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Bacterial population detection is based on detection of adenosine triphosphate (ATP), chemical present in all living matter. Amount of ATP in sample, after chemically removing all nonbacterial ATP, is directly related to bacterial population. Sensitivity is improved by concentration step; specificity is improved by lowering pH of solution.
Cell and animal studies conducted onboard the International Space Station and formerly on Shuttle flights have provided groundbreaking data illuminating the deleterious biological response of bone to mechanical unloading. However the intercellular communicative mechanisms associated with the regulation of bone synthesis and bone resorption cells are still largely unknown. Connexin-43 (CX43), a gap junction protein, is hypothesized to play a significant role in osteoblast and osteocyte signaling. The purpose of this investigation was to evaluate within a novel three-dimensional microenvironment how the osteocyte-osteoblast gap-junction expression changes when cultures are exposed to exaggerated mechanical load. MLO-Y4 osteocyte-like cells were cultured on a 3D-Biotek polystyrene insert and placed in direct contact with an MC3T3-E1 pre-osteoblast co-cultured monolayer and exposed to 48 h of mechanical stimulation (pulsatile fluid flow (PFF) or monolayer cyclic stretch (MCS)) then evaluated for viability, proliferation, metabolism, and CX43 expression. Mono-cultured MLO-Y4 and MC3T3-E1 control experiments were conducted under PFF and MCS stimulation to observe how strain application stimuli (PFF cell membrane shear or MCS cell focal adhesion/attachment loading) initiates different signaling pathways or downstream regulatory controls. TotalLive cell count, viability and metabolic reduction (Trypan Blue, LIVEDead and Alamar Blue analysis respectively) indicate that mechanical activation of MC3T3-E1 cells inhibits proliferation while maintaining an average 1.04E4 reductioncell metabolic rate, *p0.05 n4. MLO-Y4s in monolayer culture increase in number when exposed to MCS loading but the percent of live cells within the population is low (46.3 total count, *p0.05 n4), these results may indicate an apoptotic signaling cascade. PFF stimulation of the three-dimensional co-cultures elicits a universal increase in CX43 in MLO-Y4 and MC3T3-E1 cells, illustrated by immunohistological observation. Increased CX43 expression is also observed with the three-dimensional co-cultures with MC3T3-E1 MCS stimulation but the increased gap-junction protein presence was limited to the osteoblast-osteocyte interface region. Previously reported PCR evaluation of osteogenic markers further corroborate that the co-cultured populations communicative networks play a role in translating mechanical signals to molecular messaging. These findings suggests an osteocyte-osteoblast gap-junction signaling feedback mechanism may regulate mechanotransduction of apoptosis initiation and transcription of cytokine signaling proteins responsible for stem cell niche recruitment much more directly than previously believed.
Cell and animal studies conducted onboard the International Space Station and formerly on Shuttle flights have provided groundbreaking data illuminating the deleterious biological response of bone to mechanical unloading. However the intercellular communicative mechanisms associated with the regulation of bone synthesis and bone resorption cells are still largely unknown. Connexin-43 (CX43), a gap junction protein, is hypothesized to play a significant role in osteoblast and osteocyte signaling. The purpose of this investigation was to evaluate within a novel three-dimensional microenvironment how the osteocyte-osteoblast gap-junction expression changes when cultures are exposed to exaggerated mechanical load. MLO-Y4 osteocyte-like cells were cultured on a 3D-Biotek polystyrene insert and placed in direct contact with an MC3T3-E1 pre-osteoblast co-cultured monolayer and exposed to 48 h of mechanical stimulation (pulsatile fluid flow (PFF) or monolayer cyclic stretch (MCS)) then evaluated for viability, proliferation, metabolism, and CX43 expression. Mono-cultured MLO-Y4 and MC3T3-E1 control experiments were conducted under PFF and MCS stimulation to observe how strain application stimuli (PFF cell membrane shear or MCS cell focal adhesionattachment loading) initiates different signaling pathways or downstream regulatory controls. TotalLive cell count, viability and metabolic reduction (Trypan Blue, LIVEDead and Alamar Blue analysis respectively) indicate that mechanical activation of MC3T3-E1 cells inhibits proliferation while maintaining an average 1.04E4 reductioncell metabolic rate, *p0.05 n4. MLO-Y4s in monolayer culture increase in number when exposed to MCS loading but the percent of live cells within the population is low (46.3 total count, *p0.05 n4), these results may indicate an apoptotic signaling cascade. PFF stimulation of the three-dimensional co-cultures elicits a universal increase in CX43 in MLO-Y4 and MC3T3-E1 cells, illustrated by immunohistological observation. Increased CX43 expression is also observed with the three-dimensional co-cultures with MC3T3-E1 MCS stimulation but the increased gap-junction protein presence was limited to the osteoblast-osteocyte interface region. Previously reported PCR evaluation of osteogenic markers further corroborate that the co-cultured populations communicative networks play a role in translating mechanical signals to molecular messaging. These findings suggests an osteocyte-osteoblast gap-junction signaling feedback mechanism may regulate mechanotransduction of apoptosis initiation and transcription of cytokine signaling proteins responsible for stem cell niche recruitment much more directly than previously believed.
We present an analysis of Galactic bulge stars from Hubble Space Telescope Wide Field Camera 3 observations of the Stanek window (l, b = [0.25, −2.15]) from two epochs approximately two years apart. This data set is adjacent to the provisional Wide-field Infrared Survey Telescope (WFIRST) microlensing field. Proper motions are measured for approximately 115,000 stars down to 28th mag in V band and 25th mag in I band, with accuracies of 0.5 mas/yr (20 km/s) at I ≈ 21. A cut on the longitudinal proper motion μl allows us to separate disk and bulge populations and produce bulge-only star counts that are corrected for photometric completeness and efficiency of the proper-motion cut. The kinematic dispersions and surface density in the field are compared to the nearby SWEEPS sight line, finding a marginally larger-than-expected gradient in stellar density. The observed bulge star counts and kinematics are further compared to the Besançon, Galaxia, and GalMod Galactic population synthesis models. We find that most of the models underpredict low-mass bulge stars by ∼33% below the main-sequence turnoff, and upwards of ∼70% at redder J and H wavebands. While considering inaccuracies in the Galactic models, we give implications for the exoplanet yield from the WFIRST microlensing mission.
Conventional methods for detecting indicator and pathogenic bacteria in water may underestimate the actual population due to sublethal environmental injury, inability of the target bacteria to take up nutrients and other physiological factors which reduce bacterial culturability. Rapid and direct methods are needed to more accurately detect and enumerate active bacteria. Such a methodological advance would provide greater sensitivity in assessing the microbiological safety of water and food. The principle goal of this presentation is to describe novel approaches we have formulated for the rapid and simultaneous detection of bacteria plus the determination of their physiological activity in water and other environmental samples. The present version of our method involves the concentration of organisms by membrane filtration or immunomagnetic separation and combines an intracellular fluorochrome (CTC) for assessment of respiratory activity plus fluorescent-labelled antibody detection of specific bacteria. This approach has also been successfully used to demonstrate spatial and temporal heterogeneities of physiological activities in biofilms when coupled with cryosectioning. Candidate physiological stains include those capable of determining respiratory activity, membrane potential, membrane integrity, growth rate and cellular enzymatic activities. Results obtained thus far indicate that immunomagnetic separation can provide a high degree of sensitivity in the recovery of seeded target bacteria (Escherichia coli O157:H7) in water and hamburger. The captured and stained target bacteria are then enumerated by either conventional fluorescence microscopy or ChemScan(R), a new instrument that is very sensitive and rapid. The ChemScan(R) laser scanning instrument (Chemunex, Paris, France) provides the detection of individual fluorescently labelled bacterial cells using three emission channels in less than 5 min. A high degree of correlation has been demonstrated between results obtained with the ChemScan and traditional plate counts of mixed natural bacterial populations in water. The continuing evolution of these methods will be valuable in the rapid and accurate analysis of environmental samples.
Models of the faint galaxy contribution to the diffuse extragalactic background light are presented, which are consistent with current data on faint galaxy number counts and redshifts. The autocorrelation function of surface brightness fluctuations in the extragalactic diffuse light is predicted, and the way in which these predictions depend on the cosmological model and assumptions of biasing is determined. It is confirmed that the recent deep infrared number counts are most compatible with a high density universe (Omega-0 is approximately equal to 1) and that the steep blue counts then require an extra population of rapidly evolving blue galaxies. The faintest presently detectable galaxies produce an interesting contribution to the extragalactic diffuse light, and still fainter galaxies may also produce a significant contribution. These faint galaxies still only produce a small fraction of the total optical diffuse background light, but on scales of a few arcminutes to a few degrees, they produce a substantial fraction of the fluctuations in the diffuse light.
The purpose of this investigation was to examine microbial communities of simulated wastewater effluent from hollow fiber membrane bioreactors collected from the Space Life Science Laboratory and Texas Technical University. Microbes were characterized using quantitative polymerase chain reaction where a total count of bacteria and fungi were determined. The primers that were used to determine the total count of bacteria and fungi were targeted for 16S rDNA genes and the internal transcribed spacer, respectively. PCR products were detected with SYBR Green I fluorescent dye and a melting curve analysis was performed to identify unique melt profiles resulting from DNA sequence variations from each species of the community. Results from both the total bacteria and total fungi count assays showed that distinct populations were present in isolates from these bioreactors. This was exhibited by variation in the number of peaks observed on the melting curve analysis graph. Further analysis of these results using species-specific primers will shed light on exactly which microbes are present in these effluents. Information gained from this study will enable the design of a system that can efficiently monitor microbes that play a role in the biogeochemical cycling of nitrogen in wastewater on the International Space Station to assist in the design of a sustainable system capable of converting this nutrient.
We present 0.5-2 keV, 2-8 keV, 4-8 keV, and 0.5-8 keV (hereafter soft, hard, ultra-hard, and full bands, respectively) cumulative and differential number-count (log N-log S ) measurements for the recently completed approx. equal to 4 Ms Chandra Deep Field-South (CDF-S) survey, the deepest X-ray survey to date. We implement a new Bayesian approach, which allows reliable calculation of number counts down to flux limits that are factors of approx. equal to 1.9-4.3 times fainter than the previously deepest number-count investigations. In the soft band (SB), the most sensitive bandpass in our analysis, the approx. equal to 4 Ms CDF-S reaches a maximum source density of approx. equal to 27,800 deg(sup -2). By virtue of the exquisite X-ray and multiwavelength data available in the CDF-S, we are able to measure the number counts from a variety of source populations (active galactic nuclei (AGNs), normal galaxies, and Galactic stars) and subpopulations (as a function of redshift, AGN absorption, luminosity, and galaxy morphology) and test models that describe their evolution. We find that AGNs still dominate the X-ray number counts down to the faintest flux levels for all bands and reach a limiting SB source density of approx. equal to 14,900 deg(sup -2), the highest reliable AGN source density measured at any wavelength. We find that the normal-galaxy counts rise rapidly near the flux limits and, at the limiting SB flux, reach source densities of approx. equal to 12,700 deg(sup -2) and make up 46% plus or minus 5% of the total number counts. The rapid rise of the galaxy counts toward faint fluxes, as well as significant normal-galaxy contributions to the overall number counts, indicates that normal galaxies will overtake AGNs just below the approx. equal to 4 Ms SB flux limit and will provide a numerically significant new X-ray source population in future surveys that reach below the approx. equal to 4 Ms sensitivity limit. We show that a future approx. equal to 10 Ms CDF-S would allow for a significant increase in X-ray-detected sources, with many of the new sources being cosmologically distant (z greater than or approx. equal to 0.6) normal galaxies.
The major goal of part I of this study was to compare varying doses and dose rates of whole-body gamma-radiation on lymphoid cells and organs. C57BL/6 mice (n = 75) were exposed to 0, 0.5, 1.5, and 3.0 Gy gamma-rays (60Co) at 1 cGy/min (low-dose rate, LDR) and 80 cGy/min (high-dose rate, HDR) and euthanized 4 days later. A significant dose-dependent loss of spleen mass was observed with both LDR and HDR irradiation; for the thymus this was true only with HDR. Decreasing leukocyte and lymphocyte numbers occurred with increasing dose in blood and spleen at both dose rates. The numbers (not percentages) of CD3+ T lymphocytes decreased in the blood in a dose-dependent manner at both HDR and LDR. Splenic T cell counts decreased with dose only in HDR groups; percentages increased with dose at both dose rates. Dose-dependent decreases occurred in CD4+ T helper and CD8+ T cytotoxic cell counts at HDR and LDR. In the blood the percentages of CD4+ cells increased with increasing dose at both dose rates, whereas in the spleen the counts decreased only in the HDR groups. The percentages of the CD8+ population remained stable in both blood and spleen. CD19+ B cell counts and percentages in both compartments declined markedly with increasing HDR and LDR radiation. NK1.1+ natural killer cell numbers and proportions remained relatively stable. Overall, these data indicate that the observed changes were highly dependent on the dose, but not dose rate, and that cells in the spleen are more affected by dose rate than those in blood. The results also suggest that the response of lymphocytes in different body compartments may be variable.
Optical-infrared observations are used to show that the sub-mJy sources, discovered in deep radio surveys and responsible for the upturn in the radio source counts at sub-mJy levels, represent a population of star-forming galaxies at intermediate redshifts and with Mv between about -23 and -20. The very high frequency of line-emission, the very large incidence of galaxy mergers and interactions associated with these sub-mJy sources, and the good agreement between the sub-mJy radio source counts and the counts of starburst galaxies from deep IRAS surveys imply that these star-forming galaxies are in fact undergoing starbursts.
The fine scale mixing produced by a delta tab in a shear layer has been studied experimentally. The tab was placed at the trailing edge of a splitter plate which produced a turbulent two-stream mixing layer. The tab apex tilted downstream and into the high speed stream. Hot-wire measurements in the 3-D space behind the tab detailed the three velocity components as well as the small scale population distributions. These small scale eddies, which represent the peak in the dissipation spectrum, were identified and counted using the Peak-Valley-Counting technique. It was found that the small scale populations were greater in the shear region behind the tab, with the greatest increase occurring where the shear layer underwent a sharp turn. This location was near, but not coincident, with the core of the streamwise vortex, and away from the region exhibiting maximum turbulence intensity. Moreover, the tab increased the most probably frequency and strain rate of the small scales. It made the small scales smaller and more energetic.
Fecundity, the number of young produced by a breeding pair during a breeding season, is a primary component in evolutionary and ecological theory and applications. Fecundity can be influenced by many environmental factors and requires long-term study due to the range of variation in ecosystem dynamics. Fecundity data often include a large proportion of zeros when many pairs fail to produce any young during a breeding season due to nest failure or when all young die independently after fledging. We conducted color banding and monthly censuses of Florida scrub-jays (Aphelocoma coerulescens) across 31 years, 15 populations, and 761 territories along central Florida’s Atlantic coast. We quantified how fecundity (juveniles/pair-year) was influenced by habitat quality, presence/absence of nonbreeders, population density, breeder experience, and rainfall, with a zero-inflated Bayesian hierarchical model including both a Bernoulli (e.g., brood success) and a Poisson (counts of young) submodel, and random effects for year, population, and territory. The results identified the importance of increasing “strong” quality habitat, which was a mid-successional state related to fire frequency and extent, because strong territories, and the proportion of strong territories in the overall population, influenced fecundity of breeding pairs. Populations subject to supplementary feeding also had greater fecundity. Territory size, population density, breeder experience, and rainfall surprisingly had no or small effects. Different mechanisms appeared to cause annual variation in fecundity, as estimates of random effects were not correlated between the success and count submodels. The increased fecundity for pairs with nonbreeders, compared to pairs without, identified empirical research needed to understand how the proportion of low-quality habitats influences population recovery and sustainability, because dispersal into low-quality habitats can drain nonbreeders from strong territories and decrease overall fecundity. We also describe how long term study resulted in reversals in our understanding because of complications involving habitat quality, sociobiology, and population density.
We experienced much success in reaching our stated goals in our original MDAP proposal. Our work made substantial contributions towards an integrated understanding of the counting and calibration of crater data on Mars, and changing nature of the Martian surface influenced by craters, water, and wind, and their general relationship to Martian geothermal history. We accomplished this while being to responsive to the rapid changes in the field brought about by several key NASA missions that returned data during the life of the grant. Our integrated effort included three stages: The first major area of research (Crater Count Research) was conducted by Jennifer Grier (P.I.), Lazslo Keszthelyi (Collaborator), William Hartmann (Collaborator), with assistance from Dan Berman (Graduate student) and concerned the mapping and the collection of crater count data on various Martian terrains. The second major area of study (Absolute Age Calibration) was conducted by William Bottke (Co-I) at SWRI, and concerned constraining the nature of the Moon and Mars impactor populations to create better absolute age calibrations for counted areas. The third major area of study was the integration and leverage of this effort with ongoing related Mars crater work at PSI (Integrated and Continuing Studies - Older Volcanoes), headed by David Crown (PSI Scientist), assisted by Les Bleamaster (PSI Scientist) and Dan Berman (Graduate Student).
The species of sparrow identified by foraging under dense thickets, the Eastern Towhee (Pipilo erythrophthalmus) resides in the Eastern United States, some populations either permanently living in the southeastern US or migratory populations living in the North. The mostly migratory Eastern Towhee has been experiencing population changes since the 1980s in the Eastern Seaboard. This species is increasing during the winter and decreasing during the summer, which is their breeding season. The Christmas Bird Count shows that in the winter, the Eastern Towhee populations have been increasing in their northern wintering range by 0.2625% in Maryland, 0.056% in West Virginia, 0.133% in New Jersey, 0.049% in Virginia, and 0.224% in Delaware from 1984-2019. The Breeding Bird Survey Data shows that in the summer, the Eastern Towhee populations have been declining in the Eastern United States by -4.0635% in Maryland, -3.073% in West Virginia, -15.0722% in New Jersey, -3.9375% in Virginia, and -6.3735% in Delaware from 1984-2015. The average climate anomalies in June range from -0.108 to 0.3705 degrees Celsius while the average climate anomalies in January range from 0.1395 to 0.5014 degrees Celsius. The Eastern Towhee are moving northward towards their northern wintering range during the winter and we explore the hypothesis of northward range shift in association with climate change. Our research displays how the behavior patterns of species are varying due to this stressor and how to better conserve species like the Eastern Towhee.
Interplanetary dust is interesting both from a physical point of view and from the awareness that the population poses a substantial collision risk to manned and unmanned spacecraft alike. Insight into the time evolution of the population gives important insights into solar system formation and, we argue, will offer clues as to the behavior of solar cycles predating modern ways of viewing the sun (e.g., sun spot counting, magnetograms, etc.). In addition, knowing how the dust population moves under the variety of forces each grain experiences will inform and better prepare manned and robotic spacecraft missions to mitigate the risk posed by dust. With these aims in mind, we developed a simple dynamical model for dust motion based on the work of Czechowski and Mann [1]. A byproduct of that development is a clear analytic model that demonstrates how the interplay of the Poynting-Robertson and Lorenz perturbation forces on the Keplerian motion of charged dust grains around the Sun can result in dust populations that move inward, outward, or remain confined in annular rings about the Sun. The timing of any of these behaviors is strongly dependent on the physical properties of the grain (mass, charge and reflectivity) and the evolution of the Sun’s dipole field.
A disc and halo population model is constructed to fit star counts and color data down to V approximately 23 at absolute value of b = 90 deg. This model is used to predict star counts and colors down to V approximately 30. Deviations from these extrapolated relationships provide constraints on the number of faint quasars and black dwarf stars. It is shown that extra-galactic globular clusters start contributing significantly to star counts at V approximately 25 and are more numerous than stars for V 31. Morphological studies of galaxies with approximately 0.5, were made with the space telescope. Significant constraints on theoretical models that describe the evolution of clusters of galaxies are provided.
Results from the Hawaii imaging and spectroscopy deep survey are discussed. Cumulative galaxy counts now exceed 10 exp 6/deg. To B = 24 the bulk of these galaxies lie at low redshift. Flat spectrum sources constitute a fraction of about 30-40 percent of the sources at B = 24. There are not enough of these to explain the high number counts, but they are an exceedingly interesting population in their own right. It is argued that they are unlikely to lie at z of about 1, but could be either massive starbursters at z of about 2 - 3 or dwarf starbursting galaxies at very low redshift (less than approximately 0.1). In either case they constitute a major episode in galaxy formation.
Pinholes or other defect sites in a protective oxide coating provide pathways for atomic oxygen in low Earth orbit to reach underlying material. One concept for enhancing the lifetime of materials in low Earth orbits is to apply a leveling coating to the material prior to applying any reflective and protective coatings. Using a surface tension leveling coating concept, a low viscosity epoxy was applied to the surface of several composite coupons. A protective layer of 1000 A of SiO2 was deposited on top of the leveling coating, and the coupons were exposed to an atomic oxygen environment in a plasma asher. Pinhole populations per unit area were estimated by counting the number of undercut sites observed by scanning electron microscopy. Defect density values of 180,000 defects/sq cm were reduced to about 1000 defects/sq cm as a result of the applied leveling coating. These improvements occur at a mass penalty of about 2.5 mg/sq cm.