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At least 73 records · Page 4

Phylogenetic and physiological diversity of microorganisms isolated from a deep greenland glacier ice core

We studied a sample from the GISP 2 (Greenland Ice Sheet Project) ice core to determine the diversity and survival of microorganisms trapped in the ice at least 120,000 years ago. Previously, we examined the phylogenetic relationships among 16S ribosomal DNA (rDNA) sequences in a clone library obtained by PCR amplification from genomic DNA extracted from anaerobic enrichments. Here we report the isolation of nearly 800 aerobic organisms that were grouped by morphology and amplified rDNA restriction analysis patterns to select isolates for further study. The phylogenetic analyses of 56 representative rDNA sequences showed that the isolates belonged to four major phylogenetic groups: the high-G+C gram-positives, low-G+C gram-positives, Proteobacteria, and the Cytophaga-Flavobacterium-Bacteroides group. The most abundant and diverse isolates were within the high-G+C gram-positive cluster that had not been represented in the clone library. The Jukes-Cantor evolutionary distance matrix results suggested that at least 7 isolates represent new species within characterized genera and that 49 are different strains of known species. The isolates were further categorized based on the isolation conditions, temperature range for growth, enzyme activity, antibiotic resistance, presence of plasmids, and strain-specific genomic variations. A significant observation with implications for the development of novel and more effective cultivation methods was that preliminary incubation in anaerobic and aerobic liquid prior to plating on agar media greatly increased the recovery of CFU from the ice core sample.

Bacteria/classification/genetics/isolation & purif

Different Pathogenesis of CCR5-Using Primary HIV-1 Isolates from Non-Switch and Switch Virus Patients in Human Lymphoid Tissue Ex Vivo

CCR5-utilizing HIV-1 variants (R5) typically transmit infection and dominate its early stages, whereas emergence of CXCR4-using (X4 or R5X4) HIV-1 is often associated with disease progression. However, such a switch in co-receptor usage can only be detected in approximately onehalf of HIV-infected patients (switch virus patients), and progression to immunodeficiency may also occur in patients without detectable switch in co-receptor usage (non-switch virus patients). Here, we used a system of ex vivo-infected tonsillar tissue to compare the pathogenesis of sequential primary R5 HIV-1 isolates from the switch and non-switch patients. Inoculation of ex vivo tissue with these R5 isolates resulted in viral replication and CCR5(+)CD4(+) T cell depletion. The levels of such depletion by HIV-1 isolated from non-switch virus patients were significantly higher than those by R5 HIV-1 isolates from switch virus patients. T cell depletion seemed to be controlled by viral factors and did not significantly vary between tissues from different donors. In contrast, viral replication did not correlate with the switch status of the patients; in tissues fiom different donors it varied 30-fold and seemed to be controlled by a combination of viral and tissue factors. Nevertheless, replication-level hierarchy among sequential isolates remained constant in tissues from various donors. Viral load in vivo was higher in switch virus patients compared to non-switch virus patients. The high cytopathogenicity of CCR5(+)CD4(+) T cells by R5 HIV-1 isolates from non-switch virus patients may explain the steady decline of CD4(+) T cells in the absence of CXCR4 using virus; elimination of target cells by these isolates may limit their own replication in vivo.

Iarlsson, Ingrid

High-Speed Isolation Board for Flight Hardware Testing

There is a need to provide a portable and cost-effective galvanic isolation between ground support equipment and flight hardware such that any unforeseen voltage differential between ground and power supplies is eliminated. An interface board was designed for use between the ground support equipment and the flight hardware that electrically isolates all input and output signals and faithfully reproduces them on each side of the interface. It utilizes highly integrated multi-channel isolating devices to minimize size and reduce assembly time. This single-board solution provides appropriate connector hardware and breakout of required flight signals to individual connectors as needed for various ground support equipment. The board utilizes multi-channel integrated circuits that contain transformer coupling, thereby allowing input and output signals to be isolated from one another while still providing high-fidelity reproduction of the signal up to 90 MHz. The board also takes in a single-voltage power supply input from the ground support equipment and in turn provides a transformer-derived isolated voltage supply to power the portion of the circuitry that is electrically connected to the flight hardware. Prior designs used expensive opto-isolated couplers that were required for each signal to isolate and were time-consuming to assemble. In addition, these earlier designs were bulky and required a 2U rack-mount enclosure. The new design is smaller than a piece of 8.5 ~11-in. (.22 ~28-mm) paper and can be easily hand-carried where needed. The flight hardware in question is based on a lineage of existing software-defined radios (SDRs) that utilize a common interface connector with many similar input-output signals present. There are currently four to five variations of this SDR, and more upcoming versions are planned based on the more recent design.

Yamamoto, Clifford K.

Reynolds-Averaged Turbulence Model Assessment for a Highly Back-Pressured Isolator Flowfield

The use of computational fluid dynamics in scramjet engine component development is widespread in the existing literature. Unfortunately, the quantification of model-form uncertainties is rarely addressed with anything other than sensitivity studies, requiring that the computational results be intimately tied to and calibrated against existing test data. This practice must be replaced with a formal uncertainty quantification process for computational fluid dynamics to play an expanded role in the system design, development, and flight certification process. Due to ground test facility limitations, this expanded role is believed to be a requirement by some in the test and evaluation community if scramjet engines are to be given serious consideration as a viable propulsion device. An effort has been initiated at the NASA Langley Research Center to validate several turbulence closure models used for Reynolds-averaged simulations of scramjet isolator flows. The turbulence models considered were the Menter BSL, Menter SST, Wilcox 1998, Wilcox 2006, and the Gatski-Speziale explicit algebraic Reynolds stress models. The simulations were carried out using the VULCAN computational fluid dynamics package developed at the NASA Langley Research Center. A procedure to quantify the numerical errors was developed to account for discretization errors in the validation process. This procedure utilized the grid convergence index defined by Roache as a bounding estimate for the numerical error. The validation data was collected from a mechanically back-pressured constant area (1 2 inch) isolator model with an isolator entrance Mach number of 2.5. As expected, the model-form uncertainty was substantial for the shock-dominated, massively separated flowfield within the isolator as evidenced by a 6 duct height variation in shock train length depending on the turbulence model employed. Generally speaking, the turbulence models that did not include an explicit stress limiter more closely matched the measured surface pressures. This observation is somewhat surprising, given that stress-limiting models have generally been developed to better predict shock-separated flows. All of the models considered also failed to properly predict the shape and extent of the separated flow region caused by the shock boundary layer interactions. However, the best performing models were able to predict the isolator shock train length (an important metric for isolator operability margin) to within 1 isolator duct height.

Baurle, Robert A.

Low Temperature Characterization of Mechanical Isolators for Cryocoolers

For spacecraft applications requiring active refrigeration, exported vibrations from a cryocooler can be a concern. One approach to minimizing the vibrations transmitted from cryocoolers to the spacecraft is to mount them on mechanical isolators. Many commercial off-theshelf (COTS) mechanical isolators exist and have been characterized at room temperature for ground applications. However, the space environment presents challenges that complicate the selection process. Mechanical isolators in space must be made of materials that can withstand harsh radiation environments. Future instruments, such as the Mapping Imaging Spectrometer for Europa (MISE) on the Europa Clipper mission, are considering operating cryocoolers with low heat rejection temperatures and mounting them on mechanical isolators. However, the performance of many simple, traditional mechanical isolators at low temperatures is unknown. This paper describes the testing and results of various mechanical isolators able to withstand harsh radiation between 200 K and 300 K. The transmissibility of wire rope type isolators showed very little temperature dependence up to ~500 Hz. On the other hand, the transmissibility of silicone gel was strongly dependent on temperature as the material had an abrupt change in stiffness near its solid-solid phase change. Custom titanium flexures showed the most promise with a maximum transmissibility of 0.04 above 500 Hz and no expected temperature dependence.

Rodriguez, J.I.

Vibration mitigation using an inerter-based isolation system

Extreme dynamic environments can result in the failure of structures and equipment. To mitigate these effects, isolation systems can be used. In recent years, inerters have been studied as a part of different structural control methods. The inerter is attractive for many structural control methods because it can produce large mass effects despite having a small physical mass through the transformation of translational motion to the rotational motion of a flywheel. While inerters have been considered in structural control, experimental studies of isolation systems featuring inerters has been limited. This project seeks to study the ability of the addition of an inerter to improve the dynamic performance of isolation systems when considering a random vibration environment. The results of this study demonstrate that the inerter can have a large impact on the dynamic properties of an isolated system, including shifting the isolation mode frequency lower while reducing its amplitude. High frequency noise did result from the inclusion of the inerter but may not be of significant concern depending on the frequency range of interest. The strong performance of the inerter in a simplified isolation system observed during this work motivates future studies of inerters in isolation systems.

S.S. Abolghasemi

Effects of isolation on interferon production and hematological and immunological parameters

A 27-year-old woman was maintained in an isolated state for 131 days in Carlsbad Caverns, New Mexico. Her diet was vitamin D-depleted. Determinations on the effects of such isolation on levels and activities of peripheral blood cells that are important for hematological homeostasis and immunological function were carried out. Throughout the duration of the study, the percentage of lymphoid cells that expressed CD3, CD4, CD8, CD19, Leu 8, and other markers remained relatively constant although the absolute numbers of these cells varied. Although the percentage of natural killer (NK) cells did not vary, the activity of these cells did change. NK cell activity became elevated as the isolation study progressed. Production of interferon-gamma (IFN-gamma) in response to mitogen stimulation was higher than expected throughout the isolation periods, but returned to the normal range after termination of the isolation. Red and white cell counts dropped significantly upon entering isolation, but soon returned to normal.

NASA Discipline Cell Biology

Overexpression of Catalase in Mitochondria Mitigates Changes in Hippocampal Cytokine Expression Following Simulated Microgravity and Isolation

Isolation on Earth can alter physiology and signaling of organs systems, including the central nervous system. Although not in complete solitude, astronauts operate in an isolated environment during spaceflight. In this study, we determined the effects of isolation and simulated microgravity solely or combined, on the inflammatory cytokine milieu of the hippocampus. Adult female wild-type mice underwent simulated microgravity by hindlimb unloading for 30 days in single or social (paired) housing. In hippocampus, simulated microgravity and isolation each regulate a discrete repertoire of cytokines associated with inflammation. Their combined effects are not additive. A model for mitochondrial reactive oxygen species (ROS) quenching via targeted overexpression of the human catalase gene to the mitochondria (MCAT mice), are protected from isolation- and/or simulated microgravity-induced changes in cytokine expression. These findings suggest a key role for mitochondrial ROS signaling in neuroinflammatory responses to spaceflight and prolonged bedrest, isolation, and confinement on Earth.

simulated microgravity

Genomic analysis and identification of a novel superantigen, SargEY, in Staphylococcus argenteus isolated from atopic dermatitis lesions

During surveillance of Staphylococcus aureus in lesions from patients with atopic dermatitis (AD), we isolated Staphylococcus argenteus, a species registered in 2011 as a new member of the genus Staphylococcus and previously considered a lineage of S. aureus. Genome sequence comparisons between S. argenteus isolates and representative S. aureus clinical isolates from various origins revealed that the S. argenteus genome from AD patients closely resembles that of S. aureus causing skin infections. We previously reported that 17%–22% of S. aureus isolated from skin infections produce staphylococcal enterotoxin Y (SEY), which predominantly induces T-cell proliferation via the T-cell receptor (TCR) Vα pathway. Complete genome sequencing of S. argenteus isolates revealed a gene encoding a protein similar to superantigen SEY, designated as SargEY, on its chromosome. Population structure analysis of S. argenteus revealed that these isolates are ST2250 lineage, which was the only lineage positive for the SEY-like gene among S. argenteus. Recombinant SargEY demonstrated immunological cross-reactivity with anti-SEY serum. SargEY could induce proliferation of human CD4 + and CD8 + T cells, as well as production of TNF-α and IFN-γ. SargEY showed emetic activity in a marmoset monkey model. S arg EY and SET (a phylogenetically close but uncharacterized SE) revealed their dependency on TCR Vα in inducing human T-cell proliferation. Additionally, TCR sequencing revealed other previously undescribed Vα repertoires induced by SEH. S arg EY and SEY may play roles in exacerbating the respective toxin-producing strains in AD.

59 BASIC BIOLOGICAL SCIENCES

Survival of bacterial isolates exposed to simulated Jovian trapped radiation belt electrons and solar wind protons

With missions to Jupiter, the spacecraft will be exposed for extended duration to solar wind radiation and the Jovian trapped radiation belt. This study is designed to determine the effect of these radiation environments on spacecraft bacterial isolates. The information can be used in the probability of contamination analysis for these missions. A bacterial subpopulation from Mariner Mars 1971 spacecraft (nine sporeforming and three nonsporeforming isolates) plus two comparative organisms, Staphylococcus epidermidis ATCC 17917 and a strain of Bacillus subtilis var. niger, were exposed to 2-, 12-, and 25-MeV electrons at different doses with simultaneous exposure to a vacuum of 0.0013 N/sqm at 20 and -20 C. The radioresistance of the subpopulation was dependent on the isolate, dose, and energy of electrons. Temperature affected the radioresistance of only the sporeforming isolates. Survival data indicated that spores were reduced approximately 1 log/1500 J/kg, while nonsporeforming isolates (micrococci) were reduced 1.5 to 2 logs/1500 J/kg with the exception of an apparent radioresistant isolate whose resistance approached that of the spores. The subpopulation was found to be less resistant to lower energy than to higher energy electrons.

Taylor, D. M.

Shock isolator for diode laser operation on a closed-cycle refrigerator

Closed-cycle helium refrigerators are widely used as coolers for semiconductor diode lasers. These refrigerators pose several difficulties including temperature oscillations due to varying refrigerator capacity during the Solvay cycle, and impact shocks delivered to the diode in the cycle's expansion phase. A shock isolator has been designed to isolate diode lasers from such impact shocks. Slow diode current scans have been made before installation of the shock isolator, with the isolator but no thermal damper, and with both devices. With the isolator and no damper, the diode output frequency oscillated at the refrigerator cycle rate, deviating by plus or minus 40 MHz. Using the isolator and the damper no frequency fluctuation was detected.

Jennings, D. E.

Vibration isolation technology experiment

The objectives of the vibration isolation technology experiment are to demonstrate the viability of the magnetic suspension technology in providing the isolation of large structures elements from the external environment and to quantify the degree of isolation provided by this system. The approach proposed for this experiment is to mount a six-degrees-of-freedom magnetic bearing suspension system at the free end of a shuttle-attached flexible structure such as MAST. The disturbance generator, located on top of the isolation system, will be energized at selected and broadband frequencies to simulate a typical spacecraft vibration environment. Sensors located on the isolation system and the flexible structures element will be used to quantify the degree of isolation provided by this system.

Keckler, C. R.

NASDA's activities on vibration isolation technology

The National Space Development Agency's (NASDA) activities in providing various vibration isolation technologies for the Space Station Mission are covered in viewgraph form. Technologies covered include an active vibration isolation system for extra sensitive missions in the low frequency range, a passive damping system consisting of a damping rack for the reduction of resonance amplification, and an isolator for vibration isolation from low frequencies. Information is given in viewgraph form on the active vibration isolation concept, voice coil type electromagnetic suspension, a profile of an active vibration isolation system, a three degree of freedom ground experiment, and acceleration feedback.

Source record

Control issues of microgravity vibration isolation

Active vibration isolation systems contemplated for microgravity space experiments may be designed to reach given performance requirements in a variety of ways. An analogy to passive isolation systems proves to be illustrative but lacks the flexibility as a design tool of a control systems approach and may lead to poor designs. For example, it is shown that a focus on equivalent stiffness in isolation system design leads to a controller that sacrifices robustness for performance. Control theory as applied to vibration isolation is reviewed and passive analogies are discussed. The loop shaping trade-off is introduced and used to design a single-degree-of-freedom fedback controller. An algebraic control design methodology is contrasted to loop shaping and critiqued. Multi-axis vibration isolation and the problems of decoupled single loop control are introduced through a two-degree-of-freedom example problem. It is shown that center of mass uncertainty may result in instability when decoupled single loop control is used. This results from the ill-conditioned nature of the feedback control design. The use of the Linear Quadratic Regulator synthesis procedure for vibration isolation controller design is discussed.

Knospe, C. R.

Hardware interface for isolation of vibrations in flexible manipulators: Development and applications

NASA's Langley Research Center (LaRC) is addressing the problem of isolating the vibrations of the Shuttle remote manipulator system (RMS) from its end-effector and/or payload by modeling an RMS flat-floor simulator with a dynamic payload. Analysis of the model can lead to control techniques that will improve the speed, accuracy, and safety of the RMS in capturing satellites and eventually facilitate berthing with the space station. Rockwell International Corporation, also involved in vibration isolation, has developed a hardware interface unit to isolate the end-effector from the vibrations of an arm on a Shuttle robotic tile processing system (RTPS). To apply the RTPS isolation techniques to long-reach arms like the RMS, engineers have modeled the dynamics of the hardware interface unit with simulation software. By integrating the Rockwell interface model with the NASA LaRC RMS simulator model, investigators can study the use of a hardware interface to isolate dynamic payloads from the RMS. The interface unit uses both active and passive compliance and damping for vibration isolation. Thus equipped, the RMS could be used as a telemanipulator with control characteristics for capture and berthing operations. The hardware interface also has applications in industry.

Manouchehri, Davoud

Active thermal isolation for temperature responsive sensors

The detection of flow transition between laminar and turbulent flow and of shear stress or skin friction of airfoils is important in basic research for validation of airfoil theory and design. These values are conventionally measured using hot film nickel sensors deposited on a polyimide substrate. The substrate electrically insulates the sensor and underlying airfoil but is prevented from thermally isolating the sensor by thickness constraints necessary to avoid flow contamination. Proposed heating of the model surface is difficult to control, requires significant energy expenditures, and may alter the basic flow state of the airfoil. A temperature responsive sensor is located in the airflow over the specified surface of a body and is maintained at a constant temperature. An active thermal isolator is located between this temperature responsive sensor and the specific surface of the body. The total thickness of the isolator and sensor avoid any contamination of the flow. The temperature of this isolator is controlled to reduce conductive heat flow from the temperature responsive sensor to the body. This temperature control includes (1) operating the isolator at the same temperature as the constant temperature of the sensor; and (2) establishing a fixed boundary temperature which is either less than or equal to, or slightly greater than the sensor constant temperature. The present invention accordingly thermally isolates a temperature responsive sensor in an energy efficient, controllable manner while avoiding any contamination of the flow.

Martinson, Scott D.

Microgravity isolation system design: A case study

Many acceleration-sensitive, microgravity science experiments will require active vibration isolation from manned orbiters on which they will be mounted. The isolation problem, especially in the case of a tethered payload, is a complex three-dimensional one that is best suited to modern-control design methods. In this paper, extended H(sub 2) synthesis is used to design an active isolator (i.e., controller) for a realistic single-input-multiple-output (SIMO) microgravity vibration isolation problem. Complex mu-analysis methods are used to analyze the isolation system with respect to sensor, actuator, and umbilical uncertainties. The paper fully discusses the design process employed and the insights gained. This design case study provides a practical approach for isolation problems of greater complexity. Issues addressed include a physically intuitive state-space description of the system, disturbance and noise filters, filters for frequency weighting, and uncertainty models. The controlled system satisfies all the performance specifications and is robust with respect to model uncertainties.

Hampton, R. D.

Experimental investigation of inlet-combustor isolators for a dual-mode scramjet at a Mach number of 4

This report details experimentally derived operational characteristics of numerous two-dimensional planar inlet-combustor isolator configurations at a Mach number of 4. Variations in geometry included (1) inlet cowl length; (2) inlet cowl rotation angle; (3) isolator length; and (4) utilization of a rearward-facing isolator step. To obtain inlet-isolator maximum pressure-rise data relevant to ramjet-engine combustion operation, configurations were mechanically back pressured. Results demonstrated that the combined inlet-isolator maximum back-pressure capability increases as a function of isolator length and contraction ratio, and that the initiation of unstart is nearly independent of inlet cowl length, inlet cowl contraction ratio, and mass capture. Additionally, data are presented quantifying the initiation of inlet unstarts and the corresponding unstart pressure levels.

Emami, Saied