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At least 289 records · Page 16

Tubulinlike protein from Spirochaeta bajacaliforniensis

Tubulin proteins are the fundamental subunits of all polymeric microtubule-based eukaryotic structures. Long, hollow structures each composed of 13 protofilaments as revealed by electron microscopy, microtubules (240 angstroms in diameter) are nearly ubiquitous in eukaryotes. These proteins have been the subject of intense biochemical and biophysical interest since the early 1970s and are of evolutionary interest as well. If tubulin-based structures (i.e., neurotubules, mitotic spindle tubules, centrioles, kinetosomes, axonemes, etc.) evolved from spirochetes by way of motility symbioses, tubulin homologies with spirochete proteins should be detectable. Tubulin proteins are widely thought to be limited to eukaryotes. Yet both azotobacters and spirochetes have shown immunological cross-reactivity with anitubulin antibodies. In neither of these studies was tubulin isolated nor any specific antigen identified as responsible for the immunoreactivity. Furthermore, although far less uniform in structure than eukaryotic microtubules, various cytoplasmic fibers and tubules (as seen by electron microscopy) have been reported in several types of prokaryotes (e.g., Spirochaeta; large termite spirochetes; treponemes; cyanobacteria; and Azotobacter. This work forms a part of our long-range study of the possible prokaryotic origin of tubulin and microtubules. Spirochetes are helically shaped gram-negative motile prokaryotes. They differ from all other bacteria in that the position of their flagella is periplasmic: their flagella lie between the inner and outer membranes of the gram-negative cell wall. Some of the largest spirochetes have longitudinally aligned 240 angstroms microtubules. Unfortunately, in spite of many attempts, all of the larger spirochetes (family Pillotaceae) with well-defined cytoplasmic tubules and antitubulin immunoreactivity are not cultivable. However, a newly described spirochete species (Spirochaeta bajacaliforniensis) possessing cytoplasmic fibers displays antitubulin immunoreactivity in whole-cell preparations. Since preliminary observations suggested that Spirochaeta bajacaliforniensis proteins may be related to eukaryotic tubulins, their characterization was undertaken. Brain tubulin can be purified by utilizing its ability to polymerize at warm temperatures and to depolymerize in the cold. After several cycles of sedimentation and redissolution the microtubule fraction is composed of 75% tubulin and 20% high molecular mass microtubule-associated proteins (MAPs). In this paper we report that components of cell lysates, prepared from a spirochete that contains cytoplasmic fibers (Spirochaeta bajacaliforniensis), also exhibit the property of temperature-dependent cyclical sedimentation. Additionally we report the identification and characterization of the polypeptide responsible for cross-reactivity with antitubulin antiserum.

Non-NASA Center↗

Utilization of ammonium as a nitrogen source: effects of ambient acidity on growth and nitrogen accumulation by soybean

Dry matter accumulation of plants utilizing NH4+ as the sole nitrogen source generally is less than that of plants receiving NO3- unless acidity of the root-zone is controlled at a pH of about 6.0. To test the hypothesis that the reduction in growth is a consequence of nitrogen stress within the plant in response to effects of increased acidity during uptake of NH4+ by roots, nonnodulated soybean plants (Glycine max [L.] Merr. cv Ransom) were grown for 24 days in flowing nutrient culture containing 1.0 millimolar NH4+ as the nitrogen source. Acidities of the culture solutions were controlled at pH 6.1, 5.1, and 4.1 +/- 0.1 by automatic additions of 0.01 N H2SO4 or Ca(OH)2. Plants were sampled at intervals of 3 to 4 days for determination of dry matter and nitrogen accumulation. Rates of NH4+ uptake per gram root dry weight were calculated from these data. Net CO2 exchange rates per unit leaf area were measured on attached leaves by infrared gas analysis. When acidity of the culture solution was increased from pH 6.1 to 5.1, dry matter and nitrogen accumulation were reduced by about 40% within 14 days. Net CO2 exchange rates per unit leaf area, however, were not affected, and the decreased growth was associated with a reduction in rates of appearance and expansion of new leaves. The uptake rates of NH4+ per gram root were about 25% lower throughout the 24 days at pH 5.1 than at 6.1. A further increase in solution acidity from pH 5.1 to 4.1 resulted in cessation of net dry matter production and appearance of new leaves within 10 days. Net CO2 exchange rates per unit leaf area declined rapidly until all viable leaves had abscised by 18 days. Uptake rates of NH4+, which were initially about 50% lower at pH 4.1 than at 6.1 continued to decline with time of exposure until net uptake ceased at 10 days. Since these responses also are characteristic of the sequence of responses that occur during onset and progression of a nitrogen stress, they corroborate our hypothesis.

NASA Discipline Number 61-10↗

Cyclic variations in nitrogen uptake rate of soybean plants: effects of external nitrate concentration

Net uptake of NO3- by non-nodulated soybean plants [Glycine max (L.) Merr. cv. Ransom] growing in flowing hydroponic cultures containing 0.5, 1.0 and 10.0 mol m-3 NO3- was measured daily during a 24-d period of vegetative development to determine if amplitude of maximum and minimum rates of net NO3- uptake are responsive to external concentrations of NO3-. Removal of NO3- from the replenished solutions during each 24-h period was determined by ion chromatography. Neither dry matter accumulation nor the periodicity of oscillations in net uptake rate was altered by the external NO3- concentrations. The maxima of the oscillations in net uptake rate, however, increased nearly 3-fold in response to external NO3- concentrations. The maxima and minima, respectively, changed from 4.0 and 0.6 mmol NO3- per gram root dry weight per day at an external solution level of 0.5 mol m-3 NO3- to 15.2 and -2.7 mmol NO3- per gram root dry weight per day at an external solution level of 10.0 mol m-3 NO3-. The negative values for minimum net uptake rate from 10.0 mol m-3 NO3- solutions show that net efflux was occurring and indicate that the magnitude of the efflux component of net uptake was responsive to external concentration of NO3-.

Non-NASA Center↗

Thermal Performance of a Multi-Evaporator Loop Heat Pipe with Thermal Masses and Thermoelectric Coolers

This paper describes thermal performance of a loop heat pipe (LHP) with two evaporators and two condensers in ambient testing. Each evaporator has an outer diameter of 15mm and a length of 76mm, and has an integral compensation chamber (CC). An aluminum mass of 500 grams is attached to each evaporator to simulate the instrument mass. A thermoelectric cooler (TEC) is installed on each CC to provide heating as well as cooling for CC temperature control. A flow regulator is installed in the condenser section to prevent vapor from going back to the evaporators in the event that one of the condensers is fully utilized. Ammonia was used as the working fluid. Tests conducted included start-up, power cycle, heat load sharing, sink temperature cycle, operating temperature control with TECs, and capillary limit tests. Experimental data showed that the loop could start with a heat load of less than 10W even with added thermal masses. The loop operated stably with even and uneven evaporator heat loads, and even and uneven condenser sink temperatures. The operating temperature could be controlled within +/- 0.5K of the set point temperature using either or both TECs, and the required TEC control heater power was less than 2W under most test conditions. Heat load sharing between the two evaporators was also successfully demonstrated. The loop had a heat transport capability of 120W to 140W, and could recover from a dry-out when the heat load was reduced. The 500-gram aluminum mass on each evaporator had a negligible effect on the loop operation. Existing LHPs servicing orbiting spacecraft have a single evaporator with an outer diameter of about 25mm. Important performance characteristics demonstrated by this LHP included: 1) Operation of an LHP with 15mm diameter evaporators; 2) Robustness and reliability of an LHP with multiple evaporators and multiple condensers under various test conditions; 3) Heat load sharing among LHP evaporators; 4) Effectiveness of TECs in controlling the LHP operating temperature; and 5 ) Effectiveness of the flow regulator in preventing vapor from going back the evaporators.

Ku, Jen-Tung↗

Determining divergence times with a protein clock: update and reevaluation

A recent study of the divergence times of the major groups of organisms as gauged by amino acid sequence comparison has been expanded and the data have been reanalyzed with a distance measure that corrects for both constraints on amino acid interchange and variation in substitution rate at different sites. Beyond that, the availability of complete genome sequences for several eubacteria and an archaebacterium has had a great impact on the interpretation of certain aspects of the data. Thus, the majority of the archaebacterial sequences are not consistent with currently accepted views of the Tree of Life which cluster the archaebacteria with eukaryotes. Instead, they are either outliers or mixed in with eubacterial orthologs. The simplest resolution of the problem is to postulate that many of these sequences were carried into eukaryotes by early eubacterial endosymbionts about 2 billion years ago, only very shortly after or even coincident with the divergence of eukaryotes and archaebacteria. The strong resemblances of these same enzymes among the major eubacterial groups suggest that the cyanobacteria and Gram-positive and Gram-negative eubacteria also diverged at about this same time, whereas the much greater differences between archaebacterial and eubacterial sequences indicate these two groups may have diverged between 3 and 4 billion years ago.

NASA Discipline Exobiology↗

Mechanosensitive channels of Escherichia coli: the MscL gene, protein, and activities

Although mechanosensory responses are ubiquitous and diverse, the molecular bases of mechanosensation in most cases remain mysterious MscL, a mechanosensitive channel of large conductance of Escherichia coli and its bacterial homologues are the first and currently only channel molecules shown to directly sense mechanical stretch of the membrane. In response to the tension conveyed via the lipid bilayer, MscL increases its open probability by several orders of magnitude. In the present review we describe the identification, cloning, and first sets of biophysical and structural data on this simplest mechanosensory molecule. We discovered a 2.5-ns mechanosensitive conductance in giant E. coli spheroplasts. Using chromatographies to enrich the target and patch clamp to assay the channel activity in liposome-reconstituted fractions, we identified the MscL protein and cloned the mscL gene. MscL comprises 136 amino acid residues (15 kDa), with two highly hydrophobic regions, and resides in the inner membrane of the bacterium. PhoA-fusion experiments indicate that the protein spans the membrane twice with both termini in the cytoplasm. Spectroscopic techniques show that it is highly helical. Expression of MscL tandems and covalent cross-linking suggest that the active channel complex is a homo-hexamer. We have identified several residues, which when deleted or substituted, affect channel kinetics or mechanosensitivity. Although unique when discovered, highly conserved MscL homologues in both gram-negative and gram-positive bacteria have been found, suggesting their ubiquitous importance among bacteria.

Non-NASA Center↗

Microbial monitoring of spacecraft and associated environments

Rapid microbial monitoring technologies are invaluable in assessing contamination of spacecraft and associated environments. Universal and widespread elements of microbial structure and chemistry are logical targets for assessing microbial burden. Several biomarkers such as ATP, LPS, and DNA (ribosomal or spore-specific), were targeted to quantify either total bioburden or specific types of microbial contamination. The findings of these assays were compared with conventional, culture-dependent methods. This review evaluates the applicability and efficacy of some of these methods in monitoring the microbial burden of spacecraft and associated environments. Samples were collected from the surfaces of spacecraft, from surfaces of assembly facilities, and from drinking water reservoirs aboard the International Space Station (ISS). Culture-dependent techniques found species of Bacillus to be dominant on these surfaces. In contrast, rapid, culture-independent techniques revealed the presence of many Gram-positive and Gram-negative microorganisms, as well as actinomycetes and fungi. These included both cultivable and noncultivable microbes, findings further confirmed by DNA-based microbial detection techniques. Although the ISS drinking water was devoid of cultivable microbes, molecular-based techniques retrieved DNA sequences of numerous opportunistic pathogens. Each of the methods tested in this study has its advantages, and by coupling two or more of these techniques even more reliable information as to microbial burden is rapidly obtained. Copyright 2004 Springer-Verlag.

Environmental Monitoring/methods↗

The Vapor Pressure of Palladium at Temperatures up to 1973K

Understanding high-temperature processes is imperative for modeling the formation of the solar system. It is unfortunate that since the 1950 s little has been done in the area of thermodynamics to continue gaining information on metals such as iron (Fe), nickel (Ni), cobalt (Co), palladium (Pd) and many others. Although the vapor pressures of these metals can be extrapolated to higher temperatures, the data is often limited to temperature ranges too low to be applicable to processes that occur during the formation of the solar system (T approx. 2000K). Experimental techniques inhibited the data in the past by restricting the testing of metals to temperatures below their melting point. Today, higher temperature testing is possible by using a Thermo- Cahn Thermogravimetric system that is able to reach temperatures up to 1973K in vacuo and measure a 10 gram change in a sample with mass of up to 100 grams.

Gardner, K. G.↗

Spoken Language Processing in the Clarissa Procedure Browser

Clarissa, an experimental voice enabled procedure browser that has recently been deployed on the International Space Station, is as far as we know the first spoken dialog system in space. We describe the objectives of the Clarissa project and the system's architecture. In particular, we focus on three key problems: grammar-based speech recognition using the Regulus toolkit; methods for open mic speech recognition; and robust side-effect free dialogue management for handling undos, corrections and confirmations. We first describe the grammar-based recogniser we have build using Regulus, and report experiments where we compare it against a class N-gram recogniser trained off the same 3297 utterance dataset. We obtained a 15% relative improvement in WER and a 37% improvement in semantic error rate. The grammar-based recogniser moreover outperforms the class N-gram version for utterances of all lengths from 1 to 9 words inclusive. The central problem in building an open-mic speech recognition system is being able to distinguish between commands directed at the system, and other material (cross-talk), which should be rejected. Most spoken dialogue systems make the accept/reject decision by applying a threshold to the recognition confidence score. NASA shows how a simple and general method, based on standard approaches to document classification using Support Vector Machines, can give substantially better performance, and report experiments showing a relative reduction in the task-level error rate by about 25% compared to the baseline confidence threshold method. Finally, we describe a general side-effect free dialogue management architecture that we have implemented in Clarissa, which extends the "update semantics'' framework by including task as well as dialogue information in the information state. We show that this enables elegant treatments of several dialogue management problems, including corrections, confirmations, querying of the environment, and regression testing.

Rayner, M.↗

GLOBAL REFERENCE ATMOSPHERIC MODELS FOR AEROASSIST APPLICATIONS

Aeroassist is a broad category of advanced transportation technology encompassing aerocapture, aerobraking, aeroentry, precision landing, hazard detection and avoidance, and aerogravity assist. The eight destinations in the Solar System with sufficient atmosphere to enable aeroassist technology are Venus, Earth, Mars, Jupiter, Saturn, Uranus, Neptune, and Saturn's moon Titan. Engineering-level atmospheric models for five of these targets - Earth, Mars, Titan, Neptune, and Venus - have been developed at NASA's Marshall Space Flight Center. These models are useful as tools in mission planning and systems analysis studies associated with aeroassist applications. The series of models is collectively named the Global Reference Atmospheric Model or GRAM series. An important capability of all the models in the GRAM series is their ability to simulate quasi-random perturbations for Monte Carlo analysis in developing guidance, navigation and control algorithms, for aerothermal design, and for other applications sensitive to atmospheric variability. Recent example applications are discussed.

Duvall, Aleta↗

Thermal performance of a multi-evaporator loop heat pipe with thermal masses and thermal electrical coolers

This paper describes thermal performance of a loop heat pipe (LHP) with two evaporators and two condensers in ambient testing. Each evaporator has an outer diameter of 15mm and a length of 76mm, and has an integral compensation chamber (CC). An aluminum mass of 500 grams is attached to each evaporator to simulate the instrument mass. A thermal electric cooler (TEC) is installed on each CC to provide heating as well as cooling for CC temperature control. A flow regulator is installed in the condenser section to prevent vapor from going back to the evaporators in the event that one of condenser is fully utilized. Ammonia was used ad the working fluid. Tests conducted included start-up, power cycle, heat load sharing, sink temperature cycle, operating temperature control with TECs, and capillary limit tests. Experimental data showed that the loop could start with a heat load of less than 1OW even with added thermal masses. The loop operated stably with even and uneven evaporator heat loads, and even and uneven condenser sink temperatures. The operating temperature could be controlled within +/-0.5K of the set point temperature using either or both TECs, and the required TEC control heater power was less than 2W under most test conditions. Heat load sharing between the two evaporators was also successfully demonstrated. The loop had a heat transport capability of 120W to 140W, and could recover from a dry-out when the heat load was reduced. The 500-gram aluminum mass on each evaporator had a negligible effect on the loop operation. Existing LHPs servicing the orbiting spacecraft have a single evaporator with an outer diameter of about 25mm. Important performance characteristics demonstrated by this LHP included: 1) Operation of an LHP with 15mm diameter evaporators; 2) Robustness and reliability of an LHP with multiple evaporators and multiple condensers under various test conditions; 3) Heat load sharing among LHP evaporators; 4) Effectiveness of TECs in controlling the LHP operating temperature; and 5) Effectiveness of the flow regulator in preventing vapor from going back the evaporators.

loop heat pipes↗

Investigation and Comparison between New Satellite Impact Test Results and NASA Standard Breakup Model

This paper summarizes two new satellite impact tests conducted in order to investigate on the outcome of low- and hyper-velocity impacts on two identical target satellites. The first experiment was performed at a low velocity of 1.5 km/s using a 40-gram aluminum alloy sphere, whereas the second experiment was performed at a hyper-velocity of 4.4 km/s using a 4-gram aluminum alloy sphere by two-stage light gas gun in Kyushu Institute of Technology. To date, approximately 1,500 fragments from each impact test have been collected for detailed analysis. Each piece was analyzed based on the method used in the NASA Standard Breakup Model 2000 revision. The detailed analysis will conclude: 1) the similarity in mass distribution of fragments between low and hyper-velocity impacts encourages the development of a general-purpose distribution model applicable for a wide impact velocity range, and 2) the difference in area-to-mass ratio distribution between the impact experiments and the NASA standard breakup model suggests to describe the area-to-mass ratio by a bi-normal distribution.

Sakuraba, K.↗

Status of Solar Sail Propulsion: Moving Toward an Interstellar Probe

NASA's In-Space Propulsion Technology Program has developed the first-generation of solar sail propulsion systems sufficient to accomplish inner solar system science and exploration missions. These first-generation solar sails, when operational, will range in size from 40 meters to well over 100 meters in diameter and have an areal density of less than 13 grams-per-square meter. A rigorous, multiyear technology development effort culminated last year in the testing of two different 20-meter solar sail systems under thermal vacuum conditions. This effort provided a number of significant insights into the optimal design and expected performance of solar sails as well as an understanding of the methods and costs of building and using them. In a separate effort, solar sail orbital analysis tools for mission design were developed and tested. Laboratory simulations of the effects of long-term space radiation exposure were also conducted on two candidate solar sail materials. Detailed radiation and charging environments were defined for mission trajectories outside the protection of the earth's magnetosphere, in the solar wind environment. These were used in other analytical tools to prove the adequacy of sail design features for accommodating the harsh space environment. Preceding, and in conjunction with these technology efforts, NASA sponsored several mission application studies for solar sails, including one that would use an evolved sail capability to support humanity's first mission into nearby interstellar space. The proposed mission is called the Interstellar Probe. The Interstellar Probe might be accomplished in several ways. A 200-meter sail, with an areal density approaching 1 gram-per-square meter, could accelerate a robotic probe to the very edge of the solar system in just under 20 years from launch. A sail using the technology just demonstrated could make the same mission, but take significantly longer. Conventional chemical propulsion systems would require even longer flight times. Spinner sails of the type being explored by the Japanese may also be a good option, but the level of maturity in that technology is not clear. While the technology to support a 200-meter, ultralightweight sail mission is not yet in hand, the recent NASA investments in solar sail technology are an essential first step toward making it a reality. This paper will describe the status of solar sail propulsion within NASA, near-term solar sail mission applications, and the plan to advance the technology to the point where the Interstellar Probe mission can be flown.

Johnson, Les↗

Fine Aerosol Bulk Composition Measured on WP-3D Research Aircraft in Vicinity of the Northeastern United States - Results from NEAQS

During the New England Air Quality Study (NEAQS) in the summer of 2004, airborne measurements were made of the major inorganic ions and the water-soluble organic carbon (WSOC) of the submicron (PM(sub 1.0)) aerosol. These and ancillary data are used to describe the overall aerosol chemical characteristics encountered during the study. Fine particle mass was estimated from particle volume and a calculated density based on measured particle composition. Fine particle organic matter (OM) was estimated from WSOC and a mass balance analysis. The aerosol over the northeastern United States (U.S.) and Canada was predominantly sulfate and associated ammonium, and organic components, although in unique plumes additional ionic components were also periodically above detection limits. In power generation regions, and especially in the Ohio River Valley region, the aerosol tended to be predominantly sulfate (approximately 60% micro gram /micro gram) and apparently acidic, based on an excess of measured anions compared to cations. In all other regions where sulfate concentrations were lower and a smaller fraction of overall mass, the cations and anions were balanced suggesting a more neutral aerosol. In contrast, the WSOC and estimated OM were more spatially uniform and the fraction of OM relative to PM mass was largely influenced by sources of sulfate. The study median OM mass fraction was 40%. Throughout the study region, sulfate and organic aerosol mass were highest near the surface and decreased rapidly with increasing altitude. The relative fraction of organic mass to sulfate was similar throughout all altitudes within the boundary layer (altitude less than 2.5 km), but was significantly higher at altitude layers in the free troposphere (above 2.5 km). A number of distinct biomass burning plumes from fires in Alaska and the Yukon were periodically intercepted, mostly at altitudes between 3 and 4 km. These plumes were associated with highest aerosol concentrations of the study and were largely comprised of organic aerosol components (approximtely 60%).

Peltier, R. E.↗

Enhanced Microbial Detection Capabilities by a Rapid Portable Instrument

We present data describing a progression of continuing technology development - from expanding the detection capabilities of the current PTS unit to re-outfitting the instrument with a protein microarray increasing the number of detectable compounds. To illustrate the adaptability of the cartridge format, on-orbit operations data from the ISS demonstrate the detection of the fungal cell wall compound beta-glucan using applicable LOCAD-PTS cartridges. LOCAD-PTS is a handheld device consisting of a spectrophotometer, an onboard pumping mechanism, and data storage capabilities. A suite of interchangeable cartridges lined with four distinct capillaries allow a hydrated sample to mix with necessary reagents in the channels before being pumped to the optical well for spectrophotometric analysis. The reagents housed in one type of cartridge trigger a reaction based on the Limulus Amebocyte Lysate (LAL) assay, which results in the release of paranitroaniline dye. The dye is measured using a 395 nm filter. The LAL assay detects the Gram-negative bacterial cell wall molecule, endotoxin or lipopolysaccharide (LPS). The more dye released, the greater the concentration of endotoxin in the sample. Sampling, quantitative analysis, and data retrieval require less than 20 minutes. This is significantly faster than standard culture-based methods, which require at least a 24 hour incubation period.Using modified cartridges, we demonstrate the detection of Gram negative bacteria with protein microarray technology. Additionally, we provide data from multiple field tests where both standard and advanced PTS technologies were used. These tests investigate the transfer of target microbial molecules from one surface to another. Collectively, these data demonstrate that the new cartridges expand the number of compounds detected by LOCAD-PTS, while maintaining the rapid, in situ analysis characteristic of the instrument. The unit provides relevant data for verifying sterile sample collection protocols, which are critical for conducting accurate scientific experiments during future missions to the Moon and Mars.

Morris, Heather↗

Spectral Absorption Properties of Aerosol Particles from 350-2500nm

The aerosol spectral absorption efficiency (alpha (sub a) in square meters per gram) is measured over an extended wavelength range (350 2500 nm) using an improved calibrated and validated reflectance technique and applied to urban aerosol samples from Sao Paulo, Brazil and from a site in Virginia, Eastern US, that experiences transported urban/industrial aerosol. The average alpha (sub a) values (approximately 3 square meters per gram at 550 nm) for Sao Paulo samples are 10 times larger than alpha (sub a) values obtained for aerosols in Virginia. Sao Paulo aerosols also show evidence of enhanced UV absorption in selected samples, probably associated with organic aerosol components. This extra UV absorption can double the absorption efficiency observed from black carbon alone, therefore reducing by up to 50% the surface UV fluxes, with important implications for climate, UV photolysis rates, and remote sensing from space.

Martins, J. Vanderlei↗

Low Fluid Shear Culture of Staphylococcus Aureus Represses hfq Expression and Induces an Attachment-Independent Biofilm Phenotype

Background: The opportunistic pathogen, Staphylococcus aureus, experiences fluctuations in fluid shear during infection and colonization of a human host. Colonization frequently occurs at mucus membrane sites such as in the gastrointestinal tract where the bacterium may experience low levels of fluid shear. The response of S. aureus to low fluid shear remains unclear. Methods: S. aureus was cultured to stationary phase using Rotating-Wall Vessel (RWV) bioreactors which produce a physiologically relevant low fluid shear environment. The bacterial aggregates that developed in the RWV were evaluated by electron microscopy as well as for antibiotic resistance and other virulence-associated stressors. Genetic expression profiles for the low-shear cultured S. aureus were determined by microarray analysis and quantitative real-time PCR. Results: Planktonic S. aureus cultures in the low-shear environment formed aggregates completely encased in high amounts of extracellular polymeric substances. In addition, these aggregates demonstrated increased antibiotic resistance indicating attachment-independent biofilm formation. Carotenoid production in the low-shear cultured S. aureus was significantly decreased, and these cultures displayed an increased susceptibility to oxidative stress and killing by whole blood. The hfq gene, associated with low-shear growth in Gram negative organisms, was also found to be down-regulated in S. aureus. Conclusions: Collectively, this data suggests that S. aureus decreases virulence characteristics in favor of a biofilm-dwelling colonization phenotype in response to a low fluid shear environment. Furthermore, the identification of an Hfq response to low-shear culture in S. aureus, in addition to the previously reported responses in Gram negative organisms, strongly suggests an evolutionarily conserved response to mechanical stimuli among structurally diverse prokaryotes.

Ott, C. Mark↗