upon our experiences in conducting a multinational study into the mental health of children who had a parent/carer in prison. The study was conducted in 4 countries: Germany, Great Britain, Romania, and Sweden. Data on NHREs were gathered via a questionnaire survey, two ethics-related seminars, and ongoing contact between members of the research consortium. There was correspondence but even more so divergence between countries in the availability of NEGs and IRBs and in researcher’ EABs. Differences in NHREs have implications particularly in terms of harmonization but also for ethical philosophy and practice and for research integrity. PMID:27746664

  4. Preliminary Toxicological Analysis of the Effect of Coal Slurry Impoundment Water on Human Liver Cells

    USGS Publications Warehouse

    Bunnell, Joseph E.

    2008-01-01

    Coal is usually 'washed' with water and a variety of chemicals to reduce its content of sulfur and mineral matter. The 'washings' or 'coal slurry' derived from this process is a viscous black liquid containing fine particles of coal, mineral matter, and other dissolved and particulate substances. Coal slurry may be stored in impoundments or in abandoned underground mines. Human health and environmental effects potentially resulting from leakage of chemical substances from coal slurry into drinking water supplies or aquatic ecosystems have not been systematically examined. Impoundments are semipermeable, presenting the possibility that inorganic and organic substances, some of which may be toxic, may contaminate ground or surface water. The Agency for Toxic Substances and Disease Registry, part of the Centers for Disease Control and Prevention, has concluded that well water in Mingo County, West Virginia, constitutes a public health hazard.

  5. Human imprint on archaeological anthroposols: first assessment of combined micromophological, pedological and lipid biomarkers analyses of organic matter

    NASA Astrophysics Data System (ADS)

    Cammas, Cécilia; Thuy Nguyen Tu, Thanh; Plessis, Marion; Clotuche, Raphaël; Derenne, Sylvie

    2013-04-01

    Archaeological anthroposol matrix contains significant amounts of fine organic matter (OM), which can give archaeological information. Geoarchaeological studies of OM aim to reveal its origin in order to reconstruct past human activities. Such studies are complex because the nature and the abundance of OM is the result of human activities together with natural processes. Also, MO evolves over time, a process that is not well understood. Combination of complementary approaches may give further insights into human imprint on archaeological anthroposols. For example, micromorphology gives data on in situ activities and pedological processes with the result that components of animal and vegetal origin can be identified but not some amorphous / fibrous material and very fine residues (< ~10 µm). On the other hand, pedological and geochemical analyses of bulk samples are often disconnected from contextual studies. Our work aims to (i) identify morphological and geochemical markers of human activity at different scales, (ii) compare results of different analytical methods to better understand the relation between matrix components and features, chemical properties, and geochemical markers, and (iii) infer relations between pedo-sedimentary history and OM preservation. Two tanning pits in urban craft areas were selected for sampling, as they are likely to contain large amounts of organic matter of vegetal and animal origin. The pit of Saint-Denis (SDN, 10 km at the north of Paris, calcareous alluvium, 13th cAD) was a reference tanning pit. The pit of Famars (FAM, near the Belgian border, luvisols, Roman period) was hypothesized to be a part of the tanning process. To assess preservation of organic components and molecules in relation with pedo-sedimentary context and their potential as biomarkers of human activities, methodology combined micromorphology, pedological analysis (C, N, LOI, P total, organic and inorganic phosphorus) and lipid analysis by GC/MS, lipids having

  6. A feasibility study of soft embalmed human breast tissue for preclinical trials of HIFU- preliminary results

    NASA Astrophysics Data System (ADS)

    Joy, Joyce; Yang, Yang; Purdie, Colin; Eisma, Roos; Melzer, Andreas; Cochran, Sandy; Vinnicombe, Sarah

    2017-03-01

    Breast cancer is the commonest cancer in women in the UK, accounting for 30% of all new cancers in women, with an estimated 49,500 new cases in 20101. With the widespread negative publicity around over-diagnosis and over-treatment of low risk breast cancers, interest in the application of non-invasive treatments such as magnetic resonance imaging (MRI) guided high intensity focused ultrasound (HIFU) has increased. Development has begun of novel US transducers and platforms specifically designed for use with breast lesions, so as to improve the range of breast lesions that can be safely treated. However, before such transducers can be evaluated in patients in clinical trials, there is a need to establish their efficacy. A particular issue is the accuracy of temperature monitoring of FUS with MRI in the breast, since the presence of large amounts of surrounding fat can hinder temperature measurement. An appropriate anatomical model that imposes similar physical constraints to the breast and that responds to FUS in the same way would be extremely advantageous. The aim of this feasibility study is to explore the use of Thiel embalmed cadaveric tissue for these purposes. We report here the early results of laboratory-based experiments sonicating dissected breast samples from a Thiel embalmed soft human cadaver with high body mass index (BMI). A specially developed MRI compatible chamber and sample holder was developed to secure the sample and ensure reproducible sonications at the transducer focus. The efficacy of sonication was first studied with chicken breast and porcine tissue. The experiments were then repeated with the dissected fatty breast tissue samples from the soft-embalmed human cadavers. The sonicated Thiel breast tissue was examined histopathologically, which confirmed the absence of any discrete lesion. To investigate further, fresh chicken breast tissue was embalmed and the embalmed tissue was sonicated with the same parameters. The results confirmed the

  7. Purification, crystallization and preliminary X-ray diffraction analysis of the human mismatch repair protein MutS[beta

    SciTech Connect

    Tseng, Quincy; Orans, Jillian; Hast, Michael A.; Iyer, Ravi R.; Changela, Anita; Modrich, Paul L.; Beese, Lorena S.

    2012-03-16

    MutS{beta} is a eukaryotic mismatch repair protein that preferentially targets extrahelical unpaired nucleotides and shares partial functional redundancy with MutS{alpha} (MSH2-MSH6). Although mismatch recognition by MutS{alpha} has been shown to involve a conserved Phe-X-Glu motif, little is known about the lesion-binding mechanism of MutS{beta}. Combined MSH3/MSH6 deficiency triggers a strong predisposition to cancer in mice and defects in msh2 and msh6 account for roughly half of hereditary nonpolyposis colorectal cancer mutations. These three MutS homologs are also believed to play a role in trinucleotide repeat instability, which is a hallmark of many neurodegenerative disorders. The baculovirus overexpression and purification of recombinant human MutS{beta} and three truncation mutants are presented here. Binding assays with heteroduplex DNA were carried out for biochemical characterization. Crystallization and preliminary X-ray diffraction analysis of the protein bound to a heteroduplex DNA substrate are also reported.

  8. Complex assembly, crystallization and preliminary X-ray crystallographic analysis of the human Rod–Zwilch–ZW10 (RZZ) complex

    SciTech Connect

    Altenfeld, Anika; Wohlgemuth, Sabine; Wehenkel, Annemarie; Musacchio, Andrea

    2015-03-20

    The 800 kDa complex of the human Rod, Zwilch and ZW10 proteins (the RZZ complex) was reconstituted in insect cells, purified, crystallized and subjected to preliminary X-ray diffraction analysis. The spindle-assembly checkpoint (SAC) monitors kinetochore–microtubule attachment during mitosis. In metazoans, the three-subunit Rod–Zwilch–ZW10 (RZZ) complex is a crucial SAC component that interacts with additional SAC-activating and SAC-silencing components, including the Mad1–Mad2 complex and cytoplasmic dynein. The RZZ complex contains two copies of each subunit and has a predicted molecular mass of ∼800 kDa. Given the low abundance of the RZZ complex in natural sources, its recombinant reconstitution was attempted by co-expression of its subunits in insect cells. The RZZ complex was purified to homogeneity and subjected to systematic crystallization attempts. Initial crystals containing the entire RZZ complex were obtained using the sitting-drop method and were subjected to optimization to improve the diffraction resolution limit. The crystals belonged to space group P3{sub 1} (No. 144) or P3{sub 2} (No. 145), with unit-cell parameters a = b = 215.45, c = 458.7 Å, α = β = 90.0, γ = 120.0°.

  9. Subject-specific estimation of central aortic blood pressure via system identification: preliminary in-human experimental study.

    PubMed

    Fazeli, Nima; Kim, Chang-Sei; Rashedi, Mohammad; Chappell, Alyssa; Wang, Shaohua; MacArthur, Roderick; McMurtry, M Sean; Finegan, Barry; Hahn, Jin-Oh

    2014-10-01

    This paper demonstrates preliminary in-human validity of a novel subject-specific approach to estimation of central aortic blood pressure (CABP) from peripheral circulatory waveforms. In this "Individualized Transfer Function" (ITF) approach, CABP is estimated in two steps. First, the circulatory dynamics of the cardiovascular system are determined via model-based system identification, in which an arterial tree model is characterized based on the circulatory waveform signals measured at the body's extremity locations. Second, CABP waveform is estimated by de-convolving peripheral circulatory waveforms from the arterial tree model. The validity of the ITF approach was demonstrated using experimental data collected from 13 cardiac surgery patients. Compared with the invasive peripheral blood pressure (BP) measurements, the ITF approach yielded significant reduction in errors associated with the estimation of CABP, including 1.9-2.6 mmHg (34-42 %) reduction in BP waveform errors (p < 0.05) as well as 5.8-9.1 mmHg (67-76 %) and 6.0-9.7 mmHg (78-85 %) reductions in systolic and pulse pressure (SP and PP) errors (p < 0.05). It also showed modest but significant improvement over the generalized transfer function approach, including 0.1 mmHg (2.6 %) reduction in BP waveform errors as well as 0.7 (20 %) and 5.0 mmHg (75 %) reductions in SP and PP errors (p < 0.05).

  10. Human adaptation to isolated and confined environments: Preliminary findings of a seven month Antarctic winter-over human factors study

    NASA Technical Reports Server (NTRS)

    Evans, Gary W.; Stokols, Daniel; Carrere, Sybil

    1988-01-01

    This field study was conducted during the last decade of an austral winter-over at Palmer Station in the Antarctic. The purpose of the study was to understand temporal patterns in physiological arousal and psychological mood over the course of the mission. The investigators were principally interested in how people adapted over time to chronic and acute stressors, and how people use and modify their built environment. Physiological and psychological data were collected several times a week, and information on behavior and the use of physical facilities was collected monthly. Physiological and psychological data were compared with social changes in the setting toward the development of a sequential model of human-environment transactional relationships. Based on the study results, guidelines for design of future isolated and confined environments (ICEs) included: plan space for items which make people feel at home, provide materials to allow people to personalize their environment, allow for flexible environments, provide areas for visual and auditory privacy, equip areas for socializing and remove them from private areas, and provide facilities for exercise and for projects involving physical activity. The study offers guidelines about patterns of adaption that could be expected in an ICE, discusses how these settings can be programmed to facilitate successful adjustment, and provides information about how to design future ICE habitats to maximize a healthy living environment.

  11. The effect of alpha-lipoic acid on temporary threshold shift in humans: a preliminary study.

    PubMed

    Quaranta, N; Dicorato, A; Matera, V; D'Elia, A; Quaranta, A

    2012-12-01

    Noise-induced hearing loss (NHIL) is a significant source of hearing loss in industrialized countries. Recent research on the cellular bases of NIHL has led to new avenues for protection through prophylactic drugs. Although in experimental animal models several compounds have shown a protective effect in NIHL, limited data are available in humans. Many authors are focusing their attention on the role of antioxidant on hearing protection. Alpha-lipoic acid (ALA), an essential cofactor in mitochondrial enzymes, is a novel biological antioxidant and a potent free radical scavenger and, in animal models, it has been shown to protect from age-induced and cisplatin-induced hearing loss. The aim of our study was to evaluate the effect of alpha-lipoic acid on temporary threshold shift measured 2 minutes after the end of exposure (TTS(2)) induced by a 3 kHz tone in young normally hearing subjects. Thirty young normal hearing volunteers served as control subjects. Individuals were randomly assigned to three groups. Group A (10 subjects) subjects were exposed to a 90 dB HL 3 kHz pure tone for 10 min. Group B (10 subjects) subjects were exposed to a 90 dB HL 3 kHz pure tone one hour after oral ingestion of 600 mg of ALA. Group C (10 subjects) were exposed to a 90 dB HL 3 kHz pure tone after 10 days of oral ingestion of 600 mg of ALA. Statistical analysis showed that prior to the exposure the hearing thresholds did not differ significantly among the three groups. TTS(2) of group C was significantly lower that TTS2 of Groups A and B at 6 kHz (p 0.03), and TEOAEs amplitude change after noise exposure was lower for group C compared to Groups A (p = 0.089) and B (p = 0.03). ALA is a powerful lipophilic antioxidant and free radical scavenger currently used in clinical practice. A single dose of 600 mg of dose ALA did not induce any protection on the TTS(2) induced by a 90 dB HL 3 kHz tone, while 10 days of therapeutic dosage assumption of ALA was associated with significant

  12. Preliminary results of Physiological plant growth modelling for human life support in space

    NASA Astrophysics Data System (ADS)

    Sasidharan L, Swathy; Dussap, Claude-Gilles; Hezard, Pauline

    2012-07-01

    Human life support is fundamental and crucial in any kind of space explorations. MELiSSA project of European Space Agency aims at developing a closed, artificial ecological life support system involving human, plants and micro organisms. Consuming carbon dioxide and water from the life support system, plants grow in one of the chambers and convert it into food and oxygen along with potable water. The environmental conditions, nutrient availability and its consumption of plants should be studied and necessarily modeled to predict the amount of food, oxygen and water with respect to the environmental changes and limitations. The reliability of a completely closed system mainly depends on the control laws and strategies used. An efficient control can occur, only if the system to control is itself well known, described and ideally if the responses of the system to environmental changes are predictable. In this aspect, the general structure of plant growth model has been designed together with physiological modelling.The physiological model consists of metabolic models of leaves, stem and roots, of which concern specific metabolisms of the associated plant parts. On the basis of the carbon source transport (eg. sucrose) through stem, the metabolic models (leaf and root) can be interconnected to each other and finally coupled to obtain the entire plant model. For the first step, leaf metabolic model network was built using stoichiometric, mass and energy balanced metabolic equations under steady state approach considering all necessary plant pathways for growth and maintenance of leaves. As the experimental data for lettuce plants grown in closed and controlled environmental chambers were available, the leaf metabolic model has been established for lettuce leaves. The constructed metabolic network is analyzed using known stoichiometric metabolic technique called metabolic flux analysis (MFA). Though, the leaf metabolic model alone is not sufficient to achieve the

  13. Crystallization and preliminary crystallographic studies of human septin 1 with site-directed mutations

    SciTech Connect

    Hu, Hao; Yu, Wen-bo; Li, Shu-xing; Ding, Xiang-ming; Yu, Long; Bi, Ru-Chang

    2006-02-01

    The homogeneity of septin 1 has been improved by site-directed mutation of serine residues and only a small alteration in the secondary structure is observed to arise from the mutations. Crystals of the septin 1 mutant were grown and diffraction data were collected to 2.5 Å resolution. Septin 1 is a member of an evolutionarily conserved family of GTP-binding and filament-forming proteins named septins, which function in diverse processes including cytokinasis, vesicle trafficking, apoptosis, remodelling of the cytoskeleton, infection, neurodegeneration and neoplasia. Human septin 1 has been expressed and purified, but suffers from severe aggregation. Studies have shown that septin 1 with site-directed mutations of five serine residues (Ser19, Ser206, Ser307, Ser312 and Ser315) has a much lower degree of aggregation and better structural homogeneity and that the mutations cause only slight perturbations in the secondary structure of septin 1. This septin 1 mutant was crystallized and diffraction data were collected to 2.5 Å resolution. The space group is P422, with unit-cell parameters a = b = 106.028, c = 137.852 Å.

  14. Daily feeding rhythm in proboscis monkeys: a preliminary comparison with other non-human primates.

    PubMed

    Matsuda, Ikki; Akiyama, Yoshihiro; Tuuga, Augustine; Bernard, Henry; Clauss, Marcus

    2014-04-01

    In non-human primates, the daily feeding rhythm, i.e., temporal fluctuation in feeding activity across the day, has been described but has rarely received much analytical interpretation, though it may play a crucial part in understanding the adaptive significance of primate foraging strategies. This study is the first to describe the detailed daily feeding rhythm in proboscis monkeys (Nasalis larvatus) based on data collected from both riverbank and inland habitats. From May 2005 to May 2006, data on feeding behavior in a group of proboscis monkeys consisting of an alpha-male, six adult females and immatures was collected via continuous focal animal sampling technique in a forest along the Menanggul River, Sabah, Malaysia. In both the male and females, the highest peak of feeding activity was in the late afternoon at 15:00-17:00, i.e., shortly before sleeping. The differences in the feeding rhythm among the seasons appeared to reflect the time spent eating fruit and/or the availability of fruit; clearer feeding peaks were detected when the monkeys spent a relevant amount of time eating fruit, but no clear peak was detected when fruit eating was less frequent. The daily feeding rhythm was not strongly influenced by daily temperature fluctuations. When comparing the daily feeding rhythm of proboscis monkeys to that of other primates, one of the most common temporal patterns detected across primates was a feeding peak in the late afternoon, although it was impossible to demonstrate this statistically because of methodological differences among studies.

  15. A preliminary insight of correlation between human fecal microbial diversity and blood lipid profile.

    PubMed

    Madeeha, Ilyas Rana; Ikram, Aamer; Imran, Muhammad

    2016-11-01

    The study aimed to evaluate the effect of human gut-derived lactic acid bacteria and yeast on cholesterol levels. Fecal samples from five healthy volunteers were examined for the level and diversity of dominant microbiota. Pichia kudriavzevii (QAUPK01, QAUPK02, QAUPK03, QAUPK04 and QAUPK05) and Candida tropicalis (QAUCT06) were identified by phenotypic methods and DNA sequencing and tested for in vitro cholesterol assimilation ability. Significant correlations (p < 0.05) between fecal microbial diversity, volunteers' age, body mass index (BMI) and serum lipid profile were established. From biochemical tests, eight strains of lactic acid bacteria (M1.1, M1.2, M2.1, M3.1, M3.2, M4.1, M5.1 and M5.2) were identified but no bsh activity was found in them. However, all yeast strains were able to assimilate cholesterol and maximum assimilation ability was shown by QAUPK03 (83.6%) and QAUPK05 (85.2%) after 72 h of growth at 37 °C.

  16. Crystallization and preliminary crystallographic study of a trypsin-resistant catalytic domain of human calcineurin

    PubMed Central

    Jin, Lei; Roehrl, Michael H. A.; Xiao, Li; He, Xiuyun; Li, Haibin; Ge, Linhu; Shi, Bingyi

    2012-01-01

    Calcineurin, a Ca2+/calmodulin-dependent serine/threonine protein phosphatase, plays a key role in a number of cellular pathways, including T-cell activation, and is an important molecular target of the immunosuppressive drugs cyclosporin A and FK506. To understand the structural basis underlying the activation of calcineurin by calmodulin, X-ray crystallography was employed to solve the three-dimensional structure of the free calcineurin catalytic domain (residues 20–347 of the A subunit). To accomplish this, a bacterially expressed glutathione S-­transferase (GST) fusion protein of the human calcineurin catalytic domain was first purified by GST-affinity chromatography. After limited digestion by trypsin, the catalytic domain (Cncat) was purified using anion-exchange and size-exclusion chromatography. Crystallization of Cncat was achieved by the hanging-drop vapour-diffusion method at pH 6.5 using PEG 6000 as precipitant. The diffraction results showed that the Cncat crystal belonged to the orthorhombic space group P21212, with unit-cell parameters a = 161.6, b = 87.4, c = 112.0 Å. There are four Cncat molecules in the asymmetric unit, with 49.5% solvent content. An X-ray diffraction data set was collected to 2.87 Å resolution and a clear molecular-replacement solution was obtained. The active site of Cncat is open to the solvent channels in the crystal packing. PMID:22691791

  17. Preliminary Human-in-the-Loop Assessment of Procedures for Very-Closely-Spaced Parallel Runways

    NASA Technical Reports Server (NTRS)

    Verma, Savita; Lozito, Sandra C.; Ballinger, Deborah S.; Trot, Greg; Hardy, Gordon H.; Panda, Ramesh C.; Lehmer, Ronald D.; Kozon, Thomas E.

    2010-01-01

    Demand in the future air transportation system concept is expected to double or triple by 2025 [1]. Increasing airport arrival rates will help meet the growing demand that could be met with additional runways but the expansion airports is met with environmental challenges for the surrounding communities when using current standards and procedures. Therefore, changes to airport operations can improve airport capacity without adding runways. Building additional runways between current ones, or moving them closer, is a potential solution to meeting the increasing demand, as addressed by the Terminal Area Capacity Enhancing Concept (TACEC). TACEC requires robust technologies and procedures that need to be tested such that operations are not compromised under instrument meteorological conditions. The reduction of runway spacing for independent simultaneous operations dramatically exacerbates the criticality of wake vortex incursion and the calculation of a safe and proper breakout maneuver. The study presented here developed guidelines for such operations by performing a real-time, human-in-the-loop simulation using precision navigation, autopilot-flown approaches, with the pilot monitoring aircraft spacing and the wake vortex safe zone during the approach.

  18. Prevalence of human T-cell lymphotropic virus type I infection in Singapore: a preliminary report.

    PubMed

    Zhao, L G; Yanagihara, R; Mora, C; Garruto, R M; Wong, T W; Gajdusek, D C

    1991-01-01

    Human T-cell lymphotropic virus type I (HTLV-I) infection is endemic in southwestern Japan, the Caribbean basin, Colombia, Africa and in several isolated populations in Papua New Guinea, the Solomon Islands and Vanuatu. To determine the seroprevalence of HTLV-I infection in Singapore, we tested sera from 115 hospitalized patients with acute nephritis, 50 patients with suspected leptospirosis, 34 patients with non-A, non-B hepatitis, and from 28 healthy volunteers for IgG antibodies against HTLV-I using an enzyme-linked immunosorbent assay. Antibodies were detected in sera from 6 of the 199 patients and from 3 of the 28 healthy volunteers, but these positives could not be confirmed by Western immunoblotting. Our data are consistent with other reports of low seroprevalence of HTLV-I infection despite extensive Japanese contact in Korea, Taiwan, the People's Republic of China and Micronesia. Further studies on a larger sample size, however, are necessary to confirm the absence of any focus of infection in the Singapore population.

  19. Human Lunar Mission Capabilities Using SSTO, ISRU and LOX-Augmented NTR Technologies: A Preliminary Assessment

    NASA Technical Reports Server (NTRS)

    Borowski, Stanley K.

    1995-01-01

    The feasibility of conducting human missions to the Moon is examined assuming the use of three 'high leverage' technologies: (1) a single-stage-to-orbit (SSTO) launch vehicle, (2) 'in-situ' resource utilization (ISRU)--specifically 'lunar-derived' liquid oxygen (LUNOX), and (3) LOX-augmented nuclear thermal rocket (LANTR) propulsion. Lunar transportation system elements consisting of a LANTR-powered lunar transfer vehicle (LTV) and a chemical propulsion lunar landing/Earth return vehicle (LERV) are configured to fit within the 'compact' dimensions of the SSTO cargo bay (diameter: 4.6 m/length: 9.0 m) while satisfying an initial mass in low Earth orbit (IMLEO) limit of approximately 60 t (3 SSTO launches). Using approximately 8 t of LUNOX to 'reoxidize' the LERV for a 'direct return' flight to Earth reduces its size and mass allowing delivery to LEO on a single 20 t SSTO launch. Similarly, the LANTR engine's ability to operate at any oxygen/ hydrogen mixture ratio from 0 to 7 with high specific impulse (approximately 940 to 515 s) is exploited to reduce hydrogen tank volume, thereby improving packaging of the LANTR LTV's 'propulsion' and 'propellant modules'. Expendable and reusable, piloted and cargo missions and vehicle designs are presented along with estimates of LUNOX production required to support the different mission modes. Concluding remarks address the issue of lunar transportation system costs from the launch vehicle perspective.

  20. Preliminary micro-Raman images of normal and malignant human skin cells

    NASA Astrophysics Data System (ADS)

    Short, Michael A.; Lui, Harvey; McLean, David I.; Zeng, Haishan; Chen, Michael X.

    2006-02-01

    Micro-Raman spectroscopy covering a frequency range from 200 to 4000 cm -1 was used to image human skin melanocytes and keratinocytes with a spatial resolution of 0.5 μm. The cells were either cultivated on glass microscope slides or were located within thin sections of skin biopsies mounted on low fluorescence BaF II. A commercially available system was used to obtain the spectra utilizing a x100 long working distance objective with a numerical aperture of 0.8, and a cooled CCD. Both 633 and 515 nm excitations were tried, although the latter proved to be more effcient at producing Raman emission mostly due to the 1/λ 4 dependence in light scattering. Fluorescence emission from the cells was surprisingly low. The excitation power at the sample was kept below about 2 mW to avoid damaging the cells; this was the limiting factor on how quickly a Raman image could be obtained. Despite this diffculty we were able to obtain Raman images with rich information about the spectroscopic and structural features within the cytoplasm and cell nuclei. Differences were observed between the Raman images of normal and malignant cells. Spectra from purified DNA, RNA, lipids, proteins and melanin were obtained and these spectra were compared with the skin cell spectra with the aim of understanding how they are distributed over a cell and how the distribution changes between different cells.

  1. Design and preliminary testing of a handheld antagonistic SMA actuator for cancellation of human tremor

    NASA Astrophysics Data System (ADS)

    Pathak, Anupam; Brei, Diann; Luntz, Jonathan

    2009-03-01

    Essential Tremor is a debilitating disorder that in the US alone is estimated to affect up to ten million people. Unfortunately current treatments (i.e. drug therapy and surgical procedures), are limited in effectiveness and often pose a risk of adverse side-effects. In response to this problem, this paper describes an active cancellation device based on a hand-held Shape Memory Alloy (SMA) actuated stabilization platform. The assistive device is designed to hold and stabilize various objects (e.g. eating utensils, tools, pointing implements, etc.) by sensing the user's tremor and moving the object in an opposite direction using SMA actuators configured in biologically inspired antagonistic pairs. To aid in the design, performance prediction and control of the device, a device model is described that accounts for the device kinematics, SMA thermo-mechanics, and the heat transfer resulting from electrical heating and convective cooling. The system of differential equations in this device model coupled with the controller gain can be utilized to design the operation given a frequency range and power requirement. To demonstrate this, a prototype was built and experimentally tested under external disturbances in the range of 1-5 Hz, resulting in amplitude reduction of up to 80%. The extent of cancellation measured for both single-frequencies and actual human tremor disturbances demonstrate the promise of this approach as a broadly used assistive device for the multitudes afflicted by tremor.

  2. Crystallization and preliminary X-ray diffraction analysis of human endoplasmic reticulum aminopeptidase 2.

    PubMed

    Ascher, David B; Polekhina, Galina; Parker, Michael W

    2012-04-01

    Endoplasmic reticulum aminopeptidase 2 (ERAP2) is a critical enzyme involved in the final processing of MHC class I antigens. Peptide trimming by ERAP2 and the other members of the oxytocinase subfamily is essential to customize longer precursor peptides in order to fit them to the correct length required for presentation on major histocompatibility complex class I molecules. While recent structures of ERAP1 have provided an understanding of the `molecular-ruler' mechanism of substrate selection, little is known about the complementary activities of its homologue ERAP2 despite their sharing 49% sequence identity. In order to gain insights into the structure-function relationship of the oxytocinase subfamily, and in particular ERAP2, the luminal region of human ERAP2 has been crystallized in the presence of the inhibitor bestatin. The crystals belonged to an orthorhombic space group and diffracted anisotropically to 3.3 Å resolution in the best direction on an in-house X-ray source. A molecular-replacement solution suggested that the enzyme has adopted the closed state as has been observed in other inhibitor-bound aminopeptidase structures.

  3. Interactions of human P-glycoprotein transport substrates and inhibitors at the drug binding domain: Functional and molecular docking analyses.

    PubMed

    Kadioglu, Onat; Saeed, Mohamed E M; Valoti, Massimo; Frosini, Maria; Sgaragli, Giampietro; Efferth, Thomas

    2016-03-15

    Rhodamine 123 (R123) transport substrate sensitizes P-glycoprotein (P-gp) to inhibition by compound 2c (cis-cis) N,N-bis(cyclohexanolamine)aryl ester isomer in a concentration-dependent manner in human MDR1-gene transfected mouse T-lymphoma L5178 cells as shown previously. By contrast, epirubicin (EPI) concentration changes left unaltered 2c IC50 values of EPI efflux. To clarify this discrepancy, defined molecular docking (DMD) analyses of 12 N,N-bis(cyclohexanolamine)aryl esters, the highly flexible aryl ester analog 4, and several P-gp substrate/non-substrate inhibitors were performed on human P-gp drug- or nucleotide-binding domains (DBD or NBD). DMD measurements yielded lowest binding energy (LBE, kcal/mol) values (mean ± SD) ranging from -11.8 ± 0.54 (valspodar) to -3.98 ± 0.01 (4). Lys234, Ser952 and Tyr953 residues formed H-bonds with most of the compounds. Only 2c docked also at ATP binding site (LBE value of -6.9 ± 0.30 kcal/mol). Inhibition of P-gp-mediated R123 efflux by 12 N,N-bis(cyclohexanolamine)aryl esters and 4 significantly correlated with LBE values. DMD analysis of EPI, (3)H-1EPI, (3)H-2EPI, (14)C-1EPI, (14)C-2EPI, R123 and 2c before and after previous docking of each of them indicated that pre-docking of either 2c or EPI significantly reduced LBE of both EPI and R123, and that of both (3)H-2EPI and (14)C-2EPI, respectively. Since the clusters of DBD amino acid residues interacting with EPI were different, if EPI docked alone or after pre-docking of EPI or 2c, the existence of alternative secondary binding site for EPI on P-gp is credible. In conclusion, 2c may allocate the drug-binding pocket and reduce strong binding of EPI and R123 in agreement with P-gp inhibition experiments, where 2c reduced efflux of EPI and R123.

  4. A new marker for breast cancer diagnosis, human epididymis protein 4: A preliminary study

    PubMed Central

    Gündüz, Umut Riza; Gunaldi, Meral; Isiksacan, Nilgun; Gündüz, Seyda; Okuturlar, Yildiz; Kocoglu, Hakan

    2016-01-01

    Breast cancer is the most frequently diagnosed cancer type in women. Tumor markers have been widely used for assessing the treatment response and early diagnosis of recurrence. Human epididymis protein 4 (HE4) is expressed in ductal carcinoma of the breast tissue; however, its serum levels and their diagnostic and prognostic potential in breast cancer have not been investigated, which was therefore the aim of the present study. The serum levels of HE4 were determined in 36 breast cancer patients, 11 ovarian cancer patients and 16 healthy volunteers. The association between clinicopathological characteristics of breast cancer and serum HE4 levels was investigated. A significant difference in the median serum levels of HE4 was identified between breast cancer patients, ovarian cancer patients and healthy volunteers (P=0.013). The cutoff value for the prediction of breast cancer was determined at >13.24 pmol/l for HE4, with a sensitivity of 61.11%, specificity of 68.75%, positive predictive value of 81.48%, negative predictive value of 44.0% and accuracy of 63.46%. Furthermore, a positive correlation between the serum levels of HE4 and cancer antigen 15–3 was determined (r=0.399, P=0.026). To the best of our knowledge, the present study was the first to determine the diagnostic value of serum HE4 for breast cancer. A significant elevation of serum HE4 levels in patients with breast cancer compared with that in healthy controls was identified. HE4 may serve as a novel biomarker for the diagnosis of breast cancer. PMID:27446579

  5. Measurement of liver iron overload by magnetic induction using a planar gradiometer: preliminary human results.

    PubMed

    Casañas, R; Scharfetter, H; Altes, A; Remacha, A; Sarda, P; Sierra, J; Merwa, R; Hollaus, K; Rosell, J

    2004-02-01

    The measurement of hepatic iron overload is of particular interest in cases of hereditary hemochromatosis or in patients subject to periodic blood transfusion. The measurement of plasma ferritin provides an indirect estimate but the usefulness of this method is limited by many common clinical conditions (inflammation, infection, etc). Liver biopsy provides the most quantitative direct measurement of iron content in the liver but the risk of the procedure limits its acceptability. This work studies the feasibility of a magnetic induction (MI) low-cost system to measure liver iron overload. The excitation magnetic field (B0, frequency: 28 kHz) was produced by a coil, the perturbation produced by the object (deltaB) was detected using a planar gradiometer. We measured ten patients and seven volunteers in supine and prone positions. Each subject was moved in a plane parallel to the gradiometer several times to estimate measurement repeatability. The real and imaginary parts of deltaB/B0 were measured. Plastic tanks filled with water, saline and ferric solutions were measured for calibration purposes. We used a finite element model to evaluate the experimental results. To estimate the iron content we used the ratio between the maximum values for real and imaginary parts of deltaB/B0 and the area formed by the Nyquist plot divided by the maximum imaginary part. Measurements in humans showed that the contribution of the permittivity is stronger than the contribution of the permeability produced by iron stores in the liver. Defined iron estimators show a limited correlation with expected iron content in patients (R < or = 0.56). A more precise control of geometry and position of the subjects and measurements at multiple frequencies would improve the method.

  6. Internal morphology of the nonsyndromic prematurely fused sagittal suture in the human skull--A preliminary micro-CT study.

    PubMed

    Nowaczewska, W; Ziółkowski, G; Dybała, B

    2015-10-01

    Although nonsyndromic craniosynostosis (NSC) of the sagittal suture is a well-known type of craniosynostosis, little is currently known about the internal morphology of this prematurely fused suture in modern humans. Recently, micro-computed tomography (micro-CT) has been applied as a new tool for the quantitative evaluation of cranial suture morphology. However, so far there are only a small number of reports concerning studies of the internal morphology of prematurely fused sagittal suture in humans using micro-CT. The primary aim of this study was to examine the internal morphology of a completely obliterated sagittal suture in NSC. Two modern human skulls were used in this study: a skull of a child (aged 10 ± 2.5 years) displaying NSC of the sagittal suture and a skull of an adult showing non-prematurely completely obliterated sagittal suture. Quantitative variables of the sagittal sutures were assessed using method proposed by the authors. Porosity, and relative thickness of three bone layers in two examined skulls (inner cortical, diploë and outer cortical) were analysed using micro-CT in three equal sections of the sagittal suture. In the case of the prematurely fused suture, there were statistically significant differences mainly in the mean values of the porosity, thickness and relative thickness of the diploë between the anterior part and the two other parts (central and posterior) of this suture. Significant differences were also observed in some of the analysed variables between the sections of the sagittal suture of the skull with NSC and the normal skull.

  7. [Survey of arsenic concentrations in Chinese herbal medicines (CHMs) and preliminary risk assessment of As in CHMs on human health].

    PubMed

    Liu, Xiao-Juan; Liu, Wen-Ju; Lin, Ai-Jun; Liu, Yun-Xia

    2010-12-01

    Characteristics and concentrations of As in Chinese Herbal Medicines (CHMs) from Hebei province (including samples collected from fields and from medicine markets) were investigated, and the ADI (allowable daily intake) standard value from FAO/WHO was used for preliminary assessment of As risk on human health. The results showed that the average of As concentrations in different roots which were used as medicines, ranged from 0.14 mg/kg to 0.54 mg/kg,while for shoots which were used as medicines, average value of As concentrations in Dendranthema morifolium was 1.42 mg/kg, and the range in other species was from 0.09 mg/kg to 0.27 mg/kg. Based on Green Standards of Medicinal Plants and Preparations for Foreign Trade and Economy (2.0 mg/kg), the percentage of samples with As concentrations exceeding standard was 3.27% (n = 153) in roots and 9.09% (n = 44) in shoots,and 4.57% (n = 197) in total CHMs samples. As concentrations in CHMs from fields ranged from 0.03 mg/kg to 0.73 mg/kg,which were all lower than 2.0 mg/kg. However, As concentrations in CHMs from markets ranged from 0.05 mg/kg to 7.05 mg/kg, and the concentrations of As in 6.36% of samples exceeded the limited standard. Arsenic concentrations in samples from markets were significantly higher than those from fields (p < 0.05). As concentrations in Compositae were distributed widely from 0.08 mg/kg to 7.05 mg/kg,and the average at 0.87 mg/kg,while the average of As concentrations in other six families were between 0.21 mg/kg and 0.41 mg/kg, and As levels in 75% of samples for each family were below 0.5 mg/kg. Compared to Compositae samples, the CHMs from other families have higher security. ADI regulated by FAO/WHO was used to assess As risk in CHMs on human health in this study as well. As daily intake through CHMs from markets varied from 0.90 microg/d to 19.7 microg/d, and percentages of the daily intake of As in CHMs to ADI were in the range of 0.70% to 15.4%, which indicated that As in CHMs from markets

  8. Crystallization and preliminary X-ray diffraction analyses of pseudechetoxin and pseudecin, two snake-venom cysteine-rich secretory proteins that target cyclic nucleotide-gated ion channels

    SciTech Connect

    Suzuki, Nobuhiro; Yamazaki, Yasuo; Fujimoto, Zui; Morita, Takashi; Mizuno, Hiroshi

    2005-08-01

    Crystals of pseudechetoxin and pseudecin, potent peptidic inhibitors of cyclic nucleotide-gated ion channels, have been prepared and X-ray diffraction data have been collected to 2.25 and 1.90 Å resolution, respectively. Cyclic nucleotide-gated (CNG) ion channels play pivotal roles in sensory transduction of retinal and olfactory neurons. The elapid snake toxins pseudechetoxin (PsTx) and pseudecin (Pdc) are the only known protein blockers of CNG channels. These toxins are structurally classified as cysteine-rich secretory proteins and exhibit structural features that are quite distinct from those of other known small peptidic channel blockers. This article describes the crystallization and preliminary X-ray diffraction analyses of these toxins. Crystals of PsTx belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 60.30, b = 61.59, c = 251.69 Å, and diffraction data were collected to 2.25 Å resolution. Crystals of Pdc also belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with similar unit-cell parameters a = 60.71, b = 61.67, c = 251.22 Å, and diffraction data were collected to 1.90 Å resolution.

  9. Preliminary characterization of diacylglycerol generation in human basophils: temporal relationship to histamine release and resolution of degranulation.

    PubMed

    Oriente, A; Hundley, T; Hubbard, W C; MacGlashan, D W

    1997-05-01

    Purified human basophils were examined for changes in diacylglycerol levels to determine whether the transient nature of a N-formyl-methionyl-leucyl-phenylalanine (fMLP) -stimulated elevation in membrane protein kinase C (PKC) activity could be explained by the transient production of diacylglycerol (DAG). In preliminary experiments total DAG levels were measured by the DAG kinase assay. Although elevations followed stimulation with 1 microM fMLP (basal levels of 15 pmol/10(6) basophils vs. 45 pmol/10(6) basophils at the 3-min time point), there were no detectable changes in the first 60 s of the reaction. Histamine release is typically complete by 30-45 s. Measurement of inositol trisphosphate indicated a rapid increase by 5 s of 2.5 pmol/10(6) basophils. If DAG were produced at similar levels, the DAG kinase assay would not have detected the elevation. Consequently, fMLP-stimulated basophils were examined for changes in 1-stearoyl, 2-arachidonoyl, 3-sn-glycerol (SA-DAG) and 1-oleoyl, 2-arachidonoyl, 3-sn-glycerol by GC-NICIMS (negative ion chemical ionization mass spectroscopy). A 5-s elevation in these two species averaged 2 pmol/10(6) basophils, consistent with the inositol trisphosphate levels and occurring during the period of histamine release. However, a much more pronounced second phase to the SA-DAG response also occurred, mirroring the total DAG levels. This second phase of the DAG response, either total or SA-DAG, was transient on a time scale temporally coincident with the appearance and resolution of degranulation sacs as measured by fluorescence microscopy. These data suggest that there is selective generation of DAG species in the early reaction and the later appearance of DAG may be related to the formation and resolution of granule structures that follow the secretion of histamine.

  10. Non-invasive assessment of human tumour hypoxia with 123I-iodoazomycin arabinoside: preliminary report of a clinical study.

    PubMed Central

    Parliament, M. B.; Chapman, J. D.; Urtasun, R. C.; McEwan, A. J.; Golberg, L.; Mercer, J. R.; Mannan, R. H.; Wiebe, L. I.

    1992-01-01

    Non-invasive predictive assays which can confirm the presence or absence of hypoxic cells in human tumours show promise for understanding the natural history of tumour oxygenation, and improving the selection of patient subsets for novel radiotherapeutic strategies. Sensitiser adducts have been proposed as markers for hypoxic cells. Misonidazole analogues radiolabelled with iodine-123 have been developed for the detection of tumour hypoxia using conventional nuclear medicine techniques. In this pilot study, we have investigated one such potential marker, 123I-iodoazomycin arabinoside (123I-IAZA). Patients with advanced malignancies have undergone planar and single-photon emission computed tomographic (SPECT) imaging after intravenous administration of 123I-IAZA. We have observed radiotracer avidity in three out of ten tumours studied to date. Normal tissue activity of variable extent was also seen in the thyroid and salivary glands, upper aerodigestive tract, liver, intestine, and urinary bladder. Quantitative analysis of those images showing radiotracer avidity revealed tumour/normal tissue (T/N) ratios of 2.3 (primary small cell lung carcinoma), 1.9 (primary malignant fibrous histiocytoma) and 3.2 (brain metastasis from small cell lung carcinoma) at 18-24 h post injection. These preliminary data suggest that the use of gamma-emitter labelled 2-nitroimidazoles as diagnostic radiopharmaceuticals is feasible and safe, and that metabolic binding of 123I-IAZA is observed in some, but not all tumours. The inference that tumour 123I-IAZA avidity could be a non-invasive measure of tumour hypoxia deserves independent confirmation with needle oximetry. Images p92-a Figure 2 PMID:1310253

  11. Safety, pharmacokinetics, and preliminary assessment of efficacy of mecasermin (recombinant human IGF-1) for the treatment of Rett syndrome

    PubMed Central

    Khwaja, Omar S.; Ho, Eugenia; Barnes, Katherine V.; O’Leary, Heather M.; Pereira, Luis M.; Finkelstein, Yaron; Nelson, Charles A.; Vogel-Farley, Vanessa; DeGregorio, Geneva; Holm, Ingrid A.; Khatwa, Umakanth; Kapur, Kush; Alexander, Mark E.; Finnegan, Deirdre M.; Cantwell, Nicole G.; Walco, Alexandra C.; Rappaport, Leonard; Gregas, Matt; Fichorova, Raina N.; Shannon, Michael W.; Sur, Mriganka; Kaufmann, Walter E.

    2014-01-01

    Rett syndrome (RTT) is a severe X-linked neurodevelopmental disorder mainly affecting females and is associated with mutations in MECP2, the gene encoding methyl CpG-binding protein 2. Mouse models suggest that recombinant human insulin-like growth factor 1 (IGF-1) (rhIGF1) (mecasermin) may improve many clinical features. We evaluated the safety, tolerability, and pharmacokinetic profiles of IGF-1 in 12 girls with MECP2 mutations (9 with RTT). In addition, we performed a preliminary assessment of efficacy using automated cardiorespiratory measures, EEG, a set of RTT-oriented clinical assessments, and two standardized behavioral questionnaires. This phase 1 trial included a 4-wk multiple ascending dose (MAD) (40–120 μg/kg twice daily) period and a 20-wk open-label extension (OLE) at the maximum dose. Twelve subjects completed the MAD and 10 the entire study, without evidence of hypoglycemia or serious adverse events. Mecasermin reached the CNS compartment as evidenced by the increase in cerebrospinal fluid IGF-1 levels at the end of the MAD. The drug followed nonlinear kinetics, with greater distribution in the peripheral compartment. Cardiorespiratory measures showed that apnea improved during the OLE. Some neurobehavioral parameters, specifically measures of anxiety and mood also improved during the OLE. These improvements in mood and anxiety scores were supported by reversal of right frontal alpha band asymmetry on EEG, an index of anxiety and depression. Our data indicate that IGF-1 is safe and well tolerated in girls with RTT and, as demonstrated in preclinical studies, ameliorates certain breathing and behavioral abnormalities. PMID:24623853

  12. Preliminary results on an x-direction apparent mass model of human body sitting in a cushioned, suspended seat

    NASA Astrophysics Data System (ADS)

    Stein, George Juraj; Múčka, Peter; Chmúrny, Rudolf

    2006-12-01

    For modelling purposes and for evaluation of driver's seat performance in the vertical direction various mechano-mathematical models of seated human body have been developed and standardised by the international organisation for standardisation. No such models currently exist for human body sitting in an upright or slightly inclined position in a cushioned "armchair" type seat upper part, mounted on a mechanical, pneumatic or other type vertical suspension system. The interaction with the steering wheel and/or pedals has to be taken into consideration, as well as the variable position of the upper part of the human body in respect to the cushioned back-support of a driver's seat (full back contact to no contact at all), as observed in real driving conditions. This complex problem has to be simplified first to arrive at a manageable simpler mechano-mathematical model which still reflects the main problem features. A simple linear model of the human body apparent mass in the x-direction was designed and analysed. The model accounts for the reaction from the steering wheel and contact with the cushioned back-support of the seat "armchair" part. Model parameters were identified on basis of laboratory measurements. Out of three possible variant the most appropriate was singled out. The proposed model describes the measured apparent mass curve, and also gives indicative prediction of vibration transmissibility across the fore-and-aft ( x-direction) suspension system, if mounted and enabled. The proposed model can be a starting point for a further research in this field.

  13. Drugs of abuse, cytostatic drugs and iodinated contrast media in tap water from the Madrid region (central Spain):A case study to analyse their occurrence and human health risk characterization.

    PubMed

    Mendoza, A; Zonja, B; Mastroianni, N; Negreira, N; López de Alda, M; Pérez, S; Barceló, D; Gil, A; Valcárcel, Y

    2016-01-01

    This work analyses the presence of forty-eight emerging pollutants, including twenty-five drugs of abuse and metabolites, seventeen cytostatic drugs and six iodinated contrast media, in tap water from the Madrid Region. Analysis of the target compounds in the tap water was performed by means of (on-line or off-line) solid-phase extraction followed by analysis by liquid chromatography-tandem mass spectrometry. A preliminary human health risk characterization was undertaken for each individual compound and for different groups of compounds with a common mechanism of action found in tap water. The results of the study showed the presence of eight out of the twenty-five drugs of abuse and metabolites analysed, namely, the cocainics cocaine and benzoylecgonine, the amphetamine-type stimulants ephedrine, 3,4-methylenedioxymethamphetamine and methamphetamine, the opioid methadone and its metabolite 2-ethylene-1,5-dimethyl-3,3-diphenylpyrrolidine and, finally caffeine at concentrations ranging from 0.11 to 502 ng L(-1). Four out of the six analysed iodinated contrast media, namely, diatrizoate, iohexol, iomeprol and iopromide, were detected in at least one sample, with concentration values varying between 0.4 and 5 ng L(-1). Cytostatic compounds were not detected in any sample. Caffeine was the substance showing the highest concentrations, up to 502 ng L(-1), mainly in the drinking water sampling point located in Madrid city. Among the other drugs of abuse, the most abundant compounds were cocaine and benzoylecgonine, detected at concentrations ranging from 0.11 to 86 ng L(-1) and from 0.11 to 53 ng L(-1), respectively. Regarding iodinated contrast media, iohexol was the most ubiquitous and abundant compound, with a frequency of detection of 100% and concentrations from 0.5 to 5.0 ng L(-1) in basically the same range in all sampling points. Taking into account the results and types of treatment applied, ozonisation plus granular activated carbon filtration appears to be

  14. Indications of human activity from amino acid and amino sugar analyses on Holocene sediments from lake Lonar, central India

    NASA Astrophysics Data System (ADS)

    Menzel, P.; Gaye, B.; Wiesner, M.; Prasad, S.; Basavaiah, N.; Stebich, M.; Anoop, A.; Riedel, N.; Brauer, A.

    2012-04-01

    The DFG funded HIMPAC (Himalaya: Modern and Past Climates) programme aims to reconstruct Holocene Indian Monsoon climate using a multi-proxy and multi-archive approach. First investigations made on sediments from a ca. 10 m long core covering the whole Holocene taken from the lake Lonar in central India's state Maharashtra, Buldhana District, serve to identify changes in sedimentation, lake chemistry, local vegetation and regional to supra-regional climate patterns. Lake Lonar occupies the floor of an impact crater that formed on the ~ 65 Ma old basalt flows of the Deccan Traps. It covers an area of ca. 1 km2 and is situated in India's core monsoon area. The modern lake has a maximum depth of about 5 m, is highly alkaline, and hyposaline, grouped in the Na-Cl-CO3 subtype of saline lakes. No out-flowing stream is present and only three small streams feed the lake, resulting in a lake level highly sensitive to precipitation and evaporation. The lake is eutrophic and stratified throughout most of the year with sub- to anoxic waters below 2 m depth. In this study the core sediments were analysed for their total amino acid (AA) and amino sugar (AS) content, the amino acid bound C and N percentage of organic C and total N in the sediment and the distribution of individual amino acids. The results roughly show three zones within the core separated by distinct changes in their AA content and distribution. (i) The bottom part of the core from ca. 12000 cal a BP to 11400 cal a BP with very low AA and AS percentage indicating high lithogenic contribution, most probably related to dry conditions. (ii) From 11400 cal a BP to 1200 cal a BP the sediments show moderate AA and AS percentages and low values for the ratios of proteinogenic AAs to their non-proteinogenic degradation products (e.g. ASP/β-ALA; GLU/γ-ABA). (iii) The top part of the core (< 1200 cal a BP) is characterised by an intense increase in total AA and AS, AA-C/Corg and AA-N/Ntotas well as in the ratio of

  15. Pioglitazone and bladder cancer in human studies: is it diabetes itself, diabetes drugs, flawed analyses or different ethnicities?

    PubMed

    Tseng, Chin-Hsiao

    2012-03-01

    This article reviews human observations on pioglitazone and bladder cancer risk. The PROspective pioglitAzone Clinical Trial In macroVascular Events trial showed an imbalance in bladder cancer between users of pioglitazone and placebo (14 versus six cases, p = 0.069). However, after excluding bladder cancer probably ascribed to other etiology, a blind assessment concluded that the imbalance might not be related to pioglitazone. Epidemiologic studies conducted in the United States and France using insurance databases independently suggested that pioglitazone use for >2 years might confer a 20%-40% higher risk. Another study evaluating bladder cancer risk in diabetic patients using the National Health Insurance in Taiwan did not find any incident bladder cancer case among 422 pioglitazone users for a follow-up of up to 3 years. Because observational studies may suffer from selection and information bias, and inadequate adjustment for confounders may inflate the estimated risk, causal inference from these studies should be interpreted with caution. While investigating cancer risk associated with a medication, indication bias should also be attended, especially when the medication is used at a late stage of the disease. Because pioglitazone is usually a second or third line antidiabetic agent, the users are always characterized by older age, longer diabetes duration, poorer glycemic control, and higher rates of complications and comorbidities. Biased estimates will also result if these differences are not appropriately addressed in the analyses. Current evidence neither concludes nor excludes a causal role of pioglitazone on bladder cancer. Clinical trials aiming at evaluating the risk of cancer associated with a medication is not ethical and may not be expected to provide an answer on the issue of pioglitazone-related bladder cancer. However, a meta-analysis using all available clinical trials to compare the bladder cancer risk between pioglitazone and comparators

  16. Purification, crystallization and preliminary X-ray diffraction of wild-type and mutant recombinant human transforming growth factor β-induced protein (TGFBIp)

    PubMed Central

    Runager, Kasper; García-Castellanos, Raquel; Valnickova, Zuzana; Kristensen, Torsten; Nielsen, Niels Chr.; Klintworth, Gordon K.; Gomis-Rüth, F. Xavier; Enghild, Jan J.

    2009-01-01

    Transforming growth factor β-induced protein (TGFBIp) has been linked to several corneal dystrophies as certain point mutations in the protein may give rise to a progressive accumulation of insoluble protein material in the human cornea. Little is known about the biological functions of this extracellular protein, which is expressed in various tissues throughout the human body. However, it has been found to interact with a number of extracellular matrix macromolecules such as collagens and proteoglycans. Structural information about TGFBIp might prove to be a valuable tool in the elucidation of its function and its role in corneal dystrophies caused by mutations in the TGFBI gene. A simple method for the purification of wild-type and mutant forms of recombinant human TGFBIp from human cells under native conditions is presented here. Moreover, the crystallization and preliminary X-ray analysis of TGFBIp are reported. PMID:19255489

  17. INCORPORATION OF HUMAN FACTORS ENGINEERING ANALYSES AND TOOLS INTO THE DESIGN PROCESS FOR DIGITAL CONTROL ROOM UPGRADES.

    SciTech Connect

    O'HARA,J.M.; BROWN,W.

    2004-09-19

    Many nuclear power plants are modernizing with digital instrumentation and control systems and computer-based human-system interfaces (HSIs). The purpose of this paper is to summarize the human factors engineering (HFE) activities that can help to ensure that the design meets personnel needs. HFE activities should be integrated into the design process as a regular part of the engineering effort of a plant modification. The HFE activities will help ensure that human performance issues are addressed, that new technology supports task performance, and that the HSIs are designed in a manner that is compatible with human physiological, cognitive and social characteristics.

  18. Integrative analyses of RNA editing, alternative splicing, and expression of young genes in human brain transcriptome by deep RNA sequencing.

    PubMed

    Wu, Dong-Dong; Ye, Ling-Qun; Li, Yan; Sun, Yan-Bo; Shao, Yi; Chen, Chunyan; Zhu, Zhu; Zhong, Li; Wang, Lu; Irwin, David M; Zhang, Yong E; Zhang, Ya-Ping

    2015-08-01

    Next-generation RNA sequencing has been successfully used for identification of transcript assembly, evaluation of gene expression levels, and detection of post-transcriptional modifications. Despite these large-scale studies, additional comprehensive RNA-seq data from different subregions of the human brain are required to fully evaluate the evolutionary patterns experienced by the human brain transcriptome. Here, we provide a total of 6.5 billion RNA-seq reads from different subregions of the human brain. A significant correlation was observed between the levels of alternative splicing and RNA editing, which might be explained by a competition between the molecular machineries responsible for the splicing and editing of RNA. Young human protein-coding genes demonstrate biased expression to the neocortical and non-neocortical regions during evolution on the lineage leading to humans. We also found that a significantly greater number of young human protein-coding genes are expressed in the putamen, a tissue that was also observed to have the highest level of RNA-editing activity. The putamen, which previously received little attention, plays an important role in cognitive ability, and our data suggest a potential contribution of the putamen to human evolution.

  19. AN EXAMPLE OF MODEL STRUCTURE DIFFERENCES USING SENSITIVITY ANALYSES IN PHYSIOLOGICALLY BASED PHARMACOKINETIC MODELS OF TRICHLOROETHYLENE IN HUMANS

    EPA Science Inventory

    Abstract Trichloroethylene (TCE) is an industrial chemical and an environmental contaminant. TCE and its metabolites may be carcinogenic and affect human health. Physiologically based pharmacokinetic (PBPK) models that differ in compartmentalization are developed for TCE metabo...

  20. Crystallization and preliminary X-ray diffraction studies of an RNA aptamer in complex with the human IgG Fc fragment

    PubMed Central

    Sugiyama, Shigeru; Nomura, Yusuke; Sakamoto, Taiichi; Kitatani, Tomoya; Kobayashi, Asako; Miyakawa, Shin; Takahashi, Yoshinori; Adachi, Hiroaki; Takano, Kazufumi; Murakami, Satoshi; Inoue, Tsuyoshi; Mori, Yusuke; Nakamura, Yoshikazu; Matsumura, Hiroyoshi

    2008-01-01

    Aptamers, which are folded DNA or RNA molecules, bind to target molecules with high affinity and specificity. An RNA aptamer specific for the Fc fragment of human immunoglobulin G (IgG) has recently been identified and it has been demonstrated that an optimized 24-nucleotide RNA aptamer binds to the Fc fragment of human IgG and not to other species. In order to clarify the structural basis of the high specificity of the RNA aptamer, it was crystallized in complex with the Fc fragment of human IgG1. Preliminary X-ray diffraction studies revealed that the crystals belonged to the orthorhombic space group P21212, with unit-cell parameters a = 83.7, b = 107.2, c = 79.0 Å. A data set has been collected to 2.2 Å resolution. PMID:18931441

  1. In Vivo Imaging of Human 11C-Metformin in Peripheral Organs: Dosimetry, Biodistribution, and Kinetic Analyses.

    PubMed

    Gormsen, Lars C; Sundelin, Elias Immanuel; Jensen, Jonas Brorson; Vendelbo, Mikkel Holm; Jakobsen, Steen; Munk, Ole Lajord; Hougaard Christensen, Mette Marie; Brøsen, Kim; Frøkiær, Jørgen; Jessen, Niels

    2016-12-01

    Metformin is the most widely prescribed oral antiglycemic drug, with few adverse effects. However, surprisingly little is known about its human biodistribution and target tissue metabolism. In animal experiments, we have shown that metformin can be labeled by (11)C and that (11)C-metformin PET can be used to measure renal function. Here, we extend these preclinical findings by a first-in-human (11)C-metformin PET dosimetry, biodistribution, and tissue kinetics study.

  2. Supplemental Performance Analyses for Igneous Activity and Human Intrusion at the Potential High-Level Nuclear Waste Repository at Yucca Mountain

    SciTech Connect

    Swift, P.; Gaither, K.; Freeze, G.; McCord, J.; Kalinich, D.; Saulnier, G.; Statham, W.

    2002-02-26

    The U.S. Department of Energy (DOE) is considering the possible recommendation of a site at Yucca Mountain, Nevada, for the potential development of a geologic repository for the disposal of high-level radioactive waste and spent nuclear fuel. Consequences of hypothetical disruption of the Yucca Mountain site by igneous activity or human intrusion have been evaluated in the Yucca Mountain Science and Engineering Report (S&ER) (1), which presents technical information supporting the consideration of the possible site recommendation. Since completion of the S&ER, supplemental analyses have examined possible impacts of new information and alternative assumptions on the estimates of the consequences of these events. Specifically, analyses of the consequences of igneous disruption address uncertainty regarding: (1) the impacts of changes in the repository footprint and waste package spacing on the probability of disruption; (2) impacts of alternative assumptions about the appropriat e distribution of future wind speeds to use in the analysis; (3) effects of alternative assumptions about waste particle sizes; and (4) alternative assumptions about the number of waste packages damaged by igneous intrusion; and (5) alternative assumptions about the exposure pathways and the biosphere dose conversion factors used in the analysis. Additional supplemental analyses, supporting the Final Environmental Impact Statement (FEIS), have examined the results for both igneous disruption and human intrusion, recalculated for a receptor group located 18 kilometers (km) from the repository (the location specified in 40 CFR 197), rather than at the 20 km distance used in the S&ER analyses.

  3. Clonal analyses and gene profiling identify genetic biomarkers of the thermogenic potential of human brown and white preadipocytes.

    PubMed

    Xue, Ruidan; Lynes, Matthew D; Dreyfuss, Jonathan M; Shamsi, Farnaz; Schulz, Tim J; Zhang, Hongbin; Huang, Tian Lian; Townsend, Kristy L; Li, Yiming; Takahashi, Hirokazu; Weiner, Lauren S; White, Andrew P; Lynes, Maureen S; Rubin, Lee L; Goodyear, Laurie J; Cypess, Aaron M; Tseng, Yu-Hua

    2015-07-01

    Targeting brown adipose tissue (BAT) content or activity has therapeutic potential for treating obesity and the metabolic syndrome by increasing energy expenditure. However, both inter- and intra-individual differences contribute to heterogeneity in human BAT and potentially to differential thermogenic capacity in human populations. Here we generated clones of brown and white preadipocytes from human neck fat and characterized their adipogenic and thermogenic differentiation. We combined an uncoupling protein 1 (UCP1) reporter system and expression profiling to define novel sets of gene signatures in human preadipocytes that could predict the thermogenic potential of the cells once they were maturated. Knocking out the positive UCP1 regulators, PREX1 and EDNRB, in brown preadipocytes using CRISPR-Cas9 markedly abolished the high level of UCP1 in brown adipocytes differentiated from the preadipocytes. Finally, we were able to prospectively isolate adipose progenitors with great thermogenic potential using the cell surface marker CD29. These data provide new insights into the cellular heterogeneity in human fat and offer potential biomarkers for identifying thermogenically competent preadipocytes.

  4. Multi-Scale Computational Analyses of JP-8 Fuel Droplets and Vapors in Human Respiratory Airway Models

    DTIC Science & Technology

    2007-10-31

    micron- and nano-size aerosols in representative human nasal airways; (ii) multi- component and/or impure droplet evaporation or hygroscopity; (iii...14 1.3.4 Evaporation and deposition of multi- component droplets...fuel surrogate in order to track at least 12 components , i.e., chemical markers, considered to be most harmful. 20080226474 (iv) Model development and

  5. Identification and Functional Analyses of 11 769 Full-length Human cDNAs Focused on Alternative Splicing

    PubMed Central

    Wakamatsu, Ai; Kimura, Kouichi; Yamamoto, Jun-ichi; Nishikawa, Tetsuo; Nomura, Nobuo; Sugano, Sumio; Isogai, Takao

    2009-01-01

    We analyzed diversity of mRNA produced as a result of alternative splicing in order to evaluate gene function. First, we predicted the number of human genes transcribed into protein-coding mRNAs by using the sequence information of full-length cDNAs and 5′-ESTs and obtained 23 241 of such human genes. Next, using these genes, we analyzed the mRNA diversity and consequently sequenced and identified 11 769 human full-length cDNAs whose predicted open reading frames were different from other known full-length cDNAs. Especially, 30% of the cDNAs we identified contained variation in the transcription start site (TSS). Our analysis, which particularly focused on multiple variable first exons (FEVs) formed due to the alternative utilization of TSSs, led to the identification of 261 FEVs expressed in the tissue-specific manner. Quantification of the expression profiles of 13 genes by real-time PCR analysis further confirmed the tissue-specific expression of FEVs, e.g. OXR1 had specific TSS in brain and tumor tissues, and so on. Finally, based on the results of our mRNA diversity analysis, we have created the FLJ Human cDNA Database. From our result, it has been understood mechanisms that one gene produces suitable protein-coding transcripts responding to the situation and the environment. PMID:19880432

  6. Personalized medicine in human space flight: using Omics based analyses to develop individualized countermeasures that enhance astronaut safety and performance.

    PubMed

    Schmidt, Michael A; Goodwin, Thomas J

    2013-01-01

    Space flight is one of the most extreme conditions encountered by humans. Advances in Omics methodologies (genomics, transcriptomics, proteomics, and metabolomics) have revealed that unique differences exist between individuals. These differences can be amplified in extreme conditions, such as space flight. A better understanding of individual differences may allow us to develop personalized countermeasure packages that optimize the safety and performance of each astronaut. In this review, we explore the role of "Omics" in advancing our ability to: (1) more thoroughly describe the biological response of humans in space; (2) describe molecular attributes of individual astronauts that alter the risk profile prior to entering the space environment; (3) deploy Omics techniques in the development of personalized countermeasures; and (4) develop a comprehensive Omics-based assessment and countermeasure platform that will guide human space flight in the future. In this review, we advance the concept of personalized medicine in human space flight, with the goal of enhancing astronaut safety and performance. Because the field is vast, we explore selected examples where biochemical individuality might significantly impact countermeasure development. These include gene and small molecule variants associated with: (1) metabolism of therapeutic drugs used in space; (2) one carbon metabolism and DNA stability; (3) iron metabolism, oxidative stress and damage, and DNA stability; and (4) essential input (Mg and Zn) effects on DNA repair. From these examples, we advance the case that widespread Omics profiling should serve as the foundation for aerospace medicine and research, explore methodological considerations to advance the field, and suggest why personalized medicine may become the standard of care for humans in space.

  7. Preliminary study of gaze toward humans in photographs by individuals with autism, Down syndrome, or other intellectual disabilities: implications for design of visual scene displays.

    PubMed

    Wilkinson, Krista M; Light, Janice

    2014-06-01

    Visual scene displays (VSDs) are a form of augmentative and alternative communication display in which language concepts are embedded into an image of a naturalistic event. VSDs are based on the theory that language learning occurs through interactions with other people, and recommendations for VSD design have emphasized using images of these events that include humans. However, many VSDs also include other items that could potentially be distracting. We examined gaze fixation in 18 school-aged participants with and without severe intellectual/developmental disabilities (i.e., individuals with typical development, autism, Down syndrome and other intellectual disabilities) while they viewed photographs with human figures of various sizes and locations in the image, appearing alongside other interesting, and potentially distracting items. In all groups, the human figures attracted attention rapidly (within 1.5 seconds). The proportions of each participant's own fixation time spent on the human figures were similar across all groups, as were the proportions of total fixations made to the human figures. Although the findings are preliminary, this initial evidence supports the inclusion of humans in VSD images.

  8. Performance characteristics of DRI, CEDIA, and REMEDi systems for preliminary tests of amphetamines and opiates in human urine.

    PubMed

    Huang, Min-Kun; Dai, Yu-Shan; Lee, Choung-Huei; Liu, Chiareiy; Tsay, Wen-Ing; Li, Jih-Heng

    2006-01-01

    Arrestee urine specimens (930) were tested with DRI, CEDIA, and REMEDi; those that tested positive for amphetamines and opiates (616 and 414, respectively) were then confirmed by gas chromatography-mass spectrometry. The performance characteristics of these three preliminary systems were evaluated using the following commonly used parameters: true positive, true negative, false positive, and false negative. The sensitivity, specificity, and efficiency of these methods were also calculated. Data derived from this study indicated DRI and CEDIA adapted by this study generated acceptable preliminary test results for amphetamine/methamphetamine and morphine/codeine, but not for MDA/MDMA and REMEDi has lower sensitivity than DRI and CEDIA, but with better specificity and efficiency, supporting its use under emergency room settings where drug concentrations in overdose cases are expectedly at high levels.

  9. Identification of Human N-Myristoylated Proteins from Human Complementary DNA Resources by Cell-Free and Cellular Metabolic Labeling Analyses

    PubMed Central

    Takamitsu, Emi; Otsuka, Motoaki; Haebara, Tatsuki; Yano, Manami; Matsuzaki, Kanako; Kobuchi, Hirotsugu; Moriya, Koko; Utsumi, Toshihiko

    2015-01-01

    To identify physiologically important human N-myristoylated proteins, 90 cDNA clones predicted to encode human N-myristoylated proteins were selected from a human cDNA resource (4,369 Kazusa ORFeome project human cDNA clones) by two bioinformatic N-myristoylation prediction systems, NMT-The MYR Predictor and Myristoylator. After database searches to exclude known human N-myristoylated proteins, 37 cDNA clones were selected as potential human N-myristoylated proteins. The susceptibility of these cDNA clones to protein N-myristoylation was first evaluated using fusion proteins in which the N-terminal ten amino acid residues were fused to an epitope-tagged model protein. Then, protein N-myristoylation of the gene products of full-length cDNAs was evaluated by metabolic labeling experiments both in an insect cell-free protein synthesis system and in transfected human cells. As a result, the products of 13 cDNA clones (FBXL7, PPM1B, SAMM50, PLEKHN, AIFM3, C22orf42, STK32A, FAM131C, DRICH1, MCC1, HID1, P2RX5, STK32B) were found to be human N-myristoylated proteins. Analysis of the role of protein N-myristoylation on the intracellular localization of SAMM50, a mitochondrial outer membrane protein, revealed that protein N-myristoylation was required for proper targeting of SAMM50 to mitochondria. Thus, the strategy used in this study is useful for the identification of physiologically important human N-myristoylated proteins from human cDNA resources. PMID:26308446

  10. Phylogenetic and phylodynamic analyses of human metapneumovirus in Buenos Aires (Argentina) for a three-year period (2009-2011).

    PubMed

    Velez Rueda, Ana Julia; Mistchenko, Alicia Susana; Viegas, Mariana

    2013-01-01

    Human metapneumovirus, which belongs to the Paramyxoviridae family and has been classified as a member of the Pneumovirus genus, is genetically and clinically similar to other family members such as human respiratory syncytial virus. A total of 1146 nasopharyngeal aspirates from pediatric patients with moderate and severe acute lower respiratory tract infections, hospitalized at the Ricardo Gutierrez Childreńs Hospital (Buenos Aires, Argentina), were tested by real time RT-PCR for human metapneumovirus. Results showed that 168 (14.65%) were positive. Thirty-six of these 168 samples were randomly selected to characterize positive cases molecularly. The phylogenetic analysis of the sequences of the G and F genes showed that genotypes A2 and B2 cocirculated during 2009 and 2010 and that only genotype A2 circulated in 2011 in Argentina. Genotype A2 prevailed during the study period, a fact supported by a higher effective population size (Neτ) and higher diversity as compared to that of genotype B2 (10.9% (SE 1.3%) vs. 1.7% (SE 0.4%), respectively). The phylogeographic analysis of the G protein gene sequences showed that this virus has no geographical restrictions and can travel globally harbored in hosts. The selection pressure analysis of the F protein showed that although this protein has regions with polymorphisms, it has vast structural and functional constraints. In addition, the predicted B-linear epitopes and the sites recognized by previously described monoclonal antibodies were conserved in all Argentine sequences. This points out this protein as a potential candidate to be the target of future humanized antibodies or vaccines.

  11. Integrative mRNA-microRNA analyses reveal novel interactions related to insulin sensitivity in human adipose tissue.

    PubMed

    Kirby, Tyler J; Walton, R Grace; Finlin, Brian; Zhu, Beibei; Unal, Resat; Rasouli, Neda; Peterson, Charlotte A; Kern, Philip A

    2016-02-01

    Adipose tissue has profound effects on whole-body insulin sensitivity. However, the underlying biological processes are quite complex and likely multifactorial. For instance, the adipose transcriptome is posttranscriptionally modulated by microRNAs, but the relationship between microRNAs and insulin sensitivity in humans remains to be determined. To this end, we utilized an integrative mRNA-microRNA microarray approach to identify putative molecular interactions that regulate the transcriptome in subcutaneous adipose tissue of insulin-sensitive (IS) and insulin-resistant (IR) individuals. Using the NanoString nCounter Human v1 microRNA Expression Assay, we show that 17 microRNAs are differentially expressed in IR vs. IS. Of these, 16 microRNAs (94%) are downregulated in IR vs. IS, including miR-26b, miR-30b, and miR-145. Using Agilent Human Whole Genome arrays, we identified genes that were predicted targets of miR-26b, miR-30b, and miR-145 and were upregulated in IR subjects. This analysis produced ADAM22, MYO5A, LOX, and GM2A as predicted gene targets of these microRNAs. We then validated that miR-145 and miR-30b regulate these mRNAs in differentiated human adipose stem cells. We suggest that use of bioinformatic integration of mRNA and microRNA arrays yields verifiable mRNA-microRNA pairs that are associated with insulin resistance and can be validated in vitro.

  12. Structure-Dependent Deconjugation of Flavonoid Glucuronides by Human β-Glucuronidase - In Vitro and In Silico Analyses.

    PubMed

    Untergehrer, Monika; Bücherl, Daniel; Wittmann, Hans-Joachim; Strasser, Andrea; Heilmann, Jörg; Jürgenliemk, Guido

    2015-08-01

    Flavonoid glycosides are extensively metabolized to glucuronidated compounds after oral intake. Recently, a cleavage of quercetin glucuronides by β-glucuronidase has been found. To characterize the deglucuronidation reaction and its structural prerequisites among the flavonoid subtypes more precisely, four flavonol glucuronides with varying glucuronidation positions, five flavone 7-O-glucuronides with varying A- and B-ring substitution as well as one flavanone- and one isoflavone-7-O-glucuronide were analyzed in a human monocytic cell line. Investigation of the deglucuronidation rates by HPLC revealed a significant influence of the glucuronidation position on enzyme activity for flavonols. Across the flavonoid subtypes, the C-ring saturation also showed a significant influence on deglucuronidation, whereas A- and B-ring variations within the flavone-7-O-glucuronides did not affect the enzymes' activity. Results were compared to computational binding studies on human β-glucuronidase. Additionally, molecular modeling and dynamic studies were performed to obtain detailed insight into the binding and cleavage mode of the substrate at the active site of the human β-glucuronidase.

  13. Ancient DNA analyses exclude humans as the driving force behind late Pleistocene musk ox (Ovibos moschatus) population dynamics.

    PubMed

    Campos, Paula F; Willerslev, Eske; Sher, Andrei; Orlando, Ludovic; Axelsson, Erik; Tikhonov, Alexei; Aaris-Sørensen, Kim; Greenwood, Alex D; Kahlke, Ralf-Dietrich; Kosintsev, Pavel; Krakhmalnaya, Tatiana; Kuznetsova, Tatyana; Lemey, Philippe; MacPhee, Ross; Norris, Christopher A; Shepherd, Kieran; Suchard, Marc A; Zazula, Grant D; Shapiro, Beth; Gilbert, M Thomas P

    2010-03-23

    The causes of the late Pleistocene megafaunal extinctions are poorly understood. Different lines of evidence point to climate change, the arrival of humans, or a combination of these events as the trigger. Although many species went extinct, others, such as caribou and bison, survived to the present. The musk ox has an intermediate story: relatively abundant during the Pleistocene, it is now restricted to Greenland and the Arctic Archipelago. In this study, we use ancient DNA sequences, temporally unbiased summary statistics, and Bayesian analytical techniques to infer musk ox population dynamics throughout the late Pleistocene and Holocene. Our results reveal that musk ox genetic diversity was much higher during the Pleistocene than at present, and has undergone several expansions and contractions over the past 60,000 years. Northeast Siberia was of key importance, as it was the geographic origin of all samples studied and held a large diverse population until local extinction at approximately 45,000 radiocarbon years before present ((14)C YBP). Subsequently, musk ox genetic diversity reincreased at ca. 30,000 (14)C YBP, recontracted at ca. 18,000 (14)C YBP, and finally recovered in the middle Holocene. The arrival of humans into relevant areas of the musk ox range did not affect their mitochondrial diversity, and both musk ox and humans expanded into Greenland concomitantly. Thus, their population dynamics are better explained by a nonanthropogenic cause (for example, environmental change), a hypothesis supported by historic observations on the sensitivity of the species to both climatic warming and fluctuations.

  14. Comprehensive Analyses of White-Handed Gibbon Chromosomes Enables Access to 92 Evolutionary Conserved Breakpoints Compared to the Human Genome.

    PubMed

    Weise, Anja; Kosyakova, Nadezda; Voigt, Martin; Aust, Nadine; Mrasek, Kristin; Löhmer, Sharon; Rubtsov, Nikolai; Karamysheva, Tatyana V; Trifonov, Vladimir A; Hardekopf, David; Jančušková, Tereza; Pekova, Sona; Wilhelm, Kathleen; Liehr, Thomas; Fan, Xiaobo

    2015-01-01

    Gibbon species (Hylobatidae) impress with an unusually high number of numerical and structural chromosomal changes within the family itself as well as compared to other Hominoidea including humans. In former studies applying molecular cytogenetic methods, 86 evolutionary conserved breakpoints (ECBs) were reported in the white-handed gibbon (Hylobates lar, HLA) with respect to the human genome. To analyze those ECBs in more detail and also to achieve a better understanding of the fast karyotype evolution in Hylobatidae, molecular data for these regions are indispensably necessary. In the present study, we obtained whole chromosome-specific probes by microdissection of all 21 HLA autosomes and prepared them for aCGH. Locus-specific DNA probes were also used for further molecular cytogenetic characterization of selected regions. Thus, we could map 6 yet unreported ECBs in HLA with respect to the human genome. Additionally, in 26 of the 86 previously reported ECBs, the present approach enabled a more precise breakpoint mapping. Interestingly, a preferred localization of ECBs within segmental duplications, copy number variant regions, and fragile sites was observed.

  15. Signalling protein complexes isolated from primary human skin-resident T cells can be analysed by Multiplex IP-FCM.

    PubMed

    Smith, Stephen E P; Neier, Steven C; Davis, Tessa R; Pittelkow, Mark R; Gil, Diana; Schrum, Adam G

    2014-04-01

    Studying signal transduction in skin-resident T cells (sr-T cells) can be limited by the small size of clinical biopsies. Here, we isolated sr-T cells from clinical samples and analysed signalling protein complexes by multiplex immunoprecipitation detected by flow cytometry (mIP-FCM). In samples from two independent donors, antigenic stimulation induced signalling proteins to join shared complexes that were observed in seven pairwise combinations among five proteins. This demonstrates that sr-T cells isolated from small clinical samples provide sufficient material for mIP-FCM-based analysis of signalling-induced protein complexes. We propose that this strategy may be useful for gaining improved mechanistic insight of sr-T cell signal transduction associated with dermatological disease.

  16. Does individual quality mask the detection of performance trade-offs? A test using analyses of human physical performance.

    PubMed

    Wilson, Robbie S; Niehaus, Amanda C; David, Gwendolyn; Hunter, Andrew; Smith, Michelle

    2014-02-15

    Why are performance trade-offs so rarely detected in animals when their underlying physiological basis seems so intuitive? One possibility is that individual variation in health, fitness, nutrition, development or genetics, or 'individual quality', makes some individuals better or worse performers across all motor tasks. If this is the case, then correcting for individual quality should reveal functional trade-offs that might otherwise be overlooked. We tested this idea by exploring trade-offs in maximum physical performance and motor skill function in semi-professional soccer players. We assessed individual performance across five maximum 'athletic' tasks providing independent measures of power, stamina and speed, as well as five soccer-specific 'motor skill' tasks providing independent measures of foot control. We expected to find functional trade-offs between pairs of traits (e.g. endurance versus power/speed tasks or specialist-generalist performance) - but only after correcting for individual quality. Analyses of standardised raw data found positive associations among several pairs of traits, but no evidence of performance trade-offs. Indeed, peak performance across a single athletic task (degree of specialisation) was positively associated with performance averaged across all other athletic tasks (generalist). However, after accounting for an individual's overall quality, several functional trade-offs became evident. Within our quality-corrected data, 1500 m-speed (endurance) was negatively associated with squat time (power), jump distance (power) and agility speed - reflecting the expected speed-endurance trade-off; and degree of specialisation was negatively associated with average performance across tasks. Taken together, our data support the idea that individual variation in general quality can mask the detection of performance trade-offs at the whole-animal level. These results highlight the possibility that studies may spuriously conclude certain

  17. Methods and theory in bone modeling drift: comparing spatial analyses of primary bone distributions in the human humerus.

    PubMed

    Maggiano, Corey M; Maggiano, Isabel S; Tiesler, Vera G; Chi-Keb, Julio R; Stout, Sam D

    2016-01-01

    This study compares two novel methods quantifying bone shaft tissue distributions, and relates observations on human humeral growth patterns for applications in anthropological and anatomical research. Microstructural variation in compact bone occurs due to developmental and mechanically adaptive circumstances that are 'recorded' by forming bone and are important for interpretations of growth, health, physical activity, adaptation, and identity in the past and present. Those interpretations hinge on a detailed understanding of the modeling process by which bones achieve their diametric shape, diaphyseal curvature, and general position relative to other elements. Bone modeling is a complex aspect of growth, potentially causing the shaft to drift transversely through formation and resorption on opposing cortices. Unfortunately, the specifics of modeling drift are largely unknown for most skeletal elements. Moreover, bone modeling has seen little quantitative methodological development compared with secondary bone processes, such as intracortical remodeling. The techniques proposed here, starburst point-count and 45° cross-polarization hand-drawn histomorphometry, permit the statistical and populational analysis of human primary tissue distributions and provide similar results despite being suitable for different applications. This analysis of a pooled archaeological and modern skeletal sample confirms the importance of extreme asymmetry in bone modeling as a major determinant of microstructural variation in diaphyses. Specifically, humeral drift is posteromedial in the human humerus, accompanied by a significant rotational trend. In general, results encourage the usage of endocortical primary bone distributions as an indicator and summary of bone modeling drift, enabling quantitative analysis by direction and proportion in other elements and populations.

  18. Insulin signaling in type 2 diabetes: experimental and modeling analyses reveal mechanisms of insulin resistance in human adipocytes.

    PubMed

    Brännmark, Cecilia; Nyman, Elin; Fagerholm, Siri; Bergenholm, Linnéa; Ekstrand, Eva-Maria; Cedersund, Gunnar; Strålfors, Peter

    2013-04-05

    Type 2 diabetes originates in an expanding adipose tissue that for unknown reasons becomes insulin resistant. Insulin resistance reflects impairments in insulin signaling, but mechanisms involved are unclear because current research is fragmented. We report a systems level mechanistic understanding of insulin resistance, using systems wide and internally consistent data from human adipocytes. Based on quantitative steady-state and dynamic time course data on signaling intermediaries, normally and in diabetes, we developed a dynamic mathematical model of insulin signaling. The model structure and parameters are identical in the normal and diabetic states of the model, except for three parameters that change in diabetes: (i) reduced concentration of insulin receptor, (ii) reduced concentration of insulin-regulated glucose transporter GLUT4, and (iii) changed feedback from mammalian target of rapamycin in complex with raptor (mTORC1). Modeling reveals that at the core of insulin resistance in human adipocytes is attenuation of a positive feedback from mTORC1 to the insulin receptor substrate-1, which explains reduced sensitivity and signal strength throughout the signaling network. Model simulations with inhibition of mTORC1 are comparable with experimental data on inhibition of mTORC1 using rapamycin in human adipocytes. We demonstrate the potential of the model for identification of drug targets, e.g. increasing the feedback restores insulin signaling, both at the cellular level and, using a multilevel model, at the whole body level. Our findings suggest that insulin resistance in an expanded adipose tissue results from cell growth restriction to prevent cell necrosis.

  19. A mathematical high bar-human body model for analysing and interpreting mechanical-energetic processes on the high bar.

    PubMed

    Arampatzis, A; Brüggemann, G P

    1998-12-01

    The aims of this study were: 1. To study the transfer of energy between the high bar and the gymnast. 2. To develop criteria from the utilisation of high bar elasticity and the utilisation of muscle capacity to assess the effectiveness of a movement solution. 3. To study the influence of varying segment movement upon release parameters. For these purposes a model of the human body attached to the high bar (high bar-human body model) was developed. The human body was modelled using a 15-segment body system. The joint-beam element method (superelement) was employed for modelling the high bar. A superelement consists of four rigid segments connected by joints (two Cardan joints and one rotational-translational joint) and springs (seven rotation springs and one tension-compression spring). The high bar was modelled using three superelements. The input data required for the high bar human body model were collected with video-kinematographic (50 Hz) and dynamometric (500 Hz) techniques. Masses and moments of inertia of the 15 segments were calculated using the data from the Zatsiorsky et al. (1984) model. There are two major phases characteristic of the giant swing prior to dismounts from the high bar. In the first phase the gymnast attempts to supply energy to the high bar-humanbody system through muscle activity and to store this energy in the high bar. The difference between the energy transferred to the high bar and the reduction in the total energy of the body could be adopted as a criterion for the utilisation of high bar elasticity. The energy previously transferred into the high bar is returned to the body during the second phase. An advantageous increase in total body energy at the end of the exercise could only be obtained through muscle energy supply. An index characterising the utilisation of muscle capacity was developed out of the difference between the increase in total body energy and the energy returned from the high bar. A delayed and initially slow but

  20. Biomedical Analyses of Mice Body Hair Exposed to Long-term Space Flight as a Compliment of Human Research

    NASA Astrophysics Data System (ADS)

    Mukai, Chiaki

    Introduction: To understand the effect of space environment characterized by microgravity and radiation on protein and mineral metabolisms is important for developing the countermeasures to the adverse effects happening on the astronauts who stay long-term in space. Thus JAXA has started a human research to study the effects of long-term exposure in space flight on gene expression and mineral metabolism by analyzing astronaut's hair grown in space since December 2009 (Experiment nicknamed "HAIR"). Ten human subjects who are the crew of the International Space Station (ISS) will be expected to complete this experiment. Thanks to the tissue sharing program of space-flown mice which is presented and organized by AGI(Italian Space Agency), we can also have an opportunity to analyze rodents samples which will greatly compliment human hair experiment by enable us to conduct more detailed analysis with the expansion of skin analysis which is not include in human experiment. The purpose of this flown-mice experiment is to study the effects of long-term exposure to space environment such as microgravity and space radiation on mineral and protein metabolism, the biological responses to the stress levels, and the initial process of skin carcinogenesis by analyzing hair shaft, its root cells, and skin. Approach and Method In this experiment, we analyzed hair shaft, hair root and skin. Hair samples with skin were taken from 3-month space-flown mice and ground-control mice in the AGI's tissue sharing program in 2009. The sample numbers of space-flown mice and control-mice were three and six, respectively. And they were at the Mice Drawer System (MDS) in ISS and in the laboratory of Geneva University. For the hair shaft, the mineral balance is investi-gated by energy dispersive X-ray spectroscopy (SEM-EDX). For hair root, the extracted RNA undergoes DNA microarray analysis, and will be further examined particular interests of gene-expression by real time Reverse Transcription

  1. Filamin C, a dysregulated protein in cancer revealed by label-free quantitative proteomic analyses of human gastric cancer cells.

    PubMed

    Qiao, Jie; Cui, Shu-Jian; Xu, Lei-Lei; Chen, Si-Jie; Yao, Jun; Jiang, Ying-Hua; Peng, Gang; Fang, Cai-Yun; Yang, Peng-Yuan; Liu, Feng

    2015-01-20

    Gastric cancer (GC) is the fourth and fifth most common cancer in men and women, respectively. We identified 2,750 proteins at false discovery rates of 1.3% (protein) and 0.03% (spectrum) by comparing the proteomic profiles of three GC and a normal gastric cell lines. Nine proteins were significantly dysregulated in all three GC cell lines, including filamin C, a muscle-specific filamin and a large actin-cross-linking protein. Downregulation of filamin C in GC cell lines and tissues were verified using quantitative PCR and immunohistochemistry. Data-mining using public microarray datasets shown that filamin C was significantly reduced in many human primary and metastasis cancers. Transient expression or silencing of filamin C affected the proliferation and colony formation of cancer cells. Silencing of endogenous filamin C enhanced cancer cell migration and invasion, whereas ectopic expression of filamin C had opposing effects. Silencing of filamin C increased the expression of matrix metallopeptidase 2 and improved the metastasis of prostate cancer in a zebrafish model. High filamin C associated with better prognosis of prostate cancer, leukemia and breast cancer patients. These findings establish a functional role of filamin C in human cancers and these data will be valuable for further study of its mechanisms.

  2. Gross alpha and beta activity analyses in urine-a routine laboratory method for internal human radioactivity detection.

    PubMed

    Chen, Xiaowen; Zhao, Luqian; Qin, Hongran; Zhao, Meijia; Zhou, Yirui; Yang, Shuqiang; Su, Xu; Xu, Xiaohua

    2014-05-01

    The aim of this work was to develop a method to provide rapid results for humans with internal radioactive contamination. The authors hypothesized that valuable information could be obtained from gas proportional counter techniques by screening urine samples from potentially exposed individuals rapidly. Recommended gross alpha and beta activity screening methods generally employ gas proportional counting techniques. Based on International Standards Organization (ISO) methods, improvements were made in the evaporation process to develop a method to provide rapid results, adequate sensitivity, and minimum sample preparation and operator intervention for humans with internal radioactive contamination. The method described by an American National Standards Institute publication was used to calibrate the gas proportional counter, and urine samples from patients with or without radionuclide treatment were measured to validate the method. By improving the evaporation process, the time required to perform the assay was reduced dramatically. Compared with the reference data, the results of the validation samples were very satisfactory with respect to gross-alpha and gross-beta activities. The gas flow proportional counting method described here has the potential for radioactivity monitoring in the body. This method was easy, efficient, and fast, and its application is of great utility in determining whether a sample should be analyzed by a more complicated method, for example radiochemical and/or γ-spectroscopy. In the future, it may be used commonly in medical examination and nuclear emergency treatment.Health Phys. 106(5):000-000; 2014.

  3. Postmortem Analyses Unveil the Poor Efficacy of Decontamination, Anti-Inflammatory and Immunosuppressive Therapies in Paraquat Human Intoxications

    PubMed Central

    Dinis-Oliveira, Ricardo Jorge; de Pinho, Paula Guedes; Santos, Liliana; Teixeira, Helena; Magalhães, Teresa; Santos, Agostinho; de Lourdes Bastos, Maria; Remião, Fernando; Duarte, José Alberto; Carvalho, Félix

    2009-01-01

    Background Fatalities resulting from paraquat (PQ) self-poisonings represent a major burden of this herbicide. Specific therapeutic approaches have been followed to interrupt its toxic pathway, namely decontamination measures to prevent PQ absorption and to increase its excretion from organism, as well as the administration of anti-inflammatory and immunosuppressive drugs. Until now, none of the postmortem studies resulting from human PQ poisonings have assessed the relationship of these therapeutic measures with PQ toxicokinetics and related histopathological lesions, these being the aims of the present study. Methodology/Principal Findings For that purpose, during 2008, we collected human fluids and tissues from five forensic autopsies following fatal PQ poisonings. PQ levels were measured by gas chromatography-ion trap mass spectrometry. Structural inflammatory lesions were evaluated by histological and immunohistochemistry analysis. The samples of cardiac blood, urine, gastric and duodenal wall, liver, lung, kidney, heart and diaphragm, showed quantifiable levels of PQ even at 6 days post-intoxication. Structural analysis showed diffused necrotic areas, intense macrophage activation and leukocyte infiltration in all analyzed tissues. By immunohistochemistry it was possible to observe a strong nuclear factor kappa-B (NF-κB) activation and excessive collagen deposition. Conclusions/Significance Considering the observed PQ levels in all analyzed tissues and the expressive inflammatory reaction that ultimately leads to fibrosis, we conclude that the therapeutic protocol usually performed needs to be reviewed, in order to increase the efficacy of PQ elimination from the body as well as to diminish the inflammatory process. PMID:19779613

  4. Analyses of germline, chromosomally integrated human herpesvirus 6A and B genomes indicate emergent infection and new inflammatory mediators.

    PubMed

    Tweedy, J; Spyrou, M A; Hubacek, P; Kuhl, U; Lassner, D; Gompels, U A

    2015-02-01

    Human herpesvirus-6A (HHV-6A) is rarer than HHV-6B in many infant populations. However, they are similarly prevalent as germline, chromosomally integrated genomes (ciHHV-6A/B). This integrated form affects 0.1-1 % of the human population, where potentially virus gene expression could be in every cell, although virus relationships and health effects are not clear. In a Czech/German patient cohort ciHHV-6A was more common and diverse than ciHHV-6B. Quantitative PCR, nucleotide sequencing and telomeric integration site amplification characterized ciHHV-6 in 44 German myocarditis/cardiomyopathy and Czech malignancy/inflammatory disease (MI) patients plus donors. Comparisons were made to sequences from global virus reference strains, and blood DNA from childhood-infections from Zambia (HHV-6A mainly) and Japan (HHV-6B). The MI cohort were 86 % (18/21) ciHHV-6A, the cardiac cohort 65 % (13/20) ciHHV-6B, suggesting different disease links. Reactivation was supported by findings of 1) recombination between ciHHV-6A and HHV-6B genes in 20 % (4/21) of the MI cohort; 2) expression in a patient subset, of early/late transcripts from the inflammatory mediator genes chemokine receptor U51 and chemokine U83, both identical to ciHHV-6A DNA sequences; and 3) superinfection shown by deep sequencing identifying minor virus-variants only in ciHHV-6A, which expressed transcripts, indicating virus infection reactivates latent ciHHV-6A. Half the MI cohort had more than two copies per cell, median 5.2, indicative of reactivation. Remarkably, the integrated genomes encoded the secreted-active form of virus chemokines, rare in virus from childhood-infections. This shows integrated virus genomes can contribute new human genes with links to inflammatory pathology and supports ciHHV-6A reactivation as a source for emergent infection.

  5. Cellular and molecular analyses of hprt mutation in human hepatocyte L02 cells after exposure to carbon ions

    NASA Astrophysics Data System (ADS)

    Li, Qiang; He, Jing; Jin, Xiao-Dong; Gong, Li; Li, Sha

    Mutations play an important role in carcinogenesis. The quantitative evaluation of mutation induction by heavy charged particles helps us to delineate the risks of space radiation on astronauts, as well as the risks of heavy ions on patients during tumor therapy. Hprt mutation assay, which has been used as a biological dosimeter, is an ideal gene mutation test in mammalian cells in vitro. In order to provide basic data and evidence for the risk assessment of heavy ions, the relationships between hprt mutation induction and radiation dose in human hepatocyte L02 cells were investigated for highand low-LET carbon ions and X-rays. Moreover, the carbon ion induced hprt mutation spectrum was analyzed. In our study, human hepatocyte L02 cells were irradiated with carbon ions with LET of 30keV/µm and X-rays (0.2keV/µm), respectively. The survival fraction of L02 cells was measured by means of colony-forming assay. The mutation frequency was detected by measuring 6-thioguanine-resistant clones after 10 days of incubation at the presence of 15mg/L 6-TG. To obtain the mutation spectrum, 9 10 mutant cell clones at each dose were randomly selected from the 6-TG containing medium, and were further cultured and analyzed. The deletion patterns of the 9 exons of hprt gene were analyzed with multiplex polymerase chain reactions (multiplex PCR). Our results show that the number of mutants per 106 surviving cells increased with increasing the radiation dose for both the irradiations, and the mutation frequency increased up to 1Gy while reduced with increasing dose further. Partial deletion was the most dominant deletion pattern in the hprt mutant cells, and with the increase of dose, hprt genes tended to have more total deletions and less point deletions. It can be inferred that human hepatocyte L02 cells are more radiosensitive to high-LET carbon ions than to low-LET X-rays, and carbon ions are more effective in inducing hprt mutation in L02 cells. It has been also found that the

  6. Overview of ozone human exposure and health risk analyses used in the U.S. EPA's review of the ozone air quality standard.

    SciTech Connect

    Whitfield, R. G.

    1999-03-04

    This paper presents an overview of the ozone human exposure and health risk analyses developed under sponsorship of the U.S. Environmental Protection Agency (EPA). These analyses are being used in the current review of the national ambient air quality standards (NAAQS) for ozone. The analyses consist of three principal steps: (1) estimating short-term ozone exposure for particular populations (exposure model); (2) estimating population response to exposures or concentrations (exposure-response or concentration-response models); and (3) integrating concentrations or exposure with concentration-response or exposure-response models to produce overall risk estimates (risk model). The exposure model, called the probabilistic NAAQS exposure model for ozone (pNEM/03), incorporates the following factors: hourly ambient ozone concentrations; spatial distribution of concentrations; ventilation state of individuals at time of exposure; and movement of people through various microenvironments (e.g., outdoors, indoors, inside a vehicle) of varying air quality. Exposure estimates are represented by probability distributions. Exposure-response relationships have been developed for several respiratory symptom and lung function health effects, based on the results of controlled human exposure studies. These relationships also are probabilistic and reflect uncertainties associated with sample size and variability of response among subjects. The analyses also provide estimates of excess hospital admissions in the New York City area based on results from an epidemiology study. Overall risk results for selected health endpoints and recently analyzed air quality scenarios associated with alternative 8-hour NAAQS and the current 1-hour standard for outdoor children are used to illustrate application of the methodology.

  7. Cryogenic propellant thermal control system design considerations, analyses, and concepts applied to a Mars human exploration mission

    NASA Technical Reports Server (NTRS)

    Plachta, David W.; Tucker, Stephen; Hoffman, David J.

    1993-01-01

    This paper analyzes, defines, and sizes cryogenic storage thermal control systems that meet the requirements of future NASA Mars human exploration missions. The design issues of this system include the projection of the existing Multilayer Insulation data base for cryogenic storage to much thicker (10 cm or more) insulation systems, the unknown heat leak from mechanical interfaces, and the thermal and structural performance effects of the large tank sizes required for a Mars mission. Acknowledging these unknown effects, heat loss projections are made based on extrapolation of the existing data base. The results indicate that hydrogen, methane, and oxygen are feasible propellants, and that the best suited thermal control sytems are 'thick' MLI, thermodynamic vent sytems, cryocoolers, and vacuum jackets.

  8. The Ups and Downs of Insulin Resistance and Type 2 Diabetes: Lessons from Genomic Analyses in Humans

    PubMed Central

    Sales, Vicencia; Patti, Mary-Elizabeth

    2012-01-01

    We are in the midst of a worldwide epidemic of type 2 diabetes (T2D) and obesity. Understanding the mechanisms underlying these diseases is critical if we are to halt their progression and ultimately prevent their development. The advent and widespread implementation of microarray technology has allowed analysis of small samples of human skeletal muscle, adipose, liver, pancreas and blood. While patterns differ in each tissue, several dominant themes have emerged from these studies, including altered expression of genes indicating increased inflammation and altered lipid and mitochondrial oxidative metabolism and insulin signaling in patients with T2D, and in some cases, in those at risk for disease. Unraveling which changes in gene expression are primary, and which are secondary to an insulin resistant or diabetes metabolic milieu remains a scientific challenge but we are one step closer. PMID:23459395

  9. Chemical Analyses

    NASA Technical Reports Server (NTRS)

    Bulluck, J. W.; Rushing, R. A.

    1994-01-01

    As a preliminary study on the effects of chemical aging of polymer materials MERL and TRI have examined two polymeric materials that are typically used for offshore umbilical applications. These two materials were Tefzel, a copolymer of ethylene and tetrafluoroethylene, and Coflon, polyvinylidene fluoride. The Coflon specimens were cut from pipe sections and exposed to H2S at various temperatures and pressures. One of these specimens was tested for methane permeation, and another for H2S permeation. The Tefzel specimens were cut from .05 mm sheet stock material and were exposed to methanol at elevated temperature and pressure. One of these specimens was exposed to methanol permeation for 2 days at 100 C and 2500 psi. An additional specimen was exposed to liquid methanol for 3 days at 150 C and 15 Bar. Virgin specimens of each material were similarly prepared and tested.

  10. Raman spectroscopic analyses of preserved historical specimens of human hair attributed to Robert Stephenson and Sir Isaac Newton.

    PubMed

    Edwards, Howell G M; Hassan, Nik F N; Wilson, Andrew S

    2004-10-01

    The Raman spectra of two historical specimens of human hair attributed to the engineer Robert Stephenson and scientist Sir Isaac Newton, preserved in private collections are reported. Comparisons are made with the Raman spectra of modern hair specimens and with hair from archaeological excavations. The hair spectra collected with a laser excitation of 785 nm are of a better quality than those collected using 1064 nm. The historical hair specimens are remarkably well-defined spectroscopically in terms of the amide I vibrational mode and the [small nu](SS), ascribed to a predominantly gauche-gauche-gauche CSSC conformation. The contrast with degraded hair specimens recovered from archaeological excavations is striking. The presence of a weak feature near 2590 cm(-1) in the hair samples attributed to a [small nu](SH) vibration could be indicative of a reduction process operative on the CSSC cystine keratotic linkages and a possible origin of this is bacterial biodegradation identified histologically. This study demonstrates the molecular information available from non-destructive Raman spectroscopic analysis from single hair shafts or small bundles of fibres which complements information available from histological and destructive analytical techniques for rare biological specimens subjected to conservation or curation procedures in museums or private collections.

  11. Comprehensive and quantitative proteomic analyses of zebrafish plasma reveals conserved protein profiles between genders and between zebrafish and human.

    PubMed

    Li, Caixia; Tan, Xing Fei; Lim, Teck Kwang; Lin, Qingsong; Gong, Zhiyuan

    2016-04-13

    Omic approaches have been increasingly used in the zebrafish model for holistic understanding of molecular events and mechanisms of tissue functions. However, plasma is rarely used for omic profiling because of the technical challenges in collecting sufficient blood. In this study, we employed two mass spectrometric (MS) approaches for a comprehensive characterization of zebrafish plasma proteome, i.e. conventional shotgun liquid chromatography-tandem mass spectrometry (LC-MS/MS) for an overview study and quantitative SWATH (Sequential Window Acquisition of all THeoretical fragment-ion spectra) for comparison between genders. 959 proteins were identified in the shotgun profiling with estimated concentrations spanning almost five orders of magnitudes. Other than the presence of a few highly abundant female egg yolk precursor proteins (vitellogenins), the proteomic profiles of male and female plasmas were very similar in both number and abundance and there were basically no other highly gender-biased proteins. The types of plasma proteins based on IPA (Ingenuity Pathway Analysis) classification and tissue sources of production were also very similar. Furthermore, the zebrafish plasma proteome shares significant similarities with human plasma proteome, in particular in top abundant proteins including apolipoproteins and complements. Thus, the current study provided a valuable dataset for future evaluation of plasma proteins in zebrafish.

  12. Physical human model eye and methods of its use to analyse optical performance of soft contact lenses.

    PubMed

    Bakaraju, Ravi C; Ehrmann, Klaus; Falk, Darrin; Ho, Arthur; Papas, Eric

    2010-08-02

    A bench-top physical model eye that closely replicates both anatomical and optical properties of an average human eye was designed and constructed. The cornea was sourced from a flouro-polymer with refractive index (RI) of 1.376 and crystalline lenses were made of Boston RGP polymers, EO and Equalens II, with an equivalent RI of 1.429 and 1.423 respectively. These materials served to make crystalline lens components of different age groups and accommodative states. De-Ionized water, with RI of 1.334 represented both aqueous and vitreous humor. The complementary metal-oxide sensor of a PixelLink digital camera with a resolution of 5MP and a 2.2 microm pixel pitch, hosted on a motor-base, served as the 'acting' retina. The translation and rotary functions of the motor-base facilitated the simulation of different states of ametropia and assessment of peripheral visual function, respectively. We validated one of its configurations to suit normal viewing conditions and results from the on and off-axis optical quality measurements are presented. As a demonstration of potential practical uses, several corrective soft contact lenses were placed on the model eye and their optical performance evaluated.

  13. Susceptibility of human immunodeficiency virus type 1 group O isolates to antiretroviral agents: in vitro phenotypic and genotypic analyses.

    PubMed Central

    Descamps, D; Collin, G; Letourneur, F; Apetrei, C; Damond, F; Loussert-Ajaka, I; Simon, F; Saragosti, S; Brun-Vézinet, F

    1997-01-01

    We investigated the phenotypic and genotypic susceptibility of 11 human immunodeficiency virus type 1 (HIV-1) group O strains to nucleoside and nonnucleoside reverse transcriptase (RT) inhibitors and protease inhibitors in vitro. Phenotypic susceptibility was determined by using a standardized in vitro assay of RT inhibition, taking into account the replication kinetics of each strain. HIV-1 group M and HIV-2 isolates were used as references. DNA from cocultured peripheral blood mononuclear cells was amplified by using pol-specific group O primers and cloned for sequencing. Group O isolates were highly sensitive to nucleoside inhibitors, but six isolates were naturally highly resistant to all of the nonnucleoside RT inhibitors tested. Phylogenetic analysis of the pol gene showed that these isolates formed a separate cluster within group O, and genotypic analysis revealed a tyrosine-to-cysteine substitution at residue 181. Differences in susceptibility to saquinavir and ritonavir (RTV) were not significant between group O and group M isolates, although the 50% inhibitory concentration of RTV for group O isolates was higher than that for the HIV-1 subtype B strains. The study of HIV-1 group O susceptibility to antiretroviral drugs revealed that the viruses tested had specific phenotypic characteristics contrasting with the group M phenotypic expression. PMID:9343254

  14. Phylogenetic and functional analyses of a plant protein related to human B-cell receptor-associated proteins.

    PubMed

    Atabekova, Anastasia K; Pankratenko, Anna V; Makarova, Svetlana S; Lazareva, Ekaterina A; Owens, Robert A; Solovyev, Andrey G; Morozov, Sergey Y

    2017-01-01

    Human B-cell receptor-associated protein BAP31 (HsBAP31) is the endoplasmic reticulum-resident protein involved in protein sorting and transport as well as pro-apoptotic signaling. Plant orthologs of HsBAP31 termed 'plant BAP-like proteins' (PBL proteins) have thus far remained unstudied. Recently, the PBL protein from Nicotiana tabacum (NtPBL) was identified as an interactor of Nt-4/1, a plant protein known to interact with plant virus movement proteins and affect the long-distance transport of potato spindle tuber viroid (PSTVd) via the phloem. Here, we have compared the sequences of PBL proteins and studied the biochemical properties of NtPBL. Analysis of a number of fully sequenced plant genomes revealed that PBL-encoding genes represent a small multigene family with up to six members per genome. Two conserved motifs were identified in the C-terminal region of PBL proteins. The NtPBL C-terminal hydrophilic region (NtPBL-C) was expressed in bacterial cells, purified, and used for analysis of its RNA binding properties in vitro. In gel shift experiments, NtPBL-C was found to bind several tested RNAs, showing the most efficient binding to microRNA precursors (pre-miRNA) and less efficient interaction with PSTVd. Mutational analysis suggested that NtPBL-C has a composite RNA-binding site, with two conserved lysine residues in the most C-terminal protein region being involved in binding of pre-miRNA but not PSTVd RNA. Virus-mediated transient expression of NtPBL-C in plants resulted in stunting and leaf malformation, developmental abnormalities similar to those described previously for blockage of miRNA biogenesis/function. We hypothesize that the NtPBL protein represents a previously undiscovered component of the miRNA pathway.

  15. Analyses of robotic traverses and sample sites in the Schrödinger basin for the HERACLES human-assisted sample return mission concept

    NASA Astrophysics Data System (ADS)

    Steenstra, Edgar S.; Martin, Dayl J. P.; McDonald, Francesca E.; Paisarnsombat, Sarinya; Venturino, Christian; O'Hara, Sean; Calzada-Diaz, Abigail; Bottoms, Shelby; Leader, Mark K.; Klaus, Kurt K.; van Westrenen, Wim; Needham, Debra H.; Kring, David A.

    2016-09-01

    The International Space Exploration Coordination Group (ISECG) developed an integrated Global Exploration Roadmap (GER) that outlines plans for human-assisted sample return from the lunar surface in ∼2024 and for human presence on the lunar surface in ∼2028. Previous studies have identified the Schrödinger basin, situated on the far side of the Moon, as a prime target for lunar science and exploration where a significant number of the scientific concepts reviewed by the National Research Council (NRC, 2007) can be addressed. In this study, two robotic mission traverses within the Schrödinger basin are proposed based on a 3 year mission plan in support of the HERACLES human-assisted sample return mission concept. A comprehensive set of modern remote sensing data (LROC imagery, LOLA topography, M3 and Clementine spectral data) has been integrated to provide high-resolution coverage of the traverses and to facilitate identification of specific sample localities. We also present a preliminary Concept of Operations (ConOps) study based on a set of notional rover capabilities and instrumental payload. An extended robotic mission to the Schrödinger basin will allow for significant sample return opportunities from multiple distinct geologic terrains and will address multiple high-priority NRC (2007) scientific objectives. Both traverses will offer the first opportunity to (i) sample pyroclastic material from the lunar farside, (ii) sample Schrödinger impact melt and test the lunar cataclysm hypothesis, (iii) sample deep crustal lithologies in an uplifted peak ring and test the lunar magma ocean hypothesis and (iv) explore the top of an impact melt sheet, enhancing our ability to interpret Apollo samples. The shorter traverse will provide the first opportunity to sample farside mare deposits, whereas the longer traverse has significant potential to collect SPA impact melt, which can be used to constrain the basin-forming epoch. These robotic missions will revalidate

  16. Multilayer-omics analyses of human cancers: exploration of biomarkers and drug targets based on the activities of the International Human Epigenome Consortium.

    PubMed

    Kanai, Yae; Arai, Eri

    2014-01-01

    Epigenetic alterations consisting mainly of DNA methylation alterations and histone modification alterations are frequently observed in cancers associated with chronic inflammation and/or persistent infection with viruses or other pathogenic microorganisms, or with cigarette smoking. Accumulating evidence suggests that alterations of DNA methylation are involved even in the early and precancerous stages. On the other hand, in patients with cancers, aberrant DNA methylation is frequently associated with tumor aggressiveness and poor patient outcome. Recently, epigenome alterations have been attracting a great deal of attention from researchers who are focusing on not only cancers but also neuronal, immune and metabolic disorders. In order to accurately identify disease-specific epigenome profiles that could be potentially applicable for disease prevention, diagnosis and therapy, strict comparison with standard epigenome profiles of normal tissues is indispensable. However, epigenome mechanisms show heterogeneity among tissues and cell lineages. Therefore, it is not easy to obtain a comprehensive picture of standard epigenome profiles of normal tissues. In 2010, the International Human Epigenome Consortium (IHEC) was established to coordinate the production of reference maps of human epigenomes for key cellular states. In order to gain substantial coverage of the human epigenome, the IHEC has set an ambitious goal to decipher at least 1000 epigenomes within the next 7-10 years. We consider that pathway analysis using genes showing multilayer-omics abnormalities, including genome, epigenome, transcriptome, proteome and metabolome abnormalities, may be useful for elucidating the molecular background of pathogenesis and for exploring possible therapeutic targets for each disease.

  17. Sociopolitical Analyses.

    ERIC Educational Resources Information Center

    Van Galen, Jane, Ed.; And Others

    1992-01-01

    This theme issue of the serial "Educational Foundations" contains four articles devoted to the topic of "Sociopolitical Analyses." In "An Interview with Peter L. McLaren," Mary Leach presented the views of Peter L. McLaren on topics of local and national discourses, values, and the politics of difference. Landon E.…

  18. Fasciola hepatica phenotypic characterization in Andean human endemic areas: valley versus altiplanic patterns analysed in liver flukes from sheep from Cajamarca and Mantaro, Peru.

    PubMed

    Valero, M Adela; Perez-Crespo, Ignácio; Khoubbane, Messaoud; Artigas, Patricio; Panova, Miroslava; Ortiz, Pedro; Maco, Vicente; Espinoza, José R; Mas-Coma, Santiago

    2012-03-01

    Fascioliasis is a zoonotic parasitic disease caused by Fasciola hepatica and Fasciola gigantica. Of both species, F. hepatica is the only one described in the Americas, mainly transmitted by lymnaeid snail vectors of the Galba/Fossaria group. Human fascioliasis endemic areas are mainly located in high altitude areas of Andean countries. Given the necessity to characterize F. hepatica populations involved, the phenotypic features of fasciolid adults infecting sheep present in human fascioliasis endemic areas were analysed in the Cajamarca Valley and Mantaro Valley (valley transmission patterns) and the northern Bolivian Altiplano (altiplanic transmission pattern). A computer image analysis system (CIAS) was applied on the basis of standardized measurements. The aforementioned highland populations were compared to standard lowland natural and experimental populations of European origin. Liver fluke size was studied by multivariate analyses. Two phenotypic patterns could be distinguished in F. hepatica adult size: the valley pattern (Cajamarca and Mantaro, Peru) and the altiplanic pattern (northern Altiplano, Bolivia). Results showed that the Andean valley population and European standard populations presented a phenotypic homogeneity. The Altiplano population showed a large size range with a pronouncedly lower minimum size indicating that uterus gravidity is reached at a smaller size than in valley populations. The results of this study demonstrate that there is no apparent relationship between the shape of fasciolid adults with regard to altitudinal difference or geographical origin and that allometry-free shape appears as a more stable trait than size in fasciolid species. Results are analysed in terms of intensity/crowding effect aspects and permanent/seasonal transmission characteristics.

  19. Crystallization and Preliminary X-ray Analysis of the Human Long Myosin Light-Chain Kinase 1-Specific Domain IgCAM3

    SciTech Connect

    W Vallen Graham; A Magis; K Bailey; J Turner; D Ostrov

    2011-12-31

    Myosin light-chain kinase-dependent tight junction regulation is a critical event in inflammatory cytokine-induced increases in epithelial paracellular permeability. MLCK is expressed in human intestinal epithelium as two isoforms, long MLCK1 and long MLCK2, and MLCK1 is specifically localized to the tight junction, where it regulates paracellular permeability. The sole difference between these long MLCK splice variants is the presence of an immunoglobulin-like cell-adhesion molecule domain, IgCAM3, in MLCK1. To gain insight into the structure of the IgCAM3 domain, the IgCAM3 domain of MLCK1 has been expressed, purified and crystallized. Preliminary X-ray diffraction data were collected to 2.0 {angstrom} resolution and were consistent with the primitive trigonal space group P2{sub 1}2{sub 1}2{sub 1}.

  20. Clastogenic effect of bracken fern (Pteridium aquilinum v. arachnoideum) diet in peripheral lymphocytes of human consumers: preliminary data.

    PubMed

    Recouso, R C; Stocco dos Santos, R C; Freitas, R; Santos, R C; de Freitas, A C; Brunner, O; Beçak, W; Lindsey, C J

    2003-03-01

    Ingestion of bracken fern (Pteridium aquilinum v. arachnoideum) is associated with digestive tract cancer in different regions of Japan, Venezuela and Brazil. In view of reports that dietary bracken fern causes chromosomal instability in cattle, the clastogenic effect of bracken fern was investigated, in a preliminary study, in peripheral lymphocytes obtained from habitual consumers and a control group of non-consumers, which were carefully investigated about cancer history or family cancer history, negative in both cases, using protocols comparable to those previously described in studies in cattle raised on bracken pastures. Cytogenetic analysis showed significant increased levels of chromosomal abnormalities, such as chromatid breaks, in cultured peripheral lymphocytes of the consumer group. There was no correlation with subjects, gender, smoking habits or alcohol consumption, and the only correlation was with prolonged exposure to dietary bracken.

  1. Analysing inter-relationships among water, governance, human development variables in developing countries: WatSan4Dev database coherency analysis

    NASA Astrophysics Data System (ADS)

    Dondeynaz, C.; Carmona Moreno, C.; Céspedes Lorente, J. J.

    2012-01-01

    The "Integrated Water Resources Management" principle was formally laid down at the International Conference on Water and Sustainable development in Dublin 1992. One of the main results of this conference is that improving Water and Sanitation Services (WSS), being a complex and interdisciplinary issue, passes through collaboration and coordination of different sectors (environment, health, economic activities, governance, and international cooperation). These sectors influence or are influenced by the access to WSS. The understanding of these interrelations appears as crucial for decision makers in the water sector. In this framework, the Joint Research Centre (JRC) of the European Commission (EC) has developed a new database (WatSan4Dev database) containing 45 indicators (called variables in this paper) from environmental, socio-economic, governance and financial aid flows data in developing countries. This paper describes the development of the WatSan4Dev dataset, the statistical processes needed to improve the data quality; and, finally, the analysis to verify the database coherence is presented. At the light of the first analysis, WatSan4Dev Dataset shows the coherency among the different variables that are confirmed by the direct field experience and/or the scientific literature in the domain. Preliminary analysis of the relationships indicates that the informal urbanisation development is an important factor influencing negatively the percentage of the population having access to WSS. Health, and in particular children health, benefits from the improvement of WSS. Efficient environmental governance is also an important factor for providing improved water supply services. The database would be at the base of posterior analyses to better understand the interrelationships between the different indicators associated in the water sector in developing countries. A data model using the different indicators will be realised on the next phase of this research work.

  2. Combined magnetic resonance and diffusion tensor imaging analyses provide a powerful tool for in vivo assessment of deformation along human muscle fibers.

    PubMed

    Pamuk, Uluç; Karakuzu, Agah; Ozturk, Cengizhan; Acar, Burak; Yucesoy, Can A

    2016-10-01

    Muscle fiber direction strain provides invaluable information for characterizing muscle function. However, methods to study this for human muscles in vivo are lacking. Using magnetic resonance (MR) imaging based deformation analyses and diffusion tensor (DT) imaging based tractography combined, we aimed to assess muscle fiber direction local tissue deformations within the human medial gastrocnemius (GM) muscle. Healthy female subjects (n=5, age=27±1 years) were positioned prone within the MR scanner in a relaxed state with the ankle angle fixed at 90°. The knee was brought to flexion (140.8±3.0°) (undeformed state). Sets of 3D high resolution MR, and DT images were acquired. This protocol was repeated at extended knee joint position (177.0±1.0°) (deformed state). Tractography and Demons nonrigid registration algorithm was utilized to calculate local deformations along muscle fascicles. Undeformed state images were also transformed by a synthetic rigid body motion to calculate strain errors. Mean strain errors were significantly smaller then mean fiber direction strains (lengthening: 0.2±0.1% vs. 8.7±8.5%; shortening: 3.3±0.9% vs. 7.5±4.6%). Shortening and lengthening (up to 23.3% and 116.7%, respectively) occurs simultaneously along individual fascicles despite imposed GM lengthening. Along-fiber shear strains confirm the presence of much shearing between fascicles. Mean fiber direction strains of different tracts also show non-uniform distribution. Inhomogeneity of fiber strain indicates epimuscular myofascial force transmission. We conclude that MR and DT imaging analyses combined provide a powerful tool for quantifying deformation along human muscle fibers in vivo. This can help substantially achieving a better understanding of normal and pathological muscle function and mechanisms of treatment techniques.

  3. Crystallization and preliminary X-ray analysis of a native human tRNA synthetase whose allelic variants are associated with Charcot–Marie–Tooth disease

    SciTech Connect

    Xie, Wei; Schimmel, Paul; Yang, Xiang-Lei

    2006-12-01

    Crystallization and preliminary X-ray analysis of a native human tRNA synthetase whose allelic variants are associated with Charcot–Marie–Tooth Disease. Glycyl-tRNA synthetase (GlyRS) is one of a group of enzymes that catalyze the synthesis of aminoacyl-tRNAs for translation. Mutations of human and mouse GlyRSs are causally associated with Charcot–Marie–Tooth disease, the most common genetic disorder of the peripheral nervous system. As the first step towards a structure–function analysis of this disease, native human GlyRS was expressed, purified and crystallized. The crystal belonged to space group P4{sub 3}2{sub 1}2 or its enantiomorphic space group P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 91.74, c = 247.18 Å, and diffracted X-rays to 3.0 Å resolution. The asymmetric unit contained one GlyRS molecule and had a solvent content of 69%.

  4. Targeting breast cancer stem cells by dendritic cell vaccination in humanized mice with breast tumor: preliminary results

    PubMed Central

    Pham, Phuc Van; Le, Hanh Thi; Vu, Binh Thanh; Pham, Viet Quoc; Le, Phong Minh; Phan, Nhan Lu-Chinh; Trinh, Ngu Van; Nguyen, Huyen Thi-Lam; Nguyen, Sinh Truong; Nguyen, Toan Linh; Phan, Ngoc Kim

    2016-01-01

    Background Breast cancer (BC) is one of the leading cancers in women. Recent progress has enabled BC to be cured with high efficiency. However, late detection or metastatic disease often renders the disease untreatable. Additionally, relapse is the main cause of death in BC patients. Breast cancer stem cells (BCSCs) are considered to cause the development of BC and are thought to be responsible for metastasis and relapse. This study aimed to target BCSCs using dendritic cells (DCs) to treat tumor-bearing humanized mice models. Materials and methods NOD/SCID mice were used to produce the humanized mice by transplantation of human hematopoietic stem cells. Human BCSCs were injected into the mammary fat pad to produce BC humanized mice. Both hematopoietic stem cells and DCs were isolated from the human umbilical cord blood, and immature DCs were produced from cultured mononuclear cells. DCs were matured by BCSC-derived antigen incubation for 48 hours. Mature DCs were vaccinated to BC humanized mice with a dose of 106 cells/mice, and the survival percentage was monitored in both treated and untreated groups. Results The results showed that DC vaccination could target BCSCs and reduce the tumor size and prolong survival. Conclusion These results suggested that targeting BCSCs with DCs is a promising therapy for BC. PMID:27499638

  5. MRI-derived measurements of human subcortical, ventricular and intracranial brain volumes: Reliability effects of scan sessions, acquisition sequences, data analyses, scanner upgrade, scanner vendors and field strengths.

    PubMed

    Jovicich, Jorge; Czanner, Silvester; Han, Xiao; Salat, David; van der Kouwe, Andre; Quinn, Brian; Pacheco, Jenni; Albert, Marilyn; Killiany, Ronald; Blacker, Deborah; Maguire, Paul; Rosas, Diana; Makris, Nikos; Gollub, Randy; Dale, Anders; Dickerson, Bradford C; Fischl, Bruce

    2009-05-15

    Automated MRI-derived measurements of in-vivo human brain volumes provide novel insights into normal and abnormal neuroanatomy, but little is known about measurement reliability. Here we assess the impact of image acquisition variables (scan session, MRI sequence, scanner upgrade, vendor and field strengths), FreeSurfer segmentation pre-processing variables (image averaging, B1 field inhomogeneity correction) and segmentation analysis variables (probabilistic atlas) on resultant image segmentation volumes from older (n=15, mean age 69.5) and younger (both n=5, mean ages 34 and 36.5) healthy subjects. The variability between hippocampal, thalamic, caudate, putamen, lateral ventricular and total intracranial volume measures across sessions on the same scanner on different days is less than 4.3% for the older group and less than 2.3% for the younger group. Within-scanner measurements are remarkably reliable across scan sessions, being minimally affected by averaging of multiple acquisitions, B1 correction, acquisition sequence (MPRAGE vs. multi-echo-FLASH), major scanner upgrades (Sonata-Avanto, Trio-TrioTIM), and segmentation atlas (MPRAGE or multi-echo-FLASH). Volume measurements across platforms (Siemens Sonata vs. GE Signa) and field strengths (1.5 T vs. 3 T) result in a volume difference bias but with a comparable variance as that measured within-scanner, implying that multi-site studies may not necessarily require a much larger sample to detect a specific effect. These results suggest that volumes derived from automated segmentation of T1-weighted structural images are reliable measures within the same scanner platform, even after upgrades; however, combining data across platform and across field-strength introduces a bias that should be considered in the design of multi-site studies, such as clinical drug trials. The results derived from the young groups (scanner upgrade effects and B1 inhomogeneity correction effects) should be considered as preliminary and in

  6. Integrated DNA and fingerprint analyses in the identification of 60-year-old mummified human remains discovered in an Alaskan glacier.

    PubMed

    Loreille, Odile M; Parr, Ryan L; McGregor, Kevin A; Fitzpatrick, Colleen M; Lyon, Chriss; Yang, Dongya Y; Speller, Camilla F; Grimm, Michael R; Grimm, Michael J; Irwin, Jodi A; Robinson, Edward M

    2010-05-01

    This report describes the identification of a merchant mariner who perished in 1948 when Northwest Airlines Flight 4422, a DC-4 carrying 24 seamen and six crew members crashed into Mount Sanford, Alaska. Fifty-one years later, a human forearm and hand were found close by the wreckage of the plane, prompting identification efforts using DNA and fingerprints. There were significant challenges to both the fingerprint and DNA analyses. The hand was badly desiccated, making fingerprint friction-ridge detail almost invisible and the remains had been embalmed upon discovery, making DNA amplification difficult. We present the results of an interdisciplinary approach that successfully addressed these challenges and ultimately led to the identification of the remains. These efforts relied on efficient fingerprint rejuvenation and imaging techniques that improved print resolution, as well as new DNA extraction techniques optimized for aggressively embalmed remains.

  7. Development of screening assays for nanoparticle toxicity assessment in human blood: preliminary studies with charged Au nanoparticles.

    PubMed

    Love, Sara A; Thompson, John W; Haynes, Christy L

    2012-09-01

    As nanoparticles have found increased use in both consumer and medical applications, corresponding increases in possible exposure to humans necessitate studies examining the impacts of these nanomaterials in biological systems. This article examines the effects of approximately 30-nm-diameter gold nanoparticles, with positively and negatively charged surface coatings in human blood. Here, we study the exposure effects, with up to 72 h of exposure to 5, 15, 25 and 50 µg/ml nanoparticles on hemolysis, reactive oxygen species (ROS) generation and platelet aggregation in subsets of cells from human blood. Assessing viability with hemolysis, results show significant changes in a concentration-dependent fashion. Rates of ROS generation were investigated using the dichlorofluorscein diacetate-based assay as ROS generation is a commonly suspected mechanism of nanoparticle toxicity; herein, ROS was not a significant factor. Optical monitoring of platelet aggregation revealed that none of the examined nanoparticles induced aggregation upon short-term exposure.

  8. Integrated network-diversity analyses suggest suppressive effect of Hodgkin’s lymphoma and slightly relieving effect of chemotherapy on human milk microbiome

    PubMed Central

    Ma, Zhanshan (Sam); Li, Lianwei; Li, Wendy; Li, Jie; Chen, Hongju

    2016-01-01

    We aim to investigate the effects of Hodgkin’s lymphoma and the chemotherapy for treating the disease on the human milk microbiome through integrated network and community diversity analyses. Our analyses suggest that Hodgkin’s lymphoma seems to have a suppressing effect on the milk microbiome by lowering the milk microbial community diversity, as measured by the Hill numbers profiles. Although the diversity analysis did not reveal an effect of chemotherapy on community diversity, bacterial species interaction network analysis shows that chemotherapy may help to slightly restore the milk microbiome impacted by Hodgkin’s lymphoma through its influence on the interactions among species (or OTUs). We further constructed diversity-metabolites network, which suggests that the milk microbial diversity is positively correlated with some beneficial milk metabolites such as DHA (DocosaHexaenoic Acid), and that the diversity is negatively correlated with some potentially harmful metabolites such as Butanal. We hence postulate that higher milk microbial diversity should be a signature of healthy mothers and beneficial to infants. Finally, we constructed metabolites OTU correlation networks, from which we identified some special OTUs. These OTUs deserve further investigations given their apparent involvements in regulating the levels of critical milk metabolites such as DHA, Inositol and Butanal. PMID:27386954

  9. Collecting Protein Biomarkers in Breath Using Electret Filters: A Preliminary Method on New Technical Model and Human Study.

    PubMed

    Li, Wang; Pi, Xitian; Qiao, Panpan; Liu, Hongying

    2016-01-01

    Biomarkers in exhaled breath are useful for respiratory disease diagnosis in human volunteers. Conventional methods that collect non-volatile biomarkers, however, necessitate an extensive dilution and sanitation processes that lowers collection efficiencies and convenience of use. Electret filter emerged in recent decade to collect virus biomarkers in exhaled breath given its simplicity and effectiveness. To investigate the capability of electret filters to collect protein biomarkers, a model that consists of an atomizer that produces protein aerosol and an electret filter that collects albumin and carcinoembryonic antigen-a typical biomarker in lung cancer development- from the atomizer is developed. A device using electret filter as the collecting medium is designed to collect human albumin from exhaled breath of 6 volunteers. Comparison of the collecting ability between the electret filter method and other 2 reported methods is finally performed based on the amounts of albumin collected from human exhaled breath. In conclusion, a decreasing collection efficiency ranging from 17.6% to 2.3% for atomized albumin aerosol and 42% to 12.5% for atomized carcinoembryonic antigen particles is found; moreover, an optimum volume of sampling human exhaled breath ranging from 100 L to 200 L is also observed; finally, the self-designed collecting device shows a significantly better performance in collecting albumin from human exhaled breath than the exhaled breath condensate method (p<0.05) but is not significantly more effective than reported 3-stage impactor method (p>0.05). In summary, electret filters are potential in collecting non-volatile biomarkers in human exhaled breath not only because it was simpler, cheaper and easier to use than traditional methods but also for its better collecting performance.

  10. Oxidation products of polyunsaturated fatty acids in infant formulas compared to human milk--a preliminary study.

    PubMed

    Michalski, Marie-Caroline; Calzada, Catherine; Makino, Asami; Michaud, Sabine; Guichardant, Michel

    2008-12-01

    Information about lipid oxidation in fresh and stored human milk compared with infant formulas is scarce. We aimed to assess n-6 and n-3 PUFA oxidation in these milks by measuring the 4-hydroxynonenal (4-HNE) and 4-hydroxyhexenal (4-HHE) content. Human milk samples (n = 4), obtained from volunteer mothers, were analyzed fresh and after 1 wk at 4 degrees C or 24 h at 18 degrees C. Vitamin E and malondialdehyde (MDA) were measured by HPLC and fatty acid profile by GC. The 4-HHE and 4-HNE contents were measured by GC-MS. Infant formulas (n = 10) were tested; their fat droplet size was measured by laser light scattering and observed by confocal laser scanning microscopy. Human milk samples contained 31.0 +/- 6.3 g/L of lipids and 1.14 +/- 0.26 mg/L of vitamin E. Fat droplets were smaller in infant formulas than reported in human milk. The (4-HHE/n-3 PUFA) ratio was 0.19 +/- 0.01 microg/g in fresh human milk (unchanged after storage) versus 3.6 +/- 3.1 microg/g in dissolved powder formulas and 4.3 +/- 3.8 microg/g in liquid formula. (4-HNE/n-6 PUFA) was 0.004 +/- 0.000 microg/g in fresh milk (0.03 +/- 0.01 microg/g after storage) versus 1.1 +/- 1.0 microg/g in dissolved powder formulas and 0.2 +/- 0.3 microg/g in liquid formula. Infant formulas also contained more MDA than human milk. n-3 PUFA were more prone to oxidation than n-6 PUFA. Whether threshold levels of 4-HHE and 4-HNE would be of health concern should be elucidated.

  11. Collecting Protein Biomarkers in Breath Using Electret Filters: A Preliminary Method on New Technical Model and Human Study

    PubMed Central

    Li, Wang; Pi, Xitian; Qiao, Panpan; Liu, Hongying

    2016-01-01

    Biomarkers in exhaled breath are useful for respiratory disease diagnosis in human volunteers. Conventional methods that collect non-volatile biomarkers, however, necessitate an extensive dilution and sanitation processes that lowers collection efficiencies and convenience of use. Electret filter emerged in recent decade to collect virus biomarkers in exhaled breath given its simplicity and effectiveness. To investigate the capability of electret filters to collect protein biomarkers, a model that consists of an atomizer that produces protein aerosol and an electret filter that collects albumin and carcinoembryonic antigen-a typical biomarker in lung cancer development- from the atomizer is developed. A device using electret filter as the collecting medium is designed to collect human albumin from exhaled breath of 6 volunteers. Comparison of the collecting ability between the electret filter method and other 2 reported methods is finally performed based on the amounts of albumin collected from human exhaled breath. In conclusion, a decreasing collection efficiency ranging from 17.6% to 2.3% for atomized albumin aerosol and 42% to 12.5% for atomized carcinoembryonic antigen particles is found; moreover, an optimum volume of sampling human exhaled breath ranging from 100 L to 200 L is also observed; finally, the self-designed collecting device shows a significantly better performance in collecting albumin from human exhaled breath than the exhaled breath condensate method (p<0.05) but is not significantly more effective than reported 3-stage impactor method (p>0.05). In summary, electret filters are potential in collecting non-volatile biomarkers in human exhaled breath not only because it was simpler, cheaper and easier to use than traditional methods but also for its better collecting performance. PMID:26934615

  12. PRELIMINARY FACTOR ANALYSIS OF VISUAL COGNITION AND MEMORY. STUDIES IN CINE-PSYCHOMETRY, FINAL REPORT, PART I.

    ERIC Educational Resources Information Center

    SEIBERT, WARREN F.; AND OTHERS

    PRELIMINARY ANALYSES WERE UNDERTAKEN TO DETERMINE THE POTENTIAL CONTRIBUTION OF MOTION PICTURE FILMS TO FACTOR ANALYTIC STUDIES OF HUMAN INTELLECT. OF PRIMARY CONCERN WERE THE OPERATIONS OF COGNITION AND MEMORY, FORMING TWO OF THE FIVE OPERATION COLUMNS OF GUILFORD'S "STRUCTURE OF INTELLECT." THE CORE REFERENCE FOR THE STUDY WAS DEFINED…

  13. Selected Bibliography on Culture and Cultural Materials, Preliminary Edition. Series A: Reference Materials, Human Relations in Cultural Context.

    ERIC Educational Resources Information Center

    Condon, E. C.; And Others

    Included in this bibliography are references to resources and materials available to the teacher and educator on human relations and cultural education. The bibliography is divided into three sections on culture, specific culture, and adult bilingual-bicultural education. The section on culture presents background information on the relation of…

  14. Purification, crystallization and preliminary X-ray characterization of a human mitochondrial phenylalanyl-tRNA synthetase

    SciTech Connect

    Levin, Inna; Kessler, Naama; Moor, Nina; Klipcan, Liron; Koc, Emine; Templeton, Paul; Spremulli, Linda; Safro, Mark

    2007-09-01

    The expression, purification and crystallization of recombinant human mitochondrial phenylalanyl-tRNA synthetase (mitPheRS) are reported. Diffraction data were collected to 2.2 Å resolution and the mitPheRS structure was solved using the molecular-replacement method. Human monomeric mitochondrial phenylalanyl-tRNA synthetase (mitPheRS) is an enzyme that catalyzes the charging of tRNA with the cognate amino acid phenylalanine. Human mitPheRS is a chimera of the bacterial α-subunit of PheRS and the B8 domain of its β-subunit. Together, the α-subunit and the ‘RNP-domain’ (B8 domain) at the C-terminus form the minimal structural set to construct an enzyme with phenylalanylation activity. The recombinant human mitPheRS was purified to homogeneity and crystallized in complex with phenylalanine and ATP. The crystals diffracted to 2.2 Å resolution and belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 55, b = 90, c = 96 Å.

  15. A preliminary human factors planning and design outline of parameters related to space station windows and CCTV monitoring

    NASA Technical Reports Server (NTRS)

    Haines, R. F.

    1985-01-01

    The question of the merits of placing windows on proposed future space stations is addressed. The use of windows for human visual capabilities is compared to using closed circuit television. Placement and field of view, as well as the number of windows is discussed.

  16. Preliminary study on the time-related changes of the infrared thermal images of the human body

    NASA Astrophysics Data System (ADS)

    Li, Ziru; Zhang, Xusheng; Lin, Gang; Chen, Zhigang

    2009-08-01

    It is of great importance to study the manifestations and the influencing factors of the time-related changes of infrared thermal images (ITI) of human body since the variable body surface temperature distribution seriously affected the application of ITI in medicine. In this paper, manifestations of time-related changes of the ITI of human body from three double-blind randomized trials and their correlation with meteorological factors (e.g. temperature, pressure, humidity, cold front passage and tropical cyclone landing) were studied. The trials were placebo or drug controlled studying the influences of Chinese medicine health food (including Shengsheng capsule with immunity adjustment function, Shengan capsule with sleep improvement function and Shengyi capsule with the function of helping to decrease serum lipid) on the ITI of human body. In the first thirty-six days of the trials images were scanned every six days and image data in the seven observation time spots (including the 0, 6, 12, 18, 24, 30, 36 day of the trial) were used for the time-related study. For every subject the scanned time was fixed in the day within two hours. The ITI features which could reflect the functions of the health foods were studied. The indexes of the features were relative magnitude (temperature difference between the viewing area and the reference area). Results showed that the variation tendencies of the trial group and control group were basically the same in placebo controlled trials and some of the long-term effects of Chinese medicine health food could be reflected significantly in certain time spots in the first thirty-six days. Time-related changes of the ITI of human body were closely related with meteorological factors but there were other influencing factors still need to be studied. As the ITI of human body could reflect the influences of Chinese medicine health foods and are closely related with meteorology, there are bright prospects for the application of ITI in

  17. Genome-wide analyses reveal a highly conserved Dengue virus envelope peptide which is critical for virus viability and antigenic in humans

    PubMed Central

    Fleith, Renata C.; Lobo, Francisco P.; dos Santos, Paula F.; Rocha, Mariana M.; Bordignon, Juliano; Strottmann, Daisy M.; Patricio, Daniel O.; Pavanelli, Wander R.; Lo Sarzi, Maria; Santos, Claudia N. D.; Ferguson, Brian J.; Mansur, Daniel S.

    2016-01-01

    Targeting regions of proteins that show a high degree of structural conservation has been proposed as a method of developing immunotherapies and vaccines that may bypass the wide genetic variability of RNA viruses. Despite several attempts, a vaccine that protects evenly against the four circulating Dengue virus (DV) serotypes remains elusive. To find critical conserved amino acids in dengue viruses, 120 complete genomes of each serotype were selected at random and used to calculate conservation scores for nucleotide and amino acid sequences. The identified peptide sequences were analysed for their structural conservation and localisation using crystallographic data. The longest, surface exposed, highly conserved peptide of Envelope protein was found to correspond to amino acid residues 250 to 270. Mutation of this peptide in DV1 was lethal, since no replication of the mutant virus was detected in human cells. Antibodies against this peptide were detected in DV naturally infected patients indicating its potential antigenicity. Hence, this study has identified a highly conserved, critical peptide in DV that is a target of antibodies in infected humans. PMID:27805018

  18. A three-dimensional model of the human blood-brain barrier to analyse the transport of nanoparticles and astrocyte/endothelial interactions

    PubMed Central

    Sreekanthreddy, Peddagangannagari; Gromnicova, Radka; Davies, Heather; Phillips, James; Romero, Ignacio A.; Male, David

    2016-01-01

    The aim of this study was to develop a three-dimensional (3D) model of the human blood-brain barrier in vitro, which mimics the cellular architecture of the CNS and could be used to analyse the delivery of nanoparticles to cells of the CNS. The model includes human astrocytes set in a collagen gel, which is overlaid by a monolayer of human brain endothelium (hCMEC/D3 cell line). The model was characterised by transmission electron microscopy (TEM), immunofluorescence microscopy and flow cytometry. A collagenase digestion method could recover the two cell types separately at 92-96% purity.  Astrocytes grown in the gel matrix do not divide and they have reduced expression of aquaporin-4 and the endothelin receptor, type B compared to two-dimensional cultures, but maintain their expression of glial fibrillary acidic protein. The effects of conditioned media from these astrocytes on the barrier phenotype of the endothelium was compared with media from astrocytes grown conventionally on a two-dimensional (2D) substratum. Both induce the expression of tight junction proteins zonula occludens-1 and claudin-5 in hCMEC/D3 cells, but there was no difference between the induced expression levels by the two media. The model has been used to assess the transport of glucose-coated 4nm gold nanoparticles and for leukocyte migration. TEM was used to trace and quantitate the movement of the nanoparticles across the endothelium and into the astrocytes. This blood-brain barrier model is very suitable for assessing delivery of nanoparticles and larger biomolecules to cells of the CNS, following transport across the endothelium. PMID:26870320

  19. Expression, purification, cocrystallization and preliminary crystallographic analysis of sucrose octasulfate/human complement regulator factor H SCRs 6–8

    SciTech Connect

    Prosser, Beverly E.; Johnson, Steven; Roversi, Pietro; Clark, Simon J.; Tarelli, Edward; Sim, Robert B.; Day, Antony J.; Lea, Susan M.

    2007-06-01

    The crystallization of human complement regulator FH-678{sub 402H} with a glycosaminoglycan analogue is described. Human plasma protein complement factor H (FH) is an inhibitor of the spontaneously activated alternative complement pathway. An allotypic variant of FH, 402His, has been associated with age-related macular degeneration, the leading cause of blindness in the elderly. Crystals of FH domains 6–8 (FH678) containing 402His have been grown in the presence of a polyanionic sucrose octasulfate ligand (an analogue of the natural glycosaminoglycan ligands of FH) using both native and selenomethionine-derivatized protein. Native data sets diffracting to 2.3 Å and SeMet data sets of up to 2.8 Å resolution have been collected. An anomalous difference Patterson map reveals self- and cross-peaks from two incorporated Se atoms. The corresponding selenium substructure has been solved.

  20. Crystallization and preliminary crystallographic studies of human kallikrein 7, a serine protease of the multigene kallikrein family

    PubMed Central

    Fernández, Israel S.; Ständker, Ludger; Forssmann, Wolf-Georg; Giménez-Gallego, Guillermo; Romero, Antonio

    2007-01-01

    Human kallikreins are a group of serine proteases of high sequence homology whose genes are grouped as a single cluster at chromosome 19. Although the physiological roles of kallikreins are generally still unknown, members of the kallikrein family have been clearly implicated in pathological situations such as cancer and psoriasis. Human kallikrein 7 (hK7) has been shown to be involved in pathological keratinization, psoriasis and ovarian cancer. In order to gain insight into the molecular structure of this protein, hK7 was crystallized after recombinant production in its folded and active form using a periplasmic secretion vector in Escherichia coli. The crystals belonged to the rhombohedral space group H32 and diffracted to 2.8 Å. The phase problem was solved by molecular replacement using the mouse kallikrein-related protein neuropsin. Completion of the model and structure refinement are under way. PMID:17671364

  1. Crystallization and preliminary crystallographic study of human coronavirus NL63 main protease in complex with an inhibitor.

    PubMed

    Wang, Fenghua; Tan, Yusheng; Li, Huiyan; Chen, Xia; Wang, Jinshan; Li, Shuang; Fu, Sheng; Zhao, Qi; Chen, Cheng; Su, Dan; Yang, Haitao

    2014-08-01

    Human coronavirus NL63 mainly infects younger children and causes cough, fever, rhinorrhoea, bronchiolitis and croup. It encodes two polyprotein precursors required for genome replication and transcription. Each polyprotein undergoes extensive proteolytic processing, resulting in functional subunits. This process is mainly mediated by its genome-encoded main protease, which is an attractive target for antiviral drug design. In this study, the main protease of human coronavirus NL63 was crystallized in complex with a Michael acceptor. The complex crystals diffracted to 2.85 Å resolution and belonged to space group P41212, with unit-cell parameters a = b = 87.2, c = 212.1 Å. Two molecules were identified per asymmetric unit.

  2. A preliminary analysis of human factors affecting the recognition accuracy of a discrete word recognizer for C3 systems

    NASA Astrophysics Data System (ADS)

    Yellen, H. W.

    1983-03-01

    Literature pertaining to Voice Recognition abounds with information relevant to the assessment of transitory speech recognition devices. In the past, engineering requirements have dictated the path this technology followed. But, other factors do exist that influence recognition accuracy. This thesis explores the impact of Human Factors on the successful recognition of speech, principally addressing the differences or variability among users. A Threshold Technology T-600 was used for a 100 utterance vocubalary to test 44 subjects. A statistical analysis was conducted on 5 generic categories of Human Factors: Occupational, Operational, Psychological, Physiological and Personal. How the equipment is trained and the experience level of the speaker were found to be key characteristics influencing recognition accuracy. To a lesser extent computer experience, time or week, accent, vital capacity and rate of air flow, speaker cooperativeness and anxiety were found to affect overall error rates.

  3. Crystallization and preliminary crystallographic analysis of human Ca{sup 2+}-loaded calbindin-D28k

    SciTech Connect

    Zhang, Chang; Sun, Yuna; Wang, Wei; Zhang, Yan; Ma, Ming; Lou, Zhiyong

    2008-02-01

    Human calbindin-D28k has been overexpressed in E. coli, purified and crystallized. Diffraction data were collected to 2.4 Å resolution. Calbindin-D28k is a calcium-binding protein that belongs to the troponin C superfamily. It is expressed in many tissues, including brain, intestine, kidney and pancreas, and performs roles as both a calcium buffer and a calcium sensor and carries out diverse physiological functions of importance. In order to resolve the crystal structure of human calbindin-D28k and to gain a better understanding of its biological functions, recombinant human calbindin-D28k was crystallized at 291 K using PEG 3350 as precipitant and a 2.4 Å resolution X-ray data set was collected from a single flash-cooled crystal (100 K). The crystal belonged to space group C2, with unit-cell parameters a = 108.1, b = 28.2, c = 70.6 Å, β = 107.8°. The presence of one molecule per asymmetric unit is presumed, corresponding to a Matthews coefficient of 1.75 Å{sup 3} Da{sup −1}.

  4. Crystallization and preliminary crystallographic studies of human kallikrein 7, a serine protease of the multigene kallikrein family

    SciTech Connect

    Fernández, Israel S.; Ständker, Ludger; Forssmann, Wolf-Georg; Giménez-Gallego, Guillermo; Romero, Antonio

    2007-08-01

    The cloning, expression, purification and crystallization of recombinant human kallikrein 7, directly synthesized in the active form in E. coli, is described. Diffraction data were collected to 2.8 Å resolution from native crystals. Human kallikreins are a group of serine proteases of high sequence homology whose genes are grouped as a single cluster at chromosome 19. Although the physiological roles of kallikreins are generally still unknown, members of the kallikrein family have been clearly implicated in pathological situations such as cancer and psoriasis. Human kallikrein 7 (hK7) has been shown to be involved in pathological keratinization, psoriasis and ovarian cancer. In order to gain insight into the molecular structure of this protein, hK7 was crystallized after recombinant production in its folded and active form using a periplasmic secretion vector in Escherichia coli. The crystals belonged to the rhombohedral space group H32 and diffracted to 2.8 Å. The phase problem was solved by molecular replacement using the mouse kallikrein-related protein neuropsin. Completion of the model and structure refinement are under way.

  5. Preliminary study on the relationship between cAMP level and gsp expression in cultured human pituitary somatotrophinomas.

    PubMed

    Lei, T; Liu, Q; Li, L; Zhang, L; Shu, K; Xue, D

    2000-01-01

    In order to investigate the relationship between abnormal intracellular signal transduction and tumorgenesis of human pituitary somatotrophinomas, the effects of protein kinase A (PKA)-dependent growth hormone (GH) releasing hormone (GHRH) and protein kinase C (PKC)-dependent GH-releasing peptide (GHRP-6) on cAMP production were observed by using cell culture and biochemical methods, and the expression of the gsp oncogene was detected by using PCR and direct sequence assay methods in 11 patients with human pituitary somatotrophinomas. It was found that GHRP-6 exerted significant stimulatory effect on cAMP production by 2 gsp-positive tumors and no effect on the gsp-negative tumors. GHRP-6 could enhance the stimulation of cAMP production induced by GHRH in tumor without gsp oncogenes. It was suggested that both GHRH and GHRP-6 exert identical effects on human pituitary soamtotrophinomas, which was contributed to the cross-talk between the two intracellular signal transduction pathways in pituitary cells.

  6. Overproduction and preliminary crystallographic study of a human kynurenine aminotransferase II homologue from Pyrococcus horikoshii OT3

    SciTech Connect

    Chon, Hyongi; Matsumura, Hiroyoshi; Shimizu, Shoko; Maeda, Nao; Koga, Yuichi; Takano, Kazufumi; Kanaya, Shigenori

    2005-03-01

    A human kynurenine aminotransferase II homologue from P. horikoshii OT3 has been overproduced in E. coli, purified, and characterized. Crystals of this protein have been obtained and analyzed by X-ray diffraction. The Pyrococcus horikoshii OT3 genome contains a gene encoding a human kynurenine aminotransferase II (KAT II) homologue, which consists of 428 amino-acid residues and shows an amino-acid sequence identity of 30% to human KAT II. This gene was overexpressed in Escherichia coli and the recombinant protein (Ph-KAT II) was purified. Gel-filtration chromatography showed that Ph-KAT II exists as a homodimer. Ph-KAT II exhibited enzymatic activity that catalyzes the transamination of l-kynurenine to produce kynurenic acid. Crystals of Ph-KAT II were grown using the sitting-drop vapour-diffusion method and native X-ray diffraction data were collected to 2.2 Å resolution using synchrotron radiation from station BL44XU at SPring-8. The crystals belong to the centred orthorhombic space group C222{sub 1}, with unit-cell parameters a = 71.75, b = 86.84, c = 137.30 Å. Assuming one molecule per asymmetric unit, the V{sub M} value was 2.19 Å{sup 3} Da{sup −1} and the solvent content was 43.3%.

  7. [Human activities, hydro-agricultural management and urinary schistosomiasis. Methodological approach and results (a preliminary study in Upper-Volta)].

    PubMed

    Le Bras, M; Faucher, P; Giap, G; Meric, D; Commenges, D; Villenave, D; Camara, S; Gatheron, C; Appriou, M

    1982-01-01

    In the schistosomiasis endemic sub-sahelian region, a study of the prevalence of the disease was carried out on three different areas: the highest was around a small river, the lowest in the hydro-agricultural management region, mean prevalence was noted around a natural lake. Human activities have been classified with regards to the areas of transmission. Activity is closely linked to prevalence. For the populations, symptoms of urinary schistosomiasis (Rudzenga) constitute a real disease, traditional healers are the masters of earth (Tengsoba), water has hardly any importance in transmission.

  8. Supplementary report on antilock analyses

    NASA Technical Reports Server (NTRS)

    Zellner, J. W.

    1985-01-01

    Generic modulator analysis was performed to quantify the effects of dump and reapply pressure rates on antilock stability and performance. Analysis will include dump and reapply rates, and lumped modulator delay. Based on the results of the generic modulator analysis and earlier toggle optimization analysis (with Mitsubishi modulator), a recommended preliminary antilock design was synthesized and its response and performance simulated. The results of these analyses are documented.

  9. IS1311 and IS1245 Restriction Fragment Length Polymorphism Analyses, Serotypes, and Drug Susceptibilities of Mycobacterium avium Complex Isolates Obtained from a Human Immunodeficiency Virus-Negative Patient

    PubMed Central

    Dvorska, Lenka; Bartos, Milan; Ostadal, Oldrich; Kaustova, Jarmila; Matlova, Ludmila; Pavlik, Ivo

    2002-01-01

    Six isolates of Mycobacterium avium of genotype dnaJ+ IS901− IS1311+ IS1245+ and serotypes 6 (n = 1), 6/9, (n = 2), and 9 (n = 3) were obtained within a 5-month period from a human immunodeficiency virus-negative patient treated for tuberculosis. The isolates were identified with PvuII restriction fragment length polymorphism (RFLP) analysis as a single IS1311 RFLP type and six different IS1245 RFLP types. Six separate colonies/clones obtained by subculture from each of the six isolates were tested for MICs of a set of 10 drugs. This report documents the appearance of isolates that are resistant to antimycobacterial drugs as the duration of therapy increases. Because isolates recovered from the patient following longer duration of treatment were more likely to be resistant to more antimycobacterial drugs, we would conclude that there was selection for antimycobacterial drug-resistant isolates. Analyses of all 36 clones identified three IS1311 and 22 IS1245 types forming three clusters. Tests of 105 environmental samples collected in the home and the work place of the patient yielded 16 mycobacterial isolates, of which one M. avium from soil was of genotype dnaJ+ IS901+ IS1311+ IS1245+ and serotype 2, and the second M. avium from a vacuum cleaner was of genotype dnaJ+ IS901− IS1311+ IS1245+ and serotype 9. Overall analyses of the results did not reveal any relation between serotype, RFLP type, and drug susceptibility. Based on the course of the disease in the patient and different serotypes, IS1311 and IS1245 RFLP types of isolates of M. avium we suppose represent polyclonal infection. PMID:12354870

  10. Advances in Thin Tissue Golgi-Cox Impregnation: Fast, Reliable Methods for Multi-Assay Analyses in Rodent and Non-human Primate Brain

    PubMed Central

    Levine, Nathan D.; Rademacher, David J.; Collier, Timothy J.; O’Malley, Jennifer A.; Kells, Adrian P.; Sebastian, Waldy San; Bankiewicz, Krystof S.; Steece-Collier, Kathy

    2013-01-01

    In 1873 Camillo Golgi discovered a staining technique that allowed for the visualization of whole neurons within the brain, initially termed ‘the black reaction’ and is now known as Golgi impregnation. Despite the capricious nature of this method, Golgi impregnation remains a widely used method for whole neuron visualization and analysis of dendritic arborization and spine quantification. We describe a series of reliable, modified ‘Golgi-Cox’ impregnation methods that complement some existing methods and have several advantages over traditional whole brain ‘Golgi’ impregnation. First, these methods utilize 60–100μm thick brain sections, which allows for fast, reliable impregnation of neurons in rats (7–14 days) and non-human primates (NHP) (30 days) while avoiding the pitfalls of other ‘rapid Golgi’ techniques traditionally employed with thin sections. Second, these methods employ several common tissue fixatives, resulting in high quality neuron impregnation in brain sections from acrolein, glutaraldehyde, and paraformaldehyde perfused rats, and in glutaraldehyde perfused NHP brain tissue. Third, because thin sections are obtained on a vibratome prior to processing, alternate sections of brain tissue can be used for additional analyses such as immunohistochemistry or electron microscopy. This later advantage allows for comparison of, for example, dendrite morphology in sections adjacent to pertinent histochemical markers or ultrastructural components. Finally, we describe a method for simultaneous light microscopic visualization of both tyrosine hydroxylase immunohistochemistry and Golgi impregnation in the same tissue section. Thus, the methods described here allow for fast, high quality Golgi impregnation and conserve experimental subjects by allowing multiple analyses within an individual animal. PMID:23313849

  11. 17β-estradiol effects on human coronaries and grafts employed in myocardial revascularization: a preliminary study

    PubMed Central

    Polvani, Gianluca; Barili, Fabio; Rossoni, Giuseppe; Dainese, Luca; Ossola, Manuela Wally; Topkara, Veli K; Grillo, Francesco; Penza, Eleonora; Tremoli, Elena; Biglioli, Paolo

    2006-01-01

    Background This study was undertaken to compare the in vitro effects of 17β-estradiol on human epicardial coronary arteries, resistance coronary arteries and on arterial vessels usually employed as grafts in surgical myocardial revascularization. Methods Coronary artery rings (descending coronary artery, right coronary artery, circumflex coronary artery, first septal branch) and arterial graft rings (internal thoracic artery, gastro-epiploic artery) obtained from human heart donors with heart not suitable to cardiac transplantation were connected to force transducer for isometric force recording. Precontracted specimens with and without endothelium were exposed to increasing concentration of 17β-estradiol (3–30–300–3000 nmol/l) and to vehicle (0.1% v/v ethanol). We also evaluated the effects of 17β-estradiol on vessels before and 20 minutes after exposure to L-monomethyl-arginine and indomethacin. Results 17β-estradiol induced a significant relaxation in all precontracted vessels (mean maximum effect: 78,6% ± 8,5). This effect was not different among the different rings and was not related to the presence of endothelium. N-monomethyl-L-arginine and indomethacin did not modify 17β-estradiol relaxant effect. Conclusion The vasodilator action of the 17β-estradiol is similar on coronary arteries, resistance coronary arteries and arterial vessels usually employed as grafts in myocardial revascularization. PMID:17181858

  12. A preliminary study of viral metagenomics of French bat species in contact with humans: identification of new mammalian viruses.

    PubMed

    Dacheux, Laurent; Cervantes-Gonzalez, Minerva; Guigon, Ghislaine; Thiberge, Jean-Michel; Vandenbogaert, Mathias; Maufrais, Corinne; Caro, Valérie; Bourhy, Hervé

    2014-01-01

    The prediction of viral zoonosis epidemics has become a major public health issue. A profound understanding of the viral population in key animal species acting as reservoirs represents an important step towards this goal. Bats harbor diverse viruses, some of which are of particular interest because they cause severe human diseases. However, little is known about the diversity of the global population of viruses found in bats (virome). We determined the viral diversity of five different French insectivorous bat species (nine specimens in total) in close contact with humans. Sequence-independent amplification, high-throughput sequencing with Illumina technology and a dedicated bioinformatics analysis pipeline were used on pooled tissues (brain, liver and lungs). Comparisons of the sequences of contigs and unassembled reads provided a global taxonomic distribution of virus-related sequences for each sample, highlighting differences both within and between bat species. Many viral families were present in these viromes, including viruses known to infect bacteria, plants/fungi, insects or vertebrates, the most relevant being those infecting mammals (Retroviridae, Herpesviridae, Bunyaviridae, Poxviridae, Flaviviridae, Reoviridae, Bornaviridae, Picobirnaviridae). In particular, we detected several new mammalian viruses, including rotaviruses, gammaretroviruses, bornaviruses and bunyaviruses with the identification of the first bat nairovirus. These observations demonstrate that bats naturally harbor viruses from many different families, most of which infect mammals. They may therefore constitute a major reservoir of viral diversity that should be analyzed carefully, to determine the role played by bats in the spread of zoonotic viral infections.

  13. Recombinant expression, purification and preliminary biophysical and structural studies of C-terminal carbohydrate recognition domain from human galectin-4.

    PubMed

    Rustiguel, Joane K; Kumagai, Patricia S; Dias-Baruffi, Marcelo; Costa-Filho, Antonio J; Nonato, Maria Cristina

    2016-02-01

    Galectin-4 (Gal4), a tandem-repeat type galectin, is expressed in healthy epithelium of the gastrointestinal tract. Altered levels of Gal4 expression are associated with different types of cancer, suggesting its usage as a diagnostic marker as well as target for drug development. The functional data available for this class of proteins suggest that the wide spectrum of cellular activities reported for Gal4 relies on distinct glycan specificity and structural characteristics of its two carbohydrate recognition domains. In the present work, two independent constructs for recombinant expression of the C-terminal domain of human galectin-4 (hGal4-CRD2) were developed. His6-tagged and untagged recombinant proteins were overexpressed in Escherichia coli, and purified by affinity chromatography followed by gel filtration. Correct folding and activity of hGal4-CRD2 were assessed by circular dichroism and fluorescence spectroscopies, respectively. Diffraction quality crystals were obtained by vapor-diffusion sitting drop setup and the crystal structure of CRD2 was solved by molecular replacement techniques at 1.78 Å resolution. Our work describes the development of important experimental tools that will allow further studies in order to correlate structure and binding properties of hGal4-CRD2 and human galectin-4 functional activities.

  14. Crystallization and preliminary X-ray diffraction data for the aconitase form of human iron-regulatory protein 1

    SciTech Connect

    Dupuy, J.; Darnault, C.; Moulis, J. M.

    2005-05-01

    Two crystal forms of the aconitase version of recombinant human IRP1 are reported. Iron-regulatory proteins (IRPs) 1 and 2 are closely related molecules involved in animal iron metabolism. Both proteins can bind to specific mRNA regions called iron-responsive elements and thereby control the expression of proteins involved in the uptake, storage and utilization of iron. In iron-replete cells, IRP1, but not IRP2, binds a [4Fe–4S] cluster and functions as a cytoplasmic aconitase, with simultaneous loss of its RNA-binding ability. Whereas IRP2 is known to be involved in Fe homeostasis, the role of IRP1 is less clear; it may provide a link between citrate and iron metabolisms and be involved in oxidative stress response. Here, two crystal forms of the aconitase version of recombinant human IRP1 are reported. An X-ray fluorescence measurement performed on a gold-derivative crystal showed the unexpected presence of zinc, in addition to gold and iron. Both native and MAD X-ray data at the Au, Fe and Zn absorption edges have been collected from these crystals.

  15. Experimental analysis of the mechanical behavior of the viscoelastic porcine pancreas and preliminary case study on the human pancreas.

    PubMed

    Wex, C; Fröhlich, M; Brandstädter, K; Bruns, C; Stoll, A

    2015-01-01

    The aim of this article is to study the mechanical properties of the pancreas. Up to now, the mechanical properties of the pancreas are not sufficiently characterized. The possibility of intraoperative mechanical testing of pathological pancreata will allow the classification of pancreatic diseases in the future. The application of mechanical parameters instead of the intraoperative frozen section analysis shortens waiting times in the operating room. This study proves the general applicability of shear rheology for the determination of the mechanical properties of pancreas and the assessment of graft quality for transplantation. Porcine and human pancreas samples were examined ex vivo and a nonlinear viscoelastic behavior was observed. Pancreas was found to be more viscous than liver but both abdominal organs showed a similar flow behavior. The shear deformation dependence of healthy human pancreas was similar to porcine pancreas. An increase in the post-mortem time led to an increase in the complex modulus for a post-mortem time up to 8.5 days. Histological investigations showed that an increased amount of collagen coincides with the stiffening of the pancreatic tissue.

  16. Purification, crystallization and preliminary crystallographic analysis of the globular domain of the human type V myosin Myo5a.

    PubMed

    Velvarska, Hana; Niessing, Dierk

    2013-11-01

    Type V myosins constitute the main cargo-transporting class of myosin motors in higher eukaryotes. They are mainly defined by their C-terminal globular domain, which is required for cargo binding as well as for motor auto-inhibition in the absence of cargo. To date, high-resolution structures only exist for globular domains from yeast. Since the majority of cellular cargoes in yeast are very different from the cargoes in higher eukaryotes, structural insights into the domain organization of globular domains from human type V myosins are important. The globular domain of human Myo5a was cloned, expressed and crystallized and data sets were collected. The crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 75.04, b = 86.70, c = 131.41 Å, α = β = γ = 90°, and diffracted with data-collection quality to 2.5 Å resolution.

  17. Syzygium cumini (Jamun) reduces the radiation-induced DNA damage in the cultured human peripheral blood lymphocytes: a preliminary study.

    PubMed

    Jagetia, Ganesh Chandra; Baliga, Manjeshwar Shrinath

    2002-06-07

    The effects of various concentrations (0.0, 1.56, 3.125, 6.25, 12.5, 25, 50 and 100 microg/ml) of the leaf extract of Syzygium cumini Linn. or Eugenia cumini (SC; black plum, Jamun, family Myrtaceae) was studied on the alteration in the radiation-induced micronuclei formation in the cultured human peripheral blood lymphocytes. Treatment of lymphocytes to various concentrations of SC resulted in a dose dependent increase in the micronuclei-induction, especially after 25-100 microg/ml extract. The exposure of human lymphocytes to various concentrations of SC extract before 3 Gy gamma-irradiation resulted in a significant decline in the micronuclei-induction at all the drug doses when compared with the non-drug treated irradiated cultures. A nadir in MNBNC frequency was observed for 12.5 microg/ml drug concentration, where the MNBNC frequency was approximately fourfold lower than that of the non-drug treated irradiated cultures. Therefore, this dose may be considered as an optimum dose for radiation protection. Our study demonstrates that the leaf extract of S. cumini, a plant traditionally used to treat diabetic disorders protects against the radiation-induced DNA damage.

  18. A Preliminary Study of Viral Metagenomics of French Bat Species in Contact with Humans: Identification of New Mammalian Viruses

    PubMed Central

    Dacheux, Laurent; Cervantes-Gonzalez, Minerva; Guigon, Ghislaine; Thiberge, Jean-Michel; Vandenbogaert, Mathias; Maufrais, Corinne

    2014-01-01

    The prediction of viral zoonosis epidemics has become a major public health issue. A profound understanding of the viral population in key animal species acting as reservoirs represents an important step towards this goal. Bats harbor diverse viruses, some of which are of particular interest because they cause severe human diseases. However, little is known about the diversity of the global population of viruses found in bats (virome). We determined the viral diversity of five different French insectivorous bat species (nine specimens in total) in close contact with humans. Sequence-independent amplification, high-throughput sequencing with Illumina technology and a dedicated bioinformatics analysis pipeline were used on pooled tissues (brain, liver and lungs). Comparisons of the sequences of contigs and unassembled reads provided a global taxonomic distribution of virus-related sequences for each sample, highlighting differences both within and between bat species. Many viral families were present in these viromes, including viruses known to infect bacteria, plants/fungi, insects or vertebrates, the most relevant being those infecting mammals (Retroviridae, Herpesviridae, Bunyaviridae, Poxviridae, Flaviviridae, Reoviridae, Bornaviridae, Picobirnaviridae). In particular, we detected several new mammalian viruses, including rotaviruses, gammaretroviruses, bornaviruses and bunyaviruses with the identification of the first bat nairovirus. These observations demonstrate that bats naturally harbor viruses from many different families, most of which infect mammals. They may therefore constitute a major reservoir of viral diversity that should be analyzed carefully, to determine the role played by bats in the spread of zoonotic viral infections. PMID:24489870

  19. A Preliminary Investigation into the Impact of a Pesticide Combination on Human Neuronal and Glial Cell Lines In Vitro

    PubMed Central

    Coleman, Michael D.; O'Neil, John D.; Woehrling, Elizabeth K.; Ndunge, Oscar Bate Akide; Hill, Eric J.; Menache, Andre; Reiss, Claude J.

    2012-01-01

    Many pesticides are used increasingly in combinations during crop protection and their stability ensures the presence of such combinations in foodstuffs. The effects of three fungicides, pyrimethanil, cyprodinil and fludioxonil, were investigated together and separately on U251 and SH-SY5Y cells, which can be representative of human CNS glial and neuronal cells respectively. Over 48h, all three agents showed significant reductions in cellular ATP, at concentrations that were more than tenfold lower than those which significantly impaired cellular viability. The effects on energy metabolism were reflected in their marked toxic effects on mitochondrial membrane potential. In addition, evidence of oxidative stress was seen in terms of a fall in cellular thiols coupled with increases in the expression of enzymes associated with reactive species formation, such as GSH peroxidase and superoxide dismutase. The glial cell line showed significant responsiveness to the toxin challenge in terms of changes in antioxidant gene expression, although the neuronal SH-SY5Y line exhibited greater vulnerability to toxicity, which was reflected in significant increases in caspase-3 expression, which is indicative of the initiation of apoptosis. Cyprodinil was the most toxic agent individually, although oxidative stress-related enzyme gene expression increases appeared to demonstrate some degree of synergy in the presence of the combination of agents. This report suggests that the impact of some pesticides, both individually and in combinations, merits further study in terms of their impact on human cellular health. PMID:22880100

  20. The Difference between Growth Factor Expression after Single and Multiple Fractures: Preliminary Results in Human Fracture Healing

    PubMed Central

    Binder, Harald; Eipeldauer, Stefan; Gregori, Markus; Höchtl-Lee, Leonard; Thomas, Anita; Tiefenboeck, Thomas M.; Hajdu, Stefan; Sarahrudi, Kambiz

    2015-01-01

    Objectives. Circulating levels of VEGF-A (Vascular Endothelia Growth Factor-A), TGF-β1 (Transforming Growth Factor-beta 1), and M-CSF (Macrophage-Colony Stimulating Factor) were found to be predictors of bone healing and therefore prognostic criteria of delayed bone healing or nonunion. The aim of this study was to evaluate a potential rise of these markers in patients with multiple fractures of long bones compared to patients with single fractured long bone. Methods. 92 patients were included in the study and finally after excluding all female patients 45 male patients were left for final analysis and divided into the single or multiple fracture group. TGF-β1, M-CSF, and VEGF-A serum levels were analysed over a time period of two weeks. Results. MCSF serum concentrations were higher in the group with multiple fractures as also TGF-β1 serum concentrations were at one and two weeks after trauma. No statistically significant difference was observed in the VEGF-A serum concentrations of both groups at either measurement point. Conclusion. We did observe a correlation between the quantity of the M-CSF and TGF-β1 expressions in serum and the number of fractured bones; surprisingly there was no statistically significant difference in the serum levels between patients with single and multiple fractures of long bones. PMID:26246654

  1. Identification and evolutionary dynamics of two novel human coronavirus OC43 genotypes associated with acute respiratory infections: phylogenetic, spatiotemporal and transmission network analyses

    PubMed Central

    Oong, Xiang Yong; Ng, Kim Tien; Takebe, Yutaka; Ng, Liang Jie; Chan, Kok Gan; Chook, Jack Bee; Kamarulzaman, Adeeba; Tee, Kok Keng

    2017-01-01

    Human coronavirus OC43 (HCoV-OC43) is commonly associated with respiratory tract infections in humans, with five genetically distinct genotypes (A to E) described so far. In this study, we obtained the full-length genomes of HCoV-OC43 strains from two previously unrecognized lineages identified among patients presenting with severe upper respiratory tract symptoms in a cross-sectional molecular surveillance study in Kuala Lumpur, Malaysia, between 2012 and 2013. Phylogenetic, recombination and comparative genomic analyses revealed two distinct clusters diverging from a genotype D-like common ancestor through recombination with a putative genotype A-like lineage in the non-structural protein (nsp) 10 gene. Signature amino acid substitutions and a glycine residue insertion at the N-terminal domain of the S1 subunit of the spike gene, among others, exhibited further distinction in a recombination pattern, to which these clusters were classified as genotypes F and G. The phylogeographic mapping of the global spike gene indicated that the genetically similar HCoV-OC43 genotypes F and G strains were potentially circulating in China, Japan, Thailand and Europe as early as the late 2000s. The transmission network construction based on the TN93 pairwise genetic distance revealed the emergence and persistence of multiple sub-epidemic clusters of the highly prevalent genotype D and its descendant genotypes F and G, which contributed to the spread of HCoV-OC43 in the region. Finally, a more consistent nomenclature system for non-recombinant and recombinant HCoV-OC43 lineages is proposed, taking into account genetic recombination as an important feature in HCoV evolution and classification. PMID:28050020

  2. Crystallization and Preliminary X-ray analysis of Human Recombinant Acid beta-glucocerebrosidase, a treatment for Gaucher's Disease

    NASA Technical Reports Server (NTRS)

    Roeber, Dana F.; Achari, Aniruddha; Manavalan, Partha; Edmunds, Tim; Scott, David L.; Curreri, Peter A. (Technical Monitor)

    2002-01-01

    Acid beta-glucocerebrosidase (N-acylsphingosyl - O - beta-D - glucoside:glucohydrolase) is a lysosomal glycoprotein that catalyzes the hydrolysis of the glycolipid glucocerebroside to glucose and ceramide. Inadequate levels of this enzyme underly the pathophysiology of Gaucher's disease. Cerezyme(R) (Genzyme Corporation, Cambridge, MA) is a partially deglycosylated form of recombinant human acid beta-glucocerebrosidase that is commercially available for the treatment of Gaucher patients. Although acid beta-glucocerebrosidase belongs to a large family of glycosidases, relatively little is known regarding its structural biology. We report the crystallization and the initial diffraction analysis of Cerezyme(R). The crystals are C-centered orthorhombic, with unit-cell parameters of a = 285.0 A, b = 110.2 A, and c = 91.7 A. A 99.9 A complete data set has been collected to 2.75 A with an R(sub sym) of 8.8 %.

  3. Human T-cell lymphotropic virus type III infection of the central nervous system: a preliminary in situ analysis

    SciTech Connect

    Stoler, M.H.; Eskin, T.A.; Benn, S.; Angerer, R.C.; Angerer, L.M.

    1986-11-07

    Patients with acquired immunodeficiency syndrome (AIDS) are subject to a spectrum of central nervous system (CNS) disorders. Recent evidence implicates the human T-cell lymphotropic virus type III (HTLV-III) in the pathogenesis of some of these illnesses, although the cells infected by the virus have yet to be identified. Using in situ hybridization, the authors examined brain tissue from two patients with AIDS encephalopathy for the presence of HTLV-III RNA. In both cases, viral RNA was detected and concentrated in, though not limited to, the white matter. The CNS cells most frequently infected included macrophages, pleomorphic microglia, and multinucleated giant cells. Less frequently, cells morphologically consistent with astrocytes, oligodendroglia, and rarely neurons were also infected. The findings strengthen the association of HTLV-III with the pathogenesis of AIDS encephalopathy. In situ hybridization can be applied to routinely prepared biopsy tissue in the diagnosis of HTLV-III infection of the CNS.

  4. Preliminary quantitative microradiography study into the distribution of bone mineralization within the basal bone of the human edentulous mandible.

    PubMed

    Hobson, R S; Beynon, A D

    1997-07-01

    Six edentulous human hemimandibles (three male, three female) of similar ages were sectioned transversely at the midline, mental foramen, midway between mental foramen and angle (body), and angle of the mandible. Planoparallel sections were prepared of the cortical plate on buccal and lingual surfaces and microradiographed alongside an aluminium step-wedge for computerized quantitative microradiography. Mean mineralization values and mineralization frequency distribution curves were calculated. Gender had no effect on the mean mineralization values, but did show a significant difference in the mineralization frequency distribution. There was no difference in mean mineralization between lingual and buccal cortical plates, but the distribution curves differed, with the lingual cortex distributions being more uniform. Each sample site was significantly different from all others, with the mental foramen and body sites showing the greatest variation in distribution of mineralization level.

  5. The preliminary solution structure of human p8MTCP1, a protein encoded by the putative MTCP1 oncogene

    NASA Astrophysics Data System (ADS)

    Barthe, P.; Guignard, L.; Yang, Y.-S.; Chiche, L.; Strub, M.-P.; Hoh, F.; Stern, M.-H.; Roumestand, C.

    1998-02-01

    The NMR solution structure of the cystein-rich human p8 protein coded by the oncogene MTCP1 reveals an original scaffold consisting of three α-helices, two of them tightly held together by two disulphide bridges in an antiparallel α-hairpin. MTCP1 was found to be expressed in mature prolymphocytic leukemias. La structure RMN de la protéine humaine p8 en solution présente un repliement original en trois hélices α, deux d'entre elles étant étroitement maintenues dans une orientation antiparallèle par deux ponts disulfures. Cette protéine riche en cystéine est codée par l'oncogène MTCP1 qui est exprimé dans des leucémies prolymphocytaires de phénotype mature.

  6. Using Virtual Technology to Promote Functional Communication in Aphasia: Preliminary Evidence From Interactive Dialogues With Human and Virtual Clinicians

    PubMed Central

    Martin, Nadine; Keshner, Emily; Rudnicky, Alex; Shi, Justin; Teodoro, Gregory

    2015-01-01

    Purpose We investigated the feasibility of using a virtual clinician (VC) to promote functional communication abilities of persons with aphasia (PWAs). We aimed to determine whether the quantity and quality of verbal output in dialogues with a VC would be the same or greater than those with a human clinician (HC). Method Four PWAs practiced dialogues for 2 sessions each with a HC and VC. Dialogues from before and after practice were transcribed and analyzed for content. We compared measures taken before and after practice in the VC and HC conditions. Results Results were mixed. Participants either produced more verbal output with the VC or showed no difference on this measure between the VC and HC conditions. Participants also showed some improvement in postpractice narratives. Conclusion Results provide support for the feasibility and applicability of virtual technology to real-life communication contexts to improve functional communication in PWAs. PMID:26431390

  7. Crystallization and preliminary X-ray analysis of recombinant human acid beta-glucocerebrosidase, a treatment for Gaucher's disease

    NASA Technical Reports Server (NTRS)

    Roeber, Dana; Achari, Aniruddha; Manavalan, Partha; Edmunds, Tim; Scott, David L.

    2003-01-01

    Acid beta-glucocerebrosidase (N-acylsphingosyl-1-O-beta-D-glucoside:glucohydrolase) is a lysosomal glycoprotein that catalyzes the hydrolysis of the glycolipid glucocerebroside to glucose and ceramide. Inadequate levels of this enzyme underly the pathophysiology of Gaucher's disease. Cerezyme (Genzyme Corporation, Cambridge, MA, USA) is a partially deglycosylated form of recombinant human acid beta-glucocerebrosidase that is used in the treatment of Gaucher patients. Although acid beta-glucocerebrosidase belongs to a large family of glycosidases, relatively little is known regarding its structural biology. Here, the crystallization and the initial diffraction analysis of Cerezyme are reported. The crystals are C-centered orthorhombic, with unit-cell parameters a = 285.0, b = 110.2, c = 91.7 A. A 99.9% complete data set has been collected to 2.75 A with an R(sym) of 8.8%.

  8. Hair as an indicator of the body content of polonium in humans: preliminary results from study of five male volunteers.

    PubMed

    Rääf, C L; Holstein, H; Holm, E; Roos, P

    2015-03-01

    The radionuclide (210)Po is of importance from a radiation protection view and has properties that cause special problems when attempting to determine the body content in humans. Estimates have traditionally been made from either urine and/or fecal samples, which require a time-consuming radiochemical preparation before alpha spectrometric determination. In order to find a more simple and less labor intensive method hair has been used as a bioindicator and investigated in this study. The relationship between intake and excretion in hair has been estimated in five volunteers who ingested radioactive polonium ((209)Po as a bio-tracer for (210)Po) in well determined quantities. Four of the volunteers were given 5-10 Bq (209)Po in a single intake (acute intake) and one volunteer has ingested a daily intake of 58.7 mBq (209)Po for a period of 180 d. Human hair was found to reflect the daily clearance of ingested polonium peaking at 0.001-0.01% d(-1) of the ingested amount, thereafter decreasing mono-exponentially, corresponding to a biological half-time of 10-20 days. For the case of protracted intake a mono-exponential build-up was observed with a half-time of 40 ± 5 d. In addition, after cessation of intake, a short-term component (74%) with a biological half-time of 16 ± 4 d, and a long-term component (26%) with a half-time of 93 ± 53 d were observed. It is concluded that hair can be used to detect not only the amount of ingested polonium but also whether the intake was protracted or acute.

  9. Purification, crystallization and preliminary X-ray diffraction analysis of the kinase domain of human tousled-like kinase 2

    SciTech Connect

    Garrote, Ana M.; Redondo, Pilar; Montoya, Guillermo; Muñoz, Inés G.

    2014-02-19

    The C-terminal kinase domain of TLK2 (a human tousled-like kinase) has been cloned and overexpressed in Escherichia coli followed by purification to homogeneity. Crystallization experiments in the presence of ATP-γ-S yielded crystals suitable for X-ray diffraction analysis belonging to two different space groups: tetragonal I4{sub 1}22 and cubic P2{sub 1}3. Tousled-like kinases (TLKs) are an evolutionarily conserved family of serine/threonine protein kinases involved in chromatin dynamics, including DNA replication and repair, transcription and chromosome segregation. The two members of the family reported in humans, namely TLK1 and TLK2, localize to the cell nucleus and are capable of forming homo- or hetero-oligomers by themselves. To characterize the role of TLK2, its C-terminal kinase domain was cloned and overexpressed in Escherichia coli followed by purification to homogeneity. Crystallization experiments in the presence of ATP-γ-S yielded crystals suitable for X-ray diffraction analysis belonging to two different space groups: tetragonal I4{sub 1}22 and cubic P2{sub 1}3. The latter produced the best diffracting crystal (3.4 Å resolution using synchrotron radiation), with unit-cell parameters a = b = c = 126.05 Å, α = β = γ = 90°. The asymmetric unit contained one protein molecule, with a Matthews coefficient of 4.59 Å{sup 3} Da{sup −1} and a solvent content of 73.23%.

  10. Crystallization and preliminary X-ray crystallographic analysis of human RGS10 complexed with Gαi3

    SciTech Connect

    Lee, Hyung Ki; Rhee, Kyung Hee; Kim, Chan-Wha; Hwang, Kwang Yeon; Kim, Eunice EunKyeong

    2005-09-01

    Human RGS10 and Gαi3 have been overexpressed and purified. Their complex has been crystallized (space group P4{sub 3}2{sub 1}2 or P4{sub 1}2{sub 1}2, unit-cell parameters a = 99.88, b = 99.88, c = 144.59 Å, α = β = γ = 90°) and diffraction data have been collected to 2.5 Å resolution using synchrotron radiation. G-protein-coupled receptors, which are major targets for drug discovery, play a major role in diverse physiological processes by relating changes in the extracellular environment to intracellular functions via activation of heterotrimeric G-proteins. However, G-protein activity is also modulated by a family of proteins called regulators of G-protein signalling (RGS), which are classified into six subfamilies. RGS10 belongs to the subgroup D/R12 and is known to act specifically on activated forms of three Gα proteins (Gαi3, Gαz and Gαo but not Gαs). It is abundantly expressed in brain and immune tissues and has been implicated in the pathophysiology of schizophrenia. The RGS domain of RGS10 was cloned, purified, complexed with human Gαi3 and crystallized. The crystals containing both RGS and Gαi3 belong to space group P4{sub 3}2{sub 1}2 (or P4{sub 1}2{sub 1}2), with unit-cell parameters a = 99.88, b = 99.88, c = 144.59 Å, α = β = γ = 90°. A full set of diffraction data were collected to 2.5 Å resolution at 100 K using synchrotron radiation at Pohang beamline 4A.

  11. Specific identification of human papillomavirus type in cervical smears and paraffin sections by in situ hybridization with radioactive probes: a preliminary communication

    SciTech Connect

    Gupta, J.; Gendelman, H.E.; Naghashfar, Z.; Gupta, P.; Rosenshein, N.; Sawada, E.; Woodruff, J.D.; Shah, K.

    1985-01-01

    Cervical Papanicolaou smears and paraffin sections of biopsy specimens obtained from women attending dysplasia clinics were examined for viral DNA sequences by in situ hybridization technique using TVS-labeled cloned recombinant DNA probes of human papillomavirus (HPV) types 6, 11, and 16. These and one unrelated DNA probe complementary to measles virus RNA were labeled by nick translation using either one or two TVS-labeled nucleotides. Paraffin sections and cervical smears were collected on pretreated slides, hybridized with the probes under stringent or nonstringent conditions for 50 h, and autoradiographed. Additional cervical specimens from the same women were examined for the presence of genus-specific papillomavirus capsid antigen by the immunoperoxidase technique. Preliminary results may be summarized as follows. The infecting virus could be identified in smears as well as in sections. Viral DNA sequences were detected only when there were condylomatous cells in the specimen and in only a proportion of the condylomatous cells. Even under stringent conditions, some specimens reacted with both HPV-6 and HPV-11. In some instances, the cells did not hybridize with any of the three probes even when duplicate specimens contained frankly condylomatous, capsid antigen-positive cells. In situ hybridization of Papanicolaou smears or of tissue sections is a practical method for diagnosis and follow-up of specific papillomavirus infection using routinely collected material.

  12. Preliminary study on non-viral transfection of F9 (factor IX) gene by nucleofection in human adipose-derived mesenchymal stem cells

    PubMed Central

    Olmedillas López, Susana; Garcia-Arranz, Mariano; Garcia-Olmo, Damian

    2016-01-01

    Background. Hemophilia is a rare recessive X-linked disease characterized by a deficiency of coagulation factor VIII or factor IX. Its current treatment is merely palliative. Advanced therapies are likely to become the treatment of choice for the disease as they could provide a curative treatment. Methods. The present study looks into the use of a safe non-viral transfection method based on nucleofection to express and secrete human clotting factor IX (hFIX) where human adipose tissue derived mesenchymal stem cells were used as target cells in vitro studies and NOD. Cg-Prkdcscid Il2rgtm1Wjl/SzJ mice were used to analyze factor IX expression in vivo studies. Previously, acute liver injury was induced by an injected intraperitoneal dose of 500 mg/kg body weight of acetaminophen. Results. Nucleofection showed a percentage of positive cells ranging between 30.7% and 41.9% and a cell viability rate of 29.8%, and cells were shown to secrete amounts of hFIX between 36.8 and 71.9 ng/mL. hFIX levels in the blood of NSG mice injected with ASCs transfected with this vector, were 2.7 ng/mL 48 h after injection. Expression and secretion of hFIX were achieved both in vitro cell culture media and in vivo in the plasma of mice treated with the transfected ASCs. Such cells are capable of eventually migrating to a previously damaged target tissue (the liver) where they secrete hFIX, releasing it to the bloodstream over a period of at least five days from administration. Conclusions. The results obtained in the present study may form a preliminary basis for the establishment of a future ex vivo non-viral gene/cellular safe therapy protocol that may eventually contribute to advancing the treatment of hemophilia. PMID:27114871

  13. Proteomic analyses of human cytomegalovirus strain AD169 derivatives reveal highly conserved patterns of viral and cellular proteins in infected fibroblasts.

    PubMed

    Reyda, Sabine; Büscher, Nicole; Tenzer, Stefan; Plachter, Bodo

    2014-01-07

    Human cytomegalovirus (HCMV) particle morphogenesis in infected cells is an orchestrated process that eventually results in the release of enveloped virions. Proteomic analysis has been employed to reveal the complexity in the protein composition of these extracellular particles. Only limited information is however available regarding the proteome of infected cells preceding the release of HCMV virions. We used quantitative mass spectrometry to address the pattern of viral and cellular proteins in cells, infected with derivatives of the AD169 laboratory strain. Our analyses revealed a remarkable conservation in the patterns of viral and of abundant cellular proteins in cells, infected for 2 hours, 2 days, or 4 days. Most viral proteins increased in abundance as the infection progressed over time. Of the proteins that were reliably detectable by mass spectrometry, only IE1 (pUL123), pTRS1, and pIRS1 were downregulated at 4 days after infection. In addition, little variation of viral proteins in the virions of the different viruses was detectable, independent of the expression of the major tegument protein pp65. Taken together these data suggest that there is little variation in the expression program of viral and cellular proteins in cells infected with related HCMVs, resulting in a conserved pattern of viral proteins ultimately associated with extracellular virions.

  14. Complete genome and gene expression analyses of Asaia bogorensis reveal unique responses to culture with mammalian cells as a potential opportunistic human pathogen.

    PubMed

    Kawai, Mikihiko; Higashiura, Norie; Hayasaki, Kimie; Okamoto, Naruhei; Takami, Akiko; Hirakawa, Hideki; Matsushita, Kazunobu; Azuma, Yoshinao

    2015-10-01

    Asaia bogorensis, a member of acetic acid bacteria (AAB), is an aerobic bacterium isolated from flowers and fruits, as well as an opportunistic pathogen that causes human peritonitis and bacteraemia. Here, we determined the complete genomic sequence of the As. bogorensis type strain NBRC 16594, and conducted comparative analyses of gene expression under different conditions of co-culture with mammalian cells and standard AAB culture. The genome of As. bogorensis contained 2,758 protein-coding genes within a circular chromosome of 3,198,265 bp. There were two complete operons encoding cytochrome bo3-type ubiquinol terminal oxidases: cyoABCD-1 and cyoABCD-2. The cyoABCD-1 operon was phylogenetically common to AAB genomes, whereas the cyoABCD-2 operon belonged to a lineage distinctive from the cyoABCD-1 operon. Interestingly, cyoABCD-1 was less expressed under co-culture conditions than under the AAB culture conditions, whereas the converse was true for cyoABCD-2. Asaia bogorensis shared pathogenesis-related genes with another pathogenic AAB, Granulibacter bethesdensis, including a gene coding pathogen-specific large bacterial adhesin and additional genes for the inhibition of oxidation and antibiotic resistance. Expression alteration of the respiratory chain and unique hypothetical genes may be key traits that enable the bacterium to survive under the co-culture conditions.

  15. A model for the development of human IgD-only B cells: Genotypic analyses suggest their generation in superantigen driven immune responses.

    PubMed

    Seifert, Marc; Steimle-Grauer, Susanne A; Goossens, Tina; Hansmann, Martin-Leo; Bräuninger, Andreas; Küppers, Ralf

    2009-02-01

    Human peripheral blood (PB) B cells expressing only IgD and tonsillar IgD-secreting plasma cells carry highly mutated V(H) genes and show preferential Iglambda usage. To further characterize these peculiar cells and gain insight into their generation, we analysed rearranged V(H) and V(L) genes of single IgD-only lambda(+) PB B cells and IgD(+) plasma cells from four individuals each. We demonstrate that the high somatic hypermutation activity in these cells is not restricted to V(H) genes but also present in V(L) genes. Moreover, not only PB IgD-only B cells, as reported earlier, but also IgD-expressing plasma cells often belong to very large clones. Surprisingly, the V(H)3-30 gene segment was used in each PB donor by >30% of IgD-only cells and in 2 tonsils by >50% of IgD plasma cells, whereas it was used less frequent in other B cells. All these features fit to a model in which IgD-only cells develop in superantigen-driven germinal center reactions, in which B cells are activated by binding of antigens to constant parts of Cdelta and often lambda light chains and the V(H)3-30 segment, and are selected for deletion of Cmu. IgD-only B cells may hence represent a unique B lineage subset generated in response to particular antigens.

  16. Complete genome and gene expression analyses of Asaia bogorensis reveal unique responses to culture with mammalian cells as a potential opportunistic human pathogen

    PubMed Central

    Kawai, Mikihiko; Higashiura, Norie; Hayasaki, Kimie; Okamoto, Naruhei; Takami, Akiko; Hirakawa, Hideki; Matsushita, Kazunobu; Azuma, Yoshinao

    2015-01-01

    Asaia bogorensis, a member of acetic acid bacteria (AAB), is an aerobic bacterium isolated from flowers and fruits, as well as an opportunistic pathogen that causes human peritonitis and bacteraemia. Here, we determined the complete genomic sequence of the As. bogorensis type strain NBRC 16594, and conducted comparative analyses of gene expression under different conditions of co-culture with mammalian cells and standard AAB culture. The genome of As. bogorensis contained 2,758 protein-coding genes within a circular chromosome of 3,198,265 bp. There were two complete operons encoding cytochrome bo3-type ubiquinol terminal oxidases: cyoABCD-1 and cyoABCD-2. The cyoABCD-1 operon was phylogenetically common to AAB genomes, whereas the cyoABCD-2 operon belonged to a lineage distinctive from the cyoABCD-1 operon. Interestingly, cyoABCD-1 was less expressed under co-culture conditions than under the AAB culture conditions, whereas the converse was true for cyoABCD-2. Asaia bogorensis shared pathogenesis-related genes with another pathogenic AAB, Granulibacter bethesdensis, including a gene coding pathogen-specific large bacterial adhesin and additional genes for the inhibition of oxidation and antibiotic resistance. Expression alteration of the respiratory chain and unique hypothetical genes may be key traits that enable the bacterium to survive under the co-culture conditions. PMID:26358298

  17. Phylogenetic and molecular analyses of human parainfluenza type 3 virus in Buenos Aires, Argentina, between 2009 and 2013: The emergence of new genetic lineages.

    PubMed

    Goya, Stephanie; Mistchenko, Alicia Susana; Viegas, Mariana

    2016-04-01

    Despite that human parainfluenza type 3 viruses (HPIV3) are one of the leading causes of acute lower respiratory tract infections in children under five, there is no licensed vaccine and there is limited current information on the molecular characteristics of regional and global circulating strains. The aim of this study was to describe the molecular characterization of HPIV3 circulating in Buenos Aires. We performed a genetic and phylogenetic analysis of the HN glycoprotein gene. Between 2009 and 2013, 124 HPIV3-positive samples taken from hospitalized pediatric patients were analyzed. Four new genetic lineages were described. Among them, C1c and C3d lineages showed local circulation patterns, whereas C3e and C3f comprised sequences from very distant countries. Despite the diversity of the described genotypes, C3a and C3d predominated over the others, the latter was present during the first years of the study and it was progressively replaced by C3a. Molecular analyses showed 28 non-synonymous substitutions; of these, 13 were located in potentially predicted B-cell epitopes. Taken together, the emergence of genetic lineages and the information of the molecular characteristics of HN protein may contribute to the general knowledge of HPIV3 molecular epidemiology for future vaccine development and antiviral therapies.

  18. Preliminary observations on polar body extrusion and pronuclear formation in human oocytes using time-lapse video cinematography.

    PubMed

    Payne, D; Flaherty, S P; Barry, M F; Matthews, C D

    1997-03-01

    In this study, we have used time-lapse video cinematography to study fertilization in 50 human oocytes that had undergone intracytoplasmic sperm injection (ICSI). Time-lapse recording commenced shortly after ICSI and proceeded for 17-20 h. Oocytes were cultured in an environmental chamber which was maintained under standard culture conditions. Overall, 38 oocytes (76%) were fertilized normally, and the fertilization rate and embryo quality were not significantly different from 487 sibling oocytes cultured in a conventional incubator. Normal fertilization followed a defined course of events, although the timing of these events varied markedly between oocytes. In 35 of the 38 fertilized oocytes (92%), there were circular waves of granulation within the ooplasm which had a periodicity of 20-53 min. The sperm head decondensed during this granulation phase. The second polar body was then extruded, and this was followed by the central formation of the male pronucleus. The female pronucleus formed in the cytoplasm adjacent to the second polar body at the same time as, or slightly after, the male pronucleus, and was subsequently drawn towards the male pronucleus until the two abutted. Both pronuclei then increased in size, the nucleoli moved around within the pronuclei and some nucleoli coalesced. During pronuclear growth, the organelles contracted from the cortex towards the centre of the oocyte, leaving a clear cortical zone. The oocyte decreased in diameter from 112 to 106 microm (P < 0.0001) during the course of the observation period. The female pronucleus was significantly smaller in diameter than the male pronucleus (24.1 and 22.4 microm respectively, P = 0.008) and contained fewer nucleoli (4.2 and 7.0 respectively, P < 0.0001). After time-lapse recording, oocytes were cultured for 48 h prior to embryo transfer or cryopreservation. Embryo quality was related to fertilization events and periodicity of the cytoplasmic wave, and it was found that good quality embryos

  19. Exposure to Hexabromocyclododecanes (HBCDs) via Dust Ingestion, but Not Diet, Correlates with Concentrations in Human Serum: Preliminary Results

    PubMed Central

    Roosens, Laurence; Abdallah, Mohamed Abou-Elwafa; Harrad, Stuart; Neels, Hugo; Covaci, Adrian

    2009-01-01

    Background Hexabromocyclododecane (HBCD) is a high-production-volume chemical used as flame retardant in polystyrene insulation and textiles. Because it is not chemically bound to the polymer, HBCD can migrate into the environment, contaminating indoor dust and foodstuff. Objectives We examined for the first time the relationship between combined exposure to three HBCD isomers (∑HBCDs) via ingestion of food (duplicate diets) and indoor dust and HBCD concentrations in serum for 16 Belgian adults (20–25 years of age). We also determined the chiral signatures of HBCDs to advance understanding of source-to-human enantioselective degradation and/or metabolism. Methods Concentrations and chiral signatures of α-, β-, and γ-HBCD in duplicate diets, dust, and serum were measured by liquid chromatography/tandem mass spectrometry. Results Dietary intakes of ∑HBCDs were 1.2–20 ng/day (average, 7.2 ng/day), whereas those estimated under average (20 mg dust/day) and high (50 mg dust/day) dust ingestion scenarios were 1.1–15 ng/day (average intake, 3.2 ng/day) and 2.8–38 ng/day (average intake, 8.0 ng/day), respectively. Concentrations of ∑HBCDs measured in blood serum were < 0.5 to 11 ng/g lipid weight (lw) (average, 2.9 ng/g lw). γ-HBCD dominated in food, whereas α-HBCD dominated in dust and was the sole isomer in serum. Although exposure via dust ingestion correlated significantly (p < 0.01) with concentrations in serum, no such correlation was evident with dietary exposure (p > 0.1). Although no enantioselective enrichment was detected in either dust or diet, substantial enrichment of (−)α-HBCD was observed in serum. Conclusions Serum concentrations of HBCDs were correlated with the exposure via dust, but not via dietary ingestion. The enrichment of the (−)α-HBCD enantiomer in humans appears to be due to in vivo enantioselective metabolism/excretion rather than ingestion of dust or diet. PMID:20049121

  20. Crystallization and preliminary X-ray characterization of 1,3-propanediol dehydrogenase from the human pathogen Klebsiella pneumoniae

    SciTech Connect

    Marçal, D.; Rego, A. T.; Fogg, M. J.; Wilson, K. S.; Carrondo, M. A.; Enguita, F. J.

    2007-03-01

    1,3-Propanediol dehydrogenase from K. pneumoniae has been overexpressed in E. coli, purified and crystallized. Diffraction data have been collected to 2.7 Å resolution. 1,3-Propanediol dehydrogenase (1,3-PD-DH), encoded by the dhaT gene, is a key enzyme in the dissimilation process for converting glycerol to 1,3-propanediol in the human pathogen Klebsiella pneumoniae. Single colourless crystals were obtained from a recombinant preparation of 1,3-propanediol dehydrogenase overexpressed in Escherichia coli. The crystals belong to space group P2{sub 1}, with unit-cell parameters a = 91.9, b = 226.6, c = 232.6 Å, β = 92.9°. The crystals probably contain two decamers in the asymmetric unit, with a V{sub M} value of 3.07 Å{sup 3} Da{sup −1} and an estimated solvent content of 59%. Diffraction data were collected to 2.7 Å resolution using synchrotron radiation at the ID14-4 beamline of the European Synchrotron Radiation Facility.

  1. Purification, crystallization and preliminary X-ray diffraction analysis of the kinase domain of human tousled-like kinase 2

    PubMed Central

    Garrote, Ana M.; Redondo, Pilar; Montoya, Guillermo; Muñoz, Inés G.

    2014-01-01

    Tousled-like kinases (TLKs) are an evolutionarily conserved family of serine/threonine protein kinases involved in chromatin dynamics, including DNA replication and repair, transcription and chromosome segregation. The two members of the family reported in humans, namely TLK1 and TLK2, localize to the cell nucleus and are capable of forming homo- or hetero-oligomers by themselves. To characterize the role of TLK2, its C-terminal kinase domain was cloned and overexpressed in Escherichia coli followed by purification to homogeneity. Crystallization experiments in the presence of ATP-γ-S yielded crystals suitable for X-ray diffraction analysis belonging to two different space groups: tetragonal I4122 and cubic P213. The latter produced the best diffracting crystal (3.4 Å resolution using synchrotron radiation), with unit-cell parameters a = b = c = 126.05 Å, α = β = γ = 90°. The asymmetric unit contained one protein molecule, with a Matthews coefficient of 4.59 Å3 Da−1 and a solvent content of 73.23%. PMID:24598926

  2. Preliminary human health risk assessment of arsenic and fluoride in tap water from Zacatecas, México.

    PubMed

    Martínez-Acuña, Mónica I; Mercado-Reyes, Marisa; Alegría-Torres, Jorge A; Mejía-Saavedra, José J

    2016-08-01

    Zacatecas state is located in the central area of Mexico, where the underground water contains elevated quantities of natural arsenic and fluoride. In order to estimate health risk associated with human exposure to these pollutants, tap water samples from the southern-central region of the state were analyzed. Ninety percent of the samples exceeded the levels of arsenic established by the World Health Organization (WHO) of 0.01 mg/L and 43 % exceeded the limit established by the NOM-127-SSA1(1) of 0.025 mg/L. Forty-three percent of the samples had fluoride levels above the Mexican regulation limit of 1.5 mg/L (NOM-127-SSA1). We used WHO and EPA's health risk assessment method, we estimated 80 % of the inhabitants of sites studied could be exposed to arsenic levels higher than those recommended by EPA and the WHO, 22 % could be exposed to fluoride levels higher than those recommended by EPA, and 16 % of the local population may be in risk of suffering dental fluorosis.

  3. Crystallization and preliminary crystallographic analysis of orotidine 5′-monophosphate decarboxylase from the human malaria parasite Plasmodium falciparum

    SciTech Connect

    Krungkrai, Sudaratana R.; Tokuoka, Keiji; Kusakari, Yukiko; Inoue, Tsuyoshi; Adachi, Hiroaki; Matsumura, Hiroyoshi; Takano, Kazufumi; Murakami, Satoshi; Mori, Yusuke; Kai, Yasushi; Krungkrai, Jerapan; Horii, Toshihiro

    2006-06-01

    Orotidine 5′-monophosphate decarboxylase of human malaria parasite P. falciparum was crystallized by the seeding method in a hanging drop using PEG 3000 as a precipitant. A complete set of diffraction data from a native crystal was collected to 2.7 Å resolution at 100 K using synchrotron radiation. Orotidine 5′-monophosphate (OMP) decarboxylase (OMPDC; EC 4.1.1.23) catalyzes the final step in the de novo synthesis of uridine 5′-monophosphate (UMP) and defects in the enzyme are lethal in the malaria parasite Plasmodium falciparum. Active recombinant P. falciparum OMPDC (PfOMPDC) was crystallized by the seeding method in a hanging drop using PEG 3000 as a precipitant. A complete set of diffraction data from a native crystal was collected to 2.7 Å resolution at 100 K using synchrotron radiation at the Swiss Light Source. The crystal exhibits trigonal symmetry (space group R3), with hexagonal unit-cell parameters a = b = 201.81, c = 44.03 Å. With a dimer in the asymmetric unit, the solvent content is 46% (V{sub M} = 2.3 Å{sup 3} Da{sup −1})

  4. Feasibility study and preliminary design of load-assisting clothes for lumbar protection inspired by human musculoskeletal systems

    NASA Astrophysics Data System (ADS)

    Hashimoto, Riho; Masuda, Arata; Chen, Hao; Kobayashi, Sou

    2016-04-01

    The purpose of this paper is to develop load assisting clothes for caregivers. Low back pain is one of the most major reasons for caregivers to leave their jobs. In this study, load assisting clothes which reduce the risks of low back pain of caregivers are designed and manufactured, targeting at the use in small care-houses and family caregiving. The load assisting clothes should have two functions. One is to reduce the compressive load acting on the lumbar spine as well as the tensile load on the lumbar muscles by providing an appropriate assisting force. The other is not to interfere with wearers' motion. The proposed approach in this study is to put elastic compressive members and tensioner belts integrated in the garment to provide the assisting forces without hindering natural movement and comfortable feeling. We study human musculoskeletal systems in the lumbar part, and consider to construct a parallel reinforcement of it on the body surface by embedding passive support structures. The arrangement of those elements is determined based on the study of the principal strain directions and the non-extension directions of the body surface to manage the appropriate assisting force without spoiling the mobility. The effectiveness of the proposed support principle is verified through experimental studies.

  5. A preliminary analysis of within-subject variation in human serum oxidative stress parameters as a function of time.

    PubMed

    Argüelles, Sandro; Gómez, Ana; Machado, Alberto; Ayala, Antonio

    2007-12-01

    The aim of this study was to determine variability at both levels of two serum oxidative stress markers (lipid peroxides and carbonyl concentration) as well as total antioxidant capacity in humans as a function of time. Assays for oxidative stress and antioxidant capacity were repeated in the same individuals three times daily on four particular days over a period of 51 days. The results show a high variation within subject in the concentration of these markers not only when comparing the different days (the morning values can change up to 98%), but also during the day, where the evening values can increase up to 84% with respect to those of the morning. This suggests that several measurements are required to establish the typical oxidative stress status of an individual before studying the potential effect of treatments that possibly influence oxidative damage. The observed changes during the day allowed us to speculate about the optimum temporal antioxidant delivery regimes that minimize the imbalance between oxidants and antioxidants. In the study, only a few general aspects of basic lifestyle habits were controlled. However, the levels of these markers are sensitive to possibly a group of factors. This points to the necessity of using a much bigger population to establish the possible contribution of each lifestyle habits to the concentration of the markers.

  6. Crystallization and preliminary X-ray diffraction analysis of the ternary human GM-CSF receptor complex

    SciTech Connect

    Hansen, Guido; Hercus, Timothy R.; Xu, Yibin; Lopez, Angel F.; Parker, Michael W.; McKinstry, William J.

    2008-07-28

    Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a haemopoietic growth factor that acts through a ternary receptor signalling complex containing specific {alpha} (GMR{alpha}) and common {beta} ({beta}c) receptor subunits. Human GM-CSF is encoded by the gene csf2, while the genes for GMR{alpha} and {beta}c are csf2ra and csf2rb, respectively. Crystals of the ternary ectodomain complex comprising GM-CSF and the soluble extracellular regions of both the GMR{alpha} subunit and either {beta}c or its glutamine-substitution mutant N346Q were obtained using the hanging-drop vapour-diffusion method. The best diffracting crystals of the ternary complex were obtained using the N346Q mutation of the {beta}c subunit. These crystals grew using polyethylene glycol 3350 with a high concentration of proline, belonged to space group P6{sub 3}22 and diffracted to 3.3 {angstrom} resolution.

  7. Bilateral sensory deprivation of trigeminal afferent fibres on corticomotor control of human tongue musculature: a preliminary study.

    PubMed

    Kothari, M; Baad-Hansen, L; Svensson, P

    2016-09-01

    Transcranial magnetic stimulation (TMS) has demonstrated changes in motor evoked potentials (MEPs) in human limb muscles following modulation of sensory afferent inputs. The aim of this study was to determine whether bilateral local anaesthesia (LA) of the lingual nerve affects the excitability of the tongue motor cortex (MI) as measured by TMS. The effect on MEPs after bilateral LA of the lingual nerve was studied, while the first dorsal interosseous (FDI) muscle served as a control in ten healthy participants. MEPs were measured on the right side of the tongue dorsum in four different conditions: (i) immediately prior to anaesthesia (baseline), (ii) during bilateral LA block of the lingual nerve, (iii) after anaesthesia had subjectively subsided (recovery) and (iv) 3 h after bilateral lingual block injection. MEPs were assessed using stimulus-response curves in steps of 10% of motor threshold (T). Eight stimuli were given at each stimulus level. The amplitudes of the tongue MEPs were significantly influenced by the stimulus intensity (P < 0·001) but not by condition (P = 0·186). However, post hoc tests showed that MEPS were statistically significantly higher during bilateral LA block condition compared with baseline at T + 40%, T + 50% and T + 60% (P < 0·028) and also compared with recovery at T + 60% (P = 0·010) as well as at 3 h after injection at T + 50% and T + 60% (P < 0·029). Bilateral LA block of the lingual nerve seems to be associated with a facilitation of the corticomotor pathways related to the tongue musculature.

  8. The effects and risks associated to mephedrone and methylone in humans: A review of the preliminary evidences.

    PubMed

    Karila, Laurent; Billieux, Joel; Benyamina, Amine; Lançon, Christophe; Cottencin, Olivier

    2016-09-01

    New psychoactive substances have drastically modified the world drug scene. An increasingly popular class comprises synthetic or substituted cathinones (legal highs, research chemicals, bath salts). Among the most common psychoactive constituents of bath salts are mephedrone and methylone. Recent reports on the abuse of novel synthetic cathinone derivatives call attention to the serious physical and psychological risks resulting from their consumption, thereby emphasizing the growing use of these drugs might constitute an important public health issue. In this paper, we will review the available data regarding the use and effects of mephedrone and methylone in humans in order to highlight their impact on public health. To reach this objective, a literature search was performed on two representative databases (Pubmed, Google Scholar), the Erowid Center website (a US non-profit educational organization that provides information about psychoactive plants and chemicals), and various governmental websites. The terms used for the database search were "mephedrone", "methylone", "new psychoactive substances", "synthetic cathinones", "substituted cathinones", "substance abuse", "substance use disorder", "adverse effects", "fatalities". The literature search was limited to years 2005-2015 and led to the identification of 71 potentially relevant articles. To date, the actual prevalence rates of their use remains difficult to estimate. Important health-related issues have emerged in relation to the somatic, psychiatric, and addictive consequences of their use. The potential chronic health effects of their prolonged use remain to date unknown (e.g., reproductive toxicity, genotoxicity and carcinogenic potential). Treatment for patients with prolonged exposure to synthetic cathinones should ideally include a drug management plan coupled with psychotherapy taking place in a structured program of care.

  9. Articular cartilage repair with recombinant human type II collagen/polylactide scaffold in a preliminary porcine study.

    PubMed

    Muhonen, Virpi; Salonius, Eve; Haaparanta, Anne-Marie; Järvinen, Elina; Paatela, Teemu; Meller, Anna; Hannula, Markus; Björkman, Mimmi; Pyhältö, Tuomo; Ellä, Ville; Vasara, Anna; Töyräs, Juha; Kellomäki, Minna; Kiviranta, Ilkka

    2016-05-01

    The purpose of this study was to investigate the potential of a novel recombinant human type II collagen/polylactide scaffold (rhCo-PLA) in the repair of full-thickness cartilage lesions with autologous chondrocyte implantation technique (ACI). The forming repair tissue was compared to spontaneous healing (spontaneous) and repair with a commercial porcine type I/III collagen membrane (pCo). Domestic pigs (4-month-old, n = 20) were randomized into three study groups and a circular full-thickness chondral lesion with a diameter of 8 mm was created in the right medial femoral condyle. After 3 weeks, the chondral lesions were repaired with either rhCo-PLA or pCo together with autologous chondrocytes, or the lesion was only debrided and left untreated for spontaneous repair. The repair tissue was evaluated 4 months after the second operation. Hyaline cartilage formed most frequently in the rhCo-PLA treatment group. Biomechanically, there was a trend that both treatment groups resulted in better repair tissue than spontaneous healing. Adverse subchondral bone reactions developed less frequently in the spontaneous group (40%) and the rhCo-PLA treated group (50%) than in the pCo control group (100%). However, no statistically significant differences were found between the groups. The novel rhCo-PLA biomaterial showed promising results in this proof-of-concept study, but further studies will be needed in order to determine its effectiveness in articular cartilage repair. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:745-753, 2016.

  10. Impact of public programs on fertility and gender specific investment in human capital of children in rural India: cross sectional and time series analyses.

    PubMed

    Duraisamy, P; Malathy, R

    1991-01-01

    Cross sectional and time series analyses are conducted with 1971 and 1981 rural district level data for India in order to estimate variations in program impacts on household decisionmaking concerning fertility, child mortality, and schooling; to analyze how the variation in public program subsidies and services influences sex specific investments in schooling; and to examine the bias in cross sectional estimates by employing fixed effects methodology. The theory of household production uses the framework development by Rosenzweig and Wolpin. The utility function is expressed as a function of families' desired number of children, sex specific investment in human capital of children measured by schooling of males and females, and a composite consumption good. Budget constraints are characterized in terms of the biological supply of births or natural fertility, the number of births averted by fertility control, exogenous money income, the prices of number of children, contraceptives, child schooling, and consumption of goods. Demand functions are constructed from maximizing the utility function subject to the budget constraint. Data constitute 40% of the total districts and 50% of the rural population. The empirical specification of the linear model and variable description are provided. Other explanatory variables included are adult educational attainment; % of scheduled castes and tribes and % Muslim; and % rural population. Estimation methods are described and justification is provided for the use of ordinary least squares and fixed effects methods. The results of the cross sectional analysis reveal that own-program effects of family planning and primary health centers reduced family size in 1971 and 81. The increase in secondary school enrollment is evidenced in only 1971. There is a significant effect of family planning (FP) clinics on the demand for surviving children only in 1971. The presence of a seconary school in a village reduces the demand for children in

  11. Graphical, kinetic, and equilibrium analyses of in vivo [123I] beta-CIT binding to dopamine transporters in healthy human subjects.

    PubMed

    Laruelle, M; Wallace, E; Seibyl, J P; Baldwin, R M; Zea-Ponce, Y; Zoghbi, S S; Neumeyer, J L; Charney, D S; Hoffer, P B; Innis, R B

    1994-11-01

    The in vivo kinetics of the dopamine (DA) transporter probe 123I-labeled 2 beta-carboxymethoxy-3 beta-(4-iodophenyl) tropane ([123I] beta-CIT) in striatum was investigated with single-photon emission computerized tomography (SPECT) in five healthy human subjects. The aim of this study was to derive an adequate measure of the DA transporter density that would not be affected by regional cerebral blood flow or peripheral clearance of the tracer. SPECT data were acquired on the day of injection (day 1) from 0 to 7 h and on the following day (day 2) from 19 to 25 h. Arterial sampling on day 1 was used to measure the input function. Graphical, kinetic, and equilibrium analyses were evaluated. Graphical analysis of day 1 data, with the assumption of negligible dissociation of the tracer-receptor complex (k4 = 0), was found to be blood flow-dependent. A three-compartment kinetic analysis of day 1 data were performed using a three (k4 = 0)- and a four (k4 > 0)-parameter model. The three-parameter model estimated the konBmax product at 0.886 +/- 0.087 min-1. The four-parameter model gave a binding potential (BP) of 476 ml g-1, a value consistent with in vitro measurements. The stability of the regional uptake on day 2 allowed direct measurement of the specific to nonspecific equilibrium partition coefficient (V3" = k3/k4 = 6.66 +/- 1.54). Results of day 1 kinetic analysis and day 2 equilibrium analysis were well correlated among subjects. Simulations indicated that the error associated with the day 2 equilibrium analysis was acceptable for plasma tracer terminal half-lives > 10 h. We propose the equilibrium analysis on day 2 as the method of choice for clinical studies since it does not require multiple scans or the measurement of the arterial plasma tracer concentrations.

  12. Histology-driven data mining of lipid signatures from multiple imaging mass spectrometry analyses: application to human colorectal cancer liver metastasis biopsies.

    PubMed

    Thomas, Aurélien; Patterson, Nathan Heath; Marcinkiewicz, Martin M; Lazaris, Anthoula; Metrakos, Peter; Chaurand, Pierre

    2013-03-05

    Imaging mass spectrometry (IMS) represents an innovative tool in the cancer research pipeline, which is increasingly being used in clinical and pharmaceutical applications. The unique properties of the technique, especially the amount of data generated, make the handling of data from multiple IMS acquisitions challenging. This work presents a histology-driven IMS approach aiming to identify discriminant lipid signatures from the simultaneous mining of IMS data sets from multiple samples. The feasibility of the developed workflow is evaluated on a set of three human colorectal cancer liver metastasis (CRCLM) tissue sections. Lipid IMS on tissue sections was performed using MALDI-TOF/TOF MS in both negative and positive ionization modes after 1,5-diaminonaphthalene matrix deposition by sublimation. The combination of both positive and negative acquisition results was performed during data mining to simplify the process and interrogate a larger lipidome into a single analysis. To reduce the complexity of the IMS data sets, a sub data set was generated by randomly selecting a fixed number of spectra from a histologically defined region of interest, resulting in a 10-fold data reduction. Principal component analysis confirmed that the molecular selectivity of the regions of interest is maintained after data reduction. Partial least-squares and heat map analyses demonstrated a selective signature of the CRCLM, revealing lipids that are significantly up- and down-regulated in the tumor region. This comprehensive approach is thus of interest for defining disease signatures directly from IMS data sets by the use of combinatory data mining, opening novel routes of investigation for addressing the demands of the clinical setting.

  13. Extensive temporal transcriptome and microRNA analyses identify molecular mechanisms underlying mitochondrial dysfunction induced by multi-walled carbon nanotubes in human lung cells.

    PubMed

    Nymark, Penny; Wijshoff, Peter; Cavill, Rachel; van Herwijnen, Marcel; Coonen, Maarten L J; Claessen, Sandra; Catalán, Julia; Norppa, Hannu; Kleinjans, Jos C S; Briedé, Jacob J

    2015-01-01

    Understanding toxicity pathways of engineered nanomaterials (ENM) has recently been brought forward as a key step in twenty-first century ENM risk assessment. Molecular mechanisms linked to phenotypic end points is a step towards the development of toxicity tests based on key events, which may allow for grouping of ENM according to their modes of action. This study identified molecular mechanisms underlying mitochondrial dysfunction in human bronchial epithelial BEAS 2B cells following exposure to one of the most studied multi-walled carbon nanotubes (Mitsui MWCNT-7). Asbestos was used as a positive control and a non-carcinogenic glass wool material was included as a negative fibre control. Decreased mitochondrial membrane potential (MMP↓) was observed for MWCNTs at a biologically relevant dose (0.25 μg/cm(2)) and for asbestos at 2 μg/cm(2), but not for glass wool. Extensive temporal transcriptomic and microRNA expression analyses identified a 330-gene signature (including 26 genes with known mitochondrial function) related to MWCNT- and asbestos-induced MMP↓. Forty-nine of the MMP↓-associated genes showed highly similar expression patterns over time (six time points) and the majority was found to be regulated by two transcription factors strongly involved in mitochondrial homeostasis, APP and NRF1. In addition, four miRNAs were correlated with MMP↓ and one of them, miR-1275, was found to negatively correlate with a large part of the MMP↓-associated genes. Cellular processes such as gluconeogenesis, mitochondrial LC-fatty acid β-oxidation and spindle microtubule function were enriched among the MMP↓-associated genes and miRNAs. These results are expected to be useful in the identification of key events in ENM-related toxicity pathways for the development of molecular screening techniques.

  14. Transcriptome-Guided Functional Analyses Reveal Novel Biological Properties and Regulatory Hierarchy of Human Embryonic Stem Cell-Derived Ventricular Cardiomyocytes Crucial for Maturation

    PubMed Central

    Poon, Ellen; Yan, Bin; Zhang, Shaohong; Rushing, Stephanie; Keung, Wendy; Ren, Lihuan; Lieu, Deborah K.; Geng, Lin; Kong, Chi-Wing; Wang, Jiaxian; Wong, Hau San; Boheler, Kenneth R.; Li, Ronald A.

    2013-01-01

    Abstract Human (h) embryonic stem cells (ESC) represent an unlimited source of cardiomyocytes (CMs); however, these differentiated cells are immature. Thus far, gene profiling studies have been performed with non-purified or non-chamber specific CMs. Here we took a combinatorial approach of using systems biology to guide functional discoveries of novel biological properties of purified hESC-derived ventricular (V) CMs. We profiled the transcriptomes of hESCs, hESC-, fetal (hF) and adult (hA) VCMs, and showed that hESC-VCMs displayed a unique transcriptomic signature. Not only did a detailed comparison between hESC-VCMs and hF-VCMs confirm known expression changes in metabolic and contractile genes, it further revealed novel differences in genes associated with reactive oxygen species (ROS) metabolism, migration and cell cycle, as well as potassium and calcium ion transport. Following these guides, we functionally confirmed that hESC-VCMs expressed IKATP with immature properties, and were accordingly vulnerable to hypoxia/reoxygenation-induced apoptosis. For mechanistic insights, our coexpression and promoter analyses uncovered a novel transcriptional hierarchy involving select transcription factors (GATA4, HAND1, NKX2.5, PPARGC1A and TCF8), and genes involved in contraction, calcium homeostasis and metabolism. These data highlight novel expression and functional differences between hESC-VCMs and their fetal counterparts, and offer insights into the underlying cell developmental state. These findings may lead to mechanism-based methods for in vitro driven maturation. PMID:24204964

  15. Genetic analyses reveal differences in the VP7 and VP4 antigenic epitopes between human rotaviruses circulating in Belgium and rotaviruses in Rotarix and RotaTeq.

    PubMed

    Zeller, Mark; Patton, John T; Heylen, Elisabeth; De Coster, Sarah; Ciarlet, Max; Van Ranst, Marc; Matthijnssens, Jelle

    2012-03-01

    Two live-attenuated rotavirus group A (RVA) vaccines, Rotarix (G1P[8]) and RotaTeq (G1-G4, P[8]), have been successfully introduced in many countries worldwide, including Belgium. The parental RVA strains used to generate the vaccines were isolated more than 20 years ago in France (G4 parental strain in RotaTeq) and the United States (all other parental strains). At present, little is known about the relationship between currently circulating human RVAs and the vaccine strains. In this study, we determined sequences for the VP7 and VP4 outer capsid proteins of representative G1P[8], G2P[4], G3P[8], G4P[8], G9P[8], and G12P[8] RVAs circulating in Belgium during 2007 to 2009. The analyses showed that multiple amino acid differences existed between the VP7 and VP4 antigenic epitopes of the vaccine viruses and the Belgian isolates, regardless of their G and P genotypes. However, the highest variability was observed among the circulating G1P[8] RVA strains and the G1 and P[8] components of both RVA vaccines. In particular, RVA strains of the P[8] lineage 4 (OP354-like) showed a significant number of amino acid differences with the P[8] VP4 of both vaccines. In addition, the circulating Belgian G3 RVA strains were found to possibly possess an extra N-linked glycosylation site compared to the G3 RVA vaccine strain of RotaTeq. These results indicate that the antigenic epitopes of RVA strains contained in the vaccines differ substantially from those of the currently circulating RVA strains in Belgium. Over time, these differences might result in selection for strains that escape the RVA neutralizing-antibody pressure induced by vaccines.

  16. Functional analyses of human and zebrafish 18-amino acid in-frame deletion pave the way for domain mapping of the cerebral cavernous malformation 3 protein.

    PubMed

    Voss, Katrin; Stahl, Sonja; Hogan, Benjamin M; Reinders, Joerg; Schleider, Elisa; Schulte-Merker, Stefan; Felbor, Ute

    2009-06-01

    Cerebral cavernous malformations (CCMs) may cause recurrent headaches, seizures, and hemorrhagic stroke and have been associated with loss-of-function mutations in CCM1/KRIT1, CCM2, and CCM3/programmed cell death 10 (PDCD10). The CCM3/PDCD10 amino acid sequence does not reveal significant homologies to protein domains with known structure. With the help of the only published human in-frame deletion of the CCM3 gene (c.97-?_150+?del), CCM3:p.L33_K50del, we have identified the interaction domain of CCM3 with the oxidant stress response serine/threonine kinase 25 (STK25, YSK1, SOK1) and with the mammalian Ste20-like kinase 4 (MST4, MASK). Consistently, nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS) analyses revealed two STK25 phosphorylation sites at serine 39 and threonine 43. The corresponding in-frame deletion of zebrafish ccm3a, dccm3:p.L31_K48del, also resulted in impaired interaction with STK25 and MST4. In agreement with the observed redundant biochemical functionality of zebrafish ccm3a and its duplicate ccm3b, simultaneous inactivation of both genes resulted in a progressive cardiovascular phenotype in zebrafish indistinguishable from ccm1 and ccm2 mutants. The pronounced cardiovascular dilatations could be recapitulated by morpholino-induced in-frame skipping of the exon encoding the STK25 and MST4 binding site of zebrafish Ccm3a if Ccm3b was repressed in parallel. Using a novel zebrafish model of CCM, we could thus demonstrate that the newly mapped STK25 and MST4 interaction domain within the CCM3 protein plays a crucial role for vascular development in zebrafish.

  17. Identification and quantification of nicotine biomarkers in human oral fluid from individuals receiving low-dose transdermal nicotine: a preliminary study.

    PubMed

    Miller, Eleanor I; Norris, Hye-Ryun K; Rollins, Douglas E; Tiffany, Stephen T; Moore, Christine M; Vincent, Michael J; Agrawal, Alpana; Wilkins, Diana G

    2010-09-01

    The objective of this preliminary study was to identify and quantify potential nicotine (NIC) biomarkers in post-exposure oral fluid samples collected from 10 NIC-abstinent human participants administered 7 mg transdermal NIC using liquid chromatography-tandem mass spectrometry (LC-MS-MS). Oral fluid samples were collected prior to NIC patch application and at 0.5 and 0.75 h after patch removal using the Quantisal() oral fluid collection device. The validated LC-MS-MS analyte panel included nicotine-Nbeta-D-glucuronide, cotinine-N-oxide, trans-3-hydroxycotinine, norcotinine, trans-nicotine-1'-N-oxide, cotinine (COT), nornicotine, NIC, anatabine, anabasine, and cotinine-N-beta-D-glucuronide. Analytes and corresponding deuterated internal standards were extracted by solid-phase extraction. NIC and COT concentrations were quantifiable in oral fluid samples collected from 6 of the 10 participants 0.5 h after patch removal and in oral fluid samples collected from 7 of the 10 participants 0.75 h after patch removal. Based on the mean NIC and COT concentrations in oral fluid and plasma for the participants with both quantifiable NIC and COT at the 0.5 and 0.75 h collection times, the oral fluid-plasma ratio was 6.4 for NIC and 3.3 for COT. An ELISA procedure was also validated and successfully applied as a screening tool for these oral fluid samples in conjunction with LC-MS-MS confirmation. An ELISA cut-off concentration of 5.0 ng/mL provided excellent sensitivity for discrimination of COT-positive post-exposure oral fluid samples collected after low-level transdermal NIC exposure and oral fluid samples collected prior to patch application.

  18. Complete Genome Sequence of Germline Chromosomally Integrated Human Herpesvirus 6A and Analyses Integration Sites Define a New Human Endogenous Virus with Potential to Reactivate as an Emerging Infection.

    PubMed

    Tweedy, Joshua; Spyrou, Maria Alexandra; Pearson, Max; Lassner, Dirk; Kuhl, Uwe; Gompels, Ursula A

    2016-01-15

    Human herpesvirus-6A and B (HHV-6A, HHV-6B) have recently defined endogenous genomes, resulting from integration into the germline: chromosomally-integrated "CiHHV-6A/B". These affect approximately 1.0% of human populations, giving potential for virus gene expression in every cell. We previously showed that CiHHV-6A was more divergent than CiHHV-6B by examining four genes in 44 European CiHHV-6A/B cardiac/haematology patients. There was evidence for gene expression/reactivation, implying functional non-defective genomes. To further define the relationship between HHV-6A and CiHHV-6A we used next-generation sequencing to characterize genomes from three CiHHV-6A cardiac patients. Comparisons to known exogenous HHV-6A showed CiHHV-6A genomes formed a separate clade; including all 85 non-interrupted genes and necessary cis-acting signals for reactivation as infectious virus. Greater single nucleotide polymorphism (SNP) density was defined in 16 genes and the direct repeats (DR) terminal regions. Using these SNPs, deep sequencing analyses demonstrated superinfection with exogenous HHV-6A in two of the CiHHV-6A patients with recurrent cardiac disease. Characterisation of the integration sites in twelve patients identified the human chromosome 17p subtelomere as a prevalent site, which had specific repeat structures and phylogenetically related CiHHV-6A coding sequences indicating common ancestral origins. Overall CiHHV-6A genomes were similar, but distinct from known exogenous HHV-6A virus, and have the capacity to reactivate as emerging virus infections.

  19. High-speed flow cytometric analysis of nanoparticle targeting to rare leukemic stem cells in peripheral human blood: preliminary in-vitro studies

    NASA Astrophysics Data System (ADS)

    Cooper, Christy L.; Leary, James F.

    2014-03-01

    Leukemic cancer stem cells are both stem-like and leukemic-like. This complicates their detection as rare circulating tumor cells in peripheral blood of leukemia patients. The leukemic stem cells are also highly resistant to standard chemotherapeutic regimens so new therapeutic strategies need to be designed to kill the leukemic stem cells without killing normal stem cells. In these initial studies we have designed an antibody-targeted and fluorescent (Cy5.5) nanoparticle for targeting these leukemic stem cells and then introducing new strategies for killing them. Multicolor flow cytometric analyses were performed on a BD FACS Aria III. Human leukemic stem cell-like cell line RS4;11 (with putative immunophenotype CD123+/CD24+/CD38-/CD10-/Flt-3-) was used as a model human leukemic stem cell systems and were spiked into normal human peripheral blood cells containing normal blood stem-progenitor cells (immunophenotype CD123-/CD34+/CD38-) and Cy5.5-labeled nanoparticles with targeting molecule anti-CD123 antibody. An irrelevant antibody (CD71) which should not bind to any live leukemic stem cell or normal stem cell (binds erythrocytes) was used as a way of distinguishing between true-positive live and false-positive damaged/dead cells, the latter occurring at much higher frequencies than the very rare (e.g. 0.001 to 0.0001 percent frequency true leukemic stem cells). These studies are designed to measure the targeting sensitivity and specificity of the fluorescent nanoparticles to the putative rare leukemic stem cells with the eventual design to use the nanoparticles to direct killing therapeutic doses to the leukemic stem cells but not to the normal stem-progenitor cells.

  20. San Mateo Creek Basin Preliminary Assessment

    EPA Pesticide Factsheets

    The objective of this Preliminary Assessment is to evaluate the site using the Hazard Ranking System and the Superfund Chemical Data Matrix to determine if a threat to human health and the environment exists such that further action is warranted.

  1. Relaxation-compensated CEST-MRI at 7 T for mapping of creatine content and pH--preliminary application in human muscle tissue in vivo.

    PubMed

    Rerich, Eugenia; Zaiss, Moritz; Korzowski, Andreas; Ladd, Mark E; Bachert, Peter

    2015-11-01

    The small biomolecule creatine is involved in energy metabolism. Mapping of the total creatine (mostly PCr and Cr) in vivo has been done with chemical shift imaging. Chemical exchange saturation transfer (CEST) allows an alternative detection of creatine via water MRI. Living tissue exhibits CEST effects from different small metabolites, including creatine, with four exchanging protons of its guanidinium group resonating about 2 ppm from the water peak and hence contributing to the amine proton CEST peak. The intermediate exchange rate (≈ 1000 Hz) of the guanidinium protons requires high RF saturation amplitude B1. However, strong B1 fields also label semi-solid magnetization transfer (MT) effects originating from immobile protons with broad linewidths (~kHz) in the tissue. Recently, it was shown that endogenous CEST contrasts are strongly affected by the MT background as well as by T1 relaxation of the water protons. We show that this influence can be corrected in the acquired CEST data by an inverse metric that yields the apparent exchange-dependent relaxation (AREX). AREX has some useful linearity features that enable preparation of both concentration, and--by using the AREX-ratio of two RF irradiation amplitudes B1--purely exchange-rate-weighted CEST contrasts. These two methods could be verified in phantom experiments with different concentration and pH values, but also varying water relaxation properties. Finally, results from a preliminary application to in vivo CEST imaging data of the human calf muscle before and after exercise are presented. The creatine concentration increases during exercise as expected and as confirmed by (31)P NMR spectroscopic imaging. However, the estimated concentrations obtained by our method were higher than the literature values: cCr,rest=24.5±3.74mM to cCr,ex=38.32±13.05mM. The CEST-based pH method shows a pH decrease during exercise, whereas a slight increase was observed by (31)P NMR spectroscopy.

  2. Targeted and Untargeted Metabolomics to Explore the Bioavailability of the Secoiridoids from a Seed/Fruit Extract (Fraxinus angustifolia Vahl) in Human Healthy Volunteers: A Preliminary Study.

    PubMed

    García-Villalba, Rocío; Tomás-Barberán, Francisco A; Fança-Berthon, Pascale; Roller, Marc; Zafrilla, Pilar; Issaly, Nicolas; García-Conesa, María-Teresa

    2015-12-11

    The bark, seeds, fruits and leaves of the genus Fraxinus (Oleaceae) which contain a wide range of phytochemicals, mostly secoiridoid glucosides, have been widely used in folk medicine against a number of ailments, yet little is known about the metabolism and uptake of the major Fraxinus components. The aim of this work was to advance in the knowledge on the bioavailability of the secoiridoids present in a Fraxinus angustifolia Vahl seed/fruit extract using both targeted and untargeted metabolomic analyses. Plasma and urine samples from nine healthy volunteers were taken at specific time intervals following the intake of the extract and analyzed by UPLC-ESI-QTOF. Predicted metabolites such as tyrosol and ligstroside-aglycone glucuronides and sulfates were detected at low intensity. These compounds reached peak plasma levels 2 h after the intake and exhibited high variability among the participants. The ligstroside-aglycone conjugates may be considered as potential biomarkers of the Fraxinus secoiridoids intake. Using the untargeted approach we additionally detected phenolic conjugates identified as ferulic acid and caffeic acid sulfates, as well as hydroxybenzyl and hydroxyphenylacetaldehyde sulfate derivatives which support further metabolism of the secoiridoids by phase I and (or) microbial enzymes. Overall, the results of this study suggest low uptake of intact secoiridoids from a Fraxinus angustifolia Vahl extract in healthy human volunteers and metabolic conversion by esterases, glycosidases, and phase II sulfo- and glucuronosyl transferases to form smaller conjugated derivatives.

  3. Preliminary Study to Test the Feasibility of Sex Identification of Human (Homo sapiens) Bones Based on Differences in Elemental Profiles Determined by Handheld X-ray Fluorescence.

    PubMed

    Nganvongpanit, Korakot; Buddhachat, Kittisak; Brown, Janine L; Klinhom, Sarisa; Pitakarnnop, Tanita; Mahakkanukrauh, Pasuk

    2016-09-01

    Sex assignment of human remains is a crucial step in forensic anthropological studies. The aim of this study was to examine elemental differences between male and female bones using X-ray fluorescence (XRF) and determine if elemental profiling could be used for sex discrimination. Cranium, humerus, and os coxae of 60 skeletons (30 male, 30 female) from the Chiang Mai University Skeletal Collection were scanned by XRF and differences in elemental profiles between male and female bones determined using discriminant analysis. In the cranium, three elements (S, Ca, Pb) were significantly higher in males and five elements (Si, Mn, Fe, Zn, Ag) plus light elements (atomic number lower than 12) were higher in females. In humerus and os coxae, nine elements were significantly higher in male and one element was higher in female samples. The accuracy rate for sex estimation was 60, 63, and 61 % for cranium, humerus, and os coxae, respectively, and 67 % when data for all three bones were combined. We conclude that there are sex differences in bone elemental profiles; however, the accuracy of XRF analyses for discriminating between male and female samples was low compared to standard morphometric and molecular methods. XRF could be used on small samples that cannot be sexed by traditional morphological methods, but more work is needed to increase the power of this technique for gender assignment.

  4. Histological and Histomorphometric Human Results of HA-Beta-TCP 30/70 Compared to Three Different Biomaterials in Maxillary Sinus Augmentation at 6 Months: A Preliminary Report

    PubMed Central

    Annibali, Susanna; Iezzi, Giovanna; Sfasciotti, Gian Luca; Cristalli, Maria Paola; Vozza, Iole; Mangano, Carlo; La Monaca, Gerardo; Polimeni, Antonella

    2015-01-01

    Objective. The aim of this investigation was to examine the bone regenerative potential of newly biphasic calcium phosphate ceramics (HA-β-TCP 30/70), by assessing histological and histomorphometric results of human specimens retrieved from sinuses augmented with HA-β-TCP 30/70, and comparing them to anorganic bovine bone (ABB), mineralized solvent-dehydrated bone allograft (MSDBA), and equine bone (EB), after a healing period of 6 months. Materials and Methods. Four consecutive patients with edentulous atrophic posterior maxilla were included in this report. A two-stage procedure was carried out for sinus augmentation with HA-β-TCP 30/70, ABB, MSDBA, and EB. After 6 months, specimens were retrieved at the time of implant placement and processed for histological and histomorphometric analyses. Results. At histological examination, all biomaterials were in close contact with the newly formed bone and showed the same pattern of bone formation; the grafted granules were surrounded by a bridge-like network of newly formed bone. A limited number of ABB particles were partially covered by connective tissue. The histomorphometric analysis revealed 30.2% newly formed bone for Ha-β-TCP 30/70, 20.1% for ABB, 16.4% for MSDBA, and 21.9% for EB. Conclusions. Within the limitations of the present investigation, these results support the successful use of HA-β-TCP 30/70 for sinus augmentation. PMID:26273589

  5. Preliminary design of a 15 m diameter mechanically scanned deployable offset antenna

    NASA Technical Reports Server (NTRS)

    1983-01-01

    The preliminary design of a 15 meter diameter mechanically scanned, offset rotating, fed parabolic reflector antenna system is reported and the results of preliminary performance, structural and thermal analyses are presented.

  6. Roadway-powered-electric-vehicle impact study: preliminary supporting analyses

    SciTech Connect

    Latorre, V.R.; Spogen, L.R. Jr.

    1981-12-15

    The Roadway-Powered Electric Vehicle (RPEV) impact analysis program is described, and the results to date are presented. Expressions for the number of RPEVs and miles of powered roadways required to accomplish the desired replacement VMT in a USA have been derived, as well as a procedure to determine hourly energy demand resulting from using RPEVs and powered roadways. Conversions factors from VMT to electric energy have also been determined. The input and output data requirements for RECAPS have been determined, as well as the formats for the data for both the EPRI-supported effort, and the simpler, non-EPRI-supported program. The differences between the two programs are also discussed, and data are presented on the average VMT in each major utility service area within the US.

  7. L Chondrite meteorites: A compilation and preliminary analyses

    NASA Technical Reports Server (NTRS)

    Silliman, A.

    1984-01-01

    A compilation of those meteorites currently recognized as being L chondrites, exclusive of the numerous Antarctica finds, was made and is known as the L Chondrite Register. Data for these 576 meteorites was collected from a variety of sources, primarily the British Museum's Catalogue of Meteorites and the Appendix to the Catalogue of Meteorites. Also used was the Revised Cambridge Chondrite Compendium, which provided a convenient listing of L chondrites; other sources include Chinese Meteorites, Meteorites, by Wasson (1974), and the Meteoritical Bulletin of Meteoritics. This last source provided data for most recent falls and was referenced through March of 1982. All such data were recorded on a computer data file with a HP 2647A terminal, so that information could easily be retrieved and manipulated. For each meteorite, the petrographic class, location of find, fall date and hour, mass, mole per cent fayalite, weight per cent Fe, SiO2/MgO ratio, shock class, metal class, 4He abundance, UTh-H3 gas retention age, K-Ar gas retention age, and 21Ne cosmic ray exposure age, was recorded when known.

  8. Preliminary Multiphysics Analyses of HFIR LEU Fuel Conversion using COMSOL

    SciTech Connect

    Freels, James D; Bodey, Isaac T; Arimilli, Rao V; Curtis, Franklin G; Ekici, Kivanc; Jain, Prashant K

    2011-06-01

    The research documented herein was performed by several individuals across multiple organizations. We have previously acknowledged our funding for the project, but another common thread among the authors of this document, and hence the research performed, is the analysis tool COMSOL. The research has been divided into categories to allow the COMSOL analysis to be performed independently to the extent possible. As will be seen herein, the research has progressed to the point where it is expected that next year (2011) a large fraction of the research will require collaboration of our efforts as we progress almost exclusively into three-dimensional (3D) analysis. To the extent possible, we have tried to segregate the development effort into two-dimensional (2D) analysis in order to arrive at techniques and methodology that can be extended to 3D models in a timely manner. The Research Reactors Division (RRD) of ORNL has contracted with the University of Tennessee, Knoxville (UTK) Mechanical, Aerospace and Biomedical Engineering Department (MABE) to perform a significant fraction of this research. This group has been chosen due to their expertise and long-term commitment in using COMSOL and also because the participating students are able to work onsite on a part-time basis due to the close proximity of UTK with the ORNL campus. The UTK research has been governed by a statement of work (SOW) which clearly defines the specific tasks reported herein on the perspective areas of research. Ph.D. student Isaac T. Bodey has focused on heat transfer, fluid flow, modeling, and meshing issues and has been aided by his major professor Dr. Rao V. Arimilli and is the primary contributor to Section 2 of this report. Ph.D student Franklin G. Curtis has been focusing exclusively on fluid-structure interaction (FSI) due to the mechanical forces acting on the plate caused by the flow and has also been aided by his major professor Dr. Kivanc Ekici and is the primary contributor to Section 4 of this report. The HFIR LEU conversion project has also obtained the services of Dr. Prashant K. Jain of the Reactor & Nuclear Systems Division (RNSD) of ORNL. Prashant has quickly adapted to the COMSOL tools and has been focusing on thermal-structure interaction (TSI) issues and development of alternative 3D model approaches that could yield faster-running solutions. Prashant is the primary contributor to Section 5 of the report. And finally, while incorporating findings from all members of the COMSOL team (i.e., the team) and contributing as the senior COMSOL leader and advocate, Dr. James D. Freels has focused on the 3D model development, cluster deployment, and has contributed primarily to Section 3 and overall integration of this report. The team has migrated to the current release of COMSOL at version 4.1 for all the work described in this report, except where stated otherwise. Just as in the performance of the research, each of the respective sections has been originally authored by the respective authors. Therefore, the reader will observe a contrast in writing style throughout this document.

  9. Preliminary Analyses of Beidou Signal-In Anomaly Since 2013

    NASA Astrophysics Data System (ADS)

    Wu, Y.; Ren, J.; Liu, W.

    2016-06-01

    As BeiDou navigation system has been operational since December 2012. There is an increasing desire to use multiple constellation to improve positioning performance. The signal-in-space (SIS) anomaly caused by the ground control and the space vehicle is one of the major threats to affect the integrity. For a young Global Navigation Satellite System, knowledge about SIS anomalies in history is very important for not only assessing the SIS integrity performance of a constellation but also providing the assumption for ARAIM (Advanced Receiver Autonomous Integrity Monitoring). In this paper, the broadcast ephemerides and the precise ones are pre-processed for avoiding the false anomaly identification. The SIS errors over the period of Mar. 2013-Feb. 2016 are computed by comparing the broadcast ephemerides with the precise ones. The time offsets between GPST (GPS time) and BDT (BeiDou time) are estimated and removed by an improved estimation algorithm. SIS worst-UREs are computed and a RMS criteria are investigated to identify the SIS anomalies. The results show that the probability of BeiDou SIS anomalies is in 10-3 level in last three years. Even though BeiDou SIS integrity performance currently cannot match the GPS integrity performances, the result indicates that BeiDou has a tendency to improve its integrity performance.

  10. Comparative Gene Expression Analyses Identify Luminal and Basal Subtypes of Canine Invasive Urothelial Carcinoma That Mimic Patterns in Human Invasive Bladder Cancer.

    PubMed

    Dhawan, Deepika; Paoloni, Melissa; Shukradas, Shweta; Choudhury, Dipanwita Roy; Craig, Bruce A; Ramos-Vara, José A; Hahn, Noah; Bonney, Patty L; Khanna, Chand; Knapp, Deborah W

    2015-01-01

    More than 160,000 people are expected to die from invasive urothelial carcinoma (iUC) this year worldwide. Research in relevant animal models is essential to improving iUC management. Naturally-occurring canine iUC closely resembles human iUC in histopathology, metastatic behavior, and treatment response, and could provide a relevant model for human iUC. The molecular characterization of canine iUC, however, has been limited. Work was conducted to compare gene expression array results between tissue samples from iUC and normal bladder in dogs, with comparison to similar expression array data from human iUC and normal bladder in the literature. Considerable similarities between enrichment patterns of genes in canine and human iUC were observed. These included patterns mirroring basal and luminal subtypes initially observed in human breast cancer and more recently noted in human iUC. Canine iUC samples also exhibited enrichment for genes involved in P53 pathways, as has been reported in human iUC. This is particularly relevant as drugs targeting these genes/pathways in other cancers could be repurposed to treat iUC, with dogs providing a model to optimize therapy. As part of the validation of the results and proof of principal for evaluating individualized targeted therapy, the overexpression of EGFR in canine bladder iUC was confirmed. The similarities in gene expression patterns between dogs and humans add considerably to the value of naturally-occurring canine iUC as a relevant and much needed animal model for human iUC. Furthermore, the finding of expression patterns that cross different pathologically-defined cancers could allow studies of dogs with iUC to help optimize cancer management across multiple cancer types. The work is also expected to lead to a better understanding of the biological importance of the gene expression patterns, and the potential application of the cross-species comparisons approach to other cancer types as well.

  11. Microfluidic affinity and ChIP-seq analyses converge on a conserved FOXP2-binding motif in chimp and human, which enables the detection of evolutionarily novel targets.

    PubMed

    Nelson, Christopher S; Fuller, Chris K; Fordyce, Polly M; Greninger, Alexander L; Li, Hao; DeRisi, Joseph L

    2013-07-01

    The transcription factor forkhead box P2 (FOXP2) is believed to be important in the evolution of human speech. A mutation in its DNA-binding domain causes severe speech impairment. Humans have acquired two coding changes relative to the conserved mammalian sequence. Despite intense interest in FOXP2, it has remained an open question whether the human protein's DNA-binding specificity and chromatin localization are conserved. Previous in vitro and ChIP-chip studies have provided conflicting consensus sequences for the FOXP2-binding site. Using MITOMI 2.0 microfluidic affinity assays, we describe the binding site of FOXP2 and its affinity profile in base-specific detail for all substitutions of the strongest binding site. We find that human and chimp FOXP2 have similar binding sites that are distinct from previously suggested consensus binding sites. Additionally, through analysis of FOXP2 ChIP-seq data from cultured neurons, we find strong overrepresentation of a motif that matches our in vitro results and identifies a set of genes with FOXP2 binding sites. The FOXP2-binding sites tend to be conserved, yet we identified 38 instances of evolutionarily novel sites in humans. Combined, these data present a comprehensive portrait of FOXP2's-binding properties and imply that although its sequence specificity has been conserved, some of its genomic binding sites are newly evolved.

  12. Genome analyses suggest the presence of polyploidy and recent human-driven expansions in eight global populations of the honeybee pathogen Nosema ceranae.

    PubMed

    Pelin, Adrian; Selman, Mohammed; Aris-Brosou, Stéphane; Farinelli, Laurent; Corradi, Nicolas

    2015-11-01

    Nosema ceranae is a microsporidian pathogen whose infections have been associated with recent global declines in the populations of western honeybees (Apis mellifera). Despite the outstanding economic and ecological threat that N. ceranae may represent for honeybees worldwide, many aspects of its biology, including its mode of reproduction, propagation and ploidy, are either very unclear or unknown. In the present study, we set to gain knowledge in these biological aspects by re-sequencing the genome of eight isolates (i.e. a population of spores isolated from one single beehive) of this species harvested from eight geographically distant beehives, and by investigating their level of polymorphism. Consistent with previous analyses performed using single gene sequences, our analyses uncovered the presence of very high genetic diversity within each isolate, but also very little hive-specific polymorphism. Surprisingly, the nature, location and distribution of this genetic variation suggest that beehives around the globe are infected by a population of N. ceranae cells that may be polyploid (4n or more), and possibly clonal. Lastly, phylogenetic analyses based on genome-wide single-nucleotide polymorphism data extracted from these parasites and mitochondrial sequences from their hosts all failed to support the current geographical structure of our isolates.

  13. Diverse approaches to analysing the history of human and pathogen evolution: how to tell the story of the past 70 000 years

    PubMed Central

    Altmann, D. M.; Balloux, F.; Boyton, R. J.

    2012-01-01

    The meeting ‘Human evolution, migration and history revealed by genetics, immunity and infection’, along with the follow-on satellite meeting at the Kavli Centre over the subsequent two days, brought together diverse talents. The aim was to see if new insights could be gained by bringing together those who have interests in the past 50–100 000 years of human history, overlaying the perspectives of palaeogeneticists, anthropologists, human geneticists, pathogen geneticists, immunologists, disease modellers, linguists, immunogeneticists, historians and archaeologists. It rapidly became clear that while all may agree on the broad brush-strokes including ‘out-of-Africa’ and the general approximations of timelines, diverse approaches may often suggest somewhat different ways of telling the story. PMID:22312043

  14. Inter- and intraspecies phylogenetic analyses reveal extensive X-Y gene conversion in the evolution of gametologous sequences of human sex chromosomes.

    PubMed

    Trombetta, Beniamino; Sellitto, Daniele; Scozzari, Rosaria; Cruciani, Fulvio

    2014-08-01

    It has long been believed that the male-specific region of the human Y chromosome (MSY) is genetically independent from the X chromosome. This idea has been recently dismissed due to the discovery that X-Y gametologous gene conversion may occur. However, the pervasiveness of this molecular process in the evolution of sex chromosomes has yet to be exhaustively analyzed. In this study, we explored how pervasive X-Y gene conversion has been during the evolution of the youngest stratum of the human sex chromosomes. By comparing about 0.5 Mb of human-chimpanzee gametologous sequences, we identified 19 regions in which extensive gene conversion has occurred. From our analysis, two major features of these emerged: 1) Several of them are evolutionarily conserved between the two species and 2) almost all of the 19 hotspots overlap with regions where X-Y crossing-over has been previously reported to be involved in sex reversal. Furthermore, in order to explore the dynamics of X-Y gametologous conversion in recent human evolution, we resequenced these 19 hotspots in 68 widely divergent Y haplogroups and used publicly available single nucleotide polymorphism data for the X chromosome. We found that at least ten hotspots are still active in humans. Hence, the results of the interspecific analysis are consistent with the hypothesis of widespread reticulate evolution within gametologous sequences in the differentiation of hominini sex chromosomes. In turn, intraspecific analysis demonstrates that X-Y gene conversion may modulate human sex-chromosome-sequence evolution to a greater extent than previously thought.

  15. A new robotic endoscope manipulator. A preliminary trial to evaluate the performance of a voice-operated industrial robot and a human assistant in several simulated and real endoscopic operations.

    PubMed

    Vara-Thorbeck, C; Muñoz, V F; Toscano, R; Gomez, J; Fernández, J; Felices, M; Garcia-Cerezo, A

    2001-09-01

    We report our learning experience in simulated and real surgical tasks with a new voice-controlled robotic endoscope manipulator: an industrial robot with the tool-holder arm modified to support the optic and camera. The manipulator control-card programs have been rewritten to meet the needs of endoscopic surgeons. For this preliminary work, systems engineers with an additional monitor monitored, recorded, and compared the percentage effectiveness and precision of the responses of the robotic and human assistant to successive oral commands during the several different experimental surgical tasks. Simultaneously, to help develop this voice-commanded system for future, more precise robotic manipulation of surgical instruments, they measured the cartesian and spherical coordinates of successive positions of the optic. In unexpectedly difficult experimental conditions, the tireless robot proved more precise and effective than the demonstrably fatigable human: the steadier screen images of the robotic manipulations helped the surgeon tie knots in 7-0 sutures.

  16. EEG analyses with SOBI.

    SciTech Connect

    Glickman, Matthew R.; Tang, Akaysha

    2009-02-01

    The motivating vision behind Sandia's MENTOR/PAL LDRD project has been that of systems which use real-time psychophysiological data to support and enhance human performance, both individually and of groups. Relevant and significant psychophysiological data being a necessary prerequisite to such systems, this LDRD has focused on identifying and refining such signals. The project has focused in particular on EEG (electroencephalogram) data as a promising candidate signal because it (potentially) provides a broad window on brain activity with relatively low cost and logistical constraints. We report here on two analyses performed on EEG data collected in this project using the SOBI (Second Order Blind Identification) algorithm to identify two independent sources of brain activity: one in the frontal lobe and one in the occipital. The first study looks at directional influences between the two components, while the second study looks at inferring gender based upon the frontal component.

  17. Paleoecological analyses of lake sediments reveal prehistoric human impact on forests at Anthony Island UNESCO World Heritage Site, Queen Charlotte Islands (Haida Gwaii), Canada

    NASA Astrophysics Data System (ADS)

    Lacourse, Terri; Mathewes, Rolf W.; Hebda, Richard J.

    2007-09-01

    Pollen and plant macrofossil analyses of lake sediments from Anthony Island in the southern Queen Charlotte Islands (Haida Gwaii), British Columbia, reveal 1800 yr of relatively stable temperate rainforest vegetation. Cupressaceae (cedar) pollen percentages and accumulation rates decline about 1000 cal yr BP, coincident with occupation of the island by Haida peoples, who use Thuja plicata (western red cedar) almost exclusively for house construction, dugout canoes, monumental poles, and many other items. Anthropogenic disturbance offers the most likely explanation for the decline of T. plicata.

  18. Analysis and Modelling for Human Resource Management in Defence (Analyse et modelisation de la gestion des ressources humaines en matiere de defense)

    DTIC Science & Technology

    2009-03-01

    PROJECT NUMBER 5e. TASK NUMBER 5f. WORK UNIT NUMBER 7. PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) 8. PERFORMING ORGANIZATION...RTO performs its mission with the support of an extensive network of national experts. It also ensures effective co-ordination with other NATO bodies... logistique . La réunion des spécialistes du domaine de la recherche SAS-073 en analyse et en modélisation de la gestion des ressources humaines pour la

  19. Mitochondrial DNA and STR analyses for human DNA from maggots crop contents: a forensic entomology case from central-southern China.

    PubMed

    Li, X; Cai, J F; Guo, Y D; Xiong, F; Zhang, L; Feng, H; Meng, F M; Fu, Y; Li, J B; Chen, Y Q

    2011-08-01

    Insect larvae and adult insects found on human corpses can provide important forensic evidence however it is useful to be able to prove evidence of association. Without this, it could be claimed that the insect evidence was a contaminant or had been planted on the body. This paper describes how mitochondrial DNA (mtDNA) and STR analysis of the crop contents of larvae of the blowfly Aldrichina grahami collected from separated body parts was used to provide evidence of association.

  20. Molecular modeling of the human eukaryotic translation initiation factor 5A (eIF5A) based on spectroscopic and computational analyses

    SciTech Connect

    Costa-Neto, Claudio M. . E-mail: claudio@fmrp.usp.br; Parreiras-e-Silva, Lucas T.; Ruller, Roberto; Oliveira, Eduardo B.; Miranda, Antonio; Oliveira, Laerte; Ward, Richard J.

    2006-09-01

    The eukaryotic translation initiation factor 5A (eIF5A) is a protein ubiquitously present in archaea and eukarya, which undergoes a unique two-step post-translational modification called hypusination. Several studies have shown that hypusination is essential for a variety of functional roles for eIF5A, including cell proliferation and synthesis of proteins involved in cell cycle control. Up to now neither a totally selective inhibitor of hypusination nor an inhibitor capable of directly binding to eIF5A has been reported in the literature. The discovery of such an inhibitor might be achieved by computer-aided drug design based on the 3D structure of the human eIF5A. In this study, we present a molecular model for the human eIF5A protein based on the crystal structure of the eIF5A from Leishmania brasiliensis, and compare the modeled conformation of the loop bearing the hypusination site with circular dichroism data obtained with a synthetic peptide of this loop. Furthermore, analysis of amino acid variability between different human eIF5A isoforms revealed peculiar structural characteristics that are of functional relevance.

  1. Does low-energy sweetener consumption affect energy intake and body weight? A systematic review, including meta-analyses, of the evidence from human and animal studies.

    PubMed

    Rogers, P J; Hogenkamp, P S; de Graaf, C; Higgs, S; Lluch, A; Ness, A R; Penfold, C; Perry, R; Putz, P; Yeomans, M R; Mela, D J

    2016-03-01

    By reducing energy density, low-energy sweeteners (LES) might be expected to reduce energy intake (EI) and body weight (BW). To assess the totality of the evidence testing the null hypothesis that LES exposure (versus sugars or unsweetened alternatives) has no effect on EI or BW, we conducted a systematic review of relevant studies in animals and humans consuming LES with ad libitum access to food energy. In 62 of 90 animal studies exposure to LES did not affect or decreased BW. Of 28 reporting increased BW, 19 compared LES with glucose exposure using a specific 'learning' paradigm. Twelve prospective cohort studies in humans reported inconsistent associations between LES use and body mass index (-0.002 kg m(-)(2) per year, 95% confidence interval (CI) -0.009 to 0.005). Meta-analysis of short-term randomized controlled trials (129 comparisons) showed reduced total EI for LES versus sugar-sweetened food or beverage consumption before an ad libitum meal (-94 kcal, 95% CI -122 to -66), with no difference versus water (-2 kcal, 95% CI -30 to 26). This was consistent with EI results from sustained intervention randomized controlled trials (10 comparisons). Meta-analysis of sustained intervention randomized controlled trials (4 weeks to 40 months) showed that consumption of LES versus sugar led to relatively reduced BW (nine comparisons; -1.35 kg, 95% CI -2.28 to -0.42), and a similar relative reduction in BW versus water (three comparisons; -1.24 kg, 95% CI -2.22 to -0.26). Most animal studies did not mimic LES consumption by humans, and reverse causation may influence the results of prospective cohort studies. The preponderance of evidence from all human randomized controlled trials indicates that LES do not increase EI or BW, whether compared with caloric or non-caloric (for example, water) control conditions. Overall, the balance of evidence indicates that use of LES in place of sugar, in children and adults, leads to reduced EI and BW, and possibly also

  2. Broad Blockade Antibody Responses in Human Volunteers after Immunization with a Multivalent Norovirus VLP Candidate Vaccine: Immunological Analyses from a Phase I Clinical Trial

    PubMed Central

    Lindesmith, Lisa C.; Ferris, Martin T.; Mullan, Clancy W.; Ferreira, Jennifer; Debbink, Kari; Swanstrom, Jesica; Richardson, Charles; Goodwin, Robert R.; Baehner, Frank; Mendelman, Paul M.; Bargatze, Robert F.; Baric, Ralph S.

    2015-01-01

    Background Human noroviruses (NoVs) are the primary cause of acute gastroenteritis and are characterized by antigenic variation between genogroups and genotypes and antigenic drift of strains within the predominant GII.4 genotype. In the context of this diversity, an effective NoV vaccine must elicit broadly protective immunity. We used an antibody (Ab) binding blockade assay to measure the potential cross-strain protection provided by a multivalent NoV virus-like particle (VLP) candidate vaccine in human volunteers. Methods and Findings Sera from ten human volunteers immunized with a multivalent NoV VLP vaccine (genotypes GI.1/GII.4) were analyzed for IgG and Ab blockade of VLP interaction with carbohydrate ligand, a potential correlate of protective immunity to NoV infection and illness. Immunization resulted in rapid rises in IgG and blockade Ab titers against both vaccine components and additional VLPs representing diverse strains and genotypes not represented in the vaccine. Importantly, vaccination induced blockade Ab to two novel GII.4 strains not in circulation at the time of vaccination or sample collection. GII.4 cross-reactive blockade Ab titers were more potent than responses against non-GII.4 VLPs, suggesting that previous exposure history to this dominant circulating genotype may impact the vaccine Ab response. Further, antigenic cartography indicated that vaccination preferentially activated preexisting Ab responses to epitopes associated with GII.4.1997. Study interpretations may be limited by the relevance of the surrogate neutralization assay and the number of immunized participants evaluated. Conclusions Vaccination with a multivalent NoV VLP vaccine induces a broadly blocking Ab response to multiple epitopes within vaccine and non-vaccine NoV strains and to novel antigenic variants not yet circulating at the time of vaccination. These data reveal new information about complex NoV immune responses to both natural exposure and to vaccination, and

  3. Does low-energy sweetener consumption affect energy intake and body weight? A systematic review, including meta-analyses, of the evidence from human and animal studies

    PubMed Central

    Rogers, P J; Hogenkamp, P S; de Graaf, C; Higgs, S; Lluch, A; Ness, A R; Penfold, C; Perry, R; Putz, P; Yeomans, M R; Mela, D J

    2016-01-01

    By reducing energy density, low-energy sweeteners (LES) might be expected to reduce energy intake (EI) and body weight (BW). To assess the totality of the evidence testing the null hypothesis that LES exposure (versus sugars or unsweetened alternatives) has no effect on EI or BW, we conducted a systematic review of relevant studies in animals and humans consuming LES with ad libitum access to food energy. In 62 of 90 animal studies exposure to LES did not affect or decreased BW. Of 28 reporting increased BW, 19 compared LES with glucose exposure using a specific ‘learning' paradigm. Twelve prospective cohort studies in humans reported inconsistent associations between LES use and body mass index (−0.002 kg m−2 per year, 95% confidence interval (CI) −0.009 to 0.005). Meta-analysis of short-term randomized controlled trials (129 comparisons) showed reduced total EI for LES versus sugar-sweetened food or beverage consumption before an ad libitum meal (−94 kcal, 95% CI −122 to −66), with no difference versus water (−2 kcal, 95% CI −30 to 26). This was consistent with EI results from sustained intervention randomized controlled trials (10 comparisons). Meta-analysis of sustained intervention randomized controlled trials (4 weeks to 40 months) showed that consumption of LES versus sugar led to relatively reduced BW (nine comparisons; −1.35 kg, 95% CI –2.28 to −0.42), and a similar relative reduction in BW versus water (three comparisons; −1.24 kg, 95% CI –2.22 to −0.26). Most animal studies did not mimic LES consumption by humans, and reverse causation may influence the results of prospective cohort studies. The preponderance of evidence from all human randomized controlled trials indicates that LES do not increase EI or BW, whether compared with caloric or non-caloric (for example, water) control conditions. Overall, the balance of evidence indicates that use of LES in place of sugar, in children and adults, leads to reduced EI

  4. Evaluating mesenchymal stem cell therapy for sepsis with preclinical meta-analyses prior to initiating a first-in-human trial

    PubMed Central

    Lalu, Manoj M; Sullivan, Katrina J; Mei, Shirley HJ; Moher, David; Straus, Alexander; Fergusson, Dean A; Stewart, Duncan J; Jazi, Mazen; MacLeod, Malcolm; Winston, Brent; Marshall, John; Hutton, Brian; Walley, Keith R; McIntyre, Lauralyn

    2016-01-01

    Evaluation of preclinical evidence prior to initiating early-phase clinical studies has typically been performed by selecting individual studies in a non-systematic process that may introduce bias. Thus, in preparation for a first-in-human trial of mesenchymal stromal cells (MSCs) for septic shock, we applied systematic review methodology to evaluate all published preclinical evidence. We identified 20 controlled comparison experiments (980 animals from 18 publications) of in vivo sepsis models. Meta-analysis demonstrated that MSC treatment of preclinical sepsis significantly reduced mortality over a range of experimental conditions (odds ratio 0.27, 95% confidence interval 0.18–0.40, latest timepoint reported for each study). Risk of bias was unclear as few studies described elements such as randomization and no studies included an appropriately calculated sample size. Moreover, the presence of publication bias resulted in a ~30% overestimate of effect and threats to validity limit the strength of our conclusions. This novel prospective application of systematic review methodology serves as a template to evaluate preclinical evidence prior to initiating first-in-human clinical studies. DOI: http://dx.doi.org/10.7554/eLife.17850.001 PMID:27870924

  5. A fast method for analysing six perfluoroalkyl substances in human serum by solid-phase extraction on-line coupled to liquid chromatography tandem mass spectrometry.

    PubMed

    Bartolomé, Mónica; Gallego-Picó, Alejandrina; Huetos, Olga; Lucena, Miguel Ángel; Castaño, Argelia

    2016-03-01

    We have developed and validated an on-line TurboFlow solid-phase extraction procedure coupled to high-performance liquid chromatography with tandem mass spectrometry for the analysis of six perfluoroalkyl substances (PFAS), two sulfonates (perfluorooctane sulfonate and perfluorohexane sulfonate), three carboxylates (perfluorooctanoic acid, perfluorononanoic acid and perfluorodecanoic acid), and one sulfonamide (N-methylperfluorooctane sulfonamide), in human serum samples. This method requires only 100 μL of sample and involves a short pre-treatment with acetonitrile followed by addition of a labelled internal standard for quantification and ultracentrifugation. All PFAS were detected with a run time of 8.5 min. Linearity ranges stay between 0.1 and 20 μg L(-1) (R (2) > 0.9960). Recoveries were determined by spiking blank serum samples with a mixture of six PFAS and found to be in the range 96-110 % for all compounds. Isotopic dilution was used to quantify the selected analytes. The low limits of quantification obtained, between 0.16 and 0.34 μg L(-1), small volume of sample required and short run time used (from two to three times shorter than any other described method), make this validated method highly recommended for human biomonitoring studies.

  6. Anodal transcranial direct current stimulation (tDCS) increases frontal-midline theta activity in the human EEG: a preliminary investigation of non-invasive stimulation.

    PubMed

    Miller, Joe; Berger, Barbara; Sauseng, Paul

    2015-02-19

    Rhythmical brain activity in the range between four and eight Hz acquired over frontal-midline EEG recording sites - so called frontal-midline theta activity - is regarded as one of the most prominent neural signatures of sustained attention. It is reported to parametrically increase with cognitive load and is thought to be generated in medial prefrontal cortex. Here we explored the possibility of using anodal transcranial direct current stimulation over frontal sites to enhance frontal-midline theta activity and to increase sustained attention performance. We used a small preliminary sample to test a novel direct current stimulation electrode configuration by which we were able to significantly increase frontal-midline theta amplitude in a resting condition after the end of the stimulation period. Using standardised low resolution electromagnetic tomography analysis the effect in the surface EEG was localised to right prefrontal and left medial prefrontal brain areas. Transcranial direct current stimulation did, however, not have any impact on behavioural performance during a sustained attention task. This most likely was due to a very fast washout of the stimulation's after effect on theta activity. Although these are only preliminary results from a rather small sample, this study demonstrates that transcranial direct current stimulation can be used to rather selectively enhance frontal-midline theta amplitude.

  7. Risk assessment of PCDD/Fs levels in human tissues related to major food items based on chemical analyses and micro-EROD assay.

    PubMed

    Tsang, H L; Wu, S C; Wong, C K C; Leung, C K M; Tao, S; Wong, M H

    2009-10-01

    Nine groups of food items (freshwater fish, marine fish, pork, chicken, chicken eggs, leafy, non-leafy vegetables, rice and flour) and three types of human samples (human milk, maternal serum and cord serum) were collected for the analysis of PCDD/Fs. Results of chemical analysis revealed PCDD/Fs concentrations (pg g(-1) fat) in the following ascending order: pork (0.289 pg g(-1) fat), grass carp (Ctenopharyngodon idellus) (freshwater fish) (0.407), golden thread (Nemipterus virgatus) (marine fish) (0.511), chicken (0.529), mandarin fish (Siniperca kneri) (marine fish) (0.535), chicken egg (0.552), and snubnose pompano (Trachinotus blochii) (marine fish) (1.219). The results of micro-EROD assay showed relatively higher PCDD/Fs levels in fish (2.65 pg g(-1) fat) when compared with pork (0.47), eggs (0.33), chicken (0.13), flour (0.07), vegetables (0.05 pg g(-1) wet wt) and rice (0.05). The estimated average daily intake of PCDD/Fs of 3.51 pg EROD-TEQ/kg bw/day was within the range of WHO Tolerable Daily Intake (1-4 pg WHO-TEQ/kg bw/day) and was higher than the Provisional Tolerable Daily Intake (PMTL) (70 pg for dioxins and dioxin-like PCBs) recommended by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) [Joint FAO/WHO Expert Committee on Food Additives (JECFA), Summary and conclusions of the fifty-seventh meeting, JECFA, 2001.]. Nevertheless, the current findings were significantly lower than the TDI (14 pg WHO-TEQ/kg/bw/day) recommended by the Scientific Committee on Food of the Europe Commission [European Scientific Committee on Food (EU SCF), Opinions on the SCF on the risk assessment of dioxins and dioxin-like PCBs in food, 2000.]. However, it should be noted that micro-EROD assay overestimates the PCDD/Fs levels by 2 to 7 folds which may also amplify the PCDD/Fs levels accordingly. Although the levels of PCDD/Fs obtained from micro-EROD assay were much higher than those obtained by chemical analysis by 2 to 7 folds, it provides a cost-effective and

  8. Preexisting CD4+ T-cell immunity in human population to avian influenza H7N9 virus: whole proteome-wide immunoinformatics analyses.

    PubMed

    Duvvuri, Venkata R; Duvvuri, Bhargavi; Alice, Christilda; Wu, Gillian E; Gubbay, Jonathan B; Wu, Jianhong

    2014-01-01

    In 2013, a novel avian influenza H7N9 virus was identified in human in China. The antigenically distinct H7N9 surface glycoproteins raised concerns about lack of cross-protective neutralizing antibodies. Epitope-specific preexisting T-cell immunity was one of the protective mechanisms in pandemic 2009 H1N1 even in the absence of cross-protective antibodies. Hence, the assessment of preexisting CD4+ T-cell immunity to conserved epitopes shared between H7N9 and human influenza A viruses (IAV) is critical. A comparative whole proteome-wide immunoinformatics analysis was performed to predict the CD4+ T-cell epitopes that are commonly conserved within the proteome of H7N9 in reference to IAV subtypes (H1N1, H2N2, and H3N2). The CD4+ T-cell epitopes that are commonly conserved (∼ 556) were further screened against the Immune Epitope Database (IEDB) to validate their immunogenic potential. This analysis revealed that 45.5% (253 of 556) epitopes are experimentally proven to induce CD4+ T-cell memory responses. In addition, we also found that 23.3% of CD4+ T-cell epitopes have ≥ 90% of sequence homology with experimentally defined CD8+ T-cell epitopes. We also conducted the population coverage analysis across different ethnicities using commonly conserved CD4+ T-cell epitopes and corresponding HLA-DRB1 alleles. Interestingly, the indigenous populations from Canada, United States, Mexico and Australia exhibited low coverage (28.65% to 45.62%) when compared with other ethnicities (57.77% to 94.84%). In summary, the present analysis demonstrate an evidence on the likely presence of preexisting T-cell immunity in human population and also shed light to understand the potential risk of H7N9 virus among indigenous populations, given their high susceptibility during previous pandemic influenza events. This information is crucial for public health policy, in targeting priority groups for immunization programs.

  9. Analyses of Resected Human Brain Metastases of Breast Cancer Reveal the Association between Up-regulation of Hexokinase 2 and Poor Prognosis

    PubMed Central

    Palmieri, Diane; Fitzgerald, Daniel; Shreeve, S. Martin; Hua, Emily; Bronder, Julie L.; Weil, Robert J.; Davis, Sean; Stark, Andreas M.; Merino, Maria J.; Kurek, Raffael; Mehdorn, H. Maximilian; Davis, Gary; Steinberg, Seth M.; Meltzer, Paul S.; Aldape, Kenneth; Steeg, Patricia S.

    2009-01-01

    Brain metastases of breast cancer appear to be increasing in incidence as systemic therapy improves. Metastatic disease in the brain is associated with high morbidity and mortality. We present the first gene expression analysis of laser captured epithelial cells from resected human brain metastases of breast cancer compared to unlinked primary breast tumors. The tumors were matched for histology, TNM stage and hormone receptor status. Most differentially expressed genes were down-regulated in the brain metastases which included, surprisingly, many genes associated with metastasis. Q-PCR analysis confirmed statistically significant differences or strong trends in the expression of six genes: BMP1, PEDF, LAMγ3, SIAH, STHMN3 and TSPD2. Hexokinase 2 (HK2) was also of interest because of its increased expression in brain metastases. HK2 is important in glucose metabolism and apoptosis. In agreement with our microarray results, HK2 levels (both mRNA and protein) were elevated in a brain metastatic derivative (231-BR) of the human breast carcinoma cell line MDA-MB-231 relative to the parental cell line (231-P), in vitro. Knockdown of HK2 expression in 231-BR cells using shRNA reduced cell proliferation when cultures were maintained in glucose limiting conditions. Finally, HK2 expression was analyzed in a cohort of 123 resected brain metastases of breast cancer. High HK2 expression was significantly associated with poor patient survival post-craniotomy (P=0.028). The data suggest that HK2 overexpression is associated with metastasis to the brain in breast cancer and it may be a therapeutic target. PMID:19723875

  10. A Luciferase Gene Driven by an Alphaherpesviral Promoter Also Responds to Immediate Early Antigens of the Betaherpesvirus HCMV, Allowing Comparative Analyses of Different Human Herpesviruses in One Reporter Cell Line

    PubMed Central

    Villinger, Clarissa; Schubert, Axel; Walther, Paul; Sinzger, Christian; Lieber, Diana

    2017-01-01

    Widely used methods for quantification of human cytomegalovirus (HCMV) infection in cell culture such as immunoblotting or plaque reduction assays are generally restricted to low throughput and require time-consuming evaluation. Up to now, only few HCMV reporter cell lines have been generated to overcome these restrictions and they are afflicted with other limitations because permanently expandable cell lines are normally not fully permissive to HCMV. In this work, a previously existing epithelial cell line hosting a luciferase gene under control of a Varicella-zoster virus promoter was adopted to investigate HCMV infection. The cells were susceptible to different HCMV strains at infection efficiencies that corresponded to their respective degree of epithelial cell tropism. Expression of early and late viral antigens, formation of nuclear inclusions, release of infectious virus progeny, and focal growth indicated productive viral replication. However, viral release and spread occurred at lower levels than in primary cell lines which appears to be due to a malfunction of virion morphogenesis during the nuclear stage. Expression of the luciferase reporter gene was specifically induced in HCMV infected cultures as a function of the virus dose and dependent on viral immediate early gene expression. The level of reporter activity accurately reflected infection efficiencies as determined by viral antigen immunostaining, and hence could discriminate the cell tropism of the tested virus strains. As proof-of-principle, we demonstrate that this cell line is applicable to evaluate drug resistance of clinical HCMV isolates and the neutralization capacity of human sera, and that it allows comparative and simultaneous analysis of HCMV and human herpes simplex virus type 1. In summary, the permanent epithelial reporter cell line allows robust, rapid and objective quantitation of HCMV infection and it will be particularly useful in higher throughput analyses as well as in

  11. Quantitative analyses of the effect of silk fibroin/nano-hydroxyapatite composites on osteogenic differentiation of MG-63 human osteosarcoma cells.

    PubMed

    Lin, Linxue; Hao, Runsong; Xiong, Wei; Zhong, Jian

    2015-05-01

    Silk fibroin (SF)/nano-hydroxyapatite (n-HA) composites are potential biomaterials for bone defect repair. Up to now, the biological evaluation studies of SF/n-HA composites have primarily concentrated on their biocompatibility at cell level such as cell viability and proliferation and tissue level such as material absorption and new bone formation. In this work, SF/n-HA composites were fabricated using a simplified coprecipitation methods and were deposited onto Ti alloy substrates. Then the cell adhesion ability of SF/n-HA composites was observed by SEM and cell proliferation ability of SF/n-HA composites was determined by MTT assay. The ALP activity, BGP contents, and Col I contents of MG-63 human osteosarcoma cells on SF/n-HA composites were quantitatively analyzed. HA nanocrystals were used as controls. These experiments showed that SF/n-HA composites had better cell adhesion and osteogenic differentiation abilities than n-HA materials. This work provides quantitative data to analyze the effect of SF/n-HA composites on cell osteogenic differentiation.

  12. Molecular Comparison and Evolutionary Analyses of VP1 Nucleotide Sequences of New African Human Enterovirus 71 Isolates Reveal a Wide Genetic Diversity

    PubMed Central

    Nougairède, Antoine; Joffret, Marie-Line; Deshpande, Jagadish M.; Dubot-Pérès, Audrey; Héraud, Jean-Michel

    2014-01-01

    Most circulating strains of Human enterovirus 71 (EV-A71) have been classified primarily into three genogroups (A to C) on the basis of genetic divergence between the 1D gene, which encodes the VP1 capsid protein. The aim of the present study was to provide further insights into the diversity of the EV-A71 genogroups following the recent description of highly divergent isolates, in particular those from African countries, including Madagascar. We classified recent EV-A71 isolates by a large comparison of 3,346 VP1 nucleotidic sequences collected from GenBank. Analysis of genetic distances and phylogenetic investigations indicated that some recently-reported isolates did not fall into the genogroups A-C and clustered into three additional genogroups, including one Indian genogroup (genogroup D) and 2 African ones (E and F). Our Bayesian phylogenetic analysis provided consistent data showing that the genogroup D isolates share a recent common ancestor with the members of genogroup E, while the isolates of genogroup F evolved from a recent common ancestor shared with the members of the genogroup B. Our results reveal the wide diversity that exists among EV-A71 isolates and suggest that the number of circulating genogroups is probably underestimated, particularly in developing countries where EV-A71 epidemiology has been poorly studied. PMID:24598878

  13. Genetic, Structural, and Antigenic Analyses of Glycan Diversity in the O-Linked Protein Glycosylation Systems of Human Neisseria Species▿ †

    PubMed Central

    Børud, Bente; Aas, Finn Erik; Vik, Åshild; Winther-Larsen, Hanne C.; Egge-Jacobsen, Wolfgang; Koomey, Michael

    2010-01-01

    Bacterial capsular polysaccharides and lipopolysaccharides are well-established ligands of innate and adaptive immune effectors and often exhibit structural and antigenic variability. Although many surface-localized glycoproteins have been identified in bacterial pathogens and symbionts, it not clear if and how selection impacts associated glycoform structure. Here, a systematic approach was devised to correlate gene repertoire with protein-associated glycoform structure in Neisseria species important to human health and disease. By manipulating the protein glycosylation (pgl) gene content and assessing the glycan structure by mass spectrometry and reactivity with monoclonal antibodies, it was established that protein-associated glycans are antigenically variable and that at least nine distinct glycoforms can be expressed in vitro. These studies also revealed that in addition to Neisseria gonorrhoeae strain N400, one other gonococcal strain and isolates of Neisseria meningitidis and Neisseria lactamica exhibit broad-spectrum O-linked protein glycosylation. Although a strong correlation between pgl gene content, glycoform expression, and serological profile was observed, there were significant exceptions, particularly with regard to levels of microheterogeneity. This work provides a technological platform for molecular serotyping of neisserial protein glycans and for elucidating pgl gene evolution. PMID:20363948

  14. Preliminary ECLSS waste water model

    NASA Technical Reports Server (NTRS)

    Carter, Donald L.; Holder, Donald W., Jr.; Alexander, Kevin; Shaw, R. G.; Hayase, John K.

    1991-01-01

    A preliminary waste water model for input to the Space Station Freedom (SSF) Environmental Control and Life Support System (ECLSS) Water Processor (WP) has been generated for design purposes. Data have been compiled from various ECLSS tests and flight sample analyses. A discussion of the characterization of the waste streams comprising the model is presented, along with a discussion of the waste water model and the rationale for the inclusion of contaminants in their respective concentrations. The major objective is to establish a methodology for the development of a waste water model and to present the current state of that model.

  15. Crystallization and preliminary X-ray diffraction studies of a surface mutant of the middle domain of PB2 from human influenza A (H1N1) virus.

    PubMed

    Tsurumura, Toshiharu; Qiu, Hao; Yoshida, Toru; Tsumori, Yayoi; Tsuge, Hideaki

    2014-01-01

    In the last hundred years, four influenza pandemics have been experienced, beginning with that in Spain in 1918. Influenza A virus causes severe pneumonia and its RNA polymerase is an important target for drug design. The influenza A (H1N1) virus has eight ribonucleoprotein complexes, which are composed of viral RNA, RNA polymerases and nucleoproteins. PB2 forms part of the RNA polymerase complex and plays an important role in binding to the cap structure of host mRNA. The middle domain of PB2 includes a cap-binding site. The structure of PB2 from H1N1 complexed with m(7)GTP has not been reported. Plate-like crystals of the middle domain of PB2 from H1N1 were obtained, but the quality of these crystals was not good. An attempt was made to crystallize the middle domain of PB2 complexed with m(7)GTP using a soaking method; however, electron density for m(7)GTP was not observed on preliminary X-ray diffraction analysis. This protein has hydrophobic residues on its surface and is stable in the presence of high salt concentrations. To improve the solubility, a surface double mutant (P453H and I471T) was prepared. These mutations change the surface electrostatic potential drastically. The protein was successfully prepared at a lower salt concentration and good cube-shaped crystals were obtained using this protein. Here, the crystallization and preliminary X-ray diffraction analysis of this mutant of the middle domain of PB2 are reported.

  16. Teleoperator Human Factors Study

    NASA Technical Reports Server (NTRS)

    1986-01-01

    An investigation of the spectrum of space teleoperation activities likely in the 1985 to 1995 decade focused on the resolution of critical human engineering issues and characterization of the technology effect on performance of remote human operators. The study began with the identification and documentation of a set of representative reference teleoperator tasks. For each task, technology, development, and design options, issues, and alternatives that bear on human operator performance were defined and categorized. A literature survey identified existing studies of man/machine issues. For each teleoperations category, an assessment was made of the state of knowledge on a scale from adequate to void. The tests, experiments, and analyses necessary to provide the missing elements of knowledge were then defined. A limited set of tests were actually performed, including operator selection, baseline task definition, control mode study, lighting study, camera study, and preliminary time delay study.

  17. Finite element analyses of human vertebral bodies embedded in polymethylmethalcrylate or loaded via the hyperelastic intervertebral disc models provide equivalent predictions of experimental strength.

    PubMed

    Lu, Yongtao; Maquer, Ghislain; Museyko, Oleg; Püschel, Klaus; Engelke, Klaus; Zysset, Philippe; Morlock, Michael; Huber, Gerd

    2014-07-18

    Quantitative computer tomography (QCT)-based finite element (FE) models of vertebral body provide better prediction of vertebral strength than dual energy X-ray absorptiometry. However, most models were validated against compression of vertebral bodies with endplates embedded in polymethylmethalcrylate (PMMA). Yet, loading being as important as bone density, the absence of intervertebral disc (IVD) affects the strength. Accordingly, the aim was to assess the strength predictions of the classic FE models (vertebral body embedded) against the in vitro and in silico strengths of vertebral bodies loaded via IVDs. High resolution peripheral QCT (HR-pQCT) were performed on 13 segments (T11/T12/L1). T11 and L1 were augmented with PMMA and the samples were tested under a 4° wedge compression until failure of T12. Specimen-specific model was generated for each T12 from the HR-pQCT data. Two FE sets were created: FE-PMMA refers to the classical vertebral body embedded model under axial compression; FE-IVD to their loading via hyperelastic IVD model under the wedge compression as conducted experimentally. Results showed that FE-PMMA models overestimated the experimental strength and their strength prediction was satisfactory considering the different experimental set-up. On the other hand, the FE-IVD models did not prove significantly better (Exp/FE-PMMA: R²=0.68; Exp/FE-IVD: R²=0.71, p=0.84). In conclusion, FE-PMMA correlates well with in vitro strength of human vertebral bodies loaded via real IVDs and FE-IVD with hyperelastic IVDs do not significantly improve this correlation. Therefore, it seems not worth adding the IVDs to vertebral body models until fully validated patient-specific IVD models become available.

  18. History of human activity in last 800 years reconstructed from combined archive data and high-resolution analyses of varved lake sediments from Lake Czechowskie, Northern Poland

    NASA Astrophysics Data System (ADS)

    Słowiński, Michał; Tyszkowski, Sebastian; Ott, Florian; Obremska, Milena; Kaczmarek, Halina; Theuerkauf, Martin; Wulf, Sabine; Brauer, Achim

    2016-04-01

    The aim of the study was to reconstruct human and landscape development in the Tuchola Pinewoods (Northern Poland) during the last 800 years. We apply an approach that combines historic maps and documents with pollen data. Pollen data were obtained from varved lake sediments at a resolution of 5 years. The chronology of the sediment record is based on varve counting, AMS 14C dating, 137Cs activity concentration measurements and tephrochronology (Askja AD 1875). We applied the REVEALS model to translate pollen percentage data into regional plant abundances. The interpretation of the pollen record is furthermore based on pollen accumulation rate data. The pollen record and historic documents show similar trends in vegetation development. During the first phase (AD 1200-1412), the Lake Czechowskie area was still largely forested with Quercus, Carpinus and Pinus forests. Vegetation was more open during the second phase (AD 1412-1776), and reached maximum openness during the third phase (AD 1776-1905). Furthermore, intensified forest management led to a transformation from mixed to pine dominated forests during this period. Since the early 20th century, the forest cover increased again with dominance of the Scots pine in the stand. While pollen and historic data show similar trends, they differ substantially in the degree of openness during the four phases with pollen data commonly suggesting more open conditions. We discuss potential causes for this discrepancy, which include unsuitable parameters settings in REVEALS and unknown changes in forest structure. Using pollen accumulation data as a third proxy record we aim to identify the most probable causes. Finally, we discuss the observed vegetation change in relation the socio-economic development of the area. This study is a contribution to the Virtual Institute of Integrated Climate and Landscape Evolution Analysis - ICLEA- of the Helmholtz Association and National Science Centre, Poland (grant No. 2011/01/B/ST10

  19. High-Resolution Analyses of Human Leukocyte Antigens Allele and Haplotype Frequencies Based on 169,995 Volunteers from the China Bone Marrow Donor Registry Program

    PubMed Central

    Zhou, Xiao-Yang; Zhu, Fa-Ming; Li, Jian-Ping; Mao, Wei; Zhang, De-Mei; Liu, Meng-Li; Hei, Ai-Lian; Dai, Da-Peng; Jiang, Ping; Shan, Xiao-Yan; Zhang, Bo-Wei; Zhu, Chuan-Fu; Shen, Jie; Deng, Zhi-Hui; Wang, Zheng-Lei; Yu, Wei-Jian; Chen, Qiang; Qiao, Yan-Hui; Zhu, Xiang-Ming; Lv, Rong; Li, Guo-Ying; Li, Guo-Liang; Li, Heng-Cong; Zhang, Xu; Pei, Bin; Jiao, Li-Xin; Shen, Gang; Liu, Ying; Feng, Zhi-Hui; Su, Yu-Ping; Xu, Zhao-Xia; Di, Wen-Ying; Jiang, Yao-Qin; Fu, Hong-Lei; Liu, Xiang-Jun; Liu, Xiang; Zhou, Mei-Zhen; Du, Dan; Liu, Qi; Han, Ying; Zhang, Zhi-Xin; Cai, Jian-Ping

    2015-01-01

    Allogeneic hematopoietic stem cell transplantation is a widely used and effective therapy for hematopoietic malignant diseases and numerous other disorders. High-resolution human leukocyte antigen (HLA) haplotype frequency distributions not only facilitate individual donor searches but also determine the probability with which a particular patient can find HLA-matched donors in a registry. The frequencies of the HLA-A, -B, -C, -DRB1, and -DQB1 alleles and haplotypes were estimated among 169,995 Chinese volunteers using the sequencing-based typing (SBT) method. Totals of 191 HLA-A, 244 HLA-B, 146 HLA-C, 143 HLA-DRB1 and 47 HLA-DQB1 alleles were observed, which accounted for 6.98%, 7.06%, 6.46%, 9.11% and 7.91%, respectively, of the alleles in each locus in the world (IMGT 3.16 Release, Apr. 2014). Among the 100 most common haplotypes from the 169,995 individuals, nine distinct haplotypes displayed significant regionally specific distributions. Among these, three were predominant in the South China region (i.e., the 20th, 31st, and 81sthaplotypes), another three were predominant in the Southwest China region (i.e., the 68th, 79th, and 95th haplotypes), one was predominant in the South and Southwest China regions (the 18th haplotype), one was relatively common in the Northeast and North China regions (the 94th haplotype), and one was common in the Northeast, North and Northwest China (the 40th haplotype). In conclusion, this is the first to analyze high-resolution HLA diversities across the entire country of China, based on a detailed and complete data set that covered 31 provinces, autonomous regions, and municipalities. Specifically, we also evaluated the HLA matching probabilities within and between geographic regions and analyzed the regional differences in the HLA diversities in China. We believe that the data presented in this study might be useful for unrelated HLA-matched donor searches, donor registry planning, population genetic studies, and anthropogenesis

  20. Biochemical and functional analyses of gp130 mutants unveil JAK1 as a novel therapeutic target in human inflammatory hepatocellular adenoma

    PubMed Central

    Poussin, Karine; Pilati, Camilla; Couchy, Gabrielle; Calderaro, Julien; Bioulac-Sage, Paulette; Bacq, Yannick; Paradis, Valérie; Leteurtre, Emmanuelle; Sturm, Nathalie; Ramos, Jeanne; Guettier, Catherine; Bardier-Dupas, Armelle; Boulai, Anais; Wendum, Dominique; Selves, Janick; Izard, Tina; Nault, Jean-Charles; Zucman-Rossi, Jessica

    2013-01-01

    Inflammatory hepatocellular adenomas (IHCAs) are benign liver lesions that can be characterized histologically by the presence of an inflammatory infiltrate and at the molecular level by the overexpression of acute phase inflammatory response genes. Recurrent somatic mutations of the interleukin-6 (IL-6) signal transducer (IL6ST) locus, encoding the critical component of the IL-6 signal transduction machinery gp130, are present in 60% of IHCAs and in a subset (2%) of hepatocellular carcinoma (HCCs). By screening of 256 human hepatic adenoma specimens (the largest genetic analysis of IL6ST performed to date in this setting), we identified 24 distinct somatic IL6ST mutations among 66 mutant adenomas. The functional analysis of nine different gp130 mutants expressed in hepatic cancer cell lines consistently revealed the constitutive and IL-6-independent activation of the JAK/STAT signaling pathway. We further demonstrated that the signaling activity of mutant gp130 in IHCA remains responsive to suppressor of cytokine signaling 3 (SOCS3), a physiological gp130 inhibitor. Specifically, cells expressing a double mutant variant of gp130 with a disrupted SOCS3-binding site at residue 759 (Y186/Y759F) displayed a hyperactivation of signal transducer and activator of transcription 3 (STAT3) as compared with cells expressing the endogenous IHCA-associated Y186 gp130 mutant. Notably, we identified that constitutive signaling via gp130 in IHCA requires the Janus kinase family member JAK1, but not JAK2 or tyrosine kinase 2. In support of this notion, AG490, a tyrosine kinase inhibitor that selectively blocks JAK2, had no effect on gp130 activity. In stark contrast, we showed that ruxolitinib, a JAK1/JAK2-selective tyrosine kinase inhibitor used to treat patients with myelofibrosis, dramatically impaired JAK1-STAT signaling downstream of all IHCA-associated gp130 mutants. In conclusion, our findings provide a rationale for the use of JAK1 inhibitors for the treatment of HCAs

  1. Simulations and projections of major air pollutants over the United States and uncertainty analyses, effects of natural change and human activities

    NASA Astrophysics Data System (ADS)

    Lei, Hang

    Changes in global climate and pollutant emissions are very likely to continue in the coming decades driven by the human-related activities and natural fluctuations in the Earth climate system. These potential changes would have very important consequences on regional air quality over the contiguous United States due to their effects on atmospheric chemical and physical processes. To understand these effects, the present studies use the global climate chemistry model, CAM-Chem version 3, to systematically assess potential changes in major air pollutants including surface ozone, particulate matter and mercury from the present (1998--2002) to the 2050 (2048--2052). The projections of future air quality consider changes in global climate, precursor emissions from anthropogenic and biogenic sources, and pollutant transport. Moreover, to evaluate the projection uncertainties resulting from different plausible trends of climate and emissions as a result of unknown human-related activities and climate variations, three IPCC SRES scenarios, A1FI, A1B and B1, are considered and compared to evaluate the resulting uncertainty in projecting future pollutant concentrations. To achieve a better understanding on the effect of mineral dust emissions on changes in future air quality especially the PM concentrations, a physical dust aerosol module is developed and incorporated into the CAM-Chem model. A mercury module is developed for the CAM-Chem model to simulate the atmospheric cycle of mercury and its consequences on the toxicity of U.S. air quality. For the study of ozone air quality, we focus on the risk of high ozone episodes and the relative contributions from changes in local anthropogenic emissions (LE) versus changes in intercontinental transport (ICT) on 2050 U.S. surface ozone air quality. It is found that the projected changes in air temperature, precipitation, lighting, planetary boundary layer height and cyclone activities tend to intensify the associated extreme

  2. Anti-CD antibody microarray for human leukocyte morphology examination allows analyzing rare cell populations and suggesting preliminary diagnosis in leukemia.

    PubMed

    Khvastunova, Alina N; Kuznetsova, Sofya A; Al-Radi, Liubov S; Vylegzhanina, Alexandra V; Zakirova, Anna O; Fedyanina, Olga S; Filatov, Alexander V; Vorobjev, Ivan A; Ataullakhanov, Fazly

    2015-07-27

    We describe a method for leukocyte sorting by a microarray of anti-cluster-of-differentiation (anti-CD) antibodies and for preparation of the bound cells for morphological or cytochemical examination. The procedure results in a "sorted" smear with cells positive for certain surface antigens localised in predefined areas. The morphology and cytochemistry of the microarray-captured normal and neoplastic peripheral blood mononuclear cells are identical to the same characteristics in a smear. The microarray permits to determine the proportions of cells positive for the CD antigens on the microarray panel with high correlation with flow cytometry. Using the anti-CD microarray we show that normal granular lymphocytes and lymphocytes with radial segmentation of the nuclei are positive for CD3, CD8, CD16 or CD56 but not for CD4 or CD19. We also show that the described technique permits to obtain a pure leukemic cell population or to separate two leukemic cell populations on different antibody spots and to study their morphology or cytochemistry directly on the microarray. In cases of leukemias/lymphomas when circulating neoplastic cells are morphologically distinct, preliminary diagnosis can be suggested from full analysis of cell morphology, cytochemistry and their binding pattern on the microarray.

  3. A fast multiparameter MRI approach for acute stroke assessment on a 3T clinical scanner: preliminary results in a non-human primate model with transient ischemic occlusion

    PubMed Central

    Tong, Frank; Li, Chun-Xia; Yan, Yumei; Nair, Govind; Nagaoka, Tsukasa; Tanaka, Yoji; Zola, Stuart; Howell, Leonard

    2014-01-01

    Many MRI parameters have been explored and demonstrated the capability or potential to evaluate acute stroke injury, providing anatomical, microstructural, functional, or neurochemical information for diagnostic purposes and therapeutic development. However, the application of multiparameter MRI approach is hindered in clinic due to the very limited time window after stroke insult. Parallel imaging technique can accelerate MRI data acquisition dramatically and has been incorporated in modern clinical scanners and increasingly applied for various diagnostic purposes. In the present study, a fast multiparameter MRI approach including structural T1-weighted imaging (T1W), T2-weighted imaging (T2W), diffusion tensor imaging (DTI), T2-mapping, proton magnetic resonance spectroscopy, cerebral blood flow (CBF), and magnetization transfer (MT) imaging, was implemented and optimized for assessing acute stroke injury on a 3T clinical scanner. A macaque model of transient ischemic stroke induced by a minimal interventional approach was utilized for evaluating the multiparameter MRI approach. The preliminary results indicate the surgical procedure successfully induced ischemic occlusion in the cortex and/or subcortex in adult macaque monkeys (n=4). Application of parallel imaging technique substantially reduced the scanning duration of most MRI data acquisitions, allowing for fast and repeated evaluation of acute stroke injury. Hence, the use of the multiparameter MRI approach with up to five quantitative measures can provide significant advantages in preclinical or clinical studies of stroke disease. PMID:24834423

  4. Human Factors Engineering Data Base Development for Armored Combat Vehicles and Analyses of Three NATO Tank Systems. Volume 3. Human Factors Engineering Analysis of the British Chieftain Main Battle Tank.

    DTIC Science & Technology

    1986-07-03

    Commander’s Hatch nure -’. ommanoer- ; ar.cn. Haiter Handle. ind Comnat .-cc:, - .. .. m - . . . . ° : - THE BDM CORPORATION BDM/ABQ-86-0608-TR U 23 inches...3, 1986 8 0MIABQ-86-0608-TR I 86 8 7 089 Smm • ’ THE BDM CORPORATION 1801 Randoiph Road, S.E, Albuquerque, NM 87106 0 (505) 848-5000 _ _. I. * HUMAN...5 % 5-| THE BDM CORPORATION BDM/ABQ-86-O608-TR TABLE OF CONTENTSR Chapter Page I HUMAN FACTORS ENGINEERING ANALYSIS OF THE BRITISH CHIEFTAIN MAIN

  5. A preliminary study of the effects of individual response to challenge tests and stress induced by humans on learning performance of weaned piglets (Sus scrofa).

    PubMed

    Brajon, Sophie; Laforest, Jean-Paul; Schmitt, Océane; Devillers, Nicolas

    2016-08-01

    This study investigated whether individual behavioural characteristics of piglets and stress induced by experience with humans can influence learning performance. After weaning, piglets received a chronic experience with humans to modulate their emotional state: rough (ROU), gentle (GEN), or minimal (MIN) experience. Simultaneously, they were trained on a discrimination task. Afterward, their behaviour during challenge tests was assessed. The first learning step of the task involved associating a positive sound cue with a response (approach a trough) and success of piglets depended mostly on motivation to seek for reward. Although the experience with humans did not have direct effect, the degree of fear of handler, measured based on their reactivity to a human approach test, was related to motivation to seek rewards and learning speed of this first step in stressed ROU piglets, but not in MIN and GEN piglets. In contrast, the second learning step was more cognitively challenging, since it involved discrimination learning, including negative cues during which piglets had to learn to avoid the trough. Locomotion activity, measured during an open-field test, was associated with performance of the discrimination learning. To conclude, fearfulness towards humans and locomotion activity are linked with learning performance in relation to task complexity, highlighting the necessity to take into account these factors in animal research and management.

  6. High-potentiality preliminary selection criteria and transformation time-dependent factors analysis for establishing Epstein-Barr virus transformed human lymphoblastoid cell lines.

    PubMed

    Chang, I-C; Wu, J-Y; Lu, H-I; Ko, H-W; Kuo, J-L; Wang, C-Y; Shen, P-S; Hwang, S-M

    2006-12-01

    Infection of freshly isolated and cryopreserved lymphocytes with Epstein-Barr virus (EBV) leads to the establishment of human B lymphoblastoid cell lines (LCLs). Techniques for optimal infection of the lymphocytes are vital for the establishment of a human biobank. The present study found that more than half (58-86%) of such established LCLs had transport times of less than 48 h, cell densities exceeding 10(6) cells/ml and cell viabilities greater than 90%. After EBV infection, 3306 freshly isolated lymphocytes required 30.0 +/- 0.1 days to become LCLs. Conversely, 1210 cryopreserved lymphocytes required 36.2 +/- 0.4 days. Cell density and viability of the culture affected transformation time in freshly isolated lymphocytes. On the other hand, blood transport time, cryopreservation time and initial cell viability were major factors in cryopreserved specimens. These results contribute to general information concerning the establishment of a human biobank for EBV infected cells.

  7. Preliminary Investigation of Visual Attention to Human Figures in Photographs: Potential Considerations for the Design of Aided AAC Visual Scene Displays

    ERIC Educational Resources Information Center

    Wilkinson, Krista M.; Light, Janice

    2011-01-01

    Purpose: Many individuals with complex communication needs may benefit from visual aided augmentative and alternative communication systems. In visual scene displays (VSDs), language concepts are embedded into a photograph of a naturalistic event. Humans play a central role in communication development and might be important elements in VSDs.…

  8. Preliminary exploration of HLA-A 1101-restricted human cytomegalovirus glycoprotein B-specific CD8⁺ T cells in allogeneic stem-cell transplant recipients.

    PubMed

    Liu, Anbing; Hu, Jianhua; Wu, Wei; Huang, Yaping; Liang, Hanying; Wang, Huiqi; Yang, Rong; Fan, Jun

    2014-08-08

    T-cell responses directed against human cytomegalovirus (HCMV) glycoprotein B (gB) contribute to protective immunity against HCMV infection in both animal models and humans. However, the gB-specific human CD8(+) T cell responses remain poorly understood. gB antigen-specific CD8(+) T cells were stained with seven major histocompatibility complex (MHC)-peptide pentamers in 16 human leukocyte antigen (HLA)-A 1101-positive, HCMV-seropositive patients following hematopoietic stem cell transplantation (HSCT). Of these seven pentamers, the most frequent CD8(+) T-cell responses were directed against the gB332-340 peptide. These gB332-340-specific CD8(+) T cells were strongly associated with the presence of plasma HCMV immunoglobulin M in all HSCT recipients and exhibited a probable causal relationship with the level of pp65 antigenemia. Together, these data suggest a role for gB332-340-specific CD8(+) T cells in HCMV reactivation after HSCT. Furthermore, the pentamer assay may be valuable in detecting antigen-specific CD8(+) T cells.

  9. Building up analgesia in humans via the endogenous μ-opioid system by combining placebo and active tDCS: a preliminary report.

    PubMed

    DosSantos, Marcos F; Martikainen, Ilkka K; Nascimento, Thiago D; Love, Tiffany M; DeBoer, Misty D; Schambra, Heidi M; Bikson, Marom; Zubieta, Jon-Kar; DaSilva, Alexandre F

    2014-01-01

    Transcranial Direct Current Stimulation (tDCS) is a method of non-invasive brain stimulation that has been frequently used in experimental and clinical pain studies. However, the molecular mechanisms underlying tDCS-mediated pain control, and most important its placebo component, are not completely established. In this pilot study, we investigated in vivo the involvement of the endogenous μ-opioid system in the global tDCS-analgesia experience. Nine healthy volunteers went through positron emission tomography (PET) scans with [11C]carfentanil, a selective μ-opioid receptor (MOR) radiotracer, to measure the central MOR activity during tDCS in vivo (non-displaceable binding potential, BPND)--one of the main analgesic mechanisms in the brain. Placebo and real anodal primary motor cortex (M1/2mA) tDCS were delivered sequentially for 20 minutes each during the PET scan. The initial placebo tDCS phase induced a decrease in MOR BPND in the periaqueductal gray matter (PAG), precuneus, and thalamus, indicating activation of endogenous μ-opioid neurotransmission, even before the active tDCS. The subsequent real tDCS also induced MOR activation in the PAG and precuneus, which were positively correlated to the changes observed with placebo tDCS. Nonetheless, real tDCS had an additional MOR activation in the left prefrontal cortex. Although significant changes in the MOR BPND occurred with both placebo and real tDCS, significant analgesic effects, measured by improvements in the heat and cold pain thresholds, were only observed after real tDCS, not the placebo tDCS. This study gives preliminary evidence that the analgesic effects reported with M1-tDCS, can be in part related to the recruitment of the same endogenous MOR mechanisms induced by placebo, and that such effects can be purposely optimized by real tDCS.

  10. Crystallization and preliminary X-ray analysis of Na-ASP-1, a multi-domain pathogenesis-related-1 protein from the human hookworm parasite Necator americanus

    SciTech Connect

    Asojo, Oluwatoyin A.; Loukas, Alex; Inan, Mehmet; Barent, Rick; Huang, Jicai; Plantz, Brad; Swanson, Amber; Gouthro, Mark; Meagher, Michael M.; Hotez, Peter J.

    2005-04-01

    In order to clarify the structural basis of the pathogenesis-related-1 domain, Na-ASP-1, the first multi-domain ASP from the human hookworm parasite N. americanus, has been crystallized. 2.2 Å resolution data have been collected from a crystal belonging to the monoclinic space group P2{sub 1}. Human hookworm infection is a major cause of anemia and malnutrition in the developing world. In an effort to control hookworm infection, the Human Hookworm Vaccine Initiative has identified candidate vaccine antigens from the infective larval stage (L3) of the parasite, including a family of pathogenesis-related-1 (PR-1) proteins known as the ancylostoma-secreted proteins (ASPs). The functions of the ASPs are unknown. In addition, it is unclear why some ASPs have one while others have multiple PR-1 domains. There are no known structures of a multi-domain ASP and in an effort to remedy this situation, recombinant Na-ASP-1 has been expressed, purified and crystallized. Na-ASP-1 is a 406-amino-acid multi-domain ASP from the prevalent human hookworm parasite Necator americanus. Useful X-ray data to 2.2 Å have been collected from a crystal that belongs to the monoclinic space group P2{sub 1} with unit-cell parameters a = 67.7, b = 74.27, c = 84.60 Å, β = 112.12°. An initial molecular-replacement solution has been obtained with one monomer in the asymmetric unit.

  11. Effects of subinhibitory amounts of ampicillin, amoxycillin and mecillinam on the adhesion of Escherichia coli bacteria to human urinary tract epithelial cells: a preliminary study.

    PubMed

    Svanborg-Edén, C; Sandberg, T; Stenqvist, K; Ahlstedt, S

    1979-01-01

    Attachment to mucous surfaces may be a prerequisite for bacteria colonizing these surfaces or invading underlying tissues. Subinhibitory amounts of ampicillin and amoxycillin but not mecillinam decreased the attachment of Escherichia coli bacteria to human uro-epithelial cells in vitro. No significant synergistic effect on the attachment by the antibiotics was obtained. The present report indicates a new parameter for the study of antibacterial actions of drugs.

  12. Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the VP8* carbohydrate-binding protein of the human rotavirus strain Wa

    SciTech Connect

    Kraschnefski, Mark J.; Scott, Stacy A.; Holloway, Gavan; Coulson, Barbara S.; Itzstein, Mark von; Blanchard, Helen

    2005-11-01

    The carbohydrate-binding component (VP8*{sub 64–223}) of the human Wa rotavirus spike protein has been overexpressed in E. coli, purified and crystallized in two different crystal forms. X-ray diffraction data have been collected that have enabled determination of the Wa VP8*{sub 64–223} structure by molecular replacement. Rotaviruses exhibit host-specificity and the first crystallographic information on a rotavirus strain that infects humans is reported here. Recognition and attachment to host cells, leading to invasion and infection, is critically linked to the function of the outer capsid spike protein of the rotavirus particle. In some strains the VP8* component of the spike protein is implicated in recognition and binding of sialic-acid-containing cell-surface carbohydrates, thereby enabling infection by the virus. The cloning, expression, purification, crystallization and initial X-ray diffraction analysis of the VP8* core from human Wa rotavirus is reported. Two crystal forms (trigonal P3{sub 2}21 and monoclinic P2{sub 1}) have been obtained and X-ray diffraction data have been collected, enabling determination of the VP8*{sub 64–223} structure by molecular replacement.

  13. Bone natural autofluorescence and confocal laser scanning microscopy: Preliminary results of a novel useful tool to distinguish between forensic and ancient human skeletal remains.

    PubMed

    Capasso, Luigi; D'Anastasio, Ruggero; Guarnieri, Simone; Viciano, Joan; Mariggiò, Maria

    2017-03-01

    The fast, high-throughput distinction between palaeoanthropological/archaeological remains and recent forensic/clinical bone samples is of vital importance in the field of medico-legal science. In this paper, a novel dating method was developed using the autofluorescence of human bones and the confocal laser scanning microscope as the means to distinguish between archaeological and forensic anthropological skeletal findings. Human bones exhibit fluorescence, typically induced by natural antibiotics that are absorbed by collagen, and provide secondary, exogenous fluorophores. However, primary natural fluorescence (or autofluorescence) caused by enigmatic endogenous fluorophores is also present as a micro-phenomenon, whose nature is still obscure. Here, we show that the endogenous fluorophores are mucopolysaccharides of the Rouget-Neumann sheath and, more relevant, that the intensity of the natural fluorescence in human bone decreases in a relationship to the antiquity of the samples. These results suggest that the autofluorescence of bone is a promising technique for the assessment of skeletal remains that may be potentially of medico-legal interest. A larger study is proposed to confirm these findings and to create a predictive model between the autofluorescence intensity and the time since death.

  14. Characteristics and Preliminary Observations of the Influence of Electromyostimulation on the Size and Function of Human Skeletal Muscle During 30 Days of Simulated Microgravity

    NASA Technical Reports Server (NTRS)

    Duvoisin, Marc R.; Convertino, Victor A; Buchanan, Paul; Gollinick, Philip D.; Dudley, Gary A.

    1989-01-01

    During 30 days (d) of bedrest, the practicality of using Elec- troMyoStimulation (EMS) as a deterrent to atrophy and strength loss of lower limb musculature was examined. An EMS system was developed that provided variable but quantifiable levels of EMS, and measured torque. The dominant log of three male subjects was stimulated twice daily in a 3-d on/1-d off cycle during bedrest. The non-dominant leg of each subject acted as a control. A stimulator, using a 0.3 ms monophasic 60 Hz pulse waveform, activated muscle tissue for 4 s. The output waveform from the stimulator was sequenced to the Knee Extensors (KE), Knee Flex- ors (KF), Ankle Extensors (AE), and Ankle Flexors (AF), and caused three isometric contractions of each muscle group per minute. Subject tolerance determined EMS Intensity. Each muscle group received four 5-min bouts of EMS each session with a 10 -min rest between bouts. EMS and torque levels for each muscle action were recorded directly an a computer. Overall average EMS Intensity was 197, 197, 195, and 188 mA for the KE, KF, AF, and AE, respectively. Overall average torque development for these muscle groups was 70, 16, 12, and 27 Nm, respectively. EMS intensity doubled during the study, and average torque increased 2.5 times. Average maximum torque throughout a session reached 54% of maximal voluntary for the KE and 29% for the KF. Reductions in leg volume, muscle compartment size, cross-sectional area of slow and fast-twitch fibers, strength, and aerobic enzyme activities, and increased log compliance were attenuated in the legs which received EMS during bedrest. These results indicate that similar EMS levels induce different torques among different muscle groups and that repeated exposure to EMS increases tolerance and torque development. Longer orien- tation periods, therefore, may enhance its effectiveness. Our preliminary data suggest that the efficacy of EMS as an effective countermeasure for muscle atrophy and strength loss during long

  15. Intracranial hemorrhage alters scalp potential distribution in bioimpedance cerebral monitoring: Preliminary results from FEM simulation on a realistic head model and human subjects

    PubMed Central

    Atefi, Seyed Reza; Seoane, Fernando; Kamalian, Shervin; Rosenthal, Eric S.; Lev, Michael H.; Bonmassar, Giorgio

    2016-01-01

    Purpose: Current diagnostic neuroimaging for detection of intracranial hemorrhage (ICH) is limited to fixed scanners requiring patient transport and extensive infrastructure support. ICH diagnosis would therefore benefit from a portable diagnostic technology, such as electrical bioimpedance (EBI). Through simulations and patient observation, the authors assessed the influence of unilateral ICH hematomas on quasisymmetric scalp potential distributions in order to establish the feasibility of EBI technology as a potential tool for early diagnosis. Methods: Finite element method (FEM) simulations and experimental left–right hemispheric scalp potential differences of healthy and damaged brains were compared with respect to the asymmetry caused by ICH lesions on quasisymmetric scalp potential distributions. In numerical simulations, this asymmetry was measured at 25 kHz and visualized on the scalp as the normalized potential difference between the healthy and ICH damaged models. Proof-of-concept simulations were extended in a pilot study of experimental scalp potential measurements recorded between 0 and 50 kHz with the authors’ custom-made bioimpedance spectrometer. Mean left–right scalp potential differences recorded from the frontal, central, and parietal brain regions of ten healthy control and six patients suffering from acute/subacute ICH were compared. The observed differences were measured at the 5% level of significance using the two-sample Welch t-test. Results: The 3D-anatomically accurate FEM simulations showed that the normalized scalp potential difference between the damaged and healthy brain models is zero everywhere on the head surface, except in the vicinity of the lesion, where it can vary up to 5%. The authors’ preliminary experimental results also confirmed that the left–right scalp potential difference in patients with ICH (e.g., 64 mV) is significantly larger than in healthy subjects (e.g., 20.8 mV; P < 0.05). Conclusions: Realistic

  16. Production of a monoclonal antibody to a membrane antigen of human T-cell leukaemia virus (HTLV1/ATLV)-infected cell lines from a systemic lupus erythematosus (SLE) patient: serological analyses for HTLV1 infections in SLE patients.

    PubMed Central

    Kurata, A; Katamine, S; Fukuda, T; Mine, M; Ikari, N; Kanazawa, H; Matsunaga, M; Eguchi, K; Nagataki, S

    1985-01-01

    Human T-cell leukaemia virus (HTLV1/ATLV), which causes adult T cell leukaemia (ATL), is an infectious, lymphotrophic retrovirus unique for humans. The present study was undertaken to determine whether HTLV1 had any pathogenetic role for systemic lupus erythematosus (SLE). The incidence of antibodies to ATL cell-associated antigens (ATLA) in sera from patients with SLE and other collagen diseases was investigated by an indirect immunofluorescent cytoplasmic staining of an HTLV1-infected cell line (MT-1). A radioimmunoassay was also performed to detect antibodies to HTLV1 protein and crude membrane fraction derived from an HTLV1-producing cell line MT-2. Furthermore, an Epstein-Barr virus (EBV)-transformed B cell line (ES-1) was constructed from an SLE patient, which produced a monoclonal antibody (IgG, lambda) reactive to an HTLV1-related cell-membrane antigen expressed on MT-1 and MT-2 cells. The specific reactivity of the monoclonal antibody was analysed by an indirect immunofluorescent cell-membrane staining and a microcytotoxicity test. The incidence of anti-ATLA antibodies was not different among SLE and other collagen diseases. The monoclonal antibody produced by ES-1 stained and killed HTLV1-infected cell lines specifically, but did not react with other human lymphoid cell lines. This monoclonal antibody failed to react with peripheral blood mononuclear cells (PBMC), mitogen-induced T cell blasts, and iododeoxyuridine-treated T cells from SLE patients. Thus, a possible role of HTLV1 in the aetiology of SLE was not established. PMID:2998659

  17. Human genomic variation

    PubMed Central

    Disotell, Todd R

    2000-01-01

    The recent completion and assembly of the first draft of the human genome, which combines samples from several ethnically diverse males and females, provides preliminary data on the extent of human genetic variation. PMID:11178257

  18. Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human histidine triad nucleotide-binding protein 2 (hHINT2)

    PubMed Central

    Dolot, Rafał; Włodarczyk, Artur; Bujacz, Grzegorz D.; Nawrot, Barbara

    2013-01-01

    Histidine triad nucleotide-binding protein 2 (HINT2) is a mitochondrial adenosine phosphoramidase mainly expressed in the pancreas, liver and adrenal gland. HINT2 possibly plays a role in apoptosis, as well as being involved in steroid biosynthesis, hepatic lipid metabolism and regulation of hepatic mitochondria function. The expression level of HINT2 is significantly down-regulated in hepatocellular carcinoma patients. To date, endogenous substrates for this enzyme, as well as the three-dimensional structure of human HINT2, are unknown. In this study, human HINT2 was cloned, overexpressed in Escherichia coli and purified. Crystallization was performed at 278 K using PEG 4000 as the main precipitant; the crystals, which belonged to the tetragonal space group P41212 with unit-cell parameters a = b = 76.38, c = 133.25 Å, diffracted to 2.83 Å resolution. Assuming two molecules in the asymmetric unit, the Matthews coefficient and the solvent content were calculated to be 2.63 Å3 Da−1 and 53.27%, respectively. PMID:23832208

  19. Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human histidine triad nucleotide-binding protein 2 (hHINT2).

    PubMed

    Dolot, Rafał; Włodarczyk, Artur; Bujacz, Grzegorz D; Nawrot, Barbara

    2013-07-01

    Histidine triad nucleotide-binding protein 2 (HINT2) is a mitochondrial adenosine phosphoramidase mainly expressed in the pancreas, liver and adrenal gland. HINT2 possibly plays a role in apoptosis, as well as being involved in steroid biosynthesis, hepatic lipid metabolism and regulation of hepatic mitochondria function. The expression level of HINT2 is significantly down-regulated in hepatocellular carcinoma patients. To date, endogenous substrates for this enzyme, as well as the three-dimensional structure of human HINT2, are unknown. In this study, human HINT2 was cloned, overexpressed in Escherichia coli and purified. Crystallization was performed at 278 K using PEG 4000 as the main precipitant; the crystals, which belonged to the tetragonal space group P41212 with unit-cell parameters a = b = 76.38, c = 133.25 Å, diffracted to 2.83 Å resolution. Assuming two molecules in the asymmetric unit, the Matthews coefficient and the solvent content were calculated to be 2.63 Å(3) Da(-1) and 53.27%, respectively.

  20. Purification, crystallization and preliminary structural characterization of the N-terminal region of the human formin-homology protein FHOD1

    SciTech Connect

    Schulte, Antje Rak, Alexey; Pylypenko, Olena; Ludwig, Diana; Geyer, Matthias

    2007-10-01

    The N-terminal region (1–339) of the human FHOD1 protein has been crystallized in two different crystal forms. A crystal of the (C31S,C71S) mutant diffracted to around 2.3 Å resolution. Formins are key regulators of actin cytoskeletal dynamics that constitute a diverse protein family that is present in all eukaryotes examined. They typically consist of more than 1000 amino acids and are defined by the presence of two conserved regions, namely the formin homology 1 and 2 domains. Additional conserved domains comprise a GTPase-binding domain for activation, a C-terminal autoregulation motif and an N-terminal recognition domain. In this study, the N-terminal region (residues 1–339) of the human formin homology domain-containing protein 1 (FHOD1) was purified and crystallized from 20%(w/v) PEG 4000, 10%(v/v) glycerol, 0.3 M magnesium chloride and 0.1 M Tris–HCl pH 8.0. Native crystals belong to space group P1, with unit-cell parameters a = 35.4, b = 73.9, c = 78.7 Å, α = 78.2, β = 86.2, γ = 89.7°. They contain two monomers of FHOD1 in the asymmetric unit and diffract to a resolution of 2.3 Å using a synchrotron-radiation source.

  1. Reduction in deposition of indium 111-labeled platelets after autologous endothelial cell seeding of Dacron aortic bifurcation grafts in humans: a preliminary report

    SciTech Connect

    Ortenwall, P.; Wadenvik, H.; Kutti, J.; Risberg, B.

    1987-07-01

    Autologous endothelial seeding (AES) of vascular prostheses in dogs increases thrombus-free surface and improves prosthetic prostacyclin production, patency, and the ability to withstand hematogenous challenge with bacteria. No such information is available in human subjects. In the present study one limb of an aortic Dacron bifurcation prosthesis was seeded with autologous endothelial cells (ECs) harvested from the distal portion of the saphenous vein by enzymatic treatment. The deposition of indium 111-labeled platelets on the vascular prostheses was studied 1 and 4 months after operation. In seven of nine patients seeding resulted in decreased accumulation of radiolabeled platelets compared with sham-seeded control limbs (p less than 0.04), when studied 1 month after surgery. A decrease in platelet accumulation occurred over the whole prosthesis between 1 and 4 months, and no significant difference was noted at 4 months between seeded and nonseeded graft limbs. Although the seeding density was very low (440 ECs/cm2), the observed difference in platelet accumulation for AES-treated graft limbs in the early postoperative course merits further investigation of this technique in human beings.

  2. Crystallization and preliminary X-ray diffraction of human interleukin-7 bound to unglycosylated and glycosylated forms of its α-receptor

    SciTech Connect

    Wickham, Joseph Jr; Walsh, Scott T. R.

    2007-10-01

    Bacterial and insect cell expression systems have been developed to produce unglycosylated and glycosylated forms of human interleukin-7 (IL-7) and the extracellular domain of its α receptor, IL-7Rα. We report the crystallization and X-ray diffraction of IL-7 complexes to both unglycosylated and glycosylated forms of the IL-7Rα to 2.7 and 3.0 Å, respectively. The interleukin-7 (IL-7) signaling pathway plays an essential role in the development, proliferation and homeostasis of T and B cells in cell-mediated immunity. Understimulation and overstimulation of the IL-7 signaling pathway leads to severe combined immunodeficiency, autoimmune reactions, heart disease and cancers. Stimulation of the IL-7 pathway begins with IL-7 binding to its α-receptor, IL-7Rα. Protein crystals of unglycosylated and glycosylated complexes of human IL-7–IL-7Rα extracellular domain (ECD) obtained using a surface entropy-reduction approach diffract to 2.7 and 3.0 Å, respectively. Anomalous dispersion methods will be used to solve the unglycosylated IL-7–IL-7Rα ECD complex structure and this unglycosylated structure will then serve as a model in molecular-replacement attempts to solve the structure of the glycosylated IL-7–α-receptor complex.

  3. Characterization and Cytotoxicity Analysis of a Ciprofloxacin Loaded Chitosan/Bioglass Scaffold on Cultured Human Periodontal Ligament Stem Cells: a Preliminary Report

    PubMed Central

    Abdelfattah, Maha I.; Nasry, Sherine A.; Mostafa, Amani A.

    2016-01-01

    AIM: The aim of this study was to analyze the cytotoxicity of ciprofloxacin (CIP) loaded on chitosan bioactive glass scaffold on human periodontal ligament stem cells (PLSCs) in vitro. MATERIALS AND METHODS: PLSCs obtained from human third molars, cultures treated with medium containing 15 x 15 mm chitosan/bioactive glass scaffolds without/with different concentration 0, 5, 10, and 20 % of CIP. A total of 15 x 10^3 cells were plated in 6 well plates. The attached cells of each group were harvested from the plates after 1, 4 and 8 days of culture to detect the viability of cells. The cell number was determined using a hemocytometer and the trypan blue dye-exclusion assay. Data was analyzed using normality using Shapiro-Wilk test. Comparisons between groups were made using One-way ANOVA complemented by Tukey’s test. RESULTS: When comparing the proliferation rate of cells in the four groups, no statistically significant difference was found (P = 0.633). With regards to cell viability, no statistical difference was found between the 0, 5, and 10 % CIP concentrations, while the 20 % CIP concentration demonstrated the least viability with a high statistically significant difference (P = 0.003). CONCLUSION: Twenty percentages CIP demonstrated the least proliferation rate and viability. PMID:27703576

  4. Preliminary Study on Gene Expression of Chitinase-Like Cytokines in Human Airway Epithelial Cell Under Chitin and Chitosan Microparticles Treatment.

    PubMed

    Alimohammadi, Masumeh; Yeganeh, Farshid; Haji Molla Hoseini, Mostafa

    2016-06-01

    Small-sized chitin and chitosan microparticles (MPs) reduce allergic inflammation. We examined the capacity of these glycans to stimulate A549 human airway epithelial cells to determine the feasibility of using of these glycans as allergic therapeutic modality. A549 cells were treated with MPs and then expressions levels of chitinase domain-containing 1 (CHID1) and chitinase 3-like 1 (CHI3L1) genes were determined by quantitative real-time PCR. IL-6 production was measured by ELISA. Chitin MPs resulted in upregulation of CHI3L1 expression by 35.7-fold while mRNA expression did not change with chitosan MPs. Compared to the untreated group, production of IL-6 was significantly decreased in the chitosan MPs-treated group, but chitin MPs treatment cause elevation of IL-6 level. This study demonstrates that chitin potently induces CHI3L1 expression, but chitosan is relatively inert. This effect and inhibition of pro-inflammatory cytokine (IL-6) suggest that chitosan MPs may possess more potential for therapeutic uses in human airway allergic inflammation.

  5. Preliminary investigation of human serum albumin-Vβ inhibition on toxic shock syndrome induced by staphylococcus enterotoxin B in vitro and in vivo.

    PubMed

    Yuan, Qifeng; Li, Lin; Pian, Yaya; Hao, Huaijie; Zheng, Yuling; Zang, Yating; Jiang, Hua; Jiang, Yongqiang

    2016-04-01

    Staphylococcus enterotoxin B (SEB) is a superantigen that can induce massive activation of T cells with specific Vβ and inflammatory cytokine cascades, which mediate shock. To date, no SEB vaccine has been developed for preventing toxic shock syndrome (TSS). Here, we evaluated the therapeutic effect of a fusion protein human serum albumin-Vβ (HSA-Vβ) on TSS induced by SEB. Compared with Vβ, the preparation of HSA-Vβ was much easier to handle owing to its solubility. Affinity testing showed that HSA-Vβ had high affinity for SEB. In vitro results showed that HSA-Vβ could effectively inhibit interferon (IFN)-γ and tumor necrosis factor (TNF)-α secretion by human peripheral blood mononuclear cells. Moreover, in vivo, HSA-Vβ reduced IFN-γ and TNF-α levels in the serum and protected mice from SEB lethal challenge when administered simultaneously with SEB or 30 min after SEB. In summary, we simplified the preparation of Vβ by fusion with HSA, creating the HSA-Vβ protein, which effectively inhibited cytokine production and protected mice from lethal challenge with SEB. These data indicated that HSA-Vβ may represent a novel therapeutic strategy for the treatment of SEB-induced TSS.

  6. Preliminary Results Concerning the Influence of Human Kisspeptin on LH Secretion in Prussian Carp (Carassius gibelio) Females at the Stage of Ovarian Recrudescence and Spawning Season.

    PubMed

    Gosiewski, Grzegorz; Sokolowska-Mikolajczyk, Miroslawa; Chyb, Jaroslaw; Socha, Magdalena

    2015-01-01

    The aim of the experiments was to evaluate the influence of human kisspeptin on LH secretion of Prussian carp (Carassius gibelio) females during ovarian recrudescence and spawning season. For the experiments, human kisspeptin KISS1 (0.1 mg kg(-1) of body weight--b.w.), GnRH analogue (Des Gly(10), D-Ala(6)) GnRH-A (20 µg kg(-1) b.w.) and dopamine antagonist (pimozide) (5 mg kg(-1) b.w.) were used alone or in combinations. At 3, 6, 12, 24 hours after injection(s) blood samples were collected from all fish. LH levels were measured in plasma with the use of the ELISA method. KISS1 did not show any significant effects on spontaneous LH secretion in both tested seasons. At 12 hours sampling time (both stages of gonad maturity) a combination of tested compounds (GnRH-A+KISS1) significantly increased LH release in comparison with the control. In the stage of gonad recrudescence KISS1 significantly increased LH secretion evoked by pimozide at 24 hours. A combination of three components: KISS1, GnRH-A, and pimozide significantly decreased LH secretion in comparison to LH secretion evoked by GnRH-A and pimozide during stage of gonad recrudescence. These results suggest that kisspeptin is involved in seasonal control of reproduction in Prussian carp. The possible interaction of kisspeptin and the dopaminergic system is also discussed.

  7. Preliminary Drill Sites

    DOE Data Explorer

    Lane, Michael

    2013-06-28

    Preliminary locations for intermediate depth temperature gradient holes and/or resource confirmation wells based on compilation of geological, geophysical and geochemical data prior to carrying out the DOE-funded reflection seismic survey.

  8. Characteristics and preliminary observations of the influence of electromyostimulation on the size and function of human skeletal muscle during 30 days of simulated microgravity

    NASA Technical Reports Server (NTRS)

    Duvoisin, Marc R.; Convertino, Victor A.; Buchanan, Paul; Gollnick, Philip A.; Dudley, Gary A.

    1989-01-01

    The effect of transcutaneous electromyostimulation (EMS) on the development of atrophy and the loss of strength in lower limb musculature in humans exposed to microgravity was determined in three subjects who received EMS twice daily in a 3-d on/1-d off cycle on their dominant leg during 30 days of bedrest. The output waveform from the stimulator was sequenced to the knee extensors, knee flexors, ankle extensors, and ankle flexors, and caused three isometric contractions of each muscle group per minute. It was found that, in the dominant leg, EMS acted to attenuate the changes caused by bedrest, such as reductions in the leg volume, muscle compartment size, cross-sectional area of slow- and fast-twitch fibers, strength, and aerobic enzyme activities, and an increase in leg compliance.

  9. Preliminary paleoecological reconstruction of long-term relationship between human and environment in the northern part of Danube-along Plain, Hungary

    NASA Astrophysics Data System (ADS)

    Kustár, Rozália; Molnár, Dávid; Sümegi, Pál; Törőcsik, Tünde; Sávai, Szilvia

    2016-06-01

    The peat bog at Ócsa is located at the northern part of the Danube-Tisa Interfluves at the transitional zone of two landscapes with different morphological characters. At the boundary of the Danube-Tisa Interfluves and the Danube-along Plain a marshland sequence can be found from Hajós to Ócsa. We extended our research to the Ócsa peat bog to complete the environmental historical investigations in the examined area, as well. The bog is located in a former pool formed by the Danube River in which aeolian sand and thick lake sediment deposited from the Late Pleistocene. The initial oligotrophic lake became mesotrophic, therefore thick carbonate sediment deposited. Afterwards, as a consequence of the Neolithic human occupations, the natural development of the lake changed drastically and the lake choked up. The pollen and quartermalacological analysis of the area support the mentioned geological processes.

  10. Expression, purification, crystallization and preliminary X-ray analysis of the human RuvB-like protein RuvBL1

    SciTech Connect

    Gorynia, Sabine; Gonçalves, Susana; Coelho, Ricardo; Lopes, Gonçalo; Thomaz, Mónica; Huber, Martina; Donner, Peter

    2006-01-01

    Human RuvB-like protein RuvBL1 plays important roles in essential signaling pathways like c-Myc and Wnt, in transcription, and in DNA repair and apoptosis. Crystals of both native and a Se-Met derivative were obtained and characterized. SAD data leading to the structure solution at 2.2 Å were measured from the Se-Met crystals. RuvBL1, an evolutionary highly conserved protein related to the AAA{sup +} family of ATPases, has been crystallized using the hanging-drop vapour-diffusion method at 293 K. The crystals are hexagonal and belong to space group P6, with unit-cell parameters a = b = 207.1, c = 60.7 Å and three molecules in the asymmetric unit.

  11. Novel hybrids of metronidazole and quinolones: synthesis, bioactive evaluation, cytotoxicity, preliminary antimicrobial mechanism and effect of metal ions on their transportation by human serum albumin.

    PubMed

    Cui, Sheng-Feng; Peng, Li-Ping; Zhang, Hui-Zhen; Rasheed, Syed; Vijaya Kumar, Kannekanti; Zhou, Cheng-He

    2014-10-30

    A novel series of hybrids of metronidazole and quinolones as antimicrobial agents were designed and synthesized. Most prepared compounds exhibited good or even stronger antimicrobial activities in comparison with reference drugs. Furthermore, these highly active metronidazole-quinolone hybrids showed appropriate ranges of pKa, log P and aqueous solubility to pharmacokinetic behaviors and no obvious toxicity to A549 and human hepatocyte LO2 cells. Their competitive interactions with metal ions to HSA revealed that the participation of Mg(2+) ion in compound 7d-HSA association could result in a concentration increase of free compound 7d. Molecular modeling and experimental investigation of compound 7d with DNA suggested that possible antibacterial mechanism might be in relation with multiple binding sites between bioactive molecules and topo IV-DNA complex.

  12. Small interfering RNA-mediated silencing of nicotinamide phosphoribosyltransferase (NAMPT) and lysosomal trafficking regulator (LYST) induce growth inhibition and apoptosis in human multiple myeloma cells: A preliminary study.

    PubMed

    Bong, Ivyna Pau Ni; Ng, Ching Ching; Fakiruddin, Shaik Kamal; Lim, Moon Nian; Zakaria, Zubaidah

    2016-11-10

    Multiple myeloma (MM) is a malignancy of B lymphocytes or plasma cells. Our array-based comparative genomic hybridization findings revealed chromosomal gains at 7q22.3 and 1q42.3, where nicotinamide (NAM) phosphoribosyltransferase (NAMPT) and lysosomal trafficking regulator (LYST) genes are localized, respectively. This led us to further study the functions of these genes in myeloma cells. NAMPT is a key enzyme involved in nicotinamide adenine dinucleotide salvage pathway, and it is frequently overexpressed in human cancers. In contrast, little is known about the function of LYST in cancer. The expression of LYST is shown to affect lysosomal size, granule size, and autophagy in human cells. In this study, the effects of small interfering RNA (siRNA)-mediated silencing of NAMPT and LYST on cell proliferation and apoptosis were evaluated in RPMI 8226 myeloma cells. Transfection efficiencies were determined by quantitative real time reverse transcriptase PCR. Cell proliferation was determined using MTT assay, while apoptosis was analyzed with flow cytometry using Annexin V-fluorescein isothiocyanate/propidium iodide assay. The NAMPT protein expression in siRNA-treated cells was estimated by enzyme-linked immunosorbent assay. Our results showed that NAMPT and LYST were successfully knockdown by siRNA transfection (p < 0.05). NAMPT or LYST gene silencing significantly inhibited cell proliferation and induced apoptosis in RPMI 8226 cells (p < 0.05). Silencing of NAMPT gene also decreased NAMPT protein levels (p < 0.01). Our study demonstrated that NAMPT and LYST play pivotal roles in the molecular pathogenesis of MM. This is the first report describing the possible functions of LYST in myelomagenesis and its potential role as a therapeutic target in MM.

  13. Small interfering RNA-mediated silencing of nicotinamide phosphoribosyltransferase (NAMPT) and lysosomal trafficking regulator (LYST) induce growth inhibition and apoptosis in human multiple myeloma cells: A preliminary study

    PubMed Central

    Bong, Ivyna Pau Ni; Ng, Ching Ching; Fakiruddin, Shaik Kamal; Lim, Moon Nian; Zakaria, Zubaidah

    2016-01-01

    Multiple myeloma (MM) is a malignancy of B lymphocytes or plasma cells. Our array-based comparative genomic hybridization findings revealed chromosomal gains at 7q22.3 and 1q42.3, where nicotinamide (NAM) phosphoribosyltransferase (NAMPT) and lysosomal trafficking regulator (LYST) genes are localized, respectively. This led us to further study the fprotein expression in unctions of these genes in myeloma cells. NAMPT is a key enzyme involved in nicotinamide adenine dinucleotide salvage pathway, and it is frequently overexpressed in human cancers. In contrast, little is known about the function of LYST in cancer. The expression of LYST is shown to affect lysosomal size, granule size, and autophagy in human cells. In this study, the effects of small interfering RNA (siRNA)-mediated silencing of NAMPT and LYST on cell proliferation and apoptosis were evaluated in RPMI 8226 myeloma cells. Transfection efficiencies were determined by quantitative real time reverse transcriptase PCR. Cell proliferation was determined using MTT assay, while apoptosis was analyzed with flow cytometry using Annexin V-fluorescein isothiocyanate/propidium iodide assay. The NAMPT protein expression in siRNA-treated cells was estimated by enzyme-linked immunosorbent assay. Our results showed that NAMPT and LYST were successfully knockdown by siRNA transfection (p < 0.05). NAMPT or LYST gene silencing significantly inhibited cell proliferation and induced apoptosis in RPMI 8226 cells (p < 0.05). Silencing of NAMPT gene also decreased NAMPT protein levels (p < 0.01). Our study demonstrated that NAMPT and LYST play pivotal roles in the molecular pathogenesis of MM. This is the first report describing the possible functions of LYST in myelomagenesis and its potential role as a therapeutic target in MM. PMID:27754828

  14. Molecular cytogenetic analyses of HIG, a novel human cell line carrying t(1;3)(p36.3;q25.3) established from a patient with chronic myelogenous leukemia in blastic crisis.

    PubMed

    Hosoya, Noriko; Ogawa, Seishi; Motokura, Tohru; Hangaishi, Akira; Wang, Lili; Qiao, Ying; Nannya, Yasuhito; Kogi, Mieko; Hirai, Hisamaru

    2003-12-01

    Chromosomal abnormalities involving 1p36, 3q21, and/or 3q26 have been reported in a subset of myeloid neoplasms having characteristic dysmegakaryopoiesis, and the overexpression of EVI1 on 3q26 or of MEL1 on 1p36 has been implicated in their pathogenesis. We describe molecular cytogenetic analyses of a novel human cell line, HIG, established from a unique case in which a novel translocation t(1;3)(p36;q26) appeared as the sole additional chromosomal abnormality at the time of blastic transformation of chronic myelogenous leukemia. The patient displayed clinical features resembling those of the 3q21q26 syndrome. The HIG cell line retained der(1)t(1;3)(p36;q26) but lost t(9;22)(q34;q11). To identify the relevant gene that would be deregulated by this translocation, we molecularly cloned the translocation's breakpoints. They were distant from the breakpoint cluster regions of the 3q21q26 syndrome or t(1;3)(p36;q21), and neither the EVI1 nor the MEL1 transcript was detected in the HIG cell line. None of the genes located within 150 kilobase pairs of the breakpoints were aberrantly expressed, suggesting that in this case other gene(s) more distant from the breakpoints are deregulated by possible remote effects. Further analyses of the deregulated genes in the HIG cell line should provide important insight into the mechanisms involved in these types of leukemias.

  15. Recombinant pollen allergens from Dactylis glomerata: preliminary evidence that human IgE cross-reactivity between Dac g II and Lol p I/II is increased following grass pollen immunotherapy.

    PubMed

    Roberts, A M; Van Ree, R; Cardy, S M; Bevan, L J; Walker, M R

    1992-07-01

    We previously described the isolation of three identical complementary DNA (cDNA) clones, constructed from Orchard/Cocksfoot grass (Dactylis glomerata) anther messenger RNA (mRNA), expressing a 140,000 MW beta-galactosidase fusion protein recognized by IgE antibodies in atopic sera. Partial nucleotide sequencing and inferred amino acid sequence showed greater than 90% homology with the group II allergen from Lolium perenne (Lol II) indicating they encode the group II equivalent, Dac g II. Western blot immunoprobing of recombinant lysates with rabbit polyclonal, mouse monoclonal and human polyclonal antisera demonstrates immunological identity between recombinant Dac g II, Lol p I and Lol p II. Similar cross-identity is observed with pollen extracts from three other grass species: Festuca rubra, Phleum pratense and Anthoxanthum odoratum. Recombinant Dac g II was recognized by species- and group-cross-reactive human IgE antibodies in 33% (4/12) of sera randomly selected from grass-sensitive individuals and in 67% (14/21) of sera from patients receiving grass pollen immunotherapy, whilst 0/4 sera from patients receiving venom immunotherapy alone contained Dac g II cross-reactive IgE. Cross-reactive IgG4 antibodies were detectable in 95% of sera from grass pollen immunotherapy patients. These preliminary data suggest that conventional grass pollen allergoid desensitization immunotherapy may induce IgE responses to a cross-reactive epitope(s) co-expressed by grass pollen groups I and II (and possibly group III) allergens.

  16. Identification and Preliminary Validation of Radiation Response Protein(s) in Human Blood for a High-throughput Molecular Biodosimetry Technology for the Future

    PubMed Central

    Nongrum, Saibadaiahun; Vaiphei, S. Thangminlal; Keppen, Joshua; Ksoo, Mandahakani; Kashyap, Ettrika; Sharan, Rajesh N.

    2017-01-01

    The absence of a rapid and high-throughput technology for radiation biodosimetry has been a great obstacle in our full preparedness to cope with large-scale radiological incidents. The existing cytogenetic technologies have limitations, primarily due to their time-consuming methodologies, which include a tissue culture step, and the time required for scoring. This has seriously undermined its application in a mass casualty scenario under radiological emergencies for timely triage and medical interventions. Recent advances in genomics and proteomics in the postgenomic era have opened up new platforms and avenues to discover molecular biomarkers for biodosimetry in the future. Using a genomic-to-proteomic approach, we have identified a basket of twenty “candidate” radiation response genes (RRGs) using DNA microarray and tools of bioinformatics immediately after ex vivo irradiation of freshly drawn whole blood of consenting and healthy human volunteers. The candidate RRGs have partially been validated using real-time quantitative polymerase chain reaction (RT-qPCR or qPCR) to identify potential “candidate” RRGs at mRNA level. Two potential RRGs, CDNK1A and ZNF440, have so far been identified as genes with potentials to form radiation response proteins in liquid biopsy of blood, which shall eventually form the basis of fluorescence- or ELISA-based quantitative immunoprobe assay for a high-throughput technology of molecular biodosimetry in the future. More work is continuing. PMID:28250912

  17. Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the N-­terminal carbohydrate-recognition domain of human galectin-4

    PubMed Central

    Zimbardi, Ana Lucia L. R.; Pinheiro, Matheus P.; Dias-Baruffi, Marcelo; Nonato, M. Cristina

    2010-01-01

    Galectin-4 is a tandem-repeat-type galectin that is expressed in the epithelium of the alimentary tract from the tongue to the large intestine. Additionally, strong expression of galectin-4 can also be induced in cancers in other tissues, including the breast and liver. In order to explore its potential as a target for anticancer drug design, elucidation of the structural basis of the carbohydrate-binding specificities of galectin-4 has been focused on. As an initial step, the N-­terminal carbohydrate-recognition domain of human galectin-4 (hGal4-CRD-­1) has been successfully crystallized using the vapour-diffusion technique, a complete data set has been collected to 2.2 Å resolution and the structure has been solved by the molecular-replacement technique. The crystals belonged to space group P6122, with unit-cell parameters a = b = 71.25, c = 108.66 Å. The asymmetric unit contained one molecule of hGal4-CRD-1, with a V M value of 2.34 Å3 Da−1 and a solvent content of 47.51%. PMID:20445255

  18. Crystallization and preliminary X-ray diffraction analysis of the complex between a human anti-alpha toxin antibody fragment and alpha toxin.

    PubMed

    Oganesyan, Vaheh; Barnes, Arnita; Tkaczyk, Christine; Ferguson, Andrew; Wu, Herren; Dall'Acqua, William F

    2013-03-01

    Staphylococcus aureus alpha toxin (AT) has been crystallized in complex with the Fab fragment of a human antibody (MEDI4893). This constitutes the first reported crystals of AT bound to an antibody. The monoclinic crystals belonged to space group P2₁, with unit-cell parameters a=85.52, b=148.50, c=93.82 Å, β=99.82°. The diffraction of the crystals extended to 2.56 Å resolution. The asymmetric unit contained two MEDI4893 Fab-AT complexes. This corresponds to a crystal volume per protein weight (VM) of 2.3 Å3 Da(-1) and a solvent content of 47%. The three-dimensional structure of this complex will contribute to an understanding of the molecular basis of the interaction of MEDI4893 with AT. It will also shed light on the mechanism of action of this antibody, the current evaluation of which in the field of S. aureus-mediated diseases makes it a particularly interesting case study. Finally, this study will provide the three-dimensional structure of AT in a monomeric state for the first time.

  19. Preliminary X-ray crystallographic analysis of an engineered variant of human chimera-type galectin-3 with a shortened N-terminal domain.

    PubMed

    Flores-Ibarra, Andrea; Ruiz, Federico M; Vértesy, Sabine; André, Sabine; Gabius, Hans-Joachim; Romero, Antonio

    2015-02-01

    How lectins translate sugar-encoded information into cellular effects not only depends on glycan recognition. Other domains of the protein can contribute to the functional profile of a lectin. Human galectin-3 (Gal-3), an adhesion/growth-regulatory galectin, is composed of three different domains and is thus called a chimera-type protein. In addition to the carbohydrate-recognition domain, this lectin encompasses an N-terminal domain consisting of a peptide harbouring two phosphorylation sites and nine non-triple-helical collagen-like repeats. This region plays an as yet structurally undefined role in Gal-3 aggregation and ligand recognition. To date, crystallization of full-length Gal-3 has not been achieved. With the aim of providing structural insights into this modular organization, a Gal-3 variant was crystallized maintaining the terminal peptide and three of the nine collagen-like repeats. The crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 94.04, b = 97.96, c = 236.20 Å, and diffracted to a resolution of 3.3 Å.

  20. Engineering of human tracheal tissue with collagen-enforced poly-lactic-glycolic acid non-woven mesh: a preliminary study in nude mice.

    PubMed

    Wu, Wei; Feng, Xue; Mao, Tianqiu; Feng, Xinghua; Ouyang, Hong-Wei; Zhao, Guifang; Chen, Fulin

    2007-06-01

    The purpose of the current study is to fabricate tissue engineered trachea with poly-lactic-glycolic acid (PLGA) non-woven mesh enforced by collagen type I. PLGA fibres coated with collagen solution were put together and fabricated into the shape of a human trachea, after drying and cross-linking treatment, a non-woven mesh with "C" shape formed. Chondrocytes from sheep nasal septum cartilage were expanded in vitro and seeded into PLGA/collagen non-woven mesh in the density of 5.0 x 10(7)mL(-1). After 5 days of in vitro incubation, six Cell-PLGA/collagen composites were implanted subcutaneously into the back of 6 nude mice to prefabricate a tissue engineering trachea. Eight weeks later, the cartilage formation was observed by gross inspection and histological examination. Cartilage-like tissue in the shape of the initial PLGA/collagen scaffold had been regenerated successfully without obvious inflammatory response. The tissue engineered trachea cartilage consisted of evenly spaced lacunae embedded in matrix stained red with safranin-O staining. The amount of GAGs in tissue engineered trachea cartilage reached 71.42% of normal value in native cartilage. This study demonstrated that collagen-enforced PLGA non-woven mesh facilitated the adhesion and proliferation of chondrocytes, it also owned adequate mechanical strength to serve as an ideal scaffold for trachea tissue engineering without internal support.

  1. Overproduction, purification, crystallization and preliminary X-ray analysis of human Fe65-PTB2 in complex with the amyloid precursor protein intracellular domain

    SciTech Connect

    Radzimanowski, Jens; Beyreuther, Konrad; Sinning, Irmgard; Wild, Klemens

    2008-05-01

    Alzheimer’s disease is characterized by proteolytic processing of the amyloid precursor protein (APP), which releases the aggregation-prone amyloid-β (Aβ) peptide and liberates the intracellular domain (AICD) that interacts with various adaptor proteins. The crystallized AICD–Fe65-PTB2 complex is of central importance for APP translocation, nuclear signalling, processing and Aβ generation. Alzheimer’s disease is associated with typical brain deposits (senile plaques) that mainly contain the neurotoxic amyloid β peptide. This peptide results from proteolytic processing of the type I transmembrane protein amyloid precursor protein (APP). During this proteolytic pathway the APP intracellular domain (AICD) is released into the cytosol, where it associates with various adaptor proteins. The interaction of the AICD with the C-terminal phosphotyrosine-binding domain of Fe65 (Fe65-PTB2) regulates APP translocation, signalling and processing. Human AICD and Fe65-PTB2 have been cloned, overproduced and purified in large amounts in Escherichia coli. A complex of Fe65-PTB2 with the C-terminal 32 amino acids of the AICD gave well diffracting hexagonal crystals and data have been collected to 2.1 Å resolution. Initial phases obtained by the molecular-replacement method are of good quality and revealed well defined electron density for the substrate peptide.

  2. Crystallization and preliminary X-ray diffraction analysis of CfaE, the adhesive subunit of the CFA/I fimbriae from human enterotoxigenic Escherichia coli

    SciTech Connect

    Li, Yong-Fu; Poole, Steven; Rasulova, Fatima; Esser, Lothar; Savarino, Stephen J.; Xia, Di

    2006-02-01

    The adhesin CfaE of the CFA/I fimbriae from human enterotoxigenic E. coli has been crystallized. CfaE crystals diffracted X-rays to better than 2.4 Å and phasing was solved by the SIRAS method. Enterotoxigenic Escherichia coli (ETEC) represents a formidable food and waterborne diarrheal disease threat of global importance. The first step in ETEC pathogenesis is bacterial attachment to small-intestine epithelial cells via adhesive fimbriae, many of which are genetically related to the prototype colonization factor antigen I (CFA/I). The minor fimbrial subunit CfaE is required for initiation of CFA/I fimbrial assembly and mediates bacterial attachment to host cell-surface receptors. A donor-strand complemented variant of CfaE (dscCfaE) was expressed with a hexahistidine tag, purified to homogeneity and crystallized using the hanging-drop vapor-diffusion method. X-ray diffraction data sets were collected to 2.4 Å resolution for both native and derivatized crystals and showed the symmetry of space group P6{sub 2}22, with unit-cell parameters a = b = 142.9, c = 231.9 Å. Initial phases were derived from the SIRAS approach and electron density showed two molecules in the crystallographic asymmetric unit. Sequence assignments were aided by anomalous signals from the selenium of an SeMet-derivatized crystal and from S atoms of a native crystal.

  3. [Occurrence of MRSA in pigs and in humans involved in pig production--preliminary results of a study in the northwest of Germany].

    PubMed

    Meemken, D; Cuny, C; Witte, W; Eichler, U; Staudt, R; Blaha, T

    2008-04-01

    In 2007, 678 pigs of all age groups out of 347 different farms from Lower Saxony and Northrhine-Westphalia and 86 persons occupationally exposed to pigs were investigated for their nasal colonisation with methicillin-resistant Staphylococcus areus (MRSA) by the Field Station for Epidemiology of the University of Veterinary Medicine Hannover and the Robert Koch-Institute. At the individual animal level, a frequency of positive results of 13% (n = 85 positive animals) and at the herd level, a frequency of positive results of 18% (n = 62 positive herds) were found. All isolates were assigned to the Multilocus Sequence Typing Type ST398. Within MRSA-positive herds, there were more MRSA-negative than MRSA-positive animals. Among the occupationally exposed persons (veterinarians, laboratory personnel and meat inspection personnel), 20 persons (23%) showed a nasal colonisation with MRSA ST398. A quite strong association between the intensity of the contact to pigs with the frequency of nasal colonisation in the occupationally exposed persons was detected. None of the animals or the humans nasally colonised by MRSA ST398 showed any clinical symptoms of a staphylococcal infection. Conclusions are drawn on the herd and intra-herd prevalence of the nasal colonisation of pigs with MRSA ST398 in pigs, but especially on which questions need to be addressed by further research.

  4. Purification, crystallization and preliminary crystallographic studies of C-terminal RNA recognition motif (RRM-3) of human ELAV-type RNA-binding protein 3 (ETR-3)

    PubMed Central

    Kashyap, Maruthi; Sharma, Ashwani; Bhavesh, Neel Sarovar

    2013-01-01

    Human embryonically lethal abnormal vision (ELAV)-type RNA-binding protein 3 (ETR-3) has been implicated in many aspects of RNA-processing events including alternative splicing, stability, editing and translation. RNA recognition motif 3 (RRM-3) is an independent C-terminal RNA-binding domain of ETR-3 that preferentially binds to UG-rich repeats of the nuclear or cytoplasmic pre-mRNA, and along with the other domains mediates the inclusion of cardiac troponin T (c-TNT) exon 5 in embryonic muscle, which is otherwise excluded in the adult. In the present study, RRM-3 was cloned, overexpressed, purified and crystallized by the hanging-drop vapour-diffusion method. The crystals diffracted to 3 Å resolution at the home source and belonged to space group P213, with unit-cell parameters a = b = c = 118.5 Å, α = β = γ = 90°. There were two molecules of RRM-3 in the asymmetric unit and the calculated Matthews coefficient (V M) was 6.35 Å3 Da−1, with a solvent content of 80.62%. Initial phases were determined by molecular replacement. PMID:24100559

  5. Human consumption as an invasive species management strategy. A preliminary assessment of the marketing potential of invasive Asian carp in the US.

    PubMed

    Varble, Sarah; Secchi, Silvia

    2013-06-01

    Over the past 20 years, Asian carp have invaded rivers and lakes in the Midwest and southern United States, with large negative impacts, such as encroachment on the habitat of native fish and mass die-off. They also respond to boat motors by jumping out of the water, which can cause harm to boaters and fishermen. Policymakers in the Great Lakes region between the US and Canada are worried about possible expansion of the Asian carp to their region and its effects on their fishing industry. A potential solution to the problem is to harvest Asian carp for human consumption. This study analyzes the results of the first national survey on the attitudes of US fish consumers towards Asian carp. We find that this is a potentially promising strategy. Most respondents would be willing to try a free sample of Asian carp and would be willing to pay for it. Because of the negative connotation attached to carp in general, this figure is encouraging. Creating demand for Asian carp could be a market based, cost-effective solution for a problem (invasive species) that is typically dealt with through command and control policies, if it is coupled with appropriate policies and safeguards to ensure the fish is eventually eradicated and not cultivated for profit after removal from US rivers and lakes.

  6. Can oxidative DNA damage be used as a biomarker of cancer risk in humans? Problems, resolutions and preliminary results from nutritional supplementation studies.

    PubMed

    Halliwell, B

    1998-12-01

    Damage to DNA by oxygen radicals and other reactive oxygen/nitrogen/chlorine species occurs in vivo despite the presence of multiple antioxidant defence and repair systems. Such damage is thought to make a significant contribution to the age-related development of cancer. Modulation of oxidative DNA damage by diet thus constitutes a "biomarker" putatively predictive of the effect of diet on cancer incidence, provided that DN