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Fundamental Study of Material Flow in Friction Stir Welds

The presented research project consists of two major parts. First, the material flow in solid-state, friction stir, butt-welds as been investigated using a marker insert technique. Changes in material flow due to welding parameter as well as tool geometry variations have been examined for different materials. The method provides a semi-quantitative, three-dimensional view of the material transport in the welded zone. Second, a FSW process model has been developed. The fully coupled model is based on fluid mechanics; the solid-state material transport during welding is treated as a laminar, viscous flow of a non-Newtonian fluid past a rotating circular cylinder. The heat necessary for the material softening is generated by deformation of the material. As a first step, a two-dimensional model, which contains only the pin of the FSW tool, has been created to test the suitability of the modeling approach and to perform parametric studies of the boundary conditions. The material flow visualization experiments agree very well with the predicted flow field. Accordingly, material within the pin diameter is transported only in the rotation direction around the pin. Due to the simplifying assumptions inherent in the 2-D model, other experimental data such as forces on the pin, torque, and weld energy cannot be directly used for validation. However, the 2-D model predicts the same trends as shown in the experiments. The model also predicts a deviation from the "normal" material flow at certain combinations of welding parameters, suggesting a possible mechanism for the occurrence of some typical FSW defects. The next step has been the development of a three-dimensional process model. The simplified FSW tool has been designed as a flat shoulder rotating on the top of the workpiece and a rotating, cylindrical pin, which extends throughout the total height of the flow domain. The thermal boundary conditions at the tool and at the contact area to the backing plate have been varied to fit experimental data such as temperature profiles, torque and tool forces. General aspects of the experimentally visualized material flow pattern are confirmed by the 3-D model.

Reynolds, Anthony P.↗

Melt Redistribution in Dynamic Systems: Applications to Core Formation

Samples of olivine + Fe-S were fabricated by hot-pressing a mechanical mixture of the two phases. Olivine powders with a starting grain size of approx. 10 microns were prepared by crushing crystals of San Carlos olivine followed by pulverization in a fluid energy mill. Iron sulfide powder with a particle size of approx. 5 microns was prepared from a 2:1 mixture of reagent grade iron sulfide (FeS) plus iron. Rods 20 mm in length by 10 mm in diameter were cold-pressed into iron capsules with a uniaxial stress of approx. 200 MPa. These rods were then hot-pressed at 1523 K and 300 MPa for 4 h to obtain a starting material with <2% porosity. Samples for shear experiments were cut perpendicular to the long axis of these rods. The discs were shaped into ellipses approx. 6 mm by approx. 8 mm with a thickness ranging from 0.6 to 1.0 mm. An iron foil strain marker was inserted into a cut made along the minor axis. The sample was placed between two thoriated tungsten pistons cut at 45 deg to the long axis and then placed into an iron sleeve capped by alumina discs. The sample was then sheared in a gas pressure-medium apparatus at a temperature of 1523 K and a confining pressure of 300 MPa.

Hustoft, J. W.↗

DNA-directed mutations. Leading and lagging strand specificity

The fidelity of replication has evolved to reproduce B-form DNA accurately, while allowing a low frequency of mutation. The fidelity of replication can be compromised, however, by defined order sequence DNA (dosDNA) that can adopt unusual or non B-DNA conformations. These alternative DNA conformations, including hairpins, cruciforms, triplex DNAs, and slipped-strand structures, may affect enzyme-template interactions that potentially lead to mutations. To analyze the effect of dosDNA elements on spontaneous mutagenesis, various mutational inserts containing inverted repeats or direct repeats were cloned in a plasmid containing a unidirectional origin of replication and a selectable marker for the mutation. This system allows for analysis of mutational events that are specific for the leading or lagging strands during DNA replication in Escherichia coli. Deletions between direct repeats, involving misalignment stabilized by DNA secondary structure, occurred preferentially on the lagging strand. Intermolecular strand switch events, correcting quasipalindromes to perfect inverted repeats, occurred preferentially during replication of the leading strand.

NASA Discipline Radiation Health↗

A novel root gravitropism mutant of Arabidopsis thaliana exhibiting altered auxin physiology

A root gravitropism mutant was isolated from the DuPont Arabidopsis thaliana T-DNA insertional mutagenesis collection. This mutant has reduced root gravitropism, hence the name rgr1. Roots of rgr1 are shorter than those of wild-type, and they have reduced lateral root formation. In addition, roots of rgr1 coil clockwise on inclined agar plates, unlike wild-type roots which grow in a wavy pattern. The rgr1 mutant has increased resistance, as measured by root elongation, to exogenously applied auxins (6-fold to indole-3-acetic acid, 3-fold to 2,4-dichlorophenoxyacetic acid, and 2-fold to napthyleneacetic acid). It is also resistant to polar auxin transport inhibitors (2-fold to triiodobenzoic acid and 3- to 5-fold to napthylphthalamic acid). The rgr1 mutant does not appear to be resistant to other plant hormone classes. When grown in the presence of 10(-7) M 2,4-dichlorophenoxyacetic acid, rgr1 roots have fewer root hairs than wild type. All these rgr1 phenotypes are Mendelian recessives. Complementation tests indicate that rgr1 is not allelic to previously characterized agravitropic or auxin-resistant mutants. The rgr1 locus was mapped using visible markers to 1.4 +/- 0.6 map units from the CH1 locus at 1-65.4. The rgr1 mutation and the T-DNA cosegregate, suggesting that rgr1 was caused by insertional gene inactivation.

NASA Discipline Plant Biology↗

Determination of muscle-specific glucose flux using radioactive stereoisomers and microdialysis

The purpose of the present study was to evaluate a novel approach for determining skeletal muscle-specific glucose flux using radioactive stereoisomers and the microdialysis technique. Microdialysis probes were inserted into the vastus lateralis muscle of human subjects and perfused (4 microl/min) with a Ringer solution containing small amounts of radioactive D- and L-glucose as the internal reference markers for determining probe recovery as well as varying concentrations of insulin (0-10 microM). The rationale behind this approach was that both stereoisomers would be equally affected by the factors that determine probe recovery, with the exception of L-glucose, which is nonmetabolizable and would not be influenced by tissue uptake. Therefore, any differences in the probe recovery ratios between the D- and L-stereoisomers represent changes in skeletal muscle glucose uptake directly at the tissue level. There were no differences in probe recovery between the D- (42.3 +/- 3.5%) and L- (41.2 +/- 3.5) stereoisomers during the control period (no insulin), which resulted in a D/L ratio of 1.04 +/- 0.03. However, during insulin perfusion (1 microM), The D/L ratio increased to 1.62 +/- 0.08 and 1.58 +/- 0.07 (P < 0.05) during the two collection (0-15 and 15-30 min) periods, respectively. This was accomplished solely by an increase (P < 0.05) in D-glucose probe recovery, as L-glucose probe recovery remained unchanged. In a second set of experiments, the perfusion of 10 microM insulin did not increase the D/L ratio (1.40 +/- 0.11) above that observed during 1.0 microM (1.41 +/- 0.07) insulin perfusion. These data suggest that this method is sufficiently sensitive to detect differences in insulin-stimulated glucose uptake; thus the use of radioactive stereoisomers in conjunction with the microdialysis technique provides a novel and useful technique for determining tissue-specific glucose flux and insulin sensitivity.

Non-NASA Center↗

Computed Tomography 3-D Imaging of the Metal Deformation Flow Path in Friction Stir Welding

In friction stir welding (FSW), a rotating threaded pin tool is inserted into a weld seam and literally stirs the edges of the seam together. To determine optimal processing parameters for producing a defect free weld, a better understanding of the resulting metal deformation flow path is required. Marker studies are the principal method of studying the metal deformation flow path around the FSW pin tool. In our study, we have used computed tomography (CT) scans to reveal the flow pattern of a lead wire embedded in a FSW weld seam. At the welding temperature of aluminum, the lead becomes molten and is carried with the macro-flow of the weld metal. By using CT images, a 3-dimensional (3D) image of the lead flow pattern can be reconstructed. CT imaging was found to be a convenient and comprehensive way of collecting and displaying tracer data. It marks an advance over previous more tedious and ambiguous radiographic/metallographic data collection methods.

Schneider, Judy↗

Computer Tomography 3-D Imaging of the Metal Deformation Flow Path in Friction Stir Welding

In friction stir welding, a rotating threaded pin tool is inserted into a weld seam and literally stirs the edges of the seam together. This solid-state technique has been successfully used in the joining of materials that are difficult to fusion weld such as aluminum alloys. To determine optimal processing parameters for producing a defect free weld, a better understanding of the resulting metal deformation flow path is required. Marker studies are the principal method of studying the metal deformation flow path around the FSW pin tool. In our study, we have used computed tomography (CT) scans to reveal the flow pattern of a lead wire embedded in a FSW weld seam. At the welding temperature of aluminum, the lead becomes molten and thus tracks the aluminum deformation flow paths in a unique 3-dimensional manner. CT scanning is a convenient and comprehensive way of collecting and displaying tracer data. It marks an advance over previous more tedious and ambiguous radiographic/metallographic data collection methods.

Schneider, Judy↗

The Potassium-Argon Laser Experiment (KarLE): In Situ Geochronology for Mars and Beyond

The search for life in the solar system depends upon discovering the right moments in planetary evolution: when habitable environments existed, when they declined, and when geologic processes operated to preserve traces of life after death. However, an incomplete knowledge of absolute Martian geochronology limits our ability to understand the timing of Martian evolutionary milestones, major climate changes, and stratigraphic epochs [1, 2]. Absolute dating relates these habitability markers to planetarywide geologic, atmospheric, and climate history places, and ties their occurrence to the history of the solar system, especially the Earth-Moon system and the timescale of evolution of life on Earth. KArLE is being developed to anchor the relative timeline of geological events to an absolute chronology that puts Mars into a wider solar system context. KArLE makes its measurements on rock samples that can be obtained by landers or rovers and inserted into a small, mechanically simple chamber. KArLE interrogates the samples using laser-induced breakdown spectrocopy (LIBS), mass spectrometry, and optical imaging. The KArLE experiment is flexible enough to accommodate any partner providing these instrument components, a creative approach that extends the ability of mission payloads to accomplish an additional highly-desirable science measurement for low cost and risk and minimal extra hardware.

Cohen, Barbara A.↗

Flow in the Proximity of the Pin-Tool in Friction Stir Welding and Its Relation to Weld Homogeneity

In the Friction Stir Welding (FSW) process a rotating pin inserted into a seam literally stirs the metal from each side of the seam together. It is proposed that the flow in the vicinity of the pin-tool comprises a primary rapid shear over a cylindrical envelope covering the pin-tool and a relatively slow secondary flow taking the form of a ring vortex about the tool circumference. This model is consistent with a plastic characterization of metal flow, where discontinuities in shear flow are allowed but not viscous effects. It is consistent with experiments employing several different kinds of tracer: atomic markers, shot, and wire. If a rotating disc with angular velocity w is superposed on a translating continuum with linear velocity omega, the trajectories of tracer points become circular arcs centered upon a point displaced laterally a distance v/omega from the center of rotation of the disc in the direction of the advancing side of the disc. In the present model a stream of metal approaching the tool (taken as the coordinate system of observation) is sheared at the slip surface, rapidly rotated around the tool, sheared again on the opposite side of the tool, and deposited in the wake of the tool. Local shearing rates are high, comparable to metal cutting in this model. The flow patterns in the vicinity of the pin-tool determine the level of homogenization and dispersal of contaminants that occurs in the FSW process. The approaching metal streams enfold one another as they are rotated around the tool. Neglecting mixing they return to the same lateral position in the wake of the tool preserving lateral tracer positions as if the metal had flowed past the tool like an extrusion instead of being rotated around it. (The seam is, however, obliterated.) The metal stream of thickness approximately that of the tool diameter D is wiped past the tool at elevated temperatures drawn out to a thickness of v/2(omega) in the wiping zone. Mixing distances in the wiping zone are multiplied in the unfolded metal. Inhomogeneities on a smaller scale than the mixing length are obliterated, but structure on a larger scale may be transmitted to the wake of a FSW weld.

Nunes, Arthur C., Jr.↗

Microelectronic DNA assay for the detection of BRCA1 gene mutations

Mutations in BRCA1 are characterized by predisposition to breast cancer, ovarian cancer and prostate cancer as well as colon cancer. Prognosis for this cancer survival depends upon the stage at which cancer is diagnosed. Reliable and rapid mutation detection is crucial for the early diagnosis and treatment. We developed an electronic assay for the detection of a representative single nucleotide polymorphism (SNP), deletion and insertion in BRCA1 gene by the microelectronics microarray instrumentation. The assay is rapid, and it takes 30 minutes for the immobilization of target DNA samples, hybridization, washing and readout. The assay is multiplexing since it is carried out at the same temperature and buffer conditions for each step. The assay is also highly specific, as the signal-to-noise ratio is much larger than recommended value (72.86 to 321.05 vs. 5) for homozygotes genotyping, and signal ratio close to the perfect value 1 for heterozygotes genotyping (1.04).

Validation Studies↗

Flow Trajectories in Friction Stir Welding

In the Friction Stir Welding (FSW) process, a rotating, shouldered tool with a threaded pin is inserted under pressure along the seam of two pieces of metal which are tightly clamped together, and secured against a rigid anvil underneath. The rotating pin travels along the seam and through a combination of pressure and friction heating produces a zone of plastic deformation around the pin within the workpiece on either side of the seam. As the pin is moved in the direction of welding, the plasticised material moves around the tool and bonds together behind it. The elements of the material flow behavior are a combination of three elements. There is a rotational transport of material being carried around the tool, extrusion of material being forced around the pin on both sides into the cavity created behind it, and a lifting and dropping of material as it is stirred and mixed by the rotating action of the pin. It was assumed that rotational motion of the plastic zone is the primary mechanism for transport of material around the welding tool. A kinematic mathematical model was used to compute trajectories of material movement for various distributions of rotational slip within the plastic zone. These trajectories were then compared with the results of an experiment that produced radiographs of markers embedded in a workpiece that was welded with the FSW process. It was assumed that the copper wire markers retained their original length as the aluminum material flowed around them. The kinematic model included a constraint so that the displacements were such that the total length of the wire markers did not increase after deformation. There was good agreement between the calculated trajectories for the case of localized slip at the outer surface of the plastic deformation zone and the radiographs of the copper wire markers. The trajectories differed markedly from the radiograph traces when a distributed slip zone was assumed. It was concluded that the flow field could be described generally by a rotating plug model with slip occurring on the surface at the outer boundary of the plastic zone.

Bernstein, Edward L.↗

Simulated Space Radiation: Murine Skeletal Responses During Recovery and with Mechanical Stimulation

Simulated space radiation at doses similar to those of solar particle events or a round-trip sojourn to Mars (1-2Gy) may cause skeletal tissue degradation and deplete stem/progenitor cell pools throughout the body. We hypothesized that simulated space radiation (SSR) causes late, time-dependent deficits in bone structure and bone cell function reflected by changes in gene expression in response to anabolic stimuli. We used a unique sequential dual ion exposure (proton and iron) for SSR to investigate time-dependence of responses in gene expression, cell function, and microarchitecture with respect to radiation and an anabolic stimulus of axial loading (AL). Male 16-wk C57BL6/J mice (n=120 total) were exposed to 0Gy (Sham, n=10), 56Fe (2Gy, positive control dose, n=10), or sequential ions for SSR (1Gy 1H/56Fe/1H, n=10) by total body irradiation (IR), and the tissues were harvested 2 or 6 mo. later. Further, to assess the response to anabolic stimuli, we subjected additional Sham-AL (n=15) and SSR-AL (n=15) groups to rest-inserted tibial axial loading (AL) starting at 1 and 5 months post-IR (-9N, 60 cycles/day, 3 days/wk, 4 wks). Exposure to 56Fe caused a significant reduction in cancellous bone volume fraction (BV/TV) compared to Sham (-34%) and SSR (-20%) in the proximal tibia metaphysis at 2-months post-IR; however BV/TV for SSR group was not different than Sham. Both 56Fe and SSR caused significant reduction in trabecular number (Tb.N) compared to Sham (-33% and -16%, respectively). Further, Tb.N for 56Fe (2Gy) was significantly lower than SSR (-21%). Ex vivo culture of marrow cells to assess growth and differentiation of osteoblast lineage cells 6 months post-IR showed that both 56Fe and SSR exposures significantly impaired colony formation compared to Sham (-66% and -54%, respectively), as well as nodule mineralization (-90% and -51%, respectively). Two-way analysis of variance showed that both mechanical loading and radiation reduced BV/TV, mechanical loading reduced trabecular thickness (Tb.Th), and radiation reduced Tb.N, at both time points. To assess acute response to mechanical stimuli, samples were harvested from a subset of Sham-AL (n=5) and SSR-AL (n=5) to measure changes in gene expression levels. Preliminary results indicate that axial loading increased expression of the antioxidant response gene Nfe2l2 and the osteoprogenitor-associated marker Runx2 in the bone marrow cells, and there was an interaction effect between axial loading and radiation at 2-months post-IR. Additional analyses of gene expression levels in the mineralized tissue are in progress. Results indicate that SSR caused persistent impairment of osteoblast colony formation and nodule mineralization 6-mo post-IR. Contrary to our hypothesis, simulated space radiation did not impair the ability of cancellous bone to respond to a mechanical anabolic stimulus, consistent with our previous findings [1]. Hence, compressive loading may be a potential countermeasure against spaceflight-induced bone loss.

Bone Loss↗

Simulated Space Radiation: Murine Skeletal Responses During Recovery and with Mechanical Stimulation

Simulated space radiation at doses similar to those of solar particle events or a round-trip sojourn to Mars (1-2Gy) may cause skeletal tissue degradation and deplete stem/progenitor cell pools throughout the body. We hypothesized that simulated space radiation (SSR) causes late, time-dependent deficits in bone structure and bone cell function reflected by changes in gene expression in response to anabolic stimuli. We used a unique sequential dual ion exposure (proton and iron) for SSR to investigate time-dependence of responses in gene expression, cell function, and microarchitecture with respect to radiation and an anabolic stimulus of axial loading (AL). Male 16-wk C57BL6/J mice (n=120 total) were exposed to 0Gy (Sham, n=10), 56Fe (2Gy, positive control dose, n=10), or sequential ions for SSR (1Gy 1H/56Fe/1H, n=10) by total body irradiation (IR), and the tissues were harvested 2 or 6 mo. later. Further, to assess the response to anabolic stimuli, we subjected additional Sham-AL (n=15) and SSR-AL (n=15) groups to rest-inserted tibial axial loading (AL) starting at 1 and 5 months post-IR (-9N, 60 cycles/day, 3 days/wk, 4 wks). Exposure to 56Fe caused a significant reduction in cancellous bone volume fraction (BV/TV) compared to Sham (-34%) and SSR (-20%) in the proximal tibia metaphysis at 2-months post-IR; however BV/TV for SSR group was not different than Sham. Both 56Fe and SSR caused significant reduction in trabecular number (Tb.N) compared to Sham (-33% and -16%, respectively). Further, Tb.N for 56Fe (2Gy) was significantly lower than SSR (-21%). Ex vivo culture of marrow cells to assess growth and differentiation of osteoblast lineage cells 6 months post-IR showed that both 56Fe and SSR exposures significantly impaired colony formation compared to Sham (-66% and -54%, respectively), as well as nodule mineralization (-90% and -51%, respectively). Two-way analysis of variance showed that both mechanical loading and radiation reduced BV/TV, mechanical loading reduced trabecular thickness (Tb.Th), and radiation reduced Tb.N, at both time points. To assess acute response to mechanical stimuli, samples were harvested from a subset of Sham-AL (n=5) and SSR-AL (n=5) to measure changes in gene expression levels. Preliminary results indicate that axial loading increased expression of the antioxidant response gene Nfe2l2 and the osteoprogenitor-associated marker Runx2 in the bone marrow cells, and there was an interaction effect between axial loading and radiation at 2-months post-IR. Additional analyses of gene expression levels in the mineralized tissue are in progress. Results indicate that SSR caused persistent impairment of osteoblast colony formation and nodule mineralization 6-mo post-IR. Contrary to our hypothesis, simulated space radiation did not impair the ability of cancellous bone to respond to a mechanical anabolic stimulus, consistent with our previous findings. Hence, compressive loading may be a potential countermeasure against spaceflight-induced bone loss.

MURINE SKELETAL↗

A low molecular weight artificial RNA of unique size with multiple probe target regions

Artificial RNAs (aRNAs) containing novel sequence segments embedded in a deletion mutant of Vibrio proteolyticus 5S rRNA have previously been shown to be expressed from a plasmid borne growth rate regulated promoter in E. coli. These aRNAs accumulate to high levels and their detection is a promising tool for studies in molecular microbial ecology and in environmental monitoring. Herein a new construct is described which illustrates the versatility of detection that is possible with aRNAs. This 3xPen aRNA construct carries a 72 nucleotide insert with three copies of a unique 17 base probe target sequence. This aRNA is 160 nucleotides in length and again accumulates to high levels in the E. coli cytoplasm without incorporating into ribosomes. The 3xPen aRNA illustrates two improvements in detection. First, by appropriate selection of insert size, we obtained an aRNA which provides a unique and hence, easily quantifiable peak, on a high resolution gel profile of low molecular weight RNAs. Second, the existence of multiple probe targets results in a nearly commensurate increase in signal when detection is by hybridization. These aRNAs are naturally amplified and carry sequence segments that are not found in known rRNA sequences. It thus may be possible to detect them directly. An experimental step involving RT-PCR or PCR amplification of the gene could therefore be avoided.

Non-NASA Center↗

Mechanotransductive Regulation of Gap-Junction Activity Between MLO-Y4 Osteocyte-Like and MC3T3-E1 Osteoblast-Like Cells in Three-Dimensional Co-Culture.

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.

mechanical loading↗

Mechanotransductive Regulation of Gap-Junction Activity Between MLO-Y4 Osteocyte-Like and MC3T3-E1 Osteoblast-Like Cells in Three-Dimensional Co-Culture

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.

Juran, C. M.↗

Acoustic Phased Array Quantification of Quiet Technology Demonstrator 3 Advanced Inlet Liner Noise Component

Acoustic phased array flyover noise measurements were acquired as part of the Boeing 737 MAX-7 NASA Advanced Inlet Liner segment of the Quiet Technology Demonstrator 3 (QTD3) flight test program. This paper reports on the processes used for separating and quantifying the engine inlet, exhaust and airframe noise source components and provides sample phased array-based comparisons of the component noise source levels associated with the inlet liner treatment configurations. Full scale flyover noise testing of NASA advanced inlet liners was conducted as part of the Quiet Technology Demonstrator 3 flight test program in July and August of 2018. Details on the inlet designs and testing are provided in the companion paper of Reference 1. The present paper provides supplemental details relating to the acoustic phased array portion of the analyses provided in Ref. 1. In brief, the test article was a Boeing 737MAX-7 aircraft with a modified right hand (starboard side) engine inlet, which consisted of either a production inlet liner, a NASA designed inlet liner or a simulated hard wall configuration (accomplished by applying speed tape over the inlet acoustic treatment areas). In all three configurations, the engine forward fan case acoustic panel was replaced with a unperforated (hardwall) panel. No other modifications to any other acoustic treatment areas were made. The left hand (port side) engine was a production engine and was flown at idle thrust for all measurements in order to isolate the effects of the inlet liners to the right hand engine. As described in Ref. 1, the NASA inlet treatment consists of laterally cut slots (cut perpendicular to the flow direction) which are designed to reduce excrescence drag while maintaining or exceeding the liner acoustic noise reduction capabilities. The NASA inlet liner consists of a Multi-Degree of Freedom (MDOF) design with two breathable septum layers inserted into each honeycomb cell [1]. The aircraft noise measurements were acquired for both takeoff (flaps 1 setting, gear up) and approach (flaps 30 gear up and gear down) configurations. The inlet and flight test configurations are summarized in Table 1. Table 1: Inlet Treatment and Flight Configurations Inlet Forward Fan Case Aircraft Production Hardwall Flaps 1, gear up; flaps 30 gear up; flaps 30 gear down NASA Hardwall Flaps 1, gear up; flaps 30 gear up; flaps 30 gear down Hardwall Hardwall Flaps 1, gear up; flaps 30 gear up; flaps 30 gear down III.Test Description and Hardware The flight testing was conducted at the Grant County airport in Moses Lake, WA, between 27 July and 6 August 2018. The noise measurement instrumentation included 8 flush dish microphones arranged in a noise certification configuration as well as an 840 microphone phased array. The flush dish microphones were used to quantify the levels and differences in levels between the various inlet treatments. The phased array was used to separate and quantify the narrowband (tonal) and broadband noise component levels from the engine inlet/exhaust and from the airframe. Phased array extraction of the broadband component was critical to this study because it allowed for the separation of the inlet component from the total airplane level noise even when it was significantly below the total level. Figure 1 provides an overview of the phased array microphone layout as well as a detailed image of an individual phased array microphone mounted in a plate holder (the microphone sensor is the dot in the center of the plate). The ground plane ensemble array microphones (referred to as “ensemble array” in this paper) were mounted in plates with “flower petal” edges designed to minimize edge scattering effects. Fig. 1 Flyover test microphone layout. The phased array configuration was the result of a progressive development of concepts originally implemented in Ref. 2 and refined over the following years, consisting namely of multiple multi-arm logarithmic spiral subarrays designed to cover overlapping frequency ranges and optimized for various aircraft emission angles. For the present case, the signals from all 840 microphones were acquired on a single system. The 840 microphones were parsed into 11 primary subarray sets spanning from smallest to largest aperture size and labeled accordingly as “a, b, …, k”, where “a” corresponds to the smallest fielded subarray and “k” corresponds to the largest aperture subarray. The apertures ranged from approximately 10 ft to 427 ft in size (in the flight direction) with the subarrays consisting of between 215 and 312 microphones. Figure 2 shows three such subarrays, k, h and a. As done in Ref. 2, microphones were shared between subarrays in order to reduce total channel count. Fig. 2 Sample subarray sizes (20° from overhead – refer to Figure 3a discussion). In addition to the above, each of the 11 primary subarray sets consisted of four subarrays optimized to provide near equivalent array spatial resolution in both the flight and lateral directions within 30 degrees of overhead (i.e., airplane directly above the center of the array), namely, at angles  of 0, ±10, ±20 and ±30 degrees relative to overhead where angle  is defined as shown in Figure 3a. This allowed for optimized aircraft noise measurements from 60 to 120 degree emission angle.6 An example of this pletharray design is shown in Figure 3b for the k subarray. When the aircraft is at overhead, the microphones indicated by the blue markers are used for beamforming. When the aircraft is at angles ±10 degrees from overhead, both the blue and red colored microphones are used, and so on for the ±20 and ±30 degree aircraft locations. See Ref. 3 for extensive details on pletharray design for aeroacoustic phased array testing. 6 In the discussions that follow, emission angle values are used. These are the angles at the time sound is emitted relative to the engine axis and are calculated based on flight path angle, body aircraft body angle with respect to the relative wind direction, and engine axis angle relative to aircraft body angle.

Brusniak, Leon↗