Bearing Tester Data Compilation Analysis, and Reporting and Bearing Math Modeling

NASA Technical Reports Server (NTRS)

1985-01-01

The magnitude and direction of fluid induced torques and forces on the 57 mm bearing cage is considered to be a contributing factor in possible cage instabilities that can produce intermittent high heating in the bearing. Analyses of the fluid forces and torques are presented. Heat generated by viscous fluid work was estimated for two flow diverter configurations and a coolant flow of 10 lbs/sec to support the thermal evaluation of the LOX Bearing Materials Tester. Results of the analysis of the LOX turbopump turbine end bearings are discussed. Coolant velocities for the no. 4 LOX turbopump turbine end bearings were estimated as a function of shaft speed and coolant flow rate. Contact angles and track width data were developed for the 57 mm bearing as functions of shaft speed, and axial and radial loads. The Advanced Dynamics of Rolling Elements (ADORE) computer program was installed on the MSFC UNIVAC 1100 and a test case successfully run. Both the text output and the plotting output were verified. The Bearing Seal and Materials Tester - Test Condition Data Base was developed. The parametric analysis of the operating characteristics of the LOX turbopump pump end bearing using the 45 mm bearing thermal model was begun.

Development of Cre-loxP technology in zebrafish to study the regulation of fish reproduction.

PubMed

Lin, Heng-Ju; Lee, Shu-Hua; Wu, Jen-Leih; Duann, Yeh-Fang; Chen, Jyh-Yih

2013-12-01

One cannot seek permission to market transgenic fish mainly because there is no field test or any basic research on technological developments for evaluating their biosafety. Infertility is a necessary adjunct to exploiting transgenic fish unless completely secure land-locked facilities are available. In this study, we report the generation of a Cre transgenic zebrafish line using a cytomegalovirus promoter. We also produced fish carrying the Bax1 and Bax2 plasmids; these genes were separated by two loxP sites under a zona pellucida C promoter or were driven by an anti-Müllerian hormone promoter. We inserted a red fluorescent protein gene between the two loxP sites. After obtaining transgenic lines with the two transgenic fish crossed with each other (Cre transgenic zebrafish x loxP transgenic zebrafish), the floxed DNA was found to be specifically eliminated from the female or male zebrafish, and apoptosis gene expressions caused ovarian and testicular growth cessation and degeneration. Overexpression of the Bax1 and Bax2 genes caused various expression levels of apoptosis-related genes. Accordingly, this transgenic zebrafish model system provides a method to produce infertile fish and may be useful for application to genetically modified fish.

Liquid oxygen turbopump technology

NASA Technical Reports Server (NTRS)

Nielson, C. E.

1981-01-01

A small, high-pressure, LOX turbopump was designed, fabricated and tested. The pump is a single-stage centrifugal type with power to the pump supplied by a single-stage partial-admission axial-impulse turbine. Design conditions included an operating speed of 7330 rad/s (70,000 rpm), pump discharge pressure of 2977 N/sqcm (4318 psia), and a pump flowrate of 16.4 Kg/s (36.21 lb/s). The turbopump contains a self-compensating axial thrust balance piston to eliminate axial thrust loads on the bearings during steady-state operation. Testing of the turbopump was achieved usng a gaseous hydrogen high-pressure flow to drive the turbine, which generally is propelled by LOX/LH2 combustion products, at 1041K (1874 R) inlet temperature and at a design pressure ratio of 1.424. Test data obtained with the turbopump are presented which include head-flow-efficiency performance, suction performance, balance piston performance and LOX seal performance. Mechanical performance of the turbopump is also discussed.

LANTR Engine Optimization for Lunar Missions

NASA Astrophysics Data System (ADS)

Bulman, M. J.; Poth, Greg; Borowski, Stan

2006-01-01

Propulsion requirements for sustainable Lunar missions are very demanding. The high Delta V for short transit times and/or reusable vehicles are best served with the High Isp of Nuclear Propulsion. High thrust is needed to reduce gravity losses during earth departure. The LOX-Augmented Nuclear Thermal Rocket (LANTR) is a concept whereby thrust from a nuclear thermal rocket can be doubled, or even quadrupled, by the injection and combustion of gaseous oxygen downstream of the throat. This has many advantages for the mission including a reduction in the size of the reactor(s) and propellant tank volume for a given payload delivered to Low Lunar Orbit. In this paper, we conduct mission studies to define the optimum basic (Unaugmented) engine thrust, Lox augmentation level and Lox loading for minimum initial mass in low earth orbit. 35% mass savings are seen for NTR powered LTVs with over twice the propellant Volume. The LANTR powered LTV has a similar mass savings with minimal volume penalties.

Mesoporous Silica Nanoparticle-Mediated Intracellular Cre Protein Delivery for Maize Genome Editing via loxP Site Excision1,2[W][OPEN

PubMed Central

Martin-Ortigosa, Susana; Peterson, David J.; Valenstein, Justin S.; Lin, Victor S.-Y.; Trewyn, Brian G.; Lyznik, L. Alexander; Wang, Kan

2014-01-01

The delivery of proteins instead of DNA into plant cells allows for a transient presence of the protein or enzyme that can be useful for biochemical analysis or genome modifications. This may be of particular interest for genome editing, because it can avoid DNA (transgene) integration into the genome and generate precisely modified “nontransgenic” plants. In this work, we explore direct protein delivery to plant cells using mesoporous silica nanoparticles (MSNs) as carriers to deliver Cre recombinase protein into maize (Zea mays) cells. Cre protein was loaded inside the pores of gold-plated MSNs, and these particles were delivered by the biolistic method to plant cells harboring loxP sites flanking a selection gene and a reporter gene. Cre protein was released inside the cell, leading to recombination of the loxP sites and elimination of both genes. Visual selection was used to select recombination events from which fertile plants were regenerated. Up to 20% of bombarded embryos produced calli with the recombined loxP sites under our experimental conditions. This direct and reproducible technology offers an alternative for DNA-free genome-editing technologies in which MSNs can be tailored to accommodate the desired enzyme and to reach the desired tissue through the biolistic method. PMID:24376280

Operationally Efficient Propulsion System Study (OEPSS) data book. Volume 4: OEPSS design concepts

NASA Technical Reports Server (NTRS)

Wong, George S.; Ziese, James M.; Farhangi, Shahram

1990-01-01

This study was initiated to identify operations problems and cost drivers for current propulsion systems and to identify technology and design approaches to increase the operational efficiency and reduce operations costs for future propulsion systems. To provide readily usable data for the Advanced Launch System (ALS) program, the results of the OEPSS study have been organized into a series of OEPSS Data Books. This volume describes three propulsion concepts that will simplify the propulsion system design and significantly reduce operational requirements. The concepts include: (1) a fully integrated, booster propulsion module concept for the ALS that avoids the complex system created by using autonomous engines with numerous artificial interfaces; (2) an LOX tank aft concept which avoids potentially dangerous geysering in long LOX propellant lines; and (3) an air augmented, rocket engine nozzle afterburning propulsion concept that will significantly reduce LOX propellant requirements, reduce vehicle size and simplify ground operations and ground support equipment and facilities.

CFD-Based Design of Turbopump Inlet Duct for Reduced Dynamic Loads

NASA Technical Reports Server (NTRS)

Rothermel, Jeffry; Dorney, Suzanne M.; Dorney, Daniel J.

2003-01-01

Numerical simulations have been completed for a variety of designs for a 90 deg elbow duct. The objective is to identify a design that minimizes the dynamic load entering a LOX turbopump located at the elbow exit. Designs simulated to date indicate that simpler duct geometries result in lower losses. Benchmark simulations have verified that the compressible flow codes used in this study are applicable to these incompressible flow simulations.

Molecular characterization of lipoxygenase genes on chromosome 4BS in Chinese bread wheat (Triticum aestivum L.).

PubMed

Zhang, Fuyan; Chen, Feng; Wu, Peipei; Zhang, Ning; Cui, Dangqun

2015-08-01

This study cloned two novel TaLox genes on chromosome of 4BS and developed a co-dominant marker, Lox-B23, in bread wheat that showed highly significant association with lipoxygenase activity. Lipoxygenase (Lox), a critical enzyme in the carotenoid biosynthetic pathway, significantly influences the color and processing quality of wheat-based products. Two novel Lox genes, designated TaLox-B2 and TaLox-B3, were cloned on chromosome 4BS of Chinese bread wheat. The deduced amino acid sequence showed that both TaLox-B2 and TaLox-B3 genes encoded an 861-aa protein and possessed a lipoxygenase superfamily domain at the 170-838 interval. Two different TaLox-B2 alleles, designated TaLox-B2a and TaLox-B2b, were subsequently discovered. A co-dominant marker, Lox-B23, was developed based on sequences of TaLox-B2a, TaLox-B2b, and TaLox-B3 genes to precisely distinguish these three alleles in Chinese bread cultivars. Among five allelic combinations of Lox genes at Lox-B1, Lox-B2, and Lox-B3 loci, wheat cultivars with TaLox-B1a/TaLox-B2a/TaLox-B3a combination exhibited the highest Lox activity, whereas those with TaLox-B1a/TaLox-B2b/TaLox-B3b combination significantly showed the lowest Lox activity. A RIL population was used to evaluate the influence of TaLox-B3a gene on Lox activity. Results showed that TaLox-B3a gene could significantly increase the Lox activity in bread wheat. Physical mapping indicated that both TaLox-B2 and TaLox-B3 genes were located on chromosome 4BS in bread wheat. This study provides useful information to further understand the molecular and genetic bases of Lox activity in bread wheat.

Discovery LOX

NASA Image and Video Library

2003-11-11

In the Orbiter Processing Facility, workers prepare to install the liquid oxygen feedline for the 17-inch disconnect on orbiter Discovery. The 17-inch liquid oxygen and liquid hydrogen disconnects provide the propellant feed interface from the external tank to the orbiter main propulsion system and the three Shuttle main engines.

High Speed Optical Diagnostics in a High Pressure, GOx/RP 2 Combustor

DTIC Science & Technology

2017-07-10

Similarly, the German Aerospace Center (DLR) at Lampoldshausen conducted research on LOX/LH2 propellants in the Combustion Chamber C experimental ...facility. This single element, optically accessible liquid rocket engine test article was designed to operate up to 1,450 psi, however most research was...significant result of this work was the acquisition of data at pressures up to 2400 psi. T 3 II. Experimental Facility This testing was

Roles of Copper-Binding Proteins in Breast Cancer.

PubMed

Blockhuys, Stéphanie; Wittung-Stafshede, Pernilla

2017-04-20

Copper ions are needed in several steps of cancer progression. However, the underlying mechanisms, and involved copper-binding proteins, are mainly elusive. Since most copper ions in the body (in and outside cells) are protein-bound, it is important to investigate what copper-binding proteins participate and, for these, how they are loaded with copper by copper transport proteins. Mechanistic information for how some copper-binding proteins, such as extracellular lysyl oxidase (LOX), play roles in cancer have been elucidated but there is still much to learn from a biophysical molecular viewpoint. Here we provide a summary of copper-binding proteins and discuss ones reported to have roles in cancer. We specifically focus on how copper-binding proteins such as mediator of cell motility 1 (MEMO1), LOX, LOX-like proteins, and secreted protein acidic and rich in cysteine (SPARC) modulate breast cancer from molecular and clinical aspects. Because of the importance of copper for invasion/migration processes, which are key components of cancer metastasis, further insights into the actions of copper-binding proteins may provide new targets to combat cancer.

A study of boiling heat transfer as applied to the cooling of ball bearings in the high pressure oxygen turbopump of the space shuttle main engine

NASA Technical Reports Server (NTRS)

Schreiber, Will

1986-01-01

Two sets of ball bearings support the main shaft within the High Pressure Oxygen Turbopump (HPOTP) in the Space Shuttle Main Engine (SSME). In operation, these bearings are cooled and lubricated with high pressure liquid oxygen (LOX) flowing axially through the bearing assembly. Currently, modifications in the assembly design are being contemplated in order to enhance the lifetime of the bearings and to allow the HPOTP to operate under larger loads. An understanding of the fluid dynamics and heat transfer characteristics of the flowing LOX is necessary for the implementation of these design changes. The proposed computational model of the LOX fluid dynamics, in addition to dealing with a turbulent flow in a complex geometry, must address the complication associated with boiling and two-phase flow. The feasibility of and possible methods for modeling boiling heat transfer are considered. The theory of boiling as pertains to this particular problem is reviewed. Recommendations are given for experiments which would be necessary to establish validity for correlations needed to model boiling.

Orbital transfer rocket engine technology program: Soft wear ring seal technology

NASA Technical Reports Server (NTRS)

Lariviere, Brian W.

1992-01-01

Liquid oxygen (LOX) compatibility tests, including autogenous ignition, promoted ignition, LOX impact tests, and friction and wear tests on different PV products were conducted for several polymer materials as verification for the implementation of soft wear ring seals in advanced rocket engine turbopumps. Thermoplastics, polyimide based materials, and polyimide-imide base materials were compared for oxygen compatibility, specific wear coefficient, wear debris production, and heat dissipation mechanisms. A thermal model was generated that simulated the frictional heating input and calculated the surface temperature and temperature distribution within the seal. The predictions were compared against measured values. Heat loads in the model were varied to better match the test data and determine the difference between the measured and the calculated coefficients of friction.

Space Launch System, Core Stage, Structural Test Design and Implementation

NASA Technical Reports Server (NTRS)

Shaughnessy, Ray

2017-01-01

As part of the National Aeronautics and Space Administration's (NASA) Space Launch System (SLS) Program, engineers at NASA's Marshall Space Flight Center (MSFC) in Huntsville, Alabama are working to design, develop and implement the SLS Core Stage structural testing. The SLS will have the capability to return humans to the Moon and beyond and its first launch is scheduled for December of 2017. The SLS Core Stage consist of five major elements; Forward Skirt, Liquid Oxygen (LOX) tank, Intertank (IT), Liquid Hydrogen (LH2) tank and the Engine Section (ES). Structural Test Articles (STA) for each of these elements are being designed and produced by Boeing at Michoud Assembly Facility located in New Orleans, La. The structural test for the Core Stage STAs (LH2, LOX, IT and ES) are to be conducted by the MSFC Test Laboratory. Additionally, the MSFC Test Laboratory manages the Structural Test Equipment (STE) design and development to support the STAs. It was decided early (April 2012) in the project life that the LH2 and LOX tank STAs would require new test stands and the Engine Section and Intertank would be tested in existing facilities. This decision impacted schedules immediately because the new facilities would require Construction of Facilities (C of F) funds that require congressional approval and long lead times. The Engine Section and Intertank structural test are to be conducted in existing facilities which will limit lead times required to support the first launch of SLS. With a SLS launch date of December, 2017 Boeing had a need date for testing to be complete by September of 2017 to support flight certification requirements. The test facilities were required to be ready by October of 2016 to support test article delivery. The race was on to get the stands ready before Test Article delivery and meet the test complete date of September 2017. This paper documents the past and current design and development phases and the supporting processes, tools, and methodology for supporting the SLS Core Stage STA test stands and related STE. The paper will address key requirements, system development activities and project challenges. Additionally, the interrelationships as well as interdependencies within the SLS project will be discussed.

A dual positional specific lipoxygenase functions in the generation of flavor compounds during climacteric ripening of apple

PubMed Central

Schiller, Doreen; Contreras, Carolina; Vogt, Jörg; Dunemann, Frank; Defilippi, Bruno G; Beaudry, Randolph; Schwab, Wilfried

2015-01-01

Lipoxygenase (LOX) is an important contributor to the formation of aroma-active C6 aldehydes in apple (Malus × domestica) fruit upon tissue disruption but little is known about its role in autonomously produced aroma volatiles from intact tissue. We explored the expression of 22 putative LOX genes in apple throughout ripening, but only six LOXs were expressed in a ripening-dependent manner. Recombinant LOX1:Md:1a, LOX1:Md:1c, LOX2:Md:2a and LOX2:Md:2b proteins showed 13/9-LOX, 9-LOX, 13/9-LOX and 13-LOX activity with linoleic acid, respectively. While products of LOX1:Md:1c and LOX2:Md:2b were S-configured, LOX1:Md:1a and LOX2:Md:2a formed 13(R)-hydroperoxides as major products. Site-directed mutagenesis of Gly567 to an alanine converted the dual positional specific LOX1:Md:1a to an enzyme with a high specificity for 9(S)-hydroperoxide formation. The high expression level of the corresponding MdLOX1a gene in stored apple fruit, the genetic association with a quantitative trait locus for fruit ester and the remarkable agreement in regio- and stereoselectivity of the LOX1:Md:1a reaction with the overall LOX activity found in mature apple fruits, suggest a major physiological function of LOX1:Md:1a during climacteric ripening of apples. While LOX1:Md:1c, LOX2:Md:2a and LOX2:Md:2b may contribute to aldehyde production in immature fruit upon cell disruption our results furnish additional evidence that LOX1:Md:1a probably regulates the availability of precursors for ester production in intact fruit tissue. PMID:26504564

Comparison of pharmacokinetics between loxoprofen and its derivative with lower ulcerogenic activity, fluoro-loxoprofen.

PubMed

Yamakawa, Naoki; Suemasu, Shintaro; Watanabe, Hiroshi; Tahara, Kayoko; Tanaka, Ken-ichiro; Okamoto, Yoshinari; Ohtsuka, Masami; Maruyama, Toru; Mizushima, Tohru

2013-01-01

  We recently reported that, compared to loxoprofen (LOX, an non-steroidal anti-inflammatory drug), the LOX derivative fluoro-loxoprofen (F-LOX) is less ulcerogenic but has similar anti-inflammatory activity. Our previous in vitro studies suggested that both LOX and F-LOX are pro-drugs, the active metabolites of which are their trans-alcohol forms. In this study, we compared the pharmacokinetics of F-LOX and LOX in rats. Overall, the pharmacokinetic characteristics of F-LOX, including the formation of metabolites in vivo and in vitro, were comparable to those of LOX. However, F-LOX disappeared from the plasma more rapidly than LOX, which could potentially explain its lower ulcerogenicity. However, we showed that F-LOX produced fewer gastric lesions than LOX, even when a higher plasma concentration of F-LOX was maintained. Similar to LOX, F-LOX was readily metabolized to its trans- and cis-alcohol forms, with a higher level of the trans-alcohol form being observed after oral or intravenous administration of the drug. The preferential formation of the trans-alcohol form was also observed after incubation of F-LOX with rat liver homogenates in vitro. These results suggest that, similar to LOX, F-LOX acts as a pro-drug and that there is a metabolic system that selectively produces its active metabolite.

Crosslinked elastic fibers are necessary for low energy loss in the ascending aorta.

PubMed

Kim, Jungsil; Staiculescu, Marius Catalin; Cocciolone, Austin J; Yanagisawa, Hiromi; Mecham, Robert P; Wagenseil, Jessica E

2017-08-16

In the large arteries, it is believed that elastin provides the resistance to stretch at low pressure, while collagen provides the resistance to stretch at high pressure. It is also thought that elastin is responsible for the low energy loss observed with cyclic loading. These tenets are supported through experiments that alter component amounts through protease digestion, vessel remodeling, normal growth, or in different artery types. Genetic engineering provides the opportunity to revisit these tenets through the loss of expression of specific wall components. We used newborn mice lacking elastin (Eln -/- ) or two key proteins (lysyl oxidase, Lox -/- , or fibulin-4, Fbln4 -/- ) that are necessary for the assembly of mechanically-functional elastic fibers to investigate the contributions of elastic fibers to large artery mechanics. We determined component content and organization and quantified the nonlinear and viscoelastic mechanical behavior of Eln -/- , Lox -/- , and Fbln4 -/- ascending aorta and their respective controls. We confirmed that the lack of elastin, fibulin-4, or lysyl oxidase leads to absent or highly fragmented elastic fibers in the aortic wall and a 56-97% decrease in crosslinked elastin amounts. We found that the resistance to stretch at low pressure is decreased only in Eln -/- aorta, confirming the role of elastin in the nonlinear mechanical behavior of the aortic wall. Dissipated energy with cyclic loading and unloading is increased 53-387% in Eln -/- , Lox -/- , and Fbln4 -/- aorta, indicating that not only elastin, but properly assembled and crosslinked elastic fibers, are necessary for low energy loss in the aorta. Copyright © 2017 Elsevier Ltd. All rights reserved.

A logistics and potential hazard study of propellant systems for a Saturn 5 derived heavy lift (three-stage core) launch vehicle

NASA Technical Reports Server (NTRS)

Whitney, E. Dow

1992-01-01

The Bush Administration has directed NASA to prepare for a return to the Moon and on to Mars - the Space Exploration Initiative. To meet this directive, powerful rocket boosters will be required in order to lift payloads that may reach the half-million pound range into low earth orbit. In this report an analysis is presented on logistics and potential hazards of the propellant systems envisioned for future Saturn 5 derived heavy lift launch vehicles. In discussing propellant logistics, particular attention has been given to possible problems associated with procurement, transportation, and storage of RP-1, HL2, and LOX, the heavy lift launch vehicle propellants. Current LOX producing facilities will need to be expanded and propellant storage and some support facilities will require relocation if current Launch Pads 39A and/or 39B are to be used for future heavy noise-abatement measures. Included in the report is a discussion of suggested additional studies, primarily economic and environmental, which should be undertaken in support of the goals of the Space Exploration Initiative.

Test Analysis Correlation of the Single Stringer Bending Tests for the Space Shuttle ET-137 Intertank Stringer Crack Investigation

NASA Technical Reports Server (NTRS)

Phillips, Dawn R.; Saxon, Joseph B.; Wingate, Robert J.

2012-01-01

On November 5, 2010, Space Shuttle mission STS-133 was scrubbed due to a hydrogen leak at the Ground Umbilical Carrier Plate (GUCP). After the scrub, a crack in the foam thermal protection system (TPS) was observed on the External Tank (ET) near the interface between the liquid oxygen (LOX) tank and the Intertank. When the damaged foam was removed, two 9-in. long cracks were found on the feet of Intertank stringer S7-2, and the stringer failure was the cause of the TPS crack. An investigation was conducted to determine the root cause of the cracks, establish a remedy/repair for the stringers, and provide flight rationale for the damaged tank, ET-137. The Space Transportation System (STS) Super Lightweight ET (SLWT) is comprised of two propellant tanks (an aft liquid hydrogen (LH2) tank and a forward LOX tank) and an Intertank. The Intertank serves as the structural connection between the two propellant tanks and also functions to receive and distribute all thrust loads from the solid rocket boosters . The Intertank is a stiffened cylinder structure consisting of eight mechanically joined panels (two integrally-stiffened, machined thrust panels to react the booster loads and six stringer-stiffened skin panels). There are one main ring frame, four intermediate ring frames, and forward and aft flange chords that mate to the respective propellant tanks.. The skin/stringer panels utilize external hat-section stringers that are mechanically attached with rivets along most of their length and with specialty fasteners, such as GP Lockbolts and Hi-Loks, at the forward and aft ends where the stringers attach to the flange chords. During the STS-133 Intertank stringer crack investigation, cracks were found on a total of five stringers. All of the cracks were at the LOX end, in the feet of the stringers, and near the forward fasteners (GP Lockbolts). Video of tanking for the November 5 launch attempt was used to determine that the TPS failure, and thus the stringer failure, occurred as the LOX liquid level crossed the LOX tank / Intertank interface ring frame. Hence, cryogenically-induced displacements were suspected as a contributing cause of the stringer cracks. To study the behavior of Intertank stringers subjected to similar displacements, static load tests of individual stringers, colloquially known as "single stringer bending tests" were performed. Approximately thirty stringers were tested, many of which were cut from the partially completed Intertank for what would have been ET-139. In addition to the tests, finite element (FE) analyses of the test configuration were also performed. In this paper, the FE analyses and test-analysis correlation for stringer test S6-8 are presented. Stringer S6-8 is a "short chord" configuration with no doubler panels.

A novel proteolytic processing of prolysyl oxidase

PubMed Central

Atsawasuwan, Phimon; Mochida, Yoshiyuki; Katafuchi, Michitsuna; Tokutomi, Kentaro; Mocanu, Viorel; Parker, Carol E.; Yamauchi, Mitsuo

2012-01-01

Lysyl oxidase (LOX) is an amine oxidase that is critical for the stability of connective tissues. The secreted proLOX is enzymatically quiescent and is activated through proteolytic cleavage between residue Gly162 and Asp163 (residue numbers according to the mouse LOX) by bone morphogenetic protein (BMP)-1 gene products. Here we report a novel processing of proLOX identified in vitro and in vivo. Two forms of mature LOX were identified and characterized by their immunoreactivity to specific antibodies, amine oxidase activity and mass spectrometry. One form was identified as a well characterized BMP-1 processed LOX protein. Another was found to be a truncated form of LOX (tLOX) resulting from the cleavage at the carboxy terminus of Arg192. The tLOX still appeared to retain amine oxidase activity. The results from the proLOX gene deletion and mutation experiments indicated that the processing occurs independent of the cleavage of proLOX by BMP-1 gene products and likely requires the presence of LOX propeptide. These results indicate that proLOX could be processed by two different mechanisms producing two forms of active LOX. PMID:21591931

Cre/lox-based multiple markerless gene disruption in the genome of the extreme thermophile Thermus thermophilus.

PubMed

Togawa, Yoichiro; Nunoshiba, Tatsuo; Hiratsu, Keiichiro

2018-02-01

Markerless gene-disruption technology is particularly useful for effective genetic analyses of Thermus thermophilus (T. thermophilus), which have a limited number of selectable markers. In an attempt to develop a novel system for the markerless disruption of genes in T. thermophilus, we applied a Cre/lox system to construct a triple gene disruptant. To achieve this, we constructed two genetic tools, a loxP-htk-loxP cassette and cre-expressing plasmid, pSH-Cre, for gene disruption and removal of the selectable marker by Cre-mediated recombination. We found that the Cre/lox system was compatible with the proliferation of the T. thermophilus HB27 strain at the lowest growth temperature (50 °C), and thus succeeded in establishing a triple gene disruptant, the (∆TTC1454::loxP, ∆TTC1535KpnI::loxP, ∆TTC1576::loxP) strain, without leaving behind a selectable marker. During the process of the sequential disruption of multiple genes, we observed the undesired deletion and inversion of the chromosomal region between multiple loxP sites that were induced by Cre-mediated recombination. Therefore, we examined the effects of a lox66-htk-lox71 cassette by exploiting the mutant lox sites, lox66 and lox71, instead of native loxP sites. We successfully constructed a (∆TTC1535::lox72, ∆TTC1537::lox72) double gene disruptant without inducing the undesired deletion of the 0.7-kbp region between the two directly oriented lox72 sites created by the Cre-mediated recombination of the lox66-htk-lox71 cassette. This is the first demonstration of a Cre/lox system being applicable to extreme thermophiles in a genetic manipulation. Our results indicate that this system is a powerful tool for multiple markerless gene disruption in T. thermophilus.

Two closely related members of Arabidopsis 13-lipoxygenases (13-LOXs), LOX3 and LOX4, reveal distinct functions in response to plant-parasitic nematode infection.

PubMed

Ozalvo, Rachel; Cabrera, Javier; Escobar, Carolina; Christensen, Shawn A; Borrego, Eli J; Kolomiets, Michael V; Castresana, Carmen; Iberkleid, Ionit; Brown Horowitz, Sigal

2014-05-01

The responses of two closely related members of Arabidopsis 13-lipoxygenases (13-LOXs), LOX3 and LOX4, to infection by the sedentary nematodes root-knot nematode (Meloidogyne javanica) and cyst nematode (Heterodera schachtii) were analysed in transgenic Arabidopsis seedlings. The tissue localization of LOX3 and LOX4 gene expression using β-glucuronidase (GUS) reporter gene constructs showed local induction of LOX3 expression when second-stage juveniles reached the vascular bundle and during the early stages of plant-nematode interaction through gall and syncytia formation. Thin sections of nematode-infested knots indicated LOX3 expression in mature giant cells, and high expression in neighbouring cells and those surrounding the female body. LOX4 promoter was also activated by nematode infection, although the GUS signal weakened as infection and disease progressed. Homozygous insertion mutants lacking LOX3 were less susceptible than wild-type plants to root-knot nematode infection, as reflected by a decrease in female counts. Conversely, deficiency in LOX4 function led to a marked increase in females and egg mass number and in the female to male ratio of M. javanica and H. schachtii, respectively. The susceptibility of lox4 mutants was accompanied by increased expression of allene oxide synthase, allene oxide cyclase and ethylene-responsive transcription factor 4, and the accumulation of jasmonic acid, measured in the roots of lox4 mutants. This response was not found in lox3 mutants. Taken together, our results reveal that LOX4 and LOX3 interfere differentially with distinct metabolic and signalling pathways, and that LOX4 plays a major role in controlling plant defence against nematode infection. © 2013 BSPP AND JOHN WILEY & SONS LTD.

A novel proteolytic processing of prolysyl oxidase.

PubMed

Atsawasuwan, Phimon; Mochida, Yoshiyuki; Katafuchi, Michitsuna; Tokutomi, Kentaro; Mocanu, Viorel; Parker, Carol E; Yamauchi, Mitsuo

2011-01-01

Lysyl oxidase (LOX) is an amine oxidase that is critical for the stability of connective tissues. The secreted proLOX is enzymatically quiescent and is activated through proteolytic cleavage between residues Gly(162) and Asp(163) (residue numbers according to the mouse LOX) by bone morphogenetic protein (BMP)-1 gene products. Here we report a novel processing of proLOX identified in vitro and in vivo. Two forms of mature LOX were identified and characterized by their immunoreactivity to specific antibodies, amine oxidase activity, and mass spectrometry. One form was identified as a well-characterized BMP-1 processed LOX protein. Another was found to be a truncated form of LOX resulting from the cleavage at the carboxy terminus of Arg(192). The truncated form of LOX still appeared to retain amine oxidase activity. The results from the proLOX gene deletion and mutation experiments indicated that the processing occurs independent of the cleavage of proLOX by BMP-1 gene products and likely requires the presence of LOX propeptide. These results indicate that proLOX could be processed by two different mechanisms producing two forms of active LOX.

LOX is a novel mitotic spindle-associated protein essential for mitosis.

PubMed

Boufraqech, Myriem; Wei, Darmood; Weyemi, Urbain; Zhang, Lisa; Quezado, Martha; Kalab, Petr; Kebebew, Electron

2016-05-17

LOX regulates cancer progression in a variety of human malignancies. It is overexpressed in aggressive cancers and higher expression of LOX is associated with higher cancer mortality. Here, we report a new function of LOX in mitosis. We show that LOX co-localizes to mitotic spindles from metaphase to telophase, and p-H3(Ser10)-positive cells harbor strong LOX staining. Further, purification of mitotic spindles from synchronized cells show that LOX fails to bind to microtubules in the presence of nocodazole, whereas paclitaxel treated samples showed enrichment in LOX expression, suggesting that LOX binds to stabilized microtubules. LOX knockdown leads to G2/M phase arrest; reduced p-H3(Ser10), cyclin B1, CDK1, and Aurora B. Moreover, LOX knockdown significantly increased sensitivity of cancer cells to chemotherapeutic agents that target microtubules. Our findings suggest that LOX has a role in cancer cell mitosis and may be targeted to enhance the activity of microtubule inhibitors for cancer therapy.

Lox/Gox related failures during Space Shuttle Main Engine development

NASA Technical Reports Server (NTRS)

Cataldo, C. E.

1981-01-01

Specific rocket engine hardware and test facility system failures are described which were caused by high pressure liquid and/or gaseous oxygen reactions. The failures were encountered during the development and testing of the space shuttle main engine. Failure mechanisms are discussed as well as corrective actions taken to prevent or reduce the potential of future failures.

Composite Load Spectra for Select Space Propulsion Structural Components

NASA Technical Reports Server (NTRS)

Ho, Hing W.; Newell, James F.

1994-01-01

Generic load models are described with multiple levels of progressive sophistication to simulate the composite (combined) load spectra (CLS) that are induced in space propulsion system components, representative of Space Shuttle Main Engines (SSME), such as transfer ducts, turbine blades and liquid oxygen (LOX) posts. These generic (coupled) models combine the deterministic models for composite load dynamic, acoustic, high-pressure and high rotational speed, etc., load simulation using statistically varying coefficients. These coefficients are then determined using advanced probabilistic simulation methods with and without strategically selected experimental data. The entire simulation process is included in a CLS computer code. Applications of the computer code to various components in conjunction with the PSAM (Probabilistic Structural Analysis Method) to perform probabilistic load evaluation and life prediction evaluations are also described to illustrate the effectiveness of the coupled model approach.

Air Launch: Examining Performance Potential of Various Configurations and Growth Options

NASA Technical Reports Server (NTRS)

Waters, Eric D.; Creech, Dennis M.; Philips, Alan D.

2013-01-01

The Advanced Concepts Office at NASA's George C. Marshall Space Flight Center conducted a high-level analysis of various air launch vehicle configurations, objectively determining maximum launch vehicle payload while considering carrier aircraft capabilities and given dimensional constraints. With the renewed interest in aerial launch of low-earth orbit payloads, referenced by programs such as Stratolaunch and Spaceship2, there exists a need to qualify the boundaries of the trade space, identify performance envelopes, and understand advantages and limiting factors of designing for maximum payload capability. Using the NASA/DARPA Horizontal Launch Study (HLS) Point Design 2 (PD-2) as a pointof- departure configuration, two independent design actions were undertaken. Both designs utilized a Boeing 747-400F as the carrier aircraft, LOX/RP-1 first stage and LOX/LH2 second stage. Each design was sized to meet dimensional and mass constraints while optimizing propellant loads and stage delta V splits. All concepts, when fully loaded, exceeded the allowable Gross Takeoff Weight (GTOW) of the aircraft platform. This excess mass was evaluated as propellant/fuel offload available for a potential in-flight propellant loading scenario. Results indicate many advantages such as payload delivery of approximately 47,000 lbm and significant mission flexibility including variable launch site inclination and launch window. However, in-flight cryogenic fluid transfer and carrier aircraft platform integration are substantial technical hurdles to the realization of such a system configuration.

Cucumber Cotyledon Lipoxygenase during Postgerminative Growth. Its Expression and Action on Lipid Bodies

PubMed Central

Matsui, Kenji; Hijiya, Kohko; Tabuchi, Yutaka; Kajiwara, Tadahiko

1999-01-01

In cucumber (Cucumis sativus), high lipoxygenase-1 (LOX-1) activity has been detected in the soluble fraction prepared from cotyledons of germinating seeds, and the involvement of this enzyme in lipid turnover has been suggested (K. Matsui, M. Irie, T. Kajiwara, A. Hatanaka [1992] Plant Sci 85: 23–32; I. Fuessner, C. Wasternack, H. Kindl, H. Kühn [1995] Proc Natl Acad Sci USA 92: 11849–11853). In this study we have investigated the expression of the gene lox-1, corresponding to the LOX-1 enzyme. LOX-1 expression is highly coordinated with that of a typical glyoxysomal enzyme, isocitrate lyase, during the postgerminative stage of cotyledon development. In contrast, although icl transcripts accumulated in tissue during in vitro senescence, no accumulation of lox-1 mRNA could be observed, suggesting that lox-1 plays a specialized role in fat mobilization. LOX-1 is also known to be a major lipid body protein. The partial peptide sequences of purified LOX-1 and lipid body LOX-1 entirely coincided with that deduced from the lox-1 cDNA sequence. The data strongly suggest that LOX-1 and lipid body LOX-1 are derived from a single gene and that LOX-1 can exist both in the cytosol and on the lipid bodies. We constructed an in vitro oxygenation system to address the mechanism of this dual localization and to investigate the action of LOX-1 on lipids in the lipid bodies. LOX-1 cannot act on the lipids in intact lipid bodies, although degradation of lipid body proteins, either during seedling growth or by treatment with trypsin, allows lipid bodies to become susceptible to LOX-1. We discuss the role of LOX-1 in fat mobilization and its mechanism of action. PMID:10198086

77 FR 21619 - Office of Commercial Space Transportation; Notice of Intent To Prepare an Environmental Impact...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-04-10

... rocket propellant-1 or refined petroleum-1 (RP-1), as propellants to carry payloads into orbit. The... tank with a maximum propellant (RP-1 and LOX) load of approximately 6,900 gallons. As part of the... processing-hangar, a launch pad and stand with its associated flame duct, propellant storage and handling...

Milestone report TCTP application to the SSME hydrogen system analysis

NASA Technical Reports Server (NTRS)

Richards, J. S.

1975-01-01

The Transient Cryogen Transfer Computer Program (TCTP) developed and verified for LOX systems by analyses of Skylab S-1B stage loading data from John F. Kennedy Space Center launches was extended to include hydrogen as the working fluid. The feasibility of incorporating TCTP into the space shuttle main engine dynamic model was studied. The program applications are documented.

Downregulation of Lysyl Oxidase Protects Retinal Endothelial Cells From High Glucose-Induced Apoptosis.

PubMed

Kim, Dongjoon; Mecham, Robert P; Trackman, Philip C; Roy, Sayon

2017-05-01

To investigate the effect of reducing high glucose (HG)-induced lysyl oxidase (LOX) overexpression and increased activity on retinal endothelial cell apoptosis. Rat retinal endothelial cells (RRECs) were grown in normal (N) or HG (30 mM glucose) medium for 7 days. In parallel, RRECs were grown in HG medium and transfected with LOX small interfering RNA (siRNA), scrambled siRNA as control, or exposed to β-aminopropionitrile (BAPN), a LOX inhibitor. LOX expression, AKT activation, and caspase-3 activity were determined by Western blot (WB) analysis and apoptosis by differential dye staining assay. Moreover, to determine whether diabetes-induced LOX overexpression alters AKT activation and promotes apoptosis, changes in LOX expression, AKT phosphorylation, caspase-3 activation, and Bax expression were assessed in retinas of streptozotocin (STZ)-induced diabetic mice and LOX heterozygous knockout (LOX+/-) mice. WB analysis indicated significant LOX overexpression and reduced AKT activation under HG condition in RRECs. Interestingly, when cells grown in HG were transfected with LOX siRNA or exposed to BAPN, the number of apoptotic cells was significantly decreased concomitant with increased AKT phosphorylation. Diabetic mouse retinas exhibited LOX overexpression, decreased AKT phosphorylation, and increased Bax and caspase-3 activation compared to values in nondiabetic mice. In LOX+/- mice, reduced LOX levels were observed with increased AKT activity, and reduced Bax and caspase-3 activity. Furthermore, decreased levels of LOX in the LOX+/- mice was protective against diabetes-induced apoptosis. Findings from this study indicate that preventing LOX overexpression may be protective against HG-induced apoptosis in retinal vascular cells associated with diabetic retinopathy.

Two closely related members of Arabidopsis 13-LOXs, LOX3 and LOX4, reveal distinct functions in response to plantparasitic nematode infection

USDA-ARS?s Scientific Manuscript database

The responses of two closely related members of Arabidopsis 13-lipoxygenase (13-LOX), LOX3 and LOX4, to infection by the sedentary nematodes root-knot nematode (Meloidogyne javanica) and cyst nematode (Heterodera schachtii) were analyzed in transgenic Arabidopsis seedlings. Tissue localization of LO...

5-Lipoxygenase-activating protein rescues activity of 5-lipoxygenase mutations that delay nuclear membrane association and disrupt product formation.

PubMed

Gerstmeier, Jana; Newcomer, Marcia E; Dennhardt, Sophie; Romp, Erik; Fischer, Jana; Werz, Oliver; Garscha, Ulrike

2016-05-01

Leukotrienes (LTs) are proinflammatory lipid mediators formed from arachidonic acid in a 2-step reaction catalyzed by 5-lipoxygenase (5-LOX) requiring the formation of 5-HPETE [5(S)-hydroperoxy-6-trans-8,11,14-cis-eicosatetraenoic acid] and its subsequent transformation to LTA4 5-LOX is thought to receive arachidonic acid from the nuclear membrane-embedded 5-LOX-activating protein (FLAP). The crystal structure of 5-LOX revealed an active site concealed by F177 and Y181 (FY cork). We examined the influence of the FY cork on 5-LOX activity and membrane binding in HEK293 cells in the absence and presence of FLAP. Uncapping the 5-LOX active site by mutation of F177 and/or Y181 to alanine (5-LOX-F177A, 5-LOX-Y181A, 5-LOX-F177/Y181A) resulted in delayed and diminished 5-LOX membrane association in A23187-stimulated cells. For 5-LOX-F177A and 5-LOX-F177/Y181A, formation of 5-LOX products was dramatically reduced relative to 5-LOX-wild type (wt). Strikingly, coexpression of FLAP in A23187-activated HEK293 cells effectively restored formation of 5-H(p)ETE (5-hydroxy- and 5-peroxy-6-trans-8,11,14-cis-eicosatetraenoic acid) by these same 5-LOX mutants (≈60-70% 5-LOX-wt levels) but not of LTA4 hydrolysis products. Yet 5-LOX-Y181A generated 5-H(p)ETE at levels comparable to 5-LOX-wt but reduced LTA4 hydrolysis products. Coexpression of FLAP partially restored LTA4 hydrolysis product formation by 5-LOX-Y181A. Together, the data suggest that the concealed FY cork impacts membrane association and that FLAP may help shield an uncapped active site.-Gerstmeier, J., Newcomer, M. E., Dennhardt, S., Romp, E., Fischer, J., Werz, O., Garscha, U. 5-Lipoxygenase-activating protein rescues activity of 5-lipoxygenase mutations that delay nuclear membrane association and disrupt product formation. © FASEB.

Crystal Structure of Manganese Lipoxygenase of the Rice Blast Fungus Magnaporthe oryzae*

PubMed Central

Wennman, Anneli; Oliw, Ernst H.; Karkehabadi, Saeid; Chen, Yang

2016-01-01

Lipoxygenases (LOX) are non-heme metal enzymes, which oxidize polyunsaturated fatty acids to hydroperoxides. All LOX belong to the same gene family, and they are widely distributed. LOX of animals, plants, and prokaryotes contain iron as the catalytic metal, whereas fungi express LOX with iron or with manganese. Little is known about metal selection by LOX and the adjustment of the redox potentials of their protein-bound catalytic metals. Thirteen three-dimensional structures of animal, plant, and prokaryotic FeLOX are available, but none of MnLOX. The MnLOX of the most important plant pathogen, the rice blast fungus Magnaporthe oryzae (Mo), was expressed in Pichia pastoris. Mo-MnLOX was deglycosylated, purified to homogeneity, and subjected to crystal screening and x-ray diffraction. The structure was solved by sulfur and manganese single wavelength anomalous dispersion to a resolution of 2.0 Å. The manganese coordinating sphere is similar to iron ligands of coral 8R-LOX and soybean LOX-1 but is not overlapping. The Asn-473 is positioned on a short loop (Asn-Gln-Gly-Glu-Pro) instead of an α-helix and forms hydrogen bonds with Gln-281. Comparison with FeLOX suggests that Phe-332 and Phe-525 might contribute to the unique suprafacial hydrogen abstraction and oxygenation mechanism of Mo-MnLOX by controlling oxygen access to the pentadiene radical. Modeling suggests that Arg-525 is positioned close to Arg-182 of 8R-LOX, and both residues likely tether the carboxylate group of the substrate. An oxygen channel could not be identified. We conclude that Mo-MnLOX illustrates a partly unique variation of the structural theme of FeLOX. PMID:26783260

Cannabidiol-2',6'-dimethyl ether, a cannabidiol derivative, is a highly potent and selective 15-lipoxygenase inhibitor.

PubMed

Takeda, Shuso; Usami, Noriyuki; Yamamoto, Ikuo; Watanabe, Kazuhito

2009-08-01

The inhibitory effect of nordihydroguaiaretic acid (NDGA) (a nonselective lipoxygenase (LOX) inhibitor)-mediated 15-LOX inhibition has been reported to be affected by modification of its catechol ring, such as methylation of the hydroxyl group. Cannabidiol (CBD), one of the major components of marijuana, is known to inhibit LOX activity. Based on the phenomenon observed in NDGA, we investigated whether or not methylation of CBD affects its inhibitory potential against 15-LOX, because CBD contains a resorcinol ring, which is an isomer of catechol. Although CBD inhibited 15-LOX activity with an IC(50) value (50% inhibition concentration) of 2.56 microM, its monomethylated and dimethylated derivatives, CBD-2'-monomethyl ether and CBD-2',6'-dimethyl ether (CBDD), inhibited 15-LOX activity more strongly than CBD. The number of methyl groups in the resorcinol moiety of CBD (as a prototype) appears to be a key determinant for potency and selectivity in inhibition of 15-LOX. The IC(50) value of 15-LOX inhibition by CBDD is 0.28 microM, and the inhibition selectivity for 15-LOX (i.e., the 5-LOX/15-LOX ratio of IC(50) values) is more than 700. Among LOX isoforms, 15-LOX is known to be able to oxygenate cholesterol esters in the low-density lipoprotein (LDL) particle (i.e., the formation of oxidized LDL). Thus, 15-LOX is suggested to be involved in development of atherosclerosis, and CBDD may be a useful prototype for producing medicines for atherosclerosis.

Dysregulation of Lysyl Oxidase Expression in Lesions and Endometrium of Women With Endometriosis

PubMed Central

Ruiz, Lynnette A.; Báez-Vega, Perla M.; Ruiz, Abigail; Peterse, Daniëlle P.; Monteiro, Janice B.; Bracero, Nabal; Beauchamp, Pedro; Fazleabas, Asgerally T.; Flores, Idhaliz

2015-01-01

Lysyl oxidases (LOXs) are enzymes involved in collagen deposition, extracellular membrane remodeling, and invasive/metastatic potential. Previous studies reveal an association of LOXs and endometriosis. We aimed to identify the mechanisms activated by upregulation of lysyl oxidases (LOX) in endometriotic cells and tissues. We hypothesized that LOX plays a role in endometriosis by promoting invasiveness and epithelial to mesenchymal transition (EMT). Methods: The LOX protein expression levels were measured by immunohistochemistry in lesions and endometrium on a tissue microarray (TMA) and in endometrial biopsies from patients and controls during the window of implantation (WOI). Estradiol regulation of LOX expression was determined by quantitative polymerase chain reaction (qPCR). Proliferation, invasion, and migration assays were performed in epithelial (endometrial epithelial cell), endometrial (human endometrial stromal cell), and endometriotic cell lines (ECL and 12Z). Pathway-focused multiplex qPCR was used to determine transcriptome changes due to LOX overexpression. Results: LOX protein was differentially expressed in ovarian versus peritoneal lesions. During WOI, LOX levels were higher in luminal epithelium of patients with endometriosis-associated infertility compared to controls. Invasive epithelial cell lines expressed higher levels of LOX than noninvasive ones. Transfection of LOX into noninvasive epithelial cells increased their migration in an LOX inhibitor-sensitive manner. Overexpression of LOX did not fully induce EMT but the expression of genes related to fibrosis and extracellular matrix remodeling were dysregulated. Conclusions: This study documents that expression of LOX is differentially regulated in endometriotic lesions and endometrium. A role for LOX in mediating proliferation, migration, and invasion of endometrial and endometriotic cells was observed, which may be implicated in the establishment and progression of endometriotic lesions. PMID:25963914

Lectin-like oxidized low-density lipoprotein receptor (LOX-1) in sickle cell disease vasculopathy

PubMed Central

Chen, Mingyi; Qiu, Hong; Lin, Xin; Nam, David; Ogbu-Nwobodo, Lucy; Archibald, Hannah; Joslin, Amelia; Wun, Ted; Sawamura, Tatsuya; Green, Ralph

2017-01-01

Lectin-like oxidized low-density lipoprotein (LDL) receptor-1 (LOX-1) is an endothelial receptor for oxidized LDL. Increased expression of LOX-1 has been demonstrated in atherosclerotic lesions and diabetic vasculopathy. In this study, we investigate the expression of LOX-1 receptor in sickle cell disease (SCD) vasculopathy. Expression of LOX-1 in brain vascular endothelium is markedly increased and LOX-1 gene expression is upregulated in cultured human brain microvascular endothelial cells by incubation with SCD erythrocytes. Also, the level of circulating soluble LOX-1 concentration is elevated in the plasma of SCD patients. Increased LOX-1 expression in endothelial cells is potentially involved in the pathogenesis of SCD vasculopathy. Soluble LOX-1 concentration in SCD may provide a novel biomarker for risk stratification of sickle cell vascular complications. PMID:27519944

Crystal structure of human lysyl oxidase-like 2 (hLOXL2) in a precursor state.

PubMed

Zhang, Xi; Wang, Qifan; Wu, Jianping; Wang, Jiawei; Shi, Yigong; Liu, Minhao

2018-04-10

Lysyl oxidases (LOXs), a type of copper- and lysyl tyrosylquinone (LTQ) -dependent amine oxidase, catalyze the oxidative deamination of lysine residues of extracellular matrix (ECM) proteins such as elastins and collagens and generate aldehyde groups. The oxidative deamination of lysine represents the foundational step for the cross-linking of elastin and collagen and thus is crucial for ECM modeling. Despite their physiological significance, the structure of this important family of enzymes remains elusive. Here we report the crystal structure of human lysyl oxidase-like 2 (hLOXL2) at 2.4-Å resolution. Unexpectedly, the copper-binding site of hLOXL2 is occupied by zinc, which blocks LTQ generation and the enzymatic activity of hLOXL2 in our in vitro assay. Biochemical analysis confirms that copper loading robustly activates hLOXL2 and supports LTQ formation. Furthermore, the LTQ precursor residues in the structure are distanced by 16.6 Å, corroborating the notion that the present structure may represent a precursor state and that pronounced conformational rearrangements would be required for protein activation. The structure presented here establishes an important foundation for understanding the structure-function relationship of LOX proteins and will facilitate LOX-targeting drug discovery. Copyright © 2018 the Author(s). Published by PNAS.

Conceptual Design, Feasibility and Payoff Analysis of a Third Stage for EELV

DTIC Science & Technology

2014-06-01

the overall vehicle architecture, identifying locations for modification or stage shape. Trade studies of various propellant types (LOX/ LH2 , LOX/RP...or stage shape. Trade studies of various propellant types (LOX/ LH2 , LOX/RP, LOX/methane, hydrazine monopropellant) were included in the analysis...Geostationary transfer orbit Isp = Specific impulse LEO = Low Earth Orbit LOX = Liquid oxygen LH2 = Liquid hydrogen POST = Program to

S.S.T.O. performance assessment with in-flight lox collection

NASA Astrophysics Data System (ADS)

Vandenkerckhove, J.; Czysz, P.

1995-10-01

Much attention has recently been given, up to harware development to in-flight oxygen collection as a means to improve considerably the performance of both TSTO & SSTO vehicles. A first assessment suggests that it permits simultaneously to improve much both gross take-off weight (by more than 30%) & dry weight (by more than 15%) of an SSTO and to lower significantly the Mach number of transition scramjet → rocket, from 15 down below 10, thereby reducing dramatically the programmatic development risks. After having compared in-flight lox collection with other SSTO concepts, this paper provides a tentative assessment of the performance of SSTO vehicles taking advantage of it, in particular their sensitivity to changes in system characteristics such as transition Mach number, vehicle slenderness (i.e. Küchemann's parameter τ) or planform loading at take-off and in collection characteristics, in particular collection ratio & specific collection plant weight.

High-Payoff Space Transportation Design Approach with a Technology Integration Strategy

NASA Technical Reports Server (NTRS)

McCleskey, C. M.; Rhodes, R. E.; Chen, T.; Robinson, J.

2011-01-01

A general architectural design sequence is described to create a highly efficient, operable, and supportable design that achieves an affordable, repeatable, and sustainable transportation function. The paper covers the following aspects of this approach in more detail: (1) vehicle architectural concept considerations (including important strategies for greater reusability); (2) vehicle element propulsion system packaging considerations; (3) vehicle element functional definition; (4) external ground servicing and access considerations; and, (5) simplified guidance, navigation, flight control and avionics communications considerations. Additionally, a technology integration strategy is forwarded that includes: (a) ground and flight test prior to production commitments; (b) parallel stage propellant storage, such as concentric-nested tanks; (c) high thrust, LOX-rich, LOX-cooled first stage earth-to-orbit main engine; (d) non-toxic, day-of-launch-loaded propellants for upper stages and in-space propulsion; (e) electric propulsion and aero stage control.

Marshall Space Flight Center Autumn 2005

NASA Technical Reports Server (NTRS)

Allen, Mike; Clar, Harry E.

2006-01-01

The East Test Area at Marshall Space Flight Center has five major test stands, each of which has two or more test positions, not counting the SSME and RD-180 engine test facilities in the West Test Area. These research and development facilities are capable of testing high pressure pumps, both fuel and oxidizer, injectors, chambers and sea-level engine assemblies, as well as simulating deep space environments in the 12, 15 and 20 foot vacuum chambers. Liquid propellant capabilities are high pressure hydrogen (liquid and gas), methane (liquid and gas), and RP-1 and high pressure LOX. Solid propellant capability includes thrust measurement and firing capability up to 1/6 scale Shuttle SRB segment. In the past six months MSFC supported multiple space access and exploration programs in the previous six months. Major programs were Space Exploration, Shuttle External Tank research, Reusable Solid Rocket Motor (RSRM) development, as well as research programs for NASA and other customers. At Test Stand 115 monopropellant ignition testing was conducted on one position. At the second position multiple ignition/variable burn time cycles were conducted on Vacuum Plasma Spatter (VPS) coated injectors. Each injector received fifty cycles; the propellants were LOX Hydrogen and the ignition source was TEA. Following completion of the monopropellant test series the stand was reconfigured to support ignition testing on a LOX Methane injector system. At TS 116 a thrust stand used to test Booster Separation Motors from the Shuttle SRB system was disassembled and moved from Chemical Systems Division s Coyote Canyon plant to MSFC. The stand was reassembled and readied for BSM testing. Also, a series of tests was run on a Pratt & Whitney Rocketdyne Low Element Density (LED) injector engine. The propellants for this engine are LOX and LH2. At TS 300 the 20 foot vacuum chamber was configured to support hydrogen testing in the Multipurpose Hydrogen Test Bed (MHTB) test article. This testing, which went 24/7 for fourteen consecutive days, demonstrated long duration storage methods intended to minimize losses of propellant in support of the Space Exploration Initiative. The facility is being converted to support similar research using liquid methane. The 12 foot chamber at TS 300 was used to create ascent profiles (both heat and altitude effects) for foam panel testing in support of the Shuttle External Tank program. At TS 500, one position was in build-up to support ATK Thiokol research into the gas dynamics associated with high pressure flow across the propellant joint in segmented solid rocket motors. The testing involves flowing high pressure gas through a 24 motor case. Initial tests will be conducted with simulated aluminum grain, followed by tests using actual propellant. The second position at TS 500 has been in build-up for testing a LOX methane thruster manufactured by KT Engineering. At the Solid Propulsion Test Area (SPTA), the first dual segment 24 solid rocket motor was fired for ATK Thiokol in support of the RSRM program. A new axial thrust measurement stand was designed and fabricated for this testing. Real Time Radiography (RTR) will be deployed to examine nozzle erosion on the next dual segment motor.

The determination of equivalent bearing loading for the BSMT that simulate SSME high pressure oxidizer turbopump conditions using the SHABERTH/SINDA computer programs

NASA Technical Reports Server (NTRS)

Mcdonald, Gary H.

1987-01-01

The MSFC bearing seal material tester (BSMT) can be used to evaluate the SSME high pressure oxygen turbopump (HPOTP) bearing performance. The four HPOTP bearings have both an imposed radial and axial load. These radial and axial loads are caused by the HPOTP's shaft, main impeller, preburner impeller, turbine and by the LOX coolant flow through the bearings, respectively. These loads coupled with bearing geometry and operating speed can define bearing contact angle, contact Hertz stress, and heat generation rates. The BSMT has the capability of operating at HPOTP shaft speeds, provide proper coolant flowrates but can only apply an axial load. Due to the inability to operate the bearings in the BSMT with an applied radial load, it is important to develop an equivalency between the applied axial loads and the actual HPOTP loadings. A shaft-bearing-thermal computer code (SHABERTH/SINDA) is used to simulate the BSMT bearing-shaft geometry and thermal-fluid operating conditions.

Melanocortin-3 receptors expressed in Nkx2.1(+ve) neurons are sufficient for controlling appetitive responses to hypocaloric conditioning

PubMed Central

Girardet, Clémence; Mavrikaki, Maria M.; Stevens, Joseph R.; Miller, Courtney A.; Marks, Daniel L.; Butler, Andrew A.

2017-01-01

Melanocortin-3 receptors (MC3R) have a contextual role in appetite control that is amplified with hypocaloric conditioning. C57BL/6J (B6) mice subjected to hypocaloric feeding schedules (HFS) exhibit compulsive behavioral responses involving food anticipatory activity (FAA) and caloric loading following food access. These homeostatic responses to calorie-poor environs are attenuated in B6 mice in which Mc3r transcription is suppressed by a lox-stop-lox sequence in the 5’UTR (Mc3rTB/TB). Here, we report that optimization of caloric loading in B6 mice subject to HFS, characterized by increased meal size and duration, is not observed in Mc3rTB/TB mice. Analysis of hypothalamic and neuroendocrine responses to HFS throughout the light-dark cycle suggests uncoupling of hypothalamic responses involving appetite-stimulating fasting-responsive hypothalamic neurons expressing agouti-related peptide (AgRP) and neuropeptide Y (Npy). Rescuing Mc3rs expression in Nkx2.1(+ve) neurons is sufficient to restore normal hypothalamic responses to negative energy balance. In addition, Mc3rs expressed in Nkx2.1(+ve) neurons are also sufficient to restore FAA and caloric loading of B6 mice subjected to HFS. In summary, MC3Rs expressed in Nkx2.1(+ve) neurons are sufficient to coordinate hypothalamic response and expression of compulsive behavioral responses involving meal anticipation and consumption of large meals during situations of prolonged negative energy balance. PMID:28294152

LOX-1 in atherosclerosis: biological functions and pharmacological modifiers

PubMed Central

Xu, Suowen; Ogura, Sayoko; Chen, Jiawei; Little, Peter J.; Moss, Joel; Liu, Peiqing

2013-01-01

Lectin-like oxidized LDL (oxLDL) receptor-1 (LOX-1, also known as OLR-1), is a class E scavenger receptor that mediates the uptake of oxLDL by vascular cells. LOX-1 is involved in endothelial dysfunction, monocyte adhesion, the proliferation, migration, and apoptosis of smooth muscle cells, foam cell formation, platelet activation, as well as plaque instability; all of these events are critical in the pathogenesis of atherosclerosis. These LOX-1-dependent biological processes contribute to plaque instability and the ultimate clinical sequelae of plaque rupture and life-threatening tissue ischemia. Administration of anti-LOX-1 antibodies inhibits atherosclerosis by decreasing these cellular events. Over the past decade, multiple drugs including naturally occurring antioxidants, statins, antiinflammatory agents, antihypertensive and antihyperglycemic drugs have been demonstrated to inhibit vascular LOX-1 expression and activity. Therefore, LOX-1 represents an attractive therapeutic target for the treatment of human atherosclerotic diseases. This review aims to integrate the current understanding of LOX-1 signaling, regulation of LOX-1 by vasculoprotective drugs, and the importance of LOX-1 in the pathogenesis of atherosclerosis. PMID:23124189

Upregulation of endothelial receptor for oxidized LDL (LOX-1) by oxidized LDL and implications in apoptosis of human coronary artery endothelial cells: evidence from use of antisense LOX-1 mRNA and chemical inhibitors.

PubMed

Li, D; Mehta, J L

2000-04-01

A specific lectin-like endothelial receptor for oxidized low density lipoprotein (LOX-1), distinct from the scavenger receptor in monocytes/macrophages, has been identified and cloned. In this study, we examined the regulation of LOX-1 by oxidized low density lipoprotein (ox-LDL) and determined the role of LOX-1 in ox-LDL-induced apoptosis of cultured human coronary artery endothelial cells (HCAECs). Incubation of HCAECs with ox-LDL (40 microg/mL), but not native LDL, for 24 hours markedly increased LOX-1 expression (mRNA and protein). After 48 hours of preincubation of HCAECs with a specific antisense to LOX-1 mRNA (antisense LOX-1), ox-LDL-mediated upregulation of LOX-1 was suppressed (P<0.01). In contrast, treatment of HCAECs with sense LOX-1 had no effect. Ox-LDL also induced apoptosis (determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling and DNA laddering) of HCAECs in a concentration- and time-dependent fashion. LOX-1 played an important role in ox-LDL-mediated apoptosis of HCAECs because antisense LOX-1 inhibited this effect of ox-LDL. Polyinosinic acid and carrageenan, 2 different chemical inhibitors of LOX-1, also decreased ox-LDL-mediated apoptosis of HCAECs. Nuclear factor (NF)-kappaB was markedly activated in ox-LDL-treated HCAECs. The critical role of NF-kappaB activation became evident in experiments with antisense LOX-1, which abolished ox-LDL-mediated NF-kappaB activation. In this process, an NF-kappaB inhibitor, caffeic acid phenethyl ester, also inhibited ox-LDL-mediated apoptosis of HCAECs. These findings indicate that ox-LDL upregulates its own endothelial receptor. Ox-LDL-induced apoptosis is mediated by the action of LOX-1. In this process, NF-kappaB activation may play an important role as a signal transduction mechanism.

Evolution and Expansion of the Prokaryote-Like Lipoxygenase Family in the Brown Alga Saccharina japonica

PubMed Central

Teng, Linhong; Han, Wentao; Fan, Xiao; Xu, Dong; Zhang, Xiaowen; Dittami, Simon M.; Ye, Naihao

2017-01-01

Lipoxygenase (LOX) plays important roles in fatty acid oxidation and lipid mediator biosynthesis. In this study, we give first insights into brown algal LOX evolution. Whole genome searches revealed four, three, and eleven LOXs in Ectocarpus siliculosus, Cladosiphon okamuranus, and Saccharina japonica, respectively. In phylogenetic analyses, LOXs from brown algae form a robust clade with those from prokaryotes, suggesting an ancestral origin and slow evolution. Brown algal LOXs were divided into two clades, C1 and C2 in a phylogenetic tree. Compared to the two species of Ectocarpales, LOX gene expansion occurred in the kelp S. japonica through tandem duplication and segmental duplication. Selection pressure analysis showed that LOX genes in brown algae have undergone strong purifying selection, while the selective constraint in the C2 clade was more relaxed than that in the C1 clade. Furthermore, within each clade, LOXs of S. japonica evolved under more relaxed selection constraints than E. siliculosus and C. okamuranus. Structural modeling showed that unlike LOXs of plants and animals, which contain a β barrel in the N-terminal part of the protein, LOXs in brown algae fold into a single domain. Analysis of previously published transcriptomic data showed that LOXs in E. siliculosus are responsive to hyposaline, hypersaline, oxidative, and copper stresses. Moreover, clear divergence of expression patterns was observed among different life stages, as well as between duplicate gene pairs. In E. siliculosus, all four LOXs are male-biased in immature gametophytes, and mature gametophytes showed significantly higher LOX mRNA levels than immature gametophytes and sporophytes. In S. japonica, however, our RNA-Seq data showed that most LOXs are highly expressed in sporophytes. Even the most recently duplicated gene pairs showed divergent expression patterns, suggesting that functional divergence has likely occurred since LOX genes duplicated, which potentially contributes to the production of various oxylipins in brown algae. PMID:29234336

Inhibition of 5-LOX, COX-1, and COX-2 increases tendon healing and reduces muscle fibrosis and lipid accumulation after rotator cuff repair.

PubMed

Oak, Nikhil R; Gumucio, Jonathan P; Flood, Michael D; Saripalli, Anjali L; Davis, Max E; Harning, Julie A; Lynch, Evan B; Roche, Stuart M; Bedi, Asheesh; Mendias, Christopher L

2014-12-01

The repair and restoration of function after chronic rotator cuff tears are often complicated by muscle atrophy, fibrosis, and fatty degeneration of the diseased muscle. The inflammatory response has been implicated in the development of fatty degeneration after cuff injuries. Licofelone is a novel anti-inflammatory drug that inhibits 5-lipoxygenase (5-LOX), as well as cyclooxygenase (COX)-1 and COX-2 enzymes, which play important roles in inducing inflammation after injuries. While previous studies have demonstrated that nonsteroidal anti-inflammatory drugs and selective inhibitors of COX-2 (coxibs) may prevent the proper healing of muscles and tendons, studies about bone and cartilage have demonstrated that drugs that inhibit 5-LOX concurrently with COX-1 and COX-2 may enhance tissue regeneration. After the repair of a chronic rotator cuff tear in rats, licofelone would increase the load to failure of repaired tendons and increase the force production of muscle fibers. Controlled laboratory study. Rats underwent supraspinatus release followed by repair 28 days later. After repair, rats began a treatment regimen of either licofelone or a vehicle for 14 days, at which time animals were euthanized. Supraspinatus muscles and tendons were then subjected to contractile, mechanical, histological, and biochemical analyses. Compared with controls, licofelone-treated rats had a grossly apparent decrease in inflammation and increased fibrocartilage formation at the enthesis, along with a 62% increase in the maximum load to failure and a 51% increase in peak stress to failure. Licofelone resulted in a marked reduction in fibrosis and lipid content in supraspinatus muscles as well as reduced expression of several genes involved in fatty infiltration. Despite the decline in fibrosis and fat accumulation, muscle fiber specific force production was reduced by 23%. The postoperative treatment of cuff repair with licofelone may reduce fatty degeneration and enhance the development of a stable bone-tendon interface, although decreases in muscle fiber specific force production were observed, and force production in fact declined. This study demonstrates that the inhibition of 5-LOX, COX-1, and COX-2 modulates the healing process of repaired rotator cuff tendons. Although further studies are necessary, the treatment of patients with licofelone after cuff repair may improve the development of a stable enthesis and enhance postoperative outcomes. © 2014 The Author(s).

Human and mouse eLOX3 have distinct substrate specificities: implications for their linkage with lipoxygenases in skin

PubMed Central

Yu, Zheyong; Schneider, Claus; Boeglin, William E.; Brash, Alan R.

2008-01-01

Genetic and biochemical evidence suggests a functional link between human 12R-lipoxygenase (12R-LOX) and epidermal lipoxygenase-3 (eLOX3) in normal differentiation of the epidermis; LOX-derived fatty acid hydroperoxide is isomerized by the atypical eLOX3 into a specific epoxyalcohol that is a potential mediator in the pathway. Mouse epidermis expresses a different complement of LOX enzymes, and therefore this metabolic linkage could differ. To test this concept, we compared the substrate specificities of recombinant mouse and human eLOX3 toward sixteen hydroperoxy stereoisomers of arachidonic and linoleic acids. Both enzymes metabolized R-hydroperoxides 2–3 times faster than the corresponding S enantiomers. Whereas 12R-hydroperoxyeicosatetraenoic acid (12R-HPETE) is the best substrate for human eLOX3 (2.4 sec−1; at 30 µM substrate), mouse eLOX3 shows the highest turnover with 8R-HPETE (2.9 sec−1) followed by 8S-HPETE (1.3 sec−1). Novel product structures were characterized from reactions of mouse eLOX3 with 5S-, 8R-, and 8S-HPETEs. 8S-HPETE is converted specifically to a single epoxyalcohol, identified as 10R-hydroxy-8S,9S-epoxyeicosa-5Z,11Z,14Z-trienoic acid. The substrate preference of mouse eLOX3 and the unique occurrence of an 8S-LOX enzyme in mouse skin point to a potential LOX pathway for the production of epoxyalcohol in murine epidermal differentiation. PMID:17045234

Evaluation of serum lysyl oxidase as a blood test for colorectal cancer.

PubMed

Ward, S T; Weston, C J; Hepburn, E; Damery, S; Hejmadi, R K; Morton, D G; Middleton, G; Ismail, T; Adams, D H

2014-06-01

Lysyl oxidase (LOX) expression is elevated in colorectal cancer (CRC) tissue and associated with disease progression. A blood test may form a more acceptable diagnostic test for CRC although LOX has not previously been measured in the serum. We therefore sought to determine the clinical usefulness of a serum LOX test for CRC in a symptomatic population. Adult patients referred to a hospital colorectal clinic with bowel symptoms completed a questionnaire and provided a blood sample for serum LOX measurement. Associations between presenting symptoms, serum LOX concentrations and outcomes of investigations were tested by univariate and multivariate analyses to determine if serum LOX was clinically useful in the prediction of CRC. LOX expression in CRC and adjacent colon biopsies was evaluated by ELISA and immunohistochemistry. Thirty-one cases of colorectal cancer and 16 high-risk polyps were identified from a total of 962 participants. There was no association between serum LOX concentration and the presence of CRC, high-risk polyps or cancers at any site. LOX expression was significantly increased in CRC tissue compared to adjacent colon. Despite overexpression of LOX in CRC tissue, elevated serum levels could not be demonstrated. Serum LOX measurement is therefore not a clinically useful test for CRC. Copyright © 2013 Elsevier Ltd. All rights reserved.

Structure and Ligand Based Drug Design Strategies in the Development of Novel 5-LOX Inhibitors

PubMed Central

Aparoy, Polamarasetty; Kumar Reddy, Kakularam; Reddanna, Pallu

2012-01-01

Lipoxygenases (LOXs) are non-heme iron containing dioxygenases involved in the oxygenation of polyunsaturated fatty acids (PUFAs) such as arachidonic acid (AA). Depending on the position of insertion of oxygen, LOXs are classified into 5-, 8-, 9-, 12- and 15-LOX. Among these, 5-LOX is the most predominant isoform associated with the formation of 5-hydroperoxyeicosatetraenoic acid (5-HpETE), the precursor of non-peptido (LTB4) and peptido (LTC4, LTD4, and LTE4) leukotrienes. LTs are involved in inflammatory and allergic diseases like asthma, ulcerative colitis, rhinitis and also in cancer. Consequently 5-LOX has become target for the development of therapeutic molecules for treatment of various inflammatory disorders. Zileuton is one such inhibitor of 5-LOX approved for the treatment of asthma. In the recent times, computer aided drug design (CADD) strategies have been applied successfully in drug development processes. A comprehensive review on structure based drug design strategies in the development of novel 5-LOX inhibitors is presented in this article. Since the crystal structure of 5-LOX has been recently solved, efforts to develop 5-LOX inhibitors have mostly relied on ligand based rational approaches. The present review provides a comprehensive survey on these strategies in the development of 5-LOX inhibitors. PMID:22680930

Coil-On-Plug Ignition for LOX/Methane Liquid Rocket Engines in Thermal Vacuum Environments

NASA Technical Reports Server (NTRS)

Melcher, John C.; Atwell, Matthew J.; Morehead, Robert L.; Hurlbert, Eric A.; Bugarin, Luz; Chaidez, Mariana

2017-01-01

A coil-on-plug ignition system has been developed and tested for Liquid Oxygen (LOX) / liquid methane rocket engines operating in thermal vacuum conditions. The igniters were developed and tested as part of the Integrated Cryogenic Propulsion Test Article (ICPTA), previously tested as part of the Project Morpheus test vehicle. The ICPTA uses an integrated, pressure-fed, cryogenic LOX/methane propulsion system including a reaction control system (RCS) and a main engine. The ICPTA was tested at NASA Glenn Research Center's Plum Brook Station in the Spacecraft Propulsion Research Facility (B-2) under vacuum and thermal vacuum conditions. In order to successfully demonstrate ignition reliability in the vacuum conditions and eliminate corona discharge issues, a coil-on-plug ignition system has been developed. The ICPTA uses spark-plug ignition for both the main engine igniter and the RCS. The coil-on-plug configuration eliminates the conventional high-voltage spark plug cable by combining the coil and the spark-plug into a single component. Prior to ICPTA testing at Plum Brook, component-level reaction control engine (RCE) and main engine igniter testing was conducted at NASA Johnson Space Center (JSC), which demonstrated successful hot-fire ignition using the coil-on-plug from sea-level ambient conditions down to 10(exp.-2) torr. Integrated vehicle hot-fire testing at JSC demonstrated electrical and command/data system performance. Lastly, Plum Brook testing demonstrated successful ignitions at simulated altitude conditions at 30 torr and cold thermal-vacuum conditions at 6 torr. The test campaign successfully proved that coil-on-plug technology will enable integrated LOX/methane propulsion systems in future spacecraft.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Bin; Liu, Ning-Ning; Liu, Wei-Hua

Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) and GATA Binding Protein 4 (GATA4) are important for the growth of cardiac fibroblasts (CFs). When deregulated, LOX-1 and GATA4 can cause cardiac remodeling. In the present study, we found novel evidence that GATA4 was required for the LOX-1 regulation of CF proliferation. The inhibition of LOX-1 by RNA interference LOX-1 lentivirus resulted in the loss of PI3K/Akt activation and GATA4 protein expression. The overexpression of LOX-1 by lentivirus rescued CF proliferation, PI3K/Akt activation, and GATA4 protein expression. Moreover, GATA4 overexpression enhanced CF proliferation with LOX-1 inhibition. We also found that the inhibition ofmore » PI3K/Akt activation by LY294002, a PI3K inhibitor, reduced cell proliferation and protein level of GATA4. In summary, GATA4 may play an important role in the LOX-1 and PI3K/Akt regulation of CF proliferation. -- Highlights: •GATA4 is regulated by LOX-1 signaling in CFs. •GATA4 is involved in LOX-1 regulating CF proliferation. •GATA4 is regulated by PI3K/Akt signaling in CFs.« less

Cloning and characterization of a 9-lipoxygenase gene induced by pathogen attack from Nicotiana benthamiana for biotechnological application

PubMed Central

2011-01-01

Background Plant lipoxygenases (LOXs) have been proposed to form biologically active compounds both during normal developmental stages such as germination or growth as well as during responses to environmental stress such as wounding or pathogen attack. In our previous study, we found that enzyme activity of endogenous 9-LOX in Nicotiana benthamiana was highly induced by agroinfiltration using a tobacco mosaic virus (TMV) based vector system. Results A LOX gene which is expressed after treatment of the viral vectors was isolated from Nicotiana benthamiana. As the encoded LOX has a high amino acid identity to other 9-LOX proteins, the gene was named as Nb-9-LOX. It was heterologously expressed in yeast cells and its enzymatic activity was characterized. The yeast cells expressed large quantities of stable 9-LOX (0.9 U ml-1 cell cultures) which can oxygenate linoleic acid resulting in high yields (18 μmol ml-1 cell cultures) of hydroperoxy fatty acid. The product specificity of Nb-9-LOX was examined by incubation of linoleic acid and Nb-9-LOX in combination with a 13-hydroperoxide lyase from watermelon (Cl-13-HPL) or a 9/13-hydroperoxide lyase from melon (Cm-9/13-HPL) and by LC-MS analysis. The result showed that Nb-9-LOX possesses both 9- and 13-LOX specificity, with high predominance for the 9-LOX function. The combination of recombinant Nb-9-LOX and recombinant Cm-9/13-HPL produced large amounts of C9-aldehydes (3.3 μmol mg-1 crude protein). The yield of C9-aldehydes from linoleic acid was 64%. Conclusion The yeast expressed Nb-9-LOX can be used to produce C9-aldehydes on a large scale in combination with a HPL gene with 9-HPL function, or to effectively produce 9-hydroxy-10(E),12(Z)-octadecadienoic acid in a biocatalytic process in combination with cysteine as a mild reducing agent. PMID:21450085

Stereocontrol of Arachidonic Acid Oxygenation by Vertebrate Lipoxygenases

PubMed Central

Jansen, Christian; Hofheinz, Katharina; Vogel, Robert; Roffeis, Jana; Anton, Monika; Reddanna, Pallu; Kuhn, Hartmut; Walther, Matthias

2011-01-01

Animal lipoxygenases (LOXs) are classified according to their specificity of arachidonic acid oxygenation, and previous sequence alignments suggested that S-LOXs contain a conserved Ala at a critical position at the active site but R-LOXs carry a Gly instead. Here we cloned, expressed, and characterized a novel LOX isoform from the model vertebrate Danio rerio (zebrafish) that carries a Gly at this critical position, classifying this enzyme as putative arachidonic acid R-LOX. Surprisingly, the almost exclusive arachidonic acid oxygenation product was 12S-H(p)ETE (hydro(pero)xyeicosatetraenoic acid), and extensive mutation around Gly-410 failed to induce R-lipoxygenation. This finding prompted us to explore the importance of the corresponding amino acids in other vertebrate S-LOXs. We found that Ala-to-Gly exchange in human 15-LOX2 and human platelet 12-LOX induced major alterations in the reaction specificity with an increase of specific R-oxygenation products. For mouse 5-LOX and 12/15-LOX from rabbits, men, rhesus monkeys, orangutans, and mice, only minor alterations in the reaction specificity were observed. For these enzymes, S-HETE (hydroxyeicosatetraenoic acid) isomers remained the major oxygenation products, whereas chiral R-HETEs contributed only 10–30% to the total product mixture. Taken together these data indicate that the Ala-versus-Gly concept may not always predict the reaction specificity of vertebrate LOX isoforms. PMID:21880725

OsLOX2, a rice type I lipoxygenase, confers opposite effects on seed germination and longevity.

PubMed

Huang, Jiexue; Cai, Maohong; Long, Qizhang; Liu, Linglong; Lin, Qiuyun; Jiang, Ling; Chen, Saihua; Wan, Jianmin

2014-08-01

Rice production and seed storage are confronted with grain deterioration and loss of seed viability. Some members of the lipoxygenase (LOX) family function in degradation of storage lipids during the seed germination, but little is known about their influence on seed longevity during storage. We characterized the role of rice OsLOX2 gene in seed germination and longevity via over-expression and knock-down approaches. Abundant expression of OsLOX2 was detected in panicles, roots, and stems, but not in leaves. Moreover, OsLOX2 was highly induced during germination. OsLOX2 protein, located in the cytoplasm, showed a wide range of temperature adaptation (20-50 °C) and a substrate preference to linoleic acid. Lines over-expressing OsLOX2 showed accelerated seed germination under normal condition and lower seed viability after accelerated aging. RNA interference (RNAi) of OsLOX2 caused delayed germination and enhanced seed longevity. RNAi lines with strongly repressed OsLOX2 activity completely lost the capability of germination after accelerated aging. More lipid hydroperoxide were found in OE15 than the control, but less in RNAi lines than in the WT Nipponbare. Therefore, OsLOX2 acts in opposite directions during seed germination and longevity during storage. Appropriate repression of the OsLOX2 gene may delay the aging process during the storage without compromising germination under normal conditions.

The Persimmon 9-lipoxygenase Gene DkLOX3 Plays Positive Roles in Both Promoting Senescence and Enhancing Tolerance to Abiotic Stress

PubMed Central

Hou, Yali; Meng, Kun; Han, Ye; Ban, Qiuyan; Wang, Biao; Suo, Jiangtao; Lv, Jingyi; Rao, Jingping

2015-01-01

The lipoxygenase (LOX) pathway is a key regulator for lipid peroxidation, which is crucial for plant senescence and defense pathways. In this study, the transcriptional expression patterns of three persimmon (Diospyros kaki L. ‘Fupingjianshi’) 9-lipoxygenase genes (DkLOX1, DkLOX3, and DkLOX4) were investigated. DkLOX1 was specifically expressed in fruit, particularly in young fruit, and showed little response to the postharvest environments. DkLOX4 was expressed in all tissues and slightly stimulated by mechanical damage and low temperature. DkLOX3 was expressed mainly in mature fruit, and the expression was extremely high throughout the storage period, apparently up-regulated by mechanical damage and high carbon dioxide treatments. Further functional analysis showed that overexpression of DkLOX3 in tomato (Solanum lycopersicum cv. Micro-Tom) accelerated fruit ripening and softening. This was accompanied by higher malondialdehyde (MDA) content and lycopene accumulation, advanced ethylene release peak and elevated expression of ethylene synthesis genes, including ACS2, ACO1, and ACO3. In addition, DkLOX3 overexpression promoted dark induced transgenic Arabidopsis leaf senescence with more chlorophyll loss, increased electrolyte leakage and MDA content. Furthermore, the functions of DkLOX3 in response to abiotic stresses, including osmotic stress, high salinity and drought were investigated. Arabidopsis DkLOX3 overexpression (DkLOX3-OX) transgenic lines were found to be more tolerant to osmotic stress with higher germination rate and root growth than wild-type. Moreover, DkLOX3-OX Arabidopsis plants also exhibited enhanced resistance to high salinity and drought, with similar decreased O2- and H2O2 accumulation and upregulation of stress-responsive genes expression, including RD22, RD29A, RD29B, and NCED3, except for FRY1, which plays a negative role in stress response. Overall, these results suggested that DkLOX3 plays positive roles both in promoting ripening and senescence through lipid peroxidation and accelerated ethylene production and in stress response via regulating reactive oxygen species accumulation and stress responsive genes expression. PMID:26697033

Nonlinear effects in a plain journal bearing. I - Analytical study. II - Results

NASA Technical Reports Server (NTRS)

Choy, F. K.; Braun, M. J.; Hu, Y.

1991-01-01

In the first part of this work, a numerical model is presented which couples the variable-property Reynolds equation with a rotor-dynamics model for the calculation of a plain journal bearing's nonlinear characteristics when working with a cryogenic fluid, LOX. The effects of load on the linear/nonlinear plain journal bearing characteristics are analyzed and presented in a parametric form. The second part of this work presents numerical results obtained for specific parametric-study input variables (lubricant inlet temperature, external load, angular rotational speed, and axial misalignment). Attention is given to the interrelations between pressure profiles and bearing linear and nonlinear characteristics.

Conceptual Design, Feasibility and Payoff Analysis of a Third Stage for EELV (Briefing Charts)

DTIC Science & Technology

2014-07-30

performance capability – Hydrazine, N2O4/MMH, LOX/RP, H2O2/RP, LOX/CH4, LOX/ LH2 (MR of 6), and LOX/ LH2 (MR of 10) • Propulsion system split into multiple...area ratio attainable within length envelope – Use of four chambers allows area ratios >150 – LOX/ LH2 (MR=6) is highest performing propellant...combination •Due to LOX/ LH2 (MR=6) low bulk density it is not possible to store as much propellant within stage – Resulting in the lowest total

Stoichiometry of the Cre recombinase bound to the lox recombining site.

PubMed Central

Mack, A; Sauer, B; Abremski, K; Hoess, R

1992-01-01

The site-specific recombinase Cre from bacteriophage P1 binds and carries out recombination at a 34 bp lox site. The lox site consists of two 13 bp inverted repeats, separated by an 8 bp spacer region. Both the palindromic nature of the site and the results of footprinting and band shift experiments suggest that a minimum of two Cre molecules bind to a lox site. We report here experiments that demonstrate the absolute stoichiometry of the Cre-lox complex to be one molecule of Cre bound per inverted repeat, or two molecules per lox site. Images PMID:1408747

Lipoxygenase is involved in the control of potato tuber development.

PubMed

Kolomiets, M V; Hannapel, D J; Chen, H; Tymeson, M; Gladon, R J

2001-03-01

Plant lipoxygenases (LOXs) are a functionally diverse class of dioxygenases implicated in physiological processes such as growth, senescence, and stress-related responses. LOXs incorporate oxygen into their fatty acid substrates and produce hydroperoxide fatty acids that are precursors of jasmonic acid and related compounds. Here, we report the involvement of the tuber-associated LOXs, designated the Lox1 class, in the control of tuber growth. RNA hybridization analysis showed that the accumulation of Lox1 class transcripts was restricted to developing tubers, stolons, and roots and that mRNA accumulation correlated positively with tuber initiation and growth. In situ hybridization showed that Lox1 class transcripts accumulated in the apical and subapical regions of the newly formed tuber, specifically in the vascular tissue of the perimedullary region, the site of the most active cell growth during tuber enlargement. Suppression mutants produced by expressing antisense coding sequence of a specific tuber LOX, designated POTLX-1, exhibited a significant reduction in LOX activity in stolons and tubers. The suppression of LOX activity correlated with reduced tuber yield, decreased average tuber size, and a disruption of tuber formation. Our results indicate that the pathway initiated by the expression of the Lox1 class genes of potato is involved in the regulation of tuber enlargement.

Identification of a unique nsaid, fluoro-loxoprofen with gastroprotective activity.

PubMed

Suemasu, Shintaro; Yamakawa, Naoki; Ishihara, Tomoaki; Asano, Teita; Tahara, Kayoko; Tanaka, Ken-ichiro; Matsui, Hirofumi; Okamoto, Yoshinari; Otsuka, Masami; Takeuchi, Koji; Suzuki, Hidekazu; Mizushima, Tohru

2012-12-01

We previously proposed that direct cytotoxicity of NSAIDs due to their membrane permeabilization activity, together with their ability to decrease gastric prostaglandin E(2), contributes to production of gastric lesions. Compared to loxoprofen (LOX), fluoro-loxoprofen (F-LOX) has much lower membrane permeabilization and gastric ulcerogenic activities but similar anti-inflammatory activity. In this study, we examined the mechanism for this low ulcerogenic activity in rats. Compared to LOX, the level of gastric mucosal cell death was lower following administration of F-LOX. However, the gastric level of prostaglandin E(2) was similar in response to treatment with the two NSAIDs. Oral pre-administration of F-LOX conferred protection against the formation of gastric lesions produced by subsequent administration of LOX and orally administered F-LOX resulted in a higher gastric pH value and mucus content. In the presence of a stimulant of gastric acid secretion, the difference in the ulcerogenic activity of F-LOX and LOX was less apparent. Furthermore, an increase in the mucus was observed in gastric cells cultured in the presence of F-LOX in a manner dependent of increase in the cellular level of cAMP. These results suggest that low ulcerogenic activity of F-LOX involves its both low direct cytotoxicity and protective effect against the development of gastric lesions. This protective effect seems to be mediated through an increase in a protective factor (mucus) and a decrease in an aggressive factor (acid). Copyright © 2012 Elsevier Inc. All rights reserved.

Lysyl Oxidase Induces Vascular Oxidative Stress and Contributes to Arterial Stiffness and Abnormal Elastin Structure in Hypertension: Role of p38MAPK.

PubMed

Martínez-Revelles, Sonia; García-Redondo, Ana B; Avendaño, María S; Varona, Saray; Palao, Teresa; Orriols, Mar; Roque, Fernanda R; Fortuño, Ana; Touyz, Rhian M; Martínez-González, Jose; Salaices, Mercedes; Rodríguez, Cristina; Briones, Ana M

2017-09-01

Vascular stiffness, structural elastin abnormalities, and increased oxidative stress are hallmarks of hypertension. Lysyl oxidase (LOX) is an elastin crosslinking enzyme that produces H 2 O 2 as a by-product. We addressed the interplay between LOX, oxidative stress, vessel stiffness, and elastin. Angiotensin II (Ang II)-infused hypertensive mice and spontaneously hypertensive rats (SHR) showed increased vascular LOX expression and stiffness and an abnormal elastin structure. Mice over-expressing LOX in vascular smooth muscle cells (TgLOX) exhibited similar mechanical and elastin alterations to those of hypertensive models. LOX inhibition with β-aminopropionitrile (BAPN) attenuated mechanical and elastin alterations in TgLOX mice, Ang II-infused mice, and SHR. Arteries from TgLOX mice, Ang II-infused mice, and/or SHR exhibited increased vascular H 2 O 2 and O 2 .- levels, NADPH oxidase activity, and/or mitochondrial dysfunction. BAPN prevented the higher oxidative stress in hypertensive models. Treatment of TgLOX and Ang II-infused mice and SHR with the mitochondrial-targeted superoxide dismutase mimetic mito-TEMPO, the antioxidant apocynin, or the H 2 O 2 scavenger polyethylene glycol-conjugated catalase (PEG-catalase) reduced oxidative stress, vascular stiffness, and elastin alterations. Vascular p38 mitogen-activated protein kinase (p38MAPK) activation was increased in Ang II-infused and TgLOX mice and this effect was prevented by BAPN, mito-TEMPO, or PEG-catalase. SB203580, the p38MAPK inhibitor, normalized vessel stiffness and elastin structure in TgLOX mice. We identify LOX as a novel source of vascular reactive oxygen species and a new pathway involved in vascular stiffness and elastin remodeling in hypertension. LOX up-regulation is associated with enhanced oxidative stress that promotes p38MAPK activation, elastin structural alterations, and vascular stiffness. This pathway contributes to vascular abnormalities in hypertension. Antioxid. Redox Signal. 27, 379-397.

Lysyl Oxidase Induces Vascular Oxidative Stress and Contributes to Arterial Stiffness and Abnormal Elastin Structure in Hypertension: Role of p38MAPK

PubMed Central

Martínez-Revelles, Sonia; García-Redondo, Ana B.; Avendaño, María S.; Varona, Saray; Palao, Teresa; Orriols, Mar; Roque, Fernanda R.; Fortuño, Ana; Touyz, Rhian M.; Martínez-González, Jose; Salaices, Mercedes

2017-01-01

Abstract Aims: Vascular stiffness, structural elastin abnormalities, and increased oxidative stress are hallmarks of hypertension. Lysyl oxidase (LOX) is an elastin crosslinking enzyme that produces H2O2 as a by-product. We addressed the interplay between LOX, oxidative stress, vessel stiffness, and elastin. Results: Angiotensin II (Ang II)-infused hypertensive mice and spontaneously hypertensive rats (SHR) showed increased vascular LOX expression and stiffness and an abnormal elastin structure. Mice over-expressing LOX in vascular smooth muscle cells (TgLOX) exhibited similar mechanical and elastin alterations to those of hypertensive models. LOX inhibition with β-aminopropionitrile (BAPN) attenuated mechanical and elastin alterations in TgLOX mice, Ang II-infused mice, and SHR. Arteries from TgLOX mice, Ang II-infused mice, and/or SHR exhibited increased vascular H2O2 and O2.− levels, NADPH oxidase activity, and/or mitochondrial dysfunction. BAPN prevented the higher oxidative stress in hypertensive models. Treatment of TgLOX and Ang II-infused mice and SHR with the mitochondrial-targeted superoxide dismutase mimetic mito-TEMPO, the antioxidant apocynin, or the H2O2 scavenger polyethylene glycol-conjugated catalase (PEG-catalase) reduced oxidative stress, vascular stiffness, and elastin alterations. Vascular p38 mitogen-activated protein kinase (p38MAPK) activation was increased in Ang II-infused and TgLOX mice and this effect was prevented by BAPN, mito-TEMPO, or PEG-catalase. SB203580, the p38MAPK inhibitor, normalized vessel stiffness and elastin structure in TgLOX mice. Innovation: We identify LOX as a novel source of vascular reactive oxygen species and a new pathway involved in vascular stiffness and elastin remodeling in hypertension. Conclusion: LOX up-regulation is associated with enhanced oxidative stress that promotes p38MAPK activation, elastin structural alterations, and vascular stiffness. This pathway contributes to vascular abnormalities in hypertension. Antioxid. Redox Signal. 27, 379–397. PMID:28010122

Loxoprofen sodium suppresses mouse tumor growth by inhibiting vascular endothelial growth factor.

PubMed

Kanda, Akio; Ebihara, Satoru; Takahashi, Hidenori; Sasaki, Hidetada

2003-01-01

There is increasing evidence to suggest the anti-tumor effects of non-steroidal anti-inflammatory drugs (NSAIDs). In this study it was shown that the most popular NSAID in Japan, loxoprofen sodium (LOX), inhibited in vivo growth of implanted Lewis lung carcinoma (LLC), whereas LOX did not affect the proliferation and viability of LLC cells in vitro. Intratumoral vessel density in LOX-treated mice was significantly lower than that of mice without treatment. Intratumoral expressions of vascular endothelial growth factor (VEGF) mRNA were attenuated by the LOX treatment. LOX suppressed both intratumoral and systemic VEGF protein in LLC-implanted mice. LOX also inhibited tubular formation of primary cultured human umbilical vein endothelial cells, presumably due to the inhibition of VEGF. In patients with advanced non-small cell lung cancer, LOX medication (120 mg/day) for a week significantly decreased the plasma VEGF level. These results suggest that LOX may have potent anti-cancer effects in patients with advanced NSCLC.

A Lipoxygenase from Red Alga Pyropia haitanensis, a Unique Enzyme Catalyzing the Free Radical Reactions of Polyunsaturated Fatty Acids with Triple Ethylenic Bonds

PubMed Central

Zhu, Zhujun; Qian, Feijian; Yang, Rui; Chen, Juanjuan; Luo, Qijun; Chen, Haimin; Yan, Xiaojun

2015-01-01

Lipoxygenases (LOXs) are key enzymes to regulate the production of hormones and defensive metabolites in plants, animals and algae. In this research, a full length LOX gene has been cloned and expressed from the red alga Pyropia haitanensis (Bangiales, Rhodophyta) gametophyte (PhLOX2). Subsequent phylogenetic analysis showed that such LOX enzymes are separated at the early stage of evolution, establishing an independent branch. The LOX activity was investigated at the optimal pH of 8.0. It appears that PhLOX2 is a multifunctional enzyme featuring both lipoxygenase and hydroperoxidase activities. Additionally, PhLOX2 exhibits remarkable substrate and position flexibility, and it can catalyze an array of chemical reactions involving various polyunsaturated fatty acids, ranging from C18 to C22. As a matter of fact, mono-hydroperoxy, di-hydroperoxy and hydroxyl products have been obtained from such transformations, and eicosapentaenoic acid seem to be the most preferred substrate. It was found that at least triple ethylenic bonds are required for PhLOX2 to function as a LOX, and the resulting hydroxy products should be originated from the PhLOX2 mediated reduction of mono-hydroperoxides, in which the hydrogen abstraction occurs on the carbon atom between the second and third double bond. Most of the di-hydroperoxides observed seem to be missing their mono-position precursors. The substrate and position flexibility, as well as the function versatility of PhLOXs represent the ancient enzymatic pathway for organisms to control intracellular oxylipins. PMID:25658744

Tripropellant engine study

NASA Technical Reports Server (NTRS)

Wheeler, D. B.; Kirby, F. M.

1978-01-01

The potential for converting the space shuttle main engine (SSME) to a dual-fuel, dual-mode engine using LOX/hydrocarbon propellants in mode 1 and LOX/H2 in mode 2 was examined. Various engine system concepts were formulated that included staged combustion and gas generator turbine power cycles, and LOX/RP-1, LOX/CH4, and LOX/C3H8 mode 1 propellants. Both oxidizer and fuel regenerative cooling were considered. All of the SSME major components were examined to determine their adaptability to the candidate dual-fuel engines.

Lipoxygenase activity in different species of sweet lupin (Lupinus L.) seeds and flakes.

PubMed

Stephany, Michael; Bader-Mittermaier, Stephanie; Schweiggert-Weisz, Ute; Carle, Reinhold

2015-05-01

Lipoxygenase (LOX)-catalysed degradation of polyunsaturated fatty acids is supposed to be a major cause of undesirable off-flavour development in legumes. In the present study, a photometric LOX assay including adequate sample workup was adapted to lupin seeds, kernels and flakes, respectively. Optimum reaction conditions were at pH 7.5 using a phosphate buffer concentration of 150 mmol l(-1) without the addition of sodium chloride. The LOX activities of different lupin species and varieties were compared. Significant variations among the species and varieties ranging from 50 to 1004 units mg(-1) protein were determined, being significantly lower than soybean LOX activity. Hulling and flaking of the seeds resulted in a 15% increase of LOX activity. In contrast to soy and other legumes, LOX from lupin only converted free fatty acids, whereas trilinolein and β-carotene were not oxidised. Consequently, according to the established classification, lupin LOX activity may be assigned to the LOX type-1, which, to the best of our knowledge, was demonstrated for the first time. Copyright © 2014 Elsevier Ltd. All rights reserved.

Cre recombinase-mediated site-specific recombination between plant chromosomes.

PubMed Central

Qin, M; Bayley, C; Stockton, T; Ow, D W

1994-01-01

We report the use of the bacteriophage P1 Cre-lox system for generating conservative site-specific recombination between tobacco chromosomes. Two constructs, one containing a promoterless hygromycin-resistance gene preceded by a lox site (lox-hpt) and the other containing a cauliflower mosaic virus 35S promoter linked to a lox sequence and the cre coding region (35S-lox-cre), were introduced separately into tobacco plants. Crosses between plants harboring either construct produced plants with the two constructs situated on different chromosomes. Plants with recombination events were identified by selecting for hygromycin resistance, a phenotype expressed upon recombination. Molecular analysis showed that these recombination events occurred specifically at the lox sites and resulted in the reciprocal exchange of flanking host DNA. Progenies of these plants showed 67-100% cotransmission of the new transgenes, 35S-lox-hpt and lox-cre, consistent with the preferential cosegregation of translocated chromosomes. These results illustrate that site-specific recombination systems can be useful tools for the large-scale manipulation of eukaryotic chromosomes in vivo. Images PMID:8127869

Root-Derived Oxylipins Promote Green Peach Aphid Performance on Arabidopsis Foliage[W

PubMed Central

Nalam, Vamsi J.; Keeretaweep, Jantana; Sarowar, Sujon; Shah, Jyoti

2012-01-01

Oxylipins function as signaling molecules in plant growth and development and contribute to defense against stress. Here, we show that oxylipins also facilitate infestation of Arabidopsis thaliana shoots by the phloem sap–consuming green peach aphid (GPA; Myzus persicae), an agronomically important insect pest. GPAs had difficulty feeding from sieve elements and tapping into the xylem of lipoxygenase5 (lox5) mutant plants defective in LOX activity. These defects in GPA performance in the lox5 mutant were accompanied by reduced water content of GPAs and a smaller population size of GPAs in the mutant compared with the wild-type plant. LOX5 expression was rapidly induced in roots in response to infestation of shoots by GPAs. In parallel, levels of LOX5-derived oxylipins increased in roots and in petiole exudates of GPA-colonized plants. Application of 9-hydroxyoctadecadienoic acid (an oxylipin produced by the LOX5 enzyme) to roots restored water content and GPA population size in lox5 plants, thus confirming that a LOX5-derived oxylipin promotes infestation of the foliage by GPAs. Micrografting experiments demonstrated that GPA performance on foliage is influenced by the LOX5 genotype in roots, thus demonstrating the importance of root-derived oxylipins in colonization of aboveground organs by an insect. PMID:22474183

Comparative analysis of lysyl oxidase (like) family members in pulmonary fibrosis.

PubMed

Aumiller, Verena; Strobel, Benjamin; Romeike, Merrit; Schuler, Michael; Stierstorfer, Birgit E; Kreuz, Sebastian

2017-03-10

Extracellular matrix (ECM) composition and stiffness are major driving forces for the development and persistence of fibrotic diseases. Lysyl oxidase (LOX) and LOX-like (LOXL) proteins play crucial roles in ECM remodeling due to their collagen crosslinking and intracellular functions. Here, we systematically investigated LOX/L expression in primary fibroblasts and epithelial cells under fibrotic conditions, Bleomycin (BLM) induced lung fibrosis and in human IPF tissue. Basal expression of all LOX/L family members was detected in epithelial cells and at higher levels in fibroblasts. Various pro-fibrotic stimuli broadly induced LOX/L expression in fibroblasts, whereas specific induction of LOXL2 and partially LOX was observed in epithelial cells. Immunohistochemical analysis of lung tissue from 14 IPF patients and healthy donors revealed strong induction of LOX and LOXL2 in bronchial and alveolar epithelium as well as fibroblastic foci. Using siRNA experiments we observed that LOXL2 and LOXL3 were crucial for fibroblast-to-myofibroblast transition (FMT). As FMT could only be reconstituted with an enzymatically active LOXL2 variant, we conclude that LOXL2 enzymatic function is crucial for fibroblast transdifferentiation. In summary, our study provides a comprehensive analysis of the LOX/L family in fibrotic lung disease and indicates prominent roles for LOXL2/3 in fibroblast activation and LOX/LOXL2 in IPF.

LOX-1, OxLDL, and Atherosclerosis

PubMed Central

Catapano, Alberico Luigi

2013-01-01

Oxidized low-density lipoprotein (OxLDL) contributes to the atherosclerotic plaque formation and progression by several mechanisms, including the induction of endothelial cell activation and dysfunction, macrophage foam cell formation, and smooth muscle cell migration and proliferation. Vascular wall cells express on their surface several scavenger receptors that mediate the cellular effects of OxLDL. The lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is the main OxLDL receptor of endothelial cells, and it is expressed also in macrophages and smooth muscle cells. LOX-1 is almost undetectable under physiological conditions, but it is upregulated following the exposure to several proinflammatory and proatherogenic stimuli and can be detected in animal and human atherosclerotic lesions. The key contribution of LOX-1 to the atherogenic process has been confirmed in animal models; LOX-1 knockout mice exhibit reduced intima thickness and inflammation and increased expression of protective factors; on the contrary, LOX-1 overexpressing mice present an accelerated atherosclerotic lesion formation which is associated with increased inflammation. In humans, LOX-1 gene polymorphisms were associated with increased susceptibility to myocardial infarction. Inhibition of the LOX-1 receptor with chemicals or antisense nucleotides is currently being investigated and represents an emerging approach for controlling OxLDL-LOX-1 mediated proatherogenic effects. PMID:23935243

Conditional Deletion of the Pten Gene in the Mouse Prostate Induces Prostatic Intraepithelial Neoplasms at Early Ages but a Slow Progression to Prostate Tumors

PubMed Central

Zhu, Chunfang; Lee, Suk Hyung; Ye, Ding-Wei; Luong, Richard; Sun, Zijie

2013-01-01

The PTEN tumor suppressor gene is frequently inactivated in human prostate cancer. Using Osr1 (odd skipped related 1)-Cre mice, we generated a novel conditional Pten knockout mouse strain, PtenLoxP:Osr1-Cre. Conditional biallelic and monoallelic Pten knockout mice were viable. Deletion of Pten expression was detected in the prostate of PtenLoxP/LoxP:Osr1-Cre mice as early as 2 weeks of age. Intriguingly, PtenLoxP/LoxP:Osr1-Cre mice develop high-grade prostatic intraepithelial neoplasms (PINs) with high penetrance as early as one-month of age, and locally invasive prostatic tumors after 12-months of age. PtenLoxP/+:Osr1-Cre mice show only mild oncogenic changes after 8-weeks of age. Castration of PtenLoxP/LoxP:Osr1-Cre mice shows no significant regression of prostate tumors, although a shift of androgen receptor (AR) staining from the nuclei to cytoplasm is observed in Pten null tumor cells of castrated mice. Enhanced Akt activity is observed in Pten null tumor cells of castrated PtenLoxP/LoxP:Osr1-Cre. This study provides a novel mouse model that can be used to investigate a primary role of Pten in initiating oncogenic transformation in the prostate and to examine other genetic and epigenetic changes that are required for tumor progression in the mouse prostate. PMID:23308230

Investigation of calcium-dependent activity and conformational dynamics of zebra fish 12-lipoxygenase.

PubMed

Mittal, Monica; Hasan, Mahmudul; Balagunaseelan, Navisraj; Fauland, Alexander; Wheelock, Craig; Rådmark, Olof; Haeggström, Jesper Z; Rinaldo-Matthis, Agnes

2017-08-01

A 12-lipoxygenase in zebra fish (zf12-LOX) was found to be required for normal embryonic development and LOXs are of great interest for targeted drug designing. In this study, we investigate the structural-functional aspects of zf12-LOX in response to calcium. A soluble version of zf12-LOX was created by mutagenesis. Based on multiple sequence alignment, we mutated the putative calcium-responsive amino acids in N-PLAT domain of soluble zf12-LOX. Using a series of biophysical methods, we ascertained the oligomeric state, stability, structural integrity and conformational changes of zf12-LOX in response to calcium. We also compared the biophysical properties of soluble zf12-LOX with the mutant in the absence and presence of calcium. Here we provide a detailed characterization of soluble zf12-LOX and the mutant. Both proteins exist as compact monomers in solution, however the enzyme activity of soluble zf12-LOX is significantly increased in presence of calcium. We find that the stimulatory effect of calcium on zf12-LOX is related to a change in protein structure as observed by SAXS, adopting an open-state. In contrast, enzyme with a mutated calcium regulatory site has reduced activity-response to calcium and restricted large re-modeling, suggesting that it retains a closed-state in response to calcium. Taken together, our study suggests that Ca 2+ -dependent regulation is associated with different domain conformation(s) that might change the accessibility to substrate-binding site in response to calcium. The study can be broadly implicated in better understanding the mode(s) of action of LOXs, and the enzymes regulated by calcium in general. Copyright © 2017 Elsevier B.V. All rights reserved.

The elicitation of a systemic resistance by Pseudomonas putida BTP1 in tomato involves the stimulation of two lipoxygenase isoforms

PubMed Central

2011-01-01

Background Some non-pathogenic rhizobacteria called Plant Growth Promoting Rhizobacteria (PGPR) possess the capacity to induce in plant defense mechanisms effective against pathogens. Precedent studies showed the ability of Pseudomonas putida BTP1 to induce PGPR-mediated resistance, termed ISR (Induced Systemic Resistance), in different plant species. Despite extensive works, molecular defense mechanisms involved in ISR are less well understood that in the case of pathogen induced systemic acquired resistance. Results We analyzed the activities of phenylalanine ammonia-lyase (PAL) and lipoxygenase (LOX), key enzymes of the phenylpropanoid and oxylipin pathways respectively, in tomato treated or not with P. putida BTP1. The bacterial treatment did not stimulate PAL activity and linoleate-consuming LOX activities. Linolenate-consuming LOX activity, on the contrary, was significantly stimulated in P. putida BTP1-inoculated plants before and two days after infection by B. cinerea. This stimulation is due to the increase of transcription level of two isoforms of LOX: TomLoxD and TomLoxF, a newly identified LOX gene. We showed that recombinant TomLOXF preferentially consumes linolenic acid and produces 13-derivative of fatty acids. After challenging with B. cinerea, the increase of transcription of these two LOX genes and higher linolenic acid-consuming LOX activity were associated with a more rapid accumulation of free 13-hydroperoxy-octadecatrienoic and 13-hydroxy-octadecatrienoic acids, two antifungal oxylipins, in bacterized plants. Conclusion In addition to the discovery of a new LOX gene in tomato, this work is the first to show differential induction of LOX isozymes and a more rapid accumulation of 13-hydroperoxy-octadecatrienoic and 13-hydroxy-octadecatrienoic acids in rhizobacteria mediated-induced systemic resistance. PMID:21294872

The lipoxygenase gene ALOXE3 implicated in skin differentiation encodes a hydroperoxide isomerase

PubMed Central

Yu, Zheyong; Schneider, Claus; Boeglin, William E.; Marnett, Lawrence J.; Brash, Alan R.

2003-01-01

Lipoxygenase (LOX) enzymes form fatty acid hydroperoxides used in membrane remodeling and cell signaling. Mammalian epidermal LOX type 3 (eLOX3) is distinctive in totally lacking this typical oxygenase activity. Surprisingly, genetic evidence has linked mutations in eLOX3 or a colocalizing enzyme, 12R-LOX, to disruption of the normal permeability barrier of the skin [Jobard, F., Lefèvre, C., Karaduman, A., Blanchet-Bardon, C., Emre, S., Weissenbach, J., Özgüc, M., Lathrop, M., Prud'homme, J. F. & Fischer, J. (2002) Hum. Mol. Genet. 11, 107–113]. Herein we identify a logical link of the biochemistry to the genetics. eLOX3 functions as a hydroperoxide isomerase (epoxyalcohol synthase) by using the product of 12R-LOX as the preferred substrate. 12R-Hydroperoxyeicosatetraenoic acid (12R-HPETE) is converted to 8R-hydroxy-11R,12R-epoxyeicosa-5Z,9E,14Z-trienoic acid, one of the isomers of hepoxilin A3, and to 12-ketoeicosatetraenoic acid in a 2:1 ratio. Other hydroperoxides, including 8R-HPETE, 12S-HPETE, and 15S-HPETE, as well as the 13S- and 13R-hydroperoxides of linoleic acid are converted less efficiently. Mass spectrometric analysis of the epoxyalcohol formed from [18O]15S-HPETE showed that both hydroperoxy oxygens are retained in the product. We propose that the ferrous form of eLOX3 initiates a redox cycle, unprecedented among LOX in being autocatalytic, in which the hydroperoxy substrate is isomerized to the epoxyalcohol or keto product. Our results provide strong biochemical evidence for a functional linkage of 12R-LOX and eLOX3 and clues into skin biochemistry and the etiology of ichthyosiform diseases in humans. PMID:12881489

Lysyl Oxidase as a Serum Biomarker of Liver Fibrosis in Patients with Severe Obesity and Obstructive Sleep Apnea.

PubMed

Mesarwi, Omar A; Shin, Mi-Kyung; Drager, Luciano F; Bevans-Fonti, Shannon; Jun, Jonathan C; Putcha, Nirupama; Torbenson, Michael S; Pedrosa, Rodrigo P; Lorenzi-Filho, Geraldo; Steele, Kimberley E; Schweitzer, Michael A; Magnuson, Thomas H; Lidor, Anne O; Schwartz, Alan R; Polotsky, Vsevolod Y

2015-10-01

Obstructive sleep apnea (OSA) is associated with the progression of nonalcoholic fatty liver disease (NAFLD). We hypothesized that the hypoxia of OSA increases hepatic production of lysyl oxidase (LOX), an enzyme that cross-links collagen, and that LOX may serve as a biomarker of hepatic fibrosis. Thirty-five patients with severe obesity underwent liver biopsy, polysomnography, and serum LOX testing. A separate group with severe OSA had serum LOX measured before and after 3 mo of CPAP or no therapy, as did age-matched controls. LOX expression and secretion were measured in mouse hepatocytes following exposure to hypoxia. The Johns Hopkins Bayview Sleep Disorders Center, and the Hypertension Unit of the Heart Institute at the University of São Paulo Medical School. In the bariatric cohort, the apnea-hypopnea index was higher in patients with hepatic fibrosis than in those without fibrosis (42.7 ± 30.2 events/h, versus 16.2 ± 15.5 events/h; P = 0.002), as was serum LOX (84.64 ± 29.71 ng/mL, versus 45.46 ± 17.16 ng/mL; P < 0.001). In the sleep clinic sample, patients with severe OSA had higher baseline LOX than healthy controls (70.75 ng/mL versus 52.36 ng/mL, P = 0.046), and serum LOX decreased in patients with OSA on CPAP (mean decrease 20.49 ng/mL) but not in untreated patients (mean decrease 0.19 ng/mL). Hypoxic mouse hepatocytes demonstrated 5.9-fold increased LOX transcription (P = 0.046), and enhanced LOX protein secretion. The hypoxic stress of obstructive sleep apnea may increase circulating lysyl oxidase (LOX) levels. LOX may serve as a biomarker of liver fibrosis in patients with severe obesity and nonalcoholic fatty liver disease. © 2015 Associated Professional Sleep Societies, LLC.

The 9-lipoxygenase Osr9-LOX1 interacts with the 13-lipoxygenase-mediated pathway to regulate resistance to chewing and piercing-sucking herbivores in rice.

PubMed

Zhou, Guoxin; Ren, Nan; Qi, Jingfeng; Lu, Jing; Xiang, Caiyu; Ju, Hongping; Cheng, Jiaan; Lou, Yonggen

2014-09-01

Oxylipins produced by the 13-lipoxygenase (LOX) have been reported to play an important role in plant defense responses to herbivores. Yet, the role of oxylipins produced by the 9-LOX pathway in this process remains largely unknown. Here we cloned a gene encoding a chloroplast-localized 9-LOX, Osr9-LOX1, from rice. Transcriptional analysis revealed that herbivore infestation, mechanical wounding and jasmonic acid (JA) treatment either repressed or did not enhance the level of Osr9-LOX1 transcripts at early stages but did at later stages, whereas salicylic acid (SA) treatment quickly increased the transcript level of Osr9-LOX1. Antisense expression of Osr9-lox1 (as-r9lox1) decreased the amount of wound-induced (Z)-3-hexenal but increased levels of striped stem borer (SSB)-induced linolenic acid, JA, SA and trypsin protease inhibitors. These changes were associated with increased resistance in rice to the larvae of the SSB Chilo suppressalis. In contrast, although no significant differences were observed in the duration of the nymph stage or the number of eggs laid by female adults between the brown planthopper (BPH) Nilaparvata lugens that fed on as-r9lox1 lines and BPH that fed on wild-type (WT) rice plants, the survival rate of BPH nymphs that fed on as-r9lox1 lines was higher than that of nymphs that fed on WT plants, possibly because of a higher JA level. The results demonstrate that Osr9-LOX1 plays an important role in regulating an herbivore-induced JA burst and cross-talk between JA and SA, and in controlling resistance in rice to chewing and phloem-feeding herbivores. © 2014 Scandinavian Plant Physiology Society.

Root-expressed maize lipoxygenase 3 negatively regulates induced systemic resistance to Colletotrichum graminicola in shoots

PubMed Central

Constantino, Nasie N.; Mastouri, Fatemeh; Damarwinasis, Ramadhika; Borrego, Eli J.; Moran-Diez, Maria E.; Kenerley, Charley M.; Gao, Xiquan; Kolomiets, Michael V.

2013-01-01

We have previously reported that disruption of a maize root-expressed 9-lipoxygenase (9-LOX) gene, ZmLOX3, results in dramatic increase in resistance to diverse leaf and stalk pathogens. Despite evident economic significance of these findings, the mechanism behind this increased resistance remained elusive. In this study, we found that increased resistance of the lox3-4 mutants is due to constitutive activation of induced systemic resistance (ISR) signaling. We showed that ZmLOX3 lacked expression in leaves in response to anthracnose leaf blight pathogen Colletotrichum graminicola, but was expressed constitutively in the roots, thus, prompting our hypothesis: the roots of lox3-4 mutants are the source of increased resistance in leaves. Supporting this hypothesis, treatment of wild-type plants (WT) with xylem sap of lox3-4 mutant induced resistance to C. graminicola to the levels comparable to those observed in lox3-4 mutant. Moreover, treating mutants with the sap collected from WT plants partially restored the susceptibility to C. graminicola. lox3-4 mutants showed primed defense responses upon infection, which included earlier and greater induction of defense-related PAL and GST genes compared to WT. In addition to the greater expression of the octadecanoid pathway genes, lox3-4 mutant responded earlier and with a greater accumulation of H2O2 in response to C. graminicola infection or treatment with alamethicin. These findings suggest that lox3-4 mutants display constitutive ISR-like signaling. In support of this idea, root colonization by Trichoderma virens strain GV29-8 induced the same level of disease resistance in WT as the treatment with the mutant sap, but had no additional resistance effect in lox3-4 mutant. While treatment with T. virens GV29 strongly and rapidly suppressed ZmLOX3 expression in hydroponically grown WT roots, T. virens Δsml mutant, which is deficient in ISR induction, was unable to suppress expression of ZmLOX3, thus, providing genetic evidence that SM1 function in ISR, at least in part, by suppressing host ZmLOX3 gene. This study and the genetic tools generated herein will allow the identification of the signals regulating the induction of resistance to aboveground attackers by beneficial soil microorganisms in the future. PMID:24391653

Liquid Oxygen Rotating Friction Ignition Testing of Aluminum and Titanium with Monel and Inconel for Rocket Engine Propulsion System Contamination Investigation

NASA Technical Reports Server (NTRS)

Peralta, S.; Rosales, Keisa R.; Stoltzfus, Joel M.

2009-01-01

Metallic contaminant was found in the liquid oxygen (LOX) pre-valve screen of the shuttle main engine propulsion system on two orbiter vehicles. To investigate the potential for an ignition, NASA Johnson Space Center White Sands Test Facility performed (modified) rotating friction ignition testing in LOX. This testing simulated a contaminant particle in the low-pressure oxygen turbo pump (LPOTP) and the high-pressure oxygen turbo pump (HPOTP) of the shuttle main propulsion system. Monel(R) K-500 and Inconel(R) 718 samples represented the LPOTP and HPOTP materials. Aluminum foil tape and titanium foil represented the contaminant particles. In both the Monel(R) and Inconel(R) material configurations, the aluminum foil tape samples did not ignite after 30 s of rubbing. In contrast, all of the titanium foil samples ignited regardless of the rubbing duration or material configuration. However, the titanium foil ignitions did not propagate to the Monel and Inconel materials.

Tripropellant engine study

NASA Technical Reports Server (NTRS)

Wheeler, D. B.

1978-01-01

Engine performance data, combustion gas thermodynamic properties, and turbine gas parameters were determined for various high power cycle engine configurations derived from the space shuttle main engine that will allow sequential burning of LOX/hydrocarbon and LOX/hydrogen fuels. Both stage combustion and gas generator pump power cycles were considered. Engine concepts were formulated for LOX/RP-1, LOX/CH4, and LOX/C3H8 propellants. Flowrates and operating conditions were established for this initial set of engine systems, and the adaptability of the major components of shuttle main engine was investigated.

Tripropellant Engine Study

NASA Technical Reports Server (NTRS)

Wheeler, D. B.

1977-01-01

The feasibility of modifying the space shuttle main engine (SSME) for dual mode operation was investigated. Various high power cycle engine configurations derived from the SSME were configurations that will allow sequential burning of LOX/hydrocarbon and LOX/hydrogen were studied in order to identify concepts that make maximum use of SSME hardware and best satisfy the dual mode booster engine system application. Engine cycles were formulated for LOX/RP-1, LOX/CH4, and LOX/C3H8 propellants. Flow rates and operating cycles were established and the adaptability of the major components of the SSME was evaluated.

The lectin-like oxidized low-density lipoprotein receptor-1 as therapeutic target for atherosclerosis, inflammatory conditions and longevity.

PubMed

Ulrich-Merzenich, Gudrun; Zeitler, Heike

2013-08-01

The lectin-like oxidized LDL receptor-1 (LOX-1) is a scavenger receptor and is regarded as a central element in the initiation of endothelial dysfunction and its further progression to atherosclerosis. Increasing numbers of studies suggest that therapeutic strategies to modulate LOX-1 will have a broad spectrum of applications ranging from cardiovascular diseases to longevity. The dual role of LOX-1 as a culprit molecule in the process of atherosclerosis and as a danger signal in various tissues is introduced. The structure of the receptor, its ligands and its modulation by known drugs, by natural products (e.g., statins, imipramine, salicylate-based drugs, procyanidins, curcumin) and by new strategies (antisenseRNA, miRNA, pyrrole-imidazol-polyamides, LOX-1 antibodies, lipid apheresis) are described. Therapeutic approaches via transcript regulation, allowing a modulation of LOX-1, may be an easier and safer strategy than a blockade of the receptor. Considering the wide distribution of LOX-1 on different tissues, research on the mechanisms of LOX-1 modulation by drugs and natural products applying "omic"-technologies will not only allow a better understanding of the role of LOX-1 in the processes of atherosclerosis, inflammation and longevity but also support the development of specific LOX-1 modulators, avoiding the initiation of molecular mechanisms which lead to adverse events.

Evaluation of human LOX-12 as a serum marker for breast cancer.

PubMed

Singh, Abhay Kumar; Kant, Sashi; Parshad, Rajinder; Banerjee, Nirupama; Dey, Sharmistha

2011-10-22

The high concentration of prostaglandins has been associated with chronic inflammatory diseases and several types of human cancers. This is due to the over expression of inflammatory enzymes like Cyclooxygenase (COX), Lipoxygenase (LOX) etc. The aim of this study was to quantify the LOX-12 with clinicopathological parameter of breast cancer patients and its response after chemotherapy to establish serum LOX-12 as a prognostic marker. This case-controlled study was performed on 86 biopsy proven breast cancer patients. Blood and tissue samples were collected from the patients. Serum LOX-12 of the study group was quantified by Surface Plasmon Resonance (SPR) and ELISA techniques by antibody-antigen interaction strategy. A significant increase in LOX-12 levels was observed in breast cancer patients (Mean ± SD=40.54±13.61 ng/ml) as compared to healthy controls (Mean ± SD=13.42±2.4 ng/ml) (p<0.0001). Serum LOX-12 levels were significantly higher (p<0.002) in patients with lymph node involvement. More than 75% patients had shown significant (p<0.0001) reduction of LOX-12 levels after chemotherapy. This was also confirmed by ELISA. This study for the first time had co-related the quantity of serum LOX-12 with breast cancer and also with the effect of chemotherapy. Copyright © 2011 Elsevier Inc. All rights reserved.

Differential Induction of Lipoxygenase Isoforms in Wheat upon Treatment with Rust Fungus Elicitor, Chitin Oligosaccharides, Chitosan, and Methyl Jasmonate.

PubMed Central

Bohland, C.; Balkenhohl, T.; Loers, G.; Feussner, I.; Grambow, H. J.

1997-01-01

A glycopeptide elicitor prepared from germ tubes of the rust fungus Puccinia graminis Pers. f. sp. tritici Erikss. & Henn (Pgt), as well as chitin oligosaccharides, chitosan, and methyl jasmonate (MJ) stimulated lipoxygenase (LOX) activity (E.C. 1.13.11.12) in wheat (Triticum aestivum) leaves. Immunoblot analysis using anti-LOX antibodies revealed the induction of 92- and 103-kD LOX species after Pgt elicitor treatment. In contrast, MJ treatment led to a significant increase of a 100-kD LOX species, which was also detected at lower levels in control plants. The effects of chitin oligomers and chitosan resembled those caused by MJ. In conjunction with other observations the results suggest that separate reaction cascades exist, and that jasmonates may not be involved in Pgt elicitor action. LOX-92 appears to be mainly responsible for the increase in LOX activity after Pgt elicitor treatment because its appearance on western blots coincided with high LOX activity in distinct anion-exchange chromatography fractions. It is most active at pH 5.5 to 6.0, and product formation from linoleic and [alpha]-linolenic acid is clearly in favor of the 9-LOOHs. It is interesting that a 92-kD LOX species, which seems to correspond to the Pgt elicitor-induced LOX species, was also detected in rust-inoculated leaves. PMID:12223735

The use of programmable logic controllers (PLC) for rocket engine component testing

NASA Technical Reports Server (NTRS)

Nail, William; Scheuermann, Patrick; Witcher, Kern

1991-01-01

Application of PLCs to the rocket engine component testing at a new Stennis Space Center Component Test Facility is suggested as an alternative to dedicated specialized computers. The PLC systems are characterized by rugged design, intuitive software, fault tolerance, flexibility, multiple end device options, networking capability, and built-in diagnostics. A distributed PLC-based system is projected to be used for testing LH2/LOx turbopumps required for the ALS/NLS rocket engines.

In Vivo Imaging of mdrla Gene Expression

DTIC Science & Technology

2005-06-01

svImJ mouse strain, compatible with the ES cells used in our Transgenic Mouse Facility. b. Engineer PGK-neo and Renilla luciferase cassettes...inserted between the two loxP sites, upstream of the Neo cassette. A cloning strategy was then devised to fuse Renilla luciferase in-frame with the...sites: B, BamHI; E, EcoRI; S, ScaI. PGK-neo: neo under the control of the PGK promoter. Luc: Renilla luciferase fused in- frame with the translated

Environmental Assessment: Recapitalization of the 49th WG Combat Capabilities and Capacities, Holloman Air Force Base, New Mexico

DTIC Science & Technology

2011-07-01

of Liquid Oxygen (LOX). The new facilities would provide for the specific needs to maintain the F-16 engine, electronics , and maintenance... electronic countermeasures, and electronic counter countermeasures. 500 AGL to 40,000 MSL 0.5 to 1.0 hour Night Operations Aircraft intercepts (1...defensive maneuvering, chaff/flare, and electronic countermeasures. 2,000 AGL to 40,000 MSL 0.75 to 1.5 hour (Dissimilar) Air Combat Tactics

Conceptual Study of the LB/TS (Large Blast/Thermal Simulator) Instrumentation, Data Acquisition and Facility Controls System.

DTIC Science & Technology

1984-09-12

423, R. Jones ATTN: DAEN-ECE-T ATTN: DAEN-RDL DEPARTMENT OF THE AIR FORCE ATTN: DAEN-RDM. J. Healy ATTN: DAEN-ZCM Air Force Engineering & Services Ctr...31 7 Comparison Chart of Various Femperature Sensors (Prepared by HY-CAL Engineering ) ... ...... 36 8 Temperature Sensor Requirements and...Positions * TRS Valve Positions; LOX, AL, N2 Solenoid and Control . - Valve Positions 1.4 Air Compressor System Control * Valve Positions 0 Pressure

A chloroplast lipoxygenase is required for wound-induced jasmonic acid accumulation in Arabidopsis.

PubMed

Bell, E; Creelman, R A; Mullet, J E

1995-09-12

Plant lipoxygenases are thought to be involved in the biosynthesis of lipid-derived signaling molecules. The potential involvement of a specific Arabidopsis thaliana lipoxygenase isozyme, LOX2, in the biosynthesis of the plant growth regulators jasmonic acid (JA) and abscisic acid was investigated. Our characterization of LOX2 indicates that the protein is targeted to chloroplasts. The physiological role of this chloroplast lipoxygenase was analyzed in transgenic plants where cosuppression reduced LOX2 accumulation. The reduction in LOX2 levels caused no obvious changes in plant growth or in the accumulation of abscisic acid. However, the wound-induced accumulation of JA observed in control plants was absent in leaves of transgenic plants that lacked LOX2. Thus, LOX2 is required for the wound-induced synthesis of the plant growth regulator JA in leaves. We also examined the expression of a wound- and JA-inducible Arabidopsis gene, vsp, in transgenic and control plants. Leaves of transgenic plants lacking LOX2 accumulated less vsp mRNA than did control leaves in response to wounding. This result suggests that wound-induced JA (or some other LOX2-requiring component of the wound response pathway) is involved in the wound-induced regulation of this gene.

ALS LOX/H2 subscale coaxial injector testing

NASA Technical Reports Server (NTRS)

Dexter, Carol E.

1991-01-01

Tests of a 40K subscale LOX/H2 coaxial LOX swirl injector conducted without injector or chamber degradation are reported. Chamber pressures ranged from 1572 to 2355 psia with overall mixture ratios from 5.04 to 6.39. The highest characteristic velocities were measured when the mixture ratio across the injector face was uniform. Scarfing of the outer row LOX posts had the largest effect on chamber heating rates. As a result of the tests, the LSI design was modified to arrange the outer row LOX posts in a circular pattern, eliminate O/F biasing and fuel film cooling, and modify the interpropellant plate to allow for larger pressure differentials during the start and cutoff transients. Testing of a 100 K LOX/H2 coaxial LOX swirl injector involved chamber pressure ranging from 700 to 2500 psia with overall mixture ratios from 3.2 to 8.8. Stable combustion was observed to a fuel temperature of 90R and characteristic velocity efficiencies were good.

Space shuttle orbit maneuvering engine reusable thrust chamber

NASA Technical Reports Server (NTRS)

1972-01-01

A data dump is presented containing space shuttle orbiter maneuvering engine performance, weight, envelope, and interface pressure requirements for candidate propellant combinations (NTO/MMH, NTO50-50, LOX/MMH, LOX/50-50, LOX/N2H4, LOX/C3H8, and LOX/RP-1) and cooling concepts (regenerative and dump/film). These data are presented parametrically for the thrust, chamber pressure, nozzle expansion ratio, and engine mixture ratio ranges of interest. Also included is information describing sensitivity to system changes; reliability, maintainability and safety; development programs and associated critical technology areas; engine cost comparisons during development and operation; and ecological effects.

Clinical analysis of lectin-like oxidized low-density lipoprotein receptor-1 in patients with in-stent restenosis after percutaneous coronary intervention.

PubMed

Liu, Junfeng; Liu, Yunde; Jia, Kegang; Huo, Zhixiao; Huo, Qianyu; Liu, Zhili; Li, Yongshu; Han, Xuejing; Wang, Rong

2018-04-01

In-stent restenosis (ISR) is the most common complication associated with percutaneous coronary intervention (PCI). Although some studies have reported an association between lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) and ISR, not enough clinical validation data are available to support this link. Here, we report our cross-sectional study aimed at exploring the feasibility of LOX-1 as a biomarker for the prognostic diagnosis of patients undergoing PCI.Three groups were included: ISR group, including 99 patients with ISR diagnosed with coronary arteriography (CAG) after PCI; lesion group, comprising 87 patients with coronary artery stenosis (<50%) diagnosed with CAG after PCI; and control group, consisting of 96 volunteers with no coronary artery disease. The levels of LOX-1 were measured in each patient by using an enzyme-linked immunosorbent assay, and their general information as well as laboratory parameters were recorded and followed up during a period of 2 years.LOX-1 levels gradually increased after PCI along with the progression of the lesion in the 3 groups. The levels of LOX-1 were significantly higher in the ISR group than in the other 2 groups (P < .001). LOX-1 levels were correlated with the levels of uric acid (UA) (r = 0.289, P = .007), creatinine (CREA) (r = .316, P = .003), and high-density lipoprotein cholesterol (HDL-C) (r = -0.271, P = .012), whereas no statistically significant correlation was detected with the Gensini score (r = 0.157, P = .141). The sensitivity and specificity of LOX-1 were 81.5% and 55.7%, respectively, with the most optimal threshold (5.04 μg/L). The area under curve (AUC) of the receiver operator characteristic (ROC) curve of LOX-1 was 0.720, and LOX-1 had the highest AUC compared with CREA, UA, and HDL-C, both individually and in combination.A high level of LOX-1 in the early period after PCI has a certain predictive power and diagnostic value for ISR. However, the level of LOX-1 is not related to the Gensini score of coronary artery after PCI, and CREA and UA, which are weakly related to LOX-1, have no obvious synergy in the diagnosis of ISR with LOX-1.

Lectin-like oxidized LDL receptor-1 expresses in mouse bone marrow-derived mesenchymal stem cells and stimulates their proliferation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Fenxi; Stem Cell and Biotheraphy Technology Research Center, College of Lifescience and Technology, Xinxiang Medical University, Xinxiang 453003; Wang, Congrui

The bone marrow-derived mesenchymal stem cells (bmMSCs) have been widely used in cell transplant therapy, and the proliferative ability of bmMSCs is one of the determinants of the therapy efficiency. Lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) as a transmembrane protein is responsible for binding, internalizing and degrading oxidized low density lipoprotein (ox-LDL). It has been identified that LOX-1 is expressed in endothelial cells, vascular smooth muscle cells, cardiomyocytes, fibroblasts and monocytes. In these cells, low concentration of ox-LDL (<40 μg/mL) stimulates their proliferation via LOX-1 activation. However, it is poor understood that whether LOX-1 is expressed in bmMSCs andmore » which role it plays. In this study, we investigated the status of LOX-1 expression in bmMSCs and its function on bmMSC proliferation. Our results showed that primary bmMSCs exhibiting a typical fibroblast-like morphology are positive for CD44 and CD90, but negative for CD34 and CD45. LOX-1 in both mRNA and protein levels is highly expressed in bmMSCs. Meanwhile, bmMSCs exhibit a strong potential to take up ox-LDL. Moreover, LOX-1 expression in bmMSCs is upregulated by ox-LDL with a dose- and time-dependent manner. Presence of ox-LDL also enhances the proliferation of bmMSCs. Knockdown of LOX-1 expression significantly inhibits ox-LDL-induced bmMSC proliferation. These findings indicate that LOX-1 plays a role in bmMSC proliferation. - Highlights: ► LOX-1 expresses in bmMSCs and mediates uptake of ox-LDL. ► Ox-LDL stimulates upregulation of LOX-1 in bmMSCs. ► Ox-LDL promotes bmMSC proliferation and expression of Mdm2, phosphor-Akt, phosphor-ERK1/2 and phosphor-NF-κB. ► LOX-1 siRNA inhibits ox-LDL-induced bmMSC proliferation and expression cell survival signals.« less

Impact of myeloperoxidase-derived oxidants on the product profile of human 5-lipoxygenase.

PubMed

Zschaler, Josefin; Dorow, Juliane; Schöpe, Louisa; Ceglarek, Uta; Arnhold, Jürgen

2015-08-01

Human 5-lipoxygenase (5-LOX) oxidizes arachidonic acid to 5S-hydroperoxy-6 E,8 Z,11 Z,14 Z-eicosatetraenoic acid (5-HpETE) and leukotriene (LT) A4. In neutrophils, LTA4 is further converted to the potent chemoattractant LTB4. These cells also contain the heme enzyme myeloperoxidase (MPO), which produces several potent oxidants such as hypochlorous acid (HOCl), which are involved in pathogen defense and immune regulation. Here, we addressed the question whether MPO-derived oxidants are able to affect the activity of 5-LOX and the product profile of this enzyme. Human 5-LOX was incubated with increasing amounts of HOCl or HOBr. Afterward, arachidonic acid metabolites of 5-LOX were analyzed by reverse-phase high-performance liquid chromatography as well as by liquid chromatography-electrospray ionization-tandem mass spectrometry. The incubation of 5-LOX with the MPO-derived oxidants significantly changed the product profile of 5-LOX. Thereby, HOCl and HOBr increased the ratio of 5-H(p)ETE to 6-trans-LTB4 in a concentration-dependent manner. At low oxidant concentrations, there was a strong decrease in the yield of 6-trans-LTB4, whereas 5-HpETE did not change or increased. Additionally, the formation of 8-HpETE and 12-HpETE by 5-LOX rose slightly with increasing HOCl and HOBr. Comparable results were obtained with the MPO-H2O2-Cl(-) system when glucose oxidase and glucose were applied as a source of H2O2. This was necessary because of a strong impairment of 5-LOX activity by H2O2. In summary, MPO-derived oxidants showed a considerable impact on 5-LOX, impairing the epoxidation of 5-HpETE, whereas the hydroperoxidation of arachidonic acid was unaffected. Apparently, this was caused by an oxidative modification of critical amino acid residues of 5-LOX. Further work is necessary to assess the specific type and position of oxidation in the substrate-binding cavity of 5-LOX and to specify whether this interaction between 5-LOX and MPO-derived oxidants also takes place in stimulated neutrophils. Copyright © 2015 Elsevier Inc. All rights reserved.

Swirl Coaxial Injector Testing with LOX/RP-J

NASA Technical Reports Server (NTRS)

Greene, Sandra Elam; Casiano, Matt

2013-01-01

Testing was conducted at NASA fs Marshall Space Flight Center (MSFC) in the fall of 2012 to evaluate the operation and performance of liquid oxygen (LOX) and kerosene (RP ]1) in an existing swirl coaxial injector. While selected Russian engines use variations of swirl coaxial injectors, component level performance data has not been readily available, and all previously documented component testing at MSFC with LOX/RP ]1 had been performed using a variety of impinging injector designs. Impinging injectors have been adequate for specific LOX/RP ]1 engine applications, yet swirl coaxial injectors offer easier fabrication efforts, providing cost and schedule savings for hardware development. Swirl coaxial elements also offer more flexibility for design changes. Furthermore, testing with LOX and liquid methane propellants at MSFC showed that a swirl coaxial injector offered improved performance compared to an impinging injector. So, technical interest was generated to see if similar performance gains could be achieved with LOX/RP ]1 using a swirl coaxial injector. Results would allow such injectors to be considered for future engine concepts that require LOX/RP ]1 propellants. Existing injector and chamber hardware was used in the test assemblies. The injector had been tested in previous programs at MSFC using LOX/methane and LOX/hydrogen propellants. Minor modifications were made to the injector to accommodate the required LOX/RP ]1 flows. Mainstage tests were performed over a range of chamber pressures and mixture ratios. Additional testing included detonated gbombs h for stability data. Test results suggested characteristic velocity, C*, efficiencies for the injector were 95 ]97%. The injector also appeared dynamically stable with quick recovery from the pressure perturbations generated in the bomb tests.

On the Role of Molecular Oxygen in Lipoxygenase Activation

PubMed Central

Zheng, Yuxiang; Brash, Alan R.

2010-01-01

The oxygenation of polyunsaturated fatty acids by lipoxygenases (LOX) is associated with a lag phase during which the resting ferrous enzyme is converted to the active ferric form by reaction with fatty acid hydroperoxide. Epidermal lipoxygenase-3 (eLOX3) is atypical in displaying hydroperoxide isomerase activity with fatty acid hydroperoxides through cycling of the ferrous enzyme. Yet eLOX3 is capable of dioxygenase activity, albeit with a long lag phase and need for high concentrations of hydroperoxide activator. Here, we show that higher O2 concentration shortens the lag phase in eLOX3, although it reduces the rate of hydroperoxide consumption, effects also associated with an A451G mutation known to affect the disposition of molecular oxygen in the LOX active site. These observations are consistent with a role of O2 in interrupting hydroperoxide isomerase cycling. Activation of eLOX3, A451G eLOX3, and soybean LOX-1 with 13-hydroperoxy-linoleic acid forms oxygenated end products, which we identified as 9R- and 9S-hydroperoxy-12S,13S-trans-epoxyoctadec-10E-enoic acids. We deduce that activation partly depends on reaction of O2 with the intermediate of hydroperoxide cleavage, the epoxyallylic radical, giving an epoxyallylic peroxyl radical that does not further react with Fe(III)-OH; instead, it dissociates and leaves the enzyme in the activated free ferric state. eLOX3 differs from soybean LOX-1 in more tightly binding the epoxyallylic radical and having limited access to O2 within the active site, leading to a deficiency in activation and a dominant hydroperoxide isomerase activity. PMID:20923767

Association between lysyl oxidase and fibrotic focus in relation with inflammation in breast cancer

PubMed Central

Jeong, Young Ju; Park, Sung Hwan; Mun, Sung Hee; Kwak, Sang Gyu; Lee, Sun-Jae; Oh, Hoon Kyu

2018-01-01

We hypothesized that lysyl oxidase (LOX) contributes to the formation of fibrotic focus (FF) in association with inflammation and serves a significant role in breast carcinogenesis. In the present study, the association between the expression of LOX family members and FF with regards to with inflammation was analyzed, and the prognostic significance of LOX and FF in breast cancer was investigated. Immunohistochemical staining for LOX, LOX-like protein (LOXL) 1, LOXL2 and LOXL3 was performed in primary breast cancer tissues. The status of FF within the tumor was assessed, including size and grade. Levels of inflammatory markers, intratumoral and peritumoral lymphocyte infiltration were also evaluated. The clinicopathological characteristics were evaluated from the medical records of patients. In the present study, the expression of LOX family members was not associated with the presence of FF. FF was identified to be associated with intratumoral and peritumoral inflammation, tumor stage, larger tumor size, lymph node metastasis, high histologic grade, and p53 expression. LOX and LOXL3 were associated with intratumoral, and peritumoral inflammation. Furthermore, LOXL1 was associated with intratumoral inflammation and interleukin-4. In addition, LOX was associated with cluster of differentiation 8+ T cells. LOXL3 was associated with expression of ER and PR, and molecular subtype. In the survival analysis, overall survival time was statistically significantly longer in the FF-negative compared with that in the FF-positive group. In conclusion, it was demonstrated that FF and the expression of LOX family members were associated with inflammation in breast cancer. FF was associated with poor prognostic markers of breast cancer. Further studies are required to clarify the mechanisms underlying the association between the LOX family, FF and inflammation in breast cancer. PMID:29434955

Cyanobacterial Lactate Oxidases Serve as Essential Partners in N2 Fixation and Evolved into Photorespiratory Glycolate Oxidases in Plants[w

PubMed Central

Hackenberg, Claudia; Kern, Ramona; Hüge, Jan; Stal, Lucas J.; Tsuji, Yoshinori; Kopka, Joachim; Shiraiwa, Yoshihiro; Bauwe, Hermann; Hagemann, Martin

2011-01-01

Glycolate oxidase (GOX) is an essential enzyme involved in photorespiratory metabolism in plants. In cyanobacteria and green algae, the corresponding reaction is catalyzed by glycolate dehydrogenases (GlcD). The genomes of N2-fixing cyanobacteria, such as Nostoc PCC 7120 and green algae, appear to harbor genes for both GlcD and GOX proteins. The GOX-like proteins from Nostoc (No-LOX) and from Chlamydomonas reinhardtii showed high l-lactate oxidase (LOX) and low GOX activities, whereas glycolate was the preferred substrate of the phylogenetically related At-GOX2 from Arabidopsis thaliana. Changing the active site of No-LOX to that of At-GOX2 by site-specific mutagenesis reversed the LOX/GOX activity ratio of No-LOX. Despite its low GOX activity, No-LOX overexpression decreased the accumulation of toxic glycolate in a cyanobacterial photorespiratory mutant and restored its ability to grow in air. A LOX-deficient Nostoc mutant grew normally in nitrate-containing medium but died under N2-fixing conditions. Cultivation under low oxygen rescued this lethal phenotype, indicating that N2 fixation was more sensitive to O2 in the Δlox Nostoc mutant than in the wild type. We propose that LOX primarily serves as an O2-scavenging enzyme to protect nitrogenase in extant N2-fixing cyanobacteria, whereas in plants it has evolved into GOX, responsible for glycolate oxidation during photorespiration. PMID:21828292

Molecular cloning of a ripening-specific lipoxygenase and its expression during wild-type and mutant tomato fruit development.

PubMed Central

Kausch, K D; Handa, A K

1997-01-01

A 94-kD protein that accumulates predominately in tomato (Ly-copersicon esculentum) fruit during ripening was purified, and antibodies specific for the purified protein were used to isolate cDNA clones from a red-ripe fruit cDNA library. A sequence analysis of these cDNAs and cross-reactivity of the 94-kD-specific antibodies to the soybean lipoxygenase (LOX) L-1, L-2, and L-3 proteins and soybean LOX L-1-specific antibodies to the 94-kD protein identified it as a member of the LOX gene family. Maximum levels of the 94-kD LOX mRNA and protein are present in breaker to ripe and red-ripe stages, respectively. Expression of 94-kD LOX in different tissues from mature green and red-ripe tomato fruits was found to be greatest in the radial walls of ripe fruit, but immunocytolocalization using tissue printing suggests that the highest accumulation of its protein occurs in locular jelly. None of 94-kD LOX is expressed in nonripening mutant fruits of any age. Never-ripe mutant fruit accumulate the 94-kD LOX mRNA to levels similar to those obtained in wild-type fruit, but fail to accumulate the 94-kD LOX protein. Collectively, the results show that expression of 94-kD LOX is regulated by the ripening process, and ethylene may play a role in its protein accumulation. PMID:9112767

Lectin-Like Oxidized LDL Receptor-1 Is an Enhancer of Tumor Angiogenesis in Human Prostate Cancer Cells

PubMed Central

González-Chavarría, Iván; Cerro, Rita P.; Parra, Natalie P.; Sandoval, Felipe A.; Zuñiga, Felipe A.; Omazábal, Valeska A.; Lamperti, Liliana I.; Jiménez, Silvana P.; Fernandez, Edelmira A.; Gutiérrez, Nicolas A.; Rodriguez, Federico S.; Onate, Sergio A.; Sánchez, Oliberto; Vera, Juan C.; Toledo, Jorge R.

2014-01-01

Altered expression and function of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) has been associated with several diseases such as endothelial dysfunction, atherosclerosis and obesity. In these pathologies, oxLDL/LOX-1 activates signaling pathways that promote cell proliferation, cell motility and angiogenesis. Recent studies have indicated that olr1 mRNA is over-expressed in stage III and IV of human prostatic adenocarcinomas. However, the function of LOX-1 in prostate cancer angiogenesis remains to be determined. Our aim was to analyze the contribution of oxLDL and LOX-1 to tumor angiogenesis using C4-2 prostate cancer cells. We analyzed the expression of pro-angiogenic molecules and angiogenesis on prostate cancer tumor xenografts, using prostate cancer cell models with overexpression or knockdown of LOX-1 receptor. Our results demonstrate that the activation of LOX-1 using oxLDL increases cell proliferation, and the expression of the pro-angiogenic molecules VEGF, MMP-2, and MMP-9 in a dose-dependent manner. Noticeably, these effects were prevented in the C4-2 prostate cancer model when LOX-1 expression was knocked down. The angiogenic effect of LOX-1 activated with oxLDL was further demonstrated using the aortic ring assay and the xenograft model of tumor growth on chorioallantoic membrane of chicken embryos. Consequently, we propose that LOX-1 activation by oxLDL is an important event that enhances tumor angiogenesis in human prostate cancer cells. PMID:25170920

Composite load spectra for select space propulsion structural components

NASA Technical Reports Server (NTRS)

Newell, J. F.; Kurth, R. E.; Ho, H.

1986-01-01

A multiyear program is performed with the objective to develop generic load models with multiple levels of progressive sophistication to simulate the composite (combined) load spectra that are induced in space propulsion system components, representative of Space Shuttle Main Engines (SSME), such as transfer ducts, turbine blades, and liquid oxygen (LOX) posts. Progress of the first year's effort includes completion of a sufficient portion of each task -- probabilistic models, code development, validation, and an initial operational code. This code has from its inception an expert system philosophy that could be added to throughout the program and in the future. The initial operational code is only applicable to turbine blade type loadings. The probabilistic model included in the operational code has fitting routines for loads that utilize a modified Discrete Probabilistic Distribution termed RASCAL, a barrier crossing method and a Monte Carlo method. An initial load model was developed by Battelle that is currently used for the slowly varying duty cycle type loading. The intent is to use the model and related codes essentially in the current form for all loads that are based on measured or calculated data that have followed a slowly varying profile.

A chloroplast lipoxygenase is required for wound-induced jasmonic acid accumulation in Arabidopsis.

PubMed Central

Bell, E; Creelman, R A; Mullet, J E

1995-01-01

Plant lipoxygenases are thought to be involved in the biosynthesis of lipid-derived signaling molecules. The potential involvement of a specific Arabidopsis thaliana lipoxygenase isozyme, LOX2, in the biosynthesis of the plant growth regulators jasmonic acid (JA) and abscisic acid was investigated. Our characterization of LOX2 indicates that the protein is targeted to chloroplasts. The physiological role of this chloroplast lipoxygenase was analyzed in transgenic plants where cosuppression reduced LOX2 accumulation. The reduction in LOX2 levels caused no obvious changes in plant growth or in the accumulation of abscisic acid. However, the wound-induced accumulation of JA observed in control plants was absent in leaves of transgenic plants that lacked LOX2. Thus, LOX2 is required for the wound-induced synthesis of the plant growth regulator JA in leaves. We also examined the expression of a wound- and JA-inducible Arabidopsis gene, vsp, in transgenic and control plants. Leaves of transgenic plants lacking LOX2 accumulated less vsp mRNA than did control leaves in response to wounding. This result suggests that wound-induced JA (or some other LOX2-requiring component of the wound response pathway) is involved in the wound-induced regulation of this gene. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 PMID:7567995

Cannabidiol-2',6'-dimethyl ether as an effective protector of 15-lipoxygenase-mediated low-density lipoprotein oxidation in vitro.

PubMed

Takeda, Shuso; Hirayama, Akari; Urata, Shino; Mano, Nobutaka; Fukagawa, Keiko; Imamura, Midori; Irii, Ayumi; Kitajima, Satomi; Masuyama, Tomoko; Nomiyama, Mai; Tatei, Sachiko; Tomita, Saari; Kudo, Taichi; Noguchi, Momoko; Yamaguchi, Yasuhiro; Okamoto, Yoshiko; Amamoto, Toshiaki; Fukunishi, Yoshifumi; Watanabe, Kazuhito; Omiecinski, Curtis John; Aramaki, Hironori

2011-01-01

15-Lipoxygenase (15-LOX) is one of the key enzymes responsible for the formation of oxidized low-density lipoprotein (ox-LDL), a major causal factor for atherosclerosis. Both enzymatic (15-LOX) and non-enzymatic (Cu(2+)) mechanisms have been proposed for the production of ox-LDL. We have recently reported that cannabidiol-2',6'-dimethyl ether (CBDD) is a selective and potent inhibitor of 15-LOX-catalyzed linoleic acid oxygenation (Takeda et al., Drug Metab. Dispos., 37, 1733-1737 (2009)). In the LDL, linoleic acid is present as cholesteryl linoleate, the major fatty acid esterified to cholesterol, and is susceptible to oxidative modification by 15-LOX or Cu(2+). In this investigation, we examined the efficacy of CBDD on i) 15-LOX-catalyzed oxygenation of cholesteryl linoleate, and ii) ox-LDL formation catalyzed by 15-LOX versus Cu(2+)-mediated non-enzymatic generation of this important mediator. The results obtained demonstrate that CBDD is a potent and selective inhibitor of ox-LDL formation generated by the 15-LOX pathway. These studies establish CBDD as both an important experimental tool for characterizing 15-LOX-mediated ox-LDL formation, and as a potentially useful therapeutic agent for treatment of atherosclerosis.

Cannabidiol-2′,6′-dimethyl Ether as an Effective Protector of 15-Lipoxygenase-Mediated Low-Density Lipoprotein Oxidation in Vitro

PubMed Central

Takeda, Shuso; Hirayama, Akari; Urata, Shino; Mano, Nobutaka; Fukagawa, Keiko; Imamura, Midori; Irii, Ayumi; Kitajima, Satomi; Masuyama, Tomoko; Nomiyama, Mai; Tatei, Sachiko; Tomita, Saari; Kudo, Taichi; Noguchi, Momoko; Yamaguchi, Yasuhiro; Okamoto, Yoshiko; Amamoto, Toshiaki; Fukunishi, Yoshifumi; Watanabe, Kazuhito; Omiecinski, Curtis John; Aramaki, Hironori

2014-01-01

15-Lipoxygenase (15-LOX) is one of the key enzymes responsible for the formation of oxidized low-density lipoprotein (ox-LDL), a major causal factor for atherosclerosis. Both enzymatic (15-LOX) and non-enzymatic (Cu2+) mechanisms have been proposed for the production of ox-LDL. We have recently reported that cannabidiol-2′,6′-dimethyl ether (CBDD) is a selective and potent inhibitor of 15-LOX-catalyzed linoleic acid oxygenation (Takeda et al., Drug Metab. Dispos., 37, 1733–1737 (2009)). In the LDL, linoleic acid is present as cholesteryl linoleate, the major fatty acid esterified to cholesterol, and is susceptible to oxidative modification by 15-LOX or Cu2+. In this investigation, we examined the efficacy of CBDD on i) 15-LOX-catalyzed oxygenation of cholesteryl linoleate, and ii) ox-LDL formation catalyzed by 15-LOX versus Cu2+-mediated non-enzymatic generation of this important mediator. The results obtained demonstrate that CBDD is a potent and selective inhibitor of ox-LDL formation generated by the 15-LOX pathway. These studies establish CBDD as both an important experimental tool for characterizing 15-LOX-mediated ox-LDL formation, and as a potentially useful therapeutic agent for treatment of atherosclerosis. PMID:21804214

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gilbert, Nathaniel C.; Rui, Zhe; Neau, David B.

The enzyme 5-lipoxygenase (5-LOX) initiates biosynthesis of the proinflammatory leukotriene lipid mediators and, together with 15-LOX, is also required for synthesis of the anti-inflammatory lipoxins. The catalytic activity of 5-LOX is regulated through multiple mechanisms, including Ca{sup 2+}-targeted membrane binding and phosphorylation at specific serine residues. To investigate the consequences of phosphorylation at S663, we mutated the residue to the phosphorylation mimic Asp, providing a homogenous preparation suitable for catalytic and structural studies. The S663D enzyme exhibits robust 15-LOX activity, as determined by spectrophotometric and HPLC analyses, with only traces of 5-LOX activity remaining; synthesis of the anti-inflammatory lipoxin A4more » from arachidonic acid is also detected. The crystal structure of the S663D mutant in the absence and presence of arachidonic acid (in the context of the previously reported Stable-5-LOX) reveals substantial remodeling of helices that define the active site so that the once fully encapsulated catalytic machinery is solvent accessible. Our results suggest that phosphorylation of 5-LOX at S663 could not only down-regulate leukotriene synthesis but also stimulate lipoxin production in inflammatory cells that do not express 15-LOX, thus redirecting lipid mediator biosynthesis to the production of proresolving mediators of inflammation.« less

One vs two primary LOX feedline configuration study for the National Launch System

NASA Technical Reports Server (NTRS)

Dill, K.; Davis, D.; Bates, R.; Tarwater, R.

1992-01-01

Six single LOX feedline designs were evaluated for use on the National Launch Vehicle. A single feedline design, designated the 'Spider', was chosen and compared to the baseline system. The baseline configuration employs two 20-inch I.D. lines, each supplying LOX to three 650,000 lbf thrust Space Transportation Main Engines. Five single feedline diameters were examined for the spider configuration; 22, 24, 26, 28, and 30-inch I.D. System dry weights and LOX residuals were estimated. These parameters, along with calculated staged mass for the different single line and baseline configurations, were used to calculate the payload mass to orbit. For the cases where LOX is drained to minimum NPSP conditions, none of the single lines performed as well as the dual line system, although the 22-inch diameter single line compared well. However, for the cases where LOX is drained to operating levels (LOX level at the booster and spider manifolds for the dual and single line configurations, respectively), the 22 - 26-inch I.D. single line systems show a greater payload capability.

LC/ESR/MS study of pH-dependent radical generation from 15-LOX catalyzed DPA peroxidation

PubMed Central

Purwaha, Preeti; Gu, Yan; Kelavkar, Uddhav; Kang, Jing Xuan; Law, Benedict; Wu, Erxi; Qian, Steven Y.

2011-01-01

Docosapentaenoic acid (DPA) is a unique fatty acid that exists in two isomeric forms (n-3 and n-6) which differ in their physiological behaviors. DPA can undergo free-radical mediated peroxidation via lipoxygenase (LOX). 15-LOX, one of the LOX isomers, has received much attention in cancer research due to its very different expression level in normal tissues compared to tumors and some bioactive fatty acid metabolites modulating the tumorigenic pathways in cancer. However, the mechanism linking 15-LOX, DPA-metabolites, and the bioactivities is still unclear, and the free radicals generated in DPA peroxidation have never been characterized. In this study, we have studied radicals formed from both soybean and human cellular (PC3-15LOS cells) 15-LOX-catalyzed peroxidation of DPAs at different pH’s using a combination of LC/ESR/MS with the spin trapping technique. We observed a total of three carbon-centered radicals formed in 15-LOX/DPA (n-3) stemming from its 7-, 17- and 20-hydroperoxides, while only one formed from 17-hydroperoxide in DPA (n-6). A change in the reaction pH from 8.5 (15-LOX enzyme optimum) to 7.4 (physiological) and to 6.5 (tumor, acidic) not only decreased the total radical formation but also altered the preferred site of oxygenation. This pH-dependent alteration of radical formation and oxygenation pattern may have significant implications and provide a basis for our ongoing investigations of LOXs as well as fatty acids in cancer biology. PMID:21807091

Air Launch: Examining Performance Potential of Various Configurations and Growth Options

NASA Technical Reports Server (NTRS)

Waters, Eric D.; Creech, Dennis M.; Philips, Alan

2013-01-01

The Advanced Concepts Office at NASA's George C. Marshall Space Flight Center conducted a high-level analysis of various air launch vehicle configurations, objectively determining maximum launch vehicle payload while considering carrier aircraft capabilities and given dimensional constraints. With the renewed interest in aerial launch of low-earth orbit payloads, referenced by programs such as Stratolaunch and Spaceship2, there existed a need to qualify the boundaries of the trade space, identify performance envelopes, and understand advantages and limiting factors of designing for maximum payload capability. Using the NASA/DARPA Horizontal Launch Study (HLS) Point Design 2 (PD-2) as a point-of-departure configuration, two independent design actions were undertaken. Both configurations utilized a Boeing 747-400F as the carrier aircraft, LOX/RP-1 first stage and LOX/LH2 second stage. Each design was sized to meet dimensional and mass constraints while optimizing propellant loads and stage delta V (?V) splits. All concepts, when fully loaded, exceeded the allowable Gross Takeoff Weight (GTOW) of the aircraft platform. This excess mass was evaluated as propellant/fuel offload available for a potential in-flight refueling scenario. Results indicate many advantages such as large, relative payload delivery of approximately 47,000 lbm and significant mission flexibility, such as variable launch site inclination and launch window; however, in-flight cryogenic fluid transfer and carrier aircraft platform integration are substantial technical hurdles to the realization of such a system configuration.

Engine system assessment study using Martian propellants

NASA Technical Reports Server (NTRS)

Pelaccio, Dennis; Jacobs, Mark; Scheil, Christine; Collins, John

1992-01-01

A top-level feasibility study was conducted that identified and characterized promising chemical propulsion system designs which use two or more of the following propellant combinations: LOX/H2, LOX/CH4, and LOX/CO. The engine systems examined emphasized the usage of common subsystem/component hardware where possible. In support of this study, numerous mission scenarios were characterized that used various combinations of Earth, lunar, and Mars propellants to establish engine system requirements to assess the promising engine system design concept examined, and to determine overall exploration leverage of such systems compared to state-of-the-art cryogenic (LOX/H2) propulsion systems. Initially in the study, critical propulsion system technologies were assessed. Candidate expander and gas generator cycle LOX/H2/CO, LOX/H2/CH4, and LOX/CO/CH4 engine system designs were parametrically evaluated. From this evaluation baseline, tripropellant Mars Transfer Vehicle (MTV) LOX cooled and bipropellant Lunar Excursion Vehicle (LEV) and Mars Excursion Vehicle (MEV) engine systems were identified. Representative tankage designs for a MTV were also investigated. Re-evaluation of the missions using the baseline engine design showed that in general the slightly lower performance, smaller, lower weight gas generator cycle-based engines required less overall mission Mars and in situ propellant production (ISPP) infrastructure support compared to the larger, heavier, higher performing expander cycle engine systems.

Retention and Molecular Evolution of Lipoxygenase Genes in Modern Rosid Plants

PubMed Central

Chen, Zhu; Chen, Danmei; Chu, Wenyuan; Zhu, Dongyue; Yan, Hanwei; Xiang, Yan

2016-01-01

Whole-genome duplication events have occurred more than once in the genomes of some rosids and played a significant role over evolutionary time. Lipoxygenases (LOXs) are involved in many developmental and resistance processes in plants. Our study concerns the subject of the LOX gene family; we tracked the evolutionary process of ancestral LOX genes in four modern rosids. Here we show that some members of the LOX gene family in the Arabidopsis genome are likely to be lost during evolution, leading to a smaller size than that in Populus, Vitis, and Carica. Strong purifying selection acted as a critical role in almost all of the paralogous and orthologous genes. The structure of LOX genes in Carica and Populus are relatively stable, whereas Vitis and Arabidopsis have a difference. By searching conserved motifs of LOX genes, we found that each sub-family shared similar components. Research on intraspecies gene collinearity show that recent duplication holds an important position in Populus and Arabidopsis. Gene collinearity analysis within and between these four rosid plants revealed that all LOX genes in each modern rosid were the offspring from different ancestral genes. This study traces the evolution of LOX genes which have been differentially retained and expanded in rosid plants. Our results presented here may aid in the selection of special genes retained in the rosid plants for further analysis of biological function. PMID:27746812

LOX-1 activation by oxLDL triggers an epithelial mesenchymal transition and promotes tumorigenic potential in prostate cancer cells.

PubMed

González-Chavarría, I; Fernandez, E; Gutierrez, N; González-Horta, E E; Sandoval, F; Cifuentes, P; Castillo, C; Cerro, R; Sanchez, O; Toledo, Jorge R

2018-02-01

Obesity is related to an increased risk of developing prostate cancer with high malignancy stages or metastasis. Recent results demonstrated that LOX-1, a receptor associated with obesity and atherosclerosis, is overexpressed in advanced and metastatic prostate cancer. Furthermore, high levels of oxLDL, the main ligand for LOX-1, have been found in patients with advanced prostate cancer. However, the role of LOX-1 in prostate cancer has not been unraveled completely yet. Here, we show that LOX-1 is overexpressed in prostate cancer cells and its activation by oxLDL promotes an epithelial to mesenchymal transition, through of lowered expression of epithelial markers (E-cadherin and plakoglobin) and an increased expression of mesenchymal markers (vimentin, N-cadherin, snail, slug, MMP-2 and MMP-9). Consequently, LOX-1 activation by oxLDL promotes actin cytoskeleton restructuration and MMP-2 and MMP-9 activity inducing prostate cancer cell invasion and migration. Additionally, LOX-1 increased the tumorigenic potential of prostate cancer cells and its expression was necessary for tumor growth in nude mice. In conclusion, our results suggest that oxLDL/LOX-1 could be ones of mechanisms that explain why obese patients with prostate cancer have an accelerated tumor progression and a greater probability of developing metastasis. Copyright © 2017 Elsevier B.V. All rights reserved.

Crystal structure of a lipoxygenase from Cyanothece sp. may reveal novel features for substrate acquisition[S

PubMed Central

Newie, Julia; Andreou, Alexandra; Neumann, Piotr; Einsle, Oliver; Feussner, Ivo; Ficner, Ralf

2016-01-01

In eukaryotes, oxidized PUFAs, so-called oxylipins, are vital signaling molecules. The first step in their biosynthesis may be catalyzed by a lipoxygenase (LOX), which forms hydroperoxides by introducing dioxygen into PUFAs. Here we characterized CspLOX1, a phylogenetically distant LOX family member from Cyanothece sp. PCC 8801 and determined its crystal structure. In addition to the classical two domains found in plant, animal, and coral LOXs, we identified an N-terminal helical extension, reminiscent of the long α-helical insertion in Pseudomonas aeruginosa LOX. In liposome flotation studies, this helical extension, rather than the β-barrel domain, was crucial for a membrane binding function. Additionally, CspLOX1 could oxygenate 1,2-diarachidonyl-sn-glycero-3-phosphocholine, suggesting that the enzyme may act directly on membranes and that fatty acids bind to the active site in a tail-first orientation. This binding mode is further supported by the fact that CspLOX1 catalyzed oxygenation at the n-10 position of both linoleic and arachidonic acid, resulting in 9R- and 11R-hydroperoxides, respectively. Together these results reveal unifying structural features of LOXs and their function. While the core of the active site is important for lipoxygenation and thus highly conserved, peripheral domains functioning in membrane and substrate binding are more variable. PMID:26667668

15-Lipoxygenase-1 Production is Lost in Pancreatic Cancer and Overexpression of the Gene Inhibits Tumor Cell Growth1

PubMed Central

Hennig, René; Kehl, Timo; Noor, Seema; Ding, Xian-Zhong; Rao, Sambasiva M; Bergmann, Frank; Fürstenberger, Gerhard; Büchler, Markus W; Friess, Helmut; Krieg, Peter; Adrian, Thomas E

2007-01-01

Pancreatic cancer patients have an abysmal prognosis because of late diagnosis and lack of therapeutic options. Pancreatic intraepithelial neoplasias (PanINs), the precursor lesions, are a potential target for chemoprevention. Targeting eicosanoid pathways is an obvious choice because 5-lipoxygenase (5-LOX) has been suggested as a tumor promoter in pancreatic carcinogenesis. Here we provide evidence that 15-lipoxygenase-1 (15-LOX-1) expression and activity may exert antitumorigenic effects in pancreatic cancer. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis showed absence or very weak expression of 15-LOX-1 in all pancreatic cancer cell lines tested. 15-LOX-1 was strongly stained in normal ductal cells, tubular complexes, and centroacinar cells, but no staining was seen in islets, cancer cells, PanIN lesions, or in tumor cells in lymph node metastases, indicating that 15-LOX-1 expression is lost during tumor development in human pancreas. Overexpression of 15-LOX-1 in pancreatic tumor cells or treatment with its arachidonic acid-derived metabolite resulted in decreased cell growth. These findings provide evidence that loss of 15-LOX-1 may play an important role in pancreatic carcinogenesis, possibly as a tumor suppressor gene. Thus, induction of 15-LOX-1 expression may be an attractive option for the prevention and treatment of pancreatic cancer. PMID:18030360

Lysyl oxidase propeptide inhibits smooth muscle cell signaling and proliferation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hurtado, Paola A.; Vora, Siddharth; Sume, Siddika Selva

2008-02-01

Lysyl oxidase is required for the normal biosynthesis and maturation of collagen and elastin. It is expressed by vascular smooth muscle cells, and its increased expression has been previously found in atherosclerosis and in models of balloon angioplasty. The lysyl oxidase propeptide (LOX-PP) has more recently been found to have biological activity as a tumor suppressor, and it inhibits Erk1/2 Map kinase activation. We reasoned that LOX-PP may have functions in normal non-transformed cells. We, therefore, investigated its effects on smooth muscle cells, focusing on important biological processes mediated by Erk1/2-dependent signaling pathways including proliferation and matrix metalloproteinase-9 (MMP-9) expression.more » In addition, we investigated whether evidence for accumulation of LOX-PP could be found in vivo in a femoral artery injury model. Recombinant LOX-PP was expressed and purified, and was found to inhibit primary rat aorta smooth muscle cell proliferation and DNA synthesis by more than 50%. TNF-{alpha}-stimulated MMP-9 expression and Erk1/2 activation were both significantly inhibited by LOX-PP. Immunohistochemistry studies carried out with affinity purified anti-LOX-PP antibody showed that LOX-PP epitopes were expressed at elevated levels in vascular lesions of injured arteries. These novel data suggest that LOX-PP may provide a feedback control mechanism that serves to inhibit properties associated with the development of vascular pathology.« less

A Global Assessment of Circulating Prolysyl Oxidase in Nonischemic Patients With Garden-variety Heart Failure With Preserved Ejection Fraction.

PubMed

Muñoz Calvo, Benjamín; Villa Martínez, Ana; López Orgil, Susana; López Andrés, Natalia; Román García, Feliciano; Víctor Palomares, Virginia; de la Calle de la Villa, Esther; Nadador Patiño, Verónica; Arribas-Gómez, Ignacio

2018-05-25

Lysyl oxidase is overexpressed in the myocardium of patients with hypertensive cardiomyopathy. We aimed to explore whether patients with hypertensive-metabolic heart failure with preserved ejection fraction (HM-HFpEF) also have increased concentrations of circulating prolysyl oxidase (cpLOX) and its possible consequences. We quantified cpLOX concentrations in 85 nonischemic patients with stage C, HM-HFpEF, and compared them with those of 51 healthy controls. We also assessed the correlations of cpLOX with myocardial stiffness parameters, collagen turnover products and fibrogenic cytokines, as well as the predictive value of plasma proenzyme levels at 1-year of follow-up. We detected raised cpLOX values and found that they correlated with calculated E/E' ratios and stiffness constants. The subgroup of patients with type I diastolic dysfunction showed a single negative correlation between cpLOX and B-type natriuretic peptide whereas patients with a restrictive diastolic pattern showed a strong correlation between cpLOX and galectin-3. Kaplan-Meier analysis revealed that cpLOX > 52.20 ng/mL slightly increased the risk of a fatal outcome (log-rank = 4.45; P = .034). When Cox regression was used, cpLOX was found to be a significant independent predictor of cardiovascular death or hospitalization due to the decompensation of HM-HFpEF (HR, 1.360; 95%CI, 1.126-1.638; P = .046). Patients with symptomatic HM-HFpEF show high cpLOX serum levels associated with restrictive diastolic filling indices. These levels represent a moderate risk factor for poor clinical outcome. Throughout the natural history of HM-HFpEF, we observed that cpLOX concentrations were initially negatively correlated with B-type natriuretic peptide but positively correlated with galectin-3 as advanced diastolic dysfunction developed. Copyright © 2018. Published by Elsevier España, S.L.U.

T cell protein tyrosine phosphatase (TCPTP) deficiency in muscle does not alter insulin signalling and glucose homeostasis in mice.

PubMed

Loh, K; Merry, T L; Galic, S; Wu, B J; Watt, M J; Zhang, S; Zhang, Z-Y; Neel, B G; Tiganis, T

2012-02-01

Insulin activates insulin receptor protein tyrosine kinase and downstream phosphatidylinositol-3-kinase (PI3K)/Akt signalling in muscle to promote glucose uptake. The insulin receptor can serve as a substrate for the protein tyrosine phosphatase (PTP) 1B and T cell protein tyrosine phosphatase (TCPTP), which share a striking 74% sequence identity in their catalytic domains. PTP1B is a validated therapeutic target for the alleviation of insulin resistance in type 2 diabetes. PTP1B dephosphorylates the insulin receptor in liver and muscle to regulate glucose homeostasis, whereas TCPTP regulates insulin receptor signalling and gluconeogenesis in the liver. In this study we assessed for the first time the role of TCPTP in the regulation of insulin receptor signalling in muscle. We generated muscle-specific TCPTP-deficient (Mck-Cre;Ptpn2(lox/lox)) mice (Mck, also known as Ckm) and assessed the impact on glucose homeostasis and muscle insulin receptor signalling in chow-fed versus high-fat-fed mice. Blood glucose and insulin levels, insulin and glucose tolerance, and insulin-induced muscle insulin receptor activation and downstream PI3K/Akt signalling remained unaltered in chow-fed Mck-Cre;Ptpn2(lox/lox) versus Ptpn2(lox/lox) mice. In addition, body weight, adiposity, energy expenditure, insulin sensitivity and glucose homeostasis were not altered in high-fat-fed Mck-Cre;Ptpn2(lox/lox) versus Ptpn2(lox/lox) mice. These results indicate that TCPTP deficiency in muscle has no effect on insulin signalling and glucose homeostasis, and does not prevent high-fat diet-induced insulin resistance. Thus, despite their high degree of sequence identity, PTP1B and TCPTP contribute differentially to insulin receptor regulation in muscle. Our results are consistent with the notion that these two highly related PTPs make distinct contributions to insulin receptor regulation in different tissues.

Down-regulation of lipoxygenase gene reduces degradation of carotenoids of golden rice during storage.

PubMed

Gayen, Dipak; Ali, Nusrat; Sarkar, Sailendra Nath; Datta, Swapan K; Datta, Karabi

2015-07-01

Down-regulation of lipoxygenase enzyme activity reduces degradation of carotenoids of bio-fortified rice seeds which would be an effective tool to reduce huge post-harvest and economic losses of bio-fortified rice seeds during storage. Bio-fortified provitamin A-enriched rice line (golden rice) expressing higher amounts of β-carotene in the rice endosperm provides vitamin A for human health. However, it is already reported that degradation of carotenoids during storage is a major problem. The gene responsible for degradation of carotenoids during storage has remained largely unexplored till now. In our previous study, it has been shown that r9-LOX1 gene is responsible for rice seed quality deterioration. In the present study, we attempted to investigate if r9-LOX1 gene has any role in degradation of carotenoids in rice seeds during storage. To establish our hypothesis, the endogenous lipoxygenase (LOX) activity of high-carotenoid golden indica rice seed was silenced by RNAi technology using aleurone layer and embryo-specific Oleosin-18 promoter. To check the storage stability, LOX enzyme down-regulated high-carotenoid T3 transgenic rice seeds were subjected to artificial aging treatment. The results obtained from biochemical assays (MDA, ROS) also indicated that after artificial aging, the deterioration of LOX-RNAi lines was considerably lower compared to β-carotene-enriched transgenic rice which had higher LOX activity in comparison to LOX-RNAi lines. Furthermore, it was also observed by HPLC analysis that down-regulation of LOX gene activity decreases co-oxidation of β-carotene in LOX-RNAi golden rice seeds as compared to the β-carotene-enriched transgenic rice, after artificial aging treatment. Therefore, our study substantially establishes and verifies that LOX is a key enzyme for catalyzing co-oxidation of β-carotene and has a significant role in deterioration of β-carotene levels in the carotenoid-enriched golden rice.

Overexpression and Down-Regulation of Barley Lipoxygenase LOX2.2 Affects Jasmonate-Regulated Genes and Aphid Fecundity

PubMed Central

Losvik, Aleksandra; Beste, Lisa; Glinwood, Robert; Ivarson, Emelie; Stephens, Jennifer; Zhu, Li-Hua; Jonsson, Lisbeth

2017-01-01

Aphids are pests on many crops and depend on plant phloem sap as their food source. In an attempt to find factors improving plant resistance against aphids, we studied the effects of overexpression and down-regulation of the lipoxygenase gene LOX2.2 in barley (Hordeum vulgare L.) on the performance of two aphid species. A specialist, bird cherry-oat aphid (Rhopalosiphum padi L.) and a generalist, green peach aphid (Myzus persicae Sulzer) were studied. LOX2.2 overexpressing lines showed up-regulation of some other jasmonic acid (JA)-regulated genes, and antisense lines showed down-regulation of such genes. Overexpression or suppression of LOX2.2 did not affect aphid settling or the life span on the plants, but in short term fecundity tests, overexpressing plants supported lower aphid numbers and antisense plants higher aphid numbers. The amounts and composition of released volatile organic compounds did not differ between control and LOX2.2 overexpressing lines. Up-regulation of genes was similar for both aphid species. The results suggest that LOX2.2 plays a role in the activation of JA-mediated responses and indicates the involvement of LOX2.2 in basic defense responses. PMID:29257097

Challenging Pneumatic Requirements for Acoustic Testing of the Cryogenic Second Stage for the New Delta 3 Rocket

NASA Technical Reports Server (NTRS)

Webb, Andrew T.

1998-01-01

The paper describes the unique pneumatic test requirements for the acoustic and shock separation testing of the Second Stage for the new Delta III Rocket at the Goddard Space Flight Center in Greenbelt, Maryland. The testing was conducted in the 45,000 cu ft (25-feet wide by 30-feet deep by 50-foot high) Acoustic Facility. The acoustic testing required that the liquid oxygen (LOX) and liquid hydrogen (LH2) tanks be filled with enough liquid nitrogen (LN2) to simulate launch fuel masses during testing. The challenge for this test dealt with designing, procuring, and fabricating the pneumatic supply systems for quick assembly while maintaining the purity requirements and minimizing costs. The pneumatic systems were designed to fill and drain the both LOX and LH2 tanks as well as to operate the fill/drain and vent valves for each of the tanks. The test criteria for the pneumatic sub-systems consisted of function, cleanliness, availability, and cost. The first criteria, function, required the tanks to be filled and drained in an efficient manner while preventing them from seeing pressures greater than 9 psig which would add a pressure cycle to the tank. An LN2 tanker, borrowed from another NASA facility, served as the pre-cool and drain tanker. Pre-cooling the tanks allowed for more efficient and cost effective transfer from the LN2 delivery tankers. Helium gas, supplied from a high purity tube trailer, was used to pressurize the vapor space above the LN2 pushing it into the drain tanker. The tube trailer also supplied high pressure helium to the vehicle for valve control and component purges. Cleanliness was maintained by proper component selection, end-use particle filtration, and any on-site cleaning determined necessary by testing. In order to meet the availability/cost juggling act, products designed for LOX delivery systems were procured to ensure system compatibility while off the shelf valves and tubing designed for the semiconductor industry were procured for the gas systems.

Structural and functional basis of phospholipid oxygenase activity of bacterial lipoxygenase from Pseudomonas aeruginosa.

PubMed

Banthiya, Swathi; Kalms, Jacqueline; Galemou Yoga, Etienne; Ivanov, Igor; Carpena, Xavi; Hamberg, Mats; Kuhn, Hartmut; Scheerer, Patrick

2016-11-01

Pseudomonas aeruginosa expresses a secreted LOX-isoform (PA-LOX, LoxA) capable of oxidizing polyenoic fatty acids to hydroperoxy derivatives. Here we report high-level expression of this enzyme in E. coli and its structural and functional characterization. Recombinant PA-LOX oxygenates polyenoic fatty acids including eicosapentaenoic acid and docosahexaenoic acid to the corresponding (n-6)S-hydroperoxy derivatives. This reaction involves abstraction of the proS-hydrogen from the n-8 bisallylic methylene. PA-LOX lacks major leukotriene synthase activity but converts 5S-HETE and 5S,6R/S-DiHETE to anti-inflammatory and pro-resolving lipoxins. It also exhibits phospholipid oxygenase activity as indicated by the formation of a specific pattern of oxygenation products from different phospholipid subspecies. Multiple mutagenesis studies revealed that PA-LOX does not follow classical concepts explaining the reaction specificity of mammalian LOXs. The crystal structure of PA-LOX was solved with resolutions of up to 1.48Å and its polypeptide chain is folded as single domain. The substrate-binding pocket consists of two fatty acid binding subcavities and lobby. Subcavity-1 contains the catalytic non-heme iron. A phosphatidylethanolamine molecule occupies the substrate-binding pocket and its sn1 fatty acid is located close to the catalytic non-heme iron. His377, His382, His555, Asn559 and the C-terminal Ile685 function as direct iron ligands and a water molecule (hydroxyl) completes the octahedral ligand sphere. Although the biological relevance of PA-LOX is still unknown its functional characteristics (lipoxin synthase activity) implicate this enzyme in a bacterial evasion strategy aimed at downregulating the hosts' immune system. Copyright © 2016 Elsevier B.V. All rights reserved.

Adenovirus-mediated truncated Bid overexpression induced by the Cre/LoxP system promotes the cell apoptosis of CD133+ ovarian cancer stem cells.

PubMed

Long, Qifang; Yang, Ru; Lu, Weixian; Zhu, Weipei; Zhou, Jundong; Zheng, Cui; Zhou, Dongmei; Yu, Ling; Wu, Jinchang

2017-01-01

Cancer stem cells are a small subset of cancer cells that contribute to cancer progression, metastasis, chemoresistance and recurrence. CD133-positive (CD133+) ovarian cancer cells have been identified as ovarian cancer stem cells. Adenovirus-mediated gene therapy is an innovative therapeutic method for cancer treatment. In the present study, we aimed to develop a new gene therapy to specifically eliminate CD133+ ovarian cancer stem cells by targeting CD133. We used the Cre/LoxP system to augment the selective expression of the truncated Bid (tBid) gene as suicide gene therapy in CD133+ ovarian cancer stem cells. The adenovirus (Ad)-CD133-Cre expressing Cre recombinase under the control of the CD133 promoter and Ad-CMV-LoxP-Neo-LoxP-tBid expressing tBid under the control of the CMV promoter were successfully constructed using the Cre/LoxP switching system. The co-infection of Ad-CMV-LoxP-Neo-LoxP-tBid and Ad-CD133-Cre selectively induced tBid overexpression, which inhibited cell growth and triggered the cell apoptosis of CD133+ ovarian cancer stem cells. The Cre/LoxP system-mediated tBid overexpression activated the pro-apoptotic signaling pathway and augmented the cytotoxic effect of cisplatin in CD133+ ovarian cancer stem cells. Furthermore, in xenograft experiments, co-infection with the two recombinant adenoviruses markedly suppressed tumor growth in vivo and promoted cell apoptosis in tumor tissues. Taken together, the present study provides evidence that the adenovirus-mediated tBid overexpression induced by the Cre/LoxP system can effectively eliminate CD133+ ovarian cancer stem cells, representing a novel therapeutic strategy for the treatment of ovarian cancer.

GATA3 Inhibits Lysyl Oxidase Mediated Metastases of Human Basal Triple-Negative Breast Cancer Cells

PubMed Central

Chu, Isabel M.; Michalowski, Aleksandra M.; Hoenerhoff, Mark; Szauter, Kornelia M.; Luger, Dror; Sato, Misako; Flanders, Kathy; Oshima, Akira; Csiszar, Katalin; Green, Jeffrey E.

2011-01-01

Discovery of mechanisms that impede the aggressive and metastatic phenotype of human basal triple-negative type breast cancers (BTNBC) could provide novel targets for therapy for this form of breast cancer that has a relatively poor prognosis. Previous studies have demonstrated that the expression of GATA3, the master transcriptional regulator of mammary luminal differentiation, can reduce the tumorigenicity and metastatic propensity of the human BTNBC MDA-MB-231 cell line (MB231), although the mechanism for reduced metastases was not elucidated. We demonstrate through gene expression profiling that GATA3 expression in 231 cells resulted in the dramatic reduction in the expression of Lysyl oxidase (LOX), a metastasis-promoting matrix remodeling protein, in part, through methylation of the LOX promoter. Suppression of LOX expression by GATA3 was further confirmed in the BTNBC Hs578T cell line. Conversely, reduction of GATA3 expression by siRNA in luminal BT474 cells increased LOX expression. Reconstitution of LOX expression in 231-GATA3 cells restored metastatic propensity. A strong inverse association between high LOX and low GATA3 expression was confirmed in a panel of 51 human breast cancer cell lines. Similarly, human breast cancer microarray data demonstrated that high LOX/low GATA3 expression is associated with the BTNBC subtype of breast cancer and poor patient prognosis. Expression of GATA3 reprograms BTNBC to a less aggressive phenotype and inhibits a major mechanism of metastasis through inhibition of LOX. Induction of GATA3 in BTNBC cells or novel approaches that inhibit LOX expression or activity could be important strategies for treating BTNBC. PMID:21892208

Insulin-like growth factor I reduces lipid oxidation and foam cell formation via downregulation of 12/15-lipoxygenase.

PubMed

Sukhanov, Sergiy; Snarski, Patricia; Vaughn, Charlotte; Lobelle-Rich, Patricia; Kim, Catherine; Higashi, Yusuke; Shai, Shaw-Yung; Delafontaine, Patrice

2015-02-01

We have shown that insulin-like growth factor I (IGF-1) infusion in Apoe(-/-) mice decreased atherosclerotic plaque size and plaque macrophage and lipid content suggesting that IGF-1 suppressed formation of macrophage-derived foam cells. Since 12/15-lipoxygenase (12/15-LOX) plays an important role in OxLDL and foam cell formation, we hypothesized that IGF-1 downregulates 12/15-LOX, thereby suppressing lipid oxidation and foam cell formation. We found that IGF-1 decreased 12/15-LOX plaque immunopositivity and serum OxLDL levels in Apoe(-/-) mice. IGF-1 reduced 12/15-LOX protein and mRNA levels in cultured THP-1 macrophages and IGF-1 also decreased expression of STAT6 transcription factor. IGF-1 reduction in macrophage 12/15-LOX was mediated in part via a PI3 kinase- and STAT6-dependent transcriptional mechanism. IGF-1 suppressed THP-1 macrophage ability to oxidize lipids and form foam cells. IGF-1 downregulated 12/15-LOX in human blood-derived primary macrophages and IGF-1 decreased LDL oxidation induced by these cells. IGF-1 reduced LDL oxidation and formation of foam cells by wild type murine peritoneal macrophages, however these effects were completely blocked in 12/15-LOX-null macrophages suggesting that the ability of IGF-1 to reduce LDL oxidation and foam cells formation is dependent on its ability to downregulate 12/15-LOX. Overall our data demonstrate that IGF-1 reduces lipid oxidation and foam cell formation via downregulation of 12/15-LOX and this mechanism may play a major role in the anti-atherosclerotic effects of IGF-1. Published by Elsevier Ireland Ltd.

Insulin-like Growth Factor I Reduces Lipid Oxidation and Foam Cell Formation via Downregulation of 12/15-lipoxygenase

PubMed Central

Sukhanov, Sergiy; Snarski, Patricia; Vaughn, Charlotte; Lobelle-Rich, Patricia; Kim, Catherine; Higashi, Yusuke; Shai, Shaw-Yung; Delafontaine, Patrice

2014-01-01

Objective We have shown that insulin-like growth factor I (IGF-1) infusion in Apoe−/− mice decreased atherosclerotic plaque size and plaque macrophage and lipid content suggesting that IGF-1 suppressed formation of macrophage-derived foam cells. Since 12/15-lipoxygenase (12/15-LOX) plays an important role in OxLDL and foam cell formation, we hypothesized that IGF-1 downregulates 12/15-LOX, thereby suppressing lipid oxidation and foam cell formation. Approach and Results We found that IGF-1 decreased 12/15-LOX plaque immunopositivity and serum OxLDL levels in Apoe−/− mice. IGF-1 reduced 12/15-LOX protein and mRNA levels in cultured THP-1 macrophages and IGF-1 also decreased expression of STAT6 transcription factor. IGF-1 reduction in macrophage 12/15-LOX was mediated in part via a PI3 kinase- and STAT6-dependent transcriptional mechanism. IGF-1 suppressed THP-1 macrophage ability to oxidize lipids and form foam cells. IGF-1 downregulated 12/15-LOX in human blood-derived primary macrophages and IGF-1 decreased LDL oxidation induced by these cells. IGF-1 reduced LDL oxidation and formation of foam cells by wild type murine peritoneal macrophages, however these effects were completely blocked in 12/15-LOX-null macrophages suggesting that the ability of IGF-1 to reduce LDL oxidation and foam cells formation is dependent on its ability to downregulate 12/15-LOX. Conclusions Overall our data demonstrate that IGF-1 reduces lipid oxidation and foam cell formation via downregulation of 12/15-LOX and this mechanism may play a major role in the anti-atherosclerotic effects of IGF-1. PMID:25549319

Lysyl oxidase interacts with AGE signalling to modulate collagen synthesis in polycystic ovarian tissue

PubMed Central

Papachroni, Katerina K; Piperi, Christina; Levidou, Georgia; Korkolopoulou, Penelope; Pawelczyk, Leszek; Diamanti-Kandarakis, Evanthia; Papavassiliou, Athanasios G

2010-01-01

Abstract Connective tissue components – collagen types I, III and IV – surrounding the ovarian follicles undergo drastic changes during ovulation. Abnormal collagen synthesis and increased volume and density of ovarian stroma characterize the polycystic ovary syndrome (PCOS). During the ovulatory process, collagen synthesis is regulated by prolyl hydroxylase and lysyl oxidase (LOX) activity in ovarian follicles. LOX catalyzes collagen and elastin cross-linking and plays essential role in coordinating the control of ovarian extracellular matrix (ECM) during follicular development. We have recently shown accumulation of advanced glycation end products (AGEs), molecules that stimulate ECM production and abnormal collagen cross-linking, in ovarian tissue. However, the possible link between LOX and AGEs-induced signalling in collagen production and stroma formation in ovarian tissue from PCOS remains elusive. The present study investigates the hypothesis of AGE signalling pathway interaction with LOX gene activity in polycystic ovarian (PCO) tissue. We show an increased distribution and co-localization of LOX, collagen type IV and AGE molecules in the PCO tissue compared to control, as well as augmented expression of AGE signalling mediators/effectors, phospho(p)-ERK, phospho(p)-c-Jun and nuclear factor κB (NF-κB) in pathological tissue. Moreover, we demonstrate binding of AGE-induced transcription factors, NF-κB and activator protein-1 (AP-1) on LOX promoter, indicating a possible involvement of AGEs in LOX gene regulation, which may account for the documented increase in LOX mRNA and protein levels compared to control. These findings suggest that deposition of excess collagen in PCO tissue that induces cystogenesis may, in part, be due to AGE-mediated stimulation of LOX activity. PMID:19583806

Homocysteine facilitates LOX-1 activation and endothelial death through the PKCβ and SIRT1/HSF1 mechanism: relevance to human hyperhomocysteinaemia.

PubMed

Hung, Ching-Hsia; Chan, Shih-Hung; Chu, Pei-Ming; Tsai, Kun-Ling

2015-09-01

HHcy (hyperhomocysteinaemia) is one of the major risk factors for cardiovascular diseases. A high concentration of Hcy (homocysteine) induces endothelial dysfunction by activating endothelial oxidative stress. LOX-1 (lectin-like oxidized low-density lipoprotein receptor 1) plays a vital role in regulating the progression of atherosclerotic lesions. LOX-1 activation causes endothelial apoptosis and inflammation. The mechanism is still unclear as to whether Hcy affects human endothelial LOX-1 expression. LOX-1 expression level was confirmed by Western blotting assay in Hcy-treated endothelial cells. L-Methionine was used for HHcy induction in animals. Our results suggested that Hcy increased PKCβ (protein kinase Cβ) activation to enhance the LOX-1 expression level. The up-regulation of PKCβ phosphorylation subsequently causes ROS (reactive oxygen species) formation and SIRT1 (sirtuin 1) degradation through a proteasome-dependent mechanism, thereby mitigating the activity of SIRT1 by deacetylating HSF1 (heat-shock transcription factor 1). We also found that NOX2 is a key NAPDH oxidase isoform responsible for the Hcy-caused ROS formation. The overexpression of SIRT1 and HSF1 reduced the Hcy-induced LOX-1 activation. Silencing PKCβ function also reduced LOX-1 activation and endothelial apoptosis caused by Hcy. Our hypothesis was supported by analysing the data from methionine-induced HHcy-affected animals. Our data indicate a new direction for LOX-1 regulation by the modulation of the PKCβ/NAPDH oxidase/SIRT1/HSF1 mechanism. Our findings might provide a novel route for developing new therapeutic treatments for HHcy. © 2015 Authors; published by Portland Press Limited.

Pioglitazone inhibits LOX-1 expression in human coronary artery endothelial cells by reducing intracellular superoxide radical generation.

PubMed

Mehta, Jawahar L; Hu, Bo; Chen, Jiawei; Li, Dayuan

2003-12-01

LOX-1, a novel lectin-like receptor for oxidized LDL (ox-LDL), is expressed in response to ox-LDL, angiotensin II (Ang II), tumor necrosis factor (TNF)-alpha, and other stress stimuli. It is highly expressed in atherosclerotic tissues. Peroxisome proliferator-activated receptor (PPAR)-gamma ligands, such as pioglitazone, exert antiatherosclerotic effects. This study examined the regulation of LOX-1 expression in human coronary artery endothelial cells (HCAECs) by pioglitazone. Fourth generation HCAECs were treated with ox-LDL, Ang II, or TNF-alpha with or without pioglitazone pretreatment. All 3 stimuli upregulated LOX-1 expression (mRNA and protein). Pioglitazone, in a concentration-dependent manner, reduced LOX-1 expression (P<0.01 versus ox-LDL, Ang II, or TNF-alpha alone). Ox-LDL, Ang II, and TNF-alpha each enhanced intracellular superoxide radical generation, and pioglitazone pretreatment reduced superoxide generation (P<0.01 versus ox-LDL, Ang II, or TNF-alpha). Furthermore, all 3 stimuli upregulated the expression of the transcription factors nuclear factor-kappaB and activator protein-1 (determined by electrophoretic mobility shift assay), and pioglitazone pretreatment reduced this expression (P<0.01 versus ox-LDL, Ang II, or TNF-alpha). To determine the biological significance of pioglitazone-mediated downregulation of LOX-1, we studied monocyte adhesion to ox-LDL-treated HCAECs. Pioglitazone reduced the adhesion of monocytes to activated HCAECs in a fashion similar to that produced by antisense to LOX-1 mRNA. These observations suggest that the PPAR-gamma ligand pioglitazone reduces intracellular superoxide radical generation and subsequently reduces the expression of transcription factors, expression of the LOX-1 gene, and monocyte adhesion to activated endothelium. The salutary effect of PPAR-gamma ligands in atherogenesis may involve the inhibition of LOX-1 and the adhesion of monocytes to endothelium.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhao, Yu; Wang, Wenhui; Wang, Qi

Highlights: Black-Right-Pointing-Pointer 5-LOX is able to upregulate expression of NF-{kappa}B p65. Black-Right-Pointing-Pointer 5-LOX enhances nuclear translocation of NF-{kappa}B p65 via increasing p-I{kappa}B-{alpha} level. Black-Right-Pointing-Pointer 5-LOX stimulates transcriptional activity of NF-{kappa}B in hepatoma cells. Black-Right-Pointing-Pointer LTB4 activates transcriptional activity of NF-{kappa}B in hepatoma cells. -- Abstract: The issue that lipid metabolism enzyme and its metabolites regulate transcription factors in cancer cell is not fully understood. In this study, we first report that the lipid metabolism enzyme 5-Lipoxygenase (5-LOX) and its metabolite leukotriene B4 (LTB4) are capable of activating nuclear factor-{kappa}B (NF-{kappa}B) in hepatoma cells. We found that the treatment of MK886more » (an inhibitor of 5-LOX) or knockdown of 5-LOX was able to downregulate the expression of NF-{kappa}B p65 at the mRNA level and decreased the phosphorylation level of inhibitor {kappa}B{alpha} (I{kappa}B{alpha}) in the cytoplasm of hepatoma HepG2 or H7402 cells, which resulted in the decrease of the level of nuclear NF-{kappa}B p65. These were confirmed by immunofluorescence staining in HepG2 cell. Moreover, the above treatments were able to decrease the transcriptional activity of NF-{kappa}B in the cells. The LTB4, one of metabolites of 5-LOX, is responsible for 5-LOX-activated NF-{kappa}B in a dose-dependent manner. Thus, we conclude that the lipid metabolism enzyme 5-LOX and its metabolite LTB4 are capable of activating transcription factor NF-{kappa}B in hepatoma cells. Our finding provides new insight into the significance of lipid metabolism in activation of transcription factors in cancer.« less

Photographic Combustion Characterization of LOX/Hydrocarbon Type Propellants

NASA Technical Reports Server (NTRS)

Judd, D. C.

1980-01-01

The advantages and limitations of using high speed photography to identify potential combustion anomalies (pops, fuel freezing, reactive stream separation (RSS), carbon formation) were demonstrated. Combustion evaluation criteria were developed for evaluating, characterizing, and screening promising low cost propellant combination(s) and injector element(s) for long life, reusable engine systems. Carbon formation and RSS mechanisms and trends were identified by using high speed color photography at speeds up to 6000 frames/sec. Single element injectors were tested with LOX/RP-1, LOX/Propane, LOX/Methane and LOX/Ammonia propellants. Tests were conducted using seven separate injector elements. Five different conventionally machined elements were tested: OFO Triplet; Rectangular Unlike Doublet (RUD); Unlike Doublet (UD); Like on Lke Doublet (LOL-EDM); and Slit Triplet.

Dual nozzle design update. [on liquid rocket engines for advanced earth-to-orbit transportation systems

NASA Technical Reports Server (NTRS)

Obrien, C. J.

1982-01-01

Dual-nozzle engines, such as the dual-throat and dual-expander engines, are being evaluated for advanced earth-to-orbit transportation systems. Potential derivatives of the Space Shuttle and completely new vehicles might benefit from these advanced engines. In this paper, progress in the design of single-fuel and dual-fuel dual-nozzle engines is summarized. Dual-nozzle engines include those burning propellants such as LOX/RP-1/LH2, LOX/LC3H8/LH2, LOX/LCH4/LH2, LOX/LH2/LH2, LOX/LCH4/LCH4, LOX/LC3H8/C3H8 and N2O4/MMH/LH2. Engine data are applicable for thrust levels from 200,000 through 670,000 lbF. The results indicate that several versions of these engines utilize state-of-the-art technology and that even advanced versions of these engines do not require a major breakthrough in technology.

LOX Gene Transcript Accumulation in Olive (Olea europaea L.) Fruits at Different Stages of Maturation: Relationship between Volatile Compounds, Environmental Factors, and Technological Treatments for Oil Extraction

PubMed Central

Muzzalupo, Innocenzo; Macchione, Barbara; Bucci, Cristina; Stefanizzi, Francesca; Perri, Enzo; Chiappetta, Adriana; Tagarelli, Antonio; Sindona, Giovanni

2012-01-01

The quality of olive oil is influenced by genetic and environmental factors and by the maturation state of drupes, but it is equally affected by technological treatments of the process. This work investigates the possible correlation between olive LOX gene transcript accumulation, evaluated in fruits collected at different stages of maturation, and chemical biomarkers of its activity. During olive fruit ripening, the same genotype harvested from two different farms shows a positive linear trend between LOX relative transcript accumulation and the content of volatile compounds present in the olive oil aroma. Interestingly, a negative linear trend was observed between LOX relative transcript accumulation and the content of volatile compounds present in the olive pastes obtained from olive fruits with and without malaxation. The changes in the olive LOX transcript accumulation reveal its environmental regulation and suggest differential physiological functions for the LOXs. PMID:22645430

Evaporation of LOX under supercritical and subcritical conditions

NASA Technical Reports Server (NTRS)

Yang, A. S.; Hsieh, W. H.; Kuo, K. K.; Brown, J. J.

1993-01-01

The evaporation of LOX under supercritical and subcritical conditions was studied experimentally and theoretically. In experiments, the evaporation rate and surface temperature were measured for LOX strand vaporizing in helium environments at pressures ranging from 5 to 68 atmospheres. Gas sampling and chromatography analysis were also employed to profile the gas composition above the LOX surface for the purpose of model validation. A comprehensive theoretical model was formulated and solved numerically to simulate the evaporation process of LOX at high pressures. The model was based on the conservation equations of mass, momentum, energy, and species concentrations for a multicomponent system, with consideration of gravitational body force, solubility of ambient gases in liquid, and variable thermophysical properties. Good agreement between predictions and measured oxygen mole fraction profiles was obtained. The effect of pressure on the distribution of the Lewis number, as well as the effect of variable diffusion coefficient, were further examined to elucidate the high-pressure transport behavior exhibited in the LOX vaporization process.

2-Arylbenzo[b]furan derivatives as potent human lipoxygenase inhibitors.

PubMed

Lang, Li; Dong, Ningning; Wu, Deyan; Yao, Xue; Lu, Weiqiang; Zhang, Chen; Ouyang, Ping; Zhu, Jin; Tang, Yun; Wang, Wei; Li, Jian; Huang, Jin

2016-01-01

Human lipoxygenases (LOXs) have been emerging as effective therapeutic targets for inflammatory diseases. In this study, we found that four natural 2-arylbenzo[b]furan derivatives isolated from Artocarpus heterophyllus exhibited potent inhibitory activities against human LOXs, including moracin C (1), artoindonesianin B-1 (2), moracin D (3), moracin M (4). In our in vitro experiments, compound 1 was identified as the most potent LOX inhibitor and the moderate subtype selective inhibitor of 12-LOX. Compounds 1 and 2 act as competitive inhibitors of LOXs. Moreover, 1 significantly inhibits LTB4 production and chemotactic capacity of neutrophils, and is capable of protecting vascular barrier from plasma leakage in vivo. In addition, the preliminary structure-activity relationship analysis was performed based on the above four naturally occurring (1-4) and six additional synthetic 2-arylbenzo[b]furan derivatives. Taken together, these 2-arylbenzo[b]furan derivatives, as LOXs inhibitors, could represent valuable leads for the future development of therapeutic agents for inflammatory diseases.

LOX Gene transcript accumulation in olive (Olea europaea L.) fruits at different stages of maturation: relationship between volatile compounds, environmental factors, and technological treatments for oil extraction.

PubMed

Muzzalupo, Innocenzo; Macchione, Barbara; Bucci, Cristina; Stefanizzi, Francesca; Perri, Enzo; Chiappetta, Adriana; Tagarelli, Antonio; Sindona, Giovanni

2012-01-01

The quality of olive oil is influenced by genetic and environmental factors and by the maturation state of drupes, but it is equally affected by technological treatments of the process. This work investigates the possible correlation between olive LOX gene transcript accumulation, evaluated in fruits collected at different stages of maturation, and chemical biomarkers of its activity. During olive fruit ripening, the same genotype harvested from two different farms shows a positive linear trend between LOX relative transcript accumulation and the content of volatile compounds present in the olive oil aroma. Interestingly, a negative linear trend was observed between LOX relative transcript accumulation and the content of volatile compounds present in the olive pastes obtained from olive fruits with and without malaxation. The changes in the olive LOX transcript accumulation reveal its environmental regulation and suggest differential physiological functions for the LOXs.

Tumor-associated down-regulation of 15-lipoxygenase-1 is reversed by celecoxib in colorectal cancer.

PubMed

Heslin, Martin J; Hawkins, Ashley; Boedefeld, William; Arnoletti, J Pablo; Frolov, Andrey; Soong, Richie; Urist, Marshall M; Bland, Kirby I

2005-06-01

To evaluate the role of celecoxib on 15-lipoxygenase-1 (15-LOX-1) expression, protein levels, and rates of apoptosis in colorectal cancer cell lines. Also, to evaluate the expression of 15-LOX-1 in human normal mucosa, adenoma, and carcinoma with correlation to overall survival. The function of 15-LOX-1 is to maintain normal rates of apoptosis (programmed cell death). Decreased apoptosis is one mechanism of cancer growth and dissemination. It is our hypothesis that expression of 15-LOX-1 is reduced in human colorectal cancer (CRC) and the administration of celecoxib can reverse this process and induce apoptosis. Effect of celecoxib in cell culture: The effect of 40 micromol/L celecoxib was compared with untreated controls in tissue culture utilizing HT-29 and DLD-1 CRC cell lines. Expression of 15-LOX-1 protein was measured by immunoblot. Induction of apoptosis was evaluated by annexin V staining. All data are presented as mean +/- SEM, with significance defined as P < 0.05. 15-LOX-1 in human CRC: From February 1998 to January 2002, 126 patients underwent surgical resection of either colorectal adenomas (n = 24) or carcinomas (n = 102), or both (n = 25). Tissue was macrodissected, snap frozen, and stored at -80 degrees C. After tissue processing, RNA was extracted and gene expression of 15-LOX-1 was quantified utilizing ABI prism real-time quantitative RT-PCR. Significance evaluated by the Wilcoxon signed rank test. Effect of celecoxib in cell culture: After 72 hours of treatment with celecoxib, immunoblot demonstrated a 1.5- to 2-fold increase in 15-LOX-1 protein expression in HT-29 and DLD-1 cells, respectively. Celecoxib produced greater than a 2-fold increase in the rate of apoptosis compared with control cells in both cell lines (P < 0.05). 15-LOX-1 in human CRC: The mean age of the patients was 62 +/- 1 years; 78% were white and 48% were female. The mean size of the polyps and cancers were 3.0 +/- 0.4 and 5.0 +/- 0.1 cm, respectively. Expression of 15-LOX-1 relative to S9 was 30 in normal mucosa and significantly down-regulated to 11 in adenomas and 16 in carcinomas (P < 0.05). 15-LOX-1 gene expression is significantly reduced in both human colorectal adenomas and carcinomas and associated with decreased survival. Administration of celecoxib restores 15-LOX-1 protein expression and induces apoptosis. Down-regulation of 15-LOX-1 is an early event in the adenoma to carcinoma sequence, and reversal with celecoxib may represent one mechanism for chemoprevention of polyps or treatment of carcinomas.

Total hydrocarbon content (THC) testing in liquid oxygen (LOX) systems

NASA Astrophysics Data System (ADS)

Meneghelli, B. J.; Obregon, R. E.; Ross, H. R.; Hebert, B. J.; Sass, J. P.; Dirschka, G. E.

2015-12-01

The measured Total Hydrocarbon Content (THC) levels in liquid oxygen (LOX) systems at Stennis Space Center (SSC) have shown wide variations. Examples of these variations include the following: 1) differences between vendor-supplied THC values and those obtained using standard SSC analysis procedures; and 2) increasing THC values over time at an active SSC test stand in both storage and run vessels. A detailed analysis of LOX sampling techniques, analytical instrumentation, and sampling procedures will be presented. Additional data obtained on LOX system operations and LOX delivery trailer THC values during the past 12-24 months will also be discussed. Field test results showing THC levels and the distribution of the THC's in the test stand run tank, modified for THC analysis via dip tubes, will be presented.

Total Hydrocarbon Content (THC) Testing in Liquid Oxygen (LOX)

NASA Technical Reports Server (NTRS)

Meneghelli, B. J.; Obregon, R. E.; Ross, H. R.; Hebert, B. J.; Sass, J. P.; Dirschka, G. E.

2016-01-01

The measured Total Hydrocarbon Content (THC) levels in liquid oxygen (LOX) systems at Stennis Space Center (SSC) have shown wide variations. Examples of these variations include the following: 1) differences between vendor-supplied THC values and those obtained using standard SSC analysis procedures; and 2) increasing THC values over time at an active SSC test stand in both storage and run vessels. A detailed analysis of LOX sampling techniques, analytical instrumentation, and sampling procedures will be presented. Additional data obtained on LOX system operations and LOX delivery trailer THC values during the past 12-24 months will also be discussed. Field test results showing THC levels and the distribution of the THC's in the test stand run tank, modified for THC analysis via dip tubes, will be presented.

High-pressure LOX/hydrocarbon preburners and gas generators

NASA Technical Reports Server (NTRS)

Huebner, A. W.

1981-01-01

The objective of the program was to conduct a small scale hardware test program to establish the technology base required for LOX/hydrocarbon preburners and gas generators. The program consisted of six major tasks; Task I reviewed and assessed the performance prediction models and defined a subscale test program. Task II designed and fabricated this subscale hardware. Task III tested and analyzed the data from this hardware. Task IV analyzed the hot fire results and formulated a preliminary design for 40K preburner assemblies. Task V took the preliminary design and detailed and fabricated three 40K size preburner assemblies, one each fuel-rich LOX/CH, and LOX/RP-1 and one oxidizer rich LOX/CH4. Task VI delivered these preburner assemblies to MSFC for subsequent evaluation.

Structure and interaction with phospholipids of a prokaryotic lipoxygenase from Pseudomonas aeruginosa

PubMed Central

Garreta, Albert; Val-Moraes, Silvana P.; García-Fernández, Queralt; Busquets, Montserrat; Juan, Carlos; Oliver, Antonio; Ortiz, Antonio; Gaffney, Betty J.; Fita, Ignacio; Manresa, Àngels; Carpena, Xavi

2013-01-01

Lipoxygenases (LOXs), which are essential in eukaryotes, have no confirmed function in prokaryotes that are devoid of polyunsaturated fatty acids. The structure of a secretable LOX from Pseudomonas aeruginosa (Pa_LOX), the first available from a prokaryote, presents significant differences with respect to eukaryotic LOXs, including a cluster of helices acting as a lid to the active center. The mobility of the lid and the structural variability of the N-terminal region of Pa_LOX was confirmed by comparing 2 crystal forms. The binding pocket contains a phosphatidylethanolamine phospholipid with branches of 18 (sn-1) and 14/16 (sn-2) carbon atoms in length. Carbon atoms from the sn-1 chain approach the catalytic iron in a manner that sheds light on how the enzymatic reaction might proceed. The findings in these studies suggest that Pa_LOX has the capacity to extract and modify unsaturated phospholipids from eukaryotic membranes, allowing this LOX to play a role in the interaction of P. aeruginosa with host cells.—Garreta, A., Val-Moraes, S. P., García-Fernández, Q., Montserrat Busquets, C. J., Oliver, A., Ortiz, A., Gaffney, B. J., Fita, I., Manresa, A., Carpena, X. Structure and interaction with phospholipids of a prokaryotic lipoxygenase from Pseudomonas aeruginosa. PMID:23985801

Inhibition of Lysyl Oxidases Impairs Migration and Angiogenic Properties of Tumor-Associated Pericytes.

PubMed

Ribeiro, Aline Lopes; Kaid, Carolini; Silva, Patrícia B G; Cortez, Beatriz A; Okamoto, Oswaldo Keith

2017-01-01

Pericytes are important cellular components of the tumor microenviroment with established roles in angiogenesis and metastasis. These two cancer hallmarks are modulated by enzymes of the LOX family, but thus far, information about LOX relevance in tumor-associated pericytes is lacking. Here, we performed a comparative characterization of normal and tumoral pericytes and report for the first time the modulatory effects of LOX enzymes on activated pericyte properties. Tumoral pericytes isolated from childhood ependymoma and neuroblastoma specimens displayed angiogenic properties in vitro and expressed typical markers, including CD146, NG2, and PDGFR β . Expression of all LOX family members could be detected in both normal and tumor-associated pericytes. In most pericyte samples, LOXL3 was the family member displaying the highest transcript levels. Inhibition of LOX/LOXL activity with the inhibitor β -aminopropionitrile ( β APN) significantly reduced migration of pericytes, while proliferation rates were kept unaltered. Formation of tube-like structures in vitro by pericytes was also significantly impaired upon inhibition of LOX/LOXL activity with β APN, which induced more prominent effects in tumor-associated pericytes. These findings reveal a novel involvement of the LOX family of enzymes in migration and angiogenic properties of pericytes, with implications in tumor development and in therapeutic targeting tumor microenvironment constituents.

Inhibition of Lysyl Oxidases Impairs Migration and Angiogenic Properties of Tumor-Associated Pericytes

PubMed Central

Kaid, Carolini; Silva, Patrícia B. G.; Cortez, Beatriz A.

2017-01-01

Pericytes are important cellular components of the tumor microenviroment with established roles in angiogenesis and metastasis. These two cancer hallmarks are modulated by enzymes of the LOX family, but thus far, information about LOX relevance in tumor-associated pericytes is lacking. Here, we performed a comparative characterization of normal and tumoral pericytes and report for the first time the modulatory effects of LOX enzymes on activated pericyte properties. Tumoral pericytes isolated from childhood ependymoma and neuroblastoma specimens displayed angiogenic properties in vitro and expressed typical markers, including CD146, NG2, and PDGFRβ. Expression of all LOX family members could be detected in both normal and tumor-associated pericytes. In most pericyte samples, LOXL3 was the family member displaying the highest transcript levels. Inhibition of LOX/LOXL activity with the inhibitor β-aminopropionitrile (βAPN) significantly reduced migration of pericytes, while proliferation rates were kept unaltered. Formation of tube-like structures in vitro by pericytes was also significantly impaired upon inhibition of LOX/LOXL activity with βAPN, which induced more prominent effects in tumor-associated pericytes. These findings reveal a novel involvement of the LOX family of enzymes in migration and angiogenic properties of pericytes, with implications in tumor development and in therapeutic targeting tumor microenvironment constituents. PMID:28553358

Serum amyloid A stimulates macrophage foam cell formation via lectin-like oxidized low-density lipoprotein receptor 1 upregulation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lee, Ha Young, E-mail: hayoung@skku.edu; Mitochondria Hub Regulation Center, Dong-A University, Busan 602-714; Kim, Sang Doo

2013-03-29

Highlights: ► SAA induced macrophage foam cell formation. ► SAA stimulated upregulation of lectin-like oxidized low-density lipoprotein receptor 1 (LOX1). ► SAA-induced LOX1 expression and foam cell formation is mediated by JNK/NF-κB signaling. ► HDL-conjugated SAA also stimulates foam cell formation via LOX1 upregulation. ► The finding reveals a novel mechanism of action of SAA in the pathogenesis of atherosclerosis. -- Abstract: Elevated levels of serum amyloid A (SAA) is a risk factor for cardiovascular diseases, however, the role of SAA in the pathophysiology of atherosclerosis remains unclear. Here we show that SAA induced macrophage foam cell formation. SAA-stimulated foammore » cell formation was mediated by c-jun N-terminal kinase (JNK) signaling. Moreover, both SAA and SAA-conjugated high density lipoprotein stimulated the expression of the important scavenger receptor lectin-like oxidized low-density lipoprotein receptor 1 (LOX1) via nuclear factor-κB (NF-κB). A LOX1 antagonist carrageenan significantly blocked SAA-induced foam cell formation, indicating that SAA promotes foam cell formation via LOX1 expression. Our findings therefore suggest that SAA stimulates foam cell formation via LOX1 induction, and thus likely contributes to atherogenesis.« less

Protective Effects of Let-7a and Let-7b on Oxidized Low-Density Lipoprotein Induced Endothelial Cell Injuries

PubMed Central

Bao, Mei-hua; Zhang, Yi-wen; Lou, Xiao-ya; Cheng, Yu; Zhou, Hong-hao

2014-01-01

Lectin-like low-density lipoprotein receptor 1 (LOX-1) is a receptor for oxidized low density lipoprotein (oxLDL) in endothelial cells. The activation of LOX-1 by oxLDL stimulates the apoptosis and dysfunction of endothelial cells, and contributes to atherogenesis. However, the regulatory factors for LOX-1 are still unclear. MicroRNAs are small, endogenous, non-coding RNAs that regulate gene expressions at a post-transcriptional level. The let-7 family is the second microRNA been discovered, which plays important roles in cardiovascular diseases. Let-7a and let-7b were predicted to target LOX-1 3′-UTR and be highly expressed in endothelial cells. The present study demonstrated that LOX-1 was a target of let-7a and let-7b. They inhibited the expression of LOX-1 by targeting the positions of 310-316 in LOX-1 3′-UTR. Over-expression of let-7a and let-7b inhibited the oxLDL-induced endothelial cell apoptosis, NO deficiency, ROS over-production, LOX-1 upregulation and endothelial nitric oxide synthase (eNOS) downregulation. Moreover, we found that oxLDL treatment induced p38MAPK phosphorylation, NF-κB nuclear translocation, IκB degradation and PKB dephosphorylation. Let-7a or let-7b over-expression attenuated these alterations significantly. The present study may provide a new insight into the protective properties of let-7a and let-7b in preventing the endothelial dysfunction associated with cardiovascular disease, such as atherosclerosis. PMID:25247304

LOX/Hydrocarbon Combustion Instability Investigation

NASA Technical Reports Server (NTRS)

Jensen, R. J.; Dodson, H. C.; Claflin, S. E.

1989-01-01

The LOX/Hydrocarbon Combustion Instability Investigation Program was structured to determine if the use of light hydrocarbon combustion fuels with liquid oxygen (LOX) produces combustion performance and stability behavior similar to the LOX/hydrogen propellant combination. In particular methane was investigated to determine if that fuel can be rated for combustion instability using the same techniques as previously used for LOX/hydrogen. These techniques included fuel temperature ramping and stability bomb tests. The hot fire program probed the combustion behavior of methane from ambient to subambient temperatures. Very interesting results were obtained from this program that have potential importance to future LOX/methane development programs. A very thorough and carefully reasoned documentation of the experimental data obtained is contained. The hot fire test logic and the associated tests are discussed. Subscale performance and stability rating testing was accomplished using 40,000 lb. thrust class hardware. Stability rating tests used both bombs and fuel temperature ramping techniques. The test program was successful in generating data for the evaluation of the methane stability characteristics relative to hydrogen and to anchor stability models. Data correlations, performance analysis, stability analyses, and key stability margin enhancement parameters are discussed.

Performance of a shaft seal system for the LE-7 rocket engine oxidizer turbopump

NASA Astrophysics Data System (ADS)

Oike, Mamoru; Nosaka, Masataka; Kikuchi, Masataka; Watanabe, Yoshiaki

An experimental study on a rotating-shaft seal system for a high-pressure liquid oxygen (LOX) turbopump has been conducted to develop the LE-7 engine for the Japanese H-II launch vehicle. The LOX turbopump rotating-shaft seal system, which prevents LOX (4.9 MPa) and the high-pressure turbine-drive gas (16.6 MPa, 970 K) from mixing during high-speed operations (18,000 to 20,000 rpm), consists of the following seals: an LOX seal comprising a floating-ring and a wear-ring, a turbine gas seal comprising two floating-rings, and a helium purge seal comprising two segmented circumferential seal-rings. This report describes experimental and observational results concerning the rotating-shaft seal system obtained in the LOX turbopump operations and the seal tests. Based on comparisons between the measurements and the analytical results, sealing characteristics of the seal system are discussed. Inspections of the sealing surfaces after the engine firing tests demonstrated that the LOX turbopump rotating-shaft seal system has sufficient durability for use in the LE-7 engine for the H-II launch vehicle.

Oxidized LDL binding to LOX-1 upregulates VEGF expression in cultured bovine chondrocytes through activation of PPAR-{gamma}

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kanata, Sohya; Akagi, Masao; Nishimura, Shunji

It has been reported that vascular endothelial growth factor (VEGF) and its receptors play an important role in the destruction of articular cartilage in osteoarthritis through increased production of matrix metalloproteinases. We investigated whether the oxidized low-density lipoprotein (ox-LDL) binding to lectin-like ox-LDL receptor-1 (LOX-1) upregulates VEGF expression in cultured bovine articular chondrocytes (BACs). Ox-LDL markedly increased VEGF mRNA expression and protein release in time- and dose-dependent manners, which was significantly suppressed by anti-LOX-1 antibody pretreatment. Activation of peroxisome proliferator-activated receptor (PPAR)-{gamma} was evident in BACs with ox-LDL addition and was attenuated by anti-LOX-1 antibody. The specific PPAR-{gamma} inhibitor GW9662more » suppressed ox-LDL-induced VEGF expression. These results suggest that the ox-LDL/LOX-1 system upregulates VEGF expression in articular cartilage, at least in part, through activation of PPAR-{gamma} and supports the hypothesis that ox-LDL is involved in cartilage degradation via LOX-1.« less

Expression of lectin-like oxidized LDL receptor-1 in smooth muscle cells after vascular injury

DOE Office of Scientific and Technical Information (OSTI.GOV)

Eto, Hideyuki; Miyata, Masaaki; Kume, Noriaki

Lectin-like oxidized LDL receptor-1 (LOX-1) is an oxidized LDL receptor, and its role in restenosis after angioplasty remains unknown. We used a balloon-injury model of rabbit aorta, and reverse transcription-polymerase chain reaction revealed that LOX-1 mRNA expression was modest in the non-injured aorta, reached a peak level 2 days after injury, and remained elevated until 24 weeks after injury. Immunohistochemistry and in situ hybridization showed that LOX-1 was not detected in the media of non-injured aorta but expressed in both medial and neointimal smooth muscle cells (SMC) at 2 and 24 weeks after injury. Low concentrations of ox-LDL (10 {mu}g/mL)more » stimulated the cultured SMC proliferation, which was inhibited by antisense oligonucleotides of LOX-1 mRNA. Double immunofluorescense staining showed the colocalization of LOX-1 and proliferating cell nuclear antigen in human restenotic lesion. These results suggest that LOX-1 mediates ox-LDL-induced SMC proliferation and plays a role in neointimal formation after vascular injury.« less

Expression of lectin-like oxidized LDL receptor-1 in smooth muscle cells after vascular injury.

PubMed

Eto, Hideyuki; Miyata, Masaaki; Kume, Noriaki; Minami, Manabu; Itabe, Hiroyuki; Orihara, Koji; Hamasaki, Shuichi; Biro, Sadatoshi; Otsuji, Yutaka; Kita, Toru; Tei, Chuwa

2006-03-10

Lectin-like oxidized LDL receptor-1 (LOX-1) is an oxidized LDL receptor, and its role in restenosis after angioplasty remains unknown. We used a balloon-injury model of rabbit aorta, and reverse transcription-polymerase chain reaction revealed that LOX-1 mRNA expression was modest in the non-injured aorta, reached a peak level 2 days after injury, and remained elevated until 24 weeks after injury. Immunohistochemistry and in situ hybridization showed that LOX-1 was not detected in the media of non-injured aorta but expressed in both medial and neointimal smooth muscle cells (SMC) at 2 and 24 weeks after injury. Low concentrations of ox-LDL (10 microg/mL) stimulated the cultured SMC proliferation, which was inhibited by antisense oligonucleotides of LOX-1 mRNA. Double immunofluorescence staining showed the colocalization of LOX-1 and proliferating cell nuclear antigen in human restenotic lesion. These results suggest that LOX-1 mediates ox-LDL-induced SMC proliferation and plays a role in neointimal formation after vascular injury.

LOX/hydrocarbon rocket engine analytical design methodology development and validation. Volume 1: Executive summary and technical narrative

NASA Technical Reports Server (NTRS)

Pieper, Jerry L.; Walker, Richard E.

1993-01-01

During the past three decades, an enormous amount of resources were expended in the design and development of Liquid Oxygen/Hydrocarbon and Hydrogen (LOX/HC and LOX/H2) rocket engines. A significant portion of these resources were used to develop and demonstrate the performance and combustion stability for each new engine. During these efforts, many analytical and empirical models were developed that characterize design parameters and combustion processes that influence performance and stability. Many of these models are suitable as design tools, but they have not been assembled into an industry-wide usable analytical design methodology. The objective of this program was to assemble existing performance and combustion stability models into a usable methodology capable of producing high performing and stable LOX/hydrocarbon and LOX/hydrogen propellant booster engines.

Flow-induced Vibration of SSME Main Injector Liquid-oxygen Posts

NASA Technical Reports Server (NTRS)

Chen, S. S.; Jendrzejczyk, J. A.; Wambsganss, M. W.

1985-01-01

The liquid-oxygen (LOX) posts are exposed to hot hydrogen flowing over the tubes on its way to the combustion chamber. Fatigue cracking of some LOX posts was observed after test firing of the SSMEs. A current design modification consists of attaching impingement shields to the LOX posts in the outer row. The modification improved the vibration/fatigue problem of the LOX posts, but resulted in an increased pressure drop that ultimately shortened the life expectancy of other components. A fundamental study of vibration of the LOX posts was initiated to understand the flow-induced vibration problem and to develop techniques to avoid detrimental vibrational effects with the overall objective of improving engine life. This effort, including an assessment of the problem, scoping calculation and experiment, and a work plan for an integrated theoretical/experimental study of the problem is summarized.

A revolutionary lunar space transportation system architecture using extraterrestrial LOX-augmented NTR propulsion

NASA Astrophysics Data System (ADS)

Borowski, Stanley K.; Corban, Robert R.; Culver, Donald W.; Bulman, Melvin J.; McIlwain, Mel C.

1994-08-01

The concept of a liquid oxygen (LOX)-augmented nuclear thermal rocket (NTR) engine is introduced, and its potential for revolutionizing lunar space transportation system (LTS) performance using extraterrestrial 'lunar-derived' liquid oxygen (LUNOX) is outlined. The LOX-augmented NTR (LANTR) represents the marriage of conventional liquid hydrogen (LH2)-cooled NTR and airbreathing engine technologies. The large divergent section of the NTR nozzle functions as an 'afterburner' into which oxygen is injected and supersonically combusted with nuclear preheated hydrogen emerging from the NTR's choked sonic throat: 'scramjet propulsion in reverse.' By varying the oxygen-to-fuel mixture ratio (MR), the LANTR concept can provide variable thrust and specific impulse (Isp) capability with a LH2-cooled NTR operating at relatively constant power output. For example, at a MR = 3, the thrust per engine can be increased by a factor of 2.75 while the Isp decreases by only 30 percent. With this thrust augmentation option, smaller, 'easier to develop' NTR's become more acceptable from a mission performance standpoint (e.g., earth escape gravity losses are reduced and perigee propulsion requirements are eliminated). Hydrogen mass and volume is also reduced resulting in smaller space vehicles. An evolutionary NTR-based lunar architecture requiring only Shuttle C and/or 'in-line' shuttle-derived launch vehicles (SDV's) would operate initially in an 'expandable mode' with NTR lunar transfer vehicles (LTV's) delivering 80 percent more payload on piloted missions than their LOX/LH2 chemical propulsion counterparts. With the establishment of LUNOX production facilities on the lunar surface and 'fuel/oxidizer' depot in low lunar orbit (LLO), monopropellant NTR's would be outfitted with an oxygen propellant module, feed system, and afterburner nozzle for 'bipropellant' operation. The LANTR cislunar LTV now transitions to a reusable mode with smaller vehicle and payload doubling benefits on each piloted round trip mission. As the initial lunar outposts grow to centralized bases and settlements with a substantial permanent human presence, a LANTR-powered shuttle capable of 36 to 24 hour 'one-way' trip times to the moon and back becomes possible with initial mass in low earth orbit (IMLEO) requirements of approximately 160 to 240 metric tons, respectively.

A Revolutionary Lunar Space Transportation System Architecture Using Extraterrestrial Lox-augmented NTR Propulsion

NASA Technical Reports Server (NTRS)

Borowski, Stanley K.; Corban, Robert R.; Culver, Donald W.; Bulman, Melvin J.; Mcilwain, Mel C.

1994-01-01

The concept of a liquid oxygen (LOX)-augmented nuclear thermal rocket (NTR) engine is introduced, and its potential for revolutionizing lunar space transportation system (LTS) performance using extraterrestrial 'lunar-derived' liquid oxygen (LUNOX) is outlined. The LOX-augmented NTR (LANTR) represents the marriage of conventional liquid hydrogen (LH2)-cooled NTR and airbreathing engine technologies. The large divergent section of the NTR nozzle functions as an 'afterburner' into which oxygen is injected and supersonically combusted with nuclear preheated hydrogen emerging from the NTR's choked sonic throat: 'scramjet propulsion in reverse.' By varying the oxygen-to-fuel mixture ratio (MR), the LANTR concept can provide variable thrust and specific impulse (Isp) capability with a LH2-cooled NTR operating at relatively constant power output. For example, at a MR = 3, the thrust per engine can be increased by a factor of 2.75 while the Isp decreases by only 30 percent. With this thrust augmentation option, smaller, 'easier to develop' NTR's become more acceptable from a mission performance standpoint (e.g., earth escape gravity losses are reduced and perigee propulsion requirements are eliminated). Hydrogen mass and volume is also reduced resulting in smaller space vehicles. An evolutionary NTR-based lunar architecture requiring only Shuttle C and/or 'in-line' shuttle-derived launch vehicles (SDV's) would operate initially in an 'expandable mode' with NTR lunar transfer vehicles (LTV's) delivering 80 percent more payload on piloted missions than their LOX/LH2 chemical propulsion counterparts. With the establishment of LUNOX production facilities on the lunar surface and 'fuel/oxidizer' depot in low lunar orbit (LLO), monopropellant NTR's would be outfitted with an oxygen propellant module, feed system, and afterburner nozzle for 'bipropellant' operation. The LANTR cislunar LTV now transitions to a reusable mode with smaller vehicle and payload doubling benefits on each piloted round trip mission. As the initial lunar outposts grow to centralized bases and settlements with a substantial permanent human presence, a LANTR-powered shuttle capable of 36 to 24 hour 'one-way' trip times to the moon and back becomes possible with initial mass in low earth orbit (IMLEO) requirements of approximately 160 to 240 metric tons, respectively.

Kinetic investigation of human 5-lipoxygenase with arachidonic acid.

PubMed

Mittal, Monica; Kumar, Ramakrishnan B; Balagunaseelan, Navisraj; Hamberg, Mats; Jegerschöld, Caroline; Rådmark, Olof; Haeggström, Jesper Z; Rinaldo-Matthis, Agnes

2016-08-01

Human 5-lipoxygenase (5-LOX) is responsible for the formation of leukotriene (LT)A4, a pivotal intermediate in the biosynthesis of the leukotrienes, a family of proinflammatory lipid mediators. 5-LOX has thus gained attention as a potential drug target. However, details of the kinetic mechanism of 5-LOX are still obscure. In this Letter, we investigated the kinetic isotope effect (KIE) of 5-LOX with its physiological substrate, arachidonic acid (AA). The observed KIE is 20±4 on kcat and 17±2 on kcat/KM at 25°C indicating a non-classical reaction mechanism. The observed rates show slight temperature dependence at ambient temperatures ranging from 4 to 35°C. Also, we observed low Arrhenius prefactor ratio (AH/AD=0.21) and a small change in activation energy (Ea(D)-Ea(H)=3.6J/mol) which suggests that 5-LOX catalysis involves tunneling as a mechanism of H-transfer. The measured KIE for 5-LOX involves a change in regioselectivity in response to deuteration at position C7, resulting in H-abstraction form C10 and formation of 8-HETE. The viscosity experiments influence the (H)kcat, but not (D)kcat. However the overall kcat/KM is not affected for labeled or unlabeled AA, suggesting that either the product release or conformational rearrangement might be involved in dictating kinetics of 5-LOX at saturating conditions. Investigation of available crystal structures suggests the role of active site residues (F421, Q363 and L368) in regulating the donor-acceptor distances, thus affecting H-transfer as well as regiospecificity. In summary, our study shows that that the H-abstraction is the rate limiting step for 5-LOX and that the observed KIE of 5-LOX is masked by a change in regioselectivity. Copyright © 2016 Elsevier Ltd. All rights reserved.

Endothelial LOX-1 activation differentially regulates arterial thrombus formation depending on oxLDL levels: role of the Oct-1/SIRT1 and ERK1/2 pathways.

PubMed

Akhmedov, Alexander; Camici, Giovanni G; Reiner, Martin F; Bonetti, Nicole R; Costantino, Sarah; Holy, Erik W; Spescha, Remo D; Stivala, Simona; Schaub Clerigué, Ariane; Speer, Thimoteus; Breitenstein, Alexander; Manz, Jasmin; Lohmann, Christine; Paneni, Francesco; Beer, Juerg-Hans; Lüscher, Thomas F

2017-04-01

The lectin-like oxLDL receptor-1 (LOX-1) promotes endothelial uptake of oxidized low-density lipoprotein (oxLDL) and plays an important role in atherosclerosis and acute coronary syndromes (ACS). However, its role in arterial thrombus formation remains unknown. We investigated whether LOX-1 plays a role in arterial thrombus formation in vivo at different levels of oxLDL using endothelial-specific LOX-1 transgenic mice (LOX-1TG) and a photochemical injury thrombosis model of the carotid artery. In mice fed a normal chow diet, time to arterial occlusion was unexpectedly prolonged in LOX-1TG as compared to WT. In line with this, tissue factor (TF) expression and activity in carotid arteries of LOX-1TG mice were reduced by half. This effect was mediated by activation of octamer transcription factor 1 (Oct-1) leading to upregulation of the mammalian deacetylase silent information regulator-two 1 (SIRT1) via binding to its promoter and subsequent inhibition of NF-κB signaling. In contrast, intravenous injection of oxLDL as well as high cholesterol diet for 6 weeks led to a switch from the Oct-1/SIRT1 signal transduction pathway to the ERK1/2 pathway and in turn to an enhanced thrombotic response with shortened occlusion time. Thus, LOX-1 differentially regulates thrombus formation in vivo depending on the degree of activation by oxLDL. At low oxLDL levels LOX-1 activates the protective Oct-1/SIRT1 pathway, while at higher levels of the lipoprotein switches to the thrombogenic ERK1/2 pathway. These findings may be important for arterial thrombus formation in ACS and suggest that SIRT1 may represent a novel therapeutic target in this context. Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2017. For permissions, please email: journals.permissions@oup.com.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jang, Insook; Park, Sujin; Cho, Jin Won

12/15-lipoxygenase (12/15-LOX) is involved in organelle homeostasis by degrading mitochondria in maturing red blood cells and by eliminating excess peroxisomes in liver. Furthermore, 12/15-LOX contributes to diseases by exacerbating oxidative stress-related injury, notably in stroke. Nonetheless, it is unclear what the consequences are of abolishing 12/15-LOX activity. Mice in which the alox15 gene has been ablated do not show an obvious phenotype, and LOX enzyme inhibition is not overtly detrimental. We show here that liver histology is also unremarkable. However, electron microscopy demonstrated that 12/15-LOX knockout surprisingly leads to increased macroautophagy in the liver. Not only macroautophagy but also mitophagymore » and pexophagy were increased in hepatocytes, which otherwise showed unaltered fine structure and organelle morphology. These findings were substantiated by immunofluorescence showing significantly increased number of LC3 puncta and by Western blotting demonstrating a significant increase for LC3-II protein in both liver and brain homogenates of 12/15-LOX knockout mice. Inhibition of 12/15-LOX activity by treatment with four structurally different inhibitors had similar effects in cultured HepG2 hepatoma cells and SH-SY5Y neuroblastoma cells with significantly increased autophagy discernable already after 2 hours. Hence, our study reveals a link between ablation or inhibition of 12/15-LOX and stimulation of macroautophagy. The enhanced macroautophagy may be related to the known tissue-protective effects of LOX ablation or inhibition under various diseased conditions caused by oxidative stress and ischemia. This could provide an important cleaning mechanism of cells and tissues to prevent accumulation of damaged mitochondria and other cellular components. - Highlights: • A relationship between lipoxygenases and autophagy is disclosed. • 12/15-lipoxygenase knockout increases autophagy in mice liver and brain. • Lipoxygenase inhibition boosts autophagy in human hepatoma and neuroblastoma cells. • Lipoxygenase knockout or inhibition triggers selective autophagy.« less

Tumor Tension Induces Persistent Inflammation and Promotes Breast Cancer Aggression

DTIC Science & Technology

2015-10-01

assessing collagen crosslinking, fibrillar collagen I deposition, and macrophage infiltration in a pilot study using MMTV-PyMT; Col1a1 -tTA;TetO_mLOX model...where LOX was overexpressed in Col1a1 + activated fibroblasts (Figure 4B-C). The second model is an inducible LOX knockout model: MMTV-PyMT; Col1a1 ...cells were isolated and analyzed using flow cytometry. (B) MMTV-PyMT; Col1a1 -tTA;TetO_mLOX mice have been taken off DOX treatment at 6 weeks of age

Exploring the Functions of 9-Lipoxygenase (DkLOX3) in Ultrastructural Changes and Hormonal Stress Response during Persimmon Fruit Storage

PubMed Central

Meng, Kun; Hou, Yali; Han, Ye; Ban, Qiuyan; He, Yiheng; Suo, Jiangtao; Rao, Jingping

2017-01-01

Lipoxygenase (LOX) initiates the hydroperoxidation of polyunsaturated fatty acids and is involved in multiple physiological processes. In this study, investigation of various microscopic techniques showed that the fruit peel cellular microstructure of the two persimmon cultivars differed after 12 days of storage, resulting in fruit weight loss and an increased number and depth of microcracks. Analysis of subcellular localization revealed that greater amounts of DkLOX3-immunolabelled gold particles accumulated in “Fupingjianshi” than in “Ganmaokui” during storage. In addition, the expression of DkLOX3 was positively up-regulated by abscisic acid (ABA), concomitant with the promotion of ethylene synthesis and loss of firmness, and was suppressed by salicylic acid (SA), concomitant with the maintenance of fruit firmness, inhibition of ethylene production and weight loss. In particular, the expression of DkLOX3 differed from the ethylene trajectory after methyl jasmonate (MeJA) treatment. Furthermore, we isolated a 1105 bp 5′ flanking region of DkLOX3 and the activity of promoter deletion derivatives was induced through various hormonal treatments. Promoter sequence cis-regulatory elements were analysed, and two conserved hormone-responsive elements were found to be essential for responsiveness to hormonal stress. Overall, these results will provide us with new clues for exploring the functions of DkLOX3 in fruit ripening and hormonal stress response. PMID:28294971

Integrated Pressure-Fed Liquid Oxygen / Methane Propulsion Systems - Morpheus Experience, MARE, and Future Applications

NASA Technical Reports Server (NTRS)

Hurlbert, Eric; Morehead, Robert; Melcher, John C.; Atwell, Matt

2016-01-01

An integrated liquid oxygen (LOx) and methane propulsion system where common propellants are fed to the reaction control system and main engines offers advantages in performance, simplicity, reliability, and reusability. LOx/Methane provides new capabilities to use propellants that are manufactured on the Mars surface for ascent return and to integrate with power and life support systems. The clean burning, non-toxic, high vapor pressure propellants provide significant advantages for reliable ignition in a space vacuum, and for reliable safing or purging of a space-based vehicle. The NASA Advanced Exploration Systems (AES) Morpheus lander demonstrated many of these key attributes as it completed over 65 tests including 15 flights through 2014. Morpheus is a prototype of LOx/Methane propellant lander vehicle with a fully integrated propulsion system. The Morpheus lander flight demonstrations led to the proposal to use LOx/Methane for a Discovery class mission, named Moon Aging Regolith Experiment (MARE) to land an in-situ science payload for Southwest Research Institute on the Lunar surface. Lox/Methane is extensible to human spacecraft for many transportation elements of a Mars architecture. This paper discusses LOx/Methane propulsion systems in regards to trade studies, the Morpheus project experience, the MARE NAVIS (NASA Autonomous Vehicle for In-situ Science) lander, and future possible applications. The paper also discusses technology research and development needs for Lox/Methane propulsion systems.

Up-Regulated Expression of AOS-LOXa and Increased Eicosanoid Synthesis in Response to Coral Wounding

PubMed Central

Lõhelaid, Helike; Teder, Tarvi; Tõldsepp, Kadri; Ekins, Merrick; Samel, Nigulas

2014-01-01

In octocorals, a catalase–like allene oxide synthase (AOS) and an 8R-lipoxygenase (LOX) gene are fused together encoding for a single AOS-LOX fusion protein. Although the AOS-LOX pathway is central to the arachidonate metabolism in corals, its biological function in coral homeostasis is unclear. Using an acute incision wound model in the soft coral Capnella imbricata, we here test whether LOX pathway, similar to its role in plants, can contribute to the coral damage response and regeneration. Analysis of metabolites formed from exogenous arachidonate before and after fixed time intervals following wounding indicated a significant increase in AOS-LOX activity in response to mechanical injury. Two AOS-LOX isoforms, AOS-LOXa and AOS-LOXb, were cloned and expressed in bacterial expression system as active fusion proteins. Transcription levels of corresponding genes were measured in normal and stressed coral by qPCR. After wounding, AOS-LOXa was markedly up-regulated in both, the tissue adjacent to the incision and distal parts of a coral colony (with the maximum reached at 1 h and 6 h post wounding, respectively), while AOS-LOXb was stable. According to mRNA expression analysis, combined with detection of eicosanoid product formation for the first time, the AOS-LOX was identified as an early stress response gene which is induced by mechanical injury in coral. PMID:24551239

Developing functional musculoskeletal tissues through hypoxia and lysyl oxidase-induced collagen cross-linking

PubMed Central

Makris, Eleftherios A.; Responte, Donald J.; Hu, Jerry C.; Athanasiou, Kyriacos A.

2014-01-01

The inability to recapitulate native tissue biomechanics, especially tensile properties, hinders progress in regenerative medicine. To address this problem, strategies have focused on enhancing collagen production. However, manipulating collagen cross-links, ubiquitous throughout all tissues and conferring mechanical integrity, has been underinvestigated. A series of studies examined the effects of lysyl oxidase (LOX), the enzyme responsible for the formation of collagen cross-links. Hypoxia-induced endogenous LOX was applied in multiple musculoskeletal tissues (i.e., cartilage, meniscus, tendons, ligaments). Results of these studies showed that both native and engineered tissues are enhanced by invoking a mechanism of hypoxia-induced pyridinoline (PYR) cross-links via intermediaries like LOX. Hypoxia was shown to enhance PYR cross-linking 1.4- to 6.4-fold and, concomitantly, to increase the tensile properties of collagen-rich tissues 1.3- to 2.2-fold. Direct administration of exogenous LOX was applied in native cartilage and neocartilage generated using a scaffold-free, self-assembling process of primary chondrocytes. Exogenous LOX was found to enhance native tissue tensile properties 1.9-fold. LOX concentration- and time-dependent increases in PYR content (∼16-fold compared with controls) and tensile properties (approximately fivefold compared with controls) of neocartilage were also detected, resulting in properties on par with native tissue. Finally, in vivo subcutaneous implantation of LOX-treated neocartilage in nude mice promoted further maturation of the neotissue, enhancing tensile and PYR content approximately threefold and 14-fold, respectively, compared with in vitro controls. Collectively, these results provide the first report, to our knowledge, of endogenous (hypoxia-induced) and exogenous LOX applications for promoting collagen cross-linking and improving the tensile properties of a spectrum of native and engineered tissues both in vitro and in vivo. PMID:25349395

The role of the lectin-like oxLDL receptor (LOX-1) in traffic-generated air pollution exposure-mediated alteration of the brain microvasculature in Apolipoprotein (Apo) E knockout mice.

PubMed

Lucero, JoAnn; Suwannasual, Usa; Herbert, Lindsay M; McDonald, Jacob D; Lund, Amie K

2017-05-01

Recent studies have shown a strong correlation between air pollution-exposure and detrimental outcomes in the central nervous system, including alterations in blood brain barrier (BBB) integrity, neuroinflammation, and neurodegeneration. However, the mechanisms mediating these pathologies have not yet been fully elucidated. We have previously reported that exposure to traffic-generated air pollution results in increased circulating oxidized low-density lipoprotein (oxLDL), associated with alterations in BBB integrity, in atherosclerotic Apolipoprotein E null (ApoE -/- ) mice. Thus, we investigated the role of the lectin-like oxLDL receptor (LOX)-1 in mediating these deleterious effects in ApoE -/- mice exposed to a mixture of gasoline and diesel engine exhaust (MVE: 100 PM µg/m 3 ) for 6 h/d, 7d/week, for 30 d by inhalation. Concurrent with exposures, a subset of mice were treated with neutralizing antibodies to LOX-1 (LOX-1 Ab) i.p., or IgG (control) i.p., every other day during exposures. Resulting brain microvascular integrity, tight junction (TJ) protein expression, matrix metalloproteinase (MMP)-9/-2 activity, ROS, and markers of cellular adhesion and monocyte/macrophage sequestration were assessed. MVE-exposure resulted in decreased BBB integrity and alterations in microvascular TJ protein expression, associated with increased LOX-1 expression, MMP-9/-2 activities, and lipid peroxidation, each of which was attenuated with LOX-1 Ab treatment. Furthermore, MVE-exposure induced cerebral microvascular ROS and adhesion molecules, expression of which was not normalized through LOX-1 Ab-treatment. Such findings suggest that alterations in brain microvascular structure and integrity observed with MVE-exposure may be mediated, at least in part, via LOX-1 signaling.

First Selective 12-LOX Inhibitor, ML355, Impairs Thrombus Formation and Vessel Occlusion In Vivo With Minimal Effects on Hemostasis.

PubMed

Adili, Reheman; Tourdot, Benjamin E; Mast, Katherine; Yeung, Jennifer; Freedman, John C; Green, Abigail; Luci, Diane K; Jadhav, Ajit; Simeonov, Anton; Maloney, David J; Holman, Theodore R; Holinstat, Michael

2017-10-01

Adequate platelet reactivity is required for maintaining hemostasis. However, excessive platelet reactivity can also lead to the formation of occlusive thrombi. Platelet 12(S)-lipoxygenase (12-LOX), an oxygenase highly expressed in the platelet, has been demonstrated to regulate platelet function and thrombosis ex vivo, supporting a key role for 12-LOX in the regulation of in vivo thrombosis. However, the ability to pharmacologically target 12-LOX in vivo has not been established to date. Here, we studied the effect of the first highly selective 12-LOX inhibitor, ML355, on in vivo thrombosis and hemostasis. ML355 dose-dependently inhibited human platelet aggregation and 12-LOX oxylipin production, as confirmed by mass spectrometry. Interestingly, the antiplatelet effects of ML355 were reversed after exposure to high concentrations of thrombin in vitro. Ex vivo flow chamber assays confirmed that human platelet adhesion and thrombus formation at arterial shear over collagen were attenuated in whole blood treated with ML355 comparable to aspirin. Oral administration of ML355 in mice showed reasonable plasma drug levels by pharmacokinetic assessment. ML355 treatment impaired thrombus growth and vessel occlusion in FeCl 3 -induced mesenteric and laser-induced cremaster arteriole thrombosis models in mice. Importantly, hemostatic plug formation and bleeding after treatment with ML355 was minimal in mice in response to laser ablation on the saphenous vein or in a cremaster microvasculature laser-induced rupture model. Our data strongly support 12-LOX as a key determinant of platelet reactivity in vivo, and inhibition of platelet 12-LOX with ML355 may represent a new class of antiplatelet therapy. © 2017 American Heart Association, Inc.

Test of LOX compatibility for asphalt and concrete runway materials

NASA Technical Reports Server (NTRS)

Moyers, C. V.; Bryan, C. J.; Lockhart, B. J.

1973-01-01

A literature survey and a telephone canvass of producers and users of LOX is reported which yielded one report of an accident resulting from a LOX spill on asphalt, one discussion of hazardous conditions, and an unreferenced mention of an incident. Laboratory tests using standard LOX impact apparatus yielded reactions with both old and new alphalt, but none with concrete. In the final test, using a larger sample of asphalt, the reaction caused extensive damage to equipment. Initial field experiments using 2-meter square asphalt slabs covered with LOX, conducted during rainy weather, achieved no reaction with plummets, and limited reaction with a blasting cap as a reaction initiator. In a final plummet-initiated test on a dry slab, a violent reaction, which appeared to have propagated over the entire slab surface, destroyed the plummet fixture and threw fragments as far as 48 meters.

Methodology for the assessment of oxygen as an energy carrier

NASA Astrophysics Data System (ADS)

Yang, Ming Wei

Due to the energy intensity of the oxygen generating process, the electric power grid would benefit if the oxygen generating process was consumed electric power only during low demand periods. Thus, the question to be addressed in this study is whether oxygen production and/or usage can be modified to achieve energy storage and/or transmission objectives at lower cost. The specific benefit to grid would be a leveling, over time, of the demand profile and thus would require less installation capacity. In order to track the availability of electricity, a compressed air storage unit is installed between the cryogenic distillation section and the main air compressor of air separation unit. A profit maximizing scheme for sizing storage inventory and related equipments is developed. The optimum scheme is capable of market responsiveness. Profits of steel maker, oxy-combustion, and IGCC plants with storage facilities can be higher than those plants without storage facilities, especially, at high-price market. Price tracking feature of air storage integration will certainly increase profit margins of the plants. The integration may push oxy-combustion and integrated gasification combined cycle process into economic viability. Since oxygen is used in consumer sites, it may generate at remote locations and transport to the place needed. Energy losses and costs analysis of oxygen transportation is conducted for various applications. Energy consumptions of large capacity and long distance GOX and LOX pipelines are lower than small capacity pipelines. However, transportation losses and costs of GOX and LOX pipelines are still higher than electricity transmission.

Wavelength-Agile Optical Sensor for Exhaust Plume and Cryogenic Fluid Interrogation

NASA Technical Reports Server (NTRS)

Sanders, Scott T.; Chiaverini, Martin J.; Gramer, Daniel J.

2004-01-01

Two optical sensors developed in UW-Madison labs were evaluated for their potential to characterize rocket engine exhaust plumes and liquid oxygen (LOX) fluid properties. The plume sensor is based on wavelength-agile absorption spectroscopy A device called a chirped white pulse emitter (CWPE) is used to generate the wavelength agile light, scanning, for example, 1340 - 1560 nm every microsecond. Properties of the gases in the rocket plume (for example temperature and water mole fraction) can be monitored using these wavelength scans. We have performed preliminary tests in static gas cells, a laboratory GOX/GH2 thrust chamber, and a solid-fuel hybrid thrust chamber, and these initial tests demonstrate the potential of the CWPE for monitoring rocket plumes. The LOX sensor uses an alternative to wavelength agile sensing: two independent, fixed-wavelength lasers are combined into a single fiber. One laser is absorbed by LOX and the other not: by monitoring the differential transmission the LOX concentration in cryogenic feed lines can be inferred. The sensor was successful in interrogating static LOX pools in laboratory tests. Even in ice- and bubble-laden cryogenic fluids, LOX concentrations were measured to better than 1% with a 3 microsec time constant.

Lipoxygenase-mediated pro-radical effect of melatonin via stimulation of arachidonic acid metabolism

DOE Office of Scientific and Technical Information (OSTI.GOV)

Radogna, F.; Sestili, P.; Martinelli, C.

We have shown that melatonin immediately and transiently stimulates intracellular free radical production on a set of leukocytes, possibly as a consequence of calmodulin binding. We show here that melatonin-induced ROS are produced by lipoxygenase (LOX), since they are prevented by a set of LOX inhibitors, and are accompanied by increase of the 5-LOX product 5-HETE. LOX activation is accompanied by strong liberation of AA; inhibition of Ca{sup 2+}-independent, but not Ca{sup 2+}-dependent, phospholipase A2 (PLA2), prevents both melatonin-induced arachidonic acid and ROS production, whereas LOX inhibition only prevents ROS, indicating that PLA2 is upstream with respect to LOX, asmore » occurs in many signaling pathways. Chlorpromazine, an inhibitor of melatonin-calmodulin interaction, inhibits both ROS and arachidonic acid production, thus possibly placing calmodulin at the origin of a melatonin-induced pro-radical pathway. Interestingly, it is known that Ca{sup 2+}-independent PLA2 binds to calmodulin: our results are compatible with PLA2 being liberated by melatonin from a steady-state calmodulin sequestration, thus initiating an arachidonate signal transduction. These results delineate a novel molecular pathway through which melatonin may participate to the inflammatory response.« less

Thermal inactivation of a recombinant lipoxygenase from Pseudomonas aeruginosa BBE in the absence and presence of additives.

PubMed

Xu, Zhi; Liu, Song; Lu, Xinyao; Rao, Shengqi; Kang, Zhen; Li, Jianghua; Wang, Miao; Chen, Jian

2014-07-01

Bacterial lipoxygenase (EC 1.13.11.12, LOX) is an important enzyme used as a brightener and strengthening agent during breadmaking. In this study, thermal inactivation of a recombinant LOX of Pseudomonas aeruginosa BBE was characterized by kinetic and thermodynamic analysis in the absence and presence of additives. As the heating temperature increased from 25 to 55 °C, the thermal inactivation rate (k) values for LOX without the additives ranged from 0.0407 to 0.2627 min(-1), while the half-life (t1/2) values were between 17.08 and 3.25 min. The activation energy (ΔE) values were increased with rise in heating temperatures from 13.26 to 108.9 kJ mol(-1) . Separate tests at 45 °C in the presence of additives (polyols, sugars and ions) at specific concentrations showed that xylitol (1 mol L(-1)) was the most effective stabilizer for recombinant LOX and increased the t1/2 value by 297%. Recombinant LOX was sensitive to heat treatment, and addition of polyols, sugars and ions could enhance its thermal stability. Our findings may provide useful information for stabilizing emerging bacterial LOXs. © 2013 Society of Chemical Industry.

Coil-On-Plug Ignition for Oxygen/Methane Liquid Rocket Engines in Thermal-Vacuum Environments

NASA Technical Reports Server (NTRS)

Melcher, John C.; Atwell, Matthew J.; Morehead, Robert L.; Hurlbert, Eric A.; Bugarin, Luz; Chaidez, Mariana

2017-01-01

A coil-on-plug ignition system has been developed and tested for Liquid Oxygen (LOX)/liquid methane (LCH4) rocket engines operating in thermal vacuum conditions. The igniters were developed and tested as part of the Integrated Cryogenic Propulsion Test Article (ICPTA), previously tested as part of the Project Morpheus test vehicle. The ICPTA uses an integrated, pressure-fed, cryogenic LOX/LCH4 propulsion system including a reaction control system (RCS) and a main engine. The ICPTA was tested at NASA Glenn Research Center's Plum Brook Station in the Spacecraft Propulsion Research Facility (B-2) under vacuum and thermal vacuum conditions. A coil-on-plug ignition system has been developed to successfully demonstrate ignition reliability at these conditions while preventing corona discharge issues. The ICPTA uses spark plug ignition for both the main engine igniter and the RCS. The coil-on-plug configuration eliminates the conventional high-voltage spark plug cable by combining the coil and the spark plug into a single component. Prior to ICPTA testing at Plum Brook, component-level reaction control engine (RCE) and main engine igniter testing was conducted at NASA Johnson Space Center (JSC), which demonstrated successful hot-fire ignition using the coil-on-plug from sea-level ambient conditions down to 10(exp -2) torr. Integrated vehicle hot-fire testing at JSC demonstrated electrical and command/data system performance. Lastly, hot-fire testing at Plum Brook demonstrated successful ignitions at simulated altitude conditions at 30 torr and cold thermal-vacuum conditions at 6 torr. The test campaign successfully proved that coil-on-plug technology will enable integrated LOX/LCH4 propulsion systems in future spacecraft.

Suppression of allene oxide synthase 3 in potato increases degree of arbuscular mycorrhizal fungal colonization.

PubMed

Morcillo, Rafael Jorge León; Navarrete, María Isabel Tamayo; Bote, Juan Antonio Ocampo; Monguio, Salomé Prat; García-Garrido, José Manuel

2016-01-15

Arbuscular mycorrhizal (AM) is a mutually beneficial interaction among higher plants and soil fungi of the phylum Glomeromycota. Numerous studies have pointed that jasmonic acid plays an important role in the development of the intraradical fungus. This compound belongs to a group of biologically active compounds known as oxylipins which are derived from the oxidative metabolism of polyunsaturated fatty acids. Studies of the regulatory role played by oxylipins in AM colonization have generally focused on jasmonates, while few studies exist on the 9-LOX pathway of oxylipins during AM formation. Here, the cDNA of Allene oxide synthase 3 (AOS3), a key enzyme in the 9-LOX pathway, was used in the RNA interference (RNAi) system to transform potato plants in order to suppress its expression. Results show increases in AOS3 gene expression and 9-LOX products in roots of wild type potato mycorrhizal plants. The suppression of AOS3 gene expression increases the percentage of root with mycorrhizal colonization at early stages of AM formation. AOS3 RNA interference lead to an induction of LOXA and 13-LOX genes, a reduction in AOS3 derived 9-LOX oxylipin compounds and an increase in jasmonic acid content, suggesting compensation between 9 and 13-LOX pathways. The results in a whole support the hypothesis of a regulatory role for the 9-LOX oxylipin pathway during mycorrhization. Copyright © 2015 Elsevier GmbH. All rights reserved.

Liquid oxygen LOX compatibility evaluations of aluminum lithium (Al-Li) alloys: Investigation of the Alcoa 2090 and MMC weldalite 049 alloys

NASA Technical Reports Server (NTRS)

Diwan, Ravinder M.

1989-01-01

The behavior of liquid oxygen (LOX) compatibility of aluminum lithium (Al-Li) alloys is investigated. Alloy systems of Alcoa 2090, vintages 1 to 3, and of Martin Marietta Corporation (MMC) Weldalite 049 were evaluated for their behavior related to the LOX compatibility employing liquid oxygen impact test conditions under ambient pressures and up to 1000 psi. The developments of these aluminum lithium alloys are of critical and significant interest because of their lower densities and higher specific strengths and improved mechanical properties at cryogenic temperatures. Of the different LOX impact tests carried out at the Marshall Space Flight Center (MSFC), it is seen that in certain test conditions at higher pressures, not all Al-Li alloys are LOX compatible. In case of any reactivity, it appears that lithium makes the material more sensitive at grain boundaries due to microstructural inhomogeneities and associated precipitate free zones (PFZ). The objectives were to identify and rationalize the microstructural mechanisms that could be relaxed to LOX compatibility behavior of the alloy system in consideration. The LOX compatibility behavior of Al-Li 2090 and Weldalite 049 is analyzed in detail using microstructural characterization techniques with light optical metallography, scanning electron microscopy (SEM), electron microprobe analysis, and surface studies using secondary ion mass spectrometry (SIMS), electron spectroscopy in chemical analysis (ESCA) and Auger electron spectroscopy (AES). Differences in the behavior of these aluminum lithium alloys are assessed and related to their chemistry, heat treatment conditions, and microstructural effects.

Controlled Cre/loxP Site-Specific Recombination in the Developing Brain in Medaka Fish, Oryzias latipes

PubMed Central

Okuyama, Teruhiro; Isoe, Yasuko; Hoki, Masahito; Suehiro, Yuji; Yamagishi, Genki; Naruse, Kiyoshi; Kinoshita, Masato; Kamei, Yasuhiro; Shimizu, Atushi; Kubo, Takeo; Takeuchi, Hideaki

2013-01-01

Background Genetic mosaic techniques have been used to visualize and/or genetically modify a neuronal subpopulation within complex neural circuits in various animals. Neural populations available for mosaic analysis, however, are limited in the vertebrate brain. Methodology/Principal Findings To establish methodology to genetically manipulate neural circuits in medaka, we first created two transgenic (Tg) medaka lines, Tg (HSP:Cre) and Tg (HuC:loxP-DsRed-loxP-GFP). We confirmed medaka HuC promoter-derived expression of the reporter gene in juvenile medaka whole brain, and in neuronal precursor cells in the adult brain. We then demonstrated that stochastic recombination can be induced by micro-injection of Cre mRNA into Tg (HuC:loxP-DsRed-loxP-GFP) embryos at the 1-cell stage, which allowed us to visualize some subpopulations of GFP-positive cells in compartmentalized regions of the telencephalon in the adult medaka brain. This finding suggested that the distribution of clonally-related cells derived from single or a few progenitor cells was restricted to a compartmentalized region. Heat treatment of Tg(HSP:Cre x HuC:loxP-DsRed-loxP-GFP) embryos (0–1 day post fertilization [dpf]) in a thermalcycler (39°C) led to Cre/loxP recombination in the whole brain. The recombination efficiency was notably low when using 2–3 dpf embyos compared with 0–1 dpf embryos, indicating the possibility of stage-dependent sensitivity of heat-inducible recombination. Finally, using an infrared laser-evoked gene operator (IR-LEGO) system, heat shock induced in a micro area in the developing brains led to visualization of clonally-related cells in both juvenile and adult medaka fish. Conclusions/Significance We established a noninvasive method to control Cre/loxP site-specific recombination in the developing nervous system in medaka fish. This method will broaden the neural population available for mosaic analyses and allow for lineage tracing of the vertebrate nervous system in both juvenile and adult stages. PMID:23825546

Small High-Speed Self-Acting Shaft Seals for Liquid Rocket Engines

NASA Technical Reports Server (NTRS)

Burcham, R. E.; Boynton, J. L.

1977-01-01

Design analysis, fabrication, and experimental evaluation were performed on three self-acting facetype LOX seal designs and one circumferential-type helium deal design. The LOX seals featured Rayleigh step lift pad and spiral groove geometry for lift augmentation. Machined metal bellows and piston ring secondary seal designs were tested. The helium purge seal featured floating rings with Rayleigh step lift pads. The Rayleigh step pad piston ring and the spiral groove LOX seals were successfully tested for approximately 10 hours in liquid oxygen. The helium seal was successfully tested for 24 hours. The shrouded Rayleigh step hydrodynamic lift pad LOX seal is feasible for advanced, small, high-speed oxygen turbopumps.

The 5-lipoxygenase inhibitor tepoxalin induces oxidative damage and altered PTEN status prior to apoptosis in canine osteosarcoma cell lines.

PubMed

Loftus, J P; Cavatorta, D; Bushey, J J; Levine, C B; Sevier, C S; Wakshlag, J J

2016-06-01

The 5-lipoxygenase (5-LOX) inhibitor tepoxalin has been shown to slow canine osteosarcoma (OSA) tumour xenografts growth, yet the mechanisms are poorly elucidated. Further examination of tepoxalin in canine OSA cell lines shows that tepoxalin treated cells undergo apoptosis through caspase-3 activation and annexin staining. Interestingly, apoptosis is superseded by an increase in reactive oxygen species (ROS), as measured by activation of dihydrorhodamine 123 and mitosox. This increase in ROS appears to be related to the 5-LOX inhibitor regardless of cellular 5-LOX status, and was not observed after treatment with the tepoxalin metabolite RWJ20142. Additionally, 5-LOX inhibition by tepoxalin appears to increase phosphatase and tensin (PTEN) homolog activity by preventing its alkylation or oxidation. PTEN modification or inhibition allows phosphoinositide-3 (PI3) kinase activity thereby heightening activation of protein kinase B (AKT) phosphorylation. Our data suggest that off target oxidation and LOX inhibition play roles in the apoptotic response. © 2014 John Wiley & Sons Ltd.

Design, synthesis, and biological evaluation of prenylated chalcones as 5-LOX inhibitors.

PubMed

Reddy, Nimmanapalli P; Aparoy, Polamarasetty; Reddy, T Chandra Mohan; Achari, Chandrani; Sridhar, P Ramu; Reddanna, Pallu

2010-08-15

Ten novel mono- and di-O-prenylated chalcone derivatives were designed on the basis of a homology derived molecular model of 5-lipoxygenase (5-LOX). The compounds were docked into 5-LOX active site and the binding characteristics were quantified using LUDI. To verify our theoretical assumption, the molecules were synthesized and tested for their 5-LOX inhibitory activities. The synthesis was carried out by Claisen-Schmidt condensation reaction of mono- and di-O-prenylated acetophenones with appropriate aldehydes. 5-LOX in vitro inhibition assay showed higher potency of di-O-prenylated chalcones than their mono-O-prenylated chalcone analogs. Compound 5e exhibited good inhibition with an IC(50) at 4 microM. The overall trend for the binding energies calculated and LUDI score was in good qualitative agreement with the experimental data. Further, the compound 5e showed potent anti-proliferative effects (GI(50) at 9 microM) on breast cancer cell line, MCF-7. Copyright 2010 Elsevier Ltd. All rights reserved.

BAC Modification through Serial or Simultaneous Use of CRE/Lox Technology

PubMed Central

Parrish, Mark; Unruh, Jay; Krumlauf, Robb

2011-01-01

Bacterial Artificial Chromosomes (BACs) are vital tools in mouse genomic analyses because of their ability to propagate large inserts. The size of these constructs, however, prevents the use of conventional molecular biology techniques for modification and manipulation. Techniques such as recombineering and Cre/Lox methodologies have thus become heavily relied upon for such purposes. In this work, we investigate the applicability of Lox variant sites for serial and/or simultaneous manipulations of BACs. We show that Lox spacer mutants are very specific, and inverted repeat variants reduce Lox reaction rates through reducing the affinity of Cre for the site, while retaining some functionality. Employing these methods, we produced serial modifications encompassing four independent changes which generated a mouse HoxB BAC with fluorescent reporter proteins inserted into four adjacent Hox genes. We also generated specific, simultaneous deletions using combinations of spacer variants and inverted repeat variants. These techniques will facilitate BAC manipulations and open a new repertoire of methods for BAC and genome manipulation. PMID:21197414

Insights using the molecular model of Lipoxygenase from Finger millet (Eleusine coracana (L.)).

PubMed

Tiwari, Apoorv; Avashthi, Himanshu; Jha, Richa; Srivastava, Ambuj; Kumar Garg, Vijay; Wasudev Ramteke, Pramod; Kumar, Anil

2016-01-01

Lipoxygenase-1 (LOX-1) protein provides defense against pests and pathogens and its presence have been positively correlated with plant resistance against pathogens. Linoleate is a known substrate of lipoxygenase and it induces necrosis leading to the accumulation of isoflavonoid phytoalexins in plant leaves. Therefore, it is of interest to study the structural features of LOX-1 from Finger millet. However, the structure ofLOX-1 from Finger millet is not yet known. A homology model of LOX-1 from Finger millet is described. Domain architecture study suggested the presence of two domains namely PLAT (Phospho Lipid Acyl Transferase) and lipoxygenase. Molecular docking models of linoleate with lipoxygenase from finger millet, rice and sorghum are reported. The features of docked models showed that finger millet have higher pathogen resistance in comparison to other cereal crops. This data is useful for the molecular cloning of fulllength LOX-1 gene for validating its role in improving plant defense against pathogen infection and for various other biological processes.

NSAID loxoprofen inhibits high threshold or wide dynamic range neuronal responses in the rat at different time-courses.

PubMed

Tsuruoka, Masayoshi; Maeda, Masako; Hayashi, Bunsho; Liu, Limin; Inoue, Tomio

2008-01-01

The onset of the antinociceptive effect with loxoprofen sodium (LOX), a non-steroidal anti-inflammatory drug, was examined electrophysiologically during carrageenan-induced hindpaw inflammation in the rat. Extracellular recordings were made from either wide dynamic range (WDR) or high threshold (HT) neurons in the dorsal horn. Recordings from the same neuron were continued for at least 3 h after the injection of carrageenan. Three hours after the induction of inflammation, either a fresh solution of LOX (1 mg/kg) or distilled water was directly administered into the stomach through PE 50 tubing. LOX significantly reduced inflammation-increased background activity and noxious heat-evoked responses in both HT and WDR neurons, whereas distilled water did not produce any change. Asignificant difference in the onset of the inhibitory effect of LOX was observed between HT and WDR neurons. The results show that WDR neurons precede HT neurons regarding inhibition of nociceptive processing in the dorsal horn after administration of LOX.

NESC Review of the 8-Foot High Temperature Tunnel (HTT) Oxygen Storage Pressure Vessel Inspection Requirements

NASA Technical Reports Server (NTRS)

Gilbert, Michael; Raju, Ivatury; Piascik, Robert; Cameron, Kenneth; Kirsch, Michael; Hoffman, Eric; Murthy, Pappu; Hopson, George; Greulich, Owen; Frazier, Wayne

2009-01-01

The 8-Foot HTT (refer to Figure 4.0-1) is used to conduct tests of air-breathing hypersonic propulsion systems at Mach numbers 4, 5, and 7. Methane, Air, and LOX are mixed and burned in a combustor to produce test gas stream containing 21 percent by volume oxygen. The NESC was requested by the NASA LaRC Executive Safety Council to review the rationale for a proposed change to the recertification requirements, specifically the internal inspection requirements, of the 8-Foot HTT LOX Run Tank and LOX Storage Tank. The Run Tank is an 8,000 gallon cryogenic tank used to provide LOX to the tunnel during operations, and is pressured during the tunnel run to 2,250 pounds per square inch gage (psig). The Storage Tank is a 25,000 gallon cryogenic tank used to store LOX at slightly above atmospheric pressure as a external shell, with space between the shells maintained under vacuum conditions.

Oxidant stress in mitochondrial DNA damage, autophagy and inflammation in atherosclerosis

PubMed Central

Ding, Zufeng; Liu, Shijie; Wang, Xianwei; Khaidakov, Magomed; Dai, Yao; Mehta, Jawahar L.

2013-01-01

Our studies in HUVECs show that ox-LDL induced autophagy and damaged mtDNA leading to TLR9 expression. LOX-1 antibody or the ROS inhibitor apocynin attenuated ox-LDL-mediated autophagy, mtDNA damage and TLR9 expression, suggesting that these events are LOX-1 and ROS-dependent phenomena. Experiments using siRNA to DNase II indicated that DNase II digests mtDNA to protect the tissue from inflammation. Next, we studied and found intense autophagy, TLR9 expression and inflammatory signals (CD45 and CD68) in the aortas of LDLR knockout mice fed high cholesterol diet. Deletion of LOX-1 (LDLR/LOX-1 double knockout mice) attenuated autophagy, TLR9 expression as well as CD45 and CD68. Damaged mtDNA signal was also very high in LDLR knockout mice aortas, and this signal was attenuated by LOX-1 deletion. Thus, it appears that oxidative stress-mediated damaged mtDNA that escapes autophagy induces a potent inflammatory response in atherosclerosis. PMID:23326634

Lysyl Oxidase Is Essential for Normal Development and Function of the Respiratory System and for the Integrity of Elastic and Collagen Fibers in Various Tissues

PubMed Central

Mäki, Joni M.; Sormunen, Raija; Lippo, Sari; Kaarteenaho-Wiik, Riitta; Soininen, Raija; Myllyharju, Johanna

2005-01-01

Lysyl oxidases, a family comprising LOX and four LOX-like enzymes, catalyze crosslinking of elastin and collagens. Mouse Lox was recently shown to be crucial for development of the cardiovascular system because null mice died perinatally of aortic aneurysms and cardiovascular dysfunction. We show here that Lox is also essential for development of the respiratory system and the integrity of elastic and collagen fibers in the lungs and skin. The lungs of E18.5 Lox−/− embryos showed impaired development of the distal and proximal airways. Elastic fibers in E18.5 Lox−/− lungs were markedly less intensely stained and more disperse than in the wild type, especially in the mesenchyme surrounding the distal airways, bronchioles, bronchi, and trachea, and were fragmented in pulmonary arterial walls. The organization of individual collagen fibers into tight bundles was likewise abnormal. Similar elastic and collagen fiber abnormalities were seen in the skin. Lysyl oxidase activity in cultured Lox−/− skin fibroblasts and aortic smooth muscle cells was reduced by ∼80%, indicating that Lox is the main isoenzyme in these cells. LOX abnormalities may thus be critical for the pathogenesis of several common diseases, including pulmonary, skin, and cardiovascular disorders. PMID:16192629

Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotl.

PubMed

Stamm, Anne; Strauß, Sarah; Vogt, Peter; Scheper, Thomas; Pepelanova, Iliyana

2018-04-07

AmbLOXe is a lipoxygenase, which is up-regulated during limb-redevelopment in the Mexican axolotl, Ambystoma mexicanum, an animal with remarkable regeneration capacity. Previous studies have shown that mammalian cells transformed with the gene of this epidermal lipoxygenase display faster migration and wound closure rate during in vitro wound healing experiments. In this study, the gene of AmbLOXe was codon-optimized for expression in Escherichia coli and was produced in the insoluble fraction as protein aggregates. These inclusion bodies or nanopills were shown to be reservoirs containing functional protein during in vitro wound healing assays. For this purpose, functional inclusion bodies were used to coat cell culture surfaces prior cell seeding or were added directly to the medium after cells reached confluence. In both scenarios, AmbLOXe inclusion bodies led to faster migration rate and wound closure, in comparison to controls containing either no AmbLOXe or GFP inclusion bodies. Our results demonstrate that AmbLOXe inclusion bodies are functional and may serve as stable reservoirs of this enzyme. Nevertheless, further studies with soluble enzyme are also necessary in order to start elucidating the exact molecular substrates of AmbLOXe and the biochemical pathways involved in the wound healing effect.

XBP1-LOX Axis is critical in ER stress-induced growth of lung adenocarcinoma in 3D culture.

PubMed

Yang, Yi; Cheng, Bai-Jun; Jian, Hong; Chen, Zhi-Wei; Zhao, Yi; Yu, Yong-Feng; Li, Zi-Ming; Liao, Mei-Lin; Lu, Shun

2017-01-01

Rapid growth of tumor cells needs to consume large amounts of oxygen and glucose, due to lack of blood supply within the tumor, cells live in an environment that lack of oxygen and nutrients. This environment results in endoplasmic reticulum (ER) stress and activates the UPR (unfolded protein response). More and more evidence suggests UPR provides a growth signal pathway required for tumor growth. In the present study, we investigated the relationship between XBP1, one transcription factor in UPR, and the expression of LOX. We found that ER stress induces high expression of XBP1, one transcription factor in UPR, in both 2D culture and 3D culture; but only promotes growth of lung adenocarcinoma cells in in vitro 3D culture other than 2D culture. In 3D culture, we further showed that knockdown XBP1 expression can block Tm/Tg-induced cell growth. LOX genes may be key downstream effector of XBP1. Knockdown LOX expression can partially block XBP1-induced cell growth. Then we showed XBP1 suppressed by RNA interference (RNAi) can reduce the expression of LOX. For the first time, it is being shown that XBP1 can regulate the expression of LOX to promote cell growth.

Pharmacological profile and efficiency in vivo of diflapolin, the first dual inhibitor of 5-lipoxygenase-activating protein and soluble epoxide hydrolase.

PubMed

Garscha, Ulrike; Romp, Erik; Pace, Simona; Rossi, Antonietta; Temml, Veronika; Schuster, Daniela; König, Stefanie; Gerstmeier, Jana; Liening, Stefanie; Werner, Markus; Atze, Heiner; Wittmann, Sandra; Weinigel, Christina; Rummler, Silke; Scriba, Gerhard K; Sautebin, Lidia; Werz, Oliver

2017-08-24

Arachidonic acid (AA) is metabolized to diverse bioactive lipid mediators. Whereas the 5-lipoxygenase-activating protein (FLAP) facilitates AA conversion by 5-lipoxygenase (5-LOX) to pro-inflammatory leukotrienes (LTs), the soluble epoxide hydrolase (sEH) degrades anti-inflammatory epoxyeicosatrienoic acids (EETs). Accordingly, dual FLAP/sEH inhibition might be advantageous drugs for intervention of inflammation. We present the in vivo pharmacological profile and efficiency of N-[4-(benzothiazol-2-ylmethoxy)-2-methylphenyl]-N'-(3,4-dichlorophenyl)urea (diflapolin) that dually targets FLAP and sEH. Diflapolin inhibited 5-LOX product formation in intact human monocytes and neutrophils with IC 50  = 30 and 170 nM, respectively, and suppressed the activity of isolated sEH (IC 50  = 20 nM). Characteristic for FLAP inhibitors, diflapolin (I) failed to inhibit isolated 5-LOX, (II) blocked 5-LOX product formation in HEK cells only when 5-LOX/FLAP was co-expressed, (III) lost potency in intact cells when exogenous AA was supplied, and (IV) prevented 5-LOX/FLAP complex assembly in leukocytes. Diflapolin showed target specificity, as other enzymes related to AA metabolism (i.e., COX1/2, 12/15-LOX, LTA 4 H, LTC 4 S, mPGES 1 , and cPLA 2 ) were not inhibited. In the zymosan-induced mouse peritonitis model, diflapolin impaired vascular permeability, inhibited cysteinyl-LTs and LTB 4 formation, and suppressed neutrophil infiltration. Diflapolin is a highly active dual FLAP/sEH inhibitor in vitro and in vivo with target specificity to treat inflammation-related diseases.

Vascular lysyl oxidase over-expression alters extracellular matrix structure and induces oxidative stress.

PubMed

Varona, Saray; García-Redondo, Ana B; Martínez-González, Jose; Salaices, Mercedes; Briones, Ana M; Rodríguez, Cristina

Lysyl oxidase (LOX) participates in the assembly of collagen and elastin fibres. The impact of vascular LOX over-expression on extracellular matrix (ECM) structure and its contribution to oxidative stress has been analysed. Studies were conducted on mice over-expressing LOX (Tg), specifically in smooth muscle cells (VSMC). Gene expression was assessed by real-time PCR analysis. Sirius Red staining, H 2 O 2 production and NADPH oxidase activity were analysed in different vascular beds. The size and number of fenestra of the internal elastic lamina were determined by confocal microscopy. LOX activity was up-regulated in VSMC of transgenic mice compared with cells from control animals. At the same time, transgenic cells deposited more organised elastin fibres and their supernatants induced a stronger collagen assembly in in vitro assays. Vascular collagen cross-linking was also higher in Tg mice, which showed a decrease in the size of fenestrae and an enhanced expression of Fibulin-5. Interestingly, higher H 2 O 2 production and NADPH oxidase activity was detected in the vascular wall from transgenic mice. The H 2 O 2 scavenger catalase attenuated the stronger deposition of mature elastin fibres induced by LOX transgenesis. LOX over-expression in VSMC was associated with a change in the structure of collagen and elastin fibres. LOX could constitute a novel source of oxidative stress that might participate in elastin changes and contribute to vascular remodelling. Copyright © 2017 Sociedad Española de Arteriosclerosis. Publicado por Elsevier España, S.L.U. All rights reserved.

Function of the Pyruvate Oxidase-Lactate Oxidase Cascade in Interspecies Competition between Streptococcus oligofermentans and Streptococcus mutans

PubMed Central

Liu, Lei

2012-01-01

Complex interspecies interactions occur constantly between oral commensals and the opportunistic pathogen Streptococcus mutans in dental plaque. Previously, we showed that oral commensal Streptococcus oligofermentans possesses multiple enzymes for H2O2 production, especially lactate oxidase (Lox), allowing it to out-compete S. mutans. In this study, through extensive biochemical and genetic studies, we identified a pyruvate oxidase (pox) gene in S. oligofermentans. A pox deletion mutant completely lost Pox activity, while ectopically expressed pox restored activity. Pox was determined to produce most of the H2O2 in the earlier growth phase and log phase, while Lox mainly contributed to H2O2 production in stationary phase. Both pox and lox were expressed throughout the growth phase, while expression of the lox gene increased by about 2.5-fold when cells entered stationary phase. Since lactate accumulation occurred to a large degree in stationary phase, the differential Pox- and Lox-generated H2O2 can be attributed to differential gene expression and substrate availability. Interestingly, inactivation of pox causes a dramatic reduction in H2O2 production from lactate, suggesting a synergistic action of the two oxidases in converting lactate into H2O2. In an in vitro two-species biofilm experiment, the pox mutant of S. oligofermentans failed to inhibit S. mutans even though lox was active. In summary, S. oligofermentans develops a Pox-Lox synergy strategy to maximize its H2O2 formation so as to win the interspecies competition. PMID:22287002

Expression of ESR1 in Glutamatergic and GABAergic Neurons Is Essential for Normal Puberty Onset, Estrogen Feedback, and Fertility in Female Mice.

PubMed

Cheong, Rachel Y; Czieselsky, Katja; Porteous, Robert; Herbison, Allan E

2015-10-28

Circulating estradiol exerts a profound influence on the activity of the gonadotropin-releasing hormone (GnRH) neuronal network controlling fertility. Using genetic strategies enabling neuron-specific deletion of estrogen receptor α (Esr1), we examine here whether estradiol-modulated GABA and glutamate transmission are critical for the functioning of the GnRH neuron network in the female mouse. Using Vgat- and Vglut2-ires-Cre knock-in mice and ESR1 immunohistochemistry, we demonstrate that subpopulations of GABA and glutamate neurons throughout the limbic forebrain express ESR1, with ESR1-GABAergic neurons being more widespread and numerous than ESR1-glutamatergic neurons. We crossed Vgat- and Vglut2-ires-Cre mice with an Esr1(lox/lox) line to generate animals with GABA-neuron-specific or glutamate-neuron-specific deletion of Esr1. Vgat-ires-Cre;Esr1(lox/lox) mice were infertile, with abnormal estrous cycles, and exhibited a complete failure of the estrogen positive feedback mechanism responsible for the preovulatory GnRH surge. However, puberty onset and estrogen negative feedback were normal. Vglut2-ires-Cre;Esr1(lox/lox) mice were also infertile but displayed a wider range of deficits, including advanced puberty onset, abnormal negative feedback, and abolished positive feedback. Whereas <25% of preoptic kisspeptin neurons expressed Cre in Vgat- and Vglut2-ires-Cre lines, ∼70% of arcuate kisspeptin neurons were targeted in Vglut2-ires-Cre;Esr1(lox/lox) mice, possibly contributing to their advanced puberty phenotype. These observations show that, unexpectedly, ESR1-GABA neurons are only essential for the positive feedback mechanism. In contrast, we reveal the key importance of ESR1 in glutamatergic neurons for multiple estrogen feedback loops within the GnRH neuronal network required for fertility in the female mouse. Copyright © 2015 the authors 0270-6474/15/3514533-11$15.00/0.

Arsenic augments the uptake of oxidized LDL by upregulating the expression of lectin-like oxidized LDL receptor in mouse aortic endothelial cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hossain, Ekhtear; Ota, Akinobu, E-mail: aota@aichi-med-u.ac.jp; Karnan, Sivasundaram

Although chronic arsenic exposure is a well-known risk factor for cardiovascular diseases, including atherosclerosis, the molecular mechanism underlying arsenic-induced atherosclerosis remains obscure. Therefore, this study aimed to elucidate this molecular mechanism. We examined changes in the mRNA level of the lectin-like oxidized LDL (oxLDL) receptor (LOX-1) in a mouse aortic endothelial cell line, END-D, after sodium arsenite (SA) treatment. SA treatment significantly upregulated LOX-1 mRNA expression; this finding was also verified at the protein expression level. Flow cytometry and fluorescence microscopy analyses showed that the cellular uptake of fluorescence (Dil)-labeled oxLDL was significantly augmented with SA treatment. In addition, anmore » anti-LOX-1 antibody completely abrogated the augmented uptake of Dil-oxLDL. We observed that SA increased the levels of the phosphorylated forms of nuclear factor of kappa light polypeptide gene enhancer in B cells (NF-κB)/p65. SA-induced upregulation of LOX-1 protein expression was clearly prevented by treatment with an antioxidant, N-acetylcysteine (NAC), or an NF-κB inhibitor, caffeic acid phenethylester (CAPE). Furthermore, SA-augmented uptake of Dil-oxLDL was also prevented by treatment with NAC or CAPE. Taken together, our results indicate that arsenic upregulates LOX-1 expression through the reactive oxygen species-mediated NF-κB signaling pathway, followed by augmented cellular oxLDL uptake, thus highlighting a critical role of the aberrant LOX-1 signaling pathway in the pathogenesis of arsenic-induced atherosclerosis. - Highlights: • Sodium arsenite (SA) increases LOX-1 expression in mouse aortic endothelial cells. • SA enhances cellular uptake of oxidized LDL in dose-dependent manner. • SA-induced ROS generation enhances phosphorylation of NF-κB. • SA upregulates LOX-1 expression through ROS-activated NF-κB signaling pathway.« less

[Discover potential inhibitors of 5-LOX and LTA4H from Rhei Radix et Rhizoma, Notopterygii Rhizoma et Radix and Genitana Macrophyllae Radix based on molecular simulation methods].

PubMed

Gu, Yu; Zhang, Xu; Chen, Yan-Kun; Zhao, Bo-Wen; Zhang, Yan-Ling

2017-12-01

5-lipoxygenase (5-LOX) and leukotriene A4 hydrolase (LTA4H), as the major targets of 5-LOX branch in the arachidonic acid (AA) metabolic pathway, play an important role in the treatment of inflammation. Rhei Radix et Rhizoma, Notopterygii Rhizoma et Radix and Genitana Macrophyllae Radix have clear anti-inflammation activities. In this paper, the targets of 5-LOX and LTA4H were used as the research carrier, and Hiphop module in DS4.0 (Discovery studio) was used to construct ingredients database for preliminary screening of three traditional Chinese medicines based on target inhibitor pharmacophore, so as to obtain 5-LOX and LTA4H potential active ingredients. The ingredients obtained in initial pharmacophore screening were further screened by using CDOCKER module, and the screening rules were established based on the score of initial compound and the key amino acids to obtain 12 potential 5-LOX inhibitors and 7 potential LTA4H inhibitors. To be more specific, the potential 5-LOX inhibitors included 6 ingredients in Rhei Radix et Rhizoma, such as procyanidins B2-3,3'-O-double gallate and revandchinone 2; four ingredients in notopterygium, such as dodecanoic acid and so on. On the other hand, potential LTA4H inhibitors included revandchinone 1, revandchinone 4 in Rhei Radix et Rhizoma, tridecanoic acid, tetracosanoic acid and methyl eicosanoate in Notopterygii Rhizoma et Radix, montanic acid methyl ester and N-docosanoyl-O-aminobenzoate in Genitana Macrophyllae Radix and so on. The molecular simulation methods were highly efficient and time-saving to obtain the potential inhibitors of 5-LOX and LTA4H, which could provide assistance for discovering the chemical quality indicators of anti-inflammatory efficacy of three Chinese herbs, and may be helpful to promote the whole-process quality control of three Chinese herbs. Copyright© by the Chinese Pharmaceutical Association.

Ignition Characterization Test Results for the LO2/Ethanol Propellant Combination

NASA Technical Reports Server (NTRS)

Popp, Christopher G.; Robinson, Phillip J.; Veith, Eric M.

2006-01-01

A series of contracts were issued by the Marshall Space Flight Center (MSFC) of the National Aeronautics and Space Administration (NASA) under the auspices of the Exploration Systems Mission Directorate to develop and expand the maturity of candidate technologies considered to be important for future space exploration. One such technology was to determine the viability of incorporating non-toxic propellants for Reaction Control Subsystems (RCS). Contract NAS8-01109 was issued to Aerojet to develop a dual thrust Reaction Control Engine (RCE) that utilized liquid oxygen and ethanol as the propellants. The dual thrust RCE incorporated a primary thrust level of 870 lbf, and a vernier thrust level of 10 - 30 lbf. The preferred RCS approach for the dual thrust RCE was to utilize pressure-fed liquid oxygen (LOX) and ethanol propellants; however, previous dual thrust feasibility testing incorporated GOX/Ethanol igniters as opposed to LOX/Ethanol igniters in the design. GOX/Ethanol was easier to ignite, but this combination had system design implications of providing GOX for the igniters. A LOX/Ethanol igniter was desired; however, extensive LOX/Ethanol ignition data over the anticipated operating range for the dual thrust RCE did not exist. Therefore, Aerojet designed and tested a workhorse LOX igniter to determine LOX/Ethanol ignition characteristics as part of a risk mitigation effort for the dual thrust RCE design. LOX, encompassing potential two-phase flow conditions anticipated being present in real mission applications. A workhorse igniter was designed to accommodate the hll LOX design flowrate, as well as a reduced GOX flowrate. It was reasoned that the initial LOX flow through the igniter would flash to GOX due to the latent heat stored in the hardware, causing a reduced oxygen flowrate because of a choked, or sonic, flow condition through the injection elements. As LOX flow continued, the hardware would chill-in, with the injected oxygen flow transitioning from cold GOX through two-phase flow to subcooled LOX. permitted oxygen state points to be determined in the igniter oxidizer manifold, and gas-side igniter chamber thermocouples provided chamber thermal profile characteristics. The cold flow chamber pressure (P(sub c)) for each test was determined and coupled with the igniter chamber diameter (D(sub c)) to calculate the characteristic quench parameter (P(sub c) x D(sub c)), which was plotted as a function of core mixture ratio, m. Ignition limits were determined over a broad range of valve inlet conditions, and ignition was demonstrated with oxygen inlet conditions that ranged from subcooled 210 R LOX to 486 R GOX. Once ignited at cold GOX conditions, combustion was continuous as the hardware chilled in and the core mixture ratio transitioned from values near 1.0 to over 12.5. Pulsing is required in typical RCS engines; therefore, the workhorse igniter was pulse tested to verify the ability to provide the required ignition for a pulsing RCE. The minimum electrical pulse width (EPW) of the dual thrust RCE was 0.080 seconds.

Expression of collagen and related growth factors in rat tendon and skeletal muscle in response to specific contraction types

PubMed Central

Heinemeier, K M; Olesen, J L; Haddad, F; Langberg, H; Kjaer, M; Baldwin, K M; Schjerling, P

2007-01-01

Acute exercise induces collagen synthesis in both tendon and muscle, indicating an adaptive response in the connective tissue of the muscle–tendon unit. However, the mechanisms of this adaptation, potentially involving collagen-inducing growth factors (such as transforming growth factor-β-1 (TGF-β-1)), as well as enzymes related to collagen processing, are not clear. Furthermore, possible differential effects of specific contraction types on collagen regulation have not been investigated. Female Sprague–Dawley rats were subjected to 4 days of concentric, eccentric or isometric training (n = 7–9 per group) of the medial gastrocnemius, by stimulation of the sciatic nerve. RNA was extracted from medial gastrocnemius and Achilles tendon tissue 24 h after the last training bout, and mRNA levels for collagens I and III, TGF-β-1, connective tissue growth factor (CTGF), lysyl oxidase (LOX), metalloproteinases (MMP-2 and -9) and their inhibitors (TIMP-1 and 2) were measured by Northern blotting and/or real-time PCR. In tendon, expression of TGF-β-1 and collagens I and III (but not CTGF) increased in response to all types of training. Similarly, enzymes/factors involved in collagen processing were induced in tendon, especially LOX (up to 37-fold), which could indicate a loading-induced increase in cross-linking of tendon collagen. In skeletal muscle, a similar regulation of gene expression was observed, but in contrast to the tendon response, the effect of eccentric training was significantly greater than the effect of concentric training on the expression of several transcripts. In conclusion, the study supports an involvement of TGF-β-1 in loading-induced collagen synthesis in the muscle–tendon unit and importantly, it indicates that muscle tissue is more sensitive than tendon to the specific mechanical stimulus. PMID:17540706

Expression of collagen and related growth factors in rat tendon and skeletal muscle in response to specific contraction types.

PubMed

Heinemeier, K M; Olesen, J L; Haddad, F; Langberg, H; Kjaer, M; Baldwin, K M; Schjerling, P

2007-08-01

Acute exercise induces collagen synthesis in both tendon and muscle, indicating an adaptive response in the connective tissue of the muscle-tendon unit. However, the mechanisms of this adaptation, potentially involving collagen-inducing growth factors (such as transforming growth factor-beta-1 (TGF-beta-1)), as well as enzymes related to collagen processing, are not clear. Furthermore, possible differential effects of specific contraction types on collagen regulation have not been investigated. Female Sprague-Dawley rats were subjected to 4 days of concentric, eccentric or isometric training (n = 7-9 per group) of the medial gastrocnemius, by stimulation of the sciatic nerve. RNA was extracted from medial gastrocnemius and Achilles tendon tissue 24 h after the last training bout, and mRNA levels for collagens I and III, TGF-beta-1, connective tissue growth factor (CTGF), lysyl oxidase (LOX), metalloproteinases (MMP-2 and -9) and their inhibitors (TIMP-1 and 2) were measured by Northern blotting and/or real-time PCR. In tendon, expression of TGF-beta-1 and collagens I and III (but not CTGF) increased in response to all types of training. Similarly, enzymes/factors involved in collagen processing were induced in tendon, especially LOX (up to 37-fold), which could indicate a loading-induced increase in cross-linking of tendon collagen. In skeletal muscle, a similar regulation of gene expression was observed, but in contrast to the tendon response, the effect of eccentric training was significantly greater than the effect of concentric training on the expression of several transcripts. In conclusion, the study supports an involvement of TGF-beta-1 in loading-induced collagen synthesis in the muscle-tendon unit and importantly, it indicates that muscle tissue is more sensitive than tendon to the specific mechanical stimulus.

Ignition Characterization Tests of the LOX/Ethanol Propellant Combination

NASA Technical Reports Server (NTRS)

Popp, Christopher G.; Robinson, Philip J.; Veith, Eric M.

2004-01-01

A series of contracts have been issued by the Marshall Space Flight Center (MSFC) of the National Aeronautics and Space Administration (NASA) to explore candidate technologies considered to be important for the Next Generation Launch Technology (NGLT) effort. One aspect of the NGLT effort is to explore the potential of incorporating non-toxic propellants for Reaction Control Subsystems (RCS). Contract NAS8-01109 has been issued to Aerojet to develop a dual thrust Reaction Control Engine (RCE) that utilizes liquid oxygen and ethanol as the propellants. The dual thrust RCE incorporates a primary thrust level of 870 lbf, and a vernier thrust level of 10 - 30 lbf. Aerojet has designed and tested a workhorse LOX igniter to determine LOX/Ethanol ignition characteristics as part of a risk mitigation effort for the dual thrust RCE design. The objective of the ignition testing was to demonstrate successfid ignition from GOX to LOX, encompassing potential two-phase flow conditions. The workhorse igniter was designed to accommodate the full LOX design flowrate, as well as a reduced GOX flowrate. It was reasoned that the initial LOX flow through the igniter would flash to GOX due to the inherent heat stored in the hardware, causing a reduced oxygen flowrate because of a choked, or sonic, flow condition through the injection elements. As LOX flow continued, the inherent heat of the test hardware would be removed and the hardware would chill-in, with the injected oxygen flow transitioning from cold GOX through two-phase flow to subcooled LOX. Pressure and temperature instrumentation permitted oxygen state points to be determined, and gas-side igniter chamber thermocouples provided chamber thermal profile characteristics. The cold flow chamber pressure (P(sub c)) for each test was determined and coupled with the igniter chamber diameter (D(sub c)) to calculate the characteristic quench parameter (P(sub c) x D(sub c)), which was plotted as a function of core mixture ratio, MR(sub c). Ignition limits were determined over a broad range of valve inlet conditions, and ignition was demonstrated with oxygen inlet conditions that ranged from subcooled 173 R LOX to 480 R GQX. Once ignited at cold GOX conditions, combustion was continuous as the hardware chilled in and the core mixture ratio transitioned from values near 1.0 to over 12.5.

Xanthine Oxidase Induces Foam Cell Formation through LOX-1 and NLRP3 Activation.

PubMed

Dai, Yao; Cao, Yongxiang; Zhang, Zhigao; Vallurupalli, Srikanth; Mehta, Jawahar L

2017-02-01

Xanthine oxidase catalyzes the oxidation of xanthine to uric acid. This process generates excessive reactive oxygen species (ROS) that play an important role in atherogenesis. Recent studies show that LRR and PYD domains-containing protein 3 (NLRP3), a component of the inflammasome, may be involved in the formation of foam cells, a hallmark of atherosclerosis. This study was designed to study the role of various scavenger receptors and NLRP3 inflammasome in xanthine oxidase and uric acid-induced foam cell formation. Human vascular smooth muscle cells (VSMCs) and THP-1 macrophages were treated with xanthine oxidase or uric acid. Xanthine oxidase treatment (of both VSMCs and THP-1 cells) resulted in foam cell formation in concert with generation of ROS and expression of cluster of differentiation 36 (CD36) and oxidized low density lipoprotein (lectin-like) receptor 1 (LOX-1), but not of scavenger receptor A (SRA). Uric acid treatment resulted in foam cell formation, ROS generation and expression of CD36, but not of LOX-1 or SRA. Further, treatment of cells with xanthine oxidase, but not uric acid, activated NLRP3 and its downstream pro-inflammatory signals- caspase-1, interleukin (IL)-1β and IL-18. Blockade of LOX-1 or NLRP3 inflammasome with specific siRNAs reduced xanthine oxidase-induced foam cell formation, ROS generation and activation of NLRP3 and downstream signals. Xanthine oxidase induces foam cell formation in large part through activation of LOX-1 - NLRP3 pathway in both VSMCs and THP-1 cells, but uric acid-induced foam cell formation is exclusively through CD36 pathway. Further, LOX-1 activation is upstream of NLRP3 activation. Graphical Abstract Steps in the formation of foam cells in response to xanthine oxidase and uric acid. Xanthine oxidase stimulates LOX-1 expression on the cell membrane of macrophages and vascular smooth muscle cells (VSMCs) and increases generation of ROS, which activate NLRP3 inflammasome and downstream pro-inflammatory mediators such as Caspase-1, IL-1β and IL-18. Xanthine oxidase also induces CD36 expression. Activation of both LOX-1 and CD36 (LOX-1> > CD36) participates in the transformation of macrophages and VSMCs into foam cells. Uric acid formed from xanthine-xanthine oxidase interaction stimulates CD36 expression and triggers foam cell formation independent of NLRP3 activation.

MiR-29b inhibits collagen maturation in hepatic stellate cells through down-regulating the expression of HSP47 and lysyl oxidase

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Yifei; Ghazwani, Mohammed; Li, Jiang

Highlights: • Enhanced HSP47 and LOX expression is associated with decreased miR-29b level in liver fibrosis. • miR-29b down-regulates HSP47 and LOX expression. • The suppression of HSP47 and LOX by miR-29b is mediated by putative sites at their 3′-UTRs. • miR-29b inhibits extracellular LOX activity and collagen maturation. - Abstract: Altered expression of miR-29b is implicated in the pathogenesis and progression of liver fibrosis. We and others previously demonstrated that miR-29b down-regulates the expression of several extracellular-matrix (ECM) genes including Col 1A1, Col 3A1 and Elastin via directly targeting their 3′-UTRs. However, whether or not miR-29b plays a rolemore » in the post-translational regulation of ECM biosynthesis has not been reported. Heat shock protein 47 (HSP47) and lysyl oxidase (LOX) are known to be essential for ECM maturation. In this study we have demonstrated that expression of HSP47 and LOX was significantly up-regulated in culture-activated primary rat hepatic stellate cells (HSCs), TGF-β stimulated LX-2 cells and liver tissue of CCl{sub 4}-treated mice, which was accompanied by a decrease of miR-29b level. In addition, over-expression of miR-29b in LX-2 cells resulted in significant inhibition on HSP47 and LOX expression. Mechanistically, miR-29b inhibited the expression of a reporter gene that contains the respective full-length 3′-UTR from HSP47 and LOX gene, and this inhibitory effect was abolished by the deletion of a putative miR-29b targeting sequence from the 3′-UTRs. Transfection of LX-2 cells with miR-29b led to abnormal collagen structure as shown by electron-microscopy, presumably through down-regulation of the expression of molecules involved in ECM maturation including HSP47 and LOX. These results demonstrated that miR-29b is involved in regulating the post-translational processing of ECM and fibril formation.« less

Human 15-LOX-1 active site mutations alter inhibitor binding and decrease potency.

PubMed

Armstrong, Michelle; van Hoorebeke, Christopher; Horn, Thomas; Deschamps, Joshua; Freedman, J Cody; Kalyanaraman, Chakrapani; Jacobson, Matthew P; Holman, Theodore

2016-11-01

Human 15-lipoxygenase-1 (h15-LOX-1 or h12/15-LOX) reacts with polyunsaturated fatty acids and produces bioactive lipid derivatives that are implicated in many important human diseases. One such disease is stroke, which is the fifth leading cause of death and the first leading cause of disability in America. The discovery of h15-LOX-1 inhibitors could potentially lead to novel therapeutics in the treatment of stroke, however, little is known about the inhibitor/active site interaction. This study utilizes site-directed mutagenesis, guided in part by molecular modeling, to gain a better structural understanding of inhibitor interactions within the active site. We have generated eight mutants (R402L, R404L, F414I, F414W, E356Q, Q547L, L407A, I417A) of h15-LOX-1 to determine whether these active site residues interact with two h15-LOX-1 inhibitors, ML351 and an ML094 derivative, compound 18. IC 50 values and steady-state inhibition kinetics were determined for the eight mutants, with four of the mutants affecting inhibitor potency relative to wild type h15-LOX-1 (F414I, F414W, E356Q and L407A). The data indicate that ML351 and compound 18, bind in a similar manner in the active site to an aromatic pocket close to F414 but have subtle differences in their specific binding modes. This information establishes the binding mode for ML094 and ML351 and will be leveraged to develop next-generation inhibitors. Copyright © 2016 Elsevier Ltd. All rights reserved.

Chronic effects of aerobic exercise on gene expression of LOX-1 receptor in the heart of rats fed with high fat diet

PubMed Central

Riahi, Simin; Mohammadi, Mohammad Taghi; Sobhani, Vahid; Soleimany, Mansureh

2015-01-01

Objective(s): Lectin-like low density lipoprotein receptor (LOX-1) has pivot role in vascular complications, which is upregulated in numerous pathological conditions. Since exercise has beneficial effects in prevention of hyperlipidemic complications, present study examined protective effects of aerobic exercise through reduction of LOX-1 expression in heart during dyslipidemia. Materials and Methods: Four groups of rats were used (N=25): Normal, Normal and exercise, High fat and High fat and exercise. High fat diet (HFD) was made by adding 10% animal oil, 2% cholesterol and 0.5% colic acid to standard rodent chow. Exercise protocol consisted of swimming 1 hr/day, and 5 days/week for 8 weeks. Plasma lipids were evaluated at the end of experiment, 48 hr after final session of exercise. At the end, rats were sacrificed and heart was removed for determination of malondialdehyde (MDA) content, and LOX-1 expression. Results: HFD meaningfully changed lipid profile (>50%), but chronic exercise had no significant effects on lipid profile. LOX-1 expression was significantly increased in heart of rats fed with HFD, while swimming exercise considerably reduced gene expression of LOX-1. MDA content was significantly enhanced in rats fed with HFD (4.37±0.6 nmol/mg, P<0.01) compared to normal group (1.56±0.48 nmol/mg), whereas swimming exercise decreased MDA level of heart in rats fed with HFD (2.28±0.32, P<0.01). Conclusion: Findings indicated that swimming exercise is able to diminish heart expression of LOX-1 receptor concomitant reduction of oxidative stress. Since these parameters are involved in generation of dyslipidemic complications, swimming exercise is a good candidate to reduce these complications. PMID:26557970

PgLOX6 encoding a lipoxygenase contributes to jasmonic acid biosynthesis and ginsenoside production in Panax ginseng

PubMed Central

Rahimi, Shadi; Kim, Yu-Jin; Sukweenadhi, Johan; Zhang, Dabing; Yang, Deok-Chun

2016-01-01

Ginsenosides, the valuable pharmaceutical compounds in Panax ginseng, are triterpene saponins that occur mainly in ginseng plants. It was shown that in vitro treatment with the phytohormone jasmonic acid (JA) is able to increase ginsenoside production in ginseng plants. To understand the molecular link between JA biosynthesis and ginsenoside biosynthesis, we identified a JA biosynthetic 13-lipoxygenase gene (PgLOX6) in P. ginseng that promotes ginsenoside production. The expression of PgLOX6 was high in vascular bundles, which corresponds with expression of ginsenoside biosynthetic genes. Consistent with the role of PgLOX6 in synthesizing JA and promoting ginsenoside synthesis, transgenic plants overexpressing PgLOX6 in Arabidopsis had increased amounts of JA and methyl jasmonate (MJ), increased expression of triterpene biosynthetic genes such as squalene synthase (AtSS1) and squalene epoxidase (AtSE1), and increased squalene content. Moreover, transgenic ginseng roots overexpressing PgLOX6 had around 1.4-fold increased ginsenoside content and upregulation of ginsenoside biosynthesis-related genes including PgSS1, PgSE1, and dammarenediol synthase (PgDDS), which is similar to that of treatment with MJ. However, MJ treatment of transgenic ginseng significantly enhanced JA and MJ, associated with a 2.8-fold increase of ginsenoside content compared with the non-treated, non-transgenic control plant, which was 1.4 times higher than the MJ treatment effect on non-transgenic plants. These results demonstrate that PgLOX6 is responsible for the biosynthesis of JA and promotion of the production of triterpenoid saponin through up-regulating the expression of ginsenoside biosynthetic genes. This work provides insight into the role of JA in biosynthesizing secondary metabolites and provides a molecular tool for increasing ginsenoside production. PMID:27811076

Silencing IL-13Rα2 promotes glioblastoma cell death via endogenous signaling.

PubMed

Hsi, Linda C; Kundu, Suman; Palomo, Juan; Xu, Bo; Ficco, Ryan; Vogelbaum, Michael A; Cathcart, Martha K

2011-07-01

Glioblastoma multiforme (GBM) is one of the most lethal forms of cancer, with a survival rate of only 13% to 27% within 2 years of diagnosis despite optimal medical treatment. We hypothesize that the presence of a unique IL-13Rα2 decoy receptor prevents GBM apoptosis. This receptor has a high affinity for interleukin-13 (IL-13), binds the cytokine, and competitively inhibits the intracellular signaling cascade initiated by IL-13. In cells lacking the IL-13Rα2 decoy receptor, IL-13 initiates the production of 15-lipoxygenase-1 (15-LOX-1), which has been implicated in cellular apoptosis. Our group and others have shown that induction of 15-LOX-1 correlates with tumor cell death in colorectal, pancreatic, and prostate cancer. How 15-LOX-1 induces apoptosis remains unclear. Preliminary evidence in GBM cells implicates an apoptotic process mediated by PPARγ. 15-LOX-1 metabolites can modulate PPARγ and activation of PPARγ can suppress tumor growth. We hypothesize that in GBM, IL-13 can induce 15-LOX-1, which regulates cell apoptosis via signaling through PPARγ and that expression of IL-13Rα2 prevents apoptosis and contributes to tumor growth. Our in vitro and in vivo data support this. Knocking down IL-13Rα2 with short interfering RNA dramatically induces 15-LOX-1 expression, promotes apoptosis, and reduces GBM tumor growth in vivo. These findings identify a mechanism for eliminating the blockade of endogenous IL-13 signaling and for promotion of apoptosis, and characterize a role for 15-LOX-1 in GBM apoptosis. Identifying a mechanistic pathway that can be targeted for pharmacologic intervention will have applied implications to developing novel and effective treatments of GBM. © 2011 American Association for Cancer Research.

Characterization of the maize lipoxygenase gene family in relation to aflatoxin accumulation resistance.

PubMed

Ogunola, Oluwaseun F; Hawkins, Leigh K; Mylroie, Erik; Kolomiets, Michael V; Borrego, Eli; Tang, Juliet D; Williams, W Paul; Warburton, Marilyn L

2017-01-01

Maize (Zea mays L.) is a globally important staple food crop prone to contamination by aflatoxin, a carcinogenic secondary metabolite produced by the fungus Aspergillus flavus. An efficient approach to reduce accumulation of aflatoxin is the development of germplasm resistant to colonization and toxin production by A. flavus. Lipoxygenases (LOXs) are a group of non-heme iron containing dioxygenase enzymes that catalyze oxygenation of polyunsaturated fatty acids (PUFAs). LOX derived oxylipins play critical roles in plant defense against pathogens including A. flavus. The objectives of this study were to summarize sequence diversity and expression patterns for all LOX genes in the maize genome, and map their effect on aflatoxin accumulation via linkage and association mapping. In total, 13 LOX genes were identified, characterized, and mapped. The sequence of one gene, ZmLOX10, is reported from 5 inbred lines. Genes ZmLOX1/2, 5, 8, 9, 10 and 12 (GRMZM2G156861, or V4 numbers ZM00001D042541 and Zm00001D042540, GRMZM2G102760, GRMZM2G104843, GRMZM2G017616, GRMZM2G015419, and GRMZM2G106748, respectively) fell under previously published QTL in one or more mapping populations and are linked to a measurable reduction of aflatoxin in maize grains. Association mapping results found 28 of the 726 SNPs tested were associated with reduced aflatoxin levels at p ≤ 9.71 x 10-4 according to association statistics. These fell within or near nine of the ZmLOX genes. This work confirms the importance of some lipoxygenases for resistance to aflatoxin accumulation and may be used to direct future genetic selection in maize.

Lipoxygenase directed anti-inflammatory and anti-cancerous secondary metabolites: ADMET-based screening, molecular docking and dynamics simulation.

PubMed

Singh, Swati; Awasthi, Manika; Pandey, Veda P; Dwivedi, Upendra N

2017-02-01

Lipoxygenases (LOXs), key enzymes involved in the biosynthesis of leukotrienes, are well known to participate in the inflammatory and immune responses. With the recent reports of involvement of 5-LOX (one of the isozymes of LOX in human) in cancer, there is a need to find out selective inhibitors of 5-LOX for their therapeutic application. In the present study, plant-derived 300 anti-inflammatory and anti-cancerous secondary metabolites (100 each of alkaloids, flavonoids and terpenoids) have been screened for their pharmacokinetic properties and subsequently docked for identification of potent inhibitors of 5-LOX. Pharmacokinetic analyses revealed that only 18 alkaloids, 26 flavonoids, and 9 terpenoids were found to fulfill all the absorption, distribution, metabolism, excretion, and toxicity descriptors as well as those of Lipinski's Rule of Five. Docking analyses of pharmacokinetically screened metabolites and their comparison with a known inhibitor (drug), namely zileuton revealed that only three alkaloids, six flavonoids and three terpenoids were found to dock successfully with 5-LOX with the flavonoid, velutin being the most potent inhibitor among all. The results of the docking analyses were further validated by performing molecular dynamics simulation and binding energy calculations for the complexes of 5-LOX with velutin, galangin, chrysin (in order of LibDock scores), and zileuton. The data revealed stabilization of all the complexes within 15 ns of simulation with velutin complex exhibiting least root-mean-square deviation value (.285 ± .007 nm) as well as least binding energy (ΔG bind  = -203.169 kJ/mol) as compared to others during the stabilization phase of simulation.

Resuscitation of Preterm Neonates With Limited Versus High Oxygen Strategy

PubMed Central

Kapadia, Vishal S.; Chalak, Lina F.; Sparks, John E.; Allen, James R.; Savani, Rashmin C.

2013-01-01

OBJECTIVE: To determine whether a limited oxygen strategy (LOX) versus a high oxygen strategy (HOX) during delivery room resuscitation decreases oxidative stress in preterm neonates. METHODS: A randomized trial of neonates of 24 to 34 weeks’ gestational age (GA) who received resuscitation was performed. LOX neonates received room air as the initial resuscitation gas, and fraction of inspired oxygen (Fio2) was adjusted by 10% every 30 seconds to achieve target preductal oxygen saturations (Spo2) as described by the 2010 Neonatal Resuscitation Program guidelines. HOX neonates received 100% O2 as initial resuscitation gas, and Fio2 was adjusted by 10% to keep preductal Spo2 at 85% to 94%. Total hydroperoxide (TH), biological antioxidant potential (BAP), and the oxidative balance ratio (BAP/TH) were analyzed in cord blood and the first hour of life. Secondary outcomes included delivery room interventions, respiratory support on NICU admission, and short-term morbidities. RESULTS: Forty-four LOX (GA: 30 ± 3 weeks; birth weight: 1678 ± 634 g) and 44 HOX (GA: 30 ± 3 weeks; birth weight: 1463 ± 606 g) neonates were included. LOX decreased integrated excess oxygen (∑Fio2 × time [min]) in the delivery room compared with HOX (401 ± 151 vs 662 ± 249; P < .01). At 1 hour of life, BAP/TH was 60% higher for LOX versus HOX neonates (13 [9–16] vs 8 [6–9]) µM/U.CARR, P < .01). LOX decreased ventilator days (3 [0–64] vs 8 [0–96]; P < .05) and reduced the incidence of bronchopulmonary dysplasia (7% vs 25%; P < .05). CONCLUSIONS: LOX is feasible and results in less oxygen exposure, lower oxidative stress, and decreased respiratory morbidities and thus is a reasonable alternative for resuscitation of preterm neonates in the delivery room. PMID:24218465

Estrogen Alters Remodeling of the Vaginal Wall after Surgical Injury in Guinea Pigs1

PubMed Central

Balgobin, Sunil; Montoya, T. Ignacio; Shi, Haolin; Acevedo, Jesus F.; Keller, Patrick W.; Riegel, Matthew; Wai, Clifford Y.; Word, Ruth Ann

2013-01-01

ABSTRACT Loss of pelvic organ support (i.e., pelvic organ prolapse) is common in menopausal women. Surgical reconstruction of pelvic organ prolapse is plagued with high failure rates. The objective of this study was to determine the effects of estrogen on biomechanical properties, lysyl oxidase (LOX), collagen content, and histomorphology of the vagina with or without surgical injury. Nulliparous ovariectomized guinea pigs were treated systemically with either 50 μg/kg/day estradiol (E2,) or vehicle. After 2 wk, vaginal surgery was performed, and animals were treated with either beta-aminopropionitrile (BAPN, an irreversible LOX inhibitor), or vehicle to determine the role of LOX in recovery of the vaginal wall from injury with or without E2. Estradiol resulted in (i) significant growth, increased smooth muscle, and increased thickness of the vagina, (ii) increased distensibility without compromise of maximal force at failure, and (iii) increased total and cross-linked collagen. In the absence of E2, BAPN resulted in decreased collagen and vaginal wall strength in the area of the injury. In contrast, in E2-treated animals, increased distensibility, maximal forces, and total collagen were maintained despite BAPN. Interestingly, LOX mRNA was induced dramatically (9.5-fold) in the injured vagina with or without E2 at 4 days. By 21 days, however, LOX levels declined to near baseline in E2-deprived animals. LOX mRNA levels remained strikingly elevated (12-fold) at 21 days in the estrogenized vagina. The results suggest that prolonged E2 induced increases in LOX, and collagen cross-links may act to sustain a matrix environment that optimizes long-term surgical wound healing in the vagina. PMID:24174572

A free-piston Stirling engine/linear alternator controls and load interaction test facility

NASA Technical Reports Server (NTRS)

Rauch, Jeffrey S.; Kankam, M. David; Santiago, Walter; Madi, Frank J.

1992-01-01

A test facility at LeRC was assembled for evaluating free-piston Stirling engine/linear alternator control options, and interaction with various electrical loads. This facility is based on a 'SPIKE' engine/alternator. The engine/alternator, a multi-purpose load system, a digital computer based load and facility control, and a data acquisition system with both steady-periodic and transient capability are described. Preliminary steady-periodic results are included for several operating modes of a digital AC parasitic load control. Preliminary results on the transient response to switching a resistive AC user load are discussed.

Lipoxygenase pathways in Homo neanderthalensis: functional comparison with Homo sapiens isoforms

PubMed Central

Chaitidis, Pavlos; Adel, Susan; Anton, Monika; Heydeck, Dagmar; Kuhn, Hartmut; Horn, Thomas

2013-01-01

Lipoxygenases (LOX) have been implicated in biosynthesis of pro- and anti-inflammatory mediators, and a previous report suggested compromised leukotriene signaling in H. neanderthalensis. To search for corresponding differences in leukotriene biosynthesis, we screened the Neandertal genome for LOX genes and found that, as modern humans, this archaic hominid contains six LOX genes (nALOX15, nALOX12, nALOX5, nALOX15B, nALOX12B, and nALOXE3) and one pseudogene. In the Neandertal genome, 60–75% of the amino acids of the different human LOX isoforms have been identified, and the degree of identity varies between 96 and 99%. Most functional amino acids (iron ligands, specificity determinants, calcium and ATP-binding sites, membrane-binding determinants, and phosphorylation sites) are well conserved in the Neandertal LOX isoforms, and expression of selected neandertalized human LOX mutants revealed no major functional defects. However, in nALOX12 and nALOXE3, two premature stop codons were found, leading to inactive enzyme species. These data suggest that ALOX15, ALOX5, ALOX15B, and ALOX12B should have been present as functional enzymes in H. neanderthalensis and that in contrast to lower nonhuman primates (M. mulatta) and other mammals (mice, rats), this ancient hominid expressed a 15-lipoxygenating ALOX15. Expression of ALOXE3 and ALOX12 was compromised, which might have caused problems in epidermal differentiation. PMID:23475662

Development of laboratory testing facility for evaluation of base-soil behavior under repeated loading : phase-1 : feasibility study.

DOT National Transportation Integrated Search

2008-03-01

The main objective of this study was to determine the most beneficial and cost-effective accelerated load facility that can be used in conjunction with LTRCs Accelerated Load Facility (ALF). The facility will be used primarily for conducting preli...

Fundamental Boiling and RP-1 Freezing Experiments

NASA Technical Reports Server (NTRS)

Goode, Brian; Turner, Larry D. (Technical Monitor)

2001-01-01

This paper describes results from experiments performed to help understand certain aspects of the MC-1 engine prestart thermal conditioning procedure. The procedure was constrained by the fact that the engine must chill long enough to get quality LOX at the LOX pump inlet but must be short enough to prevent freezing of RP-1 in the fuel pump. A chill test of an MC-1 LOX impeller was performed in LN2 to obtain data on film boiling, transition boiling and impeller temperature histories. The transition boiling data was important to the chill time so a subsequent experiment was performed chilling simple steel plates in LOX to obtain similar data for LOX. To address the fuel freezing concern, two experiments were performed. First, fuel was frozen in a tray and its physical characteristics were observed and temperatures of the fuel were measured. The result was physical characteristics as a function of temperature. Second was an attempt to measure the frozen thickness of RP-1 on a cold wall submerged in warm RP-1 and to develop a method for calculating that thickness for other conditions.

Design and performance evaluations of a LO2/methane reaction control engine

NASA Astrophysics Data System (ADS)

Johnson, Aaron

Liquid oxygen (LOX) and liquid methane (LCH4) are a propellant combination viewed as a potential enabling technology for spacecraft propulsion. Reasons why LOX/LCH4 is being used as an alternative propellant source include: it is less toxic than other propellants, it has the possibility to be harvested on extraterrestrial soil, LCH4 has a higher energy density than liquid hydrogen (LH2; commonly used on vehicle main engines), and LOX/LCH4 has comparable performance to other well-known propellant combinations. Through the continued partnership between the National Aeronautics and Space Administration (NASA) and the University of Texas at El Paso (UTEP) a LOX/LCH4 reaction control engine (RCE) was developed and researched. The RCE was developed for the purpose of being integrated into two UTEP LOX/LCH4 vehicles, Janus and Daedalus, and was designed based on previous engines tested both at NASA and the center for space exploration and technology research (cSETR) lab. This report details the design process and manufacturing of the engine, cold flow studies evaluating injector design, and preliminary hot fire tests to give insight into engine performance.

The next generation rocket engines

NASA Astrophysics Data System (ADS)

Beichel, Rudi; O'Brien, Charles J.; Taylor, James P.

This paper examines propulsion system technologies for earth-to-orbit vehicles, and describes several propulsion system concepts which could support the recommendations of the Commission for Space Development for the year 2000. The hallmark of that system must and will be reliability. Reliability will be obtained through a very structured design approach, coupled with a rational, cost effective, development and qualification program. To improve the next generation space transportation propulsion systems we need to select the very best of alternative power and performance cycles and engine physical concepts with a rigid requirement to achieve a robust, dependable, affordable propulsion system. For example, engine concepts using either propellants or non-propellant fluids for cooling and/or power drive offer the potential to provide smooth, controlled engine starts, low turbine temperatures, etc. as required for long life turbomachinery. Concepts examined are LOX/LH 2, |LOX/LH 2 + hydrocarbon, and LOX/LH 2 + hydrocarbon + Al dual expander engines, separate LOX/LH 2 and LOX/hydrocarbon engines, and variable mixture ratio engines. A fully reusable propulsion system that is perceived to be very low risk and low in operation cost is described.

Design of a 2000 lbf LOX/LCH4 Throttleable Rocket Engine for a Vertical Lander

NASA Astrophysics Data System (ADS)

Lopez, Israel

Liquid oxygen (LOX) and liquid methane (LCH4) has been recognized as an attractive rocket propellant combination because of its in-situ resource utilization (ISRU) capabilities, namely in Mars. ISRU would allow launch vehicles to carry greater payloads and promote missions to Mars. This has led to an increasing interest to develop spacecraft technologies that employ this propellant combination. The UTEP Center for Space Exploration and Technology Research (cSETR) has focused part of its research efforts to developing LOX/LCH4 systems. One of those projects includes the development of a vertical takeoff and landing vehicle called JANUS. This vehicle will employ a LOX/LCH 4 propulsion system. The main propulsion engine is called CROME-X and is currently being developed as part of this project. This rocket engine will employ LOX/LCH4 propellants and is intended to operate from 2000-500 lbf thrust range. This thesis describes the design and development of CROME-X. Specifically, it describes the design process for the main engine components, the design criteria for each, and plans for future engine development.

Dithiocarbamates are teratogenic to developing zebrafish through inhibition of lysyl oxidase activity

DOE Office of Scientific and Technical Information (OSTI.GOV)

Boxtel, Antonius L. van, E-mail: thijs.van.boxtel@ivm.vu.n; Kamstra, Jorke H.; Fluitsma, Donna M.

2010-04-15

Dithiocarbamates (DTCs) are a class of compounds that are extensively used in agriculture as pesticides. As such, humans and wildlife are undoubtedly exposed to these chemicals. Although DTCs are thought to be relatively safe due to their short half lives, it is well established that they are teratogenic to vertebrates, especially to fish. In zebrafish, these teratogenic effects are characterized by distorted notochord development and shortened anterior to posterior axis. DTCs are known copper (Cu) chelators but this does not fully explain the observed teratogenic effects. We show here that DTCs cause malformations in zebrafish that highly resemble teratogenic effectsmore » observed by direct inhibition of a group of cuproenzymes termed lysyl oxidases (LOX). Additionally, we demonstrate that partial knockdown of three LOX genes, lox, loxl1 and loxl5b, sensitizes the developing embryo to DTC exposure. Finally, we show that DTCs directly inhibit zebrafish LOX activity in an ex vivo amine oxidase assay. Taken together, these results provide the first evidence that DTC induced teratogenic effects are, at least in part, caused by direct inhibition of LOX activity.« less

A proof for substitution of endogenous iron (II) in lipoxygenase by exogenous Cu2+.

PubMed

Cai, Yan; Xu, Hong; Xia, Yongmei; Su, Yafen; Fang, Yun

2010-10-01

Soybean lipoxygenase (LOX) contains endogenous iron (II) at the active site, which is important for the enzyme activity. The activity of LOX can be accelerated by some exogenous metal ions including Cu2+. However, the mechanism of the activity improvement caused by exogenous metal ions remains unclear, not only for LOX but for most other metalloenzymes. Meanwhile, the possibility that exogenous metal ions can displace endogenous iron (II) is still in discussion for a lack of a direct and quantitative proof. In this paper, a quantitative proof of replacing iron (II) inside LOX by exogenous Cu2+ was provided, simply using UV-Vis spectrometry with two indicators p-carboxylantipyrylazo and 9-(4-carboxyphenyl)-2,3,7-trihydroxyl-6-fluorine. A 0.56 microM free iron (II) was observed in the bulk solution after incubating 9.45 microM Cu2+ with 16.10 microM LOX at 20 degrees C for 5 min, which is in coincidence with the decrement of Cu2+ in the bulk solution (0.53 microM), implying that iron (II) was replaced by Cu2+.

Ox-LDL increases OX40L in endothelial cells through a LOX-1-dependent mechanism

PubMed Central

Dong, Q.; Xiang, R.; Zhang, D.Y.; Qin, S.

2013-01-01

Oxidative low-density lipoprotein (Ox-LDL) is a key risk factor for the development of atherosclerosis, and it can stimulate the expression of a variety of inflammatory signals. As a new and highly sensitive inflammation index, OX40L may be a key to understanding the mechanisms that regulate interactions between cells within the vessel wall and inflammatory mediators during the development of atherosclerosis. To investigate whether Ox-LDL regulates OX40L expression through an oxidized LDL-1 receptor (LOX-1)-mediated mechanism, we investigated the effect of different concentrations of Ox-LDL (50, 100, 150 µg/mL) on endothelial cell proliferation and apoptosis. Stimulation with Ox-LDL increased OX40L protein 1.44-fold and mRNA 4.0-fold in endothelial cells, and these effects were inhibited by blocking LOX-1. These results indicate that LOX-1 plays an important role in the chronic inflammatory process in blood vessel walls. Inhibiting LOX-1 may reduce blood vessel inflammation and provide a therapeutic option to limit atherosclerosis progression. PMID:24068192

Combustion of LOX with H2(sub g) under subcritical, critical, and supercritical conditions (Task 1) and experimental observation of dense spray and mixing of impinging jets (Task 2)

NASA Technical Reports Server (NTRS)

Kuo, K. K.; Hsieh, W. H.; Cheung, F. B.; Yang, A. S.; Brown, J. J.; Woodward, R. D.; Kline, M. C.; Burch, R. L.

1992-01-01

The objective was to achieve a better understanding of the combustion processes of liquid oxygen and gaseous hydrogen under broad range of pressure covering subcritical, critical, and supercritical conditions. The scope of the experimental work falls into the following areas: (1) design of the overall experimental setup; (2) modification of an existing windowed high pressure chamber; (3) design of the LOX feeding system; (4) provision of the safety features in the test rig design; (5) LOX cleanliness requirements; (6) cold shock testing; (7) implementation of data acquisition systems; (8) preliminary tests for system checkout; (9) modification of LOX feeding system; and (10) evaporation tests. Progress in each area is discussed.

Large Eddy Simulation of Flame-Turbulence Interactions in a LOX-CH4 Shear Coaxial Injector

DTIC Science & Technology

2012-01-01

heat transfer from dense to light fluids.A previous study on LOX/H2 flames39,40 have pointed the limitations of central scheme to predict such large...pp. 151–169. 39Masquelet, M., Simulations of a Sub-scale Liquid Rocket Engine: Transient Heat Transfer in a Real Gas Environment , Master’s thesis...Eddy Simulation of a cryogenic flame issued from a LOX-CH4 shear coaxial injector. The operating pressure is above the critical pressure for both

Virtual Dual inhibition of COX-2 / 5-LOX enzymes based on binding properties of alpha-amyrins, the anti-inflammatory compound as a promising anti-cancer drug.

PubMed

Ranjbar, Mohammad Mehdi; Assadolahi, Vahideh; Yazdani, Mohsen; Nikaein, Donya; Rashidieh, Behnam

2016-01-01

Hydro-alcoholic fruit extract of Cordia myxa was considerably effective on curing acute inflammation in mouse model. Previous studies suggested significant anti-inflammatory activities as well as potential anticancer agent of α-amyrins in seeds. Inhibition of Cyclooxygenase-2 (COX-2) and 5-Lipooxygenase (5-LOX) is significant in cancer prevention and therapeutics although this inhibition with chemo-drugs has its own side-effects. It is shown that these enzymes pathways are related to several cancers including colon, breast and lung cancer. This study was conducted based on Cordia species' α-amyrins as a safer natural anti-cancer compound for inhibition of COX-2 and 5-LOX enzymes by molecular docking. The X-ray crystal structure of COX2 / 5-LOX enzymes and α-amyrins was retrieved and energetically minimized respectively. The binding site and surface of enzymes were detected. Docking studies were performed by AutoDock 4.2 using Lamarckian genetic algorithm (LGA). Finally drug likeness, molecular pharmacokinetic properties and toxicity of α-amyrins was calculated. Molecular Docking revealed hydrogen and hydrophobic interactions between α-amyrins with both active sites of COX-2 and 5-LOX enzymes. Interestingly, it covalently bonded to Fe cofactor of 5-LOX enzyme and chelated this molecule. Base on binding energies (∆G) α-amyrin has more inhibitory effects on 5-LOX (-10.45 Kcal/mol) than COX-2 (-8.02 Kcal/mol). Analysis of molecular pharmacokinetic parameters suggested that α-amyrins complied with most sets of Lipinski's rules, and so it could be a suitable ligand for docking studies. Eventually, bioactivity score showed α-amyrins possess considerable biological activities as nuclear receptor, enzyme inhibitor, GPCR and protease inhibitor ligand. These results clearly demonstrate that α-amyrins could act as potential highly selective COX-/5-LOX inhibitor. Also, it is a safe compound in comparison with classical non-steroidal anti-inflammatory drugs (NSAIDs) that are known as cancer preventive agents, since it is free of side effects on human body and it can be a promising drug for cancer therapeutics.

Virtual Dual inhibition of COX-2 / 5-LOX enzymes based on binding properties of alpha-amyrins, the anti-inflammatory compound as a promising anti-cancer drug

PubMed Central

Ranjbar, Mohammad Mehdi; Assadolahi, Vahideh; Yazdani, Mohsen; Nikaein, Donya; Rashidieh, Behnam

2016-01-01

Hydro-alcoholic fruit extract of Cordia myxa was considerably effective on curing acute inflammation in mouse model. Previous studies suggested significant anti-inflammatory activities as well as potential anticancer agent of α-amyrins in seeds. Inhibition of Cyclooxygenase-2 (COX-2) and 5-Lipooxygenase (5-LOX) is significant in cancer prevention and therapeutics although this inhibition with chemo-drugs has its own side-effects. It is shown that these enzymes pathways are related to several cancers including colon, breast and lung cancer. This study was conducted based on Cordia species' α-amyrins as a safer natural anti-cancer compound for inhibition of COX-2 and 5-LOX enzymes by molecular docking. The X-ray crystal structure of COX2 / 5-LOX enzymes and α-amyrins was retrieved and energetically minimized respectively. The binding site and surface of enzymes were detected. Docking studies were performed by AutoDock 4.2 using Lamarckian genetic algorithm (LGA). Finally drug likeness, molecular pharmacokinetic properties and toxicity of α-amyrins was calculated. Molecular Docking revealed hydrogen and hydrophobic interactions between α-amyrins with both active sites of COX-2 and 5-LOX enzymes. Interestingly, it covalently bonded to Fe cofactor of 5-LOX enzyme and chelated this molecule. Base on binding energies (∆G) α-amyrin has more inhibitory effects on 5-LOX (-10.45 Kcal/mol) than COX-2 (-8.02 Kcal/mol). Analysis of molecular pharmacokinetic parameters suggested that α-amyrins complied with most sets of Lipinski's rules, and so it could be a suitable ligand for docking studies. Eventually, bioactivity score showed α-amyrins possess considerable biological activities as nuclear receptor, enzyme inhibitor, GPCR and protease inhibitor ligand. These results clearly demonstrate that α-amyrins could act as potential highly selective COX-/5-LOX inhibitor. Also, it is a safe compound in comparison with classical non-steroidal anti-inflammatory drugs (NSAIDs) that are known as cancer preventive agents, since it is free of side effects on human body and it can be a promising drug for cancer therapeutics. PMID:27231478

Development of selectable marker free, insect resistant, transgenic mustard (Brassica juncea) plants using Cre/lox mediated recombination

PubMed Central

2013-01-01

Background Antibiotic/ herbicide resistant marker genes have been proven to be very useful in plant transformation for the initial selection of desired transgenic events. However, presence of these genes in the genetically modified crops may render the crop less acceptable to the consumers. Among several different approaches, the effectiveness of Cre/lox mediated recombination strategy for selectable marker gene (SMG) elimination has previously been demonstrated by different groups in several plants including Brassica. In the present study exploiting Cre/lox mediated recombination strategy, attempt has been made for selectable marker gene elimination from Allium sativum leaf agglutinin (ASAL) expressing Brassica plants with hemipteran insect resistant phenotype. Results Allium sativum leaf agglutinin (ASAL) linked with lox flanked hygromycin resistant (hpt) gene was introduced in mustard. Cre recombinase gene cassette was also integrated in separate event. A Cre/lox mediated recombination using crossing strategy was adopted to remove the hpt gene from the subsequent generation of selected hybrid events. Reciprocal crosses were made between T1ASAL-lox-hpt-lox and cre-bar plants. Marker gene elimination was confirmed in the resulting F1 hybrid progenies by PCR analysis, using hpt, cre and ASAL specific primers followed by Southern hybridization. In marker free plants, expression of ASAL was also confirmed by western blotting and ELISA analysis. Retention of functionality of expressed ASAL was investigated by agglutination assay using rabbit erythrocytes. Expressed ASAL was also found to be thermo-sensitive. In planta insect bioassay on F1 hybrid progenies exhibited detrimental effect on the performance of devastating target pest, Lipaphis erysimi. The F1 hybrid hpt negative, ASAL positive plants were allowed to self- fertilize to obtain F2 progeny plants. In some of these plants cre gene was found to be segregated out of the ASAL gene by genetic segregation yielding completely marker free plants. Conclusions The present study establishes the efficient expression of the newly introduced insect resistant ASAL gene even after Cre/lox mediated recombination resulting in elimination of selectable marker gene. PMID:24144281

Floor Plans Fuel Tank Support, Fuel Platform, and LOX ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Floor Plans - Fuel Tank Support, Fuel Platform, and LOX Platform Plans - Marshall Space Flight Center, F-1 Engine Static Test Stand, On Route 565 between Huntsville and Decatur, Huntsville, Madison County, AL

Sound speed measurements in liquid oxygen-liquid nitrogen mixtures

NASA Technical Reports Server (NTRS)

Zuckerwar, A. J.; Mazel, D. S.

1985-01-01

The sound speed in liquid oxygen (LOX), liquid nitrogen (LN2), and five LOX-LN2 mixtures was measured by an ultrasonic pulse-echo technique at temperatures in the vicinity of -195.8C, the boiling point of N2 at a pressure of I atm. Under these conditions, the measurements yield the following relationship between sound speed in meters per second and LN2 content M in mole percent: c = 1009.05-1.8275M+0.0026507 M squared. The second speeds of 1009.05 m/sec plus or minus 0.25 percent for pure LOX and 852.8 m/sec plus or minus 0.32 percent for pure LN2 are compared with those reported by past investigators. Measurement of sound speed should prove an effective means for monitoring the contamination of LOX by Ln2.

Investigations of human platelet-type 12-lipoxygenase: role of lipoxygenase products in platelet activation1[S

PubMed Central

Ikei, Kenneth N.; Yeung, Jennifer; Apopa, Patrick L.; Ceja, Jesús; Vesci, Joanne; Holinstat, Michael

2012-01-01

Human platelet-type 12-lipoxygenase (12-LOX) has recently been shown to play an important role in regulation of human platelet function by reacting with arachidonic acid (AA). However, a number of other fatty acids are present on the platelet surface that, when cleaved from the phospholipid, can be oxidized by 12-LOX. We sought to characterize the substrate specificity of 12-LOX against six essential fatty acids: AA, dihomo-γ-linolenic acid (DGLA), eicosapentaenoic acid (EPA), α-linolenic acid (ALA), eicosadienoic acid (EDA), and linoleic acid (LA). Three fatty acids were comparable substrates (AA, DGLA, and EPA), one was 5-fold slower (ALA), and two showed no reactivity with 12-LOX (EDA and LA). The bioactive lipid products resulting from 12-LOX oxidation of DGLA, 12-(S)-hydroperoxy-8Z,10E,14Z-eicosatrienoic acid [12(S)-HPETrE], and its reduced product, 12(S)-HETrE, resulted in significant attenuation of agonist-mediated platelet aggregation, granule secretion, αIIbβ3 activation, Rap1 activation, and clot retraction. Treatment with DGLA similarly inhibited PAR1-mediated platelet activation as well as platelet clot retraction. These observations are in surprising contrast to our recent work showing 12(S)-HETE is a prothrombotic bioactive lipid and support our hypothesis that the overall effect of 12-LOX oxidation of fatty acids in the platelet is dependent on the fatty acid substrates available at the platelet membrane. PMID:22984144

Platelet 12-lipoxygenase activation via glycoprotein VI: involvement of multiple signaling pathways in agonist control of H(P)ETE synthesis.

PubMed

Coffey, Marcus J; Jarvis, Gavin E; Gibbins, Jonathan M; Coles, Barbara; Barrett, Natasha E; Wylie, Oliver R E; O'Donnell, Valerie B

2004-06-25

Lipoxygenases (LOX) contribute to vascular disease and inflammation through generation of bioactive lipids, including 12-hydro(pero)xyeicosatetraenoic acid (12-H(P)ETE). The physiological mechanisms that acutely control LOX product generation in mammalian cells are uncharacterized. Human platelets that contain a 12-LOX isoform (p12-LOX) were used to define pathways that activate H(P)ETE synthesis in the vasculature. Collagen and collagen-related peptide (CRP) (1 to 10 microg/mL) acutely induced platelet 12-H(P)ETE synthesis. This implicated the collagen receptor glycoprotein VI (GPVI), which signals via the immunoreceptor-based activatory motif (ITAM)-containing FcRgamma chain. Conversely, thrombin only activated at high concentrations (> 0.2 U/mL), whereas U46619 and ADP alone were ineffective. Collagen or CRP-stimulated 12-H(P)ETE generation was inhibited by staurosporine, PP2, wortmannin, BAPTA/AM, EGTA, and L-655238, implicating src-tyrosine kinases, PI3-kinase, Ca2+ mobilization, and p12-LOX translocation. In contrast, protein kinase C (PKC) inhibition potentiated 12-H(P)ETE generation. Finally, activation of the immunoreceptor tyrosine-based inhibitory motif (ITIM)-containing platelet endothelial cell adhesion molecule (PECAM-1) inhibited p12-LOX product generation. This study characterizes a receptor-dependent pathway for 12-H(P)ETE synthesis via the collagen receptor GPVI, which is negatively regulated by PECAM-1 and PKC, and demonstrates a novel link between immune receptor signaling and lipid mediator generation in the vasculature.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Saad, Fawzy A.; Harvard Medical School, Boston, MA 02115; Torres, Marie

LOX, the principal enzyme involved in crosslinking of collagen, was the first of several lysyl oxidase isotypes to be characterized. Its active form was believed to be exclusively extracellular. Active LOX was later reported to be present in cell nuclei; its function there is unknown. LOX expression opposes the effect of mutationally activated Ras, which is present in about 30% of human cancers. The mechanism of LOX in countering the action of Ras is also unknown. In the present work, assessment of nuclear protein for possible effects of lysyl oxidase activity led to the discovery that proliferating cells dramatically increasemore » their nuclear protein content when exposed to BAPN ({beta}-aminopropionitrile), a highly specific lysyl oxidase inhibitor that reportedly blocks LOX inhibition of Ras-induced oocyte maturation. In three cell types (PC12 cells, A7r5 smooth muscle cells, and NIH 3T3 fibroblasts), BAPN caused a 1.8-, 1.7-, and 2.1-fold increase in total nuclear protein per cell, respectively, affecting all major components in both nuclear matrix and chromatin fractions. Since nuclear size is correlated with proliferative status, enzyme activity restricting nuclear growth may be involved in the lysyl oxidase tumor suppressive effect. Evidence is also presented for the presence of apparent lysyl oxidase isotype(s) containing a highly conserved LOX active site sequence in the nuclei of PC12 cells, which do not manufacture extracellular lysyl oxidase substrates. Results reported here support the hypothesis that nuclear lysyl oxidase regulates nuclear growth, and thereby modulates cell proliferation.« less

Ignition Characterization Test Results for the LO2/Ethanol Propellant Combination

NASA Technical Reports Server (NTRS)

Robinson, Philip J.; Popp, Christopher G.; veith, Eric M.

2007-01-01

A series of contracts were issued by the Marshall Space Flight Center (MSFC) of the National Aeronautics and Space Administration (NASA) un der the auspices of the Exploration Systems Mission Directorate to de velop and expand the maturity of candidate technologies considered to be important for future space exploration. One such technology was to determine the viability of incorporating non-toxic propellants for R eaction Control Subsystems (RCS). Contract NAS8-01109 was issued to A erojet to develop a dual thrust Reaction Control Engine (RCE) that ut ilized liquid oxygen and ethanol as the propellants. The dual thrust RCE incorporated a primary thrust level of 870 lbf, and a vernier thru st level of 10 - 30 lbf. The preferred RCS approach for the dual thru st RCE was to utilize pressure-fed liquid oxygen (LOX) and ethanol pr opellants; however, previous dual thrust feasibility testing incorporated GOX/Ethanol igniters as opposed to LOX/Ethanol igniters in the de sign. GOX/Ethanol was easier to ignite, but this combination had syst em design implications of providing GOX for the igniters. A LOX/Ethan ol igniter was desired; however, extensive LOX/Ethanol ignition data over the anticipated operating range for the dual thrust RCE did not e xist. Therefore, Aerojet designed and tested a workhorse LOX igniter to determine LOX/Ethanol ignition characteristics as part of a risk m itigation effort for the dual thrust RCE design. The objective of the ignition testing was to demonstrate successful ignition from GOX to LOX, encompassing potential two-phase flow conditions anticipated being present in real mission applications. A workhorse igniter was desig ned to accommodate the full LOX design flowrate, as well as a reduced GOX flowrate. It was reasoned that the initial LOX flow through the igniter would flash to GOX due to the latent heat stored in the hardwa re, causing a reduced oxygen flowrate because of a choked, or sonic, flow condition through the injection elements. As LOX flow continued, the hardware would chill-in, with the injected oxygen flow transitioning from cold GOX through two'phase flow to subcooled LOX. The Workh orse igniter was well instrumented: Pressure and temperature instrumentation permitted oxygen state points to be determined in the igniter oxidizer manifold, and gas-side igniter chamber thermocouples provide d chamber thermal profile characteristics. The cold flow chamber pres sure (Pc) for each test was determined and coupled with the igniter chamber diameter (De) to calculate the characteristic quench parameter (Pc x Dc), which was plotted as a function of core mixture ratio, MRc . Ignition limits were determined over a broad range of valve inlet conditions, and ignition was demonstrated with oxygen inlet conditions that ranged from subcooled 210 deg R LOX to 486 deg R GOX. Once ign ited at cold GOX conditions, combustion was continuous as the hardwar e chilled in and the core mixture ratio transitioned from values near 1.0 to over 12.5. Pulsing is required in typical RCS engines; therefore, the workhorse igniter was pulse tested to verify the ability to pr ovide the required ignition for a pulsing RCE. The minimum electrical pulse width (EPW) of the dual thrust RCE was 0.080 seconds. Igniter pulse tests were performed at three conditions: (1) an EPW of 0.080 se conds at 25% duty cycle for 400 pulses; (2) an EPW of 0.160 seconds a nd a 5% duty cycle for 124 pulses; (3) an EPW of 0.160 seconds and a 50% duty cycle for 380 pulses. Successful ignition of LOX/Ethanol was demonstrated over a broad range of valve inlet conditions, with the empirically determined LOX/Ethanol ignition limits extending the previous database established for GOX/Ethanol ignition limits. Although th e observed chill-in characteristics of the hardware varied significan tly with flowrate, ignition was readily achieved. Combustion was marg inal at extremely fuel-rich conditions, and it fluctuated as the oxygen passed rough the twophase flow regime during the period of hardware chill-in. Pulse testing showed good repeatability with 100 percent r e-ignition for all pulses. Certain pulse-to-pulse repeatability requirements for actual RCS operation may necessitate establishment of mini mum oxygen flow rates and engine thrust levels for satisfactory engin e performance.

Lipid-modifying enzymes in oat and faba bean.

PubMed

Yang, Zhen; Piironen, Vieno; Lampi, Anna-Maija

2017-10-01

The aim was to study lipase, lipoxygenase (LOX) and peroxygenase (POX) activities in oat and faba bean samples to be able to evaluate their potential in formation of lipid-derived off-flavours. Lipase and LOX activities were measured by spectroscopy, and POX activities via the formation of epoxides. An ultra-high performance liquid chromatography method was developed to study the formation of fatty acid epoxides. The epoxides of esters were measured by gas chromatography. Mass spectroscopy was used to verify the identity of the epoxides. Both oat and faba bean possessed high lipase activities. In faba bean, LOX catalysed the formation of hydroperoxides, whose break-down products are the likely cause of off-flavours. Since oat had low LOX activity, autoxidation is needed to initiate lipid oxidation. Oat had high POX activity, which is able to convert hydroperoxides to epoxy and hydroxy fatty acids that could contribute significantly to off-flavours. POX activity in the faba bean was low. Thus, in faba bean volatile lipid oxidation products could rapidly be formed by LOX, whereas in oat reactions are slower due to the need of autoxidation prior to further reactions. Copyright © 2017 Elsevier Ltd. All rights reserved.

Inhibition of 12/15 lipoxygenase by curcumin and an extract from Curcuma longa L.

PubMed

Bezáková, Lýdia; Košťálová, Daniela; Obložinský, Marek; Hoffman, Peter; Pekárová, Mária; Kollárová, Renáta; Holková, Ivana; Mošovská, Silvia; Sturdík, Ernest

2014-02-01

Curcumin (diferuloylmethane) is an orange-yellow secondary metabolic compound from the rhizome of turmeric (Curcuma longa L.), a spice often found in curry powder. It is one of the major curcuminoids of turmeric. For centuries, curcumin has been used in some medicinal preparations or as a food colouring agent. A variety of enzymes that are closely associated with inflammation and cancer were found to be modulated by curcumin. This paper summarized the results of the inhibitory effect of curcumin and a Curcuma longa L. ethanolic extract on lipoxygenase from the rat lung cytosolic fraction. The positional specificity determination of arachidonic acid dioxygenation by RP- and SP-HPLC methods showed that in a purified enzyme preparation from the rat lung cytosol the specific form of lipoxygenase (LOX) is present exhibiting 12/15-LOX dual specificity (with predominant 15-LOX activity). The inhibitory activity of curcumin and Curcuma longa extract on LOX from cytosolic fraction of rat lung was expressed in the percentage of inhibition and as IC50. Lineweaver-Burk plot analysis has indicated that curcumin is the competitive inhibitor of 12/15 LOX from the rat lung cytosolic fraction.

In vitro kinetics of soybean lipoxygenase with combinatorial fatty substrates and its functional significance in off flavour development.

PubMed

Mandal, Somnath; Dahuja, Anil; Kar, Abhijit; Santha, I M

2014-03-01

Lipoxygenase (Lox) mediated oxidation of polyunsaturated fatty acids (PUFA) in mature soya seeds results in objectionable flavour. In the present study, Lox isozymes were purified to near homogeneity (107-fold). Lox-2 and 3 displayed remarkable kinetic preference (1.7 and 1.5-fold, respectively) for low PUFA ratios (LA/LeA) (PRs) among the selected PUFA combinations. Lox-1 displayed no specific preference. Pure Lox-1 displayed unbiased response towards substrates with marginal preference (1.2-fold) for linoleic acid at its optimum pH. Volatile compounds profiling showed a direct relationship between PRs and hexanal to trans-2-hexenal (1.47, 2.24 and 18.90 for 2, 7 and 15 PRs, respectively) ratio. The off-flavour determining parameters like TBA value, carbonyl value and lipid hydroperoxides (LHPODs) exhibited significant negative correlation (0.76, 0.74, 0.72; p<0.0001) in selected soya genotypes displaying varied PRs and significant positive correlation (0.89, 0.81. 0.89; p<0.0001) with ratio of PI (polyene index) to PRs - suggesting the plausible significance of PUFA ratios in biological lipid peroxidation. Copyright © 2013 Elsevier Ltd. All rights reserved.

Agrobacterium-mediated transformation of maize (Zea mays) with Cre-lox site specific recombination cassettes in BIBAC vectors.

PubMed

Vega, Juan M; Yu, Weichang; Han, Fangpu; Kato, Akio; Peters, Eric M; Zhang, Zhanyuan J; Birchler, James A

2008-04-01

The Cre/loxP site-specific recombination system has been applied in various plant species including maize (Zea mays) for marker gene removal, gene targeting, and functional genomics. A BIBAC vector system was adapted for maize transformation with a large fragment of genetic material including a herbicide resistance marker gene, a 30 kb yeast genomic fragment as a marker for fluorescence in situ hybridization (FISH), and a 35S-lox-cre recombination cassette. Seventy-five transgenic lines were generated from Agrobacterium-mediated transformation of a maize Hi II line with multiple B chromosomes. Eighty-four inserts have been localized among all 10 A chromosome pairs by FISH using the yeast DNA probe together with a karyotyping cocktail. No inserts were found on the B chromosomes; thus a bias against the B chromosomes by the Agrobacterium-mediated transformation was revealed. The expression of a cre gene was confirmed in 68 of the 75 transgenic lines by a reporter construct for cre/lox mediated recombination. The placement of the cre/lox site-specific recombination system in many locations in the maize genome will be valuable materials for gene targeting and chromosome engineering.

High-pressure LOX/CH4 injector program

NASA Technical Reports Server (NTRS)

Wheeler, D. B.; Kirby, F. M.

1979-01-01

Two injector types, either coaxial or impinging elements, for high pressure LOX/CH4 operation with an existing 40K chamber are examined. A comparison is presented. The detailed fabrication drawings and supporting analysis are presented.

Cycom 977-2 Composite Material: Impact Test Results (workshop presentation)

NASA Technical Reports Server (NTRS)

Engle, Carl; Herald, Stephen; Watkins, Casey

2005-01-01

Contents include the following: Ambient (13A) tests of Cycom 977-2 impact characteristics by the Brucenton and statistical method at MSFC and WSTF. Repeat (13A) tests of tested Cycom from phase I at MSFC to expended testing statistical database. Conduct high-pressure tests (13B) in liquid oxygen (LOX) and GOX at MSFC and WSTF to determine Cycom reaction characteristics and batch effect. Conduct expended ambient (13A) LOX test at MSFC and high-pressure (13B) testing to determine pressure effects in LOX. Expend 13B GOX database.

PgLOX6 encoding a lipoxygenase contributes to jasmonic acid biosynthesis and ginsenoside production in Panax ginseng.

PubMed

Rahimi, Shadi; Kim, Yu-Jin; Sukweenadhi, Johan; Zhang, Dabing; Yang, Deok-Chun

2016-11-01

Ginsenosides, the valuable pharmaceutical compounds in Panax ginseng, are triterpene saponins that occur mainly in ginseng plants. It was shown that in vitro treatment with the phytohormone jasmonic acid (JA) is able to increase ginsenoside production in ginseng plants. To understand the molecular link between JA biosynthesis and ginsenoside biosynthesis, we identified a JA biosynthetic 13-lipoxygenase gene (PgLOX6) in P. ginseng that promotes ginsenoside production. The expression of PgLOX6 was high in vascular bundles, which corresponds with expression of ginsenoside biosynthetic genes. Consistent with the role of PgLOX6 in synthesizing JA and promoting ginsenoside synthesis, transgenic plants overexpressing PgLOX6 in Arabidopsis had increased amounts of JA and methyl jasmonate (MJ), increased expression of triterpene biosynthetic genes such as squalene synthase (AtSS1) and squalene epoxidase (AtSE1), and increased squalene content. Moreover, transgenic ginseng roots overexpressing PgLOX6 had around 1.4-fold increased ginsenoside content and upregulation of ginsenoside biosynthesis-related genes including PgSS1, PgSE1, and dammarenediol synthase (PgDDS), which is similar to that of treatment with MJ. However, MJ treatment of transgenic ginseng significantly enhanced JA and MJ, associated with a 2.8-fold increase of ginsenoside content compared with the non-treated, non-transgenic control plant, which was 1.4 times higher than the MJ treatment effect on non-transgenic plants. These results demonstrate that PgLOX6 is responsible for the biosynthesis of JA and promotion of the production of triterpenoid saponin through up-regulating the expression of ginsenoside biosynthetic genes. This work provides insight into the role of JA in biosynthesizing secondary metabolites and provides a molecular tool for increasing ginsenoside production. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Stability-indicating chromatographic methods for the determination of some oxicams.

PubMed

Taha, Elham Anwer; Salama, Nahla Nour; Abdel Fattah, Laila el-Said

2004-01-01

Two sensitive and selective methods were developed for the determination of some oxicams, namely, lornoxicam (LOX), tenoxicam (TEX), and meloxicam (MEX), in the presence of their alkaline degradation products. The first method is based on the thin-layer chromatographic separation of the 3 drugs from their alkaline degradation products, followed by densitometric measurement of the intact drug spots for LOX, TEX, and MEX at 380, 370, and 364 nm, respectively. The developing systems used for separation are ethyl acetate-methanol-26% ammonia (17 + 3 + 0.35, v/v/v) for LOX and TEX and chloroform-n-hexane-96.0% acetic acid (18 + 1 + 1, v/v/v) for MEX. The linear ranges were 0.25-6.0 microg/spot for LOX and TEX and 0.5-10 microg/spot for MEX, with mean recoveries of 99.80 +/- 1.32, 100.57 +/- 1.34, and 100.71 +/- 1.57%, respectively. The second method is based on the liquid chromatographic separation of the 3 drugs from their alkaline degradation products on a reversed-phase C18 column, using mobile phases of methanol-acetonitrile-acetate buffer, pH 4.6 (4.5 + 0.5 + 5.0, v/v/v) for LOX and MEX and methanol-acetonitrile-acetate buffer, pH 4.6 (1.9 + 0.1 + 3.0, v/v/v) for TEX at ambient temperature. Quantification is achieved by UV detection at 280 nm, based on peak area. The linear ranges were 0.5-20 microg/mL for LOX and TEX and 1.25-50 microg/mL for MEX, with mean recoveries of 99.81 +/- 1.01, 98.90 +/- 1.61, and 100.86 +/- 1.55%, respectively. The methods were validated according to guidelines of the International Conference on Harmonization. The developed methods were successfully applied to the determination of LOX, TEX, and MEX in bulk powder, laboratory-prepared mixtures containing different percentages of degradation products, and pharmaceutical dosage forms.

Evaluation of adherence to ambulatory liquid oxygen treatment: are commercialized dual-pressure transducers helpful?

PubMed

van Zeller, Mafalda; Eusébio, Ermelinda; Almeida, João; Winck, João Carlos

2014-09-01

Treatment adherence is widely recognized as a critical problem in long-term oxygen therapy, particularly in ambulatory liquid oxygen (LOX) systems. Adherence-monitoring strategies may be helpful in managing patients. We evaluated subjects' adherence to LOX using VisionOx and compared these results with the subjects' own adherence diaries and self-reported perceptions of use. Patients using LOX were recruited for a clinical interview; the number of days/week and the mean time of use according to subjects' perceptions were recorded. A 14-day diary was provided for every subject while VisionOx was attached to the LOX. VisionOx is a small device that uses pressure transducers to detect oxygen flow and the subject's breathing frequency. Information is stored and downloaded using dedicated software. Nineteen subjects were included (57.9% male with a median age of 63 years). When asked about the perception of LOX use, subjects self-reported using the device for a median of 100.0% of days (78.9% reported to have used it every day) for a median time of 180 min/day. According to data from VisionOx and subjects' diaries during the 14-day evaluation period, the median use was 92.8% of days for 210 min/day. No difference was found between the diaries and VisionOx data. Regarding subjects' perceptions of use, the declared use of LOX percent was significantly higher than reported in the diaries (P = .045) and VisionOx monitoring (P = .045) even though both underestimated the median use per day. Subjects overestimated adherence to LOX therapy (when measuring percent of days of use) compared to adherence diary and objective adherence monitoring. Because no significant difference was found comparing the diaries and VisionOx use, either may be helpful in clinical practice. Copyright © 2014 by Daedalus Enterprises.

Ginkgolide B Reduces LOX-1 Expression by Inhibiting Akt Phosphorylation and Increasing Sirt1 Expression in Oxidized LDL-Stimulated Human Umbilical Vein Endothelial Cells

PubMed Central

Chen, Beidong; Li, Xingguang; Qi, Ruomei

2013-01-01

Oxidized low-density lipoprotein (ox-LDL) is an important risk factor in the development of atherosclerosis. LOX-1, a lectin-like receptor for ox-LDL, is present primarily on endothelial cells and upregulated by ox-LDL, tumor necrosis factor a, shear stress, and cytokines in atherosclerosis. Recent studies demonstrated that ginkgolide B, a platelet-activating factor receptor antagonist, has antiinflammatory and antioxidant effects on endothelial and nerve cells. The present study investigated the effects of ginkgolide B on LOX-1 expression and the possible mechanism of action. Our results showed that ginkgolide B inhibited LOX-1 and intercellular cell adhesion molecule-1 (ICAM-1) expression in ox-LDL-stimulated endothelial cells through a mechanism associated with the attenuation of Akt activation. Similar data were obtained by silencing Akt and LY294002. We also evaluated Sirt1 and nuclear factor erythroid 2-related factor 2 (Nrf2) expression. These molecules play a protective role in endothelial cell injury. The results showed that ginkgolide B increased Sirt1 expression in ox-LDL-treated cells. The inhibitory effects of ginkgolide B on LOX-1 and ICAM-1 expression were reduced in Sirt1 siRNA-transfected cells. Nrf2 expression was increased in ox-LDL-treated cells, and ginkgolide B downregulated Nrf2 expression. These results suggest that ginkgolide B reduces Nrf2 expression by inhibiting LOX-1 expression, consequently reducing oxidative stress injury in ox-LDL-stimulated cells. Altogether, these results indicate that the protective effect of ginkgolide B on endothelial cells may be attributable to a decrease in LOX-1 expression and an increase in Sirt1 expression in ox-LDL-stimulated endothelial cells, the mechanism of which is linked to the inhibition of Akt activation. Ginkgolide B may be a multiple-target drug that exerts protective effects in ox-LDL-treated human umbilical vein endothelial cells. PMID:24069345

Thermal Analysis of the MC1 Engine Turbopump

NASA Technical Reports Server (NTRS)

Roman, Jose; Turner, Larry D. (Technical Monitor)

2001-01-01

The MC1 Engine turbopump supplied the propellants to the main injector. The turbopump consisted of four parts; lox pump, interpropellant seal package (IPS), RP pump and turbine. The thermal analysis was divided into two 2D finite element models; Housing or stationary parts and rotor or rotating parts. Both models were analyzed at the same boundary conditions using SINDA. The housing model consisted of, lox pump housing, ips housing, RP housing, turbine inlet housing, turbine housing, exit guide vane, heat shield and both bearing outer races. The rotor model consisted of the lox impeller; lox end bearing and id race, RP impeller, and RP bearing and id race, shaft and turbine disk. The objectives of the analysis were to: (1) verified the original design and recommend modifications to it, (2) submitted a thermal environment to support the structural analysis, (3) support the component and engine test program. and (4) to support the X34 vehicle program.

Thermal Analysis of the MCI Engine Turbopump

NASA Technical Reports Server (NTRS)

Roman, Jose

2002-01-01

The MCI Engine turbopump supplied the propellants to the main injector. The turbopump consisted of four parts; lox pump, interpropellant seal package (IPS), RP pump and turbine. The thermal analysis was divided into two 2D finite element models; Housing or stationary parts and rotor or rotating parts. Both models were analyzed at the same boundary conditions using SINDA. The housing model consisted of; lox pump housing, ips housing, RP housing, turbine inlet housing, turbine housing, exit guide vane, heat shield and both bearing outer races. The rotor model consisted of the lox impeller; lox end bearing and id race, RP impeller, and RP bearing and id race, shaft and turbine disk. The objectives of the analysis were to (1) verified the original design and recommend modifications to it, (2) submitted a thermal environment to support the structural analysis, (3) support the component and engine test program and (4) to support the X34 vehicle program.

LOX/Methane Main Engine Igniter Tests and Modeling

NASA Technical Reports Server (NTRS)

Breisacher, Kevin J.; Ajmani, Kumund

2008-01-01

The LOX/methane propellant combination is being considered for the Lunar Surface Access Module ascent main engine propulsion system. The proposed switch from the hypergolic propellants used in the Apollo lunar ascent engine to LOX/methane propellants requires the development of igniters capable of highly reliable performance in a lunar surface environment. An ignition test program was conducted that used an in-house designed LOX/methane spark torch igniter. The testing occurred in Cell 21 of the Research Combustion Laboratory to utilize its altitude capability to simulate a space vacuum environment. Approximately 750 ignition test were performed to evaluate the effects of methane purity, igniter body temperature, spark energy level and frequency, mixture ratio, flowrate, and igniter geometry on the ability to obtain successful ignitions. Ignitions were obtained down to an igniter body temperature of approximately 260 R with a 10 torr back-pressure. The data obtained is also being used to anchor a CFD based igniter model.

A dual inhibitor of cyclooxygenase and 5-lipoxygenase protects against kainic acid-induced brain injury.

PubMed

Minutoli, Letteria; Marini, Herbert; Rinaldi, Mariagrazia; Bitto, Alessandra; Irrera, Natasha; Pizzino, Gabriele; Pallio, Giovanni; Calò, Margherita; Adamo, Elena Bianca; Trichilo, Vincenzo; Interdonato, Monica; Galfo, Federica; Squadrito, Francesco; Altavilla, Domenica

2015-06-01

Systemic administration of kainic acid causes inflammation and apoptosis in the brain, resulting in neuronal loss. Dual cyclooxygenase/5-lipoxygenase (COX/5-LOX) inhibitors could represent a possible neuroprotective approach in preventing glutamate excitotoxicity. Consequently, we investigated the effects of a dual inhibitor of COX/5-LOX following intraperitoneal administration of kainic acid (KA, 10 mg/kg) in rats. Animals were randomized to receive either the dual inhibitor of COX/5-LOX (flavocoxid, 20 mg/kg i.p.) or its vehicle (1 ml/kg i.p.) 30 min after KA administration. Sham brain injury rats were used as controls. We evaluated protein expression of phosphorylated extracellular signal-regulated kinase (p-ERK1/2) and tumor necrosis factor alpha (TNF-α) as well as levels of malondialdehyde (MDA), prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) in the hippocampus. Animals were also observed for monitoring behavioral changes according to Racine Scale. Finally, histological analysis and brain edema evaluation were carried out. Treatment with the dual inhibitor of COX/5-LOX decreased protein expression of p-ERK1/2 and TNF-α in hippocampus, markedly reduced MDA, LTB4 and PGE2 hippocampal levels, and also ameliorated brain edema. Histological analysis showed a reduction in cell damage in rats treated with the dual inhibitor of COX/5-LOX, particularly in hippocampal subregion CA3c. Moreover, flavocoxid significantly improved behavioral signs following kainic acid administration. Our results suggest that dual inhibition of COX/5-LOX by flavocoxid has neuroprotective effects during kainic acid-induced excitotoxicity.

Inhibitor of lysyl oxidase improves cardiac function and the collagen/MMP profile in response to volume overload.

PubMed

El Hajj, Elia C; El Hajj, Milad C; Ninh, Van K; Gardner, Jason D

2018-05-18

The cardiac extracellular matrix is a complex architectural network that serves many functions including providing structural and biochemical support to surrounding cells, and regulating intercellular signaling pathways. Cardiac function is directly affected by extracellular matrix (ECM) composition, and alterations of the ECM contribute to progression of heart failure. Initially, collagen deposition is an adaptive response that aims to preserve tissue integrity and maintain normal ventricular function. However, the synergistic effects of the pro-inflammatory and pro-fibrotic responses induce a vicious cycle which causes excess activation of myofibroblasts, significantly increasing collagen deposition and accumulation in the matrix. Further, excess synthesis and activation of the enzyme lysyl oxidase (LOX) during disease increases collagen cross-linking, which significantly increases collagen resistance to degradation by matrix metalloproteinases (MMPs). In this study, the aortocaval fistula model of volume overload (VO) was used to determine whether LOX inhibition could prevent adverse changes in the ECM and subsequent cardiac dysfunction. The major findings from this study are that LOX inhibition: (a) prevented VO-induced increases in LV wall stress, (b) partially attenuated VO-induced ventricular hypertrophy, (c) completely blocked the increases in fibrotic proteins, including collagens, MMPs, and their tissue inhibitors (TIMPs), and (d) prevented the VO-induced decline in cardiac function. It remains unclear whether a direct interaction between LOX and MMPs exists; however our studies suggest a potential link between the two since LOX inhibition completely attenuated the VO-induced increases in MMPs. Overall, our studies demonstrate key cardioprotective effects of LOX inhibition against adverse cardiac remodeling due to chronic VO.

The copper dependent-lysyl oxidases contribute to the pathogenesis of pulmonary emphysema in chronic obstructive pulmonary disease patients.

PubMed

Besiktepe, Neziha; Kayalar, Ozgecan; Ersen, Ezel; Oztay, Fusun

2017-12-01

Abnormalities in the elastic fiber biology are seen in pulmonary emphysema (PE). The copper-dependent lysyl oxidases regulate the production and accumulation of elastic fibers in the connective tissue. This study focused on the relationship between lysyl oxidase (LOX), LOX-like protein 1 (LOXL1), and LOXL2 and PE pathogenesis. Lung samples with or without PE from patients with chronic obstructive lung disease (n=35) were used. Protein levels of elastin, LOX, LOXL1, LOXL2, hypoxia inducible factor 1-alpha (HIF-1α), copper metabolism domain containing-1 (COMMD1), and phosphatase and tensin homolog (PTEN) were assayed using microscopic and biochemical methods The emphysematous areas were characterized by enlargement of the alveoli, destruction of the alveolar structure, accumulation of macrophages in the alveolar lumens, and showed increased HIF-1α immunoreactivity. Additionally, the emphysematous areas had significantly lower elastin, LOX, LOXL1, LOXL2, HIF-1α, COMMD1, and PTEN protein levels than the non-emphysematous areas. We suppose that the reductions in the HIF-1α levels led to decreases in the protein levels of active LOX, LOXL1, and LOXL2. These decreases might cause abnormalities in the elastic fiber biology. HIF-1α activation induced by decreased COMMD1 and protease activation induced by decreased PTEN might contribute to the development of PE. Finally, methods aimed at increasing the protein levels of LOXs, COMMD1 and PTEN might be effective for treating PE. Copyright © 2017 Elsevier GmbH. All rights reserved.

Protection of dopaminergic neurons by 5-lipoxygenase inhibitor.

PubMed

Kang, Kai-Hsiang; Liou, Horng-Hui; Hour, Mann-Jen; Liou, Houng-Chi; Fu, Wen-Mei

2013-10-01

Neuroinflammation and oxidative stress are important factors that induce neurodegeneration in age-related neurological disorders. 5-Lipoxygenase (5-LOX) is the enzyme responsible for catalysing the synthesis of leukotriene or 5-HETE from arachidonic acid. 5-LOX is expressed in the central nervous system and may cause neurodegenerative disease. In this study, we investigated the effect of the pharmacological inhibition of 5-lipoxygenase on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)/MPP(+)-induced dopaminergic neuronal death in midbrain neuron-glia co-cultures and in mice. It was found that 5-LOX was over-expressed in astrocytes after the injection of MPTP into C57BL6 mice. MK-886, a specific inhibitor of 5-LOX activating protein (FLAP), significantly increased [(3)H]-dopamine uptake, a functional indicator of the integrity of dopaminergic neurons, in midbrain cultures or the SH-SY5Y human dopaminergic cell line following MPP(+) treatment. In addition, LTB₄, one of 5-LOX's downstream products, was increased in the striatum and substantia nigra following MPTP injection in mice. LTB₄ but not LTD₄ and 5-HETE enhanced MPP(+)-induced neurotoxicity in primary midbrain cultures. MK-886 administration increased the number of tyrosine hydroxylase-positive neurons in the substantia nigra and the dopamine content in the striatum in MPTP-induced parkinsonian mice. Furthermore, the MPTP-induced upregulation of LTB₄ in the striatum and substantia nigra was antagonised by MK-886. These results suggest that 5-LOX inhibitors may be developed as novel neuroprotective agents and LTB₄ may play an important pathological role in Parkinson's disease. Copyright © 2013 Elsevier Ltd. All rights reserved.

Evaporation and combustion of LOX under supercritical and subcritical conditions

NASA Technical Reports Server (NTRS)

Yang, A. S.; Hsieh, W. H.; Kuo, K. K.

1993-01-01

The objective is to study the evaporation and combustion of LOX under supercritical and subcritical conditions both experimentally and theoretically. In the evaporation studies, evaporation rate and surface temperature were measured when LOX vaporizing in helium environments at pressures ranging from 5 to 68 atm. A Varian 3700 gas chromatograph was employed to measure the oxygen concentration above the LOX surface. For the combustion tests, high-magnification video photography was used to record direct images of the flame shape of a LOX/H2/He laminar diffusion flame. The gas composition in the post-flame region is also being measured with the gas sampling and chromatography analysis. These data are being used to validate the theoretical model. A comprehensive theoretical model with the consideration of the solubility of ambient gases as well as variable thermophysical properties was formulated and solved numerically to study the gasification and burning of LOX at elevated pressures. The calculated flame shape agreed reasonably well with the edge of the observed luminous flame surface. The effect of gravity on the flame structure of laminar diffusion flames was found to be significant. In addition, the predicted results using the flame-sheet model were compared with those based upon full equilibrium calculations (which considered the formation of intermediate species) at supercritical pressures. Except at the flame front where temperature exceeded 2,800 K, the flame-sheet and equilibrium solutions in terms of temperature distributions were in very close agreement. The temperature deviation in the neighborhood of the flame front is caused by the effect of high-temperature dissociation.

Lysyl oxidase expression is decreased in the developing diaphragm and lungs of nitrofen-induced congenital diaphragmatic hernia.

PubMed

Takahashi, Toshiaki; Friedmacher, Florian; Takahashi, Hiromizu; Daniel Hofmann, Alejandro; Puri, Prem

2015-02-01

Malformation of the nonmuscular tissue components in congenital diaphragmatic hernia (CDH) is thought to underlie the diaphragmatic defect, causing intrathoracic herniation of abdominal viscera and thus disturbing normal lung development. It has been shown that diaphragmatic and pulmonary morphogeneses require the structural integrity of connective tissue, and developmental mutations that inhibit the formation of extracellular matrix (ECM) result in CDH with hypoplastic lungs. Lysyl oxidase (lox), an extracellular enzyme that catalyzes the cross-linking of ECM proteins, plays an essential role during diaphragmatic and pulmonary development by controlling the formation of connective tissue. Furthermore, lox (-/-) knockouts exhibit abnormal connective tissue with diaphragmatic defects and impaired airway morphogenesis. We designed this study to investigate the hypothesis that diaphragmatic and pulmonary lox expression is decreased in the nitrofen-induced CDH model. Timed-pregnant Sprague-Dawley rats were exposed to either nitrofen or vehicle on gestational day 9 (D9), and fetuses were harvested on selected time points D15 and D18. The micro-dissected fetal diaphragms (n=48) and lungs (n=48) were divided into two groups: control and nitrofen-exposed samples (n=12 per specimen and time point, respectively). Diaphragmatic and pulmonary gene expression levels of lox were analyzed by quantitative real-time polymerase chain reaction. Immunohistochemical staining was performed to evaluate lox protein expression in diaphragms and lungs. Relative mRNA expression of lox was significantly reduced in diaphragms and lungs of nitrofen-exposed fetuses on D15 (0.29 ± 0.08 vs. 0.12 ± 0.05; p<0.05 and 0.52 ± 0.44 vs. 0.20 ± 0.04; p<0.05) and D18 (0.90 ± 0.25 vs. 0.57 ± 0.23; p<0.05 and 0.59 ± 0.26 vs. 0.35 ± 0.09; p<0.05) compared with controls. Diaphragmatic and pulmonary immunoreactivity of lox was markedly decreased in nitrofen-exposed fetuses on D15 and D18 compared with controls. Decreased lox expression during diaphragmatic development and lung branching morphogenesis may interfere with normal cross-linking of ECM proteins, disrupting the integrity of connective tissue, and contributing to the diaphragmatic defect and impaired airway formation in the nitrofen-induced CDH model. Georg Thieme Verlag KG Stuttgart · New York.

Nutrient loadings to streams of the continental United States from municipal and industrial effluent?

USGS Publications Warehouse

Maupin, Molly A.; Ivahnenko, Tamara

2011-01-01

Data from the United States Environmental Protection Agency Permit Compliance System national database were used to calculate annual total nitrogen (TN) and total phosphorus (TP) loads to surface waters from municipal and industrial facilities in six major regions of the United States for 1992, 1997, and 2002. Concentration and effluent flow data were examined for approximately 118,250 facilities in 45 states and the District of Columbia. Inconsistent and incomplete discharge locations, effluent flows, and effluent nutrient concentrations limited the use of these data for calculating nutrient loads. More concentrations were reported for major facilities, those discharging more than 1 million gallons per day, than for minor facilities, and more concentrations were reported for TP than for TN. Analytical methods to check and improve the quality of the Permit Compliance System data were used. Annual loads were calculated using "typical pollutant concentrations" to supplement missing concentrations based on the type and size of facilities. Annual nutrient loads for over 26,600 facilities were calculated for at least one of the three years. Sewage systems represented 74% of all TN loads and 58% of all TP loads. This work represents an initial set of data to develop a comprehensive and consistent national database of point-source nutrient loads. These loads can be used to inform a wide range of water-quality management, watershed modeling, and research efforts at multiple scales.

Airbreathing Hypersonic Vision-Operational-Vehicles Design Matrix

NASA Technical Reports Server (NTRS)

Hunt, James L.; Pegg, Robert J.; Petley, Dennis H.

1999-01-01

This paper presents the status of the airbreathing hypersonic airplane and space-access vision-operational-vehicle design matrix, with emphasis on horizontal takeoff and landing systems being studied at Langley; it reflects the synergies and issues, and indicates the thrust of the effort to resolve the design matrix including Mach 5 to 10 airplanes with global-reach potential, pop-up and dual-role transatmospheric vehicles and airbreathing launch systems. The convergence of several critical systems/technologies across the vehicle matrix is indicated. This is particularly true for the low speed propulsion system for large unassisted horizontal takeoff vehicles which favor turbines and/or perhaps pulse detonation engines that do not require LOX which imposes loading concerns and mission flexibility restraints.

Airbreathing Hypersonic Vision-Operational-Vehicles Design Matrix

NASA Technical Reports Server (NTRS)

Hunt, James L.; Pegg, Robert J.; Petley, Dennis H.

1999-01-01

This paper presents the status of the airbreathing hypersonic airplane and space-access vision-operational-vehicle design matrix, with emphasis on horizontal takeoff and landing systems being, studied at Langley, it reflects the synergies and issues, and indicates the thrust of the effort to resolve the design matrix including Mach 5 to 10 airplanes with global-reach potential, pop-up and dual-role transatmospheric vehicles and airbreathing launch systems. The convergence of several critical systems/technologies across the vehicle matrix is indicated. This is particularly true for the low speed propulsion system for large unassisted horizontal takeoff vehicles which favor turbines and/or perhaps pulse detonation engines that do not require LOX which imposes loading concerns and mission Flexibility restraints.

MC-1 LOX Pump Rotating Cavitation: Issue and Investigation

NASA Technical Reports Server (NTRS)

VanHooser, Katherine; Turner, J. (Technical Monitor)

2001-01-01

Leading edge tip deformation was noted on the LOX inducer from the component test unit at disassembly. The damage was in a location similar to a previous Fastrac water flow inducer failure. A team was formed to investigate the anomaly.

Self-acting geometry for noncontact seals

NASA Technical Reports Server (NTRS)

Allen, G. P.

1981-01-01

Performance ot two self acting seal designs for a liquid oxygen (LOX) turbopump was predicted over ranges of pressure differential and speed. Predictions were compared with test results. Performance of a radial face seal for LOX was predicted up to 448 N/cu cm and 147 m/sec. Performance of a segmented circumferential seal for helium was predicted up to 69 N/cu cm and 189 m/sec. Results confirmed predictions of noncontact operation. Qualitative agreement between test and analysis was found. The LOX face seal evidently operated with mostly liquid in the self acting geometry and mostly gas across the dam.

Liquid oxygen liquid acquisition device bubble point tests with high pressure lox at elevated temperatures

NASA Astrophysics Data System (ADS)

Jurns, J. M.; Hartwig, J. W.

2012-04-01

When transferring propellant in space, it is most efficient to transfer single phase liquid from a propellant tank to an engine. In earth's gravity field or under acceleration, propellant transfer is fairly simple. However, in low gravity, withdrawing single-phase fluid becomes a challenge. A variety of propellant management devices (PMDs) are used to ensure single-phase flow. One type of PMD, a liquid acquisition device (LAD) takes advantage of capillary flow and surface tension to acquire liquid. The present work reports on testing with liquid oxygen (LOX) at elevated pressures (and thus temperatures) (maximum pressure 1724 kPa and maximum temperature 122 K) as part of NASA's continuing cryogenic LAD development program. These tests evaluate LAD performance for LOX stored in higher pressure vessels that may be used in propellant systems using pressure fed engines. Test data shows a significant drop in LAD bubble point values at higher liquid temperatures, consistent with lower liquid surface tension at those temperatures. Test data also indicates that there are no first order effects of helium solubility in LOX on LAD bubble point prediction. Test results here extend the range of data for LOX fluid conditions, and provide insight into factors affecting predicting LAD bubble point pressures.

Changes in photosynthetic rate and stress volatile emissions through desiccation-rehydration cycles in desiccation-tolerant epiphytic filmy ferns (Hymenophyllaceae).

PubMed

Niinemets, Ülo; Bravo, León A; Copolovici, Lucian

2018-07-01

Exposure to recurrent desiccation cycles carries a risk of accumulation of reactive oxygen species that can impair leaf physiological activity upon rehydration, but changes in filmy fern stress status through desiccation and rewatering cycles have been poorly studied. We studied foliage photosynthetic rate and volatile marker compounds characterizing cell wall modifications (methanol) and stress development (lipoxygenase [LOX] pathway volatiles and methanol) through desiccation-rewatering cycles in lower-canopy species Hymenoglossum cruentum and Hymenophyllum caudiculatum, lower- to upper-canopy species Hymenophyllum plicatum and upper-canopy species Hymenophyllum dentatum sampled from a common environment and hypothesized that lower canopy species respond more strongly to desiccation and rewatering. In all species, rates of photosynthesis and LOX volatile emission decreased with progression of desiccation, but LOX emission decreased with a slower rate than photosynthesis. Rewatering first led to an emission burst of LOX volatiles followed by methanol, indicating that the oxidative burst was elicited in the symplast and further propagated to cell walls. Changes in LOX emissions were more pronounced in the upper-canopy species that had a greater photosynthetic activity and likely a greater rate of production of photooxidants. We conclude that rewatering induces the most severe stress in filmy ferns, especially in the upper canopy species. © 2018 John Wiley & Sons Ltd.

Effect of ω-3 and ω-9 fatty acid rich oils on lipoxygenases and cyclooxygenases enzymes and on the growth of a mammary adenocarcinoma model.

PubMed

Comba, Andrea; Maestri, Damian M; Berra, María A; Garcia, Carolina Paola; Das, Undurti N; Eynard, Aldo R; Pasqualini, María E

2010-10-08

Nutritional factors play a major role in cancer initiation and development. Dietary polyunsaturated fatty acids (PUFAs) have the ability to induce modifications in the activity of lipoxygenase (LOX) and cyclooxygenase (COX) enzymes that affect tumour growth. We studied the effect of two diets enriched in 6% Walnut and Peanut oils that are rich in ω-3 and ω9 PUFAs respectively on a murine mammary gland adenocarcinoma as compared with the control (C) that received commercial diet. Peanut oil enriched diet induced an increase in membrane arachidonic acid (AA) content and the cyclooxygenase enzyme derived 12-HHT (p < 0.05) and simultaneously showed decrease in 12-LOX, 15-LOX-2, 15-LOX-1 and PGE activities (p < 0.05) that corresponded to higher apoptosis and lower mitosis seen in this group (p < 0.05). Furthermore, Peanut oil group showed lower T-cell infiltration (p < 0.05), number of metastasis (p < 0.05) and tumour volume (p < 0.05) and longer survival rate compared to other groups. The results of the present study showed that Peanut oil-enriched diet protects against mammary cancer development by modulating tumour membrane fatty acids composition and LOX and COX enzyme activities.

Synthesis, cytotoxicity, cellular uptake and influence on eicosanoid metabolism of cobalt-alkyne modified fructoses in comparison to auranofin and the cytotoxic COX inhibitor Co-ASS.

PubMed

Ott, Ingo; Koch, Thao; Shorafa, Hashem; Bai, Zhenlin; Poeckel, Daniel; Steinhilber, Dieter; Gust, Ronald

2005-06-21

Propargylhexacarbonyldicobalt complexes with fructopyranose ligands were prepared and investigated for cytotoxicity in the MCF-7 human breast cancer cell line. The antiproliferative effects depended on the presence of isopropylidene protecting groups in the carbohydrate ligand and correlated with the cellular concentration of the complexes. IC(50) values of > 20 microM demonstrated that the fructose derivatives were only moderately active compared to the references auranofin and the aspirin (ASS) derivative [2-acetoxy(2-propynyl)benzoate]hexacarbonyldicobalt (Co-ASS). In continuation of our studies on the mode of action of cobalt-alkyne complexes we studied the influence of the compounds on the formation of 12-HHT (COX-1 product) and 12-HETE (12-LOX product) by human platelets as an indication of the interference in the eicosanoid metabolism, which is discussed as a target system of cytostatics. Co-ASS was an efficient COX-1 inhibitor without LOX inhibitory activity and auranofin inhibited both COX-1 and 12-LOX eicosanoid production. The missing activity of the fructopyranose complexes at the 12-LOX and the only moderate effects at COX-1 indicate that COX/LOX inhibition may be in part responsible for the pharmacological effects of auranofin and Co-ASS but not for those of the fructopyranose complexes.

LOX-mediated collagen crosslinking is responsible for fibrosis-enhanced metastasis

PubMed Central

Cox, Thomas R.; Bird, Demelza; Baker, Ann-Marie; Barker, Holly E.; Ho, Melisa W-Y.; Lang, Georgina; Erler, Janine T.

2013-01-01

Tumor metastasis is a highly complex, dynamic and inefficient process involving multiple steps, yet accounts for over 90% of cancer patient deaths. Although it has long been known that fibrotic signals enhance tumor progression and metastasis, the underlying molecular mechanisms are still unclear. Identifying events involved in creating environments that promote metastatic colonization and growth is critical for the development of effective cancer therapies. Here, we demonstrate a critical role for lysyl oxidase (LOX) in establishing a milieu within fibrosing tissues that is favorable to growth of metastastic tumor cells. We show that LOX-dependent collagen crosslinking is involved in creating a growth-permissive fibrotic microenvironment capable of supporting metastatic growth by enhancing tumor cell persistence and survival. We show that therapeutic targeting of LOX abrogates not only the extent to which fibrosis manifests, but also prevents fibrosis-enhanced metastatic colonization. Finally, we show that the LOX-mediated collagen cross-linking directly increases tumor cell proliferation, enhancing metastatic colonization and growth manifesting in vivo as increased metastasis. This is the first time that crosslinking of collagen I has been shown to enhance metastatic growth. These findings provide an important link between extracellular matrix homeostasis, fibrosis and cancer with important clinical implications for both the treatment of fibrotic disease and cancer. PMID:23345161

Liquid Oxygen Liquid Acquisition Device Bubble Point Tests with High Pressure LOX at Elevated Temperatures

NASA Technical Reports Server (NTRS)

Jurns, John M.; Hartwig, Jason W.

2011-01-01

When transferring propellant in space, it is most efficient to transfer single phase liquid from a propellant tank to an engine. In earth s gravity field or under acceleration, propellant transfer is fairly simple. However, in low gravity, withdrawing single-phase fluid becomes a challenge. A variety of propellant management devices (PMD) are used to ensure single-phase flow. One type of PMD, a liquid acquisition device (LAD) takes advantage of capillary flow and surface tension to acquire liquid. The present work reports on testing with liquid oxygen (LOX) at elevated pressures (and thus temperatures) (maximum pressure 1724 kPa and maximum temperature 122K) as part of NASA s continuing cryogenic LAD development program. These tests evaluate LAD performance for LOX stored in higher pressure vessels that may be used in propellant systems using pressure fed engines. Test data shows a significant drop in LAD bubble point values at higher liquid temperatures, consistent with lower liquid surface tension at those temperatures. Test data also indicates that there are no first order effects of helium solubility in LOX on LAD bubble point prediction. Test results here extend the range of data for LOX fluid conditions, and provide insight into factors affecting predicting LAD bubble point pressures.

Stability Analysis of Intertank Formed Skin/Stringer Compression Panel with Simulated Damage

NASA Technical Reports Server (NTRS)

Harper, David W.; Wingate, Robert J.

2012-01-01

The External Tank (ET) is a component of the Space Shuttle launch vehicle that contains fuel and oxidizer. During launch, the ET supplies the space shuttle main engines with liquid hydrogen and liquid oxygen. In addition to supplying fuel and oxidizer, it is the backbone structural component of the Space Shuttle. It is comprised of a liquid hydrogen (LH2) tank and a liquid oxygen (LOX) tank, which are separated by an Intertank. The Intertank is a stringer-stiffened cylindrical structure with hat-section stringers that are roll formed from aluminum-lithium alloy Al-2090. Cracks in the Intertank stringers of the STS-133 ET were noticed after a November 5, 2010 launch attempt. The cracks were approximately nine inches long and occurred on the forward end of the Intertank (near the LOX tank), along the fastener line, and were believed to have occurred while loading the ET with the cryogenic propellants. These cracks generated questions about the structural integrity of the Intertank. In order to determine the structural capability of the Intertank with varying degrees of damage, a finite element model (FEM) simulating a 1995 compression panel test was analyzed and correlated to test data. Varying degrees of damage were simulated in the FEM, and non-linear stability analyses were performed. The high degree of similarity between the compression panel and the Intertank provided confidence that the ET Intertank would have similar capabilities.

Effects of MK-886, a 5-lipoxygenase activating protein (FLAP) inhibitor, and 5-lipoxygenase deficiency on the forced swimming behavior of mice

PubMed Central

Uz, Tolga; Dimitrijevic, Nikola; Imbesi, Marta; Manev, Hari; Manev, Radmila

2008-01-01

A common biological pathway may contribute to the comorbidity of atherosclerosis and depression. Increased activity of the enzymatic 5-lipoxygenase (5-LOX; 5LO) pathway is a contributing factor in atherosclerosis and a 5-LOX inhibitor, MK-886, is beneficial in animal models of atherosclerosis. In the brain, MK-886 increases phosphorylation of the glutamate receptor subunit GluR1, and the increased phosphorylation of this receptor has been associated with antidepressant treatment. In this work, we evaluated the behavioral effects of MK-886 in an automated assay of mouse forced swimming, which identifies antidepressant activity as increased climbing behavior and/or decreased rest time. Whereas a single injection of MK-886 (3 and 10 mg/kg) did not affect forced swimming behaviors assayed 30 min later, 6 daily injections of 3 mg/kg MK-886 slightly increased climbing and significantly reduced rest time in wild-type mice but not in 5-LOX-deficient mice. A diet delivery of MK-886, 4 μg per 100 mg body-weight per day, required three weeks to affect forced swimming; it increased climbing behavior. Climbing behavior was also increased in naive 5-LOX-deficient mice compared to naive wild-type controls. These results suggest that 5-LOX inhibition and deficiency may be associated with antidepressant activity. Increased climbing in a forced swimming assay is a typical outcome of antidepressants that increase noradrenergic and dopaminergic activity. Interestingly, 5-LOX deficiency and MK-886 treatment have been shown to be capable of increasing the behavioral effects of a noradrenaline/dopamine-potentiating drug, cocaine. Future research is needed to evaluate the clinical relevance of our findings. PMID:18403121

12/15-Lipoxygenase Inhibition or Knockout Reduces Warfarin-Associated Hemorrhagic Transformation After Experimental Stroke.

PubMed

Liu, Yu; Zheng, Yi; Karatas, Hulya; Wang, Xiaoying; Foerch, Christian; Lo, Eng H; van Leyen, Klaus

2017-02-01

For stroke prevention, patients with atrial fibrillation typically receive oral anticoagulation. The commonly used anticoagulant warfarin increases the risk of hemorrhagic transformation (HT) when a stroke occurs; tissue-type plasminogen activator treatment is therefore restricted in these patients. This study was designed to test the hypothesis that 12/15-lipoxygenase (12/15-LOX) inhibition would reduce HT in warfarin-treated mice subjected to experimental stroke. Warfarin was dosed orally in drinking water, and international normalized ratio values were determined using a Coaguchek device. C57BL6J mice or 12/15-LOX knockout mice were subjected to transient middle cerebral artery occlusion with 3 hours severe ischemia (model A) or 2 hours ischemia and tissue-type plasminogen activator infusion (model B), with or without the 12/15-LOX inhibitor ML351. Hemoglobin was determined in brain homogenates, and hemorrhage areas on the brain surface and in brain sections were measured. 12/15-LOX expression was detected by immunohistochemistry. Warfarin treatment resulted in reproducible increased international normalized ratio values and significant HT in both models. 12/15-LOX knockout mice suffered less HT after severe ischemia, and ML351 reduced HT in wild-type mice. When normalized to infarct size, ML351 still independently reduced hemorrhage. HT after tissue-type plasminogen activator was similarly reduced by ML351. In addition to its benefits in infarct size reduction, 12/15-LOX inhibition also may independently reduce HT in warfarin-treated mice. ML351 should be further evaluated as stroke treatment in anticoagulated patients suffering a stroke, either alone or in conjunction with tissue-type plasminogen activator. © 2017 American Heart Association, Inc.

Low-energy N-ion beam biotechnology application in the induction of Thai jasmine rice mutant with improved seed storability

NASA Astrophysics Data System (ADS)

Semsang, Nuananong; Techarang, Jiranat; Yu, Liangdeng; Phanchaisri, Boonrak

2018-06-01

Low-energy heavy-ion beam is a novel biotechnology used for mutation induction in plants. We used a low-energy N-ion beam to induce mutations in Thai jasmine rice (Oryza sativa L. cv. KDML 105) to improve the yield and seed quality. Seeds of BKOS6, a Thai jasmine rice mutant previously induced by ion beams, were re-bombarded with 60-kV-accelerated N-ions (N++N2+) to fluences of 1-2 × 1016 ions/cm2. The resulting mutant, named HyKOS21, exhibited photoperiod insensitivity, semi-dwarfness, and high yield potential. Seed storability of the mutant was studied in natural and accelerated ageing conditions and compared to that of KDML 105 and six other Thai rice varieties. In both testing conditions, HyKOS21 mutant had the highest seed storability among the tested varieties. After storage in the natural condition for 18 months, HyKOS21 had a seed germination percentage nearly two times as that of the original KDML 105. Biochemical analysis showed that the lipid peroxidation level of the mutant seeds was the lowest among those of the tested varieties. Furthermore, an expression analysis of genes encoding lipoxygenase isoenzyme (lox1, lox2, and lox3) revealed that the mutant lacked expression of lox1 and lox2 and expressed only lox3 in seeds. These results may explain the improved seed longevity of the mutant after storage. This work provides further evidence of the modification of biological materials using a low-energy ion beam to produce rice mutants with improved yield and seed storability. The benefits of this technology, to create new varieties with improved values, could serve for local economic development.

Self-excising Cre/mutant lox marker recycling system for multiple gene integrations and consecutive gene deletions in Aspergillus oryzae.

PubMed

Zhang, Silai; Ban, Akihiko; Ebara, Naoki; Mizutani, Osamu; Tanaka, Mizuki; Shintani, Takahiro; Gomi, Katsuya

2017-04-01

In this study, we developed a self-excising Cre/loxP-mediated marker recycling system with mutated lox sequences to introduce a number of biosynthetic genes into Aspergillus oryzae. To construct the self-excising marker cassette, both the selectable marker, the Aspergillus nidulans adeA gene, and the Cre recombinase gene (cre), conditionally expressed by the xylanase-encoding gene promoter, were designed to be located between the mutant lox sequences, lox66 and lox71. However, construction of the plasmid failed, possibly owing to a slight expression of cre downstream of the fungal gene promoter in Escherichia coli. Hence, to avoid the excision of the cassette in E. coli, a 71-bp intron of the A. oryzae xynG2 gene was inserted into the cre gene. The A. oryzae adeA deletion mutant was transformed with the resulting plasmid in the presence of glucose, and the transformants were cultured in medium containing xylose as the sole carbon source. PCR analysis of genomic DNA from resultant colonies revealed the excision of both the marker and Cre expression construct, indicating that the self-excising marker cassette was efficient at removing the selectable marker. Using the marker recycling system, hyperproduction of kojic acid could be achieved in A. oryzae by the introduction of two genes that encode oxidoreductase and transporter. Furthermore, we also constructed an alternative marker recycling cassette bearing the A. nidulans pyrithiamine resistant gene (ptrA) as a dominant selectable marker. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Different expression profiles of the lysyl oxidases and matrix metalloproteinases in human ACL fibroblasts after co-culture with synovial cells.

PubMed

Wang, Chunli; Xu, Chunming; Chen, Rongfu; Yang, Li; Sung, Kl Paul

2018-02-12

Purposes The anterior cruciate ligament (ACL) has poor functional healing response. The synovial tissue surrounding ACL ligament might be a major regulator of the microenvironment in the joint cavity after ACL injury, thus affecting the repair process. Using transwell co-culture, this study explored the direct influence of human synovial cells (HSCs) on ACL fibroblasts (ACLfs) by characterizing the differential expression of the lysyl oxidase family (LOXs) and matrix metalloproteinases (MMP-1, -2, -3), which facilitate extracellular matrix (ECM) repair and degradation, respectively. Methods The mRNA expression levels of LOXs and MMP-1, -2, -3 were analyzed by semi-quantitative PCR and quantitative real-time PCR. The protein expression levels of LOXs and MMP-1, -2, -3 were detected by western blot. Results We found that co-culture resulted in an increase in the mRNAs of LOXs in normal ACLfs and differentially regulated the expression of MMPs. Then we applied 12% mechanical stretch on ACLfs to induce injury and found the mRNA expression levels of LOXs in injured ACLfs were decreased in the co-culture group relative to the mono-culture group. Conversely, the mRNA expression levels of MMPs in injured ACLfs were promoted in the co-culture group compared with the mono-culture group. At translational level, we found that LOXs were lower while MMPs were highly expressed in the co-culture group compared to the mono-culture group. Conclusions The co-culture of ACLfs and HSCs, which mimicked the cell-to-cell contact in a micro-environment, could contribute to protein modulators for wound healing, inferring the potential reason for the poor self-healing of injured ACL.

The Oxidized Low-Density Lipoprotein Receptor Mediates Vascular Effects of Inhaled Vehicle Emissions

PubMed Central

Lucero, JoAnn; Harman, Melissa; Madden, Michael C.; McDonald, Jacob D.; Seagrave, Jean Clare; Campen, Matthew J.

2011-01-01

Rationale: To determine vascular signaling pathways involved in inhaled air pollution (vehicular engine emission) exposure–induced exacerbation of atherosclerosis that are associated with onset of clinical cardiovascular events. Objectives: To elucidate the role of oxidized low-density lipoprotein (oxLDL) and its primary receptor on endothelial cells, the lectin-like oxLDL receptor (LOX-1), in regulation of endothelin-1 expression and matrix metalloproteinase activity associated with inhalational exposure to vehicular engine emissions. Methods: Atherosclerotic apolipoprotein E knockout mice were exposed by inhalation to filtered air or mixed whole engine emissions (250 μg particulate matter [PM]/m3 diesel + 50 μg PM/m3 gasoline exhausts) 6 h/d for 7 days. Concurrently, mice were treated with either mouse IgG or neutralizing antibodies to LOX-1 every other day. Vascular and plasma markers of oxidative stress and expression proatherogenic factors were assessed. In a parallel study, healthy human subjects were exposed to either 100 μg PM/m3 diesel whole exhaust or high-efficiency particulate air and charcoal-filtered “clean” air (control subjects) for 2 hours, on separate occasions. Measurements and Main Results: Mixed emissions exposure increased oxLDL and vascular reactive oxygen species, as well as LOX-1, matrix metalloproteinase-9, and endothelin-1 mRNA expression and also monocyte/macrophage infiltration, each of which was attenuated with LOX-1 antibody treatment. In a parallel study, diesel exhaust exposure in volunteer human subjects induced significant increases in plasma-soluble LOX-1. Conclusions: These findings demonstrate that acute exposure to vehicular source pollutants results in up-regulation of vascular factors associated with progression of atherosclerosis, endothelin-1, and matrix metalloproteinase-9, mediated through oxLDL–LOX-1 receptor signaling, which may serve as a novel target for future therapy. PMID:21493736

MiR-106a Associated with Diabetic Peripheral Neuropathy Through the Regulation of 12/15-LOX-meidiated Oxidative/Nitrative Stress.

PubMed

Wu, Yuanbo; Xu, Dandan; Zhu, Xiang; Yang, Guangwei; Ren, Mingshan

2017-01-01

Diabetic peripheral neuropathy (DPN) is a common complication of diabetes with a complex pathogenesis. Study has reported that oxidative stress and nitrative stress are all involved in the DPN. However, the mechanism between them is still unclear. In the present study, we investigated whether miR-106a regulated 12/15-Lipoxygenase (12/15-LOX) of oxidative/nitrative stress (OS/NS) in DPN. Dorsal root ganglion (DRG) was isolated from 10 healthy mice and 10 DPN mice. DPN mouse model was established by the injection of streptozotocin (STZ), and high glucose was used for inducing the in vitro model of DPN. In DPN mice, the levels of fasting blood-glucose (FBG) level, Methane Dicarboxylic Aldehyde (MDA) and the Inducible Nitric Oxide Synthase (iNOS) were increased, while the levels of motor nerve conduction velocity (MNCV), sensory nerve conduction velocity (SNCV) and superoxide dismutase (SOD) were decreased. MiR-106a level in DRG cells of DPN was decreased. The 12/15-LOX expression and cell apoptosis were increased. DRG cells subjected to high glucose resulted in the same effects as above. MiR-106a was observed to target 12/15-LOX and regulated its expression. MiR-106a overexpression reversed the effect that was induced by high glucose, however, overexpression of 12/15-LOX reversed the effect that was induced by miR-106a overexpression. MiR-106 may be associated with 12/15-LOX mediated OS/NS in DPN and may be considered as a potential therapeutic target. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Effects of elevated [CO2] on maize defence against mycotoxigenic Fusarium verticillioides

PubMed Central

Vaughan, Martha M; Huffaker, Alisa; Schmelz, Eric A; Dafoe, Nicole J; Christensen, Shawn; Sims, James; Martins, Vitor F; Swerbilow, Jay; Romero, Maritza; Alborn, Hans T; Allen, Leon HARTWELL; Teal, Peter EA

2014-01-01

Maize is by quantity the most important C4 cereal crop; however, future climate changes are expected to increase maize susceptibility to mycotoxigenic fungal pathogens and reduce productivity. While rising atmospheric [CO2] is a driving force behind the warmer temperatures and drought, which aggravate fungal disease and mycotoxin accumulation, our understanding of how elevated [CO2] will effect maize defences against such pathogens is limited. Here we report that elevated [CO2] increases maize susceptibility to Fusarium verticillioides proliferation, while mycotoxin levels are unaltered. Fumonisin production is not proportional to the increase in F. verticillioides biomass, and the amount of fumonisin produced per unit pathogen is reduced at elevated [CO2]. Following F. verticillioides stalk inoculation, the accumulation of sugars, free fatty acids, lipoxygenase (LOX) transcripts, phytohormones and downstream phytoalexins is dampened in maize grown at elevated [CO2]. The attenuation of maize 13-LOXs and jasmonic acid production correlates with reduced terpenoid phytoalexins and increased susceptibility. Furthermore, the attenuated induction of 9-LOXs, which have been suggested to stimulate mycotoxin biosynthesis, is consistent with reduced fumonisin per unit fungal biomass at elevated [CO2]. Our findings suggest that elevated [CO2] will compromise maize LOX-dependent signalling, which will influence the interactions between maize and mycotoxigenic fungi. Elevated [CO2] increases maize susceptibility to Fusarium verticillioides proliferation but mycotoxin levels are unaltered. The attenuation of maize 13-LOXs and JA production correlates with reduced terpenoid phytoalexins and increased susceptibility. Furthermore, the attenuated induction of 9-LOXs, which have been suggested to stimulate mycotoxin biosynthesis, is consistent with reduced fumonisin per unit fungal biomass at elevated [CO2]. PMID:24689748

Structure Based Library Design (SBLD) for new 1,4-dihydropyrimidine scaffold as simultaneous COX-1/COX-2 and 5-LOX inhibitors.

PubMed

Lokwani, Deepak; Azad, Rajaram; Sarkate, Aniket; Reddanna, Pallu; Shinde, Devanand

2015-08-01

The various scaffolds containing 1,4-dihydropyrimidine ring were designed by considering the environment of the active site of COX-1/COX-2 and 5-LOX enzymes. The structure-based library design approach, including the focused library design (Virtual Combinatorial Library Design) and virtual screening was used to select the 1,4-dihydropyrimidine scaffold for simultaneous inhibition of both enzyme pathways (COX-1/COX-2 and 5-LOX). The virtual library on each 1,4-dihydropyrimidine scaffold was enumerated in two alternative ways. In first way, the chemical reagents at R groups were filtered by docking of scaffold with single position substitution, that is, only at R1, or R2, or R3, … Rn on COX-2 enzyme using Glide XP docking mode. The structures that do not dock well were removed and the library was enumerated with filtered chemical reagents. In second alternative way, the single position docking stage was bypassed, and the entire library was enumerated using all chemical reagents by docking on the COX-2 enzyme. The entire library of approximately 15,629 compounds obtained from both ways after screening for drug like properties, were further screened for their binding affinity against COX-1 and 5-LOX enzymes using Virtual Screening Workflow. Finally, 142 hits were obtained and divided into two groups based on their binding affinity for COX-1/COX-2 and for both enzyme pathways (COX-1/COX-2 and 5-LOX). The ten molecules were selected, synthesized and evaluated for their COX-1, COX-2 and 5-LOX inhibiting activity. Copyright © 2015 Elsevier Ltd. All rights reserved.

Rocketdyne LOX bearing tester program

NASA Technical Reports Server (NTRS)

Keba, J. E.; Beatty, R. F.

1988-01-01

The cause, or causes, for the Space Shuttle Main Engine ball wear were unknown, however, several mechanisms were suspected. Two testers were designed and built for operation in liquid oxygen to empirically gain insight into the problems and iterate solutions in a timely and cost efficient manner independent of engine testing. Schedules and test plans were developed that defined a test matrix consisting of parametric variations of loading, cooling or vapor margin, cage lubrication, material, and geometry studies. Initial test results indicated that the low pressure pump thrust bearing surface distress is a function of high axial load. Initial high pressure turbopump bearing tests produced the wear phenomenon observed in the turbopump and identified an inadequate vapor margin problem and a coolant flowrate sensitivity issue. These tests provided calibration data of analytical model predictions to give high confidence in the positive impact of future turbopump design modification for flight. Various modifications will be evaluated in these testers, since similar turbopump conditions can be produced and the benefit of the modification will be quantified in measured wear life comparisons.

Wear of liquid nitrogen-cooled 440C bearing steels in an oxygen environment

NASA Technical Reports Server (NTRS)

Chaudhuri, Dilip K.; Verma, Ravi

1988-01-01

This paper presents up-to-date findings of the research being conducted to understand the mechanism of sliding wear in unlubricated 440C bearing steels under oxidative conditions. A sliding wear test rig has been designed and built with a cylinder-on-flat geometry. The equipment is capable of testing specimens under high axial loads and sliding speeds in a simulated LOX environment. Samples of 440C steel, quenched and tempered to a hardness of Rc 56, were tested under a load of 890 N and a sliding speed of 2.05 m/sec for total sliding distances of up to 5.54 km. Flash temperatures during these tests were measured with an IR camera and a fast digital recorder. Microstructural and microanalytical data from the worn surfaces and the debris particles are analyzed extensively, along with wear rates, flash temperatures, surface profiles, hardnesses, and residual stresses, in the context of oxidation and wear theories.

Comparative anti-psoriatic efficacy studies of clobetasol loaded chitin nanogel and marketed cream.

PubMed

Panonnummal, Rajitha; Jayakumar, R; Sabitha, M

2017-01-01

In the present study chitin nanogel loaded with anti-psoriatic drug clobetasol was developed (CLCNG) for its topical delivery in psoriasis. CLCNG had the particle size of 132±14nm, with gel like consistency, stability in refrigerator, having higher drug release properties at acidic pH. CLCNG exhibited significant toxicity towards HaCaT and THP-1cell lines by MTT assay. The uptake of nanogel by HaCaT cell lines was confirmed by fluorescent microscopy. CLCNG at 0.35mg/ml exhibited significant anti-inflammatory activity with an average of 65% and 70% inhibition in COX and LOX activities expressed in THP-1 cells. In vitro skin permeation studies revealed the increased transdermal flux with fragmented stratum corneum and loosened epidermal layers in CLCNG treated samples, compared with control drug solution. The in vivo anti-psoriatic studies done on imiquimod model confirmed the potential benefits of the nanogel for the topical delivery of clobetasol in psoriasis. Copyright © 2016 Elsevier B.V. All rights reserved.

A role for 9-lipoxygenases in maize defense against insect herbivory.

PubMed

Woldemariam, Melkamu G; Ahern, Kevin; Jander, Georg; Tzin, Vered

2018-01-02

Feeding by Spodoptera exigua (beet armyworm) larvae on Zea mays (maize) induces expression of 9-lipoxygenases to a greater extent than 13-lipoxygenases. Whereas 13-lipoxygenases have an established role in the synthesis of jasmonates that serve as defense signaling molecules in many plant species, relatively little is known about the role of 9-lipoxygenases in herbivore defense. Phylogenetic analysis of lipoxygenases from maize inbred lines B73 and W22 shows that, although most Lox genes are present in both lines, Lox12, a 9-lipoxygenase that has been implicated in fungal defense, is truncated and unlikely to encode a functional protein in W22. Two independent Mutator transposon insertions in another 9-lipoxygenase, Lox4, caused improved S. exigua growth on the mutant lines relative to wildtype W22. This observation suggests a function in herbivore defense for metabolic products downstream of maize Lox4, either through direct toxicity or a perhaps an as yet unknown signaling function.

Control of lens development by Lhx2-regulated neuroretinal FGFs

PubMed Central

Thein, Thuzar; de Melo, Jimmy; Zibetti, Cristina; Clark, Brian S.; Juarez, Felicia

2016-01-01

Fibroblast growth factor (FGF) signaling is an essential regulator of lens epithelial cell proliferation and survival, as well as lens fiber cell differentiation. However, the identities of these FGF factors, their source tissue and the genes that regulate their synthesis are unknown. We have found that Chx10-Cre;Lhx2lox/lox mice, which selectively lack Lhx2 expression in neuroretina from E10.5, showed an early arrest in lens fiber development along with severe microphthalmia. These mutant animals showed reduced expression of multiple neuroretina-expressed FGFs and canonical FGF-regulated genes in neuroretina. When FGF expression was genetically restored in Lhx2-deficient neuroretina of Chx10-Cre;Lhx2lox/lox mice, we observed a partial but nonetheless substantial rescue of the defects in lens cell proliferation, survival and fiber differentiation. These data demonstrate that neuroretinal expression of Lhx2 and neuroretina-derived FGF factors are crucial for lens fiber development in vivo. PMID:27633990

Synthesis and Structure–Activity Relationship Studies of 4-((2-Hydroxy-3-methoxybenzyl)amino)benzenesulfonamide Derivatives as Potent and Selective Inhibitors of 12-Lipoxygenase

PubMed Central

Luci, Diane K.; Jameson, J. Brian; Yasgar, Adam; Diaz, Giovanni; Joshi, Netra; Kantz, Auric; Markham, Kate; Perry, Steve; Kuhn, Norine; Yeung, Jennifer; Kerns, Edward H.; Schultz, Lena; Holinstat, Michael; Nadler, Jerry L.; Taylor-Fishwick, David A.; Jadhav, Ajit; Simeonov, Anton; Holman, Theodore R.; Maloney, David J.

2014-01-01

Human lipoxygenases (LOXs) are a family of iron-containing enzymes which catalyze the oxidation of polyunsaturated fatty acids to provide the corresponding bioactive hydroxyeicosatetraenoic acid (HETE) metabolites. These eicosanoid signaling molecules are involved in a number of physiologic responses such as platelet aggregation, inflammation, and cell proliferation. Our group has taken a particular interest in platelet-type 12-(S)-LOX (12-LOX) because of its demonstrated role in skin diseases, diabetes, platelet hemostasis, thrombosis, and cancer. Herein, we report the identification and medicinal chemistry optimization of a 4-((2-hydroxy-3-methoxybenzyl)amino)benzenesulfonamide-based scaffold. Top compounds, exemplified by 35 and 36, display nM potency against 12-LOX, excellent selectivity over related lipoxygenases and cyclooxygenases, and possess favorable ADME properties. In addition, both compounds inhibit PAR-4 induced aggregation and calcium mobilization in human platelets and reduce 12-HETE in β-cells. PMID:24393039

Fundamental Boiling and RP-1 Freezing Experiments

NASA Technical Reports Server (NTRS)

Goode, Brian

2002-01-01

The prestart thermal conditioning of the hardware in LOX (liquid oxygen) systems involve heat transfer between LOX and metal where boiling plays a large role. Information is easily found on nucleate boiling, maximum heat flux, minimum heat flux and film boiling for common fluids like water. After looking at these standard correlations it was felt more data was needed for the cool down side transition boiling for the LN2 and LOX. In particular interest is the film boiling values, the temperature at which transition begins and the slope as peak heat flux is approached. The ultimate goal is an array of boiling heat transfer coefficient as a function of surface temperature which can be used in the chilldown model of the feed system, engine and bleed system for X-34. The first experiment consisted of an actual MC-1 LOX Impeller which had been machined backwards, that was instrumented with 17 surface thermocouples and submerged in liquid nitrogen. The thermocouples were installed on metal thicknesses varying from the thin inducer to the thick hub.

Design, development and calibration of HTS wire based LOX level sensor probe

NASA Astrophysics Data System (ADS)

Karunanithi, R.; Jacob, S.; Nadig, D. S.; Prasad, M. V. N.; Gour, A. S.; Gowthaman, M.; Deekshith, P.; Shrivastava, V.

2014-01-01

For space applications, the weight of the liquid level sensors are of major concern as they affect the payload fraction and hence the cost. An attempt is made to design and test a light weight High Temperature Superconductor (HTS) wire based liquid level sensor for Liquid Oxygen (LOX) tank used in the cryostage of the spacecraft. The total resistance value measured of the HTS wire is inversely proportional to the liquid level. A HTS wire (SF12100) of 12mm width and 2.76m length without copper stabilizer has been used in the level sensor. The developed HTS wire based LOX level sensor is calibrated against a discrete diode array type level sensor. Liquid Nitrogen (LN2) and LOX has been used as cryogenic fluid for the calibration purpose. The automatic data logging for the system has been done using LabVIEW11. The net weight of the developed sensor is less than 1 kg.

Combustion performance and heat transfer characterization of LOX/hydrocarbon type propellants

NASA Technical Reports Server (NTRS)

Michel, R. W.

1983-01-01

An evaluation liquid oxygen (LOX) and various hydrocarbon fuels as low cost alternative propellants suitable for future space transportation system applications was done. The emphasis was directed toward low earth orbit maneuvering engine and reaction control engine systems. The feasibility of regeneratively cooling an orbit maneuvering thruster was analytically determined over a range of operating conditions from 100 to 1000 psia chamber pressure and 1000 to 10,000-1bF thrust, and specific design points were analyzed in detail for propane, methane, RP-1, ammonia, and ethanol; similar design point studies were performed for a film-cooled reaction control thruster. Heat transfer characteristics of propane were experimentally evaluated in heated tube tests. Forced convection heat transfer coefficients were determined. Seventy-seven hot firing tests were conducted with LOX/propane and LOX/ethanol, for a total duration of nearly 1400 seconds, using both heat sink and water-cooled calorimetric chambers. Combustion performance and stability and gas-side heat transfer characteristics were evaluated.

The protective effect of bergamot oil extract on lecitine-like oxyLDL receptor-1 expression in balloon injury-related neointima formation.

PubMed

Mollace, Vincenzo; Ragusa, Salvatore; Sacco, Iolanda; Muscoli, Carolina; Sculco, Francesca; Visalli, Valeria; Palma, Ernesto; Muscoli, Saverio; Mondello, Luigi; Dugo, Paola; Rotiroti, Domenicantonio; Romeo, Francesco

2008-06-01

Lectin-like oxyLDL receptor-1 (LOX-1) has recently been suggested to be involved in smooth muscle cell (SMC) proliferation and neointima formation in injured blood vessels. This study evaluates the effect of the nonvolatile fraction (NVF), the antioxidant component of bergamot essential oil (BEO), on LOX-1 expression and free radical generation in a model of rat angioplasty. Common carotid arteries injured by balloon angioplasty were removed after 14 days for histopathological, biochemical, and immunohistochemical studies. Balloon injury led to a significant restenosis with SMC proliferation and neointima formation, accompanied by increased expression of LOX-1 receptor, malondialdehyde and superoxide formation, and nitrotyrosine staining. Pretreatment of rats with BEO-NVF reduced the neointima proliferation together with free radical formation and LOX-1 expression in a dose-dependent manner. These results suggest that natural antioxidants may be relevant in the treatment of vascular disorders in which proliferation of SMCs and oxyLDL-related endothelial cell dysfunction are involved.

Physalis alkekengi Carotenoidic Extract Inhibitor of Soybean Lipoxygenase-1 Activity

PubMed Central

Chedea, Veronica Sanda; Pintea, Adela; Bunea, Andrea; Braicu, Cornelia; Stanila, Andreea; Socaciu, Carmen

2014-01-01

The aim of this study was to evaluate the effect of the carotenoidic saponified extract of Physalis alkekengi sepals (PA) towards the lipoxygenase (LOX) oxidation of linoleic acid. Lipoxygenase activity in the presence of carotenoids, standard and from extract, was followed by its kinetic behaviour determining the changes in absorption at 234 nm. The standard carotenoids used were β-carotene (β-car), lutein (Lut), and zeaxanthin (Zea). The calculated enzymatic specific activity (ESA) after 600 s of reaction proves that PA carotenoidic extract has inhibitory effect on LOX oxidation of linoleic acid. A longer polyenic chain of carotenoid structure gives a higher ESA during the first reaction seconds. This situation is not available after 600 s of reaction and may be due to a destruction of this structure by cooxidation of carotenoids, besides the classical LOX reaction. The PA carotenoidic extract inhibiting the LOX-1 reaction can be considered a source of lipoxygenase inhibitors. PMID:24511537

Identification of Protein Targets of 12/15-Lipoxygenase-Derived Lipid Electrophiles in Mouse Peritoneal Macrophages Using Omega-Alkynyl Fatty Acid.

PubMed

Isobe, Yosuke; Kawashima, Yusuke; Ishihara, Tomoaki; Watanabe, Kenji; Ohara, Osamu; Arita, Makoto

2018-04-20

The 12/15-lipoxygenase (12/15-LOX) enzyme introduces peroxyl groups, in a position-specific manner, into polyunsaturated fatty acids to form various kinds of bioactive lipid metabolites, including lipid-derived electrophiles (LDE). The resident peritoneal macrophage is the site of highest 12/15-LOX expression in the mouse. However, the role of the enzyme in the regulation of resident macrophages is not fully understood. Here, we describe a chemoproteomic method to identify the targets of enzymatically generated LDE. By treating mouse peritoneal macrophages with omega-alkynyl arachidonic acid (aAA), we identified a series of proteins adducted by LDE generated through a 12/15-LOX catalyzed reaction. Pathway analysis revealed a dramatic enrichment of proteins involved in energy metabolism and found that glycolytic flux and mitochondrial respiration were significantly affected by the expression of 12/15-LOX. Our findings thus highlight the utility of chemoproteomics using aAA for identifying intracellular targets of enzymatically generated LDE.

Preparation for Testing a Multi-Bay Box Subjected to Combined Loads

NASA Technical Reports Server (NTRS)

Rouse, Marshall; Jegley, Dawn

2015-01-01

The COmbined Loads Test System (COLTS) facility at NASA Langley Research Center provides a test capability to help develop validated structures technologies. The test machine was design to accommodate a range of fuselage structures and wing sections and subject them to both quasistatic and cyclic loading conditions. The COLTS facility is capable of testing fuselage barrels up to 4.6 m in diameter and 13.7 m long with combined mechanical, internal pressure, and thermal loads. The COLTS facility is currently being prepared to conduct a combined mechanical and pressure loading for a multi-bay pressure box to experimentally verify the structural performance of a composite structure which is 9.1 meters long and representative of a section of a hybrid wing body fuselage section in support of the Environmentally Responsible Aviation Project at NASA. This paper describes development of the multi-bay pressure box test using the COLTS facility. The multi-bay test article will be subjected to mechanical loads and internal pressure loads up to design ultimate load. Mechanical and pressure loads will be applied independently in some tests and simultaneously in others.

Space Shuttle Main Engine structural analysis and data reduction/evaluation. Volume 5: Main Injector LOX Inlet analysis

NASA Technical Reports Server (NTRS)

Violett, Rebeca S.

1989-01-01

The analysis performed on the Main Injector LOX Inlet Assembly located on the Space Shuttle Main Engine is summarized. An ANSYS finite element model of the inlet assemably was built and executed. Static stress analysis was also performed.

40 CFR 63.11088 - What requirements must I meet for gasoline loading racks if my facility is a bulk gasoline...

Code of Federal Regulations, 2014 CFR

2014-07-01

... gasoline loading racks if my facility is a bulk gasoline terminal, pipeline breakout station, or pipeline... CATEGORIES (CONTINUED) National Emission Standards for Hazardous Air Pollutants for Source Category: Gasoline... § 63.11088 What requirements must I meet for gasoline loading racks if my facility is a bulk gasoline...

40 CFR 63.11088 - What requirements must I meet for gasoline loading racks if my facility is a bulk gasoline...

Code of Federal Regulations, 2010 CFR

2010-07-01

... gasoline loading racks if my facility is a bulk gasoline terminal, pipeline breakout station, or pipeline... CATEGORIES (CONTINUED) National Emission Standards for Hazardous Air Pollutants for Source Category: Gasoline... § 63.11088 What requirements must I meet for gasoline loading racks if my facility is a bulk gasoline...

40 CFR 63.11088 - What requirements must I meet for gasoline loading racks if my facility is a bulk gasoline...

Code of Federal Regulations, 2013 CFR

2013-07-01

... gasoline loading racks if my facility is a bulk gasoline terminal, pipeline breakout station, or pipeline... CATEGORIES (CONTINUED) National Emission Standards for Hazardous Air Pollutants for Source Category: Gasoline... § 63.11088 What requirements must I meet for gasoline loading racks if my facility is a bulk gasoline...

40 CFR 63.11088 - What requirements must I meet for gasoline loading racks if my facility is a bulk gasoline...

Code of Federal Regulations, 2011 CFR

2011-07-01

... gasoline loading racks if my facility is a bulk gasoline terminal, pipeline breakout station, or pipeline... CATEGORIES (CONTINUED) National Emission Standards for Hazardous Air Pollutants for Source Category: Gasoline... § 63.11088 What requirements must I meet for gasoline loading racks if my facility is a bulk gasoline...

40 CFR 63.11088 - What requirements must I meet for gasoline loading racks if my facility is a bulk gasoline...

Code of Federal Regulations, 2012 CFR

2012-07-01

... gasoline loading racks if my facility is a bulk gasoline terminal, pipeline breakout station, or pipeline... CATEGORIES (CONTINUED) National Emission Standards for Hazardous Air Pollutants for Source Category: Gasoline... § 63.11088 What requirements must I meet for gasoline loading racks if my facility is a bulk gasoline...

5-Lipoxygenase Inhibition of the Fructus of Foeniculum vulgare and Its Constituents

PubMed Central

Lee, Je Hyeong; Lee, Dong Ung; Kim, Yeong Shik; Kim, Hyun Pyo

2012-01-01

The fruits of Foeniculum vulgare (Foeniculi Fructus) have been widely used in Chinese medicine as an antiemetic, ameliorating stomach ailments and as an analgesic. In order to establish its potential for antiallergic use, inhibitory actions of the fruiton 5-lipoxgenase (5-LOX) and β-hexosaminidase release were evaluated. The 70% ethanol extract of this plant material (FR) considerably inhibited 5-LOX-catalyzed leukotriene production from A23187-induced rat basophilic leukemia (RBL)-1 cells. The IC50 was 3.2 μg/ml. From this extract, 12 major compounds including sabinene, fenchone, γ-terpinene, α-pinene, limonene, p-anisylacetone, p-anisylaldehyde, estragole (4-allylanisole), trans-anethole, scopoletin, bergapten and umbelliferone were isolated. And it was found that several terpene derivatives including γ-terpinene and fenchone as well as phenylpropanoid, trans-anethole, showed considerable inhibitory action of 5-LOX. In particular, the IC50 of trans-anethole was 51.6 μ M. In contrast, FR and the isolated compounds did not show considerable inhibitory activity on the degranulation reaction of β-hexosaminidase release from antigen-treated RBL-2H3 cells. Against arachidonic acid-induced ear edema in mice, FR and trans-anethole showed significant inhibition by oral administration at doses of 100-400 mg/kg. In conclusion, FR and several major constituents are 5-LOX inhibitors and they may have potential for treating 5-LOX-related disorders. PMID:24116283

Anatomy of the lobula complex in the brain of the praying mantis compared to the lobula complexes of the locust and cockroach

PubMed Central

von Hadeln, Joss; Salden, Tobias; Homberg, Uwe

2017-01-01

Abstract The praying mantis is an insect which relies on vision for capturing prey, avoiding being eaten and for spatial orientation. It is well known for its ability to use stereopsis for estimating the distance of objects. The neuronal substrate mediating visually driven behaviors, however, is not very well investigated. To provide a basis for future functional studies, we analyzed the anatomical organization of visual neuropils in the brain of the praying mantis Hierodula membranacea and provide supporting evidence from a second species, Rhombodera basalis, with particular focus on the lobula complex (LOX). Neuropils were three‐dimensionally reconstructed from synapsin‐immunostained whole mount brains. The neuropil organization and the pattern of γ‐aminobutyric acid immunostaining of the medulla and LOX were compared between the praying mantis and two related polyneopteran species, the Madeira cockroach and the desert locust. The investigated visual neuropils of the praying mantis are highly structured. Unlike in most insects the LOX of the praying mantis consists of five nested neuropils with at least one neuropil not present in the cockroach or locust. Overall, the mantis LOX is more similar to the LOX of the locust than the more closely related cockroach suggesting that the sensory ecology plays a stronger role than the phylogenetic distance of the three species in structuring this center of visual information processing. PMID:28295329

Seal Technology for Liquid Oxygen (LOX) Turbopumps

NASA Technical Reports Server (NTRS)

Shapiro, Wilbur; Hamm, Robert

1985-01-01

Two types of advanced seals for liquid oxygen (LOX) turbopumps were investigated. One was a spiral-groove face seal whose function is to seal high-pressure LOX at the impeller end of the turbopump. The other was a floating-ring, Rayleigh-step, helium buffered seal used to prevent LOX ingress to the turbine side of the unit. For each seal type, two sizes were investigated (50 and 20 mm). A turbine-driven test rig was designed and manufactured, and a test program was completed on the 50 mm floating-ring, Rayleigh-step, helium buffered seal. Significant results were: vaporization in the flow path could cause failure by overheating; therefore, the spiral-groove pumping portion of the seal that provides the fluid film must circulate fluid without disruption if vaporization occurs in the sealing dam. This is successfully accomplished by a pressure-balanced spiral-groove concept that is described. The spiral-groove configuration is affected by turbulence in the fluid film and pressure drops due to fluid inertia at sudden contractions. The net results of these effects are deep grooves, large operating films, and high power loss when compared against seals operating with laminar films. Turbulence and inertia are induced by the high-density and low-viscosity characteristics of LOX. The program clearly pointed out the need to consider system environmental factors such as thermal and centrifugal distortions and rotor vibrations in the seal design.

CJ-13610, an orally active inhibitor of 5-lipoxygenase is efficacious in preclinical models of pain.

PubMed

Cortes-Burgos, Luz A; Zweifel, Ben S; Settle, Steven L; Pufahl, Robert A; Anderson, Gary D; Hardy, Medora M; Weir, Dana E; Hu, George; Happa, Fernando A; Stewart, Zachary; Muthian, Shanmugam; Graneto, Matthew J; Masferrer, Jaime L

2009-09-01

Zileuton, a redox and iron chelator 5-lipoxygenase (5-LOX) inhibitor and, leukotriene receptor antagonists are presently used clinically in the long term treatment of asthma. Recent data implicate 5-LOX pathway in pain signaling. We report 5-LOX expression in the central nervous system (CNS) and analyze the pain efficacy of a new class of non redox, non iron chelating 5-LOX inhibitor. CJ-13610, 4-(3-(4-(2-methyl-1H-imidazol-1-yl) phenylthio) phenyl)-tetrahydro-2H-pyran-4-carboxamide, demonstrated antihyperalgesic activity in inflammatory pain models including the acute carrageenan model and the chronic inflammatory model using complete Freund's adjuvant. Following complete Freund's adjuvant stimulus leukotrieneB(4) concentration in the brain was elevated (9+/-1 ng/g, mean+/-S.E.M.) by about 3 times that of the control group (3+/-0.11, mean+/-S.E.M.). Hyperalgesia and leukotrieneB(4) concentration were both reversed following CJ-13610 treatment. Furthermore, we demonstrate CJ-13610 efficacy against osteoarthritis like pain using the rat medial meniscal transection model. CJ-13610 at oral doses of 0.6, 2 and 6 mg/kg/day reversed two modalities of pain in this model; tactile allodynia and weight bearing differential. Taken together, these data suggest that 5-LOX pathway and the leukotriene products are important mediators of pain.

Novel Keto-phospholipids Are Generated by Monocytes and Macrophages, Detected in Cystic Fibrosis, and Activate Peroxisome Proliferator-activated Receptor-γ*

PubMed Central

Hammond, Victoria J.; Morgan, Alwena H.; Lauder, Sarah; Thomas, Christopher P.; Brown, Sarah; Freeman, Bruce A.; Lloyd, Clare M.; Davies, Jane; Bush, Andrew; Levonen, Anna-Liisa; Kansanen, Emilia; Villacorta, Luis; Chen, Y. Eugene; Porter, Ned; Garcia-Diaz, Yoel M.; Schopfer, Francisco J.; O'Donnell, Valerie B.

2012-01-01

12/15-Lipoxygenases (LOXs) in monocytes and macrophages generate novel phospholipid-esterified eicosanoids. Here, we report the generation of two additional families of related lipids comprising 15-ketoeicosatetraenoic acid (KETE) attached to four phosphatidylethanolamines (PEs). The lipids are generated basally by 15-LOX in IL-4-stimulated monocytes, are elevated on calcium mobilization, and are detected at increased levels in bronchoalveolar lavage fluid from cystic fibrosis patients (3.6 ng/ml of lavage). Murine peritoneal macrophages generate 12-KETE-PEs, which are absent in 12/15-LOX-deficient mice. Inhibition of 15-prostaglandin dehydrogenase prevents their formation from exogenous 15-hydroxyeicosatetraenoic acid-PE in human monocytes. Both human and murine cells also generated analogous hydroperoxyeicosatetraenoic acid-PEs. The electrophilic reactivity of KETE-PEs is shown by their Michael addition to glutathione and cysteine. Lastly, both 15-hydroxyeicosatetraenoic acid-PE and 15-KETE-PE activated peroxisome proliferator-activated receptor-γ reporter activity in macrophages in a dose-dependent manner. In summary, we demonstrate novel peroxisome proliferator-activated receptor-γ-activating oxidized phospholipids generated enzymatically by LOX and 15-prostaglandin dehydrogenase in primary monocytic cells and in a human Th2-related lung disease. The lipids are a new family of bioactive mediators from the 12/15-LOX pathway that may contribute to its known anti-inflammatory actions in vivo. PMID:23060450

Resistance to Fusarium verticillioides and fumonisin accumulation in maize inbred lines involves an earlier and enhanced expression of lipoxygenase (LOX) genes.

PubMed

Maschietto, Valentina; Marocco, Adriano; Malachova, Alexandra; Lanubile, Alessandra

2015-09-01

Fusarium verticillioides causes ear rot in maize and contaminates the kernels with the fumonisin mycotoxins. It is known that plant lipoxygenase (LOX)-derived oxylipins regulate defence against pathogens and that the host-pathogen lipid cross-talk influences the pathogenesis. The expression profiles of fifteen genes of the LOX pathway were studied in kernels of resistant and susceptible maize lines, grown in field condition, at 3, 7 and 14 days post inoculation (dpi) with F. verticillioides. Plant defence responses were correlated with the pathogen growth, the expression profiles of fungal FUM genes for fumonisin biosynthesis and fumonisin content in the kernels. The resistant genotype limited fungal growth and fumonisin accumulation between 7 and 14 dpi. Pathogen growth became exponential in the susceptible line after 7 dpi, in correspondence with massive transcription of FUM genes and fumonisins augmented exponentially at 14 dpi. LOX pathway genes resulted strongly induced after pathogen inoculation in the resistant line at 3 and 7 dpi, whilst in the susceptible line the induction was reduced or delayed at 14 dpi. In addition, all genes resulted overexpressed before infection in kernels of the resistant genotype already at 3 dpi. The results suggest that resistance in maize may depend on an earlier activation of LOX genes and genes for jasmonic acid biosynthesis. Copyright © 2015 Elsevier GmbH. All rights reserved.

Novel keto-phospholipids are generated by monocytes and macrophages, detected in cystic fibrosis, and activate peroxisome proliferator-activated receptor-γ.

PubMed

Hammond, Victoria J; Morgan, Alwena H; Lauder, Sarah; Thomas, Christopher P; Brown, Sarah; Freeman, Bruce A; Lloyd, Clare M; Davies, Jane; Bush, Andrew; Levonen, Anna-Liisa; Kansanen, Emilia; Villacorta, Luis; Chen, Y Eugene; Porter, Ned; Garcia-Diaz, Yoel M; Schopfer, Francisco J; O'Donnell, Valerie B

2012-12-07

12/15-Lipoxygenases (LOXs) in monocytes and macrophages generate novel phospholipid-esterified eicosanoids. Here, we report the generation of two additional families of related lipids comprising 15-ketoeicosatetraenoic acid (KETE) attached to four phosphatidylethanolamines (PEs). The lipids are generated basally by 15-LOX in IL-4-stimulated monocytes, are elevated on calcium mobilization, and are detected at increased levels in bronchoalveolar lavage fluid from cystic fibrosis patients (3.6 ng/ml of lavage). Murine peritoneal macrophages generate 12-KETE-PEs, which are absent in 12/15-LOX-deficient mice. Inhibition of 15-prostaglandin dehydrogenase prevents their formation from exogenous 15-hydroxyeicosatetraenoic acid-PE in human monocytes. Both human and murine cells also generated analogous hydroperoxyeicosatetraenoic acid-PEs. The electrophilic reactivity of KETE-PEs is shown by their Michael addition to glutathione and cysteine. Lastly, both 15-hydroxyeicosatetraenoic acid-PE and 15-KETE-PE activated peroxisome proliferator-activated receptor-γ reporter activity in macrophages in a dose-dependent manner. In summary, we demonstrate novel peroxisome proliferator-activated receptor-γ-activating oxidized phospholipids generated enzymatically by LOX and 15-prostaglandin dehydrogenase in primary monocytic cells and in a human Th2-related lung disease. The lipids are a new family of bioactive mediators from the 12/15-LOX pathway that may contribute to its known anti-inflammatory actions in vivo.

Study on atomization and combustion characteristics of LOX/methane pintle injectors

NASA Astrophysics Data System (ADS)

Fang, Xin-xin; Shen, Chi-bing

2017-07-01

Influences of main structural parameters of the LOX/methane pintle injectors on atomization cone angles and combustion performances were studied by experiments and numerical simulation respectively. In addition, improvement was brought up to the structure of the pintle injectors and combustion flow fields of two different pintle engines were obtained. The results indicate that, with increase of the gas-liquid mass flow ratio, the atomization cone angle decreases. In the condition of the same gas-liquid mass flow ratio, as the thickness of the LOX-injection gap grows bigger, the atomization cone angle becomes smaller. In the opposite, when the half cone angle of the LOX-injection gap grows bigger, the atomization cone angle becomes bigger. Moreover, owing to the viscous effects of the pintle tip, with increase of the 'skip distance', the atomization cone angle gets larger. Two big recirculation zones in the combustor lead to combustion stability of the pintle engines. When the value of the non-dimensional 'skip distance' is near 1, the combustion efficiency of the pintle engines is the highest. Additionally, pintle engines with LOX injected in quadrangular slots can acquire better mixing efficiency of the propellants and higher combustion efficiency as the gas methane can pass through the adjacent slots. However, the annular-channel type of pintle injectors has an 'enclosed' area near the pintle tip which has a great negative influence on the combustion efficiency.

Maternal transcription of non-protein coding RNAs from the PWS-critical region rescues growth retardation in mice.

PubMed

Rozhdestvensky, Timofey S; Robeck, Thomas; Galiveti, Chenna R; Raabe, Carsten A; Seeger, Birte; Wolters, Anna; Gubar, Leonid V; Brosius, Jürgen; Skryabin, Boris V

2016-02-05

Prader-Willi syndrome (PWS) is a neurogenetic disorder caused by loss of paternally expressed genes on chromosome 15q11-q13. The PWS-critical region (PWScr) contains an array of non-protein coding IPW-A exons hosting intronic SNORD116 snoRNA genes. Deletion of PWScr is associated with PWS in humans and growth retardation in mice exhibiting ~15% postnatal lethality in C57BL/6 background. Here we analysed a knock-in mouse containing a 5'HPRT-LoxP-Neo(R) cassette (5'LoxP) inserted upstream of the PWScr. When the insertion was inherited maternally in a paternal PWScr-deletion mouse model (PWScr(p-/m5'LoxP)), we observed compensation of growth retardation and postnatal lethality. Genomic methylation pattern and expression of protein-coding genes remained unaltered at the PWS-locus of PWScr(p-/m5'LoxP) mice. Interestingly, ubiquitous Snord116 and IPW-A exon transcription from the originally silent maternal chromosome was detected. In situ hybridization indicated that PWScr(p-/m5'LoxP) mice expressed Snord116 in brain areas similar to wild type animals. Our results suggest that the lack of PWScr RNA expression in certain brain areas could be a primary cause of the growth retardation phenotype in mice. We propose that activation of disease-associated genes on imprinted regions could lead to general therapeutic strategies in associated diseases.

Aviator’s Breathing Oxygen Specifications

DTIC Science & Technology

1976-12-01

Sheinson (2]), it can cocrystallize with acetylene. While solid acetylene will float on LOX, cocrystals with over 50% nitrous oxide can form a suspension...in LOX, and cocrystals wi’h over 70% nitrous oxide will sink and can detonate. Thus the simultatieous presence of icetylene and nitrous oxide above

Combustion performance and heat transfer characterization of LOX/hydrocarbon type propellants

NASA Technical Reports Server (NTRS)

Gross, R. S.

1980-01-01

A sound data base was established by analytically and experimentally generating basic regenerative cooling, combustion performance, combustion stability, and combustion chamber heat transfer parameters for LOX/HC propellants, with specific application to second generation orbit maneuvering and reaction control systems (OMS/RCS) for the Space Shuttle Orbiter.

The Lunar Occultation Explorer (LOX): Establishing the Moon as a Platform for Next-Generation Nuclear Astrophysics Investigations

NASA Astrophysics Data System (ADS)

Miller, R. S.; Ajello, M.; Beacom, J. F.; Bloser, P. F.; Burrows, A.; Errando, M.; Goldsten, J. O.; Hartmann, D.; Hoeflich, P.; Hungerford, A.; Lawrence, D. J.; Leary, J. C.; Leising, M. D.; Milne, P.; Peplowski, P. N.; The, L.-S.

2018-02-01

The Lunar Occultation Explorer (LOX) is a paradigm shift that will leverage the power of a new observational paradigm to transform our understanding of the nuclear cosmos (0.1-10 MeV) and establish the Moon as a platform for astrophysics.

Nutrient Loadings to Streams of the Continental United States from Municipal and Industrial Effluent

USGS Publications Warehouse

Maupin, M.A.; Ivahnenko, T.

2011-01-01

Data from the United States Environmental Protection Agency Permit Compliance System national database were used to calculate annual total nitrogen (TN) and total phosphorus (TP) loads to surface waters from municipal and industrial facilities in six major regions of the United States for 1992, 1997, and 2002. Concentration and effluent flow data were examined for approximately 118,250 facilities in 45 states and the District of Columbia. Inconsistent and incomplete discharge locations, effluent flows, and effluent nutrient concentrations limited the use of these data for calculating nutrient loads. More concentrations were reported for major facilities, those discharging more than 1million gallons per day, than for minor facilities, and more concentrations were reported for TP than for TN. Analytical methods to check and improve the quality of the Permit Compliance System data were used. Annual loads were calculated using "typical pollutant concentrations" to supplement missing concentrations based on the type and size of facilities. Annual nutrient loads for over 26,600 facilities were calculated for at least one of the three years. Sewage systems represented 74% of all TN loads and 58% of all TP loads. This work represents an initial set of data to develop a comprehensive and consistent national database of point-source nutrient loads. These loads can be used to inform a wide range of water-quality management, watershed modeling, and research efforts at multiple scales. ?? 2011 American Water Resources Association. This article is a U.S. Government work and is in the public domain in the USA.

Opportunities for Open Automated Demand Response in Wastewater Treatment Facilities in California - Phase II Report. San Luis Rey Wastewater Treatment Plant Case Study

DOE Office of Scientific and Technical Information (OSTI.GOV)

Thompson, Lisa; Lekov, Alex; McKane, Aimee

2010-08-20

This case study enhances the understanding of open automated demand response opportunities in municipal wastewater treatment facilities. The report summarizes the findings of a 100 day submetering project at the San Luis Rey Wastewater Treatment Plant, a municipal wastewater treatment facility in Oceanside, California. The report reveals that key energy-intensive equipment such as pumps and centrifuges can be targeted for large load reductions. Demand response tests on the effluent pumps resulted a 300 kW load reduction and tests on centrifuges resulted in a 40 kW load reduction. Although tests on the facility?s blowers resulted in peak period load reductions ofmore » 78 kW sharp, short-lived increases in the turbidity of the wastewater effluent were experienced within 24 hours of the test. The results of these tests, which were conducted on blowers without variable speed drive capability, would not be acceptable and warrant further study. This study finds that wastewater treatment facilities have significant open automated demand response potential. However, limiting factors to implementing demand response are the reaction of effluent turbidity to reduced aeration load, along with the cogeneration capabilities of municipal facilities, including existing power purchase agreements and utility receptiveness to purchasing electricity from cogeneration facilities.« less

Vehicle-Level Oxygen/Methane Propulsion System Hotfire Testing at Thermal Vacuum Conditions

NASA Technical Reports Server (NTRS)

Morehead, Robert L.; Melcher, J. C.; Atwell, Matthew J.; Hurlbert, Eric A.; Desai, Pooja; Werlink, Rudy

2017-01-01

A prototype integrated liquid oxygen/liquid methane propulsion system was hot-fire tested at a variety of simulated altitude and thermal conditions in the NASA Glenn Research Center Plum Brook Station In-Space Propulsion Thermal Vacuum Chamber (formerly B2). This test campaign served two purposes: 1) Characterize the performance of the Plum Brook facility in vacuum accumulator mode and 2) Collect the unique data set of an integrated LOX/Methane propulsion system operating in high altitude and thermal vacuum environments (a first). Data from this propulsion system prototype could inform the design of future spacecraft in-space propulsion systems, including landers. The test vehicle for this campaign was the Integrated Cryogenic Propulsion Test Article (ICPTA), which was constructed for this project using assets from the former Morpheus Project rebuilt and outfitted with additional new hardware. The ICPTA utilizes one 2,800 lbf main engine, two 28 lbf and two 7 lbf reaction control engines mounted in two pods, four 48-inch propellant tanks (two each for liquid oxygen and liquid methane), and a cold helium system for propellant tank pressurization. Several hundred sensors on the ICPTA and many more in the test cell collected data to characterize the operation of the vehicle and facility. Multiple notable experiments were performed during this test campaign, many for the first time, including pressure-fed cryogenic reaction control system characterization over a wide range of conditions, coil-on-plug ignition system demonstration at the vehicle level, integrated main engine/RCS operation, and a non-intrusive propellant mass gauging system. The test data includes water-hammer and thermal heat leak data critical to validating models for use in future vehicle design activities. This successful test campaign demonstrated the performance of the updated Plum Brook In-Space Propulsion thermal vacuum chamber and incrementally advanced the state of LOX/Methane propulsion technology through numerous system-level and subsystem experiments.

Advanced active health monitoring system of liquid rocket engines

NASA Astrophysics Data System (ADS)

Qing, Xinlin P.; Wu, Zhanjun; Beard, Shawn; Chang, Fu-Kuo

2008-11-01

An advanced SMART TAPE system has been developed for real-time in-situ monitoring and long term tracking of structural integrity of pressure vessels in liquid rocket engines. The practical implementation of the structural health monitoring (SHM) system including distributed sensor network, portable diagnostic hardware and dedicated data analysis software is addressed based on the harsh operating environment. Extensive tests were conducted on a simulated large booster LOX-H2 engine propellant duct to evaluate the survivability and functionality of the system under the operating conditions of typical liquid rocket engines such as cryogenic temperature, vibration loads. The test results demonstrated that the developed SHM system could survive the combined cryogenic temperature and vibration environments and effectively detect cracks as small as 2 mm.

The Use of a Block Diagram Simulation Language for Rapid Model Prototyping

NASA Technical Reports Server (NTRS)

Whitlow, Johnathan E.; Engrand, Peter

1996-01-01

The research performed this summer was a continuation of work performed during the 1995 NASA/ASEE Summer Fellowship. The focus of the work was to expand previously generated predictive models for liquid oxygen (LOX) loading into the external fuel tank of the shuttle. The models which were developed using a block diagram simulation language known as VisSim, were evaluated on numerous shuttle flights and found to well in most cases. Once the models were refined and validated, the predictive methods were integrated into the existing Rockwell software propulsion advisory tool (PAT). Although time was not sufficient to completely integrate the models developed into PAT, the ability to predict flows and pressures in the orbiter section and graphically display the results was accomplished.

Critical Surface Cleaning and Verification Alternatives

NASA Technical Reports Server (NTRS)

Melton, Donald M.; McCool, A. (Technical Monitor)

2000-01-01

As a result of federal and state requirements, historical critical cleaning and verification solvents such as Freon 113, Freon TMC, and Trichloroethylene (TCE) are either highly regulated or no longer 0 C available. Interim replacements such as HCFC 225 have been qualified, however toxicity and future phase-out regulations necessitate long term solutions. The scope of this project was to qualify a safe and environmentally compliant LOX surface verification alternative to Freon 113, TCE and HCFC 225. The main effort was focused on initiating the evaluation and qualification of HCFC 225G as an alternate LOX verification solvent. The project was scoped in FY 99/00 to perform LOX compatibility, cleaning efficiency and qualification on flight hardware.

Dual inhibitory activities of tannins from Hamamelis virginiana and related polyphenols on 5-lipoxygenase and lyso-PAF: acetyl-CoA acetyltransferase.

PubMed

Hartisch, C; Kolodziej, H; von Bruchhausen, F

1997-04-01

In the present study, the effects of tannins obtained from various plant sources on the activity of 5-LOX and AT were examined. With IC50 values ranging from 1.0 to 18.7 muM, hamamelitannin and the galloylated proanthocyanidins isolated from Hamamelis virginiana L. were found to be most potent inhibitors of 5-LOX. Unlike the 5-LOX study, hamamelitannin proved to be ineffective in the AT assay. Potent candidates are represented by the group of B-type proanthocyanidins. Structure-activity relationships regarding the in vitro inhibitory potency of the polyphenols in the biological assays are discussed.

Low-thrust Isp sensitivity study

NASA Technical Reports Server (NTRS)

Schoenman, L.

1982-01-01

A comparison of the cooling requirements and attainable specific impulse performance of engines in the 445 to 4448N thrust class utilizing LOX/RP-1, LOX/Hydrogen and LOX/Methane propellants is presented. The unique design requirements for the regenerative cooling of low-thrust engines operating at high pressures (up to 6894 kPa) were explored analytically by comparing single cooling with the fuel and the oxidizer, and dual cooling with both the fuel and the oxidizer. The effects of coolant channel geometry, chamber length, and contraction ratio on the ability to provide proper cooling were evaluated, as was the resulting specific impulse. The results show that larger contraction ratios and smaller channels are highly desirable for certain propellant combinations.

Maize death acids, 9-lipoxygenase derived cyclopente(a)nones, display activity as cytotoxic phytoalexins and transcriptional mediators

USDA-ARS?s Scientific Manuscript database

Plant damage promotes the interaction of lipoxygenases (LOX) with fatty acids yielding 9-hydroperoxides, 13-hydroperoxides and complex arrays of oxylipins. The action of 13-LOX on linolenic acid enables production of 12-oxo-phytodienoic acid (12-OPDA) and its downstream products, termed jasmonates. ...

Risk Assessment and scaling for the SLS LOx ET

NASA Technical Reports Server (NTRS)

Osipov, Viatcheslav V.; Hafiychuk, Halyna; Devine, Ekaterina V.; Khasin, Michael; Smelyanskiy, Vadim N.

2012-01-01

In this report we analyze the transpiration cooling by He bubble injection of the long LOx tank feedline heated by the environment heat. We consider possible hazards that can arise in the proposed design of the SLS core stage where the feedline length is much longer than that used in the Space Shuttle.

Application of the Cre/loxP Site-Specific Recombination System for Gene Transformation in Aurantiochytrium limacinum.

PubMed

Sun, Hengyi; Chen, Hao; Zang, Xiaonan; Hou, Pan; Zhou, Bingbing; Liu, Yuantao; Wu, Fei; Cao, Xiaofei; Zhang, Xuecheng

2015-06-01

The Cre/loxP site-specific recombination system was applied to Aurantiochytrium limacinum to obtain a transformant without the antibiotic resistance marker gene. First, the enhanced green fluorescent protein gene (egfp) and chloramphenicol resistance gene (Cmr), along with the two loxP loci, were integrated into the genome of A. limacinum OUC88 using 18S rDNA sequences as the homologous recombination sites. Then plasmid pSH65, containing a zeocin resistance gene (Bler) was transferred into A. limacinum OUC_CG. After induction with galactose, repeated passage in culture and PCR-based assessment, the pSH65 plasmid was lost and A. limacinum OUC_EG host was shown to no longer have resistance to 100 mg chloramphenicol/L or 5 mg zeocin/L. Through southern blotting and fluorescence detection, egfp was found to be integrated into the genome of A. limacinum OUC_EG, and EGFP was successfully expressed in the cells. The successful application of the Cre/loxP system demonstrates an experimental basis for genetic modification of A. limacinum so as to obtain transformed strains with no antibiotic resistance marker genes.

Dynamics of a Finite Liquid Oxygen (LOX) Column in a Pulsed Magnetic Field

NASA Technical Reports Server (NTRS)

Youngquist, Robert; Immer, Christopher; Lane, John; Simpson, James; Steinrock, T. (Technical Monitor)

2002-01-01

It is well known that liquid oxygen has a sufficient paramagnetic susceptibility that a strong magnetic field gradient can lift it in the earth's gravitational field. The movement of liquid oxygen is vital to the space program since it one of the primary oxidizers used for propulsion. Transport of liquid oxygen (LOX) via direct interaction of the magnetic fields (B field) with the fluid is a current topic of research and development at Kennedy Space Center, FL. This method of transporting (i.e. pumping) LOX may have particular advantages on Mars and other reduced gravitational environments, namely safety and reliability. This paper will address transport of a magnetic fluid, LOX, via phased-pulsed electromagnets acting on the edge of the column of fluid. The authors have developed a physical model from first-principles for the motion of a magnetic fluid in a particular U-tube geometry subjected to a pulsed magnetic field from an arbitrary solenoidal electromagnet. Experimental data that have been collected from the analogous geometry correlate well to that of the ab-initio calculations.

Contamination of liquid oxygen by pressurized gaseous nitrogen

NASA Technical Reports Server (NTRS)

Zuckerwar, Allan J.; King, Tracy K.; Ngo, Kim Chi

1989-01-01

The penetration of pressurized gaseous nitrogen (GN2) into liquid oxygen (LOX) was investigated experimentally in the 7-inch High Temperature Tunnel, the pilot tunnel for the 8-foot High Temperature Tunnel (8'HTT) at Langley Research Center. A preliminary test using a nuclear monitor revealed the extent of the liquid nitrogen (LN2) build-up at the LOX interface as a function of GN2 pressure. Then an adaptation of the differential flash vaporization technique was used to determine the binary diffusivity of the LOX-LN2 system at a temperature of 90.2 K. The measured value D equals 0.000086 sq cm/s + or - 25 percent together with two prior measurements at lower temperatures revealed an excellent fit to the Arrhenius equation, yielding a pre-exponential factor D sub 0 equals 0.0452 sq cm/s and an activation enthalpy H equals 1.08 kcal/mol. At a pressure of 1700 psi and holding time of 15 min, the penetration of LN2 into LOX (to a 1 percent contamination level) was found to be 0.9 cm, indicating but minimal impact upon 8'HTT operations.

Artesian Well Abandonment at Launch Complex 39A

NASA Technical Reports Server (NTRS)

Morgan, Lindsay; Johansen, Deda

2015-01-01

The artesian well tasked for abandonment was located on the LOX side (northwest area) of the launch complex. The exact date of well installation is unknown. The well was no longer in use at the time of the abandonment request, but was previously utilized under St. Johns River Water Management District (SJRWMD) consumptive use permit (No. 50054) for the Floridian Aquifer. The exact construction details of the LOX artesian well were also unknown; however, a similar-type artesian well was previously located on the LH2 side of the site, which was abandoned in 2012. Based on discussions with the NASA RPM and review of the LH2 artesian well abandonment completion report, the LH2 artesian well was reported to be an 8-inch diameter, 330-foot deep well. The NASA RPM communicated that the LOX artesian well was likely to be an 8-inch diameter, 380-foot deep well. This information was used for scoping, and was subsequently confirmed to be substantially accurate. No additional information could be found for the LOX artesian well using the NASA Remediation Information System (RIS).

International Space Exploration Coordination Group Assessment of Technology Gaps for LOx/Methane Propulsion Systems for the Global Exploration Roadmap

NASA Technical Reports Server (NTRS)

Hurlbert, Eric A.; Whitley, Ryan; Klem, Mark D.; Johnson, Wesley; Alexander, Leslie; D'Aversa, Emanuela; Ruault, Jean-Marc; Manfletti, Chiara; Caruana, Jean-Noel; Ueno, Hiroshi;

Release of lipoxygenase products and monoterpenes by tomato plants as an indicator of Botrytis cinerea-induced stress.

PubMed

Jansen, R M C; Miebach, M; Kleist, E; van Henten, E J; Wildt, J

2009-11-01

Changes in emission of volatile organic compounds (VOCs) from tomato induced by the fungus Botrytis cinerea were studied in plants inoculated by spraying with suspensions containing B. cinerea spores. VOC emissions were analysed using on-line gas chromatography-mass spectrometry, with a time resolution of about 1 h, for up to 2 days after spraying. Four phases were delimited according to the starting point and the applied day/night rhythm of the experiments. These phases were used to demonstrate changes in VOC flux caused by B. cinerea infestation. Tomato plants inoculated with B. cinerea emitted a different number and amount of VOCs after inoculation compared to control plants that had been sprayed with a suspension without B. cinerea spores. The changes in emissions were dependent on time after inoculation as well as on the severity of infection. The predominant VOCs emitted after inoculation were volatile products from the lipoxygenase pathway (LOX products). The increased emission of LOX products proved to be a strong indicator of a stress response, indicating that VOC emissions can be used to detect plant stress at an early stage. Besides emission of LOX products, there were also increases in monoterpene emissions. However, neither increased emission of LOX products nor of monoterpenes is specific for B. cinerea attack. The emission of LOX products is also induced by other stresses, and increased emission of monoterpenes seems to be the result of mechanical damage induced by secondary stress impacts on leaves.

Ursolic Acid Attenuates Atherosclerosis in ApoE-/- Mice: Role of LOX-1 Mediated by ROS/NF-κB Pathway.

PubMed

Li, Qiu; Zhao, Wenwen; Zeng, Xi; Hao, Zhihui

2018-05-07

Atherosclerosis, a chronic inflammatory disease, is a major contributor to cardiovascular diseases. Ursolic acid (UA) is a phytonutrient with widely biological effects including anti-oxidative, anti-inflammatory, and so on. At present, the effect of UA on atherosclerosis and the mechanism of action are still obscure. This study focused on investigating the effects of UA on atherosclerosis both in vivo and in vitro. We first selected LOX-1 as our target, which was reckoned as a new promising receptor for treating atherosclerosis. The evaluation in vitro suggested that UA significantly decreased endothelial LOX-1 expression induced by LPS both in mRNA and protein levels. Pre-treatment of UA also inhibited TLR4/MyD88 signaling activated by LPS. Moreover, UA reduced ROS production and suppressed the activation of NF-κB stimulated by LPS. Particularly, the evaluation in vivo further verified the conclusion obtained in vitro. In ApoE −/− mice fed with an atherogenic diet, both UA (100 mg/kg/day) and simvastatin significantly attenuated atherosclerotic plaque formation and shrunk necrotic core areas. The enhanced expression of LOX-1 in atherosclerotic aorta was also dramatically decreased by administration of UA. Taken together, these results suggested that UA, with anti-atherosclerotic activity through inhibition of LOX-1 mediated by ROS/NF-κB signaling pathways, may become a valuable vascular protective candidate for the treatment of atherosclerosis.

Transgene-free iPSCs generated from small volume peripheral blood nonmobilized CD34+ cells

PubMed Central

Merling, Randall K.; Sweeney, Colin L.; Choi, Uimook; De Ravin, Suk See; Myers, Timothy G.; Otaizo-Carrasquero, Francisco; Pan, Jason; Linton, Gilda; Chen, Lifeng; Koontz, Sherry; Theobald, Narda L.; Malech, Harry L.

2013-01-01

A variety of somatic cells can be reprogrammed to induced pluripotent stem cells (iPSCs), but CD34+ hematopoietic stem cells (HSCs) present in nonmobilized peripheral blood (PB) would be a convenient target. We report a method for deriving iPSC from PB HSCs using immunobead purification and 2- to 4-day culture to enrich CD34+ HSCs to 80% ± 9%, followed by reprogramming with loxP-flanked polycistronic (human Oct4, Klf4, Sox2, and c-Myc) STEMCCA-loxP lentivector, or with Sendai vectors. Colonies arising with STEMCCA-loxP were invariably TRA-1-60+, yielding 5.3 ± 2.8 iPSC colonies per 20 mL PB (n = 17), where most colonies had single-copy STEMCCA-loxP easily excised by transient Cre expression. Colonies arising with Sendai were variably reprogrammed (10%-80% TRA-1-60+), with variable yield (6 to >500 TRA-1-60+ iPSC colonies per 10 mL blood; n = 6). Resultant iPSC clones expressed pluripotent cell markers and generated teratomas. Genomic methylation patterns of STEMCCA-loxP–reprogrammed clones closely matched embryonic stem cells. Furthermore, we showed that iPSCs are derived from the nonmobilized CD34+ HSCs enriched from PB rather than from any lymphocyte or monocyte contaminants because they lack somatic rearrangements typical of T or B lymphocytes and because purified CD14+ monocytes do not yield iPSC colonies under these reprogramming conditions. PMID:23386128

The sunburn response in human skin is characterized by sequential eicosanoid profiles that may mediate its early and late phases.

PubMed

Rhodes, Lesley E; Gledhill, Karl; Masoodi, Mojgan; Haylett, Ann K; Brownrigg, Margaret; Thody, Anthony J; Tobin, Desmond J; Nicolaou, Anna

2009-11-01

Sunburn is a commonly occurring acute inflammatory process, with dermal vasodilatation and leukocyte infiltration as central features. Ultraviolet (UV) B-induced hydrolysis of membrane phospholipids releases polyunsaturated fatty acids, and their subsequent metabolism by cyclooxygenases (COXs) and lipoxygenases (LOXs) may produce potent eicosanoid mediators modulating different stages of the inflammation. Our objective was to identify candidate eicosanoids formed during the sunburn reaction in relation to its clinical and histological course. We exposed skin of healthy humans (n=32) to UVB and, for 72 h, examined expression of proinflammatory and anti-inflammatory eicosanoids using LC/ESI-MS/MS, and examined immunohistochemical expression of COX-2, 12-LOX, 15-LOX, and leukocyte markers, while quantifying clinical erythema. We show that vasodilatory prostaglandins (PGs) PGE(2), PGF(2alpha), and PGE(3) accompany the erythema in the first 24-48 h, associated with increased COX-2 expression at 24 h. Novel, potent leukocyte chemoattractants 11-, 12-, and 8-monohydroxy-eicosatetraenoic acid (HETE) are elevated from 4 to 72 h, in association with peak dermal neutrophil influx at 24 h, and increased dermal CD3(+) lymphocytes and 12- and 15-LOX expression from 24 to 72 h. Anti-inflammatory metabolite 15-HETE shows later expression, peaking at 72 h. Sunburn is characterized by overlapping sequential profiles of increases in COX products followed by LOX products that may regulate subsequent events and ultimately its resolution.

Maternal transcription of non-protein coding RNAs from the PWS-critical region rescues growth retardation in mice

PubMed Central

Rozhdestvensky, Timofey S.; Robeck, Thomas; Galiveti, Chenna R.; Raabe, Carsten A.; Seeger, Birte; Wolters, Anna; Gubar, Leonid V.; Brosius, Jürgen; Skryabin, Boris V.

2016-01-01

Prader-Willi syndrome (PWS) is a neurogenetic disorder caused by loss of paternally expressed genes on chromosome 15q11-q13. The PWS-critical region (PWScr) contains an array of non-protein coding IPW-A exons hosting intronic SNORD116 snoRNA genes. Deletion of PWScr is associated with PWS in humans and growth retardation in mice exhibiting ~15% postnatal lethality in C57BL/6 background. Here we analysed a knock-in mouse containing a 5′HPRT-LoxP-NeoR cassette (5′LoxP) inserted upstream of the PWScr. When the insertion was inherited maternally in a paternal PWScr-deletion mouse model (PWScrp−/m5′LoxP), we observed compensation of growth retardation and postnatal lethality. Genomic methylation pattern and expression of protein-coding genes remained unaltered at the PWS-locus of PWScrp−/m5′LoxP mice. Interestingly, ubiquitous Snord116 and IPW-A exon transcription from the originally silent maternal chromosome was detected. In situ hybridization indicated that PWScrp−/m5′LoxP mice expressed Snord116 in brain areas similar to wild type animals. Our results suggest that the lack of PWScr RNA expression in certain brain areas could be a primary cause of the growth retardation phenotype in mice. We propose that activation of disease-associated genes on imprinted regions could lead to general therapeutic strategies in associated diseases. PMID:26848093

Magnaporthe oryzae Induces the Expression of a MicroRNA to Suppress the Immune Response in Rice.

PubMed

Zhang, Xin; Bao, Yalin; Shan, Deqi; Wang, Zhihui; Song, Xiaoning; Wang, Zhaoyun; Wang, Jiansheng; He, Liqiang; Wu, Liang; Zhang, Zhengguang; Niu, Dongdong; Jin, Hailing; Zhao, Hongwei

2018-05-01

MicroRNAs play crucial roles in plant responses to pathogen infections. The rice blast disease, caused by the fungus Magnaporthe oryzae , is the most important disease of rice ( Oryza sativa ). To explore the microRNA species that participate in rice immunity against the rice blast disease, we compared the expression of small RNAs between mock- and M. oryzae -treated rice. We found that infection by M. oryzae strain Guy11 specifically induced the expression of rice miR319 and, consequently, suppressed its target gene TEOSINTE BRANCHED/CYCLOIDEA/PROLIFERATING CELL FACTOR1 ( OsTCP21 ), which encodes a transcription factor. Using transgenic rice that overexpresses miR319b (OE) or expresses OsTCP21 -Res (which is resistant to miR319-mediated silencing), we found that OsTCP21 is a positive regulator of the rice defense response against the blast disease. When wild-type and miR319b-OE rice were infected by Guy11, multiple jasmonic acid (JA) synthetic and signaling components were suppressed, indicating that Guy11 suppresses JA signaling through inducing miR319. In particular, we found that LIPOXYGENASE2 ( LOX2 ) and LOX5 were specifically suppressed by miR319 overexpression or by Guy11 infection. LOXs are the key enzymes of JA synthesis, which catalyze the conversion of α-linoleic acid to hydroperoxy-octadecadienoic acid. The application of α-linoleic acid rescued disease symptoms on the OsTCP21 -Res rice but not wild-type rice, supporting our hypothesis that OsLOX2 and OsLOX5 are the key JA synthesis genes hijacked by Guy11 to subvert host immunity and facilitate pathogenicity. We propose that induced expression of OsLOX2/5 may improve resistance to the rice blast disease. © 2018 American Society of Plant Biologists. All Rights Reserved.

Involvement of PLA2, COX and LOX in Rhinella arenarum oocyte maturation.

PubMed

Ortiz, Maria Eugenia; Bühler, Marta Inés; Zelarayán, Liliana Isabel

2014-11-01

In Rhinella arenarum, progesterone is the physiological nuclear maturation inducer that interacts with the oocyte surface and starts a cascade of events that leads to germinal vesicle breakdown (GVBD). Polyunsaturated fatty acids and their metabolites produced through cyclooxygenase (COX) and lipoxygenase (LOX) pathways play an important role in reproductive processes. In amphibians, to date, the role of arachidonic acid (AA) metabolites in progesterone (P4)-induced oocyte maturation has not been clarified. In this work we studied the participation of three enzymes involved in AA metabolism - phospholipase A2 (PLA2), COX and LOX in Rhinella arenarum oocyte maturation. PLA2 activation induced maturation in Rhinella arenarum oocytes in a dose-dependent manner. Oocytes when treated with 0.08 μM melittin showed the highest response (78 ± 6% GVBD). In follicles, PLA2 activation did not significantly induce maturation at the assayed doses (12 ± 3% GVBD). PLA2 inhibition with quinacrine prevented melittin-induced GVBD in a dose-dependent manner, however PLA2 inactivation did not affect P4-induced maturation. This finding suggests that PLA2 is not the only phospholipase involved in P4-induced maturation in this species. P4-induced oocyte maturation was inhibited by the COX inhibitors indomethacin and rofecoxib (65 ± 3% and 63 ± 3% GVBD, respectively), although COX activity was never blocked by their addition. Follicles showed a similar response following the addition of these inhibitors. Participation of LOX metabolites in maturation seems to be correlated with seasonal variation in ovarian response to P4. During the February to June period (low P4 response), LOX inhibition by nordihydroguaiaretic acid or lysine clonixinate increased maturation by up to 70%. In contrast, during the July to January period (high P4 response), LOX inhibition had no effect on hormone-induced maturation.

Natural Forms of Vitamin E and 13′-Carboxychromanol, a Long-Chain Vitamin E Metabolite, Inhibit Leukotriene Generation from Stimulated Neutrophils by Blocking Calcium Influx and Suppressing 5-Lipoxygenase Activity, Respectively

PubMed Central

Jiang, Ziying; Yin, Xinmin; Jiang, Qing

2014-01-01

Leukotrienes generated by 5-lipoxygenase (5-LOX)–catalyzed reaction are key regulators of inflammation. In ionophore-stimulated (A23187; 1–2.5 μM) human blood neutrophils or differentiated HL-60 cells, vitamin E forms differentially inhibited leukotriene B4 (LTB4) with an IC50 of 5–20 μM for γ-tocopherol, δ-tocopherol (δT), and γ-tocotrienol, but a much higher IC50 for α-tocopherol. 13′-Carboxychromanol, a long-chain metabolite of δT, suppressed neutrophil- and HL-60 cell-generated LTB4 with an IC50 of 4–7 μM and potently inhibited human recombinant 5-LOX activity with an IC50 of 0.5–1 μM. In contrast, vitamin E forms had no effect on human 5-LOX activity but impaired ionophore-induced intracellular calcium increase and calcium influx as well as the subsequent signaling including ERK1/2 phosphorylation and 5-LOX translocation from cytosol to the nucleus, a key event for 5-LOX activation. Further investigation showed that δT suppressed cytosolic Ca2+ increase and/or LTB4 formation triggered by ionophores, sphingosine 1-phosphate, and lysophosphatidic acid but not by fMLP or thapsigargin, whereas 13′-carboxychromanol decreased cellular production of LTB4 regardless of different stimuli, consistent with its strong inhibition of the 5-LOX activity. These observations suggest that δT does not likely affect fMLP receptor-mediated signaling or store depletion-induced calcium entry. Instead, we found that δT prevented ionophore-caused cytoplasmic membrane disruption, which may account for its blocking of calcium influx. These activities by vitamin E forms and long-chain carboxychromanol provide potential molecular bases for the differential anti-inflammatory effects of vitamin E forms in vivo. PMID:21169551

Formation of a Cyclopropyl Epoxide via a Leukotriene A Synthase-related Pathway in an Anaerobic Reaction of Soybean Lipoxygenase-1 with 15S-Hydroperoxyeicosatetraenoic Acid

PubMed Central

Zheng, Yuxiang; Brash, Alan R.

2010-01-01

The further conversion of an arachidonic acid hydroperoxide to a leukotriene A (LTA) type epoxide by specific lipoxygenase (LOX) enzymes constitutes a key step in inflammatory mediator biosynthesis. Whereas mammalian 5-LOX transforms its primary product (5S-hydroperoxyeicosatetraenoic acid; 5S-HPETE) almost exclusively to LTA4, the model enzyme, soybean LOX-1, normally produces no detectable leukotrienes and instead further oxygenates its primary product 15S-HPETE to 5,15- and 8,15-dihydroperoxides. Mammalian 15-LOX-1 displays both types of activity. We reasoned that availability of molecular oxygen within the LOX active site favors oxygenation, whereas lack of O2 promotes LTA epoxide synthesis. To test this, we reacted 15S-HPETE with soybean LOX-1 under anaerobic conditions and identified the products by high pressure liquid chromatography, UV, mass spectrometry, and NMR. Among the products, we identified a pair of 8,15-dihydroxy diastereomers with all-trans-conjugated trienes that incorporated 18O from H218O at C-8, indicative of the formation of 14,15-LTA4. A pair of 5,15-dihydroxy diastereomers containing two trans,trans-conjugated dienes (6E,8E,11E,13E) and that incorporated 18O from H218O at C-5 was deduced to arise from hydrolysis of a novel epoxide containing a cyclopropyl ring, 14,15-epoxy-[9,10,11-cyclopropyl]-eicosa-5Z,7E,13E-trienoic acid. Also identified was the δ-lactone of the 5,15-diol, a derivative that exhibited no 18O incorporation due to its formation by intramolecular reaction of the carboxyl anion with the proposed epoxide intermediate. Our results support a model in which access to molecular oxygen within the active site directs the outcome from competing pathways in the secondary reactions of lipoxygenases. PMID:20194505

Minimizing the effects of oxygen interference on l-lactate sensors by a single amino acid mutation in Aerococcus viridansl-lactate oxidase.

PubMed

Hiraka, Kentaro; Kojima, Katsuhiro; Lin, Chi-En; Tsugawa, Wakako; Asano, Ryutaro; La Belle, Jeffrey T; Sode, Koji

2018-04-30

l-lactate biosensors employing l-lactate oxidase (LOx) have been developed mainly to measure l-lactate concentration for clinical diagnostics, sports medicine, and the food industry. Some l-lactate biosensors employ artificial electron mediators, but these can negatively impact the detection of l-lactate by competing with the primary electron acceptor: molecular oxygen. In this paper, a strategic approach to engineering an AvLOx that minimizes the effects of oxygen interference on sensor strips was reported. First, we predicted an oxygen access pathway in Aerococcus viridans LOx (AvLOx) based on its crystal structure. This was subsequently blocked by a bulky amino acid substitution. The resulting Ala96Leu mutant showed a drastic reduction in oxidase activity using molecular oxygen as the electron acceptor and a small increase in dehydrogenase activity employing an artificial electron acceptor. Secondly, the Ala96Leu mutant was immobilized on a screen-printed carbon electrode using glutaraldehyde cross-linking method. Amperometric analysis was performed with potassium ferricyanide as an electron mediator under argon or atmospheric conditions. Under argon condition, the response current increased linearly from 0.05 to 0.5mM l-lactate for both wild-type and Ala96Leu. However, under atmospheric conditions, the response of wild-type AvLOx electrode was suppressed by 9-12% due to oxygen interference. The Ala96Leu mutant maintained 56-69% of the response current at the same l-lactate level and minimized the relative bias error to -19% from -49% of wild-type. This study provided significant insight into the enzymatic reaction mechanism of AvLOx and presented a novel approach to minimize oxygen interference in sensor applications, which will enable accurate detection of l-lactate concentrations. Copyright © 2017 Elsevier B.V. All rights reserved.

Magnaporthe oryzae Induces the Expression of a MicroRNA to Suppress the Immune Response in Rice1[OPEN

PubMed Central

Zhang, Xin; Bao, Yalin; Shan, Deqi; Wang, Zhihui; Song, Xiaoning; Wang, Zhaoyun; Wang, Jiansheng; He, Liqiang; Wu, Liang; Zhang, Zhengguang; Niu, Dongdong

2018-01-01

MicroRNAs play crucial roles in plant responses to pathogen infections. The rice blast disease, caused by the fungus Magnaporthe oryzae, is the most important disease of rice (Oryza sativa). To explore the microRNA species that participate in rice immunity against the rice blast disease, we compared the expression of small RNAs between mock- and M. oryzae-treated rice. We found that infection by M. oryzae strain Guy11 specifically induced the expression of rice miR319 and, consequently, suppressed its target gene TEOSINTE BRANCHED/CYCLOIDEA/PROLIFERATING CELL FACTOR1 (OsTCP21), which encodes a transcription factor. Using transgenic rice that overexpresses miR319b (OE) or expresses OsTCP21-Res (which is resistant to miR319-mediated silencing), we found that OsTCP21 is a positive regulator of the rice defense response against the blast disease. When wild-type and miR319b-OE rice were infected by Guy11, multiple jasmonic acid (JA) synthetic and signaling components were suppressed, indicating that Guy11 suppresses JA signaling through inducing miR319. In particular, we found that LIPOXYGENASE2 (LOX2) and LOX5 were specifically suppressed by miR319 overexpression or by Guy11 infection. LOXs are the key enzymes of JA synthesis, which catalyze the conversion of α-linoleic acid to hydroperoxy-octadecadienoic acid. The application of α-linoleic acid rescued disease symptoms on the OsTCP21-Res rice but not wild-type rice, supporting our hypothesis that OsLOX2 and OsLOX5 are the key JA synthesis genes hijacked by Guy11 to subvert host immunity and facilitate pathogenicity. We propose that induced expression of OsLOX2/5 may improve resistance to the rice blast disease. PMID:29549093

Level of tissue differentiation influences the activation of a heat-inducible flower-specific system for genetic containment in poplar (Populus tremula L.).

PubMed

Hoenicka, Hans; Lehnhardt, Denise; Nunna, Suneetha; Reinhardt, Richard; Jeltsch, Albert; Briones, Valentina; Fladung, Matthias

2016-02-01

Differentiation level but not transgene copy number influenced activation of a gene containment system in poplar. Heat treatments promoted CRE gene body methylation. The flower-specific transgene deletion was confirmed. Gene flow between genetic modified trees and their wild relatives is still motive of concern. Therefore, approaches for gene containment are required. In this study, we designed a novel strategy for achieving an inducible and flower-specific transgene removal from poplar trees but still expressing the transgene in the plant body. Hence, pollen carrying transgenes could be used for breeding purposes under controlled conditions in a first phase, and in the second phase genetic modified poplars developing transgene-free pollen grains could be released. This approach is based on the recombination systems CRE/loxP and FLP/frt. Both gene constructs contained a heat-inducible CRE/loxP-based spacer sequence for in vivo assembling of the flower-specific FLP/frt system. This allowed inducible activation of gene containment. The FLP/frt system was under the regulation of a flower-specific promoter, either CGPDHC or PTD. Our results confirmed complete CRE/loxP-based in vivo assembling of the flower-specific transgene excision system after heat treatment in all cells for up to 30 % of regenerants derived from undifferentiated tissue cultures. Degradation of HSP::CRE/loxP spacer after recombination but also persistence as extrachromosomal DNA circles were detected in sub-lines obtained after heat treatments. Furthermore, heat treatment promoted methylation of the CRE gene body. A lower methylation level was detected at CpG sites in transgenic sub-lines showing complete CRE/loxP recombination and persistence of CRE/loxP spacer, compared to sub-lines with incomplete recombination. However, our results suggest that low methylation might be necessary but not sufficient for recombination. The flower-specific FLP/frt-based transgene deletion was confirmed in 6.3 % of flowers.

Hepatocyte specific deletion of c-Met leads to the development of severe non-alcoholic steatohepatitis in mice.

PubMed

Kroy, Daniela C; Schumacher, Fabienne; Ramadori, Pierluigi; Hatting, Maximilian; Bergheim, Ina; Gassler, Nikolaus; Boekschoten, Mark V; Müller, Michael; Streetz, Konrad L; Trautwein, Christian

2014-10-01

Non-alcoholic-fatty-liver disease (NAFLD) is part of the metabolic syndrome. The spectrum of NAFLD includes NASH (non-alcoholic steatohepatitis), which is characterised by progressive inflammation associated with oxidative stress and apoptosis, finally triggering liver cirrhosis and hepatocellular carcinoma. HGF (hepatocyte growth factor)/mesenchymal-epithelial transition factor (c-Met) receptor signalling is known to activate distinct intracellular pathways mediating among others anti-apoptotic properties to hepatocytes. Therefore, the aim was to characterise the role of c-Met during NASH development. Hepatocyte specific c-Met knockout mice (c-MetΔ(hepa)) using the cre-loxP system and wild type controls (c-Met(loxP/loxP)) were fed a methionine-choline deficient (MCD) diet. MCD feeding triggered massive steatosis, decreased survival and higher transaminases in c-MetΔ(hepa) livers compared to c-Met(loxP/loxP). Gene array analysis demonstrated that genes involved in fatty acid metabolism were strongly upregulated in c-MetΔ(hepa) livers correlating with higher amounts of hepatic free fatty acids. Consequently, c-MetΔ(hepa) mice showed significantly more TUNEL positive cells and more superoxide anion production than c-Met(loxPloxP) animals. Additionally, c-MetΔ(hepa) livers showed significantly larger fractions of infiltrating neutrophils, macrophages, and cytotoxic T cells. These changes correlated with an enhanced progression of liver fibrosis as evidenced by higher collagen deposition in c-MetΔ(hepa) livers. As increased apoptosis was a prominent feature in c-MetΔ(hepa) livers, we generated c-Met/Casp8Δ(hepa) double knockout mice. In these animals compared to c-MetΔ(hepa) animals the increase in apoptosis could be reverted. c-Met deletion in hepatocytes triggers NASH progression. A prominent mechanism is higher fatty acid accumulation and increased apoptosis, which in part can be reverted by blocking caspase 8. Copyright © 2014 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Primary atomization of liquid jets issuing from rocket engine coaxial injectors

NASA Astrophysics Data System (ADS)

Woodward, Roger D.

1993-01-01

The investigation of liquid jet breakup and spray development is critical to the understanding of combustion phenomena in liquid-propellant rocket engines. Much work has been done to characterize low-speed liquid jet breakup and dilute sprays, but atomizing jets and dense sprays have yielded few quantitative measurements due to their optical opacity. This work focuses on a characteristic of the primary breakup process of round liquid jets, namely the length of the intact liquid core. The specific application considered is that of shear-coaxial type rocket engine injectors. Real-time x-ray radiography, capable of imaging through the dense two-phase region surrounding the liquid core, has been used to make the measurements. Nitrogen and helium were employed as the fuel simulants while an x-ray absorbing potassium iodide aqueous solution was used as the liquid oxygen (LOX) simulant. The intact-liquid-core length data have been obtained and interpreted to illustrate the effects of chamber pressure (gas density), injected-gas and liquid velocities, and cavitation. The results clearly show that the effect of cavitation must be considered at low chamber pressures since it can be the dominant breakup mechanism. A correlation of intact core length in terms of gas-to-liquid density ratio, liquid jet Reynolds number, and Weber number is suggested. The gas-to-liquid density ratio appears to be the key parameter for aerodynamic shear breakup in this study. A small number of hot-fire, LOX/hydrogen tests were also conducted to attempt intact-LOX-core measurements under realistic conditions in a single-coaxial-element rocket engine. The tests were not successful in terms of measuring the intact core, but instantaneous imaging of LOX jets suggests that LOX jet breakup is qualitatively similar to that of cold-flow, propellant-simulant jets. The liquid oxygen jets survived in the hot-fire environment much longer than expected, and LOX was even visualized exiting the chamber nozzle under some conditions. This may be an effect of the single element configuration.

Integrated Cryogenic Propulsion Test Article Thermal Vacuum Hotfire Testing

NASA Technical Reports Server (NTRS)

Morehead, Robert L.; Melcher, J. C.; Atwell, Matthew J.; Hurlbert, Eric A.

2017-01-01

In support of a facility characterization test, the Integrated Cryogenic Propulsion Test Article (ICPTA) was hotfire tested at a variety of simulated altitude and thermal conditions in the NASA Glenn Research Center Plum Brook Station In-Space Propulsion Thermal Vacuum Chamber (formerly B2). The ICPTA utilizes liquid oxygen and liquid methane propellants for its main engine and four reaction control engines, and uses a cold helium system for tank pressurization. The hotfire test series included high altitude, high vacuum, ambient temperature, and deep cryogenic environments, and several hundred sensors on the vehicle collected a range of system level data useful to characterize the operation of an integrated LOX/Methane spacecraft in the space environment - a unique data set for this propellant combination.

76 FR 39443 - National Environmental Policy Act; Santa Susana Field Laboratory

Federal Register 2010, 2011, 2012, 2013, 2014

2011-07-06

... analysis and consultation. These include surveys for wildlife, critical habitat, rare plants, wetlands, and..., Bravo, Coca, and Delta. Area II and the LOX Plant portion of Area I were acquired by NASA from the U.S... Area II and a portion of Area I (LOX Plant) of SSFL. As part of the environmental review process...

78 FR 47007 - National Environmental Policy Act; Santa Susana Field Laboratory

Federal Register 2010, 2011, 2012, 2013, 2014

2013-08-02

... consultation. These include surveys for wildlife, critical habitat, rare plants, wetlands, and archaeological... the LOX Plant portion of Area I were acquired by NASA from the U.S. Air Force in the 1970s. These test... groundwater remediation within Area II and a portion of Area I (Liquid Oxygen [LOX] Plant) of SSFL (pursuant...

78 FR 55763 - National Environmental Policy Act; Santa Susana Field Laboratory

Federal Register 2010, 2011, 2012, 2013, 2014

2013-09-11

... inform the analysis and consultation. These include surveys for wildlife, critical habitat, rare plants..., Coca, and Delta. Area II and the LOX Plant portion of Area I were acquired by NASA from the U.S. Air... Oxygen [LOX] Plant) of SSFL (pursuant to the 2007 Consent Order).'' As part of the environmental review...

Operational Issues in the Development of a Cost-Effective Reusable LOX/LH2 Engine

NASA Technical Reports Server (NTRS)

Ballard, Richard O.

2005-01-01

Contents include the following: SLI initiated under NASA Research Announcement (NRA) 8-30. Strategic Objectives. Make spaceflight safer (1 in 10000 mission LOV). Make spaceflight cheaper ($1000/lb payload). Two prototype LOX/LH2 engine systems funded under Cycle-1 of NRA8-30. COBRA (Pratt & Whitney / Aerojet). RS-83 (Rocketdyne).

Vessels installed at A-3

NASA Image and Video Library

2009-09-25

Construction of the A-3 Test Stand approaches another milestone with delivery and installation of water, isopropyl alcohol (IPA) and liquid oxygen (LOX) tanks. The three LOX tanks shown on the left and the two IPA tanks shown on the right are all 35,000 gallons each. The four water tanks in the center are 39,000 gallons each.

Over-expression of 12/15-lipoxygenase increases oxldl-induced pro-inflammatory mediator expression and foam cell formation in rodent macrophages

USDA-ARS?s Scientific Manuscript database

Blueberries (BB) prevent the development of atherosclerosis in apoE deficient mice, partly through suppressing 12/15-lipoxygenase (12/15-LOX) expression and reducing the production of oxidized lipid products. To further study the role of 12/15-LOX in atherosclerosis and investigate the inhibitory ef...

Maize death acids, 9-lipoxygenase-derived cyclopente(a)nones display activity as cytotoxic phytoalexins and transcriptional mediators

USDA-ARS?s Scientific Manuscript database

Plant cellular damage promotes the interaction of lipoxygenases (LOX) with free fatty acids to yield 9- and 13-hydroperoxides which are further metabolized into diverse oxylipins. The enzymatic action of 13-LOX on linolenic acid enables production of 12-oxo-phytodienoic acid (12-OPDA) and its downst...

The maize death acids, 10-oxo-11-phytoenoic acid and derivatives, demonstrate specificity in jasmonate-related signaling and defense

USDA-ARS?s Scientific Manuscript database

Plant cellular damage promotes the interaction of lipoxygenases (LOX) with free fatty acids to yield 9- and 13-hydroperoxides which are further metabolized into diverse oxylipins. The enzymatic action of 13-LOX on linolenic acid enables production of 12-oxo-phytodienoic acid (12-OPDA) and its downst...

Thermostable Lipoxygenase, a Key Enzyme in the Conversion of Linoleic Acid into Thrihydroxy-octadecenoic Acid by Pseudomonas aeruginosa PR3

USDA-ARS?s Scientific Manuscript database

Lipoxygenases (LOX) constitute a family of lipid-peroxidizing enzymes catalyzing the oxidation of unsaturated fatty acid with (1Z,4Z)-pentadiene structural unit, leading to formation of the conjugated (Z,E)-hydroperoxydienoic acid. LOXs have been known to be widely distributed in plants and animals...

Insulin autoimmune syndrome caused by an adhesive skin patch containing loxoprofen-sodium.

PubMed

Okazaki-Sakai, Satoko; Yoshimoto, Sachiko; Yagi, Kunimasa; Wakasugi, Takanobu; Takeda, Yoshiyu; Yamagishi, Masakazu

2013-01-01

A 62-year-old woman complained of repeated hypoglycemic events. A 75 g oral glucose tolerance test (75 gOGTT) showed a marked increase in the plasma insulin level and impaired glucose tolerance. The patient exhibited a high titer of plasma anti-insulin autoantibodies. Her diagnosis was insulin autoimmune syndrome (IAS). Following the cessation of loxoprofen-sodium (LOXs), she experienced no further hypoglycemic episodes. However, the hypoglycemic attacks recurred following the accidental readministration of LOXs in an adhesive skin patch. Considering the changes in the titer of anti-insulin autoantibodies, the repeated 75 gOGTT and the repeated Scatchard analysis, we determined LOXs to be the cause of the IAS and evaluated the characteristics of the autoantibodies.

Simultaneous and Sequential Integration by Cre/loxP Site-Specific Recombination in Saccharomyces cerevisiae.

PubMed

Choi, Ho-Jung; Kim, Yeon-Hee

2018-05-28

A Cre/ loxP -δ-integration system was developed to allow sequential and simultaneous integration of a multiple gene expression cassette in Saccharomyces cerevisiae . To allow repeated integrations, the reusable Candida glabrata MARKER ( CgMARKER ) carrying loxP sequences was used, and the integrated CgMARKER was efficiently removed by inducing Cre recombinase. The XYLP and XYLB genes encoding endoxylanase and β-xylosidase, respectively, were used as model genes for xylan metabolism in this system, and the copy number of these genes was increased to 15.8 and 16.9 copies/cell, respectively, by repeated integration. This integration system is a promising approach for the easy construction of yeast strains with enhanced metabolic pathways through multicopy gene expression.

Recent advances in lunar base simulation

NASA Astrophysics Data System (ADS)

Johenning, B.; Koelle, H. H.

This article reports about the results of the latest computer runs of a lunar base simulation model. The lunar base consists of 20 facilities for lunar mining, processing and fabrication. The infrastructure includes solar and nuclear power plants, a central workshop, habitat and farm. Lunar products can be used for construction of solar power systems (SPS) or other spacecraft at several space locations. The simulation model evaluates the mass, energy and manpower flows between the elements of the system as well as system cost and cost of products on an annual basis for a given operational period. The 1983 standard model run over a fifty-years life cycle (beginning about the year 2000) was accomplished for a mean annual production volume of 78 180 Mg of hardware products for export resulting in average specific manufacturing cost of 8.4 $/kg and total annual cost of 1.25 billion dollars during the life cycle. The reference space transportation system uses LOX/LH 2 propulsion for which at the average 210 500 Mg LOX per year is produced on the moon. The sensitivity analysis indicates the importance of bootstrapping as well as the influence of market size, space transportation cost and specific resources demand on the mean lunar manufacturing cost. The option using lunar resources turns out to be quite attractive from the economical viewpoint. Systems analysis by this lunar base model and further trade-offs will be a useful tool to confirm this.

Lipid Profiling of the Arabidopsis Hypersensitive Response Reveals Specific Lipid Peroxidation and Fragmentation Processes: Biogenesis of Pimelic and Azelaic Acid1[C][W

PubMed Central

Zoeller, Maria; Stingl, Nadja; Krischke, Markus; Fekete, Agnes; Waller, Frank; Berger, Susanne; Mueller, Martin J.

2012-01-01

Lipid peroxidation (LPO) is induced by a variety of abiotic and biotic stresses. Although LPO is involved in diverse signaling processes, little is known about the oxidation mechanisms and major lipid targets. A systematic lipidomics analysis of LPO in the interaction of Arabidopsis (Arabidopsis thaliana) with Pseudomonas syringae revealed that LPO is predominantly confined to plastid lipids comprising galactolipid and triacylglyceride species and precedes programmed cell death. Singlet oxygen was identified as the major cause of lipid oxidation under basal conditions, while a 13-lipoxygenase (LOX2) and free radical-catalyzed lipid oxidation substantially contribute to the increase upon pathogen infection. Analysis of lox2 mutants revealed that LOX2 is essential for enzymatic membrane peroxidation but not for the pathogen-induced free jasmonate production. Despite massive oxidative modification of plastid lipids, levels of nonoxidized lipids dramatically increased after infection. Pathogen infection also induced an accumulation of fragmented lipids. Analysis of mutants defective in 9-lipoxygenases and LOX2 showed that galactolipid fragmentation is independent of LOXs. We provide strong in vivo evidence for a free radical-catalyzed galactolipid fragmentation mechanism responsible for the formation of the essential biotin precursor pimelic acid as well as of azelaic acid, which was previously postulated to prime the immune response of Arabidopsis. Our results suggest that azelaic acid is a general marker for LPO rather than a general immune signal. The proposed fragmentation mechanism rationalizes the pathogen-induced radical amplification and formation of electrophile signals such as phytoprostanes, malondialdehyde, and hexenal in plastids. PMID:22822212

Lipid profiling of the Arabidopsis hypersensitive response reveals specific lipid peroxidation and fragmentation processes: biogenesis of pimelic and azelaic acid.

PubMed

Zoeller, Maria; Stingl, Nadja; Krischke, Markus; Fekete, Agnes; Waller, Frank; Berger, Susanne; Mueller, Martin J

2012-09-01

Lipid peroxidation (LPO) is induced by a variety of abiotic and biotic stresses. Although LPO is involved in diverse signaling processes, little is known about the oxidation mechanisms and major lipid targets. A systematic lipidomics analysis of LPO in the interaction of Arabidopsis (Arabidopsis thaliana) with Pseudomonas syringae revealed that LPO is predominantly confined to plastid lipids comprising galactolipid and triacylglyceride species and precedes programmed cell death. Singlet oxygen was identified as the major cause of lipid oxidation under basal conditions, while a 13-lipoxygenase (LOX2) and free radical-catalyzed lipid oxidation substantially contribute to the increase upon pathogen infection. Analysis of lox2 mutants revealed that LOX2 is essential for enzymatic membrane peroxidation but not for the pathogen-induced free jasmonate production. Despite massive oxidative modification of plastid lipids, levels of nonoxidized lipids dramatically increased after infection. Pathogen infection also induced an accumulation of fragmented lipids. Analysis of mutants defective in 9-lipoxygenases and LOX2 showed that galactolipid fragmentation is independent of LOXs. We provide strong in vivo evidence for a free radical-catalyzed galactolipid fragmentation mechanism responsible for the formation of the essential biotin precursor pimelic acid as well as of azelaic acid, which was previously postulated to prime the immune response of Arabidopsis. Our results suggest that azelaic acid is a general marker for LPO rather than a general immune signal. The proposed fragmentation mechanism rationalizes the pathogen-induced radical amplification and formation of electrophile signals such as phytoprostanes, malondialdehyde, and hexenal in plastids.

Omega-3 fatty acids are oxygenated at the n-7 carbon by the lipoxygenase domain of a fusion protein in the cyanobacterium Acaryochloris marina

PubMed Central

Gao, Benlian; Boeglin, William E.; Brash, Alan R.

2009-01-01

Lipoxygenases (LOX) are found in most organisms that contain polyunsaturated fatty acids, usually existing as individual genes although occasionally encoded as a fusion protein with a catalase-related hemoprotein. Such a fusion protein occurs in the cyanobacterium Acaryochloris marina and herein we report the novel catalytic activity of its LOX domain. The full-length protein and the C-terminal LOX domain were expressed in Escherichia coli, and the catalytic activities characterized by UV, HPLC, GC-MS, and CD. All omega-3 polyunsaturates were oxygenated by the LOX domain at the n-7 position and with R stereospecificity: α-linolenic and the most abundant fatty acid in A. marina, stearidonic acid (C18.4ω3), are converted to the corresponding 12R-hydroperoxides, eicosapentaenoic acid to its 14R-hydroperoxide, and docosahexaenoic acid to its 16R-hydroperoxide. Omega-6 polyunsaturates were oxygenated at the n-10 position, forming 9R-hydroperoxy-octadecadienoic acid from linoleic acid and 11R-hydroperoxy-eicosatetraenoic acid from arachidonic acid. The metabolic transformation of stearidonic acid by the full-length fusion protein entails its 12R oxygenation with subsequent conversion by the catalase-related domain to a novel allene epoxide, a likely precursor of cyclopentenone fatty acids or other signaling molecules (Gao et al, J. Biol. Chem. 284:22087-98, 2009). Although omega-3 fatty acids and lipoxygenases are of widespread occurrence, this appears to be the first description of a LOX-catalyzed oxygenation that specifically utilizes the terminal pentadiene of omega-3 fatty acids. PMID:19786119

Selective uptake and efflux of cholesteryl linoleate in LDL by macrophages expressing 12/15-lipoxygenase

DOE Office of Scientific and Technical Information (OSTI.GOV)

Takahashi, Yoshitaka; Zhu, Hong; Xu, Wanpeng

Oxidation of low density lipoprotein (LDL) is a critical step for airtightness, and the role of the 12/15-lipoxygenase (12/15-Lox) as well as LDL receptor-related protein (Lp) expressed in macrophages in this process has been suggested. The oxygenation of cholesteryl linoleate in LDL by mouse macrophage-like Joe.1 cells over expressing 12/15-Lox was inhibited by an anti-Lp antibody but not by an anti-LDL receptor antibody. When the cells were incubated with LDL double-labeled by [{sup 3}H]cholesteryl linoleate and [{sup 125}I]apo B, association with the cells of [{sup 3}H]cholesteryl linoleate expressed as LDL protein equivalent exceeded that of [{sup 125}I]apo B, indicating selectivemore » uptake of [{sup 3}H]cholesteryl linoleate from LDL to these cells. An anti-Lp antibody inhibited the selective uptake of [{sup 3}H]cholesteryl ester by 62% and 81% with the 12/15-Lox-expressing cells and macrophages, respectively. Furthermore, addition of LDL to the culture medium of the [{sup 3}H]cholesteryl linoleate-labeled 12/15-Lox-expressing cells increased the release of [{sup 3}H]cholesteryl linoleate to the medium in LDL concentration- and time-dependent manners. The transport of [{sup 3}H]cholesteryl linoleate from the cells to LDL was also inhibited by an anti-Lp antibody by 75%. These results strongly suggest that Lp contributes to the LDL oxidation by 12/15-Lox in macrophages by selective uptake and efflux of cholesteryl ester in the LDL particle.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xu, Guang-Lin; Department of Pharmacology, University of Michigan, Ann Arbor; Du, Yi-Fang

KYKZL-1, a newly synthesized compound with COX/5-LOX dual inhibition, was subjected to the anti-inflammatory activity test focusing on its modulation of inflammatory mediators as well as intracellular MAPK and NF-κB signaling pathways. In acute ear edema model, pretreatment with KYKZL-1 (p.o.) dose-dependently inhibited the xylene-induced ear edema in mice with a higher inhibition than diclofenac. In a three-day TPA-induced inflammation, KYKZL-1 also showed significant anti-inflammatory activity with inhibition ranging between 20% and 64%. In gastric lesion test, KYKZL-1 elicited markedly fewer stomach lesions with a low index of ulcer as compared to diclofenac in rats. In further studies, KYKZL-1 wasmore » found to significantly inhibit the production of NO, PGE{sub 2}, LTB{sub 4} in LPS challenged RAW264.7, which is parallel to its attenuation of the expression of iNOS, COX-2, 5-LOX mRNAs or proteins and inhibition of phosphorylation of p38 and ERK MAPKs and activation of NF-κB. Taken together, our data indicate that KYKZL-1 comprises dual inhibition of COX and 5-LOX and exerts an obvious anti-inflammatory activity with an enhanced gastric safety profile via simultaneous inhibition of phosphorylation of p38 and ERK MAPKs and activation of NF-κB. - Highlights: • KYKZL-1 is designed to exhibit COX/5-LOX dual inhibition. • KYKZL-1 inhibits NO, PGE{sub 2} and LTB{sub 4} and iNOS, COX-2 and 5-LOX mRNAs and MAPKs. • KYKZL-1 inhibits phosphorylation of MAPKs. • KYKZL-1 inactivates NF-κB pathway.« less

A simple performance calculation method for LH2/LOX engines with different power cycles

NASA Technical Reports Server (NTRS)

Schmucker, R. H.

1973-01-01

A simple method for the calculation of the specific impulse of an engine with a gas generator cycle is presented. The solution is obtained by a power balance between turbine and pump. Approximate equations for the performance of the combustion products of LH2/LOX are derived. Performance results are compared with solutions of different engine types.

LOx / LCH4: A Unifying Technology for Future Exploration

NASA Technical Reports Server (NTRS)

Falker, John; Terrier, Douglas; Clayton, Ronald G.; Banker, Brian; Ryan, Abigail

2015-01-01

Reduced mass due to increasing commonality between spacecraft subsystems such as power and propulsion have been identified as critical to enabling human missions to Mars. This project represents the first ever integrated propulsion and power system testing and lays the foundations for future sounding rocket flight testing, which will yield the first in-space ignition of a LOx / LCH4 rocket engine.

LOX/Methane Main Engine Glow Plug Igniter Tests and Modeling

NASA Technical Reports Server (NTRS)

Breisacher, Kevin; Ajmani, Kumud

2009-01-01

Ignition data for tests with a LOX/methane igniter that utilized a glow plug as the ignition source are presented. The tests were conducted in a vacuum can with thermally conditioned (cold) hardware. Data showing the effects of glow plug geometry, type, and igniter operating conditions are discussed. Comparisons between experimental results and multidimensional, transient computer models are also made.

Influence of LOX/COX inhibitors on cell differentiation induced by all-trans retinoic acid in neuroblastoma cell lines.

PubMed

Redova, Martina; Chlapek, Petr; Loja, Tomas; Zitterbart, Karel; Hermanova, Marketa; Sterba, Jaroslav; Veselska, Renata

2010-02-01

We investigated the possible modulation by LOX/ COX inhibitors of all-trans retinoic acid (ATRA)-induced cell differentiation in two established neuroblastoma cell lines, SH-SY5Y and SK-N-BE(2). Caffeic acid, as an inhibitor of 5-lipoxygenase, and celecoxib, as an inhibitor of cyclooxygenase-2, were chosen for this study. The effects of the combined treatment with ATRA and LOX/COX inhibitors on neuroblastoma cells were studied using cell morphology assessment, detection of differentiation markers by immunoblotting, measurement of proliferation activity, and cell cycle analysis and apoptosis detection by flow cytometry. The results clearly demonstrated the potential of caffeic acid to enhance ATRA-induced cell differentiation, especially in the SK-N-BE(2) cell line, whereas application of celecoxib alone or with ATRA led predominantly to cytotoxic effects in both cell lines. Moreover, the higher sensitivity of the SK-N-BE(2) cell line to combined treatment with ATRA and LOX/COX inhibitors suggests that cancer stem cells are a main target for this therapeutic approach. Nevertheless, further detailed study of the phenomenon of enhanced cell differentiation by expression profiling is needed.

Summary of LOX/CH4 Thruster Technology Development at NASA/MSFC

NASA Technical Reports Server (NTRS)

Greene, Sandra Elam

2015-01-01

In recent years, a variety of injectors for liquid oxygen (LOX) and methane (CH4) propellant systems have been designed, fabricated, and demonstrated with hot-fire testing at Marshall Space Flight Center (MSFC). Successful designs for liquid methane (LCH4) and gaseous methane (GCH4) have been developed. A variety of chambers, including a transpiration cooled design, along with uncooled ablatives and refractory metals, have also been hot-fire tested by MSFC for use with LOX/LCH4 injectors. Hot-fire testing has also demonstrated multiple ignition source options. Heat flux data for selected injectors has been gathered by testing with a calorimeter chamber. High performance and stable combustion have been demonstrated, along with designs for thrust levels ranging from 500 to 7,000 lbf. The newest LOX/CH4 injector and chamber developed by MSFC have been fabricated with additive manufacturing techniques and include unique design features to investigate regenerative cooling with methane. This low cost and versatile hardware offers a design for 4,000 lbf thrust and will be hot-fire tested at MSFC in 2015. Its design and operation can easily be scaled for use in systems with thrust levels up to 25,000 lbf.

Refolding and characterization of the functional ligand-binding domain of human lectin-like oxidized LDL receptor.

PubMed

Xie, Qiuhong; Matsunaga, Shigeru; Shi, Xiaohua; Ogawa, Setsuko; Niimi, Setsuko; Wen, Zhesheng; Tokuyasu, Ken; Machida, Sachiko

2003-11-01

Lectin-like oxidized low-density lipoprotein receptor (LOX-1), a type II membrane protein that can recognize a variety of structurally unrelated macromolecules, plays an important role in host defense and is implicated in atherogenesis. To understand the interaction between human LOX-1 and its ligands, in this study the functional C-type lectin-like domain (CTLD) of LOX-1 was reconstituted at high efficiency from inactive aggregates in Escherichia coli using a refolding technique based on an artificial chaperone. The CD spectra of the purified domain suggested that the domain has alpha-helical structure and the blue shift of Trp residues was observed on refolding of the domain. Like wild-type hLOX-1, the refolded CTLD domain was able to bind modified LDL. Thus, even though CTLD contains six Cys residues that form disulfide bonds, it recovered its specific binding ability on refolding. This suggests that the correct disulfide bonds in CTLD were formed by the artificial chaperone technique. Although the domain lacked N-glycosylation, it showed high affinity for its ligand in surface plasmon resonance experiments. Thus, unglycosylated CTLD is sufficient for binding modified LDL.

Simulation of water vapor condensation on LOX droplet surface using liquid nitrogen

NASA Technical Reports Server (NTRS)

Powell, Eugene A.

1988-01-01

The formation of ice or water layers on liquid oxygen (LOX) droplets in the Space Shuttle Main Engine (SSME) environment was investigated. Formulation of such ice/water layers is indicated by phase-equilibrium considerations under conditions of high partial pressure of water vapor (steam) and low LOX droplet temperature prevailing in the SSME preburner or main chamber. An experimental investigation was begun using liquid nitrogen as a LOX simulant. A monodisperse liquid nitrogen droplet generator was developed which uses an acoustic driver to force the stream of liquid emerging from a capillary tube to break up into a stream of regularly space uniformly sized spherical droplets. The atmospheric pressure liquid nitrogen in the droplet generator reservoir was cooled below its boiling point to prevent two phase flow from occurring in the capillary tube. An existing steam chamber was modified for injection of liquid nitrogen droplets into atmospheric pressure superheated steam. The droplets were imaged using a stroboscopic video system and a laser shadowgraphy system. Several tests were conducted in which liquid nitrogen droplets were injected into the steam chamber. Under conditions of periodic droplet formation, images of 600 micron diameter liquid nitrogen droplets were obtained with the stroboscopic video systems.

[Effects of cadmium stress on fatty acid composition and lipid peroxidation of Malus hupehensis].

PubMed

You, Shu-Zhen; Yang, Hong-Qiang; Zhang, Long; Shao, Xiao-Jie

2009-08-01

This paper studied the fatty acid composition, reactive oxygen species (ROS), lipoxygenase (LOX) activity, and malondialdehyde (MDA) content in the leaves and roots of Malus hupehensis seedlings under effects of cadmium (Cd) stress. Noticeable changes were observed in the kinds and relative contents of fatty acids after treated with CdCl2 for 7-12 hours. The relative contents of unsaturated fatty acids in leaves and roots reached the maximum after treated for 7 hours, being 82. 82% and 72. 43% , respectively. The kinds of fatty acids in leaves increased from 11 to 14 after treated for 12 hours, while those in roots increased from 4 to 6 after treated for 17 hours. The O2* generation rate and the H2O2 content reached the maximum after treated for 3 and 7 hours, respectively, and the MDA content and LOX activity increased with treating time. Cd stress altered the fatty acid composition of Malus hupehensis via the inducement of reactive oxygen species and lipoxygenase, and induced lipid peroxidation, which was caused by both ROS and LOX within the first 12 hours of CdCl2 treatment and mainly by the increase of LOX activity since then.

Thrombin-activated human platelets acutely generate oxidized docosahexaenoic-acid-containing phospholipids via 12-lipoxygenase.

PubMed

Morgan, Lloyd T; Thomas, Christopher P; Kühn, Hartmut; O'Donnell, Valerie B

2010-10-01

Arachidonate-containing oxidized phospholipids are acutely generated by 12-LOX (12-lipoxygenase) in agonist-activated platelets. In the present study, formation of structurally related lipids by oxidation of DHA (docosahexaenoic acid)-containing phospholipids is demonstrated using lipidomic approaches. Precursor scanning reverse-phase LC (liquid chromatography)-MS/MS (tandem MS) identified a new family of lipids that comprise phospholipid-esterified HDOHE (hydroxydocosahexaenoic acid). Two diacyl and two plasmalogen PEs (phosphatidylethanolamines) containing predominantly the 14-HDOHE positional isomer (18:0p/14-HDOHE-PE, 18:0a/14-HDOHE-PE, 16:0a/14-HDOHE-PE and 16:0p/14-HDOHE-PE) were structurally characterized using MS/MS and by comparison with biogenic standards. An involvement of 12-LOX was indicated as purified recombinant human 12-LOX also generated the 14-HDOHE isomer from DHA. Pharmacological studies using inhibitors and recombinant platelet 12-LOX indicate that they form via esterification of newly formed non-esterified HDOHE. HDOHE-PEs formed at significant rates (2-4 ng/4×10(7) cells) within 2-180 min of thrombin stimulation, and their formation was blocked by calcium chelation. In summary, a new family of oxidized phospholipid was identified in thrombin-activated human platelets.

Liquid oxygen sloshing in Space Shuttle External Tank

NASA Technical Reports Server (NTRS)

Kannapel, M. D.; Przekwas, A. J.; Singhal, A. K.; Costes, N. C.

1987-01-01

This paper describes a numerical simulation of the hydrodynamics within the liquid oxygen tank of the Space Shuttle External Tank during liftoff. Before liftoff, the tank is filled with liquid oxygen (LOX) to approximately 97 percent with the other 3 percent containing gaseous oxygen (GOX) and helium. During liftoff, LOX is drained from the bottom of the tank, and GOX is pumped into the tank's ullage volume. There is a delay of several seconds before the GOX reaches the tank which causes the ullage pressure to decrease for several seconds after liftoff; this pressure 'slump' is a common phenomenon in rocket propulsion. When four slosh baffles were removed from the tank, the ullage gas pressure dropped more rapidly than in all previous flights. The purpose of this analysis was to determine whether the removal of the baffles could have caused the increased pressure 'slump' by changing the LOX surface dynamics. The results show that the LOX surface undergoes very high vertical accelerations (up to 5 g) and, therefore, splashing almost certainly occurs. The number of baffles does not affect the surface if the structural motion is assumed; but, the number of baffles may affect the structural motion of the tank.

Modification to the Langley 8-foot high temperature tunnel for hypersonic propulsion testing

NASA Technical Reports Server (NTRS)

Reubush, D. E.; Puster, R. L.; Kelly, H. N.

1987-01-01

Described are the modifications currently under way to the Langley 8-Foot High Temperature Tunnel to produce a new, unique national resource for testing hypersonic air-breathing propulsion systems. The current tunnel, which has been used for aerothermal loads and structures research since its inception, is being modified with the addition of a LOX system to bring the oxygen content of the test medium up to that of air, the addition of alternate Mach number capability (4 and 5) to augment the current M=7 capability, improvements to the tunnel hardware to reduce maintenance downtime, the addition of a hydrogen system to allow the testing of hydrogen powered engines, and a new data system to increase both the quantity and quality of the data obtained.

Bearing tester data compilation, analysis and reporting and bearing math modeling, volume 1

NASA Technical Reports Server (NTRS)

Marshall, D. D.; Montgomery, E. E.; New, L. S.; Stone, M. A.; Tiller, B. K.

1984-01-01

Thermal and mechanical models of high speed angular contact ball bearings operating in LOX and LN2 were developed and verified with limited test data in an effort to further understand the parameters that determine or effect the SSME turbopump bearing operational characteristics and service life. The SHABERTH bearing analysis program which was adapted to evaluate shaft bearing systems in cryogenics is not capable of accommodating varying thermal properties and two phase flow. A bearing model with this capability was developed using the SINDA thermal analyzer. Iteration between the SHABERTH and the SINDA models enable the establishment of preliminary bounds for stable operation in LN2. These limits were established in terms of fluid flow, fluid inlet temperature, and axial load for a shaft speed of 30,000 RPM.

Extreme winds and tornadoes: an overview

DOE Office of Scientific and Technical Information (OSTI.GOV)

McDonald, J.R.

1985-01-01

The objective of this course on extreme winds, hurricanes and tornadoes is to provide an overview of these natural phenomenon from the perspective of design of new buildings and structures or the evaluation of existing ones. Information is directly applicable to design and evaluation processes. The premise is that the facility under consideration, which may consist of various buildings, structures, processing equipment, stacks, ventilation ducts, etc., can be classified into certain categories, depending on the importance of the mission performed in the facility or the hazard that is presented by the particular operation. Having classified the facility into an appropriatemore » category will automatically define certain design goals for the facility. The design goals are then met by selecting a design wind speed that is appropriate for the specified exceedance probability and by following certain specified design procedures. The problem then is to determine appropriate wind loads and other applicable loads, including dead loads, live loads, seismic loads and other loads that may act on the structures. The design process can then proceed in the usual manner. In the case of existing facilities the strengths of the various structural elements, subsystems and systems are evaluated and these strengths are related to wind speeds that would result in failure to meet the design goals. 12 refs.« less

Liquid oxygen tank installed at A-3 Test Stand

NASA Technical Reports Server (NTRS)

2009-01-01

A liquid oxygen (LOX) tank is lifted into place at the A-3 Test Stand being built at NASA's John C. Stennis Space Center. Fourteen LOX, isopropyl alcohol (IPA) and water tanks are being installed to support the chemical steam generators to be used on the A-3 Test Stand. The IPA and LOX tanks will provide fuel for the generators. The water will allow the generators to produce steam that will be used to reduce pressure inside the stand's test cell diffuser, enabling operators to simulate altitudes up to 100,000 feet. In that way, operators can perform the tests needed on rocket engines being built to carry humans back to the moon and possibly beyond. The A-3 Test Stand is set for completion and activation in 2011.

LOX droplet vaporization in a supercritical forced convective environment

NASA Technical Reports Server (NTRS)

Hsiao, Chia-Chun; Yang, Vigor

1993-01-01

Modern liquid rocket engines often use liquid oxygen (LOX) and liquid hydrogen (LH2) as propellants to achieve high performance, with the engine operational conditions in the supercritical regimes of the propellants. Once the propellant exceeds its critical state, it essentially becomes a puff of dense fluid. The entire field becomes a continuous medium, and no distinct interfacial boundary between the liquid and gas exists. Although several studies have been undertaken to investigate the supercritical droplet behavior at quiescent conditions, very little effort has been made to address the fundamental mechanisms associated with LOX droplet vaporization in a supercritical, forced convective environment. The purpose is to establish a theoretical framework within which supercritical droplet dynamics and vaporization can be studied systematically by means of an efficient and robust numerical algorithm.

Liquid oxygen tank installed at A-3 Test Stand

NASA Image and Video Library

2009-09-18

A liquid oxygen (LOX) tank is lifted into place at the A-3 Test Stand being built at NASA's John C. Stennis Space Center. Fourteen LOX, isopropyl alcohol (IPA) and water tanks are being installed to support the chemical steam generators to be used on the A-3 Test Stand. The IPA and LOX tanks will provide fuel for the generators. The water will allow the generators to produce steam that will be used to reduce pressure inside the stand's test cell diffuser, enabling operators to simulate altitudes up to 100,000 feet. In that way, operators can perform the tests needed on rocket engines being built to carry humans back to the moon and possibly beyond. The A-3 Test Stand is set for completion and activation in 2011.

High-Pressure Oxygen Generation for Outpost EVA Study

NASA Technical Reports Server (NTRS)

Jeng, Frank F.; Conger, Bruce; Ewert, Michael K.; Anderson, Molly S.

2009-01-01

The amount of oxygen consumption for crew extravehicular activity (EVA) in future lunar exploration missions will be significant. Eight technologies to provide high pressure EVA O2 were investigated. They are: high pressure O2 storage, liquid oxygen (LOX) storage followed by vaporization, scavenging LOX from Lander followed by vaporization, LOX delivery followed by sorption compression, water electrolysis followed by compression, stand-alone high pressure water electrolyzer, Environmental Control and Life Support System (ECLSS) and Power Elements sharing a high pressure water electrolyzer, and ECLSS and In-Situ Resource Utilization (ISRU) Elements sharing a high pressure electrolyzer. A trade analysis was conducted comparing launch mass and equivalent system mass (ESM) of the eight technologies in open and closed ECLSS architectures. Technologies considered appropriate for the two architectures were selected and suggested for development.

Cycle O(CY1991) NLS trade studies and analyses report. Book 2, part 2: Propulsion

NASA Technical Reports Server (NTRS)

Cronin, R.; Werner, M.; Bonson, S.; Spring, R.; Houston, R.

1992-01-01

This report documents the propulsion system tasks performed in support of the National Launch System (NLS) Cycle O preliminary design activities. The report includes trades and analyses covering the following subjects: (1) Maximum Tank Stretch Study; (2) No LOX Bleed Performance Analysis; (3) LOX Bleed Trade Study; (4) LO2 Tank Pressure Limits; (5) LOX Tank Pressurization System Using Helium; (6) Space Transportation Main Engine (STME) Heat Exchanger Performance; (7) LH2 Passive Recirculation Performance Analysis; (8) LH2 Bleed/Recirculation Study; (9) LH2 Tank Pressure Limits; and (10) LH2 Pressurization System. For each trade study an executive summary and a detailed trade study are provided. For the convenience of the reader, a separate section containing a compilation of only the executive summaries is also provided.

NASA Langley's Aircraft Landing Dynamics Facility

NASA Technical Reports Server (NTRS)

Davis, Pamela A.

1993-01-01

The Aircraft Landing Dynamics Facility (ALDF) is a unique facility with the ability to test aircraft landing gear systems on actual runway surfaces at operational ground speeds and loading conditions. A brief historical overview of the original Landing Loads Track (LLT) is given, followed by a detailed description of the new ALDF systems and operational capabilities.

The NASA landing gear test airplane

NASA Technical Reports Server (NTRS)

Carter, John F.; Nagy, Christopher J.

1995-01-01

A tire and landing gear test facility has been developed and incorporated into a Convair 990 aircraft. The system can simulate tire vertical load profiles to 250,000 lb, sideslip angles to 15 degrees, and wheel braking on actual runways. Onboard computers control the preprogrammed test profiles through a feedback loop and also record three axis loads, tire slip angle, and tire condition. The aircraft to date has provided tire force and wear data for the Shuttle Orbiter tire on three different runways and at east and west coast landing sites. This report discusses the role of this facility in complementing existing ground tire and landing gear test facilities, and how this facility can simultaneously simulate the vertical load, tire slip, velocity, and surface for an entire aircraft landing. A description is given of the aircraft as well as the test system. An example of a typical test sequence is presented. Data collection and reduction from this facility are discussed, as well as accuracies of calculated parameters. Validation of the facility through ground and flight tests is presented. Tests to date have shown that this facility can operate at remote sites and gather complete data sets of load, slip, and velocity on actual runway surfaces. The ground and flight tests have led to a successful validation of this test facility.

Arachidonate 15-Lipoxygenase 2 as an Endogenous Inhibitor of Prostate Cancer Development

DTIC Science & Technology

2006-03-01

dehydrogenase; NHP, normal human prostate epithelial cells; PCa, prostate cancer; NLS, nuclear localization signal; PPAR -, peroxisome proliferator...cloned, i.e., 15-LOX2sv-a/b/c, are mostly excluded from the nucleus. A potential bi-partite nuclear localization signal (NLS...only partially involved in the nuclear import of 15-LOX2. To elucidate the relationship between nuclear localization , enzymatic activity, and tumor

Selective Production of 9R-Hydroxy-10E,12Z,15Z-Octadecatrienoic Acid from α-Linolenic Acid in Perilla Seed Oil Hydrolyzate by a Lipoxygenase from Nostoc Sp. SAG 25.82.

PubMed

Kim, Kyoung-Rok; An, Jung-Ung; Lee, Seon-Hwa; Oh, Deok-Kun

2015-01-01

Hydroxy fatty acids (HFAs) derived from omega-3 polyunsaturated fatty acids have been known as versatile bioactive molecules. However, its practical production from omega-3 or omega-3 rich oil has not been well established. In the present study, the stereo-selective enzymatic production of 9R-hydroxy-10E,12Z,15Z-octadecatrienoic acid (9R-HOTE) from α-linolenic acid (ALA) in perilla seed oil (PO) hydrolyzate was achieved using purified recombinant 9R-lipoxygenase (9R-LOX) from Nostoc sp. SAG 25.82. The specific activity of the enzyme followed the order linoleic acid (LA) > ALA > γ-linolenic acid (GLA). A total of 75% fatty acids (ALA and LA) were used as a substrate for 9R-LOX from commercial PO by hydrolysis of Candida rugosa lipase. The optimal reaction conditions for the production of 9R-HOTE from ALA using 9R-LOX were pH 8.5, 15°C, 5% (v/v) acetone, 0.2% (w/v) Tween 80, 40 g/L ALA, and 1 g/L enzyme. Under these conditions, 9R-LOX produced 37.6 g/L 9R-HOTE from 40 g/L ALA for 1 h, with a conversion yield of 94% and a productivity of 37.6 g/L/h; and the enzyme produced 34 g/L 9R-HOTE from 40 g/L ALA in PO hydrolyzate for 1 h, with a conversion yields of 85% and a productivity of 34 g/L/h. The enzyme also converted 9R-hydroxy-10E,12Z-octadecadienoic acid (9R-HODE) from 40 g/L LA for 1.0 h, with a conversion yield of 95% and a productivity of 38.4 g/L. This is the highest productivity of HFA from both ALA and ALA-rich vegetable oil using LOX ever reported. Therefore, our result suggests an efficient method for the production of 9R-HFAs from LA and ALA in vegetable oil using recombinant LOX in biotechnology.

Selective Production of 9R-Hydroxy-10E,12Z,15Z-Octadecatrienoic Acid from α-Linolenic Acid in Perilla Seed Oil Hydrolyzate by a Lipoxygenase from Nostoc Sp. SAG 25.82

PubMed Central

Kim, Kyoung-Rok; An, Jung-Ung; Lee, Seon-Hwa; Oh, Deok-Kun

2015-01-01

Hydroxy fatty acids (HFAs) derived from omega-3 polyunsaturated fatty acids have been known as versatile bioactive molecules. However, its practical production from omega-3 or omega-3 rich oil has not been well established. In the present study, the stereo-selective enzymatic production of 9R-hydroxy-10E,12Z,15Z-octadecatrienoic acid (9R-HOTE) from α-linolenic acid (ALA) in perilla seed oil (PO) hydrolyzate was achieved using purified recombinant 9R-lipoxygenase (9R-LOX) from Nostoc sp. SAG 25.82. The specific activity of the enzyme followed the order linoleic acid (LA) > ALA > γ-linolenic acid (GLA). A total of 75% fatty acids (ALA and LA) were used as a substrate for 9R-LOX from commercial PO by hydrolysis of Candida rugosa lipase. The optimal reaction conditions for the production of 9R-HOTE from ALA using 9R-LOX were pH 8.5, 15°C, 5% (v/v) acetone, 0.2% (w/v) Tween 80, 40 g/L ALA, and 1 g/L enzyme. Under these conditions, 9R-LOX produced 37.6 g/L 9R-HOTE from 40 g/L ALA for 1 h, with a conversion yield of 94% and a productivity of 37.6 g/L/h; and the enzyme produced 34 g/L 9R-HOTE from 40 g/L ALA in PO hydrolyzate for 1 h, with a conversion yields of 85% and a productivity of 34 g/L/h. The enzyme also converted 9R-hydroxy-10E,12Z-octadecadienoic acid (9R-HODE) from 40 g/L LA for 1.0 h, with a conversion yield of 95% and a productivity of 38.4 g/L. This is the highest productivity of HFA from both ALA and ALA-rich vegetable oil using LOX ever reported. Therefore, our result suggests an efficient method for the production of 9R-HFAs from LA and ALA in vegetable oil using recombinant LOX in biotechnology. PMID:26379279

Bypass of lethality with mosaic mice generated by Cre-loxP-mediated recombination.

PubMed

Betz, U A; Vosshenrich, C A; Rajewsky, K; Müller, W

1996-10-01

The analysis of gene function based on the generation of mutant mice by homologous recombination in embryonic stem cells is limited if gene disruption results in embryonic lethality. Mosaic mice, which contain a certain proportion of mutant cells in all organs, allow lethality to be circumvented and the potential of mutant cells to contribute to different cell lineages to be analyzed. To generate mosaic animals, we used the bacteriophage P1-derived Cre-loxP recombination system, which allows gene alteration by Cre-mediated deletion of loxP-flanked gene segments. We generated nestin-cre transgenic mouse lines, which expressed the Cre recombinase under the control of the rat nestin promoter and its second intron enhancer. In crosses to animals carrying a loxP-flanked target gene, partial deletion of the loxP-flanked allele occurred before day 10.5 post coitum and was detectable in all adult organs examined, including germ-line cells. Using this approach, we generated mosaic mice containing cells deficient in the gamma-chain of the interleukin-2 receptor (IL-2R gamma); in these animals, the IL-2R gamma-deficient cells were underrepresented in the thymus and spleen. Because mice deficient in DNA polymerase beta die perinatally, we studied the effects of DNA polymerase beta deficiency in mosaic animals. We found that some of the mosaic polymerase beta-deficient animals were viable, but were often reduced in size and weight. The fraction of DNA polymerase beta-deficient cells in mosaic embryos decreased during embryonic development, presumably because wild-type cells had a competitive advantage. The nestin-cre transgenic mice can be used to generate mosaic animals in which target genes are mutated by Cre-mediated recombination of loxP-flanked target genes. By using mosaic animals, embryonic lethality can be bypassed and cell lineages for whose development a given target gene is critical can be identified. In the case of DNA polymerase beta, deficient cells are already selected against during embryonic development, demonstrating the general importance of this protein in multiple cell types.

Design, Activation, and Operation of the J2-X Subscale Simulator (JSS)

NASA Technical Reports Server (NTRS)

Saunders, Grady P.; Raines, Nickey G.; Varner, Darrel G.

2009-01-01

The purpose of this paper is to give a detailed description of the design, activation, and operation of the J2-X Subscale Simulator (JSS) installed in Cell 1 of the E3 test facility at Stennis Space Center, MS (SSC). The primary purpose of the JSS is to simulate the installation of the J2-X engine in the A3 Subscale Rocket Altitude Test Facility at SSC. The JSS is designed to give aerodynamically and thermodynamically similar plume properties as the J2-X engine currently under development for use as the upper stage engine on the ARES I and ARES V spacecraft. The JSS is a scale pressure fed, LOX/GH fueled rocket that is geometrically similar to the J2-X from the throat to the nozzle exit plane (NEP) and is operated at the same oxidizer to fuel ratios and chamber pressures. This paper describes the heritage hardware used as the basis of the JSS design, the newly designed rocket hardware, igniter systems used, and the activation and operation of the JSS.

RS-88 Pad Abort Demonstrator Thrust Chamber Assembly Testing at NASA Marshall Space Flight Center

NASA Technical Reports Server (NTRS)

Farr, Rebecca A.; Sanders, Timothy M.

1990-01-01

This paper documents the effort conducted to collect hot-tire dynamic and acoustics environments data during 50,000-lb thrust lox-ethanol hot-fire rocket testing at NASA Marshall Space Flight Center (MSFC) in November-December 2003. This test program was conducted during development testing of the Boeing Rocketdyne RS-88 development engine thrust chamber assembly (TCA) in support of the Orbital Space Plane (OSP) Crew Escape System Propulsion (CESP) Program Pad Abort Demonstrator (PAD). In addition to numerous internal TCA and nozzle measurements, induced acoustics environments data were also collected. Provided here is an overview of test parameters, a discussion of the measurements, test facility systems and test operations, and a quality assessment of the data collected during this test program.

Differential modulation of leukotriene B4 synthesis and degradation in human bronchoalveolar lavage cells by lipopolysaccharide and tobacco smoke.

PubMed

Mao, Jenny T; Tashkin, Donald P; Tsu, I-Hsien; Serio, Kenneth J

2008-09-01

Leukotrienes have been implicated to play a prominent inductive role in carcinogenesis. We previously reported that bronchoalveolar lavage (BAL) cells from smokers manifested higher levels of leukotriene B4 (LTB4) production than ex-smokers. This study aims to elucidate the underlying mechanism(s). BAL cells from current and former smokers were exposed to lipopolysaccharide (LPS) for up to 7 days. LPS induced the release of LTB4 from BAL cells and down-regulated 5-lipoxygenase (5-LOX) mRNA expression in a dose-dependent manner, followed by a decrease in 5-LOX protein production and normalization of LTB4 levels. Exogenous LTB4 inhibited LPS-induced 5-LOX activity and accentuated the down-regulation of 5-LOX mRNA, whereas suppression of 5-LOX abrogated the LPS-induced changes, suggesting a negative feedback mechanism. LPS concomitantly induced expression and activity of the LTB4 metabolizing enzyme LTB4 omega-hydroxylase (LTB4OH) in ex-smokers' BAL cells, but not in smokers' BAL cells. In vitro smoke exposure of ex-smokers' BAL cells also abrogated the LPS-induced up-regulation of LTB4OH mRNA expression. Furthermore, ex-smokers' BAL cells expressed significantly higher LTB4OH mRNA levels than smokers' BAL cells. Such differential modulation of LTB4 synthesis and degradation by LPS in the setting of tobacco smoke exposure suggests that mechanisms responsible for sustained elevation of LTB4 levels in the lung microenvironment may contribute to the pathogenesis of tobacco-related respiratory diseases such as lung cancer. By regulating the balance of LTB4 in the lung, LTB4OH may function as a suppressor of lung carcinogenesis.

P2Y2R activation by nucleotides released from the highly metastatic breast cancer cell contributes to pre-metastatic niche formation by mediating lysyl oxidase secretion, collagen crosslinking, and monocyte recruitment

PubMed Central

Joo, Young Nak; Jin, Hana; Eun, So Young; Park, Sang Won; Chang, Ki Churl; Kim, Hye Jung

2014-01-01

Tumor microenvironmental hypoxia induces hypoxia inducible factor-1α (HIF-1α) overexpression, leading to the release of lysyl oxidase (LOX), which crosslinks collagen at distant sites to facilitate environmental changes that allow cancer cells to easily metastasize. Our previous study showed that activation of the P2Y2 receptor (P2Y2R) by ATP released from MDA-MB-231 cells increased MDA-MB-231 cell invasion through endothelial cells. Therefore, in this study, we investigated the role of P2Y2R in breast cancer cell metastasis to distant sites. ATP or UTP released from hypoxia-treated MDA-MB-231 cells induced HIF-1α expression and LOX secretion by the activation of P2Y2R, and this phenomenon was significantly reduced in P2Y2R-depleted MDA-MB-231 cells. Furthermore, P2Y2R-mediated LOX release induced collagen crosslinking in an in vitro model. Finally, nude mice injected with MDA-MB-231 cells showed high levels of LOX secretion, crosslinked collagen and CD11b+ BMDC recruitment in the lung; however, mice that were injected with P2Y2R-depleted MDA-MB-231 cells did not exhibit these changes. These results demonstrate that P2Y2R plays an important role in activation of the HIF-1α–LOX axis, the induction of collagen crosslinking and the recruitment of CD11b+ BMDCs. Furthermore, P2Y2R activation by nucleotides recruits THP-1 monocytes, resulting in primary tumor progression and pre-metastatic niche formation. PMID:25238333

TGF-beta1 induces the different expressions of lysyl oxidases and matrix metalloproteinases in anterior cruciate ligament and medial collateral ligament fibroblasts after mechanical injury.

PubMed

Xie, Jing; Wang, Chunli; Huang, Dong-Yue; Zhang, Yanyan; Xu, Jianwen; Kolesnikov, Stanislav S; Sung, K L Paul; Zhao, Hucheng

2013-03-15

The anterior cruciate ligament (ACL) is known to have a poor self-healing ability. In contrast, the medial collateral ligament (MCL) can heal relatively well and restore the joint function. Transforming growth factor-beta1 (TGF-β1) is considered to be an important chemical mediator in the wound healing of the ligaments. While the role of TGF-β1-induced expressions of the lysyl oxidases (LOXs) and matrix metalloproteinases (MMPs), which respectively facilitate the extracellular matrix (ECM) repair and degradation, is poorly understood. In this study, we used equibiaxial stretch chamber to mimic mechanical injury of ACL and MCL fibroblasts, and aimed to determine the intrinsic differences between ACL and MCL by characterizing the differential expressions of LOXs and MMPs in response to TGF-β1 after mechanical injury. By using semi-quantitative PCR, quantitative real-time PCR, western blot and zymography, we found TGF-β1 induced injured MCL to express more LOXs than injured ACL (up to 1.85-fold in LOX, 2.21-fold in LOXL-1, 1.71-fold in LOXL-2, 2.52-fold in LOXL-3 and 3.32-fold in LOXL-4). Meanwhile, TGF-β1 induced injured ACL to express more MMPs than injured MCL fibroblasts (up to 2.33-fold in MMP-1, 2.45-fold in MMP-2, 1.89-fold in MMP-3 and 1.50-fold in MMP-12). The further protein results were coincident with the gene expressions above. The different expressions of LOXs and MMPs inferred the intrinsic differences between ACL and MCL, and the intrinsic differences could help to explain their differential healing abilities. Copyright © 2012 Elsevier Ltd. All rights reserved.

Marker-free transgenic rice expressing the vegetative insecticidal protein (Vip) of Bacillus thuringiensis shows broad insecticidal properties.

PubMed

Pradhan, Subrata; Chakraborty, Anirban; Sikdar, Narattam; Chakraborty, Saikat; Bhattacharyya, Jagannath; Mitra, Joy; Manna, Anulina; Dutta Gupta, Snehasish; Sen, Soumitra Kumar

2016-10-01

Genetically engineered rice lines with broad insecticidal properties against major lepidopteran pests were generated using a synthetic, truncated form of vegetative insecticidal protein (Syn vip3BR) from Bacillus thuringiensis. The selectable marker gene and the redundant transgene(s) were eliminated through Cre/ lox mediated recombination and genetic segregation to make consumer friendly Bt -rice. For sustainable resistance against lepidopteran insect pests, chloroplast targeted synthetic version of bioactive core component of a vegetative insecticidal protein (Syn vip3BR) of Bacillus thuringiensis was expressed in rice under the control of green-tissue specific ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit gene promoter. The transgenic plants (in Oryza sativa indica Swarna cultivar) showed high insect mortality rate in vitro against major rice pests, yellow stem borer (Scirpophaga incertulas), rice leaf folder (Cnaphalocrocis medinalis) and rice horn caterpillar (Melanitis leda ismene) in T1 generation, indicating insecticidal potency of Syn vip3BR. Under field conditions, the T1 plants showed considerable resistance against leaf folders and stem borers. The expression cassette (vip-lox-hpt-lox) as well as another vector with chimeric cre recombinase gene under constitutive rice ubiquitin1 gene promoter was designed for the elimination of selectable marker hygromycin phosphotransferase (hptII) gene. Crossing experiments were performed between T1 plants with single insertion site of vip-lox-hpt-lox T-DNA and one T1 plant with moderate expression of cre recombinase with linked bialaphos resistance (syn bar) gene. Marker gene excision was achieved in hybrids with up to 41.18 % recombination efficiency. Insect resistant transgenic lines, devoid of selectable marker and redundant transgene(s) (hptII + cre-syn bar), were established in subsequent generation through genetic segregation.

Lysyl Oxidase-like-2 (LOXL2) Is a Major Isoform in Chondrocytes and Is Critically Required for Differentiation*

PubMed Central

Iftikhar, Mussadiq; Hurtado, Paola; Bais, Manish V.; Wigner, Nate; Stephens, Danielle N.; Gerstenfeld, Louis C.; Trackman, Philip C.

2011-01-01

The lysyl oxidase family is made up of five members: lysyl oxidase (LOX) and lysyl oxidase-like 1–4 (LOXL1-LOXL4). All members share conserved C-terminal catalytic domains that provide for lysyl oxidase or lysyl oxidase-like enzyme activity; and more divergent propeptide regions. LOX family enzyme activities catalyze the final enzymatic conversion required for the formation of normal biosynthetic collagen and elastin cross-links. The importance of lysyl oxidase enzyme activity to normal bone development has long been appreciated, but regulation and roles for specific LOX isoforms in bone formation in vivo is largely unexplored. Fracture healing recapitulates aspects of endochondral bone development. The present study first investigated the expression of all LOX isoforms in fracture healing. A remarkable coincidence of LOXL2 expression with the chondrogenic phase of fracture healing was found, prompting more detailed analyses of LOXL2 expression in normal growth plates, and LOXL2 expression and function in developing ATDC5 chondrogenic cells. Data show that LOXL2 is expressed by pre-hypertrophic and hypertrophic chondrocytes in vivo, and that LOXL2 expression is regulated in vitro as a function of chondrocyte differentiation. Moreover, LOXL2 knockdown studies in vitro show that LOXL2 expression is required for ATDC5 chondrocyte cell line differentiation through regulation of SNAIL and SOX9, important transcription factors that control chondrocyte differentiation. Taken together, data provide evidence that LOXL2, like LOX, is a multifunctional protein. LOXL2 promotes chondrocyte differentiation by mechanisms that are likely to include roles as both a regulator and an effector of chondrocyte differentiation. PMID:21071451

Flavocoxid, a dual inhibitor of cyclooxygenase-2 and 5-lipoxygenase, reduces pancreatic damage in an experimental model of acute pancreatitis

PubMed Central

Polito, F; Bitto, A; Irrera, N; Squadrito, F; Fazzari, C; Minutoli, L; Altavilla, D

2010-01-01

BACKGROUND AND PURPOSE Acute pancreatitis is an autodigestive process resulting in acute inflammation of the pancreas. Accumulating evidence indicates the essential contribution of cyclooxygenase (COX)-2 and 5-lipoxygenase (5-LOX) to acute pancreatitis. We studied the effects of flavocoxid, a plant-derived dual inhibitor of COX-2 and 5-LOX, in a model of caerulein (CER)-induced acute pancreatitis. EXPERIMENTAL APPROACH Rats were given CER (80 µg·kg−1 for each of four injections at hourly intervals) or vehicle (Sham-CER). Animals were then randomized to receive flavocoxid (20 mg·kg−1 i.p.) or vehicle, 30 min after the first CER injection. Two hours after the last CER injection, we evaluated damage to the pancreas by histological methods; serum levels of amylase, lipase, leukotriene (LT)B4 and prostaglandin (PG)E2; pancreatic expression of COX-2 and 5-LOX and tumour necrosis factor-α (TNF-α) gene expression by real-time polymerase chain reaction. KEY RESULTS Caerulein induced inflammatory changes in the pancreas and raised values of the other variables measured. In CER-treated animals, but not in those given saline, flavocoxid inhibited COX-2 and 5-LOX expression, reduced serum levels of lipase and amylase and the degree of pancreatic oedema. Treatment with flavocoxid blunted the increased pancreatic TNF-α mRNA expression, serum leukotriene B4 and prostaglandin E2 levels, and protected against histological damage in terms of vacuolization and leukocyte infiltration. CONCLUSIONS AND IMPLICATIONS Our results confirm the key role of both COX-2 and 5-LOX in the inflammatory response to acute pancreatitis. Flavocoxid may provide a potential therapeutic approach to the treatment of patients at high risk of developing this life-threatening condition. PMID:20977452

Flavocoxid, a dual inhibitor of cyclooxygenase-2 and 5-lipoxygenase, reduces pancreatic damage in an experimental model of acute pancreatitis.

PubMed

Polito, F; Bitto, A; Irrera, N; Squadrito, F; Fazzari, C; Minutoli, L; Altavilla, D

2010-11-01

Acute pancreatitis is an autodigestive process resulting in acute inflammation of the pancreas. Accumulating evidence indicates the essential contribution of cyclooxygenase (COX)-2 and 5-lipoxygenase (5-LOX) to acute pancreatitis. We studied the effects of flavocoxid, a plant-derived dual inhibitor of COX-2 and 5-LOX, in a model of caerulein (CER)-induced acute pancreatitis. Rats were given CER (80 µg·kg⁻¹ for each of four injections at hourly intervals) or vehicle (Sham-CER). Animals were then randomized to receive flavocoxid (20 mg·kg⁻¹ i.p.) or vehicle, 30 min after the first CER injection. Two hours after the last CER injection, we evaluated damage to the pancreas by histological methods; serum levels of amylase, lipase, leukotriene (LT)B₄ and prostaglandin (PG)E₂ ; pancreatic expression of COX-2 and 5-LOX and tumour necrosis factor-α (TNF-α) gene expression by real-time polymerase chain reaction. Caerulein induced inflammatory changes in the pancreas and raised values of the other variables measured. In CER-treated animals, but not in those given saline, flavocoxid inhibited COX-2 and 5-LOX expression, reduced serum levels of lipase and amylase and the degree of pancreatic oedema. Treatment with flavocoxid blunted the increased pancreatic TNF-α mRNA expression, serum leukotriene B₄ and prostaglandin E₂ levels, and protected against histological damage in terms of vacuolization and leukocyte infiltration. Our results confirm the key role of both COX-2 and 5-LOX in the inflammatory response to acute pancreatitis. Flavocoxid may provide a potential therapeutic approach to the treatment of patients at high risk of developing this life-threatening condition. © 2010 The Authors. British Journal of Pharmacology © 2010 The British Pharmacological Society.

CRISPR/Cas9-Mediated Insertion of loxP Sites in the Mouse Dock7 Gene Provides an Effective Alternative to Use of Targeted Embryonic Stem Cells.

PubMed

Bishop, Kathleen A; Harrington, Anne; Kouranova, Evguenia; Weinstein, Edward J; Rosen, Clifford J; Cui, Xiaoxia; Liaw, Lucy

2016-07-07

Targeted gene mutation in the mouse is a primary strategy to understand gene function and relation to phenotype. The Knockout Mouse Project (KOMP) had an initial goal to develop a public resource of mouse embryonic stem (ES) cell clones that carry null mutations in all genes. Indeed, many useful novel mouse models have been generated from publically accessible targeted mouse ES cell lines. However, there are limitations, including incorrect targeting or cassette structure, and difficulties with germline transmission of the allele from chimeric mice. In our experience, using a small sample of targeted ES cell clones, we were successful ∼50% of the time in generating germline transmission of a correctly targeted allele. With the advent of CRISPR/Cas9 as a mouse genome modification tool, we assessed the efficiency of creating a conditional targeted allele in one gene, dedicator of cytokinesis 7 (Dock7), for which we were unsuccessful in generating a null allele using a KOMP targeted ES cell clone. The strategy was to insert loxP sites to flank either exons 3 and 4, or exons 3 through 7. By coinjecting Cas9 mRNA, validated sgRNAs, and oligonucleotide donors into fertilized eggs from C57BL/6J mice, we obtained a variety of alleles, including mice homozygous for the null alleles mediated by nonhomologous end joining, alleles with one of the two desired loxP sites, and correctly targeted alleles with both loxP sites. We also found frequent mutations in the inserted loxP sequence, which is partly attributable to the heterogeneity in the original oligonucleotide preparation. Copyright © 2016 Bishop et al.

The sunburn response in human skin is characterized by sequential eicosanoid profiles that may mediate its early and late phases

PubMed Central

Rhodes, Lesley E.; Gledhill, Karl; Masoodi, Mojgan; Haylett, Ann K.; Brownrigg, Margaret; Thody, Anthony J.; Tobin, Desmond J.; Nicolaou, Anna

2009-01-01

Sunburn is a commonly occurring acute inflammatory process, with dermal vasodilatation and leukocyte infiltration as central features. Ultraviolet (UV) B-induced hydrolysis of membrane phospholipids releases polyunsaturated fatty acids, and their subsequent metabolism by cyclooxygenases (COXs) and lipoxygenases (LOXs) may produce potent eicosanoid mediators modulating different stages of the inflammation. Our objective was to identify candidate eicosanoids formed during the sunburn reaction in relation to its clinical and histological course. We exposed skin of healthy humans (n=32) to UVB and, for 72 h, examined expression of proinflammatory and anti-inflammatory eicosanoids using LC/ESI-MS/MS, and examined immunohistochemical expression of COX-2, 12-LOX, 15-LOX, and leukocyte markers, while quantifying clinical erythema. We show that vasodilatory prostaglandins (PGs) PGE2, PGF2α, and PGE3 accompany the erythema in the first 24–48 h, associated with increased COX-2 expression at 24 h. Novel, potent leukocyte chemoattractants 11-, 12-, and 8-monohydroxy-eicosatetraenoic acid (HETE) are elevated from 4 to 72 h, in association with peak dermal neutrophil influx at 24 h, and increased dermal CD3+ lymphocytes and 12- and 15-LOX expression from 24 to 72 h. Anti-inflammatory metabolite 15-HETE shows later expression, peaking at 72 h. Sunburn is characterized by overlapping sequential profiles of increases in COX products followed by LOX products that may regulate subsequent events and ultimately its resolution.—Rhodes, L. E., Gledhill, K., Masoodi, M., Haylett, A. K., Brownrigg, M., Thody, A. J., Tobin, D. J., Nicolaou, A. The sunburn response in human skin is characterized by sequential eicosanoid profiles that may mediate its early and late phases. PMID:19584301

Development of amperometric lysine biosensors based on Au nanoparticles/multiwalled carbon nanotubes/polymers modified Au electrodes.

PubMed

Chauhan, Nidhi; Singh, Anamika; Narang, Jagriti; Dahiya, Swati; Pundir, C S

2012-11-07

The construction of two amperometric l-lysine biosensors is described in this study. The construction comprises the covalent immobilization of lysine oxidase (LOx) onto nanocomposite composed of gold nanoparticles (AuNPs) and carboxylated multiwalled carbon nanotubes (c-MWCNT), decorated on (i) polyaniline (PANI) and (ii) poly 1,2 diaminobenzene (DAB), electrodeposited on Au electrodes. The biosensors were characterized by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) and electrochemical impedance spectroscopy (EIS) studies. The optimum response (current) was observed within 2 s at pH 7.0 and 25 °C for LOx/AuNPs/c-MWCNT/PANI/Au, and 4 s at pH 7.0 and 30 °C for LOx/AuNPs/c-MWCNT/DAB/Au electrodes. There was a linear relationship between current and lysine concentration ranging from 5.0 to 600 μM for LOx/AuNPs/c-MWCNT/PANI/Au with a detection limit of 5.0 μM, and 20 to 600 μM for the LOx/AuNPs/c-MWCNT/DAB/Au electrode with a detection limit of 20 μM. The PANI modified electrode was in good agreement with the standard HPLC method, with a better correlation (r = 0.992) compared to the DAB modified electrode (r = 0.986). These observations revealed that the PANI modified Au electrode was better than the DAB modified electrode, and hence it was employed for the determination of lysine in milk, pharmaceutical tablets and sera. The PANI modified electrode showed a half life of 120 days, compared to that of 90 days for the DAB modified electrode, after their 100 uses, when stored at 4 °C.

Global Response of the Space Shuttle External Tank with the Presence of Intertank Stringer Cracks and Radius Blocks

NASA Technical Reports Server (NTRS)

Lovejoy, Andrew E.; Rankin, Charles C.

2013-01-01

After propellant was loaded into the external tank (ET), the November 5, 2010 launch of Space Shuttle mission STS-133 was scrubbed due to a gaseous hydrogen leak located in a vent line near the ground umbilical and ET connection. Subsequent visual inspections identified cracks in the sprayed-on foam insulation in the forward end of the ET intertank segment, adjacent to the liquid oxygen (LOX) tank, as shown in Figure 1. These cracks necessitated repair of the foam due to debris concerns that violated launch constraints. As part of the repair process, the affected foam was removed to reveal cracks in the underlying external hat stiffeners on the intertank, as shown in Figure 2. Ultimately, five stiffeners were discovered to be cracked adjacent to the LOX tank. As the managing center for the ET Project, NASA Marshall Space Flight Center (MSFC) coordinated failure investigation and repair activities among multiple organizations, which included the ET prime contractor (Lockheed Martin Space Systems Michoud Operations), the Space Shuttle Program Office at the NASA Johnson Space Center (JSC), the NASA Kennedy Space Center (KSC), and the NASA Engineering and Safety Center (NESC). STS-133 utilized the external tank designated as ET-137. Many aspects of the investigation have been reported previously in Refs. 1-7, which focus on the root cause of the failures, the flight readiness rationale and the local analyses of the stringer failures and repair. This paper summarizes the global analyses that were conducted on ET-137 as part of the NESC effort during the investigation, which was conducted primarily to determine if the repairs that were introduced to the stringers would alter the global response of the ET. In the process of the investigation, a new STAGS tabular input capability was developed to more easily introduce the aerodynamic pressure loads using a method that could easily be extended to incorporate finite element property data such as skin and stiffener thicknesses and beam cross-sectional properties.

Impact sensitivity of materials in contact with liquid and gaseous oxygen at high pressure

NASA Technical Reports Server (NTRS)

Schwinghamer, R. J.

1972-01-01

As a result of the Apollo 13 incident, increased emphasis is being placed on materials compatibility in a high pressure GOX environment. It is known that in addition to impact sensitivity of materials, approximately adiabatic compression conditions can contrive to induce materials reactivity. Test runs at high pressure using the ABMA tester indicate the following: (1) The materials used in the tests showed an inverse relationship between thickness and impact sensitivity. (2) Several materials tested exhibited greater impact sensitivity in GOX than in LOX. (3) The impact sensitivity of the materials tested in GOX, at the pressures tested, showed enhanced impact sensitivity with higher pressure. (4) The rank ordering of the materials tested in LOX up to 1000 psia is the same as the rank ordering resulting from tests in LOX at 14.7 psia.

The benefits of in-flight LOX collection for airbreathing space boosters

NASA Astrophysics Data System (ADS)

Maurice, Lourdes Q.; Leingang, John L.; Carreiro, Louis R.

1992-12-01

In-flight LOX collection using a propulsion fluid system known as ACES (Air Collection and Enrichment System) yields large reductions in launch weights of airbreathing space boosters. The role of the ACES system is to acquire and store liquid oxygen en route to orbit for rocket use beyond the airbreathing envelope. Earth-to-orbit capability is achieved without carrying liquid oxygen from take-off or relying on scramjets. The superiority of ACES type space boosters over their LOX-carrying counterparts has been thoroughly documented in the past. This paper extends that work by presenting a direct comparison between single-stage and two-stage ACES and scramjet powered vehicles carrying similar payloads. ACES vehicles are shown to be weight competitive with scramjet powered vehicles, and require airbreathing function only up to Mach 5 to 8.

NASA X-34 Technology in Motion

NASA Technical Reports Server (NTRS)

Beech, Geoffrey; Chandler, Kristie

1997-01-01

The X-34 technology development program is a joint industry/government project to develop, test, and operate a small, fully-reusable hypersonic flight vehicle. The objective is to demonstrate key technologies and operating concepts applicable to future reusable launch vehicles. Integrated in the vehicle are various systems to assure successful completion of mission objectives, including the Main Propulsion System (MPS). NASA-Marshall Space Flight Center (MSFC) is responsible for developing the X-34's MPS including the design and complete build package for the propulsion system components. The X-34 will be powered by the Fastrac Engine, which is currently in design and development at NASA-MSFC. Fastrac is a single-stage main engine, which burns a mixture of liquid oxygen (LOX) and kerosene(RP-1). The interface between the MPS and Fastrac engine are critical for proper system operation and technologies applicable to future reusable launch vehicles. Deneb's IGRIP software package with the Dynamic analysis option provided a key tool for conducting studies critical to this interface as well as a mechanism to drive the design of the LOX and RP-1 feedlines. Kinematic models were created for the Fastrac Engine and the feedlines for various design concepts. Based on the kinematic simulation within Envision, design and joint limits were verified and system interference controlled. It was also critical to the program to evaluate the effect of dynamic loads visually, providing a verification tool for dynamic analysis and in some cases uncovering areas that had not been considered. Deneb's software put the X-34 technology in motion and has been a key factor in facilitating the strenuous design schedule.

Fosmid Cre-LoxP Inverse PCR Paired-End (Fosmid CLIP-PE), a Novel Method for Constructing Fosmid Pair-End Library (Seventh Annual Sequencing, Finishing, Analysis in the Future (SFAF) Meeting 2012)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Peng, Ze

2012-06-01

Ze Peng from DOE JGI presents "Fosmid Cre-LoxP Inverse PCR Paired-End (Fosmid CLIP-PE), a Novel Method for Constructing Fosmid Pair-End Library" at the 7th Annual Sequencing, Finishing, Analysis in the Future (SFAF) Meeting held in June, 2012 in Santa Fe, NM.

Simulation and Application of GPOPS for a Trajectory Optimization and Mission Planning Tool

DTIC Science & Technology

2010-03-01

12,000lbf) vaccum Specific Impulse 269 s 455 s 316 s Burn Time 124 s 480 s 1250s Fuel Solid LOX/ LH2 MMH/N2O4 Height 184 ft Diameter 28.5 ft...285,000 lb Engine 2 J-2S Linear Aerospikes Thrust 410,000 lbf Fuel LOX/ LH2 20 Figure 9: Minuteman Launch [29] Currently the main missile

Fosmid Cre-LoxP Inverse PCR Paired-End (Fosmid CLIP-PE), a Novel Method for Constructing Fosmid Pair-End Library (Seventh Annual Sequencing, Finishing, Analysis in the Future (SFAF) Meeting 2012)

ScienceCinema

Peng, Ze

2018-01-24

Ze Peng from DOE JGI presents "Fosmid Cre-LoxP Inverse PCR Paired-End (Fosmid CLIP-PE), a Novel Method for Constructing Fosmid Pair-End Library" at the 7th Annual Sequencing, Finishing, Analysis in the Future (SFAF) Meeting held in June, 2012 in Santa Fe, NM.

CRISPR/Cas9-loxP-Mediated Gene Editing as a Novel Site-Specific Genetic Manipulation Tool.

PubMed

Yang, Fayu; Liu, Changbao; Chen, Ding; Tu, Mengjun; Xie, Haihua; Sun, Huihui; Ge, Xianglian; Tang, Lianchao; Li, Jin; Zheng, Jiayong; Song, Zongming; Qu, Jia; Gu, Feng

2017-06-16

Cre-loxP, as one of the site-specific genetic manipulation tools, offers a method to study the spatial and temporal regulation of gene expression/inactivation in order to decipher gene function. CRISPR/Cas9-mediated targeted genome engineering technologies are sparking a new revolution in biological research. Whether the traditional site-specific genetic manipulation tool and CRISPR/Cas9 could be combined to create a novel genetic tool for highly specific gene editing is not clear. Here, we successfully generated a CRISPR/Cas9-loxP system to perform gene editing in human cells, providing the proof of principle that these two technologies can be used together for the first time. We also showed that distinct non-homologous end-joining (NHEJ) patterns from CRISPR/Cas9-mediated gene editing of the targeting sequence locates at the level of plasmids (episomal) and chromosomes. Specially, the CRISPR/Cas9-mediated NHEJ pattern in the nuclear genome favors deletions (64%-68% at the human AAVS1 locus versus 4%-28% plasmid DNA). CRISPR/Cas9-loxP, a novel site-specific genetic manipulation tool, offers a platform for the dissection of gene function and molecular insights into DNA-repair pathways. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

Impact of wines and wine constituents on cyclooxygenase-1, cyclooxygenase-2, and 5-lipoxygenase catalytic activity.

PubMed

Kutil, Zsofia; Temml, Veronika; Maghradze, David; Pribylova, Marie; Dvorakova, Marcela; Schuster, Daniela; Vanek, Tomas; Landa, Premysl

2014-01-01

Cyclooxygenases and lipoxygenases are proinflammatory enzymes; the former affects platelet aggregation, vasoconstriction, vasodilatation and later the development of atherosclerosis. Red wines from Georgia and central and western Europe inhibited cyclooxygenase-1 (COX-1) activity in the range of 63-94%, cyclooxygenase-2 (COX-2) activity in the range of 20-44% (tested at a concentration of 5 mL/L), and 5-lipoxygenase (5-LOX) activity in the range of 72-84% (at a concentration of 18.87 mL/L). White wines inhibited 5-LOX in the range of 41-68% at a concentration of 18.87 mL/L and did not inhibit COX-1 and COX-2. Piceatannol (IC50 = 0.76 μM) was identified as a strong inhibitor of 5-LOX followed by luteolin (IC50 = 2.25 μM), quercetin (IC50 = 3.29 μM), and myricetin (IC50 = 4.02 μM). trans-Resveratrol was identified as an inhibitor of COX-1 (IC50 = 2.27 μM) and COX-2 (IC50 = 3.40 μM). Red wine as a complex mixture is a powerful inhibitor of COX-1, COX-2, and 5-LOX, the enzymes involved in eicosanoid biosynthetic pathway.

Multitarget molecular hybrids of cinnamic acids.

PubMed

Peperidou, Aikaterini; Kapoukranidou, Dorothea; Kontogiorgis, Christos; Hadjipavlou-Litina, Dimitra

2014-12-02

In an attempt to synthesize potential new multitarget agents, 11 novel hybrids incorporating cinnamic acids and paracetamol, 4-/7-hydroxycoumarin, benzocaine, p-aminophenol and m-aminophenol were synthesized. Three hybrids-2e, 2a, 2g-and 3b were found to be multifunctional agents. The hybrid 2e derived from the phenoxyphenyl cinnamic acid and m-acetamidophenol showed the highest lipoxygenase (LOX) inhibition and analgesic activity (IC50 = 0.34 μΜ and 98.1%, whereas the hybrid 3b of bromobenzyloxycinnamic acid and hymechromone exhibited simultaneously good LOX inhibitory activity (IC50 = 50 μΜ) and the highest anti-proteolytic activity (IC50= 5 μΜ). The hybrid 2a of phenyloxyphenyl acid with paracetamol showed a high analgesic activity (91%) and appears to be a promising agent for treating peripheral nerve injuries. Hybrid 2g which has an ester and an amide bond presents an interesting combination of anti-LOX and anti-proteolytic activity. The esters were found very potent and especially those derived from paracetamol and m-acetamidophenol. The amides follow. Based on 2D-structure-activity relationships it was observed that both steric and electronic parameters play major roles in the activity of these compounds. Molecular docking studies point to the fact that allosteric interactions might govern the LOX-inhibitor binding.

High-salt in addition to high-fat diet may enhance inflammation and fibrosis in liver steatosis induced by oxidative stress and dyslipidemia in mice.

PubMed

Uetake, Yuzaburo; Ikeda, Hitoshi; Irie, Rie; Tejima, Kazuaki; Matsui, Hiromitsu; Ogura, Sayoko; Wang, Hong; Mu, ShengYu; Hirohama, Daigoro; Ando, Katsuyuki; Sawamura, Tatsuya; Yatomi, Yutaka; Fujita, Toshiro; Shimosawa, Tatsuo

2015-02-13

It is widely known that salt is an accelerating factor for the progression of metabolic syndrome and causes cardiovascular diseases, most likely due to its pro-oxidant properties. We hypothesized that excessive salt intake also facilitates the development of nonalcoholic steatohepatitis (NASH), which is frequently associated with metabolic syndrome. We examined the exacerbating effect of high-salt diet on high-fat diet-induced liver injury in a susceptible model to oxidative stress, apoE knockout and lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) transgenic mice. High-salt diet led to NASH in high-fat diet-fed LOX-1 transgenic/apoE knockout mice without affecting high-fat diet-induced dyslipidemia or hepatic triglyceride accumulation. Additionally, a high-salt and high-fat diet stimulated oxidative stress production and inflammatory reaction to a greater extent than did a high-fat diet in the liver of LOX-1 transgenic/apoE knockout mice. We demonstrated that high-salt diet exacerbated NASH in high-fat diet-fed LOX-1 transgenic /apoE knockout mice and that this effect was associated with the stimulation of oxidative and inflammatory processes; this is the first study to suggest the important role of excessive salt intake in the development of NASH.

Development tests of LOX/LH 2 tank for H-I launch vehicle

NASA Astrophysics Data System (ADS)

Takamatsu, H.; Imagawa, K.; Ichimaru, Y.

H-I is a future launch vehicle of Japan with a capability of placing more than 550 kg payload into a geostationary orbit. The National Space Development Agency of Japan (NASDA) is now directing its efforts to the final development of H-I launch vehicle. H-I's high launch capability is attained by adopting a newly developed second stage with a LOX/LH 2 propulsion system. The second stage propulsion system consists of a tank and an engine. The tank is 2.5 m in diameter and 5.7 m in length and contains 8.7 tons of propellants. This tank is an integral tank with a common bulkhead which separates the tank into forward LH 2 tank and aft LOX tank. The tank is made of 2219 aluminum alloy and is insulated with sprayed polyurethane foam. The common bulkhead is made of FRP honeycomb core and aluminium alloy surface sheets. The most critical item in the development of the tank is the common bulkhead, therefore the cryogenic structural test was carried out to verify the structural integrity of the bulkhead. The structural integrity of the whole LOX/LH 2 tank was verified by the cryogenic structural test of a sub-scale tank and the room temperature structural test of a prototype tank.

Lipoxygenase6-Dependent Oxylipin Synthesis in Roots Is Required for Abiotic and Biotic Stress Resistance of Arabidopsis1[C][W

PubMed Central

Grebner, Wiebke; Stingl, Nadja E.; Oenel, Ayla; Mueller, Martin J.; Berger, Susanne

2013-01-01

Jasmonates are oxylipin signals that play important roles in the development of fertile flowers and in defense against pathogens and herbivores in leaves. The aim of this work was to understand the synthesis and function of jasmonates in roots. Grafting experiments with a jasmonate-deficient mutant demonstrated that roots produce jasmonates independently of leaves, despite low expression of biosynthetic enzymes. Levels of 12-oxo-phytodienoic acid, jasmonic acid, and its isoleucine derivative increased in roots upon osmotic and drought stress. Wounding resulted in a decrease of preformed 12-oxo-phytodienoic acid concomitant with an increase of jasmonic acid and jasmonoyl-isoleucine. 13-Lipoxygenases catalyze the first step of lipid oxidation leading to jasmonate production. Analysis of 13-lipoxygenase-deficient mutant lines showed that only one of the four 13-lipoxygenases, LOX6, is responsible and essential for stress-induced jasmonate accumulation in roots. In addition, LOX6 was required for production of basal 12-oxo-phytodienoic acid in leaves and roots. Loss-of-function mutants of LOX6 were more attractive to a detritivorous crustacean and more sensitive to drought, indicating that LOX6-derived oxylipins are important for the responses to abiotic and biotic factors. PMID:23444343

Impact of Wines and Wine Constituents on Cyclooxygenase-1, Cyclooxygenase-2, and 5-Lipoxygenase Catalytic Activity

PubMed Central

Temml, Veronika; Maghradze, David; Vanek, Tomas

2014-01-01

Cyclooxygenases and lipoxygenases are proinflammatory enzymes; the former affects platelet aggregation, vasoconstriction, vasodilatation and later the development of atherosclerosis. Red wines from Georgia and central and western Europe inhibited cyclooxygenase-1 (COX-1) activity in the range of 63–94%, cyclooxygenase-2 (COX-2) activity in the range of 20–44% (tested at a concentration of 5 mL/L), and 5-lipoxygenase (5-LOX) activity in the range of 72–84% (at a concentration of 18.87 mL/L). White wines inhibited 5-LOX in the range of 41–68% at a concentration of 18.87 mL/L and did not inhibit COX-1 and COX-2. Piceatannol (IC50 = 0.76 μM) was identified as a strong inhibitor of 5-LOX followed by luteolin (IC50 = 2.25 μM), quercetin (IC50 = 3.29 μM), and myricetin (IC50 = 4.02 μM). trans-Resveratrol was identified as an inhibitor of COX-1 (IC50 = 2.27 μM) and COX-2 (IC50 = 3.40 μM). Red wine as a complex mixture is a powerful inhibitor of COX-1, COX-2, and 5-LOX, the enzymes involved in eicosanoid biosynthetic pathway. PMID:24976682

Up-regulation of 5-lipoxygenase by inhibition of cathepsin G enhances TRAIL-induced apoptosis through down-regulation of survivin

PubMed Central

Woo, Seon Min; Min, Kyoung-Jin; Seo, Seung Un; Kim, Shin; Park, Jong-Wook; Song, Dae Kyu; Lee, Hyun-Shik; Kim, Sang Hyun; Kwon, Taeg Kyu

2017-01-01

Cathepsin G is a serine protease secreted from activated neutrophils, it has important roles in inflammation and immune response. Moreover, cathepsin G promotes tumor cell-cell adhesion and migration in cancer cells. In this study, we investigated whether inhibition of cathepsin G could sensitize TRAIL-mediated apoptosis in cancer cells. An inhibitor of cathepsin G [Cathepsin G inhibitor I (Cat GI); CAS 429676-93-7] markedly induced TRAIL-mediated apoptosis in human renal carcinoma (Caki, ACHN, and A498), lung cancer (A549) and cervical cancer (Hela) cells. In contrast, combined treatment with Cat GI and TRAIL had no effect on apoptosis in normal cells [mesangial cell (MC) and human skin fibroblast (HSF)]. Cat GI induced down-regulation of survivin expression at the post-translational level, and overexpression of survivin markedly blocked apoptosis induced by combined treatment with Cat GI plus TRAIL. Interestingly, Cat GI induced down-regulation of survivin via 5-lipoxygenase (5-LOX)-mediated reactive oxygen species (ROS) production. Inhibition of 5-LOX by gene silencing (siRNA) or a pharmacological inhibitor of 5-LOX (zileuton) markedly attenuated combined treatment-induced apoptosis. Taken together, our results indicate that inhibition of cathepsin G sensitizes TRAIL-induced apoptosis through 5-LOX-mediated down-regulation of survivin expression. PMID:29290980

Lipoxygenase in Caragana jubata responds to low temperature, abscisic acid, methyl jasmonate and salicylic acid.

PubMed

Bhardwaj, Pardeep Kumar; Kaur, Jagdeep; Sobti, Ranbir Chander; Ahuja, Paramvir Singh; Kumar, Sanjay

2011-09-01

Lipoxygenase (LOX) catalyses oxygenation of free polyunsaturated fatty acids into oxylipins, and is a critical enzyme of the jasmonate signaling pathway. LOX has been shown to be associated with biotic and abiotic stress responses in diverse plant species, though limited data is available with respect to low temperature and the associated cues. Using rapid amplification of cDNA ends, a full-length cDNA (CjLOX) encoding lipoxygenase was cloned from apical buds of Caragana jubata, a temperate plant species that grows under extreme cold. The cDNA obtained was 2952bp long consisting of an open reading frame of 2610bp encoding 869 amino acids protein. Multiple alignment of the deduced amino acid sequence with those of other plants demonstrated putative LH2/ PLAT domain, lipoxygenase iron binding catalytic domain and lipoxygenase_2 signature sequences. CjLOX exhibited up- and down-regulation of gene expression pattern in response to low temperature (LT), abscisic acid (ABA), methyl jasmonate (MJ) and salicylic acid (SA). Among all the treatments, a strong up-regulation was observed in response to MJ. Data suggests an important role of jasmonate signaling pathway in response to LT in C. jubata. Copyright © 2011 Elsevier B.V. All rights reserved.

Long Duration Exposure Facility (LDEF) structural verification test report

NASA Technical Reports Server (NTRS)

Jones, T. C.; Lucy, M. H.; Shearer, R. L.

1983-01-01

Structural load tests on the Long Duration Exposure Facility's (LDEF) primary structure were conducted. These tests had three purposes: (1) demonstrate structural adequacy of the assembled LDEF primary structure when subjected to anticipated flight loads; (2) verify analytical models and methods used in loads and stress analysis; and (3) perform tests to comply with the Space Transportation System (STS) requirements. Test loads were based on predicted limit loads which consider all flight events. Good agreement is shown between predicted and observed load, strain, and deflection data. Test data show that the LDEF structure was subjected to 1.2 times limit load to meet the STS requirements. The structural adequacy of the LDEF is demonstrated.

Characterization of a Pressure-Fed LOX/LCH4 Reaction Control System Under Simulated Altitude and Thermal Vacuum Conditions

NASA Technical Reports Server (NTRS)

Atwell, Matthew J.; Melcher, John C.; Hurlbert, Eric A.; Morehead, Robert L.

2017-01-01

A liquid oxygen, liquid methane (LOX/LCH4) reaction control system (RCS) was tested at NASA Glenn Research Center's Plum Brook Station in the Spacecraft Propulsion Research Facility (B-2) under simulated altitude and thermal vacuum conditions. The RCS is a subsystem of the Integrated Cryogenic Propulsion Test Article (ICPTA) and was initially developed under Project Morpheus. Composed of two 28 lbf-thrust and two 7 lbf-thrust engines, the RCS is fed in parallel with the ICPTA main engine from four propellant tanks. 40 tests consisting of 1,010 individual thruster pulses were performed across 6 different test days. Major test objectives were focused on system dynamics, and included characterization of fluid transients, manifold priming, manifold thermal conditioning, thermodynamic vent system (TVS) performance, and main engine/RCS interaction. Peak surge pressures from valve opening and closing events were examined. It was determined that these events were impacted significantly by vapor cavity formation and collapse. In most cases the valve opening transient was more severe than the valve closing. Under thermal vacuum conditions it was shown that TVS operation is unnecessary to maintain liquid conditions at the thruster inlets. However, under higher heat leak environments the RCS can still be operated in a self-conditioning mode without overboard TVS venting, contingent upon the engines managing a range of potentially severe thermal transients. Lastly, during testing under cold thermal conditions the engines experienced significant ignition problems. Only after warming the thruster bodies with a gaseous nitrogen purge to an intermediate temperature was successful ignition demonstrated.

Aircraft Landing Dynamics Facility - A unique facility with new capabilities

NASA Technical Reports Server (NTRS)

Davis, P. A.; Stubbs, S. M.; Tanner, J. A.

1985-01-01

The Aircraft Landing Dynamics Facility (ALDF), formerly called the Landing Loads Track, is described. The paper gives a historical overview of the original NASA Langley Research Center Landing Loads Track and discusses the unique features of this national test facility. Comparisons are made between the original track characteristics and the new capabilities of the Aircraft Landing Dynamics Facility following the recently completed facility update. Details of the new propulsion and arresting gear systems are presented along with the novel features of the new high-speed carriage. The data acquisition system is described and the paper concludes with a review of future test programs.

Raymond International Inc. will construct twin berthing facilities for loading and offloading crude

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

1978-03-06

Raymond International Inc. will construct twin berthing facilities for loading and offloading crude in St. James Parish, La. The $16.6 million contract was let under the Strategic Petroleum Reserve program. Completion is expected in 1978.

Synthetic Minor NSR Permit: Thunder Butte Petroleum Services, Inc. - Crude Storage and Loading Facility

EPA Pesticide Factsheets

This page contains documents relevant to the synthetic minor NSR permi for the Thunder Butte Petroleum Services, Inc., Crude Storage and Loading Facility, located on the Fort Berthold Indian Reservation in Ward County, ND.

One Idea for a Next Generation Shuttle

NASA Technical Reports Server (NTRS)

MacConochie, Ian O.; Cerro, Jeffrey A.

2004-01-01

In this configuration, the current Shuttle External Tank serves as core structure for a fully reusable second stage. This stage is equipped with wings, vertical fin, landing gear, and thermal protection. The stage is geometrically identical to (but smaller than) a single stage that has been tested hyper-sonically, super-sonically, and sub-sonically in the NASA Langley Research Center wind tunnels. The three LOX/LH engines that currently serve as main propulsion for the Shuttle Orbiter, serve as main propulsion on the new stage. The new stage is unmanned but is equipped with the avionics needed for automatic maneuvering on orbit and for landing on a runway. Three rails are installed along the top surface of the vehicle for attachment of various payloads. Pay- loads might include third stages with satellites attached, personnel pods, propellants, or other items.

Impact of lunar oxygen production on direct manned Mars missions

NASA Technical Reports Server (NTRS)

Young, Roy M., Jr.; Tucker, William B.

1992-01-01

A manned Mars program made up of six missions is evaluated to determine the impact of using lunar liquid oxygen (LOX) as a propellant. Two departure and return nodes, low Earth orbit and low lunar orbit, are considered, as well as two return vehicle configurations, a full 70,000-kg vehicle and a 6800-kg capsule. The cost of lunar LOX delivered to orbit is expressed as a ratio of Earth launch cost.

In Vivo Imaging of MDR1A Gene Expression

DTIC Science & Technology

2004-12-01

Engineer PGK-neo and Renilla luciferase cassettes, already available, with appropriate loxP sites, into mdrla locus. Repeat for HSV-tk reporter. The...of the gene-targeting vector. under the control of the PGK promoter. Luc: Renilla luciferase fused in- frame with the translated sequences of exon 2...between the two loxP sites, upstream of the Neo cassette. A cloning strategy was then devised to fuse Renilla luciferase in-frame with the translated

Axisymmetric single shear element combustion instability experiment

NASA Technical Reports Server (NTRS)

Breisacher, Kevin J.

1993-01-01

The combustion stability characteristics of a combustor consisting of a single shear element and a cylindrical chamber utilizing LOX and gaseous hydrogen as propellants are presented. The combustor geometry and the resulting longitudinal mode instability are axisymmetric. Hydrogen injection temperature and pyrotechnic pulsing were used to determine stability boundaries. Mixture ratio, fuel annulus gap, and LOX post configuration were varied. Performance and stability data were obtained for chamber pressures of 300 and 1000 psia.

Axisymmetric single shear element combustion instability experiment

NASA Technical Reports Server (NTRS)

Breisacher, Kevin J.

1993-01-01

The combustion stability characteristics of a combustor consisting of a single shear element and a cylindrical chamber utilizing LOX and gaseous hydrogen as propellants are presented. The combustor geometry and the resulting longitudinal mode instability are axisymmetric. Hydrogen injection temperature and pyrotechnic pulsing were used to determine stability boundaries. Mixture ratio, fuel annulus gap, and LOX post configuration were varied. Performance and stability data are presented for chamber pressures of 300 and 1000 psia.

Non-Toxic Orbital Maneuvering System Engine Development

NASA Technical Reports Server (NTRS)

Green, Christopher; Claflin, Scott; Maeding, Chris; Butas, John

1999-01-01

Recent results using the Aestus engine operated with LOx/ethanol propellant are presented. An experimental program at Rocketdyne Propulsion and Power is underway to adapt this engine for the Boeing Reusable Space Systems Division non-toxic Orbital Maneuvering System/Reaction control System (OMS/RCS) system. Daimler-Chrysler Aerospace designed the Aestus as an nitrogen tetroxide/monomethyl hydrazine (NTO/MMH) upper-stage engine for the Ariane 5. The non-toxic OMS/RCS system's preliminary design requires a LOx/ethanol (O2/C2H5OH) engine that operates with a mixture ratio of 1.8, a specific impulse of 323 seconds, and fits within the original OMS design envelope. This paper describes current efforts to meet these requirements including, investigating engine performance using LOx/ethanol, developing the en-ine system sizing package, and meeting the vehicle operation parameters. Data from hot-fire testing are also presented and discussed.

POGO analysis based on N-II/H-I vehicle flight data

NASA Astrophysics Data System (ADS)

Mori, Hidehiko

Three types of longitudinal oscillations Pre-MECO POGO 1, Pre-MECO POGO 2, and MECO POGO have been observed in the launches of N-II/H-I vehicles. A Nyquist plot of a mathematical POGO model is used to examine stability properties of these oscillations. Pre-MECO POGO 1 and MECO POGO are generated in the LOX feed system installed with a accumulator. Flow fluctuation due to the LOX pump vibration is the main exciting factor for the former, the fluctuation of LOX tank bottom pressure for the latter. Pre-MECO POGO 2, excited in the vicinity of open-pipe resonant frequency of fuel suction line, is affected by fuel flow fluctuation. Frequency, longitudinal structural mode shape, and generalized mass related to each POGO are determined from flight data. The POGO model with these parameters is shown to represent the whole POGO features of N-II/H-I along flight time.

Mammalian lipoxygenases and their biological relevance

PubMed Central

Kuhn, Hartmut; Banthiya, Swathi; van Leyen, Klaus

2015-01-01

Lipoxygenases (LOXs) form a heterogeneous class of lipid peroxidizing enzymes, which have been implicated in cell proliferation and differentiation but also in the pathogenesis of various diseases with major public health relevance. As other fatty acid dioxygenases LOX oxidize polyunsaturated fatty acids to their corresponding hydroperoxy derivatives, which are further transformed to bioactive lipid mediators (eicosanoids and related substances). On the other hand, lipoxygenases are key players in regulation of the cellular redox homeostasis, which is an important element in gene expression regulation. Although the first mammalian lipoxygenases were discovered 40 years ago and although the enzymes have been well characterized with respect to their structural and functional properties the biological roles of the different lipoxygenase isoforms are not completely understood. This review is aimed at summarizing the current knowledge on the physiological roles of different mammalian LOX-isoforms and their patho-physiological function in inflammatory, metabolic, hyperproliferative, neurodegenerative and infectious disorders. PMID:25316652

Liquid Acquisition Device Testing with Sub-Cooled Liquid Oxygen

NASA Technical Reports Server (NTRS)

Jurns, John M.; McQuillen, John B.

2008-01-01

When transferring propellant in space, it is most efficient to transfer single phase liquid from a propellant tank to an engine. In earth s gravity field or under acceleration, propellant transfer is fairly simple. However, in low gravity, withdrawing single-phase fluid becomes a challenge. A variety of propellant management devices (PMD) are used to ensure single-phase flow. One type of PMD, a liquid acquisition device (LAD) takes advantage of capillary flow and surface tension to acquire liquid. Previous experimental test programs conducted at NASA have collected LAD data for a number of cryogenic fluids, including: liquid nitrogen (LN2), liquid oxygen (LOX), liquid hydrogen (LH2), and liquid methane (LCH4). The present work reports on additional testing with sub-cooled LOX as part of NASA s continuing cryogenic LAD development program. Test results extend the range of LOX fluid conditions examined, and provide insight into factors affecting predicting LAD bubble point pressures.

Effects of Compounded Stanford Modified Oral Rinse (MucoLox) on the Survival and Migration of Oral Keratinocytes and Fibroblasts: Implications for Wound Healing.

PubMed

Song, Guiyun; Banov, Daniel; Bassani, August S

2018-01-01

Several oral rinses are commercially available to alleviate the symptoms of oral mucositis. Prolonged retention of active pharmaceutical ingredients in the oral cavity is a major problem. In this study, we modified the Stanford oral rinse by including a proprietary mucoadhesive polymer called MucoLox, which we hypothesized would improve active pharmaceutical ingredient mucoadhesion. Characterization of this newly compounded oral rinse showed absence of cytotoxicity in human oral keratinocyte and fibroblast cell lines. The compounded formulation significantly stimulated the migration of these two cell lines in Oris Cell Migration Assay plates, better than the reference commercial product Magic mouthwash. Based on this in vitro study, the new Stanford modified oral rinse with MucoLox is safe and may promote healing of oral mucositis. Copyright© by International Journal of Pharmaceutical Compounding, Inc.

Damage Tolerance Analysis of a Pressurized Liquid Oxygen Tank

NASA Technical Reports Server (NTRS)

Forth, Scott C.; Harvin, Stephen F.; Gregory, Peyton B.; Mason, Brian H.; Thompson, Joe E.; Hoffman, Eric K.

2006-01-01

A damage tolerance assessment was conducted of an 8,000 gallon pressurized Liquid Oxygen (LOX) tank. The LOX tank is constructed of a stainless steel pressure vessel enclosed by a thermal-insulating vacuum jacket. The vessel is pressurized to 2,250 psi with gaseous nitrogen resulting in both thermal and pressure stresses on the tank wall. Finite element analyses were performed on the tank to characterize the stresses from operation. Engineering material data was found from both the construction of the tank and the technical literature. An initial damage state was assumed based on records of a nondestructive inspection performed on the tank. The damage tolerance analyses were conducted using the NASGRO computer code. This paper contains the assumptions, and justifications, made for the input parameters to the damage tolerance analyses and the results of the damage tolerance analyses with a discussion on the operational safety of the LOX tank.

Megakaryocyte pathology and bone marrow fibrosis: the lysyl oxidase connection

PubMed Central

Matsuura, Shinobu

2012-01-01

Megakaryocytes (MKs), the platelet precursors, are capable of accumulating DNA greater than a diploid content as part of their cell cycle. MKs have been recognized as mediating fibrosis in a subset of hematologic malignancies, including acute megakaryoblastic leukemia and a subset of myeloproliferative neoplasms. The mechanisms responsible for fibrosis remain only partially understood. Past studies highlighted the role of growth factors in such pathologies, and recently, the protein lysyl oxidase (LOX) has been implicated in proliferation of MKs, ploidy and deposition of fibers. LOX was initially characterized as a protein responsible for the intermolecular cross-linking of elastin and collagen, and in recent years it has been identified as regulator of various pathologies, such as cancer and inflammation. Here, we review recent advances in the understanding of the contribution of MKs to the progression of myelofibrosis, highlighting the newly identified role of LOX. PMID:22767499

Magnetic Field, Force, and Inductance Computations for an Axially Symmetric Solenoid

NASA Technical Reports Server (NTRS)

Lane, John E.; Youngquist, Robert C.; Immer, Christopher D.; Simpson, James C.

2001-01-01

The pumping of liquid oxygen (LOX) by magnetic fields (B field), using an array of electromagnets, is a current topic of research and development at Kennedy Space Center, FL. Oxygen is paramagnetic so that LOX, like a ferrofluid, can be forced in the direction of a B field gradient. It is well known that liquid oxygen has a sufficient magnetic susceptibility that a strong magnetic gradient can lift it in the earth's gravitational field. It has been proposed that this phenomenon can be utilized in transporting (i.e., pumping) LOX not only on earth, but on Mars and in the weightlessness of space. In order to design and evaluate such a magnetic pumping system, it is essential to compute the magnetic and force fields, as well as inductance, of various types of electromagnets (solenoids). In this application, it is assumed that the solenoids are air wrapped, and that the current is essentially time independent.

Synthetic Minor NSR Permit: Van Hook Crude Terminal, LLC - Van Hook Crude Terminal/Rail Loading Facility

EPA Pesticide Factsheets

This page contains the current effective synthetic minor NSR permit for the Van Hook Crude Terminal, LLC, Van Hook Crude Terminal/Rail Loading Facility, located on the Fort Berthold Indian Reservation in Mountrail County, ND.

Construction comparison of Louisiana's conventional and alternative base courses under accelerated loading : final report.

DOT National Transportation Integrated Search

2001-11-01

This report describes the test results of the first project at the Louisiana Transportation Research Center's Accelerated Loading Facility (ALF). In 1995, 9 test lanes were constructed at the Louisiana Pavement Research Facility in Port Allen. These ...

Uranium-Loaded Water Treatment Resins: 'Equivalent Feed' at NRC and Agreement State-Licensed Uranium Recovery Facilities - 12094

DOE Office of Scientific and Technical Information (OSTI.GOV)

Camper, Larry W.; Michalak, Paul; Cohen, Stephen

Community Water Systems (CWSs) are required to remove uranium from drinking water to meet EPA standards. Similarly, mining operations are required to remove uranium from their dewatering discharges to meet permitted surface water discharge limits. Ion exchange (IX) is the primary treatment strategy used by these operations, which loads uranium onto resin beads. Presently, uranium-loaded resin from CWSs and mining operations can be disposed as a waste product or processed by NRC- or Agreement State-licensed uranium recovery facilities if that licensed facility has applied for and received permission to process 'alternate feed'. The disposal of uranium-loaded resin is costly andmore » the cost to amend a uranium recovery license to accept alternate feed can be a strong disincentive to commercial uranium recovery facilities. In response to this issue, the NRC issued a Regulatory Issue Summary (RIS) to clarify the agency's policy that uranium-loaded resin from CWSs and mining operations can be processed by NRC- or Agreement State-licensed uranium recovery facilities without the need for an alternate feed license amendment when these resins are essentially the same, chemically and physically, to resins that licensed uranium recovery facilities currently use (i.e., equivalent feed). NRC staff is clarifying its current alternate feed policy to declare IX resins as equivalent feed. This clarification is necessary to alleviate a regulatory and financial burden on facilities that filter uranium using IX resin, such as CWSs and mine dewatering operations. Disposing of those resins in a licensed facility could be 40 to 50 percent of the total operations and maintenance (O and M) cost for a CWS. Allowing uranium recovery facilities to treat these resins without requiring a license amendment lowers O and M costs and captures a valuable natural resource. (authors)« less

Morpheus: Advancing Technologies for Human Exploration

NASA Technical Reports Server (NTRS)

Olansen, Jon B.; Munday, Stephen R.; Mitchell, Jennifer D.; Baine, Michael

2012-01-01

NASA's Morpheus Project has developed and tested a prototype planetary lander capable of vertical takeoff and landing. Designed to serve as a vertical testbed (VTB) for advanced spacecraft technologies, the vehicle provides a platform for bringing technologies from the laboratory into an integrated flight system at relatively low cost. This allows individual technologies to mature into capabilities that can be incorporated into human exploration missions. The Morpheus vehicle is propelled by a LOX/Methane engine and sized to carry a payload of 1100 lb to the lunar surface. In addition to VTB vehicles, the Project s major elements include ground support systems and an operations facility. Initial testing will demonstrate technologies used to perform autonomous hazard avoidance and precision landing on a lunar or other planetary surface. The Morpheus vehicle successfully performed a set of integrated vehicle test flights including hot-fire and tethered hover tests, leading up to un-tethered free-flights. The initial phase of this development and testing campaign is being conducted on-site at the Johnson Space Center (JSC), with the first fully integrated vehicle firing its engine less than one year after project initiation. Designed, developed, manufactured and operated in-house by engineers at JSC, the Morpheus Project represents an unprecedented departure from recent NASA programs that traditionally require longer, more expensive development lifecycles and testing at remote, dedicated testing facilities. Morpheus testing includes three major types of integrated tests. A hot-fire (HF) is a static vehicle test of the LOX/Methane propulsion system. Tether tests (TT) have the vehicle suspended above the ground using a crane, which allows testing of the propulsion and integrated Guidance, Navigation, and Control (GN&C) in hovering flight without the risk of a vehicle departure or crash. Morpheus free-flights (FF) test the complete Morpheus system without the additional safeguards provided during tether. A variety of free-flight trajectories are planned to incrementally build up to a fully functional Morpheus lander capable of flying planetary landing trajectories. In FY12, these tests will culminate with autonomous flights simulating a 1 km lunar approach trajectory, hazard avoidance maneuvers and precision landing in a prepared hazard field at the Kennedy Space Center (KSC). This paper describes Morpheus integrated testing campaign, infrastructure, and facilities, and the payloads being incorporated on the vehicle. The Project s fast pace, rapid prototyping, frequent testing, and lessons learned depart from traditional engineering development at JSC. The Morpheus team employs lean, agile development with a guiding belief that technologies offer promise, but capabilities offer solutions, achievable without astronomical costs and timelines.

Vortex shedding experiment with flat and curved bluff plates in water

NASA Technical Reports Server (NTRS)

Reed, D.; Nesman, T.; Howard, P.

1988-01-01

Vortex shedding experiments were conducted in a water flow facility in order to simulate the strong discrete 4000-Hz vibration detected in the Space Shuttle Main Engine (SSME) which is thought to be associated with the SSME LOX inlet tee splitter vanes on the Main Injector. For the case of a flat vane with a blunt trailing edge excited by flow induced vortex shedding, lock-in with the first bending mode of the plate was observed. A curved vane displayed similar behavior, with the lock-in being a more discrete higher amplitude response. Aluminum vanes were employed to decouple the first vane bending mode from the vortex shedding mode. The application of an asymmetric 30-deg trailing edge bevel to both the flat and curved vanes was found to greatly reduce the strength of the shed vortices.

Ice/frost detection using millimeter wave radiometry. [space shuttle external tank

NASA Technical Reports Server (NTRS)

Gagliano, J. A.; Newton, J. M.; Davis, A. R.; Foster, M. L.

1981-01-01

A series of ice detection tests was performed on the shuttle external tank (ET) and on ET target samples using a 35/95 GHz instrumentation radiometer. Ice was formed using liquid nitrogen and water spray inside a test enclosure containing ET spray on foam insulation samples. During cryogenic fueling operations prior to the shuttle orbiter engine firing tests, ice was formed with freon and water over a one meter square section of the ET LOX tank. Data analysis was performed on the ice signatures, collected by the radiometer, using Georgia Tech computing facilities. Data analysis technique developed include: ice signature images of scanned ET target; pixel temperature contour plots; time correlation of target data with ice present versus no ice formation; and ice signature radiometric temperature statistical data, i.e., mean, variance, and standard deviation.

Using a Novel Transgenic Mouse Model to Study c-Myc Oncogenic Pathway in Castration Resistance and Chemoresistance of Prostate Cancer

DTIC Science & Technology

2015-10-01

epididymis tumors, and performed implantation of testosterone pellets to continue support prostate tumor growth. Our recent data suggest that these...will eliminate the concerns on epididymis tumors. We also proposed to perform subcutaneous implantation of testosterone pellets to continue...USAMRMC ACURO. We have initiated performing the early castration and testosterone pellet implantation on the young PB-Cre4/CAG- SMIL/p53loxP/loxP

Metal Matrix Composite LOX Turbopump Housing via Novel Tool-less Net-Shape Pressure Infiltration Casting Technology

NASA Technical Reports Server (NTRS)

Shah, Sandeep; Lee, Jonathan; Bhat, Biliyar; Wells, Doug; Gregg, Wayne; Marsh, Matthew; Genge, Gary; Forbes, John; Salvi, Alex; Cornie, James A.

2003-01-01

Metal matrix composites for propulsion components offer high performance and affordability, resulting in low weight and cost. The following sections in this viewgraph presentation describe the pressure infiltration casting of a metal matrix composite LOX turbopump housing: 1) Baseline Pump Design and Stress Analysis; 2) Tool-less Advanced Pressure Infiltration Casting Process; 3) Preform Splicing and Joining for Large Components such as Pump Housing; 4) Fullscale Pump Housing Redesign.

LOX tank installation

NASA Image and Video Library

2011-06-08

Construction of the A-3 Test Stand at Stennis Space Center continued June 8 with installation of a 35,000-gallon liquid oxygen tank atop the steel structure. The stand is being built to test next-generation rocket engines that will carry humans into deep space once more. The LOX tank and a liquid hydrogen tank to be installed atop the stand later will provide propellants for testing the engines. The A-3 Test Stand is scheduled for completion and activation in 2013.

Hypoxia triggers angiogenesis by increasing expression of LOX genes in 3-D culture of ASCs and ECs.

PubMed

Xie, Qiang; Xie, Jiamin; Tian, Taoran; Ma, Quanquan; Zhang, Qi; Zhu, Bofeng; Cai, Xiaoxiao

2017-03-01

This study aimed to investigate the expression changes of LOX (lysyl oxidase) family genes, VEGFA, and VEGFB under hypoxic conditions in a co-culture system of ASCs (adipose-derived stromal cells) and ECs (endothelial cells). ASCs and ECs were co-cultured under hypoxic and normal oxygen conditions in a 1:1 ratio, and the formation of vessel-like was detected at 7 days. The transwell co-culture system was used and cell lysates were collected at 7 days after co-culture in hypoxic and normal oxygen condition. Semi-quantitative PCR was performed to quantify the mRNA expression of VEGFA, VEGFB, and the LOX genes (LOX, LOXL-1, LOXL-2, LOXL-3, and LOXL-4). Expression changes were determined by densitomery. Enhanced angiogenesis was detected in the co-culture of ASCs and ECs under hypoxic conditions. Among the genes screened, VEGFA, VEGFB, LOXL-1, and LOXL-3 in ECs, both mono-cultured and co-cultured, were significantly enhanced after culturing under hypoxic conditions. In ASCs, VEGFB, LOXL-1, and LOXL-3 were upregulated. Contact co-culture between ASCs and ECs promotes angiogenesis under hypoxia. LOXL-1 and LOXL-3 expressions were increased in both ASCs and ECs and might play important roles in the enhanced angiogenesis promoted by hypoxia. Copyright © 2017 Elsevier Inc. All rights reserved.

Varietal Dependence of GLVs Accumulation and LOX-HPL Pathway Gene Expression in Four Vitis vinifera Wine Grapes

PubMed Central

Qian, Xu; Xu, Xiao-Qing; Yu, Ke-Ji; Zhu, Bao-Qing; Lan, Yi-Bin; Duan, Chang-Qing; Pan, Qiu-Hong

2016-01-01

Variety is one of the major factors influencing grape and wine aromatic characteristics. Green leaf volatiles (GLVs), derived from lipoxygenase-hydroperoxides lyase (LOX-HPL) pathway, are important components for the aromatic quality of grapes and wines. However, the varietal difference regarding GLVs accumulation and related gene expression are poorly studied. This work exhibited that the accumulation of various GLVs and the expression of LOX-HPL pathway genes in four Vitis vinifera wine grape cultivars: Syrah, Muscat Tchervine, Gewürztraminer and Chardonnay. The results showed a variety dependence of GLVs profile. Muscat Tchervine harvested grapes contained less C6 aldehydes and the most abundant esters, which corresponded to very low VvLOXA and VvHPL1 expression abundance as well as high VvAAT transcript in this variety. High expression level of both VvLOXA and VvHPL1 paralleled with higher level of C6 aldehydes together with higher alcohols in Syrah grape. Gewürztraminer and Chardonnay grapes had high aldehydes and alcohols as well as low esters, which were resulted from their higher expression level of VvLOXA or VvHPL1 and lower VvAAT. From these above corresponding relations, it is concluded that VvLOXA, VvHPL1 and VvAAT in the LOX-HPL pathway are targets for altering GLVs composition in the grape varieties. PMID:27886056

Isolation and characterization of 9-lipoxygenase and epoxide hydrolase 2 genes: Insight into lactone biosynthesis in mango fruit (Mangifera indica L.).

PubMed

Deshpande, Ashish B; Chidley, Hemangi G; Oak, Pranjali S; Pujari, Keshav H; Giri, Ashok P; Gupta, Vidya S

2017-06-01

Uniqueness and diversity of mango flavour across various cultivars are well known. Among various flavour metabolites lactones form an important class of aroma volatiles in certain mango varieties due to their ripening specific appearance and lower odour detection threshold. In spite of their biological and biochemical importance, lactone biosynthetic pathway in plants remains elusive. Present study encompasses quantitative real-time analysis of 9-lipoxygenase (Mi9LOX), epoxide hydrolase 2 (MiEH2), peroxygenase, hydroperoxide lyase and acyl-CoA-oxidase genes during various developmental and ripening stages in fruit of Alphonso, Pairi and Kent cultivars with high, low and no lactone content and explains their variable lactone content. Study also covers isolation, recombinant protein characterization and transient over-expression of Mi9LOX and MiEH2 genes in mango fruits. Recombinant Mi9LOX utilized linoleic and linolenic acids, while MiEH2 utilized aromatic and fatty acid epoxides as their respective substrates depicting their role in fatty acid metabolism. Significant increase in concentration of δ-valerolactone and δ-decalactone upon Mi9LOX over-expression and that of δ-valerolactone, γ-hexalactone and δ-hexalactone upon MiEH2 over-expression further suggested probable involvement of these genes in lactone biosynthesis in mango. Copyright © 2017 Elsevier Ltd. All rights reserved.

Inhibition of in vitro leukotriene B4 biosynthesis in human neutrophil granulocytes and docking studies of natural quinones.

PubMed

Landa, Premysl; Kutil, Zsofia; Temml, Veronika; Malik, Jan; Kokoska, Ladislav; Widowitz, Ute; Pribylova, Marie; Dvorakova, Marcela; Marsik, Petr; Schuster, Daniela; Bauer, Rudolf; Vanek, Tomas

2013-01-01

Quinones are compounds frequently contained in medicinal plants used for the treatment of inflammatory diseases. Therefore, the impact of plant-derived quinones on the arachidonic acid metabolic pathway is worthy of investigation. In this study, twenty-three quinone compounds of plant origin were tested in vitro for their potential to inhibit leukotriene B4 (LTB4) biosynthesis in activated human neutrophil granulocytes with 5-lipoxygenase (5-LOX) activity. The benzoquinones primin (3) and thymohydroquinone (4) (IC50 = 4.0 and 4.1 microM, respectively) showed activity comparable with the reference inhibitor zileuton (1C50 = 4.1 microM). Moderate activity was observed for the benzoquinone thymoquinone (2) (1C50 = 18.2 microM) and the naphthoquinone shikonin (1) (IC50 = 24.3 microM). The anthraquinone emodin and the naphthoquinone plumbagin (5) displayed only weak activities (IC50 > 50 microM). The binding modes of the active compounds were further evaluated in silico by molecular docking to the human 5-LOX crystal structure. This process supports the biological data and suggested that, although the redox potential is responsible for the quinone's activity on multiple targets, in the case of 5-LOX the molecular structure plays a vital role in the inhibition. The obtained results suggest primin as a promising compound for the development of dual COX-2/5-LOX inhibitors.

Cre/lox system to develop selectable marker free transgenic tobacco plants conferring resistance against sap sucking homopteran insect.

PubMed

Chakraborti, Dipankar; Sarkar, Anindya; Mondal, Hossain A; Schuermann, David; Hohn, Barbara; Sarmah, Bidyut K; Das, Sampa

2008-10-01

A binary expression vector was constructed containing the insecticidal gene Allium sativum leaf agglutinin (ASAL), and a selectable nptII marker gene cassette, flanked by lox sites. Similarly, another binary vector was developed with the chimeric cre gene construct. Transformed tobacco plants were generated with these two independent vectors. Each of the T(0) lox plants was crossed with T(0) Cre plants. PCR analyses followed by the sequencing of the target T-DNA part of the hybrid T(1) plants demonstrated the excision of the nptII gene in highly precised manner in certain percentage of the T(1) hybrid lines. The frequency of such marker gene excision was calculated to be 19.2% in the hybrids. Marker free plants were able to express ASAL efficiently and reduce the survivability of Myzus persiceae, the deadly pest of tobacco significantly, compared to the control tobacco plants. Results of PCR and Southern blot analyses of some of the T(2) plants detected the absence of cre as well as nptII genes. Thus, the crossing strategy involving Cre/lox system for the excision of marker genes appears to be very effective and easy to execute. Documentation of such marker excision phenomenon in the transgenic plants expressing the important insecticidal protein for the first time has a great significance from agricultural and biotechnological points of view.

Flow Distribution Around the SSME Main Injector Assembly Using Porosity Formulation

NASA Technical Reports Server (NTRS)

Cheng, Gary C.; Chen, Yen-Sen; Wang, Ten-See

1995-01-01

Hot gas turbulent flow distribution around the main injector assembly of the Space Shuttle Main Engine (SSME) and Liquid Oxidizer (LOX) flow distribution through the LOX posts have a great effect on the combustion phenomena inside the main combustion chamber. In order to design a CFD model to be an effective engineering analysis tool with good computational turn- around time (especially for 3-D flow problems) and still maintain good accuracy in describing the flow features, the concept of porosity was employed to describe the effects of blockage and drag force due to the presence of the LOX posts in the turbulent flow field around the main injector assembly of the SSME. 2-D numerical studies were conducted to identify the drag coefficients of the flows both through tube banks and around the shielded posts over a wide range of Reynolds numbers. Empirical, analytical expressions of the drag coefficient as a function of local flow Reynolds number were then deduced. The porosity model was applied to the turbulent flow around the main injector assembly of the SSME, and analyses were performed. The 3-D CFD analysis was divided into three parts, LOX dome, hot gas injector assembly, and hydrogen cavity. The numerical results indicate that the mixture ratio at the downstream of injector face was close to stoichiometric around baffle elements.

Modulation of Cox-1, 5-, 12- and 15-Lox by popular herbal remedies used in southern Italy against psoriasis and other skin diseases.

PubMed

Bader, Ammar; Martini, Francesca; Schinella, Guillermo R; Rios, Jose L; Prieto, Jose M

2015-01-01

Acanthus mollis (Acanthaceae), Achillea ligustica, Artemisia arborescens and Inula viscosa (Asteraceae) are used in Southern Italy against psoriasis and other skin diseases that occur with an imbalanced production of eicosanoids. We here assessed their in vitro effects upon 5-, 12-, 15-LOX and COX-1 enzymes as well as NFκB activation in intact cells as their possible therapeutic targets. All methanol crude extracts inhibited both 5-LOX and COX-1 activities under 200 µg/mL, without significant effects on the 12-LOX pathway or any relevant in vitro free radical scavenging activity. NFκB activation was prevented by all extracts but A. mollis. Interestingly, A. ligustica, A. arborescens and A. mollis increased the biosynthesis of 15(S)-HETE, an anti-inflammatory eicosanoid. A. ligustica (IC50 =49.5 µg/mL) was superior to Silybum marianum (IC50 =147.8 µg/mL), which we used as antipsoriatic herbal medicine of reference. Its n-hexane, dichloromethane and ethyl acetate fractions had also inhibitory effects on the LTB4 biosynthesis (IC50 s=9.6, 20.3 and 68 µg/mL, respectively) evidencing that the apolar extracts of A. ligustica are promising active herbal ingredients for future phytotherapeutical products targeting psoriasis. © 2014 The Authors. Phytotherapy Research published by John Wiley & Sons Ltd.

Flow Distribution Around the SSME Main Injector Assembly Using Porosity Formulation

NASA Technical Reports Server (NTRS)

Cheng, Gary C.; Chen, Yen-Sen; Wang, Ten-See

1995-01-01

Hot gas turbulent flow distribution around the main injector assembly of the Space Shuttle Main Engine (SSME) and Liquid Oxidizer (LOX) flow distribution through the LOX posts have a great effect on the combustion phenomena inside the main combustion chamber. In order to design a CFD model to be an effective engineering analysis tool with good computational turn-around time (especially for 3-D flow problems) and still maintain good accuracy in describing the flow features, the concept of porosity was employed to describe the effects of blockage and drag force due to the presence of the LOX posts in the turbulent flow field around the main injector assembly of the SSME. 2-D numerical studies were conducted to identify the drag coefficients of the flows both through tube banks and around the shielded posts over a wide range of Reynolds numbers. Empirical, analytical expressions of the drag coefficient as a function of local flow Reynolds number were then deduced. The porosity model was applied to the turbulent flow around the main injector assembly of the SSME, and analyses were performed. The 3-D CFD analysis was divided into three parts, LOX dome, hot gas injector assembly, and hydrogen cavity. The numerical results indicate that the mixture ratio at the downstream of injector face was close to stoichiometric around baffle elements.

Vinpocetine attenuates lipid accumulation and atherosclerosis formation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cai, Yujun; Li, Jian-Dong; Yan, Chen, E-mail: Chen_Yan@urmc.rochester.edu

Highlights: •Vinpocetine attenuates hyperlipidemia-induced atherosclerosis in a mouse model. •Vinpocetine antagonizes ox-LDL uptake and accumulation in macrophages. •Vinpocetine blocks the induction of ox-LDL receptor LOX-1 in vitro and in vivo. -- Abstract: Atherosclerosis, the major cause of myocardial infarction and stroke, is a chronic arterial disease characterized by lipid deposition and inflammation in the vessel wall. Cholesterol, in low-density lipoprotein (LDL), plays a critical role in the pathogenesis of atherosclerosis. Vinpocetine, a derivative of the alkaloid vincamine, has long been used as a cerebral blood flow enhancer for treating cognitive impairment. Recent study indicated that vinpocetine is a potent anti-inflammatorymore » agent. However, its role in the pathogenesis of atherosclerosis remains unexplored. In the present study, we show that vinpocetine significantly reduced atherosclerotic lesion formation in ApoE knockout mice fed with a high-fat diet. In cultured murine macrophage RAW264.7 cells, vinpocetine markedly attenuated oxidized LDL (ox-LDL) uptake and foam cell formation. Moreover, vinpocetine greatly blocked the induction of ox-LDL receptor 1 (LOX-1) in cultured macrophages as well as in the LOX-1 level in atherosclerotic lesions. Taken together, our data reveal a novel role of vinpocetine in reduction of pathogenesis of atherosclerosis, at least partially through suppressing LOX-1 signaling pathway. Given the excellent safety profile of vinpocetine, this study suggests vinpocetine may be a therapeutic candidate for treating atherosclerosis.« less

A facile one-step route to synthesize cage-like silica hollow spheres loaded with superparamagnetic iron oxide nanoparticles in their shells.

PubMed

Li, Ling; Choo, Eugene Shi Guang; Tang, Xiaosheng; Ding, Jun; Xue, Junmin

2009-02-28

Cage-like silica hollow spheres loaded with superparamagnetic iron oxide nanoparticles incorporated in their macroporous shells are synthesized in a facile manner through a one-step oil-in-diethylene glycol (DEG) microemulsion route.

Inhibition of lipoxygenases and cyclooxygenase-2 enzymes by extracts isolated from Bacopa monniera (L.) Wettst.

PubMed

Viji, V; Helen, A

2008-07-23

Bacopa monniera Linn is described in the Ayurvedic Materia Medica, as a therapeutically useful herb for the treatment of inflammation. In the current study, we investigated the anti-inflammatory activity of methanolic extract of Bacopa monniera (BME). For some experiments EtOAc and bacoside fractions were prepared from BME. The effect of these extracts in modulating key mediators of inflammation was evaluated. Carrageenan-induced rat paw edema, rat mononuclear cells and human whole blood assay were employed as in vivo and in vitro models. In carrageenan-induced rat paw edema, BME brought about 82% edema inhibition at a dose of 100mg/kg i.p. when compared to indomethacin (INDO) (3mg/kg) that showed 70% edema inhibition. BME also significantly inhibited 5-lipoxygenase (5-LOX), 15-LOX and cyclooxygenase-2 (COX-2) activities in rat monocytes in vivo. Among the fractions tested in vitro, EtOAc fraction possessed significant 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity with IC(50) value of 30 microg/ml compared to butylated hydroxyl toluene (IC(50) = 13 microg/ml). This fraction also exerted significant hydroxyl radical scavenging activity with IC(50) value of 25 microg/ml in comparison with quercetin (IC(50) = 5 microg/ml). Inhibitory effects of EtOAc and bacoside fractions on LOX and COX activities in Ca-A23187 stimulated rat mononuclear cells were also assessed. 5-LOX IC(50) values were 25 microg/ml for EtOAc, 68 microg/ml for bacosides and 2 microg/ml for nordihydroguaiaretic acid (NDGA) where as COX-2 IC(50) values were 1.32 microg/ml for EtOAc, 1.19 microg/ml for bacoside fraction and 0.23 microg/ml for indomethacin. EtOAc and bacoside fractions also brought about significant decrease in TNF-alpha release ex vivo. Bacopa monniera possesses anti-inflammatory activity through inhibition of COX and LOX and downregulation of TNF-alpha.

Inhibition of 12/15-Lipoxygenase Protects Against β-Cell Oxidative Stress and Glycemic Deterioration in Mouse Models of Type 1 Diabetes.

PubMed

Hernandez-Perez, Marimar; Chopra, Gaurav; Fine, Jonathan; Conteh, Abass M; Anderson, Ryan M; Linnemann, Amelia K; Benjamin, Chanelle; Nelson, Jennifer B; Benninger, Kara S; Nadler, Jerry L; Maloney, David J; Tersey, Sarah A; Mirmira, Raghavendra G

2017-11-01

Islet β-cell dysfunction and aggressive macrophage activity are early features in the pathogenesis of type 1 diabetes (T1D). 12/15-Lipoxygenase (12/15-LOX) is induced in β-cells and macrophages during T1D and produces proinflammatory lipids and lipid peroxides that exacerbate β-cell dysfunction and macrophage activity. Inhibition of 12/15-LOX provides a potential therapeutic approach to prevent glycemic deterioration in T1D. Two inhibitors recently identified by our groups through screening efforts, ML127 and ML351, have been shown to selectively target 12/15-LOX with high potency. Only ML351 exhibited no apparent toxicity across a range of concentrations in mouse islets, and molecular modeling has suggested reduced promiscuity of ML351 compared with ML127. In mouse islets, incubation with ML351 improved glucose-stimulated insulin secretion in the presence of proinflammatory cytokines and triggered gene expression pathways responsive to oxidative stress and cell death. Consistent with a role for 12/15-LOX in promoting oxidative stress, its chemical inhibition reduced production of reactive oxygen species in both mouse and human islets in vitro. In a streptozotocin-induced model of T1D in mice, ML351 prevented the development of diabetes, with coincident enhancement of nuclear Nrf2 in islet cells, reduced β-cell oxidative stress, and preservation of β-cell mass. In the nonobese diabetic mouse model of T1D, administration of ML351 during the prediabetic phase prevented dysglycemia, reduced β-cell oxidative stress, and increased the proportion of anti-inflammatory macrophages in insulitis. The data provide the first evidence to date that small molecules that target 12/15-LOX can prevent progression of β-cell dysfunction and glycemic deterioration in models of T1D. © 2017 by the American Diabetes Association.

Changes of oxidase and hydrolase activities in pecan leaves elicited by black pecan aphid (Hemiptera: Aphididae) feeding.

PubMed

Chen, Yigen; Ni, Xinzhi; Cottrell, Ted E; Wood, Bruce W; Buntin, G David

2009-06-01

The black pecan aphid, Melanocallis caryaefoliae (Davis) (Hemiptera: Aphididae), is a foliar feeder of pecan, Carya illinoinensis (Wangenh.) K. Koch (Juglandaceae). The pest causes chlorosis of leaflet lamina, physiological damage to foliage and trees, and commonly limits the profitability of commercial pecan orchard enterprises. However, key aspects of this host-pest interaction are poorly understood. We report here the effects of M. caryaefoliae feeding on the foliar activity of oxidative (i.e., catalase, lipoxygenase [LOX]-1 and 3, and peroxidase) and hydrolytic (i.e., esterase) enzymes in relation to the degree of aphid resistance among pecan varieties. The 2-yr study showed that M. caryaefoliae-infested foliage exhibited elevated peroxidase activity only in susceptible ('Desirable', 'Sumner', and 'Schley'), but not in resistant ('Cape Fear', 'Gloria Grande', and 'Money Maker') genotypes. Susceptible genotypes also exhibited more severe leaf chlorosis in response to M. caryaefoliae feeding than the resistant genotypes; however, the aphid feeding did not influence catalase or esterase activity in all varieties, except the increase of esterase activity in Desirable and Gloria Grande. Melanocallis caryaefoliae feeding also influences activity of two lipoxygenase isozymes, with LOX3 being more frequently induced than LOX1. Foliar LOX3 activity was more frequently induced by M. caryaefoliae feeding in the moderately resistant 'Oconee' and highly resistant Money Maker and Cape Fear than in the susceptible genotypes. Therefore, the elevation of peroxidase is likely to be associated with aphid susceptibility and contributed to the severe leaf chlorosis, whereas the increase of LOX3 activity might be associated with aphid resistance in pecan. These findings contribute to our understanding of the etiology of M. caryaefoliae-elicited leaf chlorosis on pecan foliage. Such information may also be used to develop enzyme markers for identifying black pecan aphid resistance and/or susceptibility in pecan germplasm.

The eicosanoid response to high dose UVR exposure of individuals prone and resistant to sunburn.

PubMed

Nicolaou, Anna; Masoodi, Mojgan; Gledhill, Karl; Haylett, Ann Katarina; Thody, Anthony John; Tobin, Desmond John; Rhodes, Lesley Elizabeth

2012-02-01

High personal UVR doses can be gained during leisure activities, causing intense self-resolving inflammation (sunburn) of unprotected skin. UVR activates release of membrane fatty acids and upregulates their metabolism by cyclooxygenases (COX) and lipoxygenases (LOX) to different eicosanoids. While COX-derived prostaglandin (PG)E(2) is a potent mediator of sunburn vasodilatation, LOX-derived 15-hydroxyeicosatetraenoic acid (HETE) and its lipoxin metabolites may contribute to sunburn limitation. We explored the relationships between expression of these lipid mediators and the clinical and histological outcomes, comparing responses of individuals prone and more resistant to sunburn. An acute UVR exposure of 12 SED (standard erythema dose) was applied to buttock skin of 32 white Caucasians (n = 16 phototype I/II, n = 16 phototype III/IV), and over the subsequent 72 h assessments were made of skin erythema, immunohistochemical expression of leukocyte markers, COX-2, 12-LOX, 15-LOX and nitric oxide synthase (NOS), and eicosanoid levels by LC/ESI-MS/MS. Evidence of a significant inflammatory response was seen earlier in phototype I/II with regard to expression of erythema (4 h, p < 0.001), neutrophil infiltration (24 h, p = 0.01), epidermal COX-2 (24 h, p < 0.05) and 12-LOX (24 h, p < 0.01), and dermal eNOS (24 h, p < 0.05) proteins, although CD3+ lymphocyte infiltration showed an earlier increase in phototype III/IV (24 h, p < 0.05). Although erythema was equivalent at 72 h in both groups, phototype I/II showed higher PGE(2) accompanied by elevated 15-HETE, and a strong positive correlation was seen between these mediators (n = 18, r = 0.805, p = 0.0001). Hence anti-inflammatory eicosanoid 15-HETE may temper the pro-inflammatory milieu in sunburn, having greater influence in those prone to sunburn than those more resistant, given the same high UVR exposure conditions. This journal is © The Royal Society of Chemistry and Owner Societies 2012

Large Impact of Low Concentration Oxidized LDL on Angiogenic Potential of Human Endothelial Cells: A Microarray Study

PubMed Central

Khaidakov, Magomed; Mitra, Sona; Wang, Xianwei; Ding, Zufeng; Bora, Nalini; Lyzogubov, Valery; Romeo, Francesco; Schichman, Steven A.; Mehta, Jawahar L.

2012-01-01

Oxidized LDL (ox-LDL) is a key factor in atherogenesis. It is taken up by endothelial cells primarily by ox-LDL receptor-1 (LOX-1). To elucidate transcriptional responses, we performed microarray analysis on human coronary artery endothelial cells (HCAECs) exposed to small physiologic concentration of ox-LDL- 5 µg/ml for 2 and 12 hours. At 12 hours, cultures treated with ox-LDL exhibited broad shifts in transcriptional activity involving almost 1500 genes (>1.5 fold difference, p<0.05). Resulting transcriptome was enriched for genes associated with cell adhesion (p<0.002), angiogenesis (p<0.0002) and migration (p<0.006). Quantitative PCR analysis revealed that LOX-1 expression in HCAECs is at least an order of magnitude greater than the expression of other major ox-LDL specific receptors CD36 and MSR1. In keeping with the data on LOX-1 expression, pre-treatment of HCAECs with LOX-1 neutralizing antibody resulted in across-the-board inhibition of cellular response to ox-LDL. Ox-LDL upregulated a number of pro-angiogenic genes including multiple receptors, ligands and transcription factors and altered the expression of a number of genes implicated in both stimulation and inhibition of apoptosis. From a functional standpoint, physiologic concentrations of ox-LDL stimulated tube formation and inhibited susceptibility to apoptosis in HCAECs. In addition, ox-LDL exposure resulted in upregulation of miR-1974, miR-1978 and miR-21 accompanied with significant over-presentation of their target genes in the downregulated portion of ox-LDL transcriptome. Our observations indicate that ox-LDL at physiologic concentrations induces broad transcriptional responses which are mediated by LOX-1, and are, in part, shaped by ox-LDL-dependent miRNAs. We also suggest that angiogenic effects of ox-LDL are partially based on upregulation of several receptors that render cells hypersensitive to angiogenic stimuli. PMID:23115646

Low Cost Propulsion Technology Testing at the Stennis Space Center: Propulsion Test Article and the Horizontal Test Facility

NASA Technical Reports Server (NTRS)

Fisher, Mark F.; King, Richard F.; Chenevert, Donald J.

1998-01-01

The need for low cost access to space has initiated the development of low cost liquid rocket engine and propulsion system hardware at the Marshall Space Flight Center. This hardware will be tested at the Stennis Space Center's B-2 test stand. This stand has been reactivated for the testing of the Marshall designed Fastrac engine and the Propulsion Test Article. The RP-1 and LOX engine is a turbopump fed gas generator rocket with an ablative nozzle which has a thrust of 60,000 lbf. The Propulsion Test Article (PTA) is a test bed for low cost propulsion system hardware including a composite RP-I tank, flight feedlines and pressurization system, stacked in a booster configuration. The PTA is located near the center line of the B-2 test stand, firing vertically into the water cooled flame deflector. A new second position on the B-2 test stand has been designed and built for the horizontal testing of the Fastrac engine in direct support of the X-34 launch vehicle. The design and integration of these test facilities as well as the coordination which was required between the two Centers is described and lessons learned are provided. The construction of the horizontal test position is discussed in detail. The activation of these facilities is examined and the major test milestones are described.

Pressure fed thrust chamber technology program

NASA Technical Reports Server (NTRS)

Dunn, Glenn M.

1992-01-01

This is the final report for the Pressure Fed Technology Program. It details the design, fabrication and testing of subscale hardware which successfully characterized LOX/RP combustion for a low cost pressure fed design. The innovative modular injector design is described in detail as well as hot-fire test results which showed excellent performance. The program summary identifies critical LOX/RP design issues that have been resolved by this testing, and details the low risk development requirements for a low cost engine for future Expendable Launch Vehicles (ELVi).

Saturn V First Stage S-1C LOX Fuel Tanks

NASA Technical Reports Server (NTRS)

1960-01-01

This photograph shows the Saturn V assembled LOX (Liquid Oxygen) and fuel tanks ready for transport from the Manufacturing Engineering Laboratory at Marshall Space Flight Center in Huntsville, Alabama. The tanks were then shipped to the launch site at Kennedy Space Center for a flight. The towering 363-foot Saturn V was a multi-stage, multi-engine launch vehicle standing taller than the Statue of Liberty. Altogether, the Saturn V engines produced as much power as 85 Hoover Dams.

A New High-Speed, High-Cycle, Gear-Tooth Bending Fatigue Test Capability

NASA Technical Reports Server (NTRS)

Stringer, David B.; Dykas, Brian D.; LaBerge, Kelsen E.; Zakrajsek, Andrew J.; Handschuh, Robert F.

2011-01-01

A new high-speed test capability for determining the high cycle bending-fatigue characteristics of gear teeth has been developed. Experiments were performed in the test facility using a standard spur gear test specimens designed for use in NASA Glenn s drive system test facilities. These tests varied in load condition and cycle-rate. The cycle-rate varied from 50 to 1000 Hz. The loads varied from high-stress, low-cycle loads to near infinite life conditions. Over 100 tests were conducted using AISI 9310 steel spur gear specimen. These results were then compared to previous data in the literature for correlation. Additionally, a cycle-rate sensitivity analysis was conducted by grouping the results according to cycle-rate and comparing the data sets. Methods used to study and verify load-path and facility dynamics are also discussed.

Flow Quality for Turbine Engine Loads Simulator (TELS) Facility

DTIC Science & Technology

1980-06-01

2.2 GAS INGESTION A mathematical simulation of the turbojet engine and jet deflector was formulated to estimate the severity of the recirculating...3. Swain. R. L. and Mitchell, J. G. "’Smlulatlon of Turbine Engine Operational Loads." Journal of Aircraft Vol. 15, No. 6, June 1978• 4. Ryan, J...3 AEDC-TR-79-83 ~...~ i ,i g - Flow Quality for Turbine Engine Loads Simulator (TELS) Facility R..I. Schulz ARO, Inc. June 1980

CFD Validation with Experiment and Verification with Physics of a Propellant Damping Device

NASA Technical Reports Server (NTRS)

Yang, H. Q.; Peugeot, John

2011-01-01

This paper will document our effort in validating a coupled fluid-structure interaction CFD tool in predicting a damping device performance in the laboratory condition. Consistently good comparisons of "blind" CFD predictions against experimental data under various operation conditions, design parameters, and cryogenic environment will be presented. The power of the coupled CFD-structures interaction code in explaining some unexpected phenomena of the device observed during the technology development will be illustrated. The evolution of the damper device design inside the LOX tank will be used to demonstrate the contribution of the tool in understanding, optimization and implementation of LOX damper in Ares I vehicle. It is due to the present validation effort, the LOX damper technology has matured to TRL 5. The present effort has also contributed to the transition of the technology from an early conceptual observation to the baseline design of thrust oscillation mitigation for the Ares I within a 10 month period.

A lactate electrochemical biosensor with a titanate nanotube as direct electron transfer promoter

NASA Astrophysics Data System (ADS)

Yang, Mingli; Wang, Jin; Li, Huaqing; Zheng, Jian-Guo; Wu, Nianqiang Nick

2008-02-01

Hydrogen titanate (H2Ti3O7) nanotubes (TNTs) have been synthesized by a one-step hydrothermal processing. Lactate oxidase (LOx) enzyme has been immobilized on the three-dimensional porous TNT network to make an electrochemical biosensor for lactate detection. Cyclic voltammetry and amperometry tests reveal that the LOx enzyme, which is supported on TNTs, maintains their substrate-specific catalytic activity. The nanotubes offer the pathway for direct electron transfer between the electrode surface and the active redox centers of LOx, which enables the biosensor to operate at a low working potential and to avoid the influence of the presence of O2 on the amperometric current response. The biosensor exhibits a sensitivity of 0.24 µA cm-2 mM-1, a 90% response time of 5 s, and a linear response in the range from 0.5 to 14 mM and the redox center of enzyme obviates the need of redox mediators for electrochemical enzymatic sensors, which is attractive for the development of reagentless biosensors.

Structure-based design, synthesis, molecular docking study and biological evaluation of 1,2,4-triazine derivatives acting as COX/15-LOX inhibitors with anti-oxidant activities.

PubMed

Khoshneviszadeh, Mehdi; Shahraki, Omolbanin; Khoshneviszadeh, Mahsima; Foroumadi, Alireza; Firuzi, Omidreza; Edraki, Najmeh; Nadri, Hamid; Moradi, Alireza; Shafiee, Abbas; Miri, Ramin

2016-12-01

A set of 1,2,4-triazine derivatives were designed as cyclooxygenase-2 (COX-2) inhibitors. These compounds were synthesized and screened for inhibition of cyclooxygenases (COX-1 and COX-2) based on a cellular assay using human whole blood (HWB) and lipoxygenase (LOX-15) that are key enzymes in inflammation. The results showed that 3-(2-(benzo[d][1,3]dioxol-5-ylmethylene)hydrazinyl)-5,6-bis(4-methoxyphenyl)-1,2,4-triazine (G11) was identified as the most potent COX-2 inhibitor (78%) relative to COX-1 (50%). Ferric reducing anti-oxidant power (FRAP) assay revealed that compound G10 possesses the highest anti-oxidant activity. The compound G3 with IC50 value of 124 μM was the most potent compound in LOX inhibitory assay. Molecular docking was performed and a good agreement was observed between computational and experimental results.

LOX/LH2 vane pump for auxiliary propulsion systems

NASA Technical Reports Server (NTRS)

Hemminger, J. A.; Ulbricht, T. E.

1985-01-01

Positive displacement pumps offer potential efficiency advantages over centrifugal pumps for future low thrust space missions. Low flow rate applications, such as space station auxiliary propulsion or dedicated low thrust orbiter transfer vehicles, are typical of missions where low flow and high head rise challenge centrifugal pumps. The positive displacement vane pump for pumping of LOX and LH2 is investigated. This effort has included: (1) a testing program in which pump performance was investigated for differing pump clearances and for differing pump materials while pumping LN2, LOX, and LH2; and (2) an analysis effort, in which a comprehensive pump performance analysis computer code was developed and exercised. An overview of the theoretical framework of the performance analysis computer code is presented, along with a summary of analysis results. Experimental results are presented for pump operating in liquid nitrogen. Included are data on the effects on pump performance of pump clearance, speed, and pressure rise. Pump suction performance is also presented.

Time-Resolved Investigations of Heterobimetallic Cofactor Assembly in R2lox Reveal Distinct Mn/Fe Intermediates.

PubMed

Miller, Effie K; Trivelas, Nicholas E; Maugeri, Pearson T; Blaesi, Elizabeth J; Shafaat, Hannah S

2017-07-05

The assembly mechanism of the Mn/Fe ligand-binding oxidases (R2lox), a family of proteins that are homologous to the nonheme diiron carboxylate enzymes, has been investigated using time-resolved techniques. Multiple heterobimetallic intermediates that exhibit unique spectral features, including visible absorption bands and exceptionally broad electron paramagnetic resonance signatures, are observed through optical and magnetic resonance spectroscopies. On the basis of comparison to known diiron species and model compounds, the spectra have been attributed to (μ-peroxo)-Mn III /Fe III and high-valent Mn/Fe species. Global spectral analysis coupled with isotopic substitution and kinetic modeling reveals elementary rate constants for the assembly of Mn/Fe R2lox under aerobic conditions. A complete reaction mechanism for cofactor maturation that is consistent with experimental data has been developed. These results suggest that the Mn/Fe cofactor can perform direct C-H bond abstraction, demonstrating the potential for potent chemical reactivity that remains unexplored.

Polyamines induce adaptive responses in water deficit stressed cucumber roots.

PubMed

Kubiś, Jan; Floryszak-Wieczorek, Jolanta; Arasimowicz-Jelonek, Magdalena

2014-01-01

The aim of this study was to investigate the effect of exogenous polyamines (PAs) on the membrane status and proline level in roots of water stressed cucumber (Cucumis sativus cv. Dar) seedlings. It was found that water shortage resulted in an increase of membrane injury, lipoxygenase (LOX) activity, lipid peroxidation and proline concentration in cucumber roots during progressive dehydration. PA pretreatment resulted in a distinct reduction of the injury index, and this effect was reflected by a lower stress-evoked LOX activity increase and lipid peroxide levels at the end of the stress period. In contrast, PA-supplied stressed roots displayed a higher proline accumulation. The presented results suggest that exogenous PAs are able to alleviate water deficit-induced membrane permeability and diminish LOX activity. Observed changes were accompanied by an accumulation of proline, suggesting that the accumulation of this osmolyte might be another possible mode of action for PAs to attain higher membrane stability, and in this way mitigate water deficit effects in roots of cucumber seedlings.

Multifunctional Cinnamic Acid Derivatives.

PubMed

Peperidou, Aikaterini; Pontiki, Eleni; Hadjipavlou-Litina, Dimitra; Voulgari, Efstathia; Avgoustakis, Konstantinos

2017-07-25

Our research to discover potential new multitarget agents led to the synthesis of 10 novel derivatives of cinnamic acids and propranolol, atenolol, 1-adamantanol, naphth-1-ol, and (benzylamino) ethan-1-ol. The synthesized molecules were evaluated as trypsin, lipoxygenase and lipid peroxidation inhibitors and for their cytotoxicity. Compound 2b derived from phenoxyphenyl cinnamic acid and propranolol showed the highest lipoxygenase (LOX) inhibition (IC 50 = 6 μΜ) and antiproteolytic activity (IC 50 = 0.425 μΜ). The conjugate 1a of simple cinnamic acid with propranolol showed the higher antiproteolytic activity (IC 50 = 0.315 μΜ) and good LOX inhibitory activity (IC 50 = 66 μΜ). Compounds 3a and 3b , derived from methoxylated caffeic acid present a promising combination of in vitro inhibitory and antioxidative activities. The S isomer of 2b also presented an interesting multitarget biological profile in vitro . Molecular docking studies point to the fact that the theoretical results for LOX-inhibitor binding are identical to those from preliminary in vitro study.

Tissue-specifically regulated site-specific excision of selectable marker genes in bivalent insecticidal, genetically-modified rice.

PubMed

Hu, Zhan; Ding, Xuezhi; Hu, Shengbiao; Sun, Yunjun; Xia, Liqiu

2013-12-01

Marker-free, genetically-modified rice was created by the tissue-specifically regulated Cre/loxP system, in which the Cre recombinase gene and hygromycin phosphotransferase gene (hpt) were flanked by two directly oriented loxP sites. Cre expression was activated by the tissue-specific promoter OsMADS45 in flower or napin in seed, resulting in simultaneous excision of the recombinase and marker genes. Segregation of T1 progeny was performed to select recombined plants. The excision was confirmed by PCR, Southern blot and sequence analyses indicating that efficiency varied from 10 to 53 % for OsMADS45 and from 12 to 36 % for napin. The expression of cry1Ac and vip3A was detected by RT-PCR analysis in marker-free transgenic rice. These results suggested that our tissue-specifically regulated Cre/loxP system could auto-excise marker genes from transgenic rice and alleviate public concerns about the security of GM crops.

c-Kit-Mediated Functional Positioning of Stem Cells to Their Niches Is Essential for Maintenance and Regeneration of Adult Hematopoiesis

PubMed Central

Kimura, Yuki; Ding, Bisen; Imai, Norikazu; Nolan, Daniel J.; Butler, Jason M.; Rafii, Shahin

2011-01-01

The mechanism by which hematopoietic stem and progenitor cells (HSPCs) through interaction with their niches maintain and reconstitute adult hematopoietic cells is unknown. To functionally and genetically track localization of HSPCs with their niches, we employed novel mutant loxPs, lox66 and lox71 and Cre-recombinase technology to conditionally delete c-Kit in adult mice, while simultaneously enabling GFP expression in the c-Kit-deficient cells. Conditional deletion of c-Kit resulted in hematopoietic failure and splenic atrophy both at steady state and after marrow ablation leading to the demise of the treated adult mice. Within the marrow, the c-Kit-expressing GFP+ cells were positioned to Kit ligand (KL)-expressing niche cells. This c-Kit-mediated cellular adhesion was essential for long-term maintenance and expansion of HSPCs. These results lay the foundation for delivering KL within specific niches to maintain and restore hematopoiesis. PMID:22046410

Investigation of characteristics of feed system instabilities

NASA Technical Reports Server (NTRS)

Vaage, R. D.; Fidler, L. E.; Zehnle, R. A.

1972-01-01

The relationship between the structural and feed system natural frequencies in structure-propulsion system coupled longitudinal oscillations (pogo) is investigated. The feed system frequencies are usually very dependent upon the compressibility (compliance) of cavitation bubbles that exist to some extent in all operating turbopumps. This document includes: a complete review of cavitation mechanisms; development of a turbopump cavitation compliance model; an accumulation and analysis of all available cavitation compliance test data; and a correlation of empirical-analytical results. The analytical model is based on the analysis of flow relative to a set of cascaded blades, having any described shape, and assumes phase changes occur under conditions of isentropic equilibrium. Analytical cavitation compliance predictions for the J-2 LOX, F-1 LOX, H-1 LOX and LR87 oxidizer turbopump inducers do not compare favorably with test data. The model predicts much less cavitation than is derived from the test data. This implies that mechanisms other than blade cavitation contribute significantly to the total amount of turbopump cavitation.

Molecular simulations enlighten the binding mode of quercetin to lipoxygenase-3.

PubMed

Fiorucci, Sébastien; Golebiowski, Jérôme; Cabrol-Bass, Daniel; Antonczak, Serge

2008-11-01

Inhibition of lipoxygenases (LOXs) by flavonoid compounds is now well documented, but the description of the associated mechanism remains controversial due to a lack of information at the molecular level. For instance, X-ray determination of quercetin/LOX-3 system has led to a structure where the enzyme was cocrystallized with a degradation product of the substrate, which rendered the interpretation of the reported interactions between this flavonoid compound and the enzyme difficult. Molecular modeling simulations can in principle allow obtaining precious insights that could fill this lack of structural information. Thus, in this study, we have investigated various binding modes of quercetin to LOX-3 enzyme in order to understand the first step of the inhibition process, that is the association of the two entities. Molecular dynamics simulations and free energy calculations suggest that quercetin binds the metal center via its 3-hydroxychromone function. Moreover, enzyme/substrate interactions within the cavity impose steric hindrances to quercetin that may activate a direct dioxygen addition on the substrate. (c) 2008 Wiley-Liss, Inc.

A CRISPR/Cas9 and Cre/Lox system-based express vaccine development strategy against re-emerging Pseudorabies virus.

PubMed

Liang, Xun; Sun, Leqiang; Yu, Teng; Pan, Yongfei; Wang, Dongdong; Hu, Xueying; Fu, Zhenfang; He, Qigai; Cao, Gang

2016-01-18

Virus evolves rapidly to escape vaccine-induced immunity, posing a desperate demand for efficient vaccine development biotechnologies. Here we present an express vaccine development strategy based on CRISPR/Cas9 and Cre/Lox system against re-emerging Pseudorabies virus, which caused the recent devastating swine pseudorabies outbreak in China. By CRISPR/Cas9 system, the virulent genes of the newly isolated strain were simultaneously substituted by marker genes, which were subsequently excised using Cre/Lox system for vaccine safety concern. Notably, single cell FACS technology was applied to further promote virus purification efficiency. The combination of these state-of-art technologies greatly accelerated vaccine development. Finally, vaccination and challenge experiments proved this vaccine candidate's protective efficacy in pigs and the promise to control current pseudorabies outbreak. This is, to our knowledge, the first successful vaccine development based on gene edit technologies, demonstrating these technologies leap from laboratory to industry. It may pave the way for future express antiviral vaccine development.

A CRISPR/Cas9 and Cre/Lox system-based express vaccine development strategy against re-emerging Pseudorabies virus

PubMed Central

Liang, Xun; Sun, Leqiang; Yu, Teng; Pan, Yongfei; Wang, Dongdong; Hu, Xueying; Fu, Zhenfang; He, Qigai; Cao, Gang

2016-01-01

Virus evolves rapidly to escape vaccine-induced immunity, posing a desperate demand for efficient vaccine development biotechnologies. Here we present an express vaccine development strategy based on CRISPR/Cas9 and Cre/Lox system against re-emerging Pseudorabies virus, which caused the recent devastating swine pseudorabies outbreak in China. By CRISPR/Cas9 system, the virulent genes of the newly isolated strain were simultaneously substituted by marker genes, which were subsequently excised using Cre/Lox system for vaccine safety concern. Notably, single cell FACS technology was applied to further promote virus purification efficiency. The combination of these state-of-art technologies greatly accelerated vaccine development. Finally, vaccination and challenge experiments proved this vaccine candidate’s protective efficacy in pigs and the promise to control current pseudorabies outbreak. This is, to our knowledge, the first successful vaccine development based on gene edit technologies, demonstrating these technologies leap from laboratory to industry. It may pave the way for future express antiviral vaccine development. PMID:26777545

Coffee with cinnamon - impact of phytochemicals interactions on antioxidant and anti-inflammatory in vitro activity.

PubMed

Durak, Agata; Gawlik-Dziki, Urszula; Pecio, Lukasz

2014-11-01

This paper evaluates the potential bioaccessibility and interactions between antiradical and anti-inflammatory compounds from coffee and cinnamon. Results obtained for whole plant material extracts were compared with those for chlorogenic and cinnamic acids (the main bioactive constituents of the study material). All samples, coffee, cinnamon and a mixture of the two showed abilities to scavenge free radicals and to inhibit lipoxygenase (LOX) activity. Both activities increased after simulated gastrointestinal digestion. In the mixture antiradical phytochemicals acted antagonistically - isoboles adopted the convex form. The same interactions were determined for chemical standards. The water-extractable LOX inhibitors acted synergistically - the isobole curve was "concave". The same type of interaction was determined for standard compounds. Interestingly, after digestion in vitro a slight antagonism in the action of LOX inhibitors was observed. The results show that the food matrix and/or its changes during digestion may play an important role in creating the biological properties. Copyright © 2014 Elsevier Ltd. All rights reserved.

A Tet-on and Cre-loxP Based Genetic Engineering System for Convenient Recycling of Selection Markers in Penicillium oxalicum

PubMed Central

Jiang, Baojie; Zhang, Ruiqin; Feng, Dan; Wang, Fangzhong; Liu, Kuimei; Jiang, Yi; Niu, Kangle; Yuan, Quanquan; Wang, Mingyu; Wang, Hailong; Zhang, Youming; Fang, Xu

2016-01-01

The lack of selective markers has been a key problem preventing multistep genetic engineering in filamentous fungi, particularly for industrial species such as the lignocellulose degrading Penicillium oxalicum JUA10-1(formerly named as Penicillium decumbens). To resolve this problem, we constructed a genetic manipulation system taking advantage of two established genetic systems: the Cre-loxP system and Tet-on system in P. oxalicum JUA10-1. This system is efficient and convenient. The expression of Cre recombinase was activated by doxycycline since it was controlled by Tet-on system. Using this system, two genes, ligD and bglI, were sequentially disrupted by loxP flanked ptrA. The successful application of this procedure will provide a useful tool for genetic engineering in filamentous fungi. This system will also play an important role in improving the productivity of interesting products and minimizing by-product when fermented by filamentous fungi. PMID:27148179

The performance of a piezoelectric-sensor-based SHM system under a combined cryogenic temperature and vibration environment

NASA Astrophysics Data System (ADS)

Qing, Xinlin P.; Beard, Shawn J.; Kumar, Amrita; Sullivan, Kevin; Aguilar, Robert; Merchant, Munir; Taniguchi, Mike

2008-10-01

A series of tests have been conducted to determine the survivability and functionality of a piezoelectric-sensor-based active structural health monitoring (SHM) SMART Tape system under the operating conditions of typical liquid rocket engines such as cryogenic temperature and vibration loads. The performance of different piezoelectric sensors and a low temperature adhesive under cryogenic temperature was first investigated. The active SHM system for liquid rocket engines was exposed to flight vibration and shock environments on a simulated large booster LOX-H2 engine propellant duct conditioned to cryogenic temperatures to evaluate the physical robustness of the built-in sensor network as well as operational survivability and functionality. Test results demonstrated that the developed SMART Tape system can withstand operational levels of vibration and shock energy on a representative rocket engine duct assembly, and is functional under the combined cryogenic temperature and vibration environment.

Successful nucleofection of rat adipose-derived stroma cells with Ambystoma mexicanum epidermal lipoxygenase (AmbLOXe).

PubMed

Fülbier, Angela; Schnabel, Reinhild; Michael, Stefanie; Vogt, Peter M; Strauß, Sarah; Reimers, Kerstin; Radtke, Christine

2014-10-09

Adipose-derived stroma cells (ASCs) are attractive cells for cell-based gene therapy but are generally difficult to transfect. Nucleofection has proven to be an efficient method for transfection of primary cells. Therefore, we used this technique to transfect ASCs with a vector encoding for Ambystoma mexicanum epidermal lipoxygenase (AmbLOXe) which is a promising bioactive enzyme in regenerative processes. Thereby, we thought to even further increase the large regenerative potential of the ASCs. ASCs were isolated from the inguinal fat pad of Lewis rats and were subsequently transfected in passage 1 using Nucleofector® 2b and the hMSC Nucleofector kit. Transfection efficiency was determined measuring co-transfected green fluorescent protein (GFP) in a flow cytometer and gene expression in transfected cells was detected by reverse transcription polymerase chain reaction (RT-PCR). Moreover, cell migration was assessed using a scratch assay and results were tested for statistical significance with ANOVA followed by Bonferroni's post hoc test. High initial transfection rates were achieved with an average of 79.8 ± 2.82% of GFP positive cells although longer cultivation periods reduced the number of positive cells to below 5% after four passages. Although successful production of AmbLOXe transcript could be proven the gene product had no measureable effect on cell migration. Our study demonstrates the feasibility of ASCs to serve as a vehicle of AmbLOXe transport for gene therapeutic purposes in regenerative medicine. One potential field of applications could be peripheral nerve injuries.

Characterization of a Gene Coding for the Complement System Component FB from Loxosceles laeta Spider Venom Glands.

PubMed

Myamoto, Daniela Tiemi; Pidde-Queiroz, Giselle; Gonçalves-de-Andrade, Rute Maria; Pedroso, Aurélio; van den Berg, Carmen W; Tambourgi, Denise V

2016-01-01

The human complement system is composed of more than 30 proteins and many of these have conserved domains that allow tracing the phylogenetic evolution. The complement system seems to be initiated with the appearance of C3 and factor B (FB), the only components found in some protostomes and cnidarians, suggesting that the alternative pathway is the most ancient. Here, we present the characterization of an arachnid homologue of the human complement component FB from the spider Loxosceles laeta. This homologue, named Lox-FB, was identified from a total RNA L. laeta spider venom gland library and was amplified using RACE-PCR techniques and specific primers. Analysis of the deduced amino acid sequence and the domain structure showed significant similarity to the vertebrate and invertebrate FB/C2 family proteins. Lox-FB has a classical domain organization composed of a control complement protein domain (CCP), a von Willebrand Factor domain (vWFA), and a serine protease domain (SP). The amino acids involved in Mg2+ metal ion dependent adhesion site (MIDAS) found in the vWFA domain in the vertebrate C2/FB proteins are well conserved; however, the classic catalytic triad present in the serine protease domain is not conserved in Lox-FB. Similarity and phylogenetic analyses indicated that Lox-FB shares a major identity (43%) and has a close evolutionary relationship with the third isoform of FB-like protein (FB-3) from the jumping spider Hasarius adansoni belonging to the Family Salcitidae.

Characterization of a Gene Coding for the Complement System Component FB from Loxosceles laeta Spider Venom Glands

PubMed Central

Myamoto, Daniela Tiemi; Pidde-Queiroz, Giselle; Gonçalves-de-Andrade, Rute Maria; Pedroso, Aurélio; van den Berg, Carmen W.; Tambourgi, Denise V.

2016-01-01

The human complement system is composed of more than 30 proteins and many of these have conserved domains that allow tracing the phylogenetic evolution. The complement system seems to be initiated with the appearance of C3 and factor B (FB), the only components found in some protostomes and cnidarians, suggesting that the alternative pathway is the most ancient. Here, we present the characterization of an arachnid homologue of the human complement component FB from the spider Loxosceles laeta. This homologue, named Lox-FB, was identified from a total RNA L. laeta spider venom gland library and was amplified using RACE-PCR techniques and specific primers. Analysis of the deduced amino acid sequence and the domain structure showed significant similarity to the vertebrate and invertebrate FB/C2 family proteins. Lox-FB has a classical domain organization composed of a control complement protein domain (CCP), a von Willebrand Factor domain (vWFA), and a serine protease domain (SP). The amino acids involved in Mg2+ metal ion dependent adhesion site (MIDAS) found in the vWFA domain in the vertebrate C2/FB proteins are well conserved; however, the classic catalytic triad present in the serine protease domain is not conserved in Lox-FB. Similarity and phylogenetic analyses indicated that Lox-FB shares a major identity (43%) and has a close evolutionary relationship with the third isoform of FB-like protein (FB-3) from the jumping spider Hasarius adansoni belonging to the Family Salcitidae. PMID:26771533

Modulation of Cox-1, 5-, 12- and 15-Lox by Popular Herbal Remedies Used in Southern Italy Against Psoriasis and Other Skin Diseases

PubMed Central

Bader, Ammar; Martini, Francesca; Schinella, Guillermo R; Rios, Jose L; Prieto, Jose M

2015-01-01

Acanthus mollis (Acanthaceae), Achillea ligustica, Artemisia arborescens and Inula viscosa (Asteraceae) are used in Southern Italy against psoriasis and other skin diseases that occur with an imbalanced production of eicosanoids. We here assessed their in vitro effects upon 5-, 12-, 15-LOX and COX-1 enzymes as well as NFκB activation in intact cells as their possible therapeutic targets. All methanol crude extracts inhibited both 5-LOX and COX-1 activities under 200 µg/mL, without significant effects on the 12-LOX pathway or any relevant in vitro free radical scavenging activity. NFκB activation was prevented by all extracts but A. mollis. Interestingly, A. ligustica, A. arborescens and A. mollis increased the biosynthesis of 15(S)-HETE, an anti-inflammatory eicosanoid. A. ligustica (IC50 = 49.5 µg/mL) was superior to Silybum marianum (IC50 = 147.8 µg/mL), which we used as antipsoriatic herbal medicine of reference. Its n-hexane, dichloromethane and ethyl acetate fractions had also inhibitory effects on the LTB4 biosynthesis (IC50s = 9.6, 20.3 and 68 µg/mL, respectively) evidencing that the apolar extracts of A. ligustica are promising active herbal ingredients for future phytotherapeutical products targeting psoriasis. © 2014 The Authors. Phytotherapy Research published by John Wiley & Sons Ltd. PMID:25278440

Dimethyl ester of bilirubin exhibits anti-inflammatory activity through inhibition of secretory phospholipase A2, lipoxygenase and cyclooxygenase.

PubMed

Joshi, Vikram; Umashankara, M; Ramakrishnan, Chandrasekaran; Nanjaraj Urs, Ankanahalli N; Suvilesh, Kanve Nagaraj; Velmurugan, Devadasan; Rangappa, Kanchugarakoppal S; Vishwanath, Bannikuppe Sannanaik

2016-05-15

Overproduction of arachidonic acid (AA) mediated by secretory phospholipase A2 group IIA (sPLA2IIA) is a hallmark of many inflammatory disorders. AA is subsequently converted into pro-inflammatory eicosanoids through 5-lipoxygenase (5-LOX) and cyclooxygenase-1/2 (COX-1/2) activities. Hence, inhibition of sPLA2IIA, 5-LOX and COX-1/2 activities is critical in regulating inflammation. We have previously reported unconjugated bilirubin (UCB), an endogenous antioxidant, as sPLA2IIA inhibitor. However, lipophilic UCB gets conjugated in liver with glucuronic acid into hydrophilic conjugated bilirubin (CB). Since hydrophobicity is pre-requisite for sPLA2IIA inhibition, conjugation reduces the efficacy of UCB. In this regard, UCB was chemically modified and derivatives were evaluated for sPLA2IIA, 5-LOX and COX-1/2 inhibition. Among the derivatives, BD1 (dimethyl ester of bilirubin) exhibited ∼ 3 fold greater inhibitory potency towards sPLA2IIA compared to UCB. Both UCB and BD1 inhibited human 5-LOX and COX-2 activities; however only BD1 inhibited AA induced platelet aggregation. Molecular docking studies demonstrated BD1 as better inhibitor of aforesaid enzymes than UCB and other endogenous antioxidants. These data suggest that BD1 exhibits strong anti-inflammatory activity through inhibition of AA cascade enzymes which is of great therapeutic importance. Copyright © 2016 Elsevier Inc. All rights reserved.

Synergistic anti-tumor effects of melatonin and PUFAs from walnuts in a murine mammary adenocarcinoma model.

PubMed

Garcia, Carolina P; Lamarque, Alicia L; Comba, Andrea; Berra, María A; Silva, Renata A; Labuckas, Diana O; Das, Undurti N; Eynard, Aldo R; Pasqualini, Maria E

2015-04-01

The aim of this study was to determine the effects of some polyunsaturated fatty acids plus phytomelatonin from walnuts in the development of mammary gland adenocarcinoma. BALB/c mice were fed a semisynthetic diet supplemented with either 6% walnut oil and 8% walnut flour containing phytomelatonin (walnut diet: WD); or 6% corn oil plus commercial melatonin (melatonin diet: MD), or the control group (CD), which received only 6% of corn oil. Membrane fatty acids of tumor cells (TCs) were analyzed by gas liquid chromatography, cyclooxygenase (COX) and lipoxygenase (LOX) derivatives, and plasma melatonin by high-performance liquid chromatography; apoptosis and tumor-infiltrating lymphocytes by flow cytometry. TCs from the MD and WD mice showed significant decreases in linoleic acid compared with the CD group (P < 0.05). Significantly lower levels of LOX-[13(S)-HODE] were found in TCs from the MD and WD group than in CD (P < 0.0001). COX-[12(S)-HHT] was lower and 12 LOX-[12(S)-HETE] was higher in TCs from the MD group than form the WD and CD arms (P < 0.05). Plasma melatonin, apoptosis, tumor infiltration, and survival time were significantly lower in CD mice than in MD and WD mice (P < 0.05). This study shows that melatonin, along with polyunsaturated fatty acids, exerts a selective inhibition of some COX and LOX activities and has a synergistic anti-tumor effect on a mammary gland adenocarcinoma model. Published by Elsevier Inc.

A single active site residue directs oxygenation stereospecificity in lipoxygenases: Stereocontrol is linked to the position of oxygenation

PubMed Central

Coffa, Gianguido; Brash, Alan R.

2004-01-01

Lipoxygenases are a class of dioxygenases that form hydroperoxy fatty acids with distinct positional and stereo configurations. Several amino acid residues influencing regiospecificity have been identified, whereas the basis of stereocontrol is not understood. We have now identified a single residue in the lipoxygenase catalytic domain that is important for stereocontrol; it is conserved as an Ala in S lipoxygenases and a Gly in R lipoxygenases. Our results with mutation of the conserved Ala to Gly in two S lipoxygenases (mouse 8S-LOX and human 15-LOX-2) and the corresponding Gly–Ala substitution in two R lipoxygenases (human 12R-LOX and coral 8R-LOX) reveal that the basis for R or S stereo-control also involves a switch in the position of oxygenation on the substrate. After the initial hydrogen abstraction, antarafacial oxygenation at one end or the other of the activated pair of double bonds (pentadiene) gives, for example, 8S or 12R product. The Ala residue promotes oxygenation on the reactive pentadiene at the end deep in the substrate binding pocket and S stereochemistry of the product hydroperoxide, and a Gly residue promotes oxygenation at the proximal end of the reactive pentadiene resulting in R stereochemistry. A model of lipoxygenase reaction specificity is proposed in which product regiochemistry and stereochemistry are determined by fixed relationships between substrate orientation, hydrogen abstraction, and the Gly or Ala residue we have identified. PMID:15496467

Primary Coenzyme Q Deficiency in Pdss2 Mutant Mice Causes Isolated Renal Disease

PubMed Central

Haase, Volker H.; King, Rhonda; Polyak, Erzsebet; Selak, Mary; Yudkoff, Marc; Hancock, Wayne W.; Meade, Ray; Saiki, Ryoichi; Lunceford, Adam L.; Clarke, Catherine F.; Gasser, David L.

2008-01-01

Coenzyme Q (CoQ) is an essential electron carrier in the respiratory chain whose deficiency has been implicated in a wide variety of human mitochondrial disease manifestations. Its multi-step biosynthesis involves production of polyisoprenoid diphosphate in a reaction that requires the enzymes be encoded by PDSS1 and PDSS2. Homozygous mutations in either of these genes, in humans, lead to severe neuromuscular disease, with nephrotic syndrome seen in PDSS2 deficiency. We now show that a presumed autoimmune kidney disease in mice with the missense Pdss2kd/kd genotype can be attributed to a mitochondrial CoQ biosynthetic defect. Levels of CoQ9 and CoQ10 in kidney homogenates from B6.Pdss2kd/kd mutants were significantly lower than those in B6 control mice. Disease manifestations originate specifically in glomerular podocytes, as renal disease is seen in Podocin/cre,Pdss2loxP/loxP knockout mice but not in conditional knockouts targeted to renal tubular epithelium, monocytes, or hepatocytes. Liver-conditional B6.Alb/cre,Pdss2loxP/loxP knockout mice have no overt disease despite demonstration that their livers have undetectable CoQ9 levels, impaired respiratory capacity, and significantly altered intermediary metabolism as evidenced by transcriptional profiling and amino acid quantitation. These data suggest that disease manifestations of CoQ deficiency relate to tissue-specific respiratory capacity thresholds, with glomerular podocytes displaying the greatest sensitivity to Pdss2 impairment. PMID:18437205

Selectable antibiotic resistance marker gene-free transgenic rice harbouring the garlic leaf lectin gene exhibits resistance to sap-sucking planthoppers.

PubMed

Sengupta, Subhadipa; Chakraborti, Dipankar; Mondal, Hossain A; Das, Sampa

2010-03-01

Rice, the major food crop of world is severely affected by homopteran sucking pests. We introduced coding sequence of Allium sativum leaf agglutinin, ASAL, in rice cultivar IR64 to develop sustainable resistance against sap-sucking planthoppers as well as eliminated the selectable antibiotic-resistant marker gene hygromycin phosphotransferase (hpt) exploiting cre/lox site-specific recombination system. An expression vector was constructed containing the coding sequence of ASAL, a potent controlling agent against green leafhoppers (GLH, Nephotettix virescens) and brown planthopper (BPH, Nilaparvata lugens). The selectable marker (hpt) gene cassette was cloned within two lox sites of the same vector. Alongside, another vector was developed with chimeric cre recombinase gene cassette. Reciprocal crosses were performed between three single-copy T(0) plants with ASAL- lox-hpt-lox T-DNA and three single-copy T(0) plants with cre-bar T-DNA. Marker gene excisions were detected in T(1) hybrids through hygromycin sensitivity assay. Molecular analysis of T(1) plants exhibited 27.4% recombination efficiency. T(2) progenies of L03C04(1) hybrid parent showed 25% cre negative ASAL-expressing plants. Northern blot, western blot and ELISA showed significant level of ASAL expression in five marker-free T(2) progeny plants. In planta bioassay of GLH and BPH performed on these T(2) progenies exhibited radical reduction in survivability and fecundity compared with the untransformed control plants.

TALEN-Based Mutagenesis of Lipoxygenase LOX3 Enhances the Storage Tolerance of Rice (Oryza sativa) Seeds.

PubMed

Ma, Lei; Zhu, Fugui; Li, Zhenwei; Zhang, Jianfu; Li, Xin; Dong, Jiangli; Wang, Tao

2015-01-01

The deterioration of rice grain reduces the quality of rice, resulting in serious economic losses for farmers. Lipoxygenases (LOXs) catalyze the dioxygenation of polyunsaturated fatty acids with at least one cis,cis-1,4-pentadiene to form hydroperoxide, which is a major factor influencing seed longevity and viability. Recently, genome editing, an essential tool employed in reverse genetics, has been used experimentally to investigate basic plant biology or to modify crop plants for the improvement of important agricultural traits. In this study, we performed targeted mutagenesis in rice using transcription activator-like effector nucleases (TALENs) to improve seed storability. A modified ligation-independent cloning method (LIC) was employed to allow for the quick and efficient directional insertion of TALEN monomer modules into destination vectors used in plants. We demonstrated the feasibility and flexibility of the technology by developing a set of modular vectors for genome editing. After construction and validation, the TALEN pairs were used to create stable transgenic rice lines via Agrobacterium-mediated transformation. One heterozygous mutant (4%) was recovered from 25 transgenic NPTII-resistant lines, and the mutation was transmitted to the next generation. Further molecular and protein level experiments verified LOX3 deficiency and demonstrated the improvement of seed storability. Our work provides a flexible genome editing tool for improving important agronomic traits, as well as direct evidence that Lox3 has only a limited impact on seed longevity.

TALEN-Based Mutagenesis of Lipoxygenase LOX3 Enhances the Storage Tolerance of Rice (Oryza sativa) Seeds

PubMed Central

Li, Zhenwei; Zhang, Jianfu; Li, Xin; Dong, Jiangli; Wang, Tao

2015-01-01

The deterioration of rice grain reduces the quality of rice, resulting in serious economic losses for farmers. Lipoxygenases (LOXs) catalyze the dioxygenation of polyunsaturated fatty acids with at least one cis,cis-1,4-pentadiene to form hydroperoxide, which is a major factor influencing seed longevity and viability. Recently, genome editing, an essential tool employed in reverse genetics, has been used experimentally to investigate basic plant biology or to modify crop plants for the improvement of important agricultural traits. In this study, we performed targeted mutagenesis in rice using transcription activator-like effector nucleases (TALENs) to improve seed storability. A modified ligation-independent cloning method (LIC) was employed to allow for the quick and efficient directional insertion of TALEN monomer modules into destination vectors used in plants. We demonstrated the feasibility and flexibility of the technology by developing a set of modular vectors for genome editing. After construction and validation, the TALEN pairs were used to create stable transgenic rice lines via Agrobacterium-mediated transformation. One heterozygous mutant (4%) was recovered from 25 transgenic NPTII-resistant lines, and the mutation was transmitted to the next generation. Further molecular and protein level experiments verified LOX3 deficiency and demonstrated the improvement of seed storability. Our work provides a flexible genome editing tool for improving important agronomic traits, as well as direct evidence that Lox3 has only a limited impact on seed longevity. PMID:26641666

6-Shogaol Protects against Oxidized LDL-Induced Endothelial Injruries by Inhibiting Oxidized LDL-Evoked LOX-1 Signaling

PubMed Central

Wang, Yun kai; Hong, Ya Jun; Yao, Yong hua; Huang, Xiao Min; Liu, Xue Bo; Zhang, Chun Yu; Zhang, Lei; Xu, Xiaoliang Leon

2013-01-01

Endothelial dysfunction and oxLDL are believed to be early and critical events in atherogenesis. 6-Shogaol is the major bioactive compound present in Zingiber officinale and possesses the anti-atherosclerotic effect. However, the mechanisms remain poorly understood. The goal of this study was to investigate the effects of 6-shogaol on oxLDL-induced Human umbilical vein endothelial cells (HUVECs) injuries and its possible molecular mechanisms. Hence, we studied the effects of 6-shogaol on cell apoptosis, cellular reactive oxygen species (ROS), NF-κB activation, Bcl-2 expression, and caspase -3, -8, -9 activities. In addition, E-selectin, MCP-1, and ICAM-1 were determined by ELISA. Our study show that oxLDL increased LOX-1 expression, ROS levels, NF-κB, caspases-9 and -3 activation and decreased Bcl-2 expression in HUVECs. These alterations were attenuated by 6-shogaol. Cotreatment with 6-shogaol and siRNA of LOX-1 synergistically reduced oxLDL-induced caspases -9, -3 activities and cell apoptosis. Overexpression of LOX-1 attenuated the protection by 6-shogaol and suppressed the effects of 6-shogaol on oxLDL-induced oxidative stress. In addition, oxLDL enhanced the activation of NF-κB and expression of adhesion molecules. Pretreatment with 6-shogaol, however, exerted significant cytoprotective effects in all events. Our data indicate that 6-shogaol might be a potential natural antiapoptotic agent for the treatment of atherosclerosis. PMID:23533490

6-Shogaol Protects against Oxidized LDL-Induced Endothelial Injruries by Inhibiting Oxidized LDL-Evoked LOX-1 Signaling.

PubMed

Wang, Yun Kai; Hong, Ya Jun; Yao, Yong Hua; Huang, Xiao Min; Liu, Xue Bo; Zhang, Chun Yu; Zhang, Lei; Xu, Xiaoliang Leon

2013-01-01

Endothelial dysfunction and oxLDL are believed to be early and critical events in atherogenesis. 6-Shogaol is the major bioactive compound present in Zingiber officinale and possesses the anti-atherosclerotic effect. However, the mechanisms remain poorly understood. The goal of this study was to investigate the effects of 6-shogaol on oxLDL-induced Human umbilical vein endothelial cells (HUVECs) injuries and its possible molecular mechanisms. Hence, we studied the effects of 6-shogaol on cell apoptosis, cellular reactive oxygen species (ROS), NF- κ B activation, Bcl-2 expression, and caspase -3, -8, -9 activities. In addition, E-selectin, MCP-1, and ICAM-1 were determined by ELISA. Our study show that oxLDL increased LOX-1 expression, ROS levels, NF- κ B, caspases-9 and -3 activation and decreased Bcl-2 expression in HUVECs. These alterations were attenuated by 6-shogaol. Cotreatment with 6-shogaol and siRNA of LOX-1 synergistically reduced oxLDL-induced caspases -9, -3 activities and cell apoptosis. Overexpression of LOX-1 attenuated the protection by 6-shogaol and suppressed the effects of 6-shogaol on oxLDL-induced oxidative stress. In addition, oxLDL enhanced the activation of NF- κ B and expression of adhesion molecules. Pretreatment with 6-shogaol, however, exerted significant cytoprotective effects in all events. Our data indicate that 6-shogaol might be a potential natural antiapoptotic agent for the treatment of atherosclerosis.

Combustion Stability Characteristics of the Project Morpheus Liquid Oxygen/Liquid Methane Main Engine

NASA Technical Reports Server (NTRS)

Melcher, J. C.; Morehead, Robert L.

2014-01-01

The Project Morpheus liquid oxygen (LOX) / liquid methane rocket engines demonstrated acousticcoupled combustion instabilities during sea-level ground-based testing at the NASA Johnson Space Center (JSC) and Stennis Space Center (SSC). High-amplitude, 1T, 1R, 1T1R (and higher order) modes appear to be triggered by injector conditions. The instability occurred during the Morpheus-specific engine ignition/start sequence, and did demonstrate the capability to propagate into mainstage. However, the instability was never observed to initiate during mainstage, even at low power levels. The Morpheus main engine is a JSC-designed 5,000 lbf-thrust, 4:1 throttling, pressure-fed cryogenic engine using an impinging element injector design. Two different engine designs, named HD4 and HD5, and two different builds of the HD4 engine all demonstrated similar instability characteristics. Through the analysis of more than 200 hot fire tests on the Morpheus vehicle and SSC test stand, a relationship between ignition stability and injector/chamber pressure was developed. The instability has the distinct characteristic of initiating at high relative injection pressure drop (dP) at low chamber pressure (Pc); i.e., instabilities initiated at high dP/Pc at low Pc during the start sequence. The high dP/Pc during start results during the injector /chamber chill-in, and is enhanced by hydraulic flip in the injector orifice elements. Because of the fixed mixture ratio of the existing engine design (the main valves share a common actuator), it is not currently possible to determine if LOX or methane injector dP/Pc were individual contributors (i.e., LOX and methane dP/Pc typically trend in the same direction within a given test). The instability demonstrated initiation characteristic of starting at or shortly after methane injector chillin. Colder methane (e.g., sub-cooled) at the injector inlet prior to engine start was much more likely to result in an instability. A secondary effect of LOX sub-cooling was also possibly observed; greater LOX sub- cooling improved stability. Some tests demonstrated a low-amplitude 1L-1T instability prior to LOX injector chill-in. The Morpheus main engine also demonstrated chug instabilities during some engine shutdown sequences on the flight vehicle and SSC test stand. The chug instability was also infrequently observed during the startup sequence. The chug instabilities predictably initiated at low dP/Pc at low Pc. The chug instabilities were always self-limiting; startup chug instabilities terminated during throttle-up and shutdown chug instabilities decayed by shutdown termination.

Insight into durum wheat Lpx-B1: a small gene family coding for the lipoxygenase responsible for carotenoid bleaching in mature grains.

PubMed

Verlotta, Angelo; De Simone, Vanessa; Mastrangelo, Anna M; Cattivelli, Luigi; Papa, Roberto; Trono, Daniela

2010-11-26

The yellow colour of pasta products is one of the main criteria used by consumers to assess pasta quality. This character is due to the presence of carotenoid pigments in semolina. During pasta processing, oxidative degradation of carotenoid pigments occurs mainly due to lipoxygenase (LOX). In durum wheat (Triticum durum Desf.), two Lpx-1 genes have been identified on chromosome 4B, Lpx-B1.1 and Lpx-B1.2, and evidences have been reported that the deletion of Lpx-B1.1 is associated with a strong reduction in LOX activity in semolina. In the present study, we characterised the Lpx-B1 gene family identified in a durum wheat germplasm collection and related the distribution and expression of the Lpx-B1 genes and alleles to variations in LOX activity in the mature grains. In addition to the already known Lpx-B1.1 and Lpx-B1.2 genes, a new gene was identified, Lpx-B1.3, along with three different Lpx-B1.1 alleles, Lpx-B1.1a, Lpx-B1.1b and the partially deleted Lpx-B1.1c. Screening of the germplasm collection showed that all of the genotypes have one of the three Lpx-B1.1 alleles, associated with either Lpx-B1.2 or Lpx-B1.3, thus showing that in this collection the two genes are alternatives. Therefore, based on Lpx-B1 distribution, three different haplotypes were distinguished: haplotype I, carrying Lpx-B1.3 and the Lpx-B1.1b allele; haplotype II carrying Lpx-B1.2 and the Lpx-B1.1a allele; and haplotype III carrying Lpx-B1.2 and the Lpx-B1.1c allele. Determination of Lpx-B1 transcript abundance and total LOX activity in mature grains revealed differences among these three haplotypes: haplotypes I, II and III showed high, intermediate and low levels, respectively, of functional Lpx-B1 transcripts and enzymatic activity. In this germplasm collection, the Lpx-B1 gene family accounts for most of the total LOX activity in the mature grains. Information on these Lpx-B1 haplotypes provides significant improvement for prediction of LOX-1 activity levels in mature grains, and will therefore help in breeding programmes aimed at selection of new durum wheat genotypes with higher carotenoid contents in their end products.

Electronegative Low-Density Lipoprotein Increases C-Reactive Protein Expression in Vascular Endothelial Cells through the LOX-1 Receptor

PubMed Central

Chu, Chih-Sheng; Wang, Yu-Chen; Lu, Long-Sheng; Walton, Brian; Yilmaz, H. Ramazan; Huang, Roger Y.; Sawamura, Tatsuya; Dixon, Richard A. F.; Lai, Wen-Ter; Chen, Chu-Huang; Lu, Jonathan

2013-01-01

Objectives Increased plasma C-reactive protein (CRP) levels are associated with the occurrence and severity of acute coronary syndrome. We investigated whether CRP can be generated in vascular endothelial cells (ECs) after exposure to the most electronegative subfraction of low-density lipoprotein (LDL), L5, which is atherogenic to ECs. Because L5 and CRP are both ligands for the lectin-like oxidized LDL receptor-1 (LOX-1), we also examined the role of LOX-1. Methods and Results Plasma LDL samples isolated from asymptomatic hypercholesterolemic (LDL cholesterol [LDL-C] levels, 154.6±20 mg/dL; n = 7) patients and normocholesterolemic (LDL-C levels, 86.1±21 mg/dL; P<0.001; n = 7) control individuals were chromatographically resolved into 5 subfractions, L1-L5. The L5 percentage (L5%) and the plasma L5 concentration ([L5]  =  L5% × LDL-C) in the patient and control groups were 8.1±2% vs. 2.3±1% (P<0.001) and 12.6±4 mg/dL vs. 1.9±1 mg/dL (P<0.001), respectively. In hypercholesterolemic patients treated with atorvastatin for 6 months (10 mg/day), [L5] decreased from 12.6±4 mg/dL to 4.5±1.1 mg/dL (P = 0.011; n = 5), whereas both [L5] and L5% returned to baseline levels in 2 noncompliant patients 3 months after discontinuation. In cultured human aortic ECs (HAECs), L5 upregulated CRP expression in a dose- and time-dependent manner up to 2.5-fold (P<0.01), whereas the least electronegative subfraction, L1, had no effect. DiI-labeled L1, internalized through the LDL receptor, became visible inside HAECs within 30 seconds. In contrast, DiI-labeled L5, internalized through LOX-1, became apparent after 5 minutes. L5-induced CRP expression manifested at 30 minutes and was attenuated by neutralizing LOX-1. After 30 minutes, L5 but not L1 induced reactive oxygen species (ROS) production. Both L5-induced ROS and CRP production were attenuated by ROS inhibitor N-acetyl cysteine. Conclusions Our results suggest that CRP, L5, and LOX-1 form a cyclic mechanism in atherogenesis and that reducing plasma L5 levels with atorvastatin disrupts the vascular toxicity of L5. PMID:23950953

Miniature Joule Thomson (JT) CryoCoolers for Propellant Management

NASA Technical Reports Server (NTRS)

Kapat, Jay; Chow, Louis

2002-01-01

A proof-of-concept project is proposed here that would attempt to demonstrate how miniature cryocoolers can be used to chill the vacuum jacket line of a propellant transfer line and thus to achieve transfer line pre-chill, zero boil off and possible propellant densification. The project would be performed both at UCF and KSC, with all of the cryogenic testing taking place in the KSC cryogenic test bed. A LN2 line available in that KSC test facility would serve to simulate a LOX transfer line. Under this project, miniature and highly efficient cold heads would be designed. Two identical cold heads will be fabricated and then integrated with a JT-type cryogenic system (consisting of a common compressor and a common external heat exchanger). The two cold heads will be integrated into the vacuum jacket of a LN2 line in the KSC cryo lab, where the testing will take place.

Test Results for a Non-toxic, Dual Thrust Reaction Control Engine

NASA Technical Reports Server (NTRS)

Robinson, Philip J.; Veith, Eric M.; Turpin, Alicia A.

2005-01-01

A non-toxic, dual thrust reaction control engine (RCE) was successfully tested over a broad range of operating conditions at the Aerojet Sacramento facility. The RCE utilized LOX/Ethanol propellants; and was tested in steady state and pulsing modes at 25-lbf thrust (vernier) and at 870-lbf thrust (primary). Steady state vernier tests vaned chamber pressure (Pc) from 0.78 to 5.96 psia, and mixture ratio (MR) from 0.73 to 1.82, while primary steady state tests vaned Pc from 103 to 179 psia and MR from 1.33 to 1.76. Pulsing tests explored EPW from 0.080 to 10 seconds and DC from 5 to 50 percent at both thrust levels. Vernier testing accumulated a total of 6,670 seconds of firing time, and 7,215 pulses, and primary testing accumulated a total of 2,060 seconds of firing time and 3,646 pulses.

United States Air Force Wipe Solvent Testing

NASA Technical Reports Server (NTRS)

Hornung, Steven D.; Beeson, Harold D.

2000-01-01

The Wright-Patterson Air Force Base (WPAFB), as part of the Air Force Material Command, requested that NASA Johnson Space Center (JSC) White Sands Test Facility (WSTF) conduct testing and analyses in support of the United States Air Force Wipe Solvent Development Project. The purpose of the wipe solvent project is to develop an alternative to be used by Air Force flight line and maintenance personnel for the wipe cleaning of oxygen equipment. This report provides material compatibility, liquid oxygen (LOX) mechanical impact, autogenous ignition temperature (AIT), and gauge cleaning test data for some of the currently available solvents that may be used to replace CFC-113 and methyl chloroform. It provides data from previous WSTF test programs sponsored by the Naval Sea Systems Command, the Kennedy Space Center, and other NASA programs for the purpose of assisting WP AFB in identifying the best alternative solvents for validation testing.

77 FR 24646 - Open Access and Priority Rights on Interconnection Facilities

Federal Register 2010, 2011, 2012, 2013, 2014

2012-04-25

... multiple generation facilities to transmit power from the generation facility to the integrated... power flows toward the network grid, with no electrical loads between the generation facilities and the... generator expansion plans with milestones for construction of generation facilities and can demonstrate that...

Worldwide Space Launch Vehicles and Their Mainstage Liquid Rocket Propulsion

NASA Technical Reports Server (NTRS)

Rahman, Shamim A.

2010-01-01

Space launch vehicle begins with a basic propulsion stage, and serves as a missile or small launch vehicle; many are traceable to the 1945 German A-4. Increasing stage size, and increasingly energetic propulsion allows for heavier payloads and greater. Earth to Orbit lift capability. Liquid rocket propulsion began with use of storable (UDMH/N2O4) and evolved to high performing cryogenics (LOX/RP, and LOX/LH). Growth versions of SLV's rely on strap-on propulsive stages of either solid propellants or liquid propellants.

Propellant Management and Conditioning within the X-34 Main Propulsion System

NASA Technical Reports Server (NTRS)

Brown, T. M.; McDonald, J. P.; Hedayat, A.; Knight, K. C.; Champion, R. H., Jr.

1998-01-01

The X-34 hypersonic flight vehicle is currently under development by Orbital Sciences Corporation (Orbital). The Main Propulsion ystem as been designed around the liquid propellant Fastrac rocket engine currently under development at NASA Marshall Space Flight Center. This paper presents analyses of the MPS subsystems used to manage the liquid propellants. These subsystems include the propellant tanks, the tank vent/relief subsystem, and the dump/fill/drain subsystem. Analyses include LOX tank chill and fill time estimates, LOX boil-off estimates, propellant conditioning simulations, and transient propellant dump simulations.

Pressure fed thrust chamber technology program

NASA Technical Reports Server (NTRS)

Dunn, Glen M.

1992-01-01

This is the final report for the Pressure Fed Technology Program. It details the design, fabrication, and testing of subscale hardware which successfully characterized Liquid Oxygen Rocket Propulsion (LOX/RP) combustion for low cost pressure fed design. The innovative modular injector design is described in detail as well as hot-fire test results which showed excellent performance. The program summary identifies critical LOX/RP design issues that have been resolved in this testing, and details the low risk development requirements for low cost engines for future Expandable Launch Vehicles (ELV).