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At least 37 records · Page 2

Ambient fine particulate matters induce cell death and inflammatory response by influencing mitochondria function in human corneal epithelial cells

Ambient fine particulate matter (AFP) is a main risk factor for the cornea as ultraviolet light. However, the mechanism of corneal damage following exposure to AFP has been poorly understood. In this study, we first confirmed that AFP can penetrate the cornea of mice, considering that two-dimensional cell culture systems are limited in reflecting the situation in vivo. Then, we investigated the toxic mechanism using human corneal epithelial (HCET) cells. At 24 h after exposure, AFP located within the autophagosome-like vacuoles, and cell proliferation was clearly inhibited in all the tested concentration. Production of ROS and NO and secretion of pro-inflammatory cytokines were elevated in a dose-dependent manner. Additionally, conversion of LC3B from I-type to II-type and activation of caspase cascade which show autophagic- and apoptotic cell death, respectively, were observed in cells exposed to AFP. Furthermore, AFP decreased mitochondrial volume, inhibited ATP production, and altered the expression of metabolism-related genes. Taken together, we suggest that AFP induces cell death and inflammatory response by influencing mitochondrial function in HCET cells. In addition, we recommend that stringent air quality regulations are needed for eye health. - Highlights: • In this study, we studied a toxic mechanism of AFP in human corneal epithelial cells. • AFP located within autophagosome-like vacuoles at 24 h after exposure. • AFP induced apoptotic cell death with mitochondrial dysfunction and membrane damage. • AFP elevated level of ROS, NO, and pro-inflammatory cytokines. • Levels of Apoptosis/autophagy-related protein and metabolism-related gene increased.

54 ENVIRONMENTAL SCIENCES↗

On the engulfment of antifreeze proteins by ice

Antifreeze proteins (AFPs) are remarkable biomolecules that suppress ice formation at trace concentrations. To inhibit ice growth, AFPs must not only bind to ice crystals, but also resist engulfment by ice. The highest supercooling, ΔT*, for which AFPs are able to resist engulfment is widely believed to scale as the inverse of the separation, L, between bound AFPs, whereas its dependence on the molecular characteristics of the AFP remains poorly understood. By using specialized molecular simulations and interfacial thermodynamics, here, we show that in contrast with conventional wisdom, ΔT* scales as L –2 and not as L –1 . We further show that ΔT* is proportional to AFP size and that diverse naturally occurring AFPs are optimal at resisting engulfment by ice. By facilitating the development of AFP structure–function relationships, we hope that our findings will pave the way for the rational design of AFPs.

Science & Technology - Other Topics↗

Revealing Surface Waters on an Antifreeze Protein by Fusion Protein Crystallography Combined with Molecular Dynamic Simulations

Antifreeze proteins (AFPs) adsorb to ice through an extensive, flat, relatively hydrophobic surface. It has been suggested that this ice-binding site (IBS) organizes surface waters into an ice-like clathrate arrangement that matches and fuses to the quasi-liquid layer on the ice surface. On cooling, these waters join the ice lattice and freeze the AFP to its ligand. Evidence for the generality of this binding mechanism is limited because AFPs tend to crystallize with their IBS as a preferred protein–protein contact surface, which displaces some bound waters. Type III AFP is a 7 kDa globular protein with an IBS made up two adjacent surfaces. In the crystal structure of the most active isoform (QAE1), the part of the IBS that docks to the primary prism plane of ice is partially exposed to solvent and has clathrate waters present that match this plane of ice. The adjacent IBS, which matches the pyramidal plane of ice, is involved in protein–protein crystal contacts with few surface waters. Here we have changed the protein–protein contacts in the ice-binding region by crystallizing a fusion of QAE1 to maltose-binding protein. In this 1.9 Å structure, the IBS that fits the pyramidal plane of ice is exposed to solvent. By combining crystallography data with MD simulations, the surface waters on both sides of the IBS were revealed and match well with the target ice planes. Here, the waters on the pyramidal plane IBS were loosely constrained, which might explain why other isoforms of type III AFP that lack the prism plane IBS are less active than QAE1. The AFP fusion crystallization method can potentially be used to force the exposure to solvent of the IBS on other AFPs to reveal the locations of key surface waters.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Engineering Antifreeze Proteins to Optimally Resist Engulfment by Ice

Antifreeze proteins (AFPs) facilitate the survival of organisms in cold climates by inhibiting the growth and/or recrystallization of ice. To function, AFPs must first bind to ice crystals; bound AFPs must then resist engulfment by using their nonbinding side (NBS) to pin the ice–water interface. Here, we seek to understand how the molecular characteristics of an NBS, such as its ice-phobicity or shape, influence its ability to resist engulfment. By characterizing the free energy barriers that impede the engulfment of model AFPs, we find that the critical supercooling ΔT*, above which an AFP is engulfed, is dictated by an optimal pinning site on the NBS. We further find that the optimal pinning site is determined by an interplay between the contact line perimeter P and a pinning efficiency η, with ΔT* ∝ ηP at the optimal pinning site. For a hemispherical AFP, which displays progressively inward tapering, we find that η increases during engulfment, whereas P decreases; conversely, an NBS with outward tapering can achieve high P, but it suffers from low η. Because the product of η and P determines ΔT*, the inverse correlation between them limits ΔT. To circumvent such limiting behavior, we propose an NBS shape with an outward bulge; by initially tapering outward, a bulged NBS permits higher P, and by subsequently tapering inward, it promotes high η as well. Importantly, we find that ΔT* is enhanced by more than a factor of 2 with an outward bulge of only 1 nm. We also find that the more ice-phobic an NBS is, the more efficiently it pins the ice–water interface, resulting in a higher ΔT. Furthermore, our findings shed light on how the NBS molecular characteristics influence ΔT* and suggest strategies for engineering the NBS to optimally resist engulfment by ice.

Antifreeze↗

Accumulation of Antifreeze Proteins on Ice Is Determined by Adsorption

Antifreeze proteins (AFPs) enable the survival of organisms in freezing environments by binding to ice crystals and suppressing their growth. The rate at which AFPs accumulate on ice is determined by an interplay between their diffusion from the aqueous solution and their adsorption to the ice crystal. To interrogate the relative importance of these two processes, we combined fluorescence experiments with reaction–diffusion modeling. We found that as AFPs accumulate on ice, their concentration in the aqueous solution does not develop a gradient but remains equal to its bulk concentration throughout our experiments. Furthermore, these findings led us to conclude that AFP accumulation on ice crystals, which are smaller than 100 μm in radius, is not limited by the diffusion of AFPs, but by the kinetics of AFP adsorption.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Automated Fiber Placement Manufactured Composites for Science Applications

Science instruments with large collecting areas that maintain dimensional stability, such as James Webb Space Telescope and Wide Field Space Telescope, help achieve next generation science advancements. Composite materials often used for science applications include high modulus fibers in cyanate ester matrices to result in dimensionally stable structures with low contamination. Hand lay-up fabrication is the most common approach for science instrument structures. Automated Fiber Placement (AFP) using intermediate modulus fibers is commonplace in aircraft production reducing manufacturing time and increasing quality and consistency. AFP manufacturing for future large science instruments can similarly reduce costs and increase reliability. However, high modulus fibers are more prone to damage than intermediate modulus fibers. This study investigates the manufacturing viability of M55J/RS3C (Tencate) slit tape material using AFP processing. Tencate provides slit tape materials. NASA Langley Research Center (LaRC) manufactured hand layup and AFP lay-up laminates under room temperature for initial trials, Marshall Space Flight Center (MSFC) manufactured AFP laminates under room temperature and elevated temperature conditions to evaluate processing affects. Goddard Space Flight Center (GSFC) tests and evaluates tension and Coefficient of Thermal Expansion (CTE) properties by hand lay-up and AFP slit tape automated manufacturing for large science applications. These results show processing material warm reduces process induced fiber fracture; leading to stiffness and CTE properties consistent with hand lay-up, while observing a slight degradation in tensile strength.

Segal, Kenneth N.↗

Design and Analysis of a Tool for Automated Fiber Placement of Composite Wind Tunnel Blades

For decades, composite wind tunnel blades have been constructed using labor-intensive hand layup processes. Automated Fiber Placement (AFP) is a relatively new technology that has not seen widespread use in manufacturing composite wind tunnel blades. AFP offers the potential for reduced manufacturing time, reduced cost, and improved consistency compared to traditional hand layup procedures. These manufacturing qualities are becoming increasingly important as existing wind tunnel blades are replaced due to impact damage or wear and tear from decades of use. Researchers at the NASA Langley Research Center are currently using the Integrated Structural Assembly of Advanced Composites (ISAAC) facility to investigate the feasibility of AFP for manufacturing composite wind tunnel blades. ISAAC uses manufacturing tools as surfaces on which to place fiber tows. A manufacturing tool was developed for using AFP to build a wind tunnel blade shape. This paper presents the structural design and analysis of this tool, for use with AFP, constructed from high temperature thermoplastics. The tool fabricated by this design process was successfully used for AFP and oven cure of two representative airfoil shaped composite structures.

Brian Mason↗

Prognosticating Survival in Hepatocellular Carcinoma with Elevated Baseline Alpha-fetoprotein Treated with Radioembolization Using a Novel Laboratory Scoring System: Initial Development and Validation

Aims: To investigate laboratory parameters as predictors of overall survival (OS) for hepatocellular carcinoma (HCC) treated with radioembolization and develop/validate a scoring system. Methods: With IRB approval, we included all patients with baseline alpha-fetoprotein (AFP) > 100 ng/dL from our prospectively acquired HCC radioembolization database. Neutrophil–lymphocyte ratio, albumin–bilirubin (ALBI), and AFP were measured at baseline and at 1-, 3-, and 6-month post-radioembolization Landmarks. OS was assessed from these Landmarks. Univariate/multivariate analyses were performed to evaluate OS predictability of these parameters. Baseline Imaging, Laboratory, and Combination scoring systems were developed. Developing/validating groups were created to investigate/validate the score’s OS predictability. Time-dependent receiver operating characteristics (ROC) were evaluated. Patients were stratified into groups I, II, and III by using 25th and 75th percentile cutoffs according to change in Laboratory Score from baseline. Results: 345/401 (86%), 238/401 (59%), and 167/401 (42%) patients had laboratory parameters available at the 1-, 3-, and 6-month Landmarks, respectively. ALBI and AFP were significant OS prognosticators at all Landmarks. The Laboratory Score [ALBI + (0.3 × LnAFP)] was developed/internally validated to predict OS from these Landmarks. Areas under the curve of time-dependent ROCs of the Baseline Imaging vs. Laboratory scores in predicting patient OS post 3 and 6 months Landmarks were 0.56 versus 0.82 and 0.57 versus 0.77, respectively. OS differences in groups I, II, and III according to change in Laboratory Score from baseline were significant (p < 0.001). Conclusions: Post-radioembolization AFP and ALBI scores were significant OS prognosticators. A decrease in post-therapeutic Laboratory Score, which combines AFP and ALBI, correlates with an improved OS.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

Yttrium-90-Labeled Anti-Glypican 3 Radioimmunotherapy Halts Tumor Growth in an Orthotopic Xenograft Model of Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) is the second most lethal malignancy globally and is increasing in incidence in the United States. Unfortunately, there are few effective systemic treatment options, particularly for disseminated disease. Glypican-3 (GPC3) is a proteoglycan cell surface receptor overexpressed in most HCCs and provides a unique target for molecular therapies. We have previously demonstrated that PET imaging using a 89 Zr-conjugated monoclonal anti-GPC3 antibody ( α GPC3) can bind to minute tumors and allow imaging with high sensitivity and specificity in an orthotopic xenograft mouse model of HCC and that serum alpha-fetoprotein (AFP) levels are highly correlated with tumor size in this model. In the present study, we conjugated 90 Y, a high-energy beta-particle-emitting radionuclide, to our α GPC3 antibody to develop a novel antibody-directed radiotherapeutic approach for HCC. Luciferase-expressing HepG2 human hepatoblastoma cells were orthotopically implanted in the livers of athymic nude mice, and tumor establishment was verified at 6 weeks after implantation by bioluminescent imaging and serum AFP concentration. Tumor burden by bioluminescence and serum AFP concentration was highly correlated in our model. Yttrium-90 was conjugated to α GPC3 using the chelating agent 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) and injected via the tail vein into the experimental mice at a dose of 200 μ Ci/mouse or 300 μ Ci/mouse. Control mice received DOTA- α GPC3 without radionuclide. At 30 days after a single dose of the radioimmunotherapy agent, mean serum AFP levels in control animals increased dramatically, while animals treated with 200 μ Ci only experienced a minor increase, indicating cessation of tumor growth, and animals treated with 300 μ Ci experienced a reduction in serum AFP concentration, indicating tumor shrinkage. Mean tumor-bearing liver weight in control animals was also significantly greater than that in animals that received either dose of 90 Y- α GPC3. These results were achieved without significant toxicity as measured by body condition scoring and body weight. The results of this preclinical pilot demonstrate that GPC3 can be used as a target for radioimmunotherapy in an orthotopic mouse model of HCC and may be a target of clinical significance, particularly for disseminated HCC.

60 APPLIED LIFE SCIENCES↗

Advances in In-Situ Inspection of Automated Fiber Placement Systems

The advent of Automated Fiber Placement (AFP) systems have aided the rapid manufacturing of composite aerospace structures. One of the challenges that AFP systems present is the uniformity of the deposited prepreg tape layers, which are prone to laps, gaps, overlaps and twists. The current detection modus operandi involves halting fabrication and performing a time consuming visual inspection of each tape layer. Typical AFP systems use a quartz lamp to heat the base layer to make the surface tacky as it deposits another tape layer. The idea was proposed to use the preheated base layer as a through transmission heat source and to inspect the newly added tape layer using a thermographic camera. As a preliminary study of this concept a laboratory proof of concept device was designed and constructed to simulate the through transmission heat source. Using the proof of concept device, we inspected an AFP-built uncured composite specimen with artificial manufacturing defects. This paper will discuss the results of this preliminary study and the implications involved with deploying a full-scale AFP inspection system.

Juarez, Peter D.↗

In Situ Thermal Inspection of Automated Fiber Placement Operations for Tow and Ply Defect Detection

The advent of Automated Fiber Placement (AFP) systems have aided the rapid manufacturing of composite aerospace structures. One of the challenges that AFP systems pose is the uniformity of the deposited prepreg tape layers, which complicates detection of laps, gaps, overlaps and twists. The current detection method used in industry involves halting fabrication and performing a time consuming, visual inspection of each tape layer. Typical AFP systems use a quartz lamp to heat the base layer to make the surface tacky as it deposits another tape layer. The innovation proposed in this paper is to use the preheated base layer as a through-transmission heat source for inspecting the newly added tape layer in situ using a thermographic camera mounted on to the AFP hardware. Such a system would not only increase manufacturing throughput by reducing inspection times, but it would also aid in process development for new structural designs or material systems by providing data on as-built parts. To this end, a small thermal camera was mounted onto an AFP robotic research platform at NASA, and thermal data was collected during typical and experimental layup operations. The data was post processed to reveal defects such as tow overlap/gap, wrinkling, and peel-up. Defects that would have been impossible to detect visually were also discovered in the data, such as poor/loss of adhesion between plies and the effects of vacuum debulking. This paper will cover the results of our experiments, and the plans for future versions of this inspection system.

Juarez, Peter D.↗

Automated Fiber Placement Defect Identity Cards: Cause, Anticipation, Existence, Significance, and Progression

Automated Fiber Placement (AFP), a major composite manufacturing process, can result in many defects during the layup process that often require manual corrective action to produce a part with acceptable quality. These defects are the main limitation of the technology and can be hard to categorize or define in many situations. This paper provides a thorough definition and classification of all AFP defects. This effort constitutes a comprehensive and extensive library relevant to AFP defects. The defects selected and defined in this work are based on understanding and experience from the manufacture and research of advanced composite structure. Proper classification of these defects required an in-depth literature review and consideration of various viewpoints ranging from designers, manufacturers, analysts, and inspection professionals. Collectively, these sources were utilized to develop the most accurate view of each of the individual defect types. The results are presented as identity cards for each defect type, intended to provide researchers and the manufacturing industry a clear understanding of the (1) cause, (2) anticipation, (3) existence, (4) significance, and (5) progression of the defined AFP defects. The link between AFP defects and process planning, layup strategies, and machining was also investigated. Categorization of all important automated fiber placement defects is presented.

Harik, Ramy↗

Hybrid AFP/Hand Layup Optimized Blades to Enhance Wind Tunnel Capability Summary of PREDICT 2018 Efforts

A project spanning four-months in 2018 was conducted at the NASA Langley Research Center (LaRC) to advance the state of the art in Automated Fiber Placement (AFP) fabrication of complex carbon fiber wind tunnel blades for NASA wind tunnels. This effort was the first step in developing a methodology for fabricating these complex parts. An AFP robotic system was used to explore whether manufacturing time and cost for production of wind tunnel blades could be reduced. Computer aided design models of the blade surfaces were created, and AFP process planning and programming was employed to study the manufacturability of the shapes. A manufacturing demonstration unit was fabricated to demonstrate a methodology to fabricate a complex shape representative of a wind tunnel blade. The present study revealed that the heater configuration on the LaRC AFP robotic system was the primary constraint that limited its ability to use AFP for wind tunnel blade manufacturing.

carbon-epoxy↗

Adiabatic fast passage spin manipulation measurements in solid polarized targets

Adiabatic fast passage (AFP) is a rapid method for reversing nuclear polarization and manipulating spin populations in polarized solid targets, avoiding the long repolarization times associated with dynamic nuclear polarization (DNP). We report AFP measurements in a 5 T, 1 K polarized-target system for irradiated 15 NH 3 , irradiated 14 ND 3 , and butanol-based materials prepared either with TEMPO doping or by irradiation. We also present a joint manipulated-lineshape analysis for spin-1 targets and demonstrate that vector and tensor polarizations can be extracted from AFP-manipulated deuteron NMR spectra even when the populations are not described by a single Boltzmann spin temperature. In conclusion, we report a reproducible polarization- and direction-dependent AFP response in a large irradiated 15 NH 3 sample. These ammonia results are presented as empirical observations under the specific sample–coil conditions of the experiment, with possible circuit-mediated mechanisms such as radiation damping or superradiant behavior discussed but not assigned as a definitive cause.

Adiabatic fast passage↗

Development of advanced manufacturing technologies for low cost hydrogen storage vessels

The U.S. Department of Energy (DOE) defined a need for low-cost gaseous hydrogen storage vessels at 700 bar to support cost goals aimed at 500,000 units per year. Existing filament winding processes produce a pressure vessel that is structurally inefficient, requiring more carbon fiber for manufacturing reasons, than would otherwise be necessary. Carbon fiber is the greatest cost driver in building a hydrogen pressure vessel. The objective of this project is to develop new methods for manufacturing Type IV pressure vessels for hydrogen storage with the purpose of lowering the overall product cost through an innovative hybrid process of optimizing composite usage by combining traditional filament winding (FW) and advanced fiber placement (AFP) techniques. A numbers of vessels were manufactured in this project. The latest vessel design passed all the critical tests on the hybrid design per European Commission (EC) 79-2009 standard except the extreme temperature cycle test. The tests passed include burst test, cycle test, accelerated stress rupture test and drop test. It was discovered the location where AFP and FW overlap for load transfer could be weakened during hydraulic cycling at 85°C. To design a vessel that passed these tests, the in-house modeling software was updated to add capability to start and stop fiber layers to simulate the AFP process. The original in-house software was developed for filament winding only. Alternative fiber was also investigated in this project, but the added mass impacted the vessel cost negatively due to the lower performance from the alternative fiber. Overall the project was a success to show the hybrid design is a viable solution to reduce fiber usage, thus driving down the cost of fuel storage vessels. Based on DOE’s baseline vessel size of 147.3L and 91kg, the 129L vessel (scaled to DOE baseline) in this project shows a 32% composite savings and 20% cost savings when comparing Vessel 15 hybrid design and the Quantum baseline all filament wound vessel. Due to project timing, there was no additional time available to fine tune the design to improve the load transfer between AFP and FW. Further design modifications will likely help pass the extreme temperature cycle test, the remaining test that is critical to the hybrid design.

08 HYDROGEN↗

Characterization of Prepreg Tack for Composite Manufacturing by Automated Fiber Placement

Automated fiber placement (AFP) has become the industry standard for large-scale production of carbon fiber reinforced plastics (CFRP) to improve rate and reduce defects associated with manual layup. Still, defects generated during AFP processes require manual, painstaking inspection by technicians and rework of the part when substantial defects are found. Prepreg (carbon fiber infused with uncured epoxy resin) tack is one of the primary factors that influences the generation of defects that arise during auto-mated fiber placement (AFP). Tack, as it relates to AFP processes and defect formation, can be understood as a combination of two stages, cohesion and decohesion. During the cohesion phase, two pieces of prepreg are brought into contact under elevated temperature and pressure. Compaction of the resin within the contact area will result in a degree of intimate contact, I, between the mating prepreg surfaces. Defect formation, as a result of decohesion between prepreg surfaces, occurs after the cohesion phase and arises due to stress from events such as fiber placement over an existing defect, on a contoured path, etc. (Figure 1). Tack strength resists the displacement of prepreg on a surface due to stresses developed during deposition.

Wohl, Christopher J↗

Assessment of Automated Fiber Placement for the Fabrication of Composite Wind Tunnel Blades

Composite wind tunnel blades are frequently fabricated by hand layup of prepreg fabrics. Though well proven, this fabrication method is laborious and expensive. The study described in this paper used the Integrated Structural Assembly of Advanced Composites (ISAAC) facility at the NASA Langley Research Center to explore whether automated fiber placement (AFP) could reduce manufacturing time and cost for production of wind tunnel blades. Two blades, taken from two NASA wind tunnels, were investigated as representative geometries. Computer-aided design models of the blade surfaces were created, and AFP process planning and programming were employed to study the manufacturability of the shapes. A placement/cure tool was manufactured for the chosen blade surface from thermoplastic material using an additive manufacturing process. The present study revealed that the AFP head geometry, primarily the heater configuration of the ISAAC system, is the primary constraint that limits the ability to manufacture the selected wind tunnel fan blades using AFP.

Grenoble, Ray↗